SARS-CoV-2 Infection and Anemia-A Focus on RBC Deformability and Membrane Proteomics-Integrated Observational Prospective Study.

INTRODUCTION: The multifaceted impact of COVID-19 extends beyond the respiratory system, encompassing intricate interactions with various physiological systems. This study elucidates the potential association between SARS-CoV-2 infection and anemia, with a particular emphasis on the deformability of red blood cells (RBCs), stability of hemoglobin, enzymatic activities, and proteomic profiles. METHODS: The study encompasses a cohort of 74 individuals, including individuals positive for COVID-19, a control group, and patients with other viral infections to discern the specific effects attributable to COVID-19. The analysis of red blood cells was focused on deformability measured by osmotic gradient ektacytometry, hemoglobin stability, and glycolytic enzyme activity. Furthermore, membrane proteins were examined using advanced proteomics techniques to capture molecular-level changes. RESULTS: Findings from the study suggest a correlation between anemia and exacerbated outcomes in COVID-19 patients, marked by significant elevations in d-dimer, serum procalcitonin, creatinine, and blood urea nitrogen (BUN) levels. These observations suggest that chronic kidney disease (CKD) may play a role in the development of anemia in COVID-19 patients, particularly those of advanced age with comorbidities. Furthermore, the proteomic analyses have highlighted a complex relationship between omics data and RBC parameters, enriching our understanding of the mechanisms underlying the disease. CONCLUSIONS: This research substantiates the complex interrelationship between COVID-19 and anemia, with a specific emphasis on the potential repercussions of SARS-CoV-2 infection on RBCs. The findings contribute to the growing body of evidence supporting the extensive impact of COVID-19 on RBCs.

Sample preparation for suspect screening of persistent, mobile and toxic substances and their phase II metabolites in human urine by mixed-mode liquid chromatography.

Persistent, mobile and toxic substances have drawn attention nowadays due to their particular properties, but they are overlooked in human monitorization works, limiting the knowledge of the human exposome. In that sense, human urine is an interesting matrix since not only parent compounds are eliminated, but also their phase II metabolites that could act as biomarkers. In this work, 11 sample preparation procedures involving preconcentration were tested to ensure maximum analytical coverage in human urine using mixed-mode liquid chromatography coupled with high-resolution tandem mass spectrometry. The optimized procedure consisted of a combination of solid-phase extraction and salt-assisted liquid-liquid extraction and it was employed for suspect screening. Additionally, a non-discriminatory dilute-and-shoot approach was also evaluated. After evaluating the workflow in terms of limits of identification and type II errors (i.e., false negatives), a pooled urine sample was analysed. From a list of 1450 suspects and in-silico simulated 1568 phase II metabolites (i.e. sulphates, glucuronides, and glycines), 44 and 14 substances were annotated, respectively. Most of the screened suspects were diverse industrial chemicals, but biocides, natural products and pharmaceuticals were also detected. Lastly, the complementarity of the sample preparation procedures, columns, and analysis conditions was assessed. As a result, dilute-and-shoot and the Acclaim Trinity P1 column at pH = 3 (positive ionization) and pH = 7 (negative ionization) allowed the maximum coverage since almost 70 % of the total suspects could be screened using those conditions.

Development and evaluation of a comprehensive workflow for suspect screening of exposome-related xenobiotics and phase II metabolites in diverse human biofluids.

