Additional booster doses in patients with chronic lymphocytic leukemia induce humoral and cellular immune responses to SARS-CoV-2 similar to natural infection regardless ongoing treatments: A study by ERIC, the European Research Initiative on CLL.

Profound immune dysregulation and impaired response to the SARS-CoV-2 vaccine put patients with chronic lymphocytic leukemia (CLL) at risk of severe COVID-19. We compared humoral memory and T-cell responses after booster dose vaccination or breakthrough infection. (Green) Quantitative determination of anti-Spike specific antibodies. Booster doses increased seroconversion rate and antibody titers in all patient categories, ultimately generating humoral responses similar to those observed in the postinfection cohort. In detail, humoral response with overscale median antibody titers arose in >80% of patients in watch and wait, off-therapy in remission, or under treatment with venetoclax single-agent. Anti-CD20 antibodies and active treatment with BTK inhibitors (BTKi) represent limiting factors of humoral response, still memory mounted in ~40% of cases following booster doses or infection. (Blue) Evaluation of SARS-CoV-2-specific T-cell responses. Number of T-cell functional activation markers documented in each patient. The vast majority of patients, including those seronegative, developed T-cell responses, qualitatively similar between treatment groups or between vaccination alone and infection cases. These data highlight the efficacy of booster doses in eliciting T-cell immunity independently of treatment status and support the use of additional vaccination boosters to stimulate humoral immunity in patients on active CLL-directed treatments.

Generation of a bank of clinical-grade, HLA-homozygous iPSC lines with high coverage of the Spanish population.

BACKGROUND: Induced pluripotent stem cell (iPSC)-derived cell therapies are an interesting new area in the field of regenerative medicine. One of the approaches to decrease the costs of iPSC-derived therapies is the use of allogenic homozygous human leukocyte antigen (HLA)-matched donors to generate iPSC lines and to build a clinical-grade iPSC bank covering a high percentage of the Spanish population. METHODS: The Spanish Stem Cell Transplantation Registry was screened for cord blood units (CBUs) homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes. Seven donors were selected with haplotypes covering 21.37% of the haplotypes of the Spanish population. CD34-positive hematopoietic progenitors were isolated from the mononuclear cell fraction of frozen cord blood units from each donor by density gradient centrifugation and further by immune magnetic labeling and separation using purification columns. Purified CD34 + cells were reprogrammed to iPSCs by transduction with the CTS CytoTune-iPS 2.1 Sendai Reprogramming Kit. RESULTS: The iPSCs generated from the 7 donors were expanded, characterized, banked and registered. Master cell banks (MCBs) and working cell banks (WCBs) from the iPSCs of each donor were produced under GMP conditions in qualified clean rooms. CONCLUSIONS: Here, we present the first clinical-grade, iPSC haplobank in Spain made from CD34 + cells from seven cord blood units homozygous for the most common HLA-A, HLA-B and HLA-DRB1 haplotypes within the Spanish population. We describe their generation by transduction with Sendai viral vectors and their GMP-compliant expansion and banking. These haplolines will constitute starting materials for advanced therapy medicinal product development (ATMP).

Mucormycosis co-infection in COVID-19 patients: An update.

Mucormycosis (MCM) is a rare fungal disorder that has recently been increased in parallel with novel COVID-19 infection. MCM with COVID-19 is extremely lethal, particularly in immunocompromised individuals. The collection of available scientific information helps in the management of this co-infection, but still, the main question on COVID-19, whether it is occasional, participatory, concurrent, or coincidental needs to be addressed. Several case reports of these co-infections have been explained as causal associations, but the direct contribution in immunocompromised individuals remains to be explored completely. This review aims to provide an update that serves as a guide for the diagnosis and treatment of MCM patients' co-infection with COVID-19. The initial report has suggested that COVID-19 patients might be susceptible to developing invasive fungal infections by different species, including MCM as a co-infection. In spite of this, co-infection has been explored only in severe cases with common triangles: diabetes, diabetes ketoacidosis, and corticosteroids. Pathogenic mechanisms in the aggressiveness of MCM infection involves the reduction of phagocytic activity, attainable quantities of ferritin attributed with transferrin in diabetic ketoacidosis, and fungal heme oxygenase, which enhances iron absorption for its metabolism. Therefore, severe COVID-19 cases are associated with increased risk factors of invasive fungal co-infections. In addition, COVID-19 infection leads to reduction in cluster of differentiation, especially CD4+ and CD8+ T cell counts, which may be highly implicated in fungal co-infections. Thus, the progress in MCM management is dependent on a different strategy, including reduction or stopping of implicit predisposing factors, early intake of active antifungal drugs at appropriate doses, and complete elimination via surgical debridement of infected tissues.

