Strategies to enhance the response of liver cancer to pharmacological treatments.

In contrast to other types of cancers, there is no available efficient pharmacological treatment to improve the outcomes of patients suffering from major primary liver cancers, i.e., hepatocellular carcinoma and cholangiocarcinoma. This dismal situation is partly due to the existence in these tumors of many different and synergistic mechanisms of resistance, accounting for the lack of response of these patients, not only to classical chemotherapy but also to more modern pharmacological agents based on the inhibition of tyrosine kinase receptors (TKIs) and the stimulation of the immune response against the tumor using immune checkpoint inhibitors (ICIs). This review summarizes the efforts to develop strategies to overcome this severe limitation, including searching for novel drugs derived from synthetic, semisynthetic, or natural products with vectorial properties against therapeutic targets to increase drug uptake or reduce drug export from cancer cells. Besides, immunotherapy is a promising line of research that is already starting to be implemented in clinical practice. Although less successful than in other cancers, the foreseen future for this strategy in treating liver cancers is considerable. Similarly, the pharmacological inhibition of epigenetic targets is highly promising. Many novel "epidrugs," able to act on "writer," "reader," and "eraser" epigenetic players, are currently being evaluated in preclinical and clinical studies. Finally, gene therapy is a broad field of research in the fight against liver cancer chemoresistance, based on the impressive advances recently achieved in gene manipulation. In sum, although the present is still dismal, there is reason for hope in the non-too-distant future.

Synthesis, Characterization, and Potential Usefulness in Liver Function Assessment of Novel Bile Acid Derivatives with Near-Infrared Fluorescence (NIRBAD).

Conventional serum markers often fail to accurately detect cholestasis accompanying many liver diseases. Although elevation in serum bile acid (BA) levels sensitively reflects impaired hepatobiliary function, other factors altering BA pool size and enterohepatic circulation can affect these levels. To develop fluorescent probes for extracorporeal noninvasive hepatobiliary function assessment by real-time monitoring methods, 1,3-dipolar cycloaddition reactions were used to conjugate near-infrared (NIR) fluorochromes with azide-functionalized BA derivatives (BAD). The resulting compounds (NIRBADs) were chromatographically (FC and PTLC) purified (>95%) and characterized by fluorimetry, 1H NMR, and HRMS using ESI ionization coupled to quadrupole TOF mass analysis. Transport studies using CHO cells stably expressing the BA carrier NTCP were performed by flow cytometry. Extracorporeal fluorescence was detected in anesthetized rats by high-resolution imaging analysis. Three NIRBADs were synthesized by conjugating alkynocyanine 718 with cholic acid (CA) at the COOH group via an ester (NIRBAD-1) or amide (NIRBAD-3) spacer, or at the 3α-position by a triazole link (NIRBAD-2). NIRBADs were efficiently taken up by cells expressing NTCP, which was inhibited by taurocholic acid (TCA). Following i.v. administration of NIRBAD-3 to rats, liver uptake and consequent release of NIR fluorescence could be extracorporeally monitored. This transient organ-specific handling contrasted with the absence of release to the intestine of alkynocyanine 718 and the lack of hepatotropism observed with other probes, such as indocyanine green. NIRBAD-3 administration did not alter serum biomarkers of hepatic and renal toxicity. NIRBADs can serve as probes to evaluate hepatobiliary function by noninvasive extracorporeal methods.

Mechanisms of Pharmacoresistance in Hepatocellular Carcinoma: New Drugs but Old Problems.

Hepatocellular carcinoma (HCC) is a malignancy with poor prognosis when diagnosed at advanced stages in which curative treatments are no longer applicable. A small group of these patients may still benefit from transarterial chemoembolization. The only therapeutic option for most patients with advanced HCC is systemic pharmacological treatments based on tyrosine kinase inhibitors (TKIs) and immunotherapy. Available drugs only slightly increase survival, as tumor cells possess additive and synergistic mechanisms of pharmacoresistance (MPRs) prior to or enhanced during treatment. Understanding the molecular basis of MPRs is crucial to elucidate the genetic signature underlying HCC resistome. This will permit the selection of biomarkers to predict drug treatment response and identify tumor weaknesses in a personalized and dynamic way. In this article, we have reviewed the role of MPRs in current first-line drugs and the combinations of immunotherapeutic agents with novel TKIs being tested in the treatment of advanced HCC.