Suspect and non-target screening (SNTS) methods are being promoted in order to decode the human exposome since a wide chemical space can be analysed in a diversity of human biofluids. However, SNTS approaches in the exposomics field are infra-studied in comparison to environmental or food monitoring studies. In this work, a comprehensive suspect screening workflow was developed to annotate exposome-related xenobiotics and phase II metabolites in diverse human biofluids. Precisely, human urine, breast milk, saliva and ovarian follicular fluid were employed as samples and analysed by means of ultra-high performance liquid chromatography coupled with high resolution tandem mass spectrometry (UHPLC-HRMS/MS). To automate the workflow, the "peak rating" parameter implemented in Compound Discoverer 3.3.2 was optimized to avoid time-consuming manual revision of chromatographic peaks. In addition, the presence of endogenous molecules that might interfere with the annotation of xenobiotics was carefully studied as the employment of inclusion and exclusion suspect lists. To evaluate the workflow, limits of identification (LOIs) and type I and II errors (i.e., false positives and negatives, respectively) were calculated in both standard solutions and spiked biofluids using 161 xenobiotics and 22 metabolites. For 80.3 % of the suspects, LOIs below 15 ng/mL were achieved. In terms of type I errors, only two cases were identified in standards and spiked samples. Regarding type II errors, the 7.7 % errors accounted in standards increased to 17.4 % in real samples. Lastly, the use of an inclusion list for endogens was favoured since it avoided 18.7 % of potential type I errors, while the exclusion list caused 7.2 % of type II errors despite making the annotation workflow less time-consuming.

Diagnosis and clinical management of thrombotic thrombocytopenic purpura (TTP): a consensus statement from the TTP Catalan group.

Thrombotic thrombocytopenic purpura (TTP) is a low prevalence disease characterized by severe deficiency of the enzyme ADAMTS13, leading to the development of thrombotic microangiopathy (TMA) and often resulting in severe organ disfunction. TTP is an extremely serious condition and, therefore, timely and appropriate treatment is critical to prevent life-threatening complications.Over the past 25 years, significant advances in the understanding of the pathophysiology of immune TTP have led to the development of readily available techniques for measuring ADAMTS13 levels, as well as new drugs that are particularly effective in the acute phase and in preventing relapses. These developments have improved the course of the disease.Given the complexity of the disease and its various clinical and laboratory manifestations, early diagnosis and treatment can be challenging.To address this challenge, a group of experienced professionals from the Catalan TTP group have developed this consensus statement to standardize terminology, diagnosis, treatment and follow up for immune TTP, based on currently available scientific evidence in the field. This guidance document aims to provide healthcare professionals with a comprehensive tool to make more accurate and timely diagnosis of TTP and improve patient outcomes.

José Martí en las artes visuales como motivación en la asignatura Promoción de Salud

Introducción: José Martí es un ejemplo de hombre de formación esencialmente humanística, que percibió a plenitud la necesidad de poseer una cultura general integral. Objetivo: describir la importancia del uso de la figura de José Martí en las artes visuales, como herramienta de motivación para los estudiantes del primer año de Medicina, en la asignatura Promoción de Salud. Desarrollo: la educación se defiende con el compromiso, la creatividad y el ejemplo, trabajando por la belleza y la cultura del detalle, como práctica en cada centro educacional. Teniendo en cuenta los contenidos y los objetivos educativos de la asignatura Promoción de Salud, ha sido muy positivo utilizar la figura de Martí, vista por diferentes artistas visuales, como elemento motivador para impartir las diferentes actividades docentes de la asignatura Promoción de Salud. Conclusiones: las acciones permiten ayudar a la formación de valores éticos y estéticos de los estudiantes del primer año de Medicina de la Universidad de Ciencias Médicas de Matanzas, y lograr en ellos una cultura general integral.

Introduction: Jose Marti is an example of a man of essentially humanistic formation, who fully perceived the need of having a comprehensive general culture. Objective: to describe the importance of the use of the figure of Jose Marti in the visual arts, as a motivational tool for first-year Medicine students, in the subject Health Promotion. Development: education is defended with commitment, creativity and example, working for the beauty and culture of detail, as a practice in each educational center. Taking into account the contents and educational objectives of the subject Health Promotion, it has been very positive to use the figure of Marti, seen by different artists of the visual arts, to impart the different teaching activities of the subject Health Promotion. Conclusions: the actions allow to help the formation of ethical and esthetic values of the first-year Medicine students of the Matanzas University of Medical Sciences, and to achieve in them a comprehensive general culture.