Parkinson's disease patient-specific neuronal networks carrying the LRRK2 G2019S mutation unveil early functional alterations that predate neurodegeneration.

A deeper understanding of early disease mechanisms occurring in Parkinson's disease (PD) is needed to reveal restorative targets. Here we report that human induced pluripotent stem cell (iPSC)-derived dopaminergic neurons (DAn) obtained from healthy individuals or patients harboring LRRK2 PD-causing mutation can create highly complex networks with evident signs of functional maturation over time. Compared to control neuronal networks, LRRK2 PD patients' networks displayed an elevated bursting behavior, in the absence of neurodegeneration. By combining functional calcium imaging, biophysical modeling, and DAn-lineage tracing, we found a decrease in DAn neurite density that triggered overall functional alterations in PD neuronal networks. Our data implicate early dysfunction as a prime focus that may contribute to the initiation of downstream degenerative pathways preceding DAn loss in PD, highlighting a potential window of opportunity for pre-symptomatic assessment of chronic degenerative diseases.

Vitamin D receptor (VDR) expression in different molecular subtypes of canine mammary carcinoma.

BACKGROUND: The molecular-based classification of canine mammary carcinomas (CMCs) has been the focus of much current research. Both in canines and humans, the triple-negative (TN) molecular subtype of mammary cancer is defined by a lack of expression of progesterone receptor (PR), oestrogen receptor (ER) and HER2. It has a poor prognosis; no effective targeted therapy is available. Vitamin D displays anticarcinogenic properties, and the expression of its receptor (VDR) has been found in different molecular subtypes, being about 30-40 % of TN breast cancer (TNBC) positive to it. We assessed the VDR expression in the different molecular subtypes of 58 CMCs from 45 female dogs using an immunohistochemical panel for the molecular classification of included: PR, ER, HER2, cytokeratin (CK) 5, CK14, and Ki67. In addition, we studied the relationship among the molecular subtypes of CMCs and clinicopathologic parameters. RESULTS: Investigation showed VDR positivity in 45.0 % of the triple-negative CMCs (TNCMCs), 27.3 % of luminal B and 19.0 % of luminal A. Luminal A was the most molecular subtype represented of the total tumours (36.2 %), followed of TNCMCs (34.5 %), luminal B (20.7 %) and HER2-overexpression (10.3 %). Both HER2-overexpression and TNCMC subtypes were positively related to lymphatic invasion (P = 0.028), simple histologic subtype (P = 0.007), a higher histological grade (P = 0.045) and a trend to higher proliferation index (P = 0.09). CONCLUSIONS: The highest VDR expression was observed in TNCMC, being almost half of them (45 %) positive to this receptor. VDR expression was absent in HER2-overexpression tumours and low in luminal A and B molecular subtypes.

Patient-specific iPSC-derived cellular models of LGMDR1.

Limb-girdle muscular dystrophy recessive 1 (LGMDR1) represents one of the most common types of LGMD in the population, where patients develop a progressive muscle degeneration. The disease is caused by mutations in calpain 3 gene, with over 500 mutations reported to date. However, the molecular events that lead to muscle wasting are not clear, nor the reasons for the great clinical variability among patients, and this has so far hindered the development of effective therapies. Here we generate human induced pluripotent stem cells (iPSCs) from skin fibroblasts of 2 healthy controls and 4 LGMDR1 patients with different mutations. The generated lines were able to differentiate into myogenic progenitors and myotubes in vitro and in vivo, upon a transient PAX7 overexpressing protocol. Thus, we have generated myogenic cellular models of LGMDR1 that harbor different CAPN3 mutations within a human genetic background, and which do not derive from muscular biopsies. These models will allow us to investigate disease mechanisms and test therapies. Despite the variability found among iPSC lines that was unrelated to CAPN3 mutations, we found that patient-derived myogenic progenitors and myotubes express lower levels of DMD, which codes a key protein in satellite cell regulation and myotube maturation.

Abicipar pegol for neovascular age-related macular degeneration.

INTRODUCTION: The development of intravitreal anti-vascular endothelial growth factor (VEGF) therapy has revolutionized management of neovascular age-related macular degeneration (nAMD) and serves as the standard of care for treating this chronic, progressive disease. One shortcoming is the need for frequent intravitreal injections to maintain visual gains, which has led to pursuit of long-acting agents to reduce treatment burden. AREAS COVERED: A literature search was conducted using the keywords 'abicipar pegol' and 'DARPin' on PubMed. EXPERT OPINION: DARPin (Designed Ankyrin Repeat Proteins) molecules such as abicipar pegol offer potential therapeutic advantages over antibodies or antibody fragments, including high affinity, stability, and high molar concentration. The phase III SEQUOIA and CEDAR clinical trials suggest that abicipar allows >90% of patients to maintain stable vision with 12-week dosing intervals, comparable to results achieved with monthly ranibizumab injections. Relative to other anti-VEGF agents, intraocular inflammation has been noted in a concerning percentage of patients, which is hypothesized to be related to the manufacturing process rather than the drug itself. Modifications to reduce pro-inflammatory components resulted in reduced inflammation (8.9%) in the MAPLE study. If this high inflammation rate can be further reduced, abicipar has the potential to decrease treatment burden for nAMD patients.

How Can We Improve the Selection of Patients Awaiting Liver Retransplantation?

OBJECTIVE: To analyze predictors of survival involved in liver retransplantation (LRT), including the Rosen Model (RM). MATERIALS AND METHODS: This was a descriptive, observational, and unicentric study based on predictors of survival including patients who underwent LRT in a tertiary medical center between April 2002 and December 2018. Recipient, donor, and transplant data were collected, and RM score was calculated for every patient. Fisher exact test and Student t test were used for qualitative and quantitative variables, respectively. The Shapiro-Wilks test was applied to verify the normality of the sample. Survival differences between subgroups were checked using the log-rank test. Statistical significance was stated at P < .05. RESULTS: Among 32 retransplanted patients in this period, 17 (53.1%) survived more than 12 months after LRT. The results of statistical associations between prognostic factors and overall survival highlighted that an older recipient age was significantly correlated with a lower overall survival. The 3-month overall survival was 84.3%. Nineteen patients had a low risk according to RM, with a 3-month survival rate of 78.9%. Eight had a RM intermediate risk, with a survival rate of 21%. Despite the aforementioned data, the log-rank test did not find statistical differences in survival (P = .488). CONCLUSION: We should consider older recipient age as a negative prognostic factor of overall survival. Also, we should contemplate intermediate risk according to RM as an adverse predictor regarding survival in LRT. Both data are of interest regarding the indication or not of LRT and prioritization on the waiting list.

Liver Retransplantation: The Changing Scenario in a Tertiary Medical Center.

AIM: To analyze the causes of liver retransplantation (LRT), which mostly depend on recipient factors. MATERIALS AND METHODS: A descriptive, observational, and unicentric study including patients who underwent an LRT in a tertiary medical center between April 2002 and December 2018. Recipient, donor, and liver transplant data were collected. RESULTS: During the period under review a total of 468 transplants were made; among them, 32 (6.8%) were LRT. The most common indication (25%) was hepatic artery thrombosis (HAT) developing ischemic cholangiopathy followed by chronic rejection (21.8%). Late LRT was performed in 71.8%. A total of 96.8% of donations were after brain death with a donor median age of 65 years. Six patients (18.7%) had HAT as a postoperative complication. The recipients' 3-, 6-, and 12-month overall survival was 72.7%, 54.6%, and 51.5%, respectively, and the 5-year was 46.8%. Leading cause of death was septic shock (42.1%). CONCLUSION: In our patients, the most common cause of LRT is HAT. We had an LRT rate of 6.8%, which is consistent with national and international registers.

Production of insulin-like growth factor-binding proteins during the development of hepatic fibrosis due to chronic hepatitis C. / Producción de las proteínas de unión al factor de crecimiento insulinoide durante el desarrollo de la fibrosis hepática por hepatitis C crónica.

INTRODUCTION AND AIMS: Insulin-like growth factor 1 is modulated by the insulin-like growth factor-binding proteins (IGFBPs) that are synthesized in the liver. The aim of the present study was to evaluate the concentrations of IGFBPs 1-7 in patients with chronic hepatitis C and study their association with fibrosis stage. PATIENTS AND METHODS: A prospective, cross-sectional study was conducted that included patients with chronic hepatitis C. The stages of fibrosis were determined through FibroTest and FibroScan and the patients were compared with a control group. Serum levels of IGFBPs 1-7 were quantified through multiple suspension arrays. The Kruskal-Wallis test, Mann-Whitney U test, Spearman's correlation, and ROC curves were used for the statistical analysis. RESULTS: Upon comparing the patients and controls, the highest concentrations were found in IGFBPs 1, 2, 4, and 7 (p=0.02, p=0.002, p=0.008, and p<0.001, respectively). IGFBP-3 levels had a tendency to be lower in the patients (p=0.066), whereas values were similar between patients and controls for IGFBP-5 and 6 (p=0.786 and p=0.244, respectively). Of the seven IGFBPs, IGFBP-3 concentrations were the highest. There were significant differences between fibrosis stages for IGFBP-5 and IGFBP-7. CONCLUSION: IGFBPs play a relevant role in the fibrotic process in liver damage. IGFBP-7, in particular, differentiates fibrosis stages, making it a potential serum biomarker.

Barut-Girardello coherent states for anisotropic 2D-Dirac materials.

We construct the Barut-Girardello coherent states for charge carriers in anisotropic 2D-Dirac materials immersed in a constant homogeneous magnetic field which is orthogonal to the sample surface. For that purpose, we solve the anisotropic Dirac equation and identify the appropriate arising and lowering operators. Working in a Landau-like gauge, we explicitly construct nonlinear coherent states as eigenstates of a generalized annihilation operator with complex eigenvalues which depends on an arbitrary function f  of the number operator. In order to describe the anisotropy effects on these states, we obtain the Heisenberg uncertainty relation, the probability density, mean energy value and occupation number distribution for three different functions f . For the case in which the anisotropy is caused by uniaxial strain, we obtain that when the stress is applied along the x-axis of the material surface, the probability density for the nonlinear coherent states is smaller compared to when the material is stressed along the orthogonal axis.

Influence of Donor Obesity on Long-Term Liver Transplantation Outcomes.

OBJECTIVE: To analyze liver transplantation outcomes according to the body mass index (BMI) of donors. MATERIAL AND METHODS: A retrospective descriptive study was conducted in patients transplanted at our center between January 2006 and December 2014, comparing outcomes between grafts from obese (body mass index [BMI] ≥30) and nonobese (BMI ≤30) donors. We analyzed the reasons for transplantation, the morbidity-mortality related variables, and survival after a minimum follow-up of 24 months. A multivariate logistic model was constructed to predict the mortality. Survival was analyzed with the Kaplan-Meier method, and survival curves were compared using the log-rank test. RESULTS: The study included 50 obese and 175 nonobese donors. A significant difference between the groups was found in the pre-extraction intensive care unit (ICU) stay of the donors (P = .006) but not in the post-transplantation complications or survival of the respective recipients (P > .05). In the multivariate analysis, mortality was significantly associated with the presence of the hepatitis C virus (HCV) (P = .001) in the recipient and with the age of the donor (P = .043), finding the risk of death to be 2.87-fold higher in patients with HCV versus without HCV (95% confidence interval [1.641-5.043]) and 1.7% higher with every additional year of donor age (odds ratio 1.017, 95% confidence interval [1-1.034]). CONCLUSIONS: A significantly longer pre-extraction ICU stay was observed in obese (BMI ≥30) versus nonobese (BMI <30) donors, but no significant between-group difference was found in the post-transplant complications or survival of the respective recipients. The mortality risk was higher in HCV-positive recipients and in those receiving grafts from older donors.

Liver Transplantation in Recipients With High Model for End-stage Liver Disease Score.

OBJECTIVES: The score in the Model of End-stage Liver Disease, or MELD, is a good indicator of the survival in patients on the liver transplant waiting list. In this study, an analysis is performed on the benefits of liver transplant on those patients with a very high MELD score and who thus start from a very severe baseline state that could affect the surgical outcome. MATERIALS AND METHODS: A prospective study was conducted on a cohort of 331 patients that received a liver transplant between 2002 and 2014. The patients were divided into 2 groups according to the MELD score (<28 vs ≥28), and differences in age, postoperative complications, stay in the intensive care unit (ICU), hospital stay, and survival were compared. RESULTS: Of the total of 331 patients, 21 (6.3%) had a MELD score ≥ 28. The mean age of the group with MELD score ≥ 28 was lower than the age in the group with MEDL score < 28 (42.5 vs 53.7 years; P < .0001). No significant increase was observed in postoperative complications. Although there were also no differences in survival, the group with MELD score ≥ 28 did have a longer stay in ICU and a longer hospital stay (with a mean of 6.7 days in ICU and 41.5 days admission vs 4.1 and 26.9, respectively). CONCLUSIONS: A very high MELD score is associated with a longer stay in ICU and more days of hospital admission, although no differences were observed in postoperative complications or survival. Therefore, there does not seem to be any contraindication in transplantation in this group of patients.

Analysis of Ischemic Cholangiopathy After Treatment of Arterial Thrombosis in Liver Transplantation in Our Series.

Liver transplantation offers patients with terminal liver disease an effective treatment accompanied by excellent quality of life, but it also has complications, such as hepatic artery thrombosis and development of ischemic cholangiopathy, described in 3%-17% of patients. It is a very important cause of morbidity and mortality. The objective of this report was to analyze the efficacy of the treatment they received in relation to the development of ischemic cholangiopathy and a comparative survival analysis and to propose prophylactic measures for high-risk patients.

Retracted: Methotrexate in the treatment of symptomatic knee osteoarthritis: randomised placebo-controlled trial.

This article has been retracted.

Vitamin D receptor expression in canine mammary gland and relationship with clinicopathological parameters and progesterone/oestrogen receptors.

The vitamin D receptor (VDR) belongs to the nuclear class II receptor family. VDR is a ligand transcription factor and mediates the actions of calcitriol, the active product of vitamin D synthesis. Nowadays, it is known that the biological actions of calcitriol include the capacity to modulate cancer features, such as proliferation and differentiation, apoptosis, angiogenesis, invasion and metastasis. VDR expression has been demonstrated in human breast cancer and vitamin D has emerged as a promising targeted therapy. We analyse the VDR expression in normal and neoplastic canine mammary tissue samples and its relationship with clinicopathological parameters and progesterone/oestrogens receptors (PR/ER). Expression of VDR, Ki67 (to evaluate the proliferation index, PI), PR and ER was assessed in 50 mammary gland tissue samples from 41 female dogs by immunohistochemistry. VDR-positive staining was found in the nuclei of both myoepithelial and luminal epithelial cell layers. VDR expression was higher in normal mammary tissue (37/37 cases, 100%) then followed by benign tumours (6/15 cases, 40%) and malignant tumours (9/34 cases, 26.5%) (P = .001). Female dogs aged ≥10 years had lower VDR expression compared with dogs younger (P = .017). Relationship between VDR and breed, number of tumours, tumour size, histologic subtype, histologic grade of malignancy, PI and PR and ER expression was not observed. Studies with more samples are necessary to further evaluate the possible role of VDR in the biological behaviour of canine mammary tumours, and to corroborate the possibility to use the dog as model for human breast cancer.

Generation of six multiple sclerosis patient-derived induced pluripotent stem cell lines.

Multiple sclerosis (MS) is considered a chronic autoimmune disease of the central nervous system that leads to gliosis, demyelination, axonal damage and neuronal death. The MS disease aetiology is unknown, though a polymorphism of the TNFRSF1A gene, rs1800693, is known to confer an increased risk for MS. Using retroviral delivery of reprogramming transgenes, we generated six MS patient-specific iPSC lines with two distinct genotypes, CC or TT, of the polymorphism rs1800693. iPSC lines had normal karyotype, expressed pluripotency genes and differentiated into the three germ layers. These lines offer a good tool to study MS pathomechanisms and for drug testing.

Biomarker responses of Cu-induced toxicity in European seabass Dicentrarchus labrax: Assessing oxidative stress and histopathological alterations.

A comprehensive approach to chemical accumulation and biological effects of short-term Cu exposure in juveniles of European seabass (Dicentrarchus labrax) has been achieved. Fish were exposed to 0.01-10mgL-1 nominal Cu concentrations for 24-96h. Metal concentrations in water and gills, liver, muscle and brain tissues were studied along with oxidative stress biomarkers (superoxide dismutase, catalase, glutathione peroxidase, lipid peroxidation). Induction of oxidative damage was observed in all the organs with differential antioxidant responses; gills appearing as the most sensitive from low environmentally water Cu concentrations as 0.01mgL-1. Histopathological alterations were also observed in liver and gills, even without a significant Cu accumulation. The results show that the combination of oxidative stress parameters, particularly lipid peroxidation and glutathione peroxidase activities, and histopathological alterations provide a good model fish and reliable early biomarkers for monitoring Cu pollution in seawater and might call for the protection agencies to revise the Cu environmental standards.

Advanced cell-based modeling of the royal disease: characterization of the mutated F9 mRNA.

Essentials The Royal disease (RD) is a form of hemophilia B predicted to be caused by a splicing mutation. We generated an iPSC-based model of the disease allowing mechanistic studies at the RNA level. F9 mRNA analysis in iPSC-derived hepatocyte-like cells showed the predicted abnormal splicing. Mutated F9 mRNA level was very low but we also found traces of wild type transcripts. SUMMARY: Background The royal disease is a form of hemophilia B (HB) that affected many descendants of Queen Victoria in the 19th and 20th centuries. It was found to be caused by the mutation F9 c.278-3A>G. Objective To generate a physiological cell model of the disease and to study F9 expression at the RNA level. Methods Using fibroblasts from skin biopsies of a previously identified hemophilic patient bearing the F9 c.278-3A>G mutation and his mother, we generated induced pluripotent stem cells (iPSCs). Both the patient's and mother's iPSCs were differentiated into hepatocyte-like cells (HLCs) and their F9 mRNA was analyzed using next-generation sequencing (NGS). Results and Conclusion We demonstrated the previously predicted aberrant splicing of the F9 transcript as a result of an intronic nucleotide substitution leading to a frameshift and the generation of a premature termination codon (PTC). The F9 mRNA level in the patient's HLCs was significantly reduced compared with that of his mother, suggesting that mutated transcripts undergo nonsense-mediated decay (NMD), a cellular mechanism that degrades PTC-containing mRNAs. We also detected small proportions of correctly spliced transcripts in the patient's HLCs, which, combined with genetic variability in splicing and NMD machineries, could partially explain some clinical variability among affected members of the European royal families who had lifespans above the average. This work allowed the demonstration of the pathologic consequences of an intronic mutation in the F9 gene and represents the first bona fide cellular model of HB allowing the study of rare mutations at the RNA level.