Psychological and Functional Impact of COVID-19 in Long-Term Care Facilities: The COVID-A Study.

OBJECTIVE: To analyze the psychological and functional sequelae of the COVID-19 pandemic among older adults living in long term care facilities (LTCFs). DESIGN: Cohort longitudinal study SETTING ANT PARTICIPANTS: A total of 215 residents ≥ 65 years without moderate-to-severe cognitive impairment, living in five LTCFs in Albacete (Spain). MEASUREMENTS: Baseline on-site data were collected between March - June 2020 and three-month follow-up between June to September 2020. Symptoms of depression, anxiety, posttraumatic stress disorder (PTSD), and sleep disturbances were measured as psychological variables. Disability in basic activities of daily living (BADL), ambulation and frailty were assessed as functional variables. Differences were analyzed in relation to level of comorbidity and test positivity for COVID-19. RESULTS: At baseline, residents with COVID-19 presented worse functionality, higher frailty levels and malnutrition risk compared to non-COVID-19 residents. At three-month follow-up, higher rates of clinically significant depressive symptoms (57.7%), anxiety symptoms (29.3%), PTSD symptoms (19.1%) and sleep disturbances (93.0%) were found among residents regardless of COVID status. Thus, among 215 residents, 101 (47%) experienced a decline in BADL from baseline to the 3-month follow-up (median functional loss = 5 points in Barthel Index). In multivariate analyses, COVID-19 status did not explain either the functional or the ambulation loss. By contrast, residents with low comorbidity and COVID-19 presented higher PTSD symptoms (effect 2.58; 95% CI 0.93 to 4.23) and anxiety symptoms (effect 2.10; 95% CI 0.48 to 3.73) compared to the low comorbidity/non-COVID19 group. CONCLUSION: COVID-19 pandemic was associated, after three-months, with high psychological impact in older adults in LTCFs., specifically with higher post-traumatic stress and anxiety symptoms. Functional decline did not differ in relation to COVID-19 status but could be related to isolation strategies used for pandemic control.

Probability of high-risk genetic matching with oocyte and semen donors: complete gene analysis or genotyping test?

PURPOSE: To estimate the probability of high-risk genetic matching when assisted reproductive techniques (ART) are applied with double gamete donation, following an NGS carrier test based on a complete study of the genes concerned. We then determine the results that would have been obtained if the genotyping tests most widely used in Spanish gamete banks had been applied. METHODS: In this descriptive observational study, 1818 gamete donors were characterised by NGS. The pathogenic variants detected were analysed to estimate the probability of high-risk genetic matching and to determine the results that would have been obtained if the three most commonly used genotyping tests in ART had been applied. RESULTS: The probability of high-risk genetic matching with gamete donation, screened by NGS and complete gene analysis, was 5.5%, versus the 0.6-2.7% that would have been obtained with the genotyping test. A total of 1741 variants were detected, including 607 different variants, of which only 22.6% would have been detected by all three genotyping tests considered and 44.7% of which would not have been detected by any of these tests. CONCLUSION: Our study highlights the considerable heterogeneity of the genotyping tests, which present significant differences in their ability to detect pathogenic variants. The complete study of the genes by NGS considerably reduces reproductive risks when genetic matching is performed with gamete donors. Accordingly, we recommend that carrier screening in gamete donors be carried out using NGS and a complete study with nontargeted analysis of the variants of the screened genes.

Frailty, depression risk and 10-year hospitalization in older adults. The FRADEA study.

OBJECTIVES: To determine whether the interaction between frailty status and depression risk is associated with hospitalization density in older adults. METHODS: Ongoing cohort study in 794 subjects aged over 70 years from Albacete (Spain). Data were collected on depression risk, frailty, hospitalizations, and covariates. Participants were categorized into six groups. RESULTS: Adjusted hospitalization risk was higher for groups of prefrail/-non depression risk (HR 1.48; 95% confidence interval (CI) 1.16-1.89), prefrail/depression risk (HR 1.73; 95% CI 1.29-2.30), frail/non depression risk (HR 1.79; 95% CI 1.22-2.62), and frail/depression risk (HR 2.12; 95% CI 1.49-3.02), compared with robust/non depression risk group (p<0.01). Frail and prefrail groups presented increased hospitalization density in the first four follow-up years. CONCLUSIONS: Depression risk changes the yearly probabilities of hospitalization in prefrail and frail groups, increasing them in the first years. Depression risk should be monitored in prefrail and frail older adults as an independent risk factor for hospitalization.

Differential Immunoexpression of BRAF/V600E, Senescence Markers, PTEN, and T-type Calcium Channels in Acquired Naevi According to their Histopathological and Dermoscopic Classification.

BRAF/V600E mutation and other cell growth/growth-control mechanisms are involved in naevogenesis and melanomagenesis. Immunoexpression of BRAF/V600E and other molecules (p16, phosphatase and tensin homologue (PTEN), Ki67, hTERT and Cav3.1 and 3.2 calcium channels) were investigated in 80 histopatho-logically and dermoscopically classified acquired naevi. Regarding BRAF/V600E, dysplastic naevi showed lower immunostaining than common naevi, which was significant in comparison with intradermal naevi, which showed the highest BRAF/V600E histoscore. Junctional naevi showed the lowest BRAF/V600E levels. Globular/cobblestone and reticular dermoscopic patterns were consistently associated with high and low BRAF/V600E immunoexpression, respectively, but Zalaudek's peripheral globule pattern (CR/PG) showed the highest BRAF/V600E immunoexpression. Among global patterns, the previously not investigated multicomponent pattern showed the lowest BRAF/V600E immunoexpression. Regarding the remaining biomarkers, new immunohistochemical features were found, in particular p16 and PTEN low expression in multicomponent pattern; and Ki67, hTERT and Cav.3.1 high expression in CR/PG. In conclusion, histopathology and dermoscopy provide complementary information regarding the biology of melanocytic naevi.

Usability, User Experience, and Acceptance Evaluation of CAPACITY: A Technological Ecosystem for Remote Follow-Up of Frailty.

Frailty predisposes older persons to adverse events, and information and communication technologies can play a crucial role to prevent them. CAPACITY provides a means to remotely monitor variables with high predictive power for adverse events, enabling preventative personalized early interventions. This study aims at evaluating the usability, user experience, and acceptance of a novel mobile system to prevent disability. Usability was assessed using the system usability scale (SUS); user experience using the user experience questionnaire (UEQ); and acceptance with the technology acceptance model (TAM) and a customized quantitative questionnaire. Data were collected at baseline (recruitment), and after three and six months of use. Forty-six participants used CAPACITY for six months; nine dropped out, leaving a final sample of 37 subjects. SUS reached a maximum averaged value of 83.68 after six months of use; no statistically significant values have been found to demonstrate that usability improves with use, probably because of a ceiling effect. UEQ, obtained averages scores higher or very close to 2 in all categories. TAM reached a maximum of 51.54 points, showing an improvement trend. Results indicate the success of the participatory methodology, and support user centered design as a key methodology to design technologies for frail older persons. Involving potential end users and giving them voice during the design stage maximizes usability and acceptance.

Malignant deciduoid mesothelioma - to what extent is EUS useful?

An 83-year-old female presented with progressive dyspnea, a non-productive cough and right pleural effusion consistent with exudate. Rapid staining and PCR for mycobacteria, culture and cytology were negative. A thoracic computed tomography (CT) scan revealed a solid lesion, 53 mm in diameter, with an epicenter in the right crus of the diaphragm, both diaphragmatic and hepatic infiltration and compression of the inferior vena cava.

Role of Genetic Variations in the Hepatic Handling of Drugs.

The liver plays a pivotal role in drug handling due to its contribution to the processes of detoxification (phases 0 to 3). In addition, the liver is also an essential organ for the mechanism of action of many families of drugs, such as cholesterol-lowering, antidiabetic, antiviral, anticoagulant, and anticancer agents. Accordingly, the presence of genetic variants affecting a high number of genes expressed in hepatocytes has a critical clinical impact. The present review is not an exhaustive list but a general overview of the most relevant variants of genes involved in detoxification phases. The available information highlights the importance of defining the genomic profile responsible for the hepatic handling of drugs in many ways, such as (i) impaired uptake, (ii) enhanced export, (iii) altered metabolism due to decreased activation of prodrugs or enhanced inactivation of active compounds, and (iv) altered molecular targets located in the liver due to genetic changes or activation/downregulation of alternative/compensatory pathways. In conclusion, the advance in this field of modern pharmacology, which allows one to predict the outcome of the treatments and to develop more effective and selective agents able to overcome the lack of effect associated with the existence of some genetic variants, is required to step forward toward a more personalized medicine.

Candesartan-induced cholestatic hepatitis: a case report.

We present the case of a 55-year-old male that was admitted to our hospital for cholestatic hepatitis. Other causes of hepatitis were excluded with imaging, laboratory and serologic tests. A liver biopsy was performed, which was compatible with toxicity-induced hepatitis. Re-assessing the patient, he mentioned the start of treatment with candesartan cilexetil 3 weeks prior to the onset of the symptoms. Candesartan was withdrawn, with a progressive improvement in cholestasis and complete normalization of liver biochemistry at 6 months. The patient was diagnosed with candesartan-induced cholestatic hepatitis, probable according to CIOMS/RUCAM scale.

New management of hepatolithiasis: Can surgery be avoided? (with video).

BACKGROUND: The presence of hepatolithiasis (HL) is prevalent in eastern countries. It is a clinical entity which is rarely reported in non-surgical series because the standard treatment is the surgical option. Currently, treatment has evolved, with the use of endoscopic techniques being increased and the number of hepatectomies being decreased. SpyGlass™ is a small-calibre endoscopic direct cholangiopancreatoscopy developed to explore and perform procedures in the bile and pancreatic ducts. Single-operator peroral cholangioscopy (POC) is an endoscopic technique useful for treating difficult bile duct stones. AIMS: To assess the usefulness, efficacy, and safety of POC with the SpyGlass™ system in patients with HL. PRIMARY OBJECTIVES: to achieve technical success of the procedure and clinical success of patients with HL. STUDY DESIGN AND PATIENTS: Retrospective, single-centre cohort study of patients with HL from April 2012 to August 2018. SpyGlass™ was chosen in symptomatic patients referred from the surgery unit as the first-line procedure. To perform electrohydraulic lithotripsy (EHL), we used a Northgate Autolith IEHL generator with a 0.66-mm biliary probe. RESULTS: We performed a total of 13 procedures in 7 patients with HL. The mean age was 46 years (range 35-65) and 3/7 of patients were female. We achieved technical success in 5/7 cases (71.4%) and clinical success in 4/7 cases (57%). DISCUSSION: SpyGlass™ is safe and effective in the treatment of HL. With these results, we confirm the need for management of patients with HL in a multidisciplinary team. When the endoscopic approach is the option, this procedure must be performed by experts in advanced endoscopy.

Assessment of specific antibodies as biological indicators of human chronic exposure to microcystins.

Cyanobacteria can produce potent natural toxins known as cyanotoxins. Blooms of cyanobacteria, produced mainly as result of the pollution of water bodies with excessive amounts of phosphorus, represent a severe environmental problem; not only do they affect the normal equilibrium of the aquatic ecosystem but may also affect animal and human health. The occurrence of algal blooms have been increasing globally (it has been recently reported in at least 100 countries) and it has been considered by WHO as an emerging public health issue. The toxic effects of cyanotoxins have been thoroughly demonstrated in laboratory experiments, however, the effects on humans and the extent of these effects have been more difficult to assess. Epidemiological research is difficult as there are no specific symptoms or routine biomarkers to diagnose intoxication with cyanotoxins, in particular those cases associated with chronic exposure. The objectives of this study were to assess the exposure of a population settled near a lake with recurrent cyanobacteria blooms and to investigate the presence of biological markers of chronic exposure to cyanotoxins, in particular the microcystins (MCs). We first investigated the exposure of the population to cyanobacteria by using a questionnaire on how the population used the water and by analyzing water samples for the presence of cyanobacteria and total microcystins (TMCs). Secondly, we investigated the presence of biological indicators by analyzing the biochemical and immunological parameters in sera of the exposed population. The questionnaires and the water analyses revealed that the population under study (n = 47) is exposed to several exposure routes. The biochemical analyses of the sera showed the alteration of at least one hepatic enzyme in 25% of the exposed people, but this cannot be associated solely to MCs exposure. On the contrary, the immunological analyses, which included microcystin-LR specific antibodies IgE and IgG, showed significant differences between the exposed and non-exposed groups. The presence of MCs specific antibodies confirms the exposure to MCs. We propose the study of specific antibodies as a non-complex biomarker to detect chronic exposure to the toxin and to assist epidemiological studies.

Median arcuate ligament syndrome as a cause of intestinal angina.

Median arcuate ligament syndrome (MALS) or celiac artery compression syndrome is caused by extrinsic compression by the median arcuate ligament. It is often asymptomatic, but it can cause intestinal symptoms suggestive of angina. We present a patient with repeated episodes of postprandial pain, nausea and occasional vomiting. After several studies, it was concluded that the symptomatology was attributable to this syndrome. Conservative treatment was performed due to the coexistence of underlying neoplastic disease and low life expectancy.

Chemoresistance and chemosensitization in cholangiocarcinoma.

One of the main difficulties in the management of patients with advanced cholangiocarcinoma (CCA) is their poor response to available chemotherapy. This is the result of powerful mechanisms of chemoresistance (MOC) of quite diverse nature that usually act synergistically. The problem is often worsened by altered MOC gene expression in response to pharmacological treatment. Since CCA includes a heterogeneous group of cancers their genetic signature coding for MOC genes is also diverse; however, several shared traits have been defined. Some of these characteristics are shared with other types of liver cancer, namely hepatocellular carcinoma and hepatoblastoma. An important goal in modern oncologic pharmacology is to develop novel strategies to overcome CCA chemoresistance either by increasing drug specificity, such as in targeted therapies aimed to inhibit receptors with tyrosine kinase activity, or to increase the amounts of active agents inside CCA cells by enhancing drug uptake or reducing efflux through export pumps. This article is part of a Special Issue entitled: Cholangiocytes in Health and Diseaseedited by Jesus Banales, Marco Marzioni, Nicholas LaRusso and Peter Jansen.

Interaction of glucocorticoids with FXR/FGF19/FGF21-mediated ileum-liver crosstalk.

At high doses, glucocorticoids (GC) have been associated with enhanced serum bile acids and liver injury. We have evaluated the effect of GC, in the absence of hepatotoxicity, on FXR/FGF91(Fgf15)/FGF21-mediated ileum-liver crosstalk. Rats and mice (wild type and Fxr-/-, Fgf15-/- and int-Gr-/- strains; the latter with GC receptor (Gr) knockout selective for intestinal epithelial cells), were treated (i.p.) with dexamethasone, prednisolone or budesonide. In both species, high doses of GC caused hepatotoxicity. At a non-hepatotoxic dose, GC induced ileal Fgf15 down-regulation and liver Fgf21 up-regulation, without affecting Fxr expression. Fgf21 mRNA levels correlated with those of several genes involved in glucose and bile acid metabolism. Surprisingly, liver Cyp7a1 was not up-regulated. The expression of factors involved in transcriptional modulation by Fxr and Gr (p300, Drip205, CBP and Smrt) was not affected. Pxr target genes Cyp3a11 and Mrp2 were not up-regulated in liver or intestine. In contrast, the expression of some Pparα target genes in liver (Fgf21, Cyp4a14 and Vanin-1) and intestine (Vanin-1 and Cyp3a11) was altered. In mice with experimental colitis, liver Fgf21 was up-regulated (4.4-fold). HepG2 cells transfection with FGF21 inhibited CYP7A1 promoter (prCYP7A1-Luc2). This was mimicked by pure human FGF21 protein or culture in medium previously conditioned by cells over-expressing FGF21. This response was not abolished by deletion of a putative response element for phosphorylated FGF21 effectors present in prCYP7A1. In conclusion, GC interfere with FXR/FGF19-mediated intestinal control of CYP7A1 expression by the liver and stimulate hepatic secretion of FGF21, which inhibits CYP7A1 promoter through an autocrine mechanism.

Single-operator cholangiopancreatoscopy in pancreatobiliary diseases: clinical experience in a tertiary referral hospital.

BACKGROUND AND AIMS: to assess the usefulness, efficacy and safety of single-operator cholangiopancreatoscopy (SOCP) with the SpyGlass™ system for the management of biliopancreatic diseases. METHODS: a retrospective analysis of patients undergoing SOCP with the SpyGlass™ between September 2008 and April 2016 was performed. Data was obtained from a prospectively-maintained database at a tertiary referral center. The primary study outcomes were technical and complete endoscopic success of the procedure. Two different SpyGlass™ systems were employed; the former is called legacy and the latter, digital system (DS). RESULTS: a total of 107 SOCP procedures in 93 patients performed by a single operator were analyzed. Technical success of the SpyGlass™ examination was achieved in 90/93 (97%) of patients and complete success by resolving the biliopancreatic condition in 82/93 (88%) cases. In indeterminate biliary strictures, a complete success was achieved in 45/52 (85%) of cases. With regard to stone treatment, technical success was achieved in 34/34 (100%) patients and complete success, in 31/34 (91%) cases. Electrohydraulic lithotripsy was applied in 16/34 (47%) of cases. There were a total of 7/93 adverse effects (7.5%). CONCLUSIONS: SOCP is a useful and safe technique for the treatment of biliopancreatic diseases with a low rate of adverse effects. The procedure seems technically demanding and dedication is required.

Efficacy and safety of anticoagulation in non-malignant portal vein thrombosis in patients with liver cirrhosis.

BACKGROUND AND AIM: Treatment for portal vein thrombosis (PVT) is not well established. Nevertheless, anticoagulation therapy can seemingly be used as first-line therapy. However, there are limited data on the role of this treatment in patients with PVT and cirrhosis. We sought to assess the safety and efficacy of anticoagulation therapy in a series of patients with non-malignant PVT and liver cirrhosis. METHODS: We analyzed the data of 32 patients with cirrhosis and PVT between March 2009 and September 2015. All patients received anticoagulation treatment. PVT was diagnosed within the context of biannual hepatocellular carcinoma screening in these patients. RESULTS: Recanalisation was achieved in 23 patients: complete in 17 patients (53.1%) and partial in 6 patients (18.7%). The median time for achieving a complete response was 7 months (95% CI: 6-8). We did not discover any risk factors associated with repermeation (partial or complete). None of the patients presented with thrombosis progression while receiving anticoagulation. Nine patients who achieved complete recanalisation and stopped anticoagulation therapy suffered rethrombosis (52%). There were no differences between the patients who achieved complete or partial recanalisation (35%) and those who did not (33%) in relation to the onset of hepatic events during follow-up. Three patients (9%) presented with bleeding complications: two variceal bleeding episodes and one brain hemorrhage. CONCLUSIONS: In cirrhotic patients with non-malignant PVT, anticoagulation therapy led to partial or complete recanalisation in 70% of patients, with a broad safety profile. Due to the existing rethrombosis rate, long-term anticoagulation should be considered.

Osteopontin regulates the cross-talk between phosphatidylcholine and cholesterol metabolism in mouse liver.

Osteopontin (OPN) is involved in different liver pathologies in which metabolic dysregulation is a hallmark. Here, we investigated whether OPN could alter liver, and more specifically hepatocyte, lipid metabolism and the mechanism involved. In mice, lack of OPN enhanced cholesterol 7α-hydroxylase (CYP7A1) levels and promoted loss of phosphatidylcholine (PC) content in liver; in vivo treatment with recombinant (r)OPN caused opposite effects. rOPN directly decreased CYP7A1 levels through activation of focal adhesion kinase-AKT signaling in hepatocytes. PC content was also decreased in OPN-deficient (OPN-KO) hepatocytes in which de novo FA and PC synthesis was lower, whereas cholesterol (CHOL) synthesis was higher, than in WT hepatocytes. In vivo inhibition of cholesterogenesis normalized liver PC content in OPN-KO mice, demonstrating that OPN regulates the cross-talk between liver CHOL and PC metabolism. Matched liver and serum samples showed a positive correlation between serum OPN levels and liver PC and CHOL concentration in nonobese patients with nonalcoholic fatty liver. In conclusion, OPN regulates CYP7A1 levels and the metabolic fate of liver acetyl-CoA as a result of CHOL and PC metabolism interplay. The results suggest that CYP7A1 is a main axis and that serum OPN could disrupt liver PC and CHOL metabolism, contributing to nonalcoholic fatty liver disease progression in nonobese patients.