Desafíos para los médicos de familia ante la pandemia de COVID-19 / Challenges for family doctors in the face of the COVID-19 pandemic

RESUMEN La atención primaria de salud es el área de actuación fundamental de la medicina familiar y una herramienta clave para su desarrollo. Al ser declarada como pandemia por la Organización Mundial de la Salud, la COVID-19 ha hecho que los sistemas de salud consideren los sólidos servicios de atención primaria como base de la respuesta ante esta emergencia. El objetivo del presente trabajo fue describir los desafíos a los que se enfrentan los médicos de familia en la atención a pacientes sospechosos o confirmados por la COVID-19. Es necesario que el personal de salud y toda la población conozcan las características del nuevo coronavirus. En Cuba, existe un Protocolo Nacional contra la COVID-19, en constante actualización, donde se definen los criterios de casos, las principales medidas preventivas y el tratamiento para combatir la enfermedad. El médico familiar debe establecer consultas de clasificación para atender los pacientes con enfermedades respiratorias agudas; de esta manera se evita el contacto con los demás pacientes que acuden con otros problemas de salud. Asimismo, debe insistir en las medidas preventivas y tratar los casos sospechosos y confirmados, mantenerse actualizado, realizar pesquisa activa de los casos, garantizar la atención de las personas que se encuentran en los centros de aislamiento, e investigar cada una de las características de la enfermedad. A modo de conclusión, se plantea que la medicina familiar es necesaria para prevenir la enfermedad por coronavirus 2019. Son varios los retos y desafíos que deben enfrentar los médicos de familia ante la pandemia (AU).

ABSTRACT Primary health care is the main area of action of family medicine and a key tool for its development. Being declared a pandemic by the World Health Organization, COVID-19 has made health systems consider solid primary care services as the basis of the response to this emergency. The aim of this paper was to describe the challenges faced by family physicians in taking care of COVID-19 suspected or confirmed patients. The health care staff and the entire population need to know the characteristics of the novel coronavirus. There is a National Protocol against COVID-19 in Cuba that is regularly updated; it defines the cases criteria, the main preventive measures and the treatment to fight the disease. The family doctor should establish classification consultations to take care of patients with acute respiratory diseases; this avoids contact with the rest of patients who come with other health problems. They should also insist on preventive measures and take care of suspicious and confirmed cases; keep themselves updated; carry out an active investigation of cases; ensure the care of persons in isolation centers, and study each of the characteristics of the disease. As a conclusion, it is indicated that family medicine is necessary to prevent coronavirus disease 2019. There are several challenges and defies that family doctors must face in the face of the pandemic (AU).

Cellulose biosynthesis using simple sugars available in residual cacao mucilage exudate.

Worldwide only 8% of the biomass from harvested cacao fruits is used, as cacao beans, in chocolate-based products. Cacao mucilage exudate (CME), a nutrient-rich fluid, is usually lost during cacao beans fermentation. CME's composition and availability suggest a potential carbon source for cellulose production. CME and the Hestrin and Schramm medium were used, and compared, as growth media for bacterial cellulose (BC) production with Gluconacetobacter xylinus. CME can be used to produce BC. However, the high sugar content, low pH, and limited nitrogen sources in CME hinder G. xylinus growth affecting cellulose yields. BC production increased from 0.55 ± 0.16 g L-1 up to 13.13 ± 1.09 g L-1 after CME dilution and addition of a nitrogen source. BC production was scaled up from 30 mL to 15 L, using lab-scale experiments conditions, with no significant changes in yields and production rates, suggesting a robust process with industrial possibilities.

Genome sequencing and functional characterization of a Dictyopanus pusillus fungal enzymatic extract offers a promising alternative for lignocellulose pretreatment of oil palm residues.

