Imiquimod inhibits growth and induces differentiation of myeloid leukemia cell lines.

BACKGROUND: The antitumoral effects of different Toll-like receptor (TLRs) agonists is mediated by activating immune responses to suppress tumors growth, although TLR ligands may also have a direct effect on tumoral cells. Given that TLR signaling induces hematopoietic cell differentiations this may serve as a novel differentiation therapeutic approach for AML. METHODS: We investigated the effects of agonists for the ten human TLRs on the proliferation, apoptosis, cell cycle and differentiation of ten different types of myeloid leukemia cell lines (HL-60, U-937, KG-1, KG-1a, K-562, Kasumi-1, EOL-1, NB4, MOLM-13 and HEL). Proliferation was measured using the CellTiter 96® Aqueous One Solution Cell Proliferation Assay (Promega). Staining and analysis with a flow cytometer was used to identify cell cycle progression and apoptosis. Differentiation was measured by staining cells with the EuroFlow™ antibody panel for AML and analyzed by flow cytometry. FlowJo software was used to analyze the cytometric data. In all experiments, statistical significance was determined by a two-tailed t test. RESULTS: The activation of particular TLRs on some cell lines can induce growth inhibition and Imiquimod (a TLR 7 agonist) was the most effective agonist in all leukemic cell lines examined. Imiquimod was able to induce apoptosis, as well as to induce cell cycle alteration and upregulation of myeloid differentiation markers on some of the cell lines tested. CONCLUSIONS: Our results, together with the known efficacy of Imiquimod against many tumor entities, suggest that Imiquimod can be a potential alternative therapy to AML. This drug has a direct cytotoxic effect on leukemic cells, has the potential to induce differentiation, and can also stimulate the activation of cellular immune responses anti-AML.

PCR-Based Strategy for Introducing CRISPR/Cas9 Machinery into Hematopoietic Cell Lines.

Acute myeloid leukemia is a complex heterogeneous disease characterized by the clonal expansion of undifferentiated myeloid precursors. Due to the difficulty in the transfection of blood cells, several hematological models have recently been developed with CRISPR/Cas9, using viral vectors. In this study, we developed an alternative strategy in order to generate CRISPR constructs by fusion PCR, which any lab equipped with basic equipment can implement. Our PCR-generated constructs were easily introduced into hard-to-transfect leukemic cells, and their function was dually validated with the addition of MYBL2 and IDH2 genes into HEK293 cells. We then successfully modified the MYBL2 gene and introduced the R172 mutation into the IDH2 gene within NB4 and HL60 cells that constitutively expressed the Cas9 nuclease. The efficiency of mutation introduction with our methodology was similar to that of ribonucleoprotein strategies, and no off-target events were detected. Overall, our strategy represents a valid and intuitive alternative for introducing desired mutations into hard-to-transfect leukemic cells without viral transduction.

Predictive potential of SARS-CoV-2 RNA concentration in wastewater to assess the dynamics of COVID-19 clinical outcomes and infections.

Coronavirus disease 2019 - caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) -, has triggered a worldwide pandemic resulting in 665 million infections and over 6.5 million deaths as of December 15, 2022. The development of different epidemiological tools have helped predict new outbreaks and assess the behavior of clinical variables in different health contexts. In this study, we aimed to monitor concentrations of SARS-CoV-2 in wastewater as a tool to predict the progression of clinical variables during Waves 3, 5, and 6 of the pandemic in the Spanish city of Xátiva from September 2020 to March 2022. We estimated SARS-CoV-2 RNA concentrations in 195 wastewater samples using the RT-PCR Diagnostic Panel validated by the Center for Disease Control and Prevention. We also compared the trends of several clinical variables (14-day cumulative incidence, positive cases, hospital cases and stays, critical cases and stays, primary care visits, and deaths) for each study wave against wastewater SARS-CoV-2 RNA concentrations using Pearson's product-moment correlations, a two-sided Mann-Whitney U test, and a cross-correlation analysis. We found strong correlations between SARS-CoV-2 concentrations with 14-day cumulative incidence and positive cases over time. Wastewater RNA concentrations showed strong correlations with these variables one and two weeks in advance. There were significant correlations with hospitalizations and critical care during Wave 3 and Wave 6; cross-correlations were stronger for hospitalization stays one week before during Wave 6. No association between vaccination percentages and wastewater viral concentrations was observed. Our findings support wastewater SARS-CoV-2 concentrations as a potential surveillance tool to anticipate infection and epidemiological data such as 14-day cumulative incidence, hospitalizations, and critical care stays. Public health authorities could use this epidemiological tool on a similar population as an aid for health care decision-making during an epidemic outbreak.

Spanish version of the Heart Failure Somatic Perception Scale (HFSPS v.3) - psychometric properties.

Background: The Heart Failure Somatic Perception Scale (HFSPS) is an instrument that examine the existence and gravity of physical signs and symptoms in patients with heart failure, as well as early and subtle symptoms of HF that have clinical value, we aimed to translate and adapt the HFSPS from English to Spanish and evaluate the psychometric properties. Method: HFSPS translation and back translation were carried out according to the method established by of Beaton et al. A confirmatory factor analysis (CFA) was performed to test the factor structures. To assess criterion-related validity, HFSPS factor scores were correlated with Kansas City Cardiomyopathy Questionnaire (KCCQ) scores using the Spearman correlation method. The reliability of the internal consistency of the HFSPS was determined by calculating the Cronbach's alpha coefficient and the factor score determination coefficient. Results: Data from 173 patients with a mean age of 80.7 years (SD 9.1), women (51.1%), were analyzed. The majority (74.7%) were NYHA class II/III. The confirmatory factor analysis of four factors after eliminating one item showed fit indices close to the recommended indices: χ2 = 169.237, p < 0.001, CFI = 0.920, TLI = 0.901, RMSEA = 0.057 and SRMR = 0.061. Regarding the validity related to the criterion, all the scores of the HFSPS dimensions were correlated with all the scores of the KCCQ dimensions and were statistically significant. The reliability of the HFSPS factors of the coefficient of determination obtained scores of 0.73 for the dyspnea factor and early and subtle and lower for edema and chest discomfort with fewer items. Cronbach's alpha was acceptable for three of the scales >0.71 and poor 0.52 for chest discomfort with two items. The internal consistency index based on the model was 0.850. Conclusion: The Spanish version of the HFSPS is a valid and reliable instrument that that would be feasible to use in clinical and research setting to evaluate in the perception of symptoms in patients with heart failure.

Spanish Version of the Caregiver Contribution to Self-Care of Heart Failure Index (CC-SCHFI): A Psychometric Evaluation.

BACKGROUND: The Caregiver Contribution to Self-Care of Heart Failure (CC-SCHFI) is a theoretically driven instrument to measure the extent to which caregivers support heart failure (HF) patients to perform self-care. The CC-SCHFI measures caregivers' contribution to self-care maintenance and self-care management and caregiver confidence in contributing to heart failure patients' self-care. To date, the CC-SCHFI has never been tested in Spanish-speaking populations. PURPOSE: To translate the CC-SCHFI from English into Spanish and to test its psychometric characteristics. METHOD: CC-SCHFI translation and back-translation were performed according to the Beaton et al. methodology. Data from a cross-sectional study conducted in an outpatient clinic in Spain were used for the analysis. Psychometric analysis was performed with exploratory factor analysis (EFA) with oblique rotation. RESULTS: Caregivers had a mean age of 60.5 years (SD 14,9) and the majority were female (85%). Data from 220 caregivers were analyzed. From EFA, using the principal axis factoring method, we extracted two factors in the self-care maintenance subscale ("treatment adherence behaviors" and "symptom control and maintenance behaviors"), two in the self-care monitoring subscale ("illness behaviors" and "prevention behaviors") and one factor for the self-efficacy subscale. The Pearson's rank correlation coefficients between SCHFI and CCSCHFI showed significant correlation in each subdimension.

Case Report: Partial Uniparental Disomy Unmasks a Novel Recessive Mutation in the LYST Gene in a Patient With a Severe Phenotype of Chédiak-Higashi Syndrome.

Chédiak-Higashi syndrome (CHS) is a rare autosomal recessive (AR) immune disorder that has usually been associated to missense, nonsense or indels mutations in the LYST gene. In this study, we describe for the first time the case of a CHS patient carrying a homozygous mutation in the LYST gene inherited as a result of a partial uniparental isodisomy (UPiD) of maternal origin. Sanger sequencing of the LYST cDNA and single nucleotide polymorphism (SNP)-arrays were performed to identify the causative mutation and to explain the molecular mechanism of inheritance, respectively. Partial-UPiD leads to a copy neutral loss of heterozygosity (CN-LOH) of the telomeric region of chromosome 1 (1q41q44), unmasking the potential effect of the mutation detected. The mutation (c.8380dupT) is an insertion located in exon 32 of the LYST gene resulting in a premature stop codon and leading to the loss of all the conserved domains at the C-terminal of the LYST protein. This would account for the severe phenotype observed. We also reviewed the only two previously reported cases of CHS as a result of a uniparental disomy. In this study, we show that the combination of different strategies, including the use of SNP-arrays, is pivotal to fine-tune the diagnosis of rare AR disorders, such as CHS. Moreover, this case highlights the relevance of uniparental disomy as a potential mechanism of CHS expression in non-consanguineous families.

Transcultural Adaptation and Theoretical Models of Validation of the Spanish Version of the Self-Care of Heart Failure Index Version 6.2 (SCHFI v.6.2).

BACKGROUND: Heart failure (HF) is a major and growing public health problem worldwide. Across the world, heart failure is associated with high mortality, high hospitalization rates, and poor quality of life. Self-care is defined as a naturalistic decision-making process involving the choice of behaviors that maintain physiologic stability, the response to symptoms when they occur, and the ability to follow the treatment regimen and control symptoms. One instrument used to measure self-care is the Self Care of Heart Failure Index. AIM: The purpose of this study was to test the psychometric properties of the Spanish version of the Self Care of Heart Failure Index v.6.2 (SCHFI v.6.2). METHODOLOGY: Before testing its psychometric properties, the SCHFI v.6.2 was translated and adapted from its original English version into Spanish. Subsequently, we tested the instrument's psychometric properties on a sample of 203 participants with HF. Descriptive statistics were used to analyze the sociodemographic and clinical variables, and to describe item responses. We tested the factorial validity of the SCHFI v.6.2 using confirmatory and exploratory factor analysis. RESULTS: Confirmatory factor analysis (CFA) was performed using the our pre-existing models which resulted with poor fit indices. Thus, we performed exploratory factor analysis (EFA) on each of the SCHFI v.6.2 scales. CONCLUSION: The Spanish version of the SCHFI v.6.2. has good characteristics of factorial validity and can be used in clinical practice and research to measure self-care in patients with HF.

Qualitative Analysis by Experts of the Essential Elements of the Nursing Practice Environments Proposed by the TOP10 Questionnaire of Assessment of Environments in Primary Health Care.

Background: A short TOP10 scale based on the Practice Environment Scale-Nursing Work Index questionnaire measures the characteristics of nursing work environments. Positive environments result in better quality care and health outcomes. Objective: To identify a small number of core elements that would facilitate more effective interventions by nurse managers, and compare them with the essential elements proposed by the TOP10. Method: Qualitative research by a nominal group of eight experts. The content analysis was combined with descriptive data. Results: Ten most important items were selected and analyzed by the expert group. A high level of consensus in four items (2, 15, 20, 31) and an acceptable consensus in five items was reached (6, 11, 14, 18, 26). The tenth item in the top ten was selected from content analysis (19). The expert group agreed 90% with the elements selected as essential to the TOP10. Conclusion: The expert group achieved a high level of consensus that supports 90% of the essential elements of primary care settings proposed by the TOP10 questionnaire. Organizational changes implemented by managers to improve working environments must be prioritized following our results, so care delivery and health outcomes can be further improved.

Time and Cost of Hospitalisation for Salvage Therapy in Adults with Philadelphia Chromosome-Negative B Cell Precursor Relapsed or Refractory Acute Lymphoblastic Leukaemia in Spain.

BACKGROUND: Philadelphia chromosome-negative (Ph-) relapsed or refractory (R/R) B-cell precursor acute lymphoblastic leukaemia (ALL) is rare, and information on its impact on healthcare systems is scarce. OBJECTIVE: To quantify the time and reimbursement associated with hospitalisations of patients with R/R ALL in a Spanish hospital. METHODS: Retrospective review of medical charts identified patients aged ≥ 18 years with Ph- R/R ALL hospitalised between 1998 and 2014. Data were collected from the date of first diagnosis of R/R ALL (index) until death or loss to follow-up. The primary endpoint was the proportion of time hospitalised during chemotherapy. Reimbursement associated with hospitalisations (including associated chemotherapy) was also assessed. RESULTS: Thirty-two patients were eligible for inclusion. Their median age was 41 years, and 50% had a first remission duration of ≤ 1 year; 34% had undergone allogeneic haematological stem-cell transplantation (alloHSCT). Overall, 31 patients had received intensive salvage chemotherapy, during which there were 42 hospitalisations (mean 1.4/patient; mean duration 26 days). Patients spent a mean of 71% of the chemotherapy period in hospital. Total mean reimbursement was €26,417 per patient, almost all (€25,723) attributable to inpatient stays (€18,986/hospitalisation). From the index date to death or loss to follow-up (excluding alloHSCT-related hospitalisations), there were 80 hospitalisations (mean duration 24 days); mean reimbursement was €16,692 per hospitalisation and €41,730 per patient. AlloHSCT (n = 8) involved 18 hospitalisations (mean reimbursement €39,782/hospitalisation; €89,510/patient). CONCLUSION: Data from this sample of patients suggest that hospitalisations in R/R ALL are lengthy and associated with high costs in Spain.