Therapeutic targeting of adipose tissue macrophages ameliorates liver fibrosis in non-alcoholic fatty liver disease.

Background & Aims: : The accumulation of adipose tissue macrophages (ATMs) in obesity has been associated with hepatic injury. However, the contribution of ATMs to hepatic fibrosis in non-alcoholic fatty liver disease (NAFLD) remains to be elucidated. Herein, we investigate the relationship between ATMs and liver fibrosis in patients with patients with NAFLD and evaluate the impact of modulation of ATMs over hepatic fibrosis in an experimental non-alcoholic steatohepatitis (NASH) model. Methods: Adipose tissue and liver biopsies from 42 patients with NAFLD with different fibrosis stages were collected. ATMs were characterised by immunohistochemistry and flow cytometry and the correlation between ATMs and liver fibrosis stages was assessed. Selective modulation of the ATM phenotype was achieved by i.p. administration of dextran coupled with dexamethasone in diet-induced obesity and NASH murine models. Chronic administration effects were evaluated by histology and gene expression analysis in adipose tissue and liver samples. In vitro crosstalk between human ATMs and hepatic stellate cells (HSCs) and liver spheroids was performed. Results: Patients with NAFLD presented an increased accumulation of pro-inflammatory ATMs that correlated with hepatic fibrosis. Long-term modulation of ATMs significantly reduced pro-inflammatory phenotype and ameliorated adipose tissue inflammation. Moreover, ATMs modulation was associated with an improvement in steatosis and hepatic inflammation and significantly reduced fibrosis progression in an experimental NASH model. In vitro, the reduction of the pro-inflammatory phenotype of human ATMs with dextran-dexamethasone treatment reduced the secretion of inflammatory chemokines and directly attenuated the pro-fibrogenic response in HSCs and liver spheroids. Conclusions: Pro-inflammatory ATMs increase in parallel with fibrosis degree in patients with NAFLD and their modulation in an experimental NASH model improves liver fibrosis, uncovering the potential of ATMs as a therapeutic target to mitigate liver fibrosis in NAFLD. Impact and implications: We report that human adipose tissue pro-inflammatory macrophages correlate with hepatic fibrosis in non-alcoholic fatty liver disease (NAFLD). Furthermore, the modulation of adipose tissue macrophages (ATMs) by dextran-nanocarrier conjugated with dexamethasone shifts the pro-inflammatory phenotype of ATMs to an anti-inflammatory phenotype in an experimental murine model of non-alcoholic steatohepatitis. This shift ameliorates adipose tissue inflammation, hepatic inflammation, and fibrosis. Our results highlight the relevance of adipose tissue in NAFLD pathophysiology and unveil ATMs as a potential target for NAFLD.

Ductular reaction-associated neutrophils promote biliary epithelium proliferation in chronic liver disease.

BACKGROUND & AIMS: Ductular reaction expansion is associated with poor prognosis in patients with advanced liver disease. However, the mechanisms promoting biliary cell proliferation are largely unknown. Here, we identify neutrophils as drivers of biliary cell proliferation and the defective wound-healing response. METHODS: The intrahepatic localization of neutrophils was evaluated in patients with chronic liver disease. Neutrophil dynamics were analyzed by intravital microscopy and neutrophil-labeling assays in DDC-treated mice. Neutrophil depletion or inhibition of recruitment was achieved using a Ly6g antibody or a CXCR1/2 inhibitor, respectively. Mice deficient in PAD4 (peptidyl arginine deiminase 4) and ELANE/NE (neutrophil elastase) were used to investigate the mechanisms underlying ductular reaction expansion. RESULTS: In this study we describe a population of ductular reaction-associated neutrophils (DRANs), which are in direct contact with biliary epithelial cells in chronic liver diseases and whose numbers increased in parallel with disease progression. We show that DRANs are immobilized at the site of ductular reaction for a prolonged period of time. In addition, liver neutrophils display a unique phenotypic and transcriptomic profile, showing a decreased phagocytic capacity and increased oxidative burst. Depletion of neutrophils or inhibition of their recruitment reduces DRANs and the expansion of ductular reaction, while mitigating liver fibrosis and angiogenesis. Mechanistically, neutrophils deficient in PAD4 and ELANE abrogate neutrophil-induced biliary cell proliferation, thus indicating the role of neutrophil extracellular traps and elastase release in ductular reaction expansion. CONCLUSIONS: Overall, our study reveals the accumulation of DRANs as a hallmark of advanced liver disease and a potential therapeutic target to mitigate ductular reaction and the maladaptive wound-healing response. IMPACT AND IMPLICATIONS: Our results indicate that neutrophils are highly plastic and can have an extended lifespan. Moreover, we identify a new role of neutrophils as triggers of expansion of the biliary epithelium. Overall, the results of this study indicate that ductular reaction-associated neutrophils (or DRANs) are new players in the maladaptive tissue-healing response in chronic liver injury and may be a potential target for therapeutic interventions to reduce ductular reaction expansion and promote tissue repair in advanced liver disease.