The pretreatment of biomass remains a critical requirement for bio-renewable fuel production from lignocellulose. Although current processes primarily involve chemical and physical approaches, the biological breakdown of lignin using enzymes and microorganisms is quickly becoming an interesting eco-friendly alternative to classical processes. As a result, bioprospection of wild fungi from naturally occurring lignin-rich sources remains a suitable method to uncover and isolate new species exhibiting ligninolytic activity. In this study, wild species of white rot fungi were collected from Colombian forests based on their natural wood decay ability and high capacity to secrete oxidoreductases with high affinity for phenolic polymers such as lignin. Based on high activity obtained from solid-state fermentation using a lignocellulose source from oil palm as matrix, we describe the isolation and whole-genome sequencing of Dictyopanus pusillus, a wild basidiomycete fungus exhibiting ABTS oxidation as an indication of laccase activity. Functional characterization of a crude enzymatic extract identified laccase activity as the main enzymatic contributor to fungal extracts, an observation supported by the identification of 13 putative genes encoding for homologous laccases in the genome. To the best of our knowledge, this represents the first report of an enzymatic extract exhibiting laccase activity in the Dictyopanus genera, offering means to exploit this species and its enzymes for the delignification process of lignocellulosic by-products from oil palm.

Retinal vein occlusion and paroxysmal nocturnal hemoglobinuria.

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare disorder associated with increased risk for thrombosis and reduced life expectancy. Retinal vein occlusion (RVO) is a frequent cause of vision loss but its relationship with PNH has not been studied systematically. Patients followed up for RVO in our ophthalmology department were screened for the presence of a PNH clone in peripheral blood by means of flow cytometry. The presence of other well-documented risk factors for RVO was also analyzed. In a series of 110 patients (54 males, median age of 67) we found no evidence of PNH. Most patients (97/110) had cardiovascular risk factors and/or hyperhomocysteinemia (67/110). Inherited thrombophilias were rare (three confirmed cases). Therefore, PNH does not appear to play a role in the development of RVO. However, this finding does not necessarily apply to young patients and/or those with no conventional risk factors for RVO, due to the low number of patients in these subgroups in our population.

Chronic proton pump inhibition therapy in the diagnostic accuracy of serum pepsinogen I and gastrin concentrations to identify pernicious anaemia.

BACKGROUND: Chronic use of proton pump inhibitors (PPIs) leads to increases in gastrin and pepsinogen-I serum concentrations. AIM: To asses if chronic treatment with PPIs has an effect on serum gastrin and pepsinogen-I concentrations for the diagnosis of pernicious anaemia (PA). MATERIALS AND METHODS: Serum gastrin and pepsinogen-I were measured in 38 patients with PA and 74 without PA (controls); 17/38 PA patients and 36/74 controls were treated with PPIs. Receiver Operating Curves (ROC) were used to compare diagnostic accuracy of gastrin and pepsinogen-I for PA in patients under chronic treatment with PPIs and in untreated patients. RESULTS: PPI treatment increased pepsinogen-I in patients and in controls, while gastrin increased only in controls. In untreated patients, a pepsinogen-I <8.3ng/mL had 95.2% sensitivity and 100% specificity, whereas a gastrin >115pg/mL had 100% sensitivity and 92.11% specificity for PA diagnosis. In PPI-treated patients, a pepsinogen I<24.1ng/mL had a lower sensitivity (82.4%) but retained 100% specificity, however the best cut-off point for gastrin, 610pg/mL, had a very low sensitivity (58%). CONCLUSIONS: PPI chronic treatment decreased the diagnostic accuracy for the studied biomarkers, particularly of gastrin. In PPI-treated patients, serum pepsinogen-I concentrations >24.1ng/mL allowed rejecting a PA diagnosis with 100% specificity.

Immunophenotypic, cytogenetic, and mutational characterization of cell lines derived from myelodysplastic syndrome patients after progression to acute myeloid leukemia.

Leukemia cell lines have been widely used in the hematology field to unravel mechanistic insights and to test new therapeutic strategies. Myelodysplastic syndromes (MDS) comprise a heterogeneous group of diseases that are characterized by ineffective hematopoiesis and frequent progress to acute myeloid leukemia (AML). A few cell lines have been established from MDS patients after progression to AML but their characterization is incomplete. Here we provide a detailed description of the immunophenotypic profile of the MDS-derived cell lines SKK-1, SKM-1, F-36P; and MOLM-13. Specifically, we analyzed a comprehensive panel of markers that are currently applied in the diagnostic routine for myeloid disorders. To provide high-resolution genetic data comprising copy number alterations and losses of heterozygosity we performed whole genome single nucleotide polymorphism-based arrays and included the cell line OHN-GM that harbors the frequent chromosome arm 5q deletion. Furthermore, we assessed the mutational status of 83 disease-relevant genes. Our results provide a resource to the MDS and AML field that allows researchers to choose the best-matching cell line for their functional studies. © 2016 Wiley Periodicals, Inc.

CD34 expression and the outcome of nucleophosmin 1-mutated acute myeloid leukemia.

CD34 positivity has been considered as an adverse prognostic factor in acute myeloid leukemia (AML). Although nucleophosmin 1-mutated (NPM1m) AML is usually CD34 negative, this marker may be expressed at diagnosis or acquired at relapse in a variable number of cases. Our objective was to ascertain if CD34 expression has any influence on the general outcome of this form of acute leukemia. Analysis of clinical outcome (complete remissions, relapses, disease-free survival, and overall survival) was performed depending on the degree of expression of CD34 determined by flow cytometry, in 67 adult patients with NPM1m AML. CD34 expression did not have any influence on the variables analyzed whatever the percentage of blasts expressing this marker. In contrast to other forms of AML, CD34 expression is not an unfavorable prognostic factor in NPM1m AML, neither at diagnosis nor at relapse.

Disminución de la respuesta eritropoyética en pacientes con anorexia nerviosa y anemia / Blunted erythropoietic response in the anemia of anorexia nervosa

Fundamento y objetivo: La causa o causas de la anemia que acompaña a la anorexia nerviosa (AN) no ha sido establecida, pero no parece relacionarse con deficiencias nutricionales ni cambios medulares. El objetivo de este trabajo fue evaluar la producción de eritropoyetina (EPO) en respuesta a la anemia en un pequeño grupo de pacientes con AN y anemia. Pacientes y métodos: Los niveles de EPO en muestras de suero de 41 mujeres con AN (11 con anemia y 30 sin alteraciones en los parámetros de la serie eritroide) se compararon con la respuesta observada en un grupo de pacientes de peso normal con anemia. Resultados: Las concentraciones de EPO en pacientes con AN anémicas fueron mayores que en las no anémicas: 20,63 mU/ml (4,04 a 28,46) frente a 8,7 mU/ml (3,9 a 20,93), p = 0,0088, pero el aumento de EPO fue menor de lo esperado (27,85 mU/ml [17,7 a 118,9]), p = 0,014. La correlación entre el IMC y la diferencia entre la EPO y la EPO esperada es inversa. Conclusiones: Una producción inadecuada de EPO puede explicar en parte la anemia en la AN. Son necesarios más estudios para investigar la causa de esta respuesta (AU)

Background and objective: The cause of the anemia in anorexia nervosa (AN) has not been fully ascertained. Ferritin, folate and cobalamin values are usually within normal ranges. Anemia does not have a relationship with bone marrow changes and erythropoietin (EPO) levels have not been investigated. The objective of this study was to evaluate the EPO response in a small group of AN patients. Patients and methods: EPO levels were measured in serum samples of 41 female AN patients (11 with anemia, and 30 with normal blood cell count). The adequacy of EPO response was assessed by comparing the increase observed in a group of normal weight patients with anemia. Results: EPO concentrations in anemic AN patients were higher than in non-anemic: 20.63 mU/mL (4.04-28.46) vs 8.7 mU/mL (3.9-20.93), P = .0088, but the increase in EPO was lower than expected (27.85 mU/mL [17.7-118.9]), P = .014. BMI and the difference between actual and expected EPO were inversely correlated. Conclusions: Inadequate EPO response may partly explain anemia in AN, but further studies are necessary (AU)

[Blunted erythropoietic response in the anemia of anorexia nervosa]. / Disminución de la respuesta eritropoyética en pacientes con anorexia nerviosa y anemia.

BACKGROUND AND OBJECTIVE: The cause of the anemia in anorexia nervosa (AN) has not been fully ascertained. Ferritin, folate and cobalamin values are usually within normal ranges. Anemia does not have a relationship with bone marrow changes and erythropoietin (EPO) levels have not been investigated. The objective of this study was to evaluate the EPO response in a small group of AN patients. PATIENTS AND METHODS: EPO levels were measured in serum samples of 41 female AN patients (11 with anemia, and 30 with normal blood cell count). The adequacy of EPO response was assessed by comparing the increase observed in a group of normal weight patients with anemia. RESULTS: EPO concentrations in anemic AN patients were higher than in non-anemic: 20.63mU/mL (4.04-28.46) vs 8.7mU/mL (3.9-20.93), P=.0088, but the increase in EPO was lower than expected (27.85mU/mL [17.7-118.9]), P=.014. BMI and the difference between actual and expected EPO were inversely correlated. CONCLUSIONS: Inadequate EPO response may partly explain anemia in AN, but further studies are necessary.

Blastocystis Isolates from Patients with Irritable Bowel Syndrome and from Asymptomatic Carriers Exhibit Similar Parasitological Loads, but Significantly Different Generation Times and Genetic Variability across Multiple Subtypes.

Blastocystis spp is a common intestinal parasite of humans and animals that has been associated to the etiology of irritable bowel syndrome (IBS); however, some studies have not found this association. Furthermore, many biological features of Blastocystis are little known. The objective of present study was to assess the generation times of Blastocystis cultures, from IBS patients and from asymptomatic carriers. A total of 100 isolates were obtained from 50 IBS patients and from 50 asymptomatic carriers. Up to 50 mg of feces from each participant were cultured in Barret's and in Pavlova's media during 48 h. Initial and final parasitological load were measured by microscopy and by quantitative PCR. Amplicons were purified, sequenced and submitted to GenBank; sequences were analysed for genetic diversity and a Bayesian inference allowed identifying genetic subtypes (ST). Generation times for Blastocystis isolates in both media, based on microscopic measures and molecular assays, were calculated. The clinical symptoms of IBS patients and distribution of Blastocystis ST 1, 2 and 3 in both groups was comparable to previous reports. Interestingly, the group of cases showed scarce mean nucleotide diversity (π) as compared to the control group (0.011±0.016 and 0.118±0.177, respectively), whilst high gene flow and small genetic differentiation indexes between different ST were found. Besides, Tajima's D test showed negative values for ST1-ST3. No statistical differences regarding parasitological load between cases and controls in both media, as searched by microscopy and by qPCR, were detected except that parasites grew faster in Barret's than in Pavlova's medium. Interestingly, slow growth of isolates recovered from cases in comparison to those of controls was observed (p<0.05). We propose that generation times of Blastocystis might be easily affected by intestinal environmental changes due to IBS probably because virulent strains with slow growth may be selected, reducing their genetic variability.

Correlation of CD11b and CD56 expression in adult acute myeloid leukemia with cytogenetic risk groups and prognosis.

Among other phenotypic markers, CD11b expression has been considered as an unfavorable prognostic factor, both in terms of overall survival (OS), disease-free survival (DFS), and attainment and duration of complete remissions (CRs) in adult patients with acute myeloid leukemia (AML). Recently, some groups have restricted its prognostic impact to poor prognostic karyotypic risk groups. The aim of this study was to retrospectively analyze the prevalence of CD11b and of CD56 expression in blast cells of 158 AML patients [excluding those with t(15;17)] stratified according to their cytogenetic risk and to correlate these phenotypic characteristics with OS, DFS, and CR. CD11b was more frequently expressed in intermediate and unfavorable cytogenetic prognostic groups (38.9 and 35.5 %, respectively) than in the favorable group (9.5 %). No differences were observed in CD56 expression according to the cytogenetic risk groups. When OS, DFS, and CR were analyzed according to these two markers, no statistical differences were recorded in any cytogenetic risