Ammonium quantification in human plasma by proton nuclear magnetic resonance for staging of liver fibrosis in alcohol-related liver disease and nonalcoholic fatty liver disease.

Liver fibrosis staging is a key element driving the prognosis of patients with chronic liver disease. Currently, biopsy is the only technique capable of diagnosing liver fibrosis in patients with alcohol-related liver disease (ArLD) and nonalcoholic fatty liver disease (NAFLD) unequivocally. Noninvasive (e.g. plasma-based) biomarker assays are attractive tools to diagnose and stage disease, yet must prove that they are reliable and sensitive to be used clinically. Here, we demonstrate proton nuclear magnetic resonance as a method to rapidly quantify the endogenous concentration of ammonium ions from human plasma extracts and show their ability to report upon early and advanced stages of ArLD and NAFLD. We show that, irrespective of the disease etiology, ammonium concentration is a more robust and informative marker of fibrosis stage than current clinically assessed blood hepatic biomarkers. Subject to validation in larger cohorts, the study indicates that the method can provide accurate and rapid staging of ArLD and NAFLD without the need for an invasive biopsy.

Sequential changes in urinary biomarker levels in patients with cirrhosis and severe hepatorenal syndrome.

Whether tubular injury develops in patients with acute kidney injury owing to hepatorenal syndrome (AKI-HRS) is controversial. We performed repeated measurements of biomarkers of tubular injury during a 14-day period in 60 patients with cirrhosis and AKI (34 with AKI-HRS meeting the classical definition of type 1 HRS and 26 with AKI owing to acute tubular necrosis, AKI-ATN). Nineteen of 34 patients had resolution of AKI-HRS, while the remainder had persistent AKI-HRS. The persistence of AKI-HRS was associated with remarkably high short-term mortality. There were no significant differences in urinary NGAL or IL-18 between patients with resolution vs those with persistent AKI-HRS throughout the 14-day period. By contrast, biomarker levels were significantly lower in AKI-HRS, even if persistent, compared to AKI-ATN. These findings are highly suggestive of lack of significant tubular injury in AKI-HRS and could be of value in the clinical decision between combined liver-kidney or liver transplantation alone in patients with cirrhosis and AKI candidates to transplantation.

PD-L1 immunoexpression and molecular characterization of histological subtypes in urothelial carcinoma.

INTRODUCTION: Urothelial carcinoma (UC) has histological subtypes whose phenotype reflects their molecular diversity, behavior and response to conventional therapy. Immune checkpoint inhibitors (ICIs) have improved the management of UC by evaluation of PD-L1. In the case of PD-L1 22C3, the initiation of ICI is considered from a combined positive score (CPS) greater than 10. However, UC subtypes with absent PD-L1 22C3 expression in cases with CPS>10 may not respond to these treatments. This study aims to establish a correlation between the PD-L1 immunoexpression and molecular alterations in divergent differentiation and histological subtypes of UC (UC-s). MATERIAL AND METHODS: Twenty-six samples of UC were detected from a total of 24 patients. Two pathologists performed separately an assessment of UC-s on hematoxylin-eosin as well as PD-L1 expression. Molecular study of each case was performed by next generation sequencing (NGS). A descriptive analysis of the variables included was conducted. RESULTS: Nine cases (34.61%) showed a CPS>10, some with negative PD-L1 immunoexpression in aggressive UC-s. The molecular study revealed alterations in genes belonging to the p53/cell cycle control, RAS, and DNA repair pathways, among others. None of the alterations were exclusive to any histological subtype. DISCUSSION: Special attention should be paid to CPS>10 cases that include histological subtypes of UC with divergent expression for PD-L1 as they may not respond to treatment with ICI. We recommend examining the proportion and PD-L1 status of each subtype, especially if it has aggressive behavior.

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.

Three-Dimensional Anatomy of the Anterior Commissure: A Tractography and Anatomical Study.

BACKGROUND: The anterior commissure (AC) is one of the main commissural fibers of the brain. The commissural fibers are involved in bilateral integration and coordination of any normal brain activity. The AC is an important interhemispheric structure that forms a bidirectional communication channel between the frontal, temporal, parietal, and occipital lobes bilaterally. METHODS: In the present study, we focused on describing the morphology, relationships, and distribution of the AC using diffusion spectrum imaging-based fiber tracking. The tractograms were compared with the findings from gross anatomical dissection of the AC of adult brains. RESULTS: Our findings suggest that the AC found using tracking methods is larger than that found by dissection. CONCLUSIONS: The use of tractography added extensions to the main AC structure.

Rapid On-Farm Testing of Resistance in Lolium rigidum to Key Pre- and Post-Emergence Herbicides.

Overreliance on herbicides for weed control is conducive to the evolution of herbicide resistance. Lolium rigidum (annual ryegrass) is a species that is prone to evolve resistance to a wide range of herbicide modes of action. Rapid detection of herbicide-resistant weed populations in the field can aid farmers to optimize the use of effective herbicides for their control. The feasibility and utility of a rapid 7-d agar-based assay to reliably detect L. rigidum resistant to key pre- and post-emergence herbicides including clethodim, glyphosate, pyroxasulfone and trifluralin were investigated in three phases: correlation with traditional pot-based dose-response assays, effect of seed dormancy, and stability of herbicides in agar. Easy-to-interpret results were obtained using non-dormant seeds from susceptible and resistant populations, and resistance was detected similarly as pot-based assays. However, the test is not suitable for trifluralin because of instability in agar as measured over a 10-d period, as well as freshly-harvested seeds due to primary dormancy. This study demonstrates the utility of a portable and rapid assay that allows for on-farm testing of clethodim, glyphosate, and pyroxasulfone resistance in L. rigidum, thereby aiding the identification and implementation of effective herbicide control options.

Ocular surface characterization after allogeneic stem cell transplantation: A prospective study in a referral center.

Purpose: To characterize anatomical and functional changes in the ocular surface after hematopoietic stem cell transplantation. Methods: Three groups of patients were included in the study. Group 1: patients who had undergone allogeneic hematopoietic stem cell transplantation (HSCT) (n = 26). Group 2: patients who developed chronic graft versus host disease (GvHD) after HSCT (n = 14). Group 3: healthy subjects (n = 20). A complete ophthalmological examination was undertaken in all subjects, including Schirmer's test, TBUT (break-up-time) test, Oxford scale, OSDI test, corneal tomography, and conjunctival CD8+ lymphocyte detection. Results: In Branch 1 (comparative analysis before and after HSCT in Group 1), statistically significant differences were found in the following variables: best-corrected visual acuity (BCVA) OD (P = 0.08), OSDI test (P = 0.003), TBUT OU (OD P= 0, OS P= 0.0003), Oxford test OU (OD P= 0.01, OS P= 0.0049), and CD8+ lymphocytes OU (OD P= 0.003, OS P= 0.01). In Branch 2 (comparative analysis between Group 2 and 3), the variables with statistically significant differences (P < 0.001) in OU were: BCVA, OSDI test, Schirmer's test OU, TBUT test, Oxford test, and CD8+ lymphocytes. Finally, in Branch 3 (comparative analysis between Group 1 after HSCT and Group 2), statistically significant differences (P < 0.001) were found OU: in OSDI test, Schirmer's test, and Oxford test OU; and with P < 0.005 in TBUT test OU. Conclusion: In our study, statistically significant changes were observed in the OSDI test, TBUT test, Oxford Scale, and the detection of CD8+ lymphocytes in patients who underwent HSCT. Differences were more significant in those patients who had developed GvHD after HSCT compared to those without GvHD.

Psychological Burden of Hepatic Encephalopathy on Patients and Caregivers.

OBJECTIVES: Hepatic encephalopathy (HE) is common in advanced cirrhosis and is characterized by marked neuropsychiatric abnormalities. However, despite its severity and effects on brain function, the impact of HE on psychological status of patients has not been adequately assessed. The aim of this study was to evaluate the effect of HE on psychological status of patients and their informal caregivers. METHODS: Fifteen patients with cirrhosis and episodic or persistent HE and their corresponding informal caregivers were included. Semistructured interviews were performed in patients and caregivers. Quality of life (QoL) was assessed by the short-form 36 in both patients and caregivers, and the Zarit burden score was measured in caregivers. The analysis of interviews was performed using qualitative methodology. RESULTS: HE causes a major psychological impact on patients with HE. The first episode of HE caused a very significant impact that was reported with deep feelings, mainly of fear, anger, misery, anxiety, and sorrow, which persisted with time. Symptoms causing more psychological impact on patients were impaired ability to walk and speak. All effects were associated with a marked impairment in QoL. The psychological impact was also marked in caregivers who had a major burden, as assessed by the Zarit score. Moreover, QoL, particularly the mental component score, was markedly impaired in caregivers in intensity similar to that of patients. DISCUSSION: HE has a profound psychological impact on patients and their informal caregivers, associated with a marked negative influence on QoL. The psychological effects of HE on patients and caregivers should be evaluated and treated.

Características y manejo del cáncer de mama precoz en mujeres añosas asistidas en la Unidad Docente Asistencial de Mastología del Hospital de Clínicas en el período 2011-2018 / Characteristics and management of early breast cancer in elderly women assisted in the Mastology Care Teaching Unit of the Hospital de Clínicas in the period 2011-2018 / Características e manejo do câncer de mama precoce em idosas atendidas na Unidade de Ensino de Mastologia do Hospital de Clínicas no período de 2011-2018

Introducción: Existen pocas pautas para el tratamiento del cáncer de mama (CM) en pacientes añosas, lo que puede conducir al sub o sobre tratamiento. Objetivo: Conocer las características, manejo y la evolución del CM precoz en mujeres añosas. Material y métodos: Estudio observacional, descriptivo, transversal. Se recolectaron datos relacionados con las características clínico-patológicas y la evolución de pacientes de 70 años o más tratadas por CM en el período comprendido entre el 1/1/ 2011 y el 31/12/ 2018, asistidas en el Hospital de Clínicas. Se utilizaron herramientas de estadística descriptiva y para calcular la sobrevida global (SVG) se utilizó el método de Kaplan-Meier. Resultados: se incluyeron 31 pacientes; la edad mediana al diagnóstico fue 76,8 años; las características clínico-patológicas fueron: carcinoma ductal: 71%; GH 1-2: 74,2%; estadio I: 54,8 %; sin metástasis axilares: 80,6 %; HER2-RE/RP+ 80,6%; HER2+ 16,7%, y triple negativas 3,2%. El 29% de las pacientes fueron diagnosticadas mediante tamizaje poblacional y el 74,2% recibieron tratamiento según pautas vigentes, mientras que el 38,7% fueron subtratadas y el 16,1% sobretratadas. La mediana de SVG fue de 98,7 meses. Conclusiones: Una minoría de las pacientes fue diagnosticada mediante tamizaje poblacional, el tipo histológico más frecuente fue el ductal y la prevalencia de los tumores HER2-RE/RP+ fue mayor que en las pacientes más jóvenes. La mayoría de las pacientes recibió tratamiento estandar.

Introduction: There are few guidelines for the treatment of breast cancer (BC) in older patients, which can lead to under- or over-treatment. Objective: To understand the characteristics, management and evolution of early BC in older women. Material and methods: Observational, descriptive, cross-sectional study. Data were collected on the clinical-pathological characteristics and evolution of patients aged 70 years or older, treated for BC in the period from 1/1/ 2011 to 31/12/ 2018, at the Hospital de Clínicas. Descriptive statistical tools were used and the Kaplan-Meier method was applied to calculate the overall survival (OS) rate. Results: 31 patients were included; median age at diagnosis was 76.8 years old; the clinical-pathological characteristics were: ductal carcinoma: 71%; HG 1-2: 74.2%; stage I: 54.8%; no axillary metastases: 80.6%; HER2-ER/PR+ 80.6%; HER2+ 16.7%, and triple negative 3.2%. Of all the patients, 29% were diagnosed through screening and 74.2% were treated according to current guidelines, while 38.7% were under-treated and 16.1% over-treated. The median OS was 98.7 months. Conclusions: A minority of patients were diagnosed by screening, the most frequent histological type was ductal and the prevalence of HER2-RE/RP+ tumors was higher than in younger patients. Most patients received standard treatment.

Introdução: Existem poucas diretrizes para o tratamento do câncer de mama (CM) em pacientes idosos, o que pode levar ao sub ou excesso de tratamento. Objetivo: Conhecer as características, manejo e evolução do MC precoce em mulheres idosas. Material e métodos: estudo observacional, descritivo e transversal. Foram coletados dados relacionados às características clínico-patológicas e à evolução dos pacientes com 70 anos ou mais atendidos por CM no período de 01/01/2011 a 31/12/2018, atendidos no Hospital de Clínicas. Ferramentas de estatística descritiva foram usadas e o método de Kaplan-Meier foi usado para calcular a sobrevida global (SVG). Resultados: 31 pacientes foram incluídos; a mediana de idade ao diagnóstico foi de 76,8 anos; as características clínico-patológicas foram: carcinoma ductal: 71%; GH 1-2: 74,2%; estágio I: 54,8%; sem metástases axilares: 80,6%; HER2-RE / RP + 80,6%; HER2 + 16,7% e triplo negativo 3,2%. 29% dos pacientes foram diagnosticados por triagem populacional e 74,2% receberam tratamento de acordo com as diretrizes atuais, enquanto 38,7% foram subtratados e 16,1% supertratados. O SVG médio foi de 98,7 meses. Conclusões: A minoria dos pacientes foi diagnosticada por rastreamento populacional, o tipo histológico mais frequente foi ductal e a prevalência de tumores HER2-RE / RP + foi maior do que em pacientes mais jovens. A maioria dos pacientes recebeu tratamento padrão.

Incidence of cancer in a cohort of Spanish patients with systemic lupus erythematosus.

OBJECTIVE: To determine the incidence and prevalence of cancer in a cohort of patients with systemic lupus erythematosus (SLE) and identify associated risk factors. PATIENTS AND METHODS: The study comprised a dynamic cohort of SLE patients (November 1989 to December 2006) at a tertiary referral centre. An adjusted external control from the hospital tumour registry was used. RESULTS: The study included 175 SLE patients (90% women), with a mean time at risk of 1370.5 patient-years. Fourteen women (8%) died, mainly from cardiovascular events. No patient died due to malignancy. We found 35 tumours in 28 patients, 25 of them after the diagnosis of SLE, of which 5 were malignant. The rate of benign tumours was 14.6/1000 patient-years (95% CI, 8.9-22.5) and of malignant tumours 3.6/1000 patient-years (95% CI, 1.5-8.8), with a crude incidence odds ratio for malignant tumours of 3.5 (95% CI, 1.5-7.9). However, this significance was lost after standardizing the rates. Concerning associated risk factors, differences were found in the mean erythrocyte sedimentation rate [HR 1.4 (1.1-1.7)], and the presence of thrombosis [HR 6.9 (1.49-41.2)], especially arterial thrombosis. CONCLUSIONS: We found a crude incidence rate of cancer that was almost four times greater in our SLE patients as compared with the expected rate in the hospital area of western Malaga.

HPV Direct Flow CHIP: a new human papillomavirus genotyping method based on direct PCR from crude-cell extracts.

HPV Direct Flow CHIP is a newly developed test for identifying 18 high-risk and 18 low-risk human papillomavirus (HPV) genotypes. It is based on direct PCR from crude-cell extracts, automatic flow-through hybridization, and colorimetric detection. The aim of this study was to evaluate the performance of HPV Direct Flow CHIP in the analysis of 947 samples from routine cervical screening or the follow-up of abnormal Pap smears. The specimens were dry swab samples, liquid-based cytology samples, or formalin-fixed paraffin-embedded tissues. The genotype distribution was in agreement with known epidemiological data for the Spanish population. Three different subgroups of the samples were also tested by Linear Array (LA) HPV Genotyping Test (n=108), CLART HPV2 (n=82), or Digene Hybrid Capture 2 (HC2) HPV DNA Test (n=101). HPV positivity was 73.6% by HPV Direct Flow CHIP versus 67% by LA, 65.9% by HPV Direct Flow CHIP versus 59.8% by CLART, and 62.4% by HPV Direct Flow CHIP versus 42.6% by HC2. HPV Direct Flow CHIP showed a positive agreement of 88.6% with LA (k=0.798), 87.3% with CLART (k=0.818), and 68.2% with HC2 (k=0.618). In conclusion, HPV Direct Flow CHIP results were comparable with those of the other methods tested. Although further investigation is needed to compare the performance of this new test with a gold-standard reference method, these preliminary findings evidence the potential value of HPV Direct Flow CHIP in HPV vaccinology and epidemiology studies.

Monitoring respiration and biological stability during sludge composting with a modified dynamic respirometer.

A static electrolytic respirometer was adapted to work under dynamic conditions using an internal airflow recirculation system that passed air through sludge/straw solid mixtures. Airflow reduced oxygen transfer limitations and increased the maximum respiration rate and k(L)a values, indicating that the observed value may have been close to the actual biodegradation rate. Airflow caused sludge drying, so the sample moisture was controlled by air humidification. To apply the optimised respirometric technique, a pilotscale composting process was developed. Some commonly used respiration indices (RI(24), AT(4)) were used to measure the final compost stability. RI(24) indices without airflow were underestimated during the thermophilic composting stage. Once the easily biodegradable carbon was consumed, the static and dynamic RI(24) indices were nearly identical. Because of the dynamic procedure, the respiration rate was likely controlled by the biochemical reaction and not by the mass transfer. The respiration indices indicated that the final compost was unstable.