Hepatocyte Dedifferentiation Profiling In Alcohol-Related Liver Disease Identifies CXCR4 As A Driver Of Cell Reprogramming.

Background and Aims: Loss of hepatocyte identity is associated with impaired liver function in alcohol-related hepatitis (AH). In this context, hepatocyte dedifferentiation gives rise to cells with a hepatobiliary (HB) phenotype expressing biliary and hepatocytes markers and showing immature features. However, the mechanisms and the impact of hepatocyte dedifferentiation in liver disease are poorly understood. Methods: HB cells and ductular reaction (DR) cells were quantified and microdissected from liver biopsies from patients with alcohol-related liver disease (ALD). Hepatocyte- specific overexpression or deletion of CXCR4, and CXCR4 pharmacological inhibition were assessed in mouse liver injury. Patient-derived and mouse organoids were generated to assess plasticity. Results: Here we show that HB and DR cells are increased in patients with decompensated cirrhosis and AH, but only HB cells correlate with poor liver function and patients' outcome. Transcriptomic profiling of HB cells revealed the expression of biliary-specific genes and a mild reduction of hepatocyte metabolism. Functional analysis identified pathways involved in hepatocyte reprogramming, inflammation, stemness and cancer gene programs. CXCR4 pathway was highly enriched in HB cells, and correlated with disease severity and hepatocyte dedifferentiation. In vitro , CXCR4 was associated with biliary phenotype and loss of hepatocyte features. Liver overexpression of CXCR4 in chronic liver injury decreased hepatocyte specific gene expression profile and promoted liver injury. CXCR4 deletion or its pharmacological inhibition ameliorated hepatocyte dedifferentiation and reduced DR and fibrosis progression. Conclusions: This study shows the association of hepatocyte dedifferentiation with disease progression and poor outcome in AH. Moreover, the transcriptomic profiling of HB cells revealed CXCR4 as a new driver of hepatocyte-to-biliary reprogramming and as a potential therapeutic target to halt hepatocyte dedifferentiation in AH. Lay summary: Here we describe that hepatocyte dedifferentiation is associated with disease severity and a reduced synthetic capacity of the liver. Moreover, we identify the CXCR4 pathway as a driver of hepatocyte dedifferentiation and as a therapeutic target in alcohol-related hepatitis.

Hepatocyte dedifferentiation profiling in alcohol-related liver disease identifies CXCR4 as a driver of cell reprogramming.

BACKGROUND & AIMS: Loss of hepatocyte identity is associated with impaired liver function in alcohol-related hepatitis (AH). In this context, hepatocyte dedifferentiation gives rise to cells with a hepatobiliary (HB) phenotype expressing biliary and hepatocyte markers and showing immature features. However, the mechanisms and impact of hepatocyte dedifferentiation in liver disease are poorly understood. METHODS: HB cells and ductular reaction (DR) cells were quantified and microdissected from liver biopsies from patients with alcohol-related liver disease (ArLD). Hepatocyte-specific overexpression or deletion of C-X-C motif chemokine receptor 4 (CXCR4), and CXCR4 pharmacological inhibition were assessed in mouse liver injury. Patient-derived and mouse organoids were generated to assess plasticity. RESULTS: Here, we show that HB and DR cells are increased in patients with decompensated cirrhosis and AH, but only HB cells correlate with poor liver function and patients' outcome. Transcriptomic profiling of HB cells revealed the expression of biliary-specific genes and a mild reduction of hepatocyte metabolism. Functional analysis identified pathways involved in hepatocyte reprogramming, inflammation, stemness, and cancer gene programs. The CXCR4 pathway was highly enriched in HB cells and correlated with disease severity and hepatocyte dedifferentiation. In vitro, CXCR4 was associated with a biliary phenotype and loss of hepatocyte features. Liver overexpression of CXCR4 in chronic liver injury decreased the hepatocyte-specific gene expression profile and promoted liver injury. CXCR4 deletion or its pharmacological inhibition ameliorated hepatocyte dedifferentiation and reduced DR and fibrosis progression. CONCLUSIONS: This study shows the association of hepatocyte dedifferentiation with disease progression and poor outcome in AH. Moreover, the transcriptomic profiling of HB cells revealed CXCR4 as a new driver of hepatocyte-to-biliary reprogramming and as a potential therapeutic target to halt hepatocyte dedifferentiation in AH. IMPACT AND IMPLICATIONS: Here, we show that hepatocyte dedifferentiation is associated with disease severity and a reduced synthetic capacity of the liver. Moreover, we identify the CXCR4 pathway as a driver of hepatocyte dedifferentiation and as a therapeutic target in alcohol-related hepatitis. Therefore, this study reveals the importance of preserving strict control over hepatocyte plasticity in order to preserve liver function and promote tissue repair.

What drives the decision to optimise biological treatment in children and youngsters with juvenile idiopathic arthritis? A discrete-choice experiment.

OBJECTIVE: To analyse factors involved in the decision to optimise biologics in juvenile idiopathic arthritis. METHODS: A "discrete-choice" methodology was used. In a nominal group meeting, factors which may influence physicians' decisions to optimise biological dose were identified, together with decision nodes. 1000Minds® was used to create multiple fictitious clinical scenarios based on the factors identified, and to deploy surveys that were sent to a panel of experts. These experts decided for each item which of two clinical scenarios prompted them to optimise the dose of biologic. A conjoint analysis was carried out, and the partial-value functions and the weights of relative importance calculated. RESULTS: In the nominal group, three decision nodes were identified: (1) time to decide; (2) to maintain/reduce or prolong interval; (3) what drug to reduce. The factors elicited were different for each node and included patient and drug attributes. The presence of macrophage activation syndrome (MAS), systemic involvement, or subclinical inflammation made the decision easier (highest weights). The presence of joints of difficult control and year of debut influenced the decision in some but not all, and in different directions. Immunogenicity, adherence, and concomitant treatments were also aspects taken into account. CONCLUSIONS: The decision to optimise the dose of biological therapy in children and youngster can be divided into several nodes, and the factors, both patient and therapy-related, leading to the decision can be detailed. These decisions taken by experts may be transported to practice, study designs, and guidelines.

Effects of the use of autobiographical photographs on emotional induction in older adults: a systematic review.

There is evidence to support the positive contribution of autobiographical recall based techniques on individuals' quality of life, mood and cognitive functioning. In this review, we analyzed the effects of the use of personal photographs in interventions based on autobiographical memory in older people with and without cognitive impairment. The PRISMA guidelines for systematic reviews were followed. The search was carried out in the electronic databases Web of Sciences (WOS), Medline (PubMed), SCIELO and PsycInfo (American Psychological Association). The articles based on clinical trials selected were evaluated using the PEDRo scale, which is specific to this type of article. Of the 1098 articles initially found, 6 met the inclusion criteria. The final articles focused their intervention on the use of autobiographical photographs as a means of stimulation. The results show that the use of photographs in different autobiographical recall stimulation techniques is associated with higher scores on well-being and quality of life, as well as with improvements in personal identity and cognitive functioning. This suggests that using personal photographs shows promise in enhancing the effect of these types of interventions in healthy or cognitively impaired older adults.

[Seroprevalence of Hepatitis B virus (HBV), Hepatitis C virus (HCV) and Human Immunodeficiency Virus (HIV) in a sample of patients diagnosed with Alcohol User Disorder in Albacete (Spain).] / Seroprevalencia del virus de la hepatitis B, del virus de la hepatitis C y del virus de la inmunodeficiencia humana en una muestra de pacientes diagnosticados de trastorno por consumo de alcohol en Albacete.

OBJECTIVE: Studies report a higher prevalence of HCV, HBV and HIV in the population with Alcohol Use Disorder. Our objective is to determine the seroprevalence for hepatitis B and C and the human immunodeficiency virus (HIV) in patients with alcohol use disorder, as well as to determine if there is a correct serological screening of these. METHODS: Retrospective study of 204 patients with Alcohol Use Disorder followed up in the Addictive Behaviors Unit from Albacete. They started treatment from 2013-2014 onwards, the last patient was recruited in December of 2017. RESULTS: Our sample has 160 men (78.4%) and 44 women (21.6%). The median age is 46.2 years. 161 patients (78.9%) had HIV serology and it was negative in all. 146 patients (71.6%) had negative hepatitis B serology and 12 (5.9%) were vaccinated. 36 patients (17.6%) had no hepatitis B serology performed. In 10 patients, hepatitis B serology was positive (4.9%; 95% CI 1.9-7.9%). 159 patients (77.9%) had negative hepatitis C serology, and 37 patients had not performed it (18.1%). 8 patients had positive serology (prevalence of 3.9%; 95% CI 1.2-6.6%). Excluding patients who did not have serology, seroprevalence was 5.9% (95% CI 2.3-9.6%) and 4.8% (95% CI 1.5-8.1%) respectively. Failure to perform serologies was associated with older age and less weekly Standard Drink Units (SDUs) consumption. CONCLUSIONS: The seroprevalence of HBV and HCV in patients with alcohol use disorder is high. It should be ensured that 100% of patients with alcohol use disorder undergo HCV, HIV, HBV serology, reducing the loss of diagnostic opportunities.

OBJETIVO: Estudios relatan mayor prevalencia de VHC, VHB y VIH en población con Trastorno de Consumo de Alcohol. El objetivo de nuestro estudio fue determinar la seroprevalencia para Virus de Hepatitis B (VHB), Virus de Hepatitis C (VHC) y el Virus de la Inmunodeficiencia Humana (VIH) en pacientes con trastorno por consumo de alcohol, así como determinar si existe un correcto cribaje serológico de éstos. METODOS: Estudio retrospectivo de 204 pacientes con trastorno por consumo de alcohol seguidos en la unidad de conductas adictivas de Albacete, que iniciaron tratamiento desde el año 2013-2014 en adelante, el último paciente fue reclutado en diciembre de 2017. RESULTADOS: Nuestra muestra tiene 160 hombres (78,4%) y 44 mujeres (21,6%). La mediana de edad es 46,2 años. 161 pacientes (78,9%) tenían serología de VIH y fue negativa en todos. 146 pacientes (71,6%) tenían serología negativa de hepatitis B y 12 (5,9%) estaban vacunados. 36 pacientes (17,6%) no tenían serología realizada de hepatitis B. En 10 pacientes la serología de hepatitis B fue positiva (4,9%; IC95% 1,9-7,9%). 159 pacientes (77,9%) tenían serología negativa de hepatitis C, y 37 pacientes no la tenían realizada (18,1%). 8 pacientes tenían serología positiva (prevalencia de 3,9%; IC95% 1,2-6,6%). Excluyendo a los pacientes que no tenían serología la seroprevalencia fue de 5,9% (IC95% 2,3-9,6%) y de 4.8% (IC95% de 1,5-8,1%) respectivamente. La no realización de serologías se asoció a mayor edad y un menor consumo de Unidades de Bebida Estándar (UBEs) por semana. CONCLUSIONES: La seroprevalencia de VHB y VHC en pacientes con trastorno por consumo de alcohol es alta. Debería asegurarse que el 100% de pacientes con trastorno de consumo de alcohol, se realicen serologías de VHC, VIH, VHB disminuyendo pérdida de oportunidades diagnósticas.

The link between non-suicidal self-injury (NSSI), body image and activity on social networking sites among female adolescents with an eating disorder.

The link between engagement in non-suicidal self-injury (NSSI)-related activities on SNS (e.g. viewing, commenting, sharing and uploading NSSI content) with body image and NSSI behaviour remains under researched in adolescents with EDs. The main aim of the current research was to examine associations between NSSI-related activities on SNS with body image and NSSI behaviour severity among female adolescents diagnosed with an ED. A total of 52 female adolescents (mean age = 15.35 years, SD = 1.49) diagnosed with an ED completed self-report questionnaires related to NSSI behaviour, SNS usage and body image. Participants were divided into two groups: low NSSI behaviour severity (from 0 to 10 NSSI behaviours; n = 28) and high NSSI behaviour severity (more than ten NSSI behaviours; n = 24). Within the high NSSI severity group, individuals that comment and share NSSI online content significantly reported higher negative body image. A hierarchical binary logistic regression showed that the frequency of NSSI online content on SNS emerged as significant predictor of NSSI behaviour severity within last year after controlling for body image and searching for ED content on SNS. Our findings suggest that not only searching for ED content, but also being daily engaged in NSSI online activities may increase the risk of NSSI behaviour severity in female adolescents with EDs.

Common Laboratory Parameters Are Useful for Screening for Alcohol Use Disorder: Designing a Predictive Model Using Machine Learning.

The diagnosis of alcohol use disorder (AUD) remains a difficult challenge, and some patients may not be adequately diagnosed. This study aims to identify an optimum combination of laboratory markers to detect alcohol consumption, using data science. An analytical observational study was conducted with 337 subjects (253 men and 83 women, with a mean age of 44 years (10.61 Standard Deviation (SD)). The first group included 204 participants being treated in the Addictive Behaviors Unit (ABU) from Albacete (Spain). They met the diagnostic criteria for AUD specified in the Diagnostic and Statistical Manual of mental disorders fifth edition (DSM-5). The second group included 133 blood donors (people with no risk of AUD), recruited by cross-section. All participants were also divided in two groups according to the WHO classification for risk of alcohol consumption in Spain, that is, males drinking more than 28 standard drink units (SDUs) or women drinking more than 17 SDUs. Medical history and laboratory markers were selected from our hospital's database. A correlation between alterations in laboratory markers and the amount of alcohol consumed was established. We then created three predicted models (with logistic regression, classification tree, and Bayesian network) to detect risk of alcohol consumption by using laboratory markers as predictive features. For the execution of the selection of variables and the creation and validation of predictive models, two tools were used: the scikit-learn library for Python, and the Weka application. The logistic regression model provided a maximum AUD prediction accuracy of 85.07%. Secondly, the classification tree provided a lower accuracy of 79.4%, but easier interpretation. Finally, the Naive Bayes network had an accuracy of 87.46%. The combination of several common biochemical markers and the use of data science can enhance detection of AUD, helping to prevent future medical complications derived from AUD.

Hedgehog Signalling Modulates Immune Response and Protects against Experimental Autoimmune Encephalomyelitis.

The Hedgehog (Hh) pathway is essential for the embryonic development and homeostatic maintenance of many adult tissues and organs. It has also been associated with some functions of the innate and adaptive immune system. However, its involvement in the immune response has not been well determined. Here we study the role of Hh signalling in the modulation of the immune response by using the Ptch-1-LacZ+/- mouse model (hereinafter referred to as ptch+/-), in which the hemizygous inactivation of Patched-1, the Hh receptor gene, causes the constitutive activation of Hh response genes. The in vitro TCR stimulation of spleen and lymph node (LN) T cells showed increased levels of Th2 cytokines (IL-4 and IL-10) in ptch+/-cells compared to control cells from wild-type (wt) littermates, suggesting that the Th2 phenotype is favoured by Hh pathway activation. In addition, CD4+ cells secreted less IL-17, and the establishment of the Th1 phenotype was impaired in ptch+/- mice. Consistently, in response to an inflammatory challenge by the induction of experimental autoimmune encephalomyelitis (EAE), ptch+/- mice showed milder clinical scores and more minor spinal cord damage than wt mice. These results demonstrate a role for the Hh/ptch pathway in immune response modulation and highlight the usefulness of the ptch+/- mouse model for the study of T-cell-mediated diseases and for the search for new therapeutic strategies in inflammatory diseases.

Ductular reaction promotes intrahepatic angiogenesis through Slit2-Roundabout 1 signaling.

BACKGROUND AND AIMS: Ductular reaction (DR) expands in chronic liver diseases and correlates with disease severity. Besides its potential role in liver regeneration, DR plays a role in the wound-healing response of the liver, promoting periductular fibrosis and inflammatory cell recruitment. However, there is no information regarding its role in intrahepatic angiogenesis. In the current study we investigated the potential contribution of DR cells to hepatic vascular remodeling during chronic liver disease. APPROACH AND RESULTS: In mouse models of liver injury, DR cells express genes involved in angiogenesis. Among angiogenesis-related genes, the expression of Slit2 and its receptor Roundabout 1 (Robo1) was localized in DR cells and neoangiogenic vessels, respectively. The angiogenic role of the Slit2-Robo1 pathway in chronic liver disease was confirmed in ROBO1/2-/+ mice treated with 3,5-diethoxycarbonyl-1,4-dihydrocollidine, which displayed reduced intrahepatic neovascular density compared to wild-type mice. However, ROBO1/2 deficiency did not affect angiogenesis in partial hepatectomy. In patients with advanced alcohol-associated disease, angiogenesis was associated with DR, and up-regulation of SLIT2-ROBO1 correlated with DR and disease severity. In vitro, human liver-derived organoids produced SLIT2 and induced tube formation of endothelial cells. CONCLUSIONS: Overall, our data indicate that DR expansion promotes angiogenesis through the Slit2-Robo1 pathway and recognize DR cells as key players in the liver wound-healing response.

Tocilizumab in refractory Caucasian Takayasu's arteritis: a multicenter study of 54 patients and literature review.

OBJECTIVE: To assess the efficacy and safety of tocilizumab (TCZ) in Caucasian patients with refractory Takayasu's arteritis (TAK) in clinical practice. METHODS: A multicenter study of Caucasian patients with refractory TAK who received TCZ. The outcome variables were remission, glucocorticoid-sparing effect, improvement in imaging techniques, and adverse events. A comparative study between patients who received TCZ as monotherapy (TCZMONO) and combined with conventional disease modifying anti-rheumatic drugs (cDMARDs) (TCZCOMBO) was performed. RESULTS: The study comprised 54 patients (46 women/8 men) with a median [interquartile range (IQR)] age of 42.0 (32.5-50.5) years. TCZ was started after a median (IQR) of 12.0 (3.0-31.5) months since TAK diagnosis. Remission was achieved in 12/54 (22.2%), 19/49 (38.8%), 23/44 (52.3%), and 27/36 (75%) patients at 1, 3, 6, and 12 months, respectively. The prednisone dose was reduced from 30.0 mg/day (12.5-50.0) to 5.0 (0.0-5.6) mg/day at 12 months. An improvement in imaging findings was reported in 28 (73.7%) patients after a median (IQR) of 9.0 (6.0-14.0) months. Twenty-three (42.6%) patients were on TCZMONO and 31 (57.4%) on TCZCOMBO: MTX (n = 28), cyclosporine A (n = 2), azathioprine (n = 1). Patients on TCZCOMBO were younger [38.0 (27.0-46.0) versus 45.0 (38.0-57.0)] years; difference (diff) [95% confidence interval (CI) = -7.0 (-17.9, -0.56] with a trend to longer TAK duration [21.0 (6.0-38.0) versus 6.0 (1.0-23.0)] months; diff 95% CI = 15 (-8.9, 35.5), and higher c-reactive protein [2.4 (0.7-5.6) versus 1.3 (0.3-3.3)] mg/dl; diff 95% CI = 1.1 (-0.26, 2.99). Despite these differences, similar outcomes were observed in both groups (log rank p = 0.862). Relevant adverse events were reported in six (11.1%) patients, but only three developed severe events that required TCZ withdrawal. CONCLUSION: TCZ in monotherapy, or combined with cDMARDs, is effective and safe in patients with refractory TAK of Caucasian origin.

Directed differentiation of human induced pluripotent stem cells to hepatic stellate cells.

Hepatic stellate cells (HSCs) are nonparenchymal liver cells responsible for extracellular matrix homeostasis and are the main cells involved in the development of liver fibrosis following injury. The lack of reliable sources of HSCs has hence limited the development of complex in vitro systems to model liver diseases and toxicity. Here we describe a protocol to differentiate human induced pluripotent stem cells (iPSCs) into hepatic stellate cells (iPSC-HSCs). The protocol is based on the addition of several growth factors important for liver development sequentially over 12 d. iPSC-HSCs present phenotypic and functional characteristics of primary HSCs and can be expanded or frozen and used to perform high-throughput in vitro studies. We also describe how to coculture iPSC-HSCs with hepatocytes, which self-assemble into three-dimensional (3D) hepatic spheroids. This protocol enables the generation of HSC-like cells for in vitro modeling and drug screening studies.

Osteoarthritis-related biomarkers profile in chronic anterior cruciate ligament injured knee.

OBJECTIVE: Increasing evidence has shown the involvement of the innate immune system and inflammatory response in osteoarthritis (OA) pathogenesis; however, anterior cruciate ligament (ACL) tears are recognized risk factors for development of post-traumatic OA. We investigated (1) whether inflammatory mediators involved in OA pathogenesis are also present at significant concentrations in the knee joint sometime after ACL complete tear and may be considered as prognostic biomarkers of progression to secondary OA; and (2) whether quantification in serum may surrogate synovial fluid (SF) measurements in both cases. METHODS: Thirty-seven end-stage OA patients and 33 patients with ACL complete tear that were included on the waiting list for knee surgery were consecutively recruited. Serum and SF samples were taken before surgery, and tumor necrosis factor-alpha, (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), matrix metalloproteinase-1 (MMP1), matrix metalloproteinase-3 (MMP3), tissue inhibitor of metalloproteinase-1 (TIMP1), bone morphogenetic protein-7 (BMP7), regulated upon activation normal t-cell expressed and secreted (RANTES), cytokine interferon-γ-induced protein 10 (IP-10) and heat shock protein family A (Hsp70) member 1A (HSPA1A) were quantified by enzyme-linked immunosorbent assay (ELISA.) Normally distributed data were compared using a one-way analysis of variance (ANOVA) test. Data not normally distributed were analyzed using a nonparametric Mann-Whitney rank sum test. Differences in means were compared using a Student's t-test. Correlations were analyzed using Pearson's coefficient of variation. RESULTS: Eighty-seven percent of patients with OA and 86% of those with ACL tear had quantifiable levels of biomarkers in SF. SF levels of IL-6, IL-8, MMP1, MMP3, RANTES, IP-10, BMP7 and HSPA1A were significantly lower in ACL injury knees compared with those with OA, but much higher than those reported in control subjects. Serum levels of IL-6, IP-10, and MMP1 were also lower in patients with ACL tears, who had, however, significantly higher TNF-α, HSPA1A, and TIMP1 levels when compared with OA patients. Levels of biomarkers tested in serum and SF samples were significantly different. CONCLUSIONS: Our data propose that cytokines IL-6 and IL-8 and the chemokines RANTES, IP-10, MMP1, MMP3, and HSPA1A may be involved in the inflammatory process leading to synovitis, the central lesion in OA onset and development; persistent high levels of these substances sometime after ACL injury suggest that they could play an etiopathogenic role in the maintenance of the inflammatory environment leading to post-traumatic OA. Serum biomarker levels do not appear to faithfully reflect what occurs inside the joint. Thus, most biomarkers cannot yet be considered as useful inflammatory biomarkers of knee joint diseases.

Functioning of autobiographical memory specificity and self-defining memories in people with cancer diagnosis.

OBJECTIVES: Cognitive and emotional disturbances have been associated with the diagnosis and treatment of cancer. Autobiographical memory is one of the specific cognitive processes affected during this disease. The current study had two main aims: (1) to compare the functioning of autobiographical memory specificity and its related variables (executive functioning, depression and perceived stress) in a group of persons with cancer and a control group; and (2) to analyze whether the experience of cancer evolved into a self-defining memory in the sample of participants diagnosed with this disease. METHOD: The study sample comprised 62 participants, 31 in the group with a cancer diagnosis and 31 in the control group. Autobiographical memory specificity, executive functions, depression, stress and self-defining memory were evaluated in the current study. RESULTS: Depressive symptomatology and reduced executive functioning, but not perceived stress levels, are related and are predictors of autobiographical memory specificity. In addition, the identified characteristics of the self-defining memories were associated with the cancer experience as a threat to physical integrity and an awareness of the meaning of life. CONCLUSION: This emerging research line is especially important in view of its possible impacts on patients' well-being, due to the importance of psychological processes in cancer disease.

How effective are films in inducing positive and negative emotional states? A meta-analysis.

Meta-analyses and reviews on emotion research have shown the use of film clips to be one of the most effective methods of mood induction. Nonetheless, the effectiveness of this method when positive, negative and neutral emotional targets are studied under similar experimental conditions is currently unknown. This comprehensive meta-analysis included only studies that implemented neutral, positive and negative mood inductions to evaluate the effectiveness of the film clip method as a mood induction procedure. In addition, several factors related to the films, sample and experimental procedure used, the number of emotional categories, for example, or the number of film clips watched, were included to study their influence on the effectiveness of this mood induction procedure. Forty-five studies were included with 6675 participants and 12 possible moderator variables according to the sample and the research procedure. Our findings suggest that film clips are especially powerful in inducing negative mood states (Hedges' g for valence = -1.49 and for arousal = -1.77) although they are also effective inducers of positive mood states (Hedges' g for valence of = . -1.22 and for arousal = -1.34). Additionally, this meta-analysis reveals that variables, such as the number of emotional categories or the type of stimulus used to measure the baseline, should be considered.

Handling Parathormone Receptor Type 1 in Skeletal Diseases: Realities and Expectations of Abaloparatide.

Musculoskeletal disorders represent an elevated socioeconomic burden for developed aging societies. Osteoporosis (OP) has been treated with antiresorptive therapies or with teriparatide that was until recently the only anabolic therapy. However, approval of osteoporosis treatment in postmenopausal women with abaloparatide, which is an analog of parathyroid hormone-related peptide (PTHrP), has created a new alternative for OP management. The success of this new treatment is related to differential mechanisms of activation of PTH receptor type 1 (PTH1R) by abaloparatide and PTH. Here, we address the distinguishing mechanisms of PTH1R activation; the effects of PTH1R stimulation in osteoblast, osteocytes, and chondrocytes; the differences between PTH and abaloparatide actions on PTH1R; potential safety concerns; and future perspectives about abaloparatide use in other musculoskeletal disorders.

Ability emotional intelligence in young people and older adults with and without depressive symptoms, considering gender and educational level.

BACKGROUND: There is little research on differences in Emotional Intelligence (EI) ability at different stages of adult development. The few published studies tend not to use older adult samples. Previous studies on EI ability and age have shown contradictory results. Our main objective was to evaluate results in EI ability across different stages of adult development, taking into account gender, depressive symptoms, and educational level. METHODS: We interviewed 166 participants (108 women), 66 of whom were aged 18-30 years, 53 aged 31-60 years, and 40 aged 61-76 years. All were either working or enrolled in colleges at the time of the study. The assessment tools used were the Mayer-Salovey-Caruso Emotional Intelligence Test (MSCEIT), a test that assesses performance-based EI, and the Centre for Epidemiologic Studies-Depression (CES-D) scale, a tool to evaluate depressive symptoms. RESULTS: Young people, women, and participants with a higher educational level achieved higher scores on the MSCEIT. Additionally, depressive symptomatology was only partially associated with the MSCEIT (i.e., with the using emotions branch). However, a subsequent joint analysis of the independent effects of variables age, gender, educational level, and depressive symptomatology and their interactions on MSCEIT total suggests that only educational level and depressive symptomatology were associated with EI ability, with the direct relationship between age and gender with MSCEIT disappearing. Additionally, our study indicated an interaction effect between age and depressive symptoms, showing that participants in age cohorts 18-30 and 31-60 and without depressive symptoms have a higher EI ability. DISCUSSION: Our study suggests that the direct effects of age and gender on EI ability across adult development, using a wide age range, can change or disappear when effects of educational level and depressive symptomatology, and their interactions, are controlled for. Our results also suggest that EI ability is a protective factor against depression in some age cohorts. This novel aspect of our study does not appear in the previous literature. However, prospective studies are needed to verify these findings and examine whether other psychological variables could determine the relations between age, gender and EI ability across adult development.

Gene expression, protein profiling, and chemotactic activity of infrapatellar fat pad mesenchymal stem cells in pathologies of the knee joint.

The infrapatellar fat pad (IPFP) is a periarticular adipose knee tissue. This tissue contains a large number of mesenchymal stem cells (MSCs). In the present work, we wanted to study the IPFP MSCs and their relationship and differences in two groups, anterior cruciate ligament (ACL) ruptures knees and ostheoarthrosis (OA). The IPFP of 42 patients with OA or ACL rupture were analyzed. Isolation, primary culture, and a genetic and proteomic study of MSCs from IPFP were performed. Gene expression of IL-6, tumor necrosis factor (TNF), IL-8, HSPA1A (Hsp70), CXCL10, RANTES, MMP1, MMP3, TIMP1, and BMP7 was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). We analyzed MSCs from from 12 diferents patients in two cellular pools (6 from AO disease and 6 from ALC rupture to form two cell pool), for the iTRAQ Proteomic Assay. The conditional media were used in quantitative analysis of MSC soluble factors by Luminex and for de migration assay. A higher gene expression of IL-6, TNF, CXCL10, RANTES, and MMP1 and OPG in MSCs from OA versus ACL (p < 0.05) was observed. Conversely HSPA1A, TIMP1, and RANKL showed a significant lower expression in OA-MSCs (p < 0.05). In the secretome analysis, adipsin and visfantin levels in the supernatants from OA-MSCs were lower (p < 0.05) respect to ACL-MSCs. Also, the monocytic cells migrated two-folds in the presence of conditioned media from OA-MSCs patients versus patients with ACL-MSC. The infrapatellar pad should be considered as an adipose tissue capable of producing and excreting inflammatory mediators directly in the knee joint, influencing the development and progression of knee joint pathologies.

Laser capture microdissection: techniques and applications in liver diseases.

Routine transcriptomic and proteomic analysis are usually performed at a whole organ or tissue level. These approaches provide an average readout of all cell types present within the tissue but do not allow differentiating the profile of specific cell populations. Laser capture microdissection (LCM) constitutes an excellent tool to isolate cell populations or areas of interest within a tissue. By direct visualization, the selected area is excised by a laser and can be further processed for a variety of downstream analyses. This technology has been widely used in the study of liver diseases, from DNA and RNA sequencing to mass spectrometry. However, LCM also has important limitations. To ensure the best integrity of the molecule of interest, optimal tissue preservation, careful tissue sectioning, and optimization of the staining procedure are required. The present review provides a description of the LCM technology, including tips and technical recommendations to perform the procedure, as well as an overview of studies using LCM technology in the field of liver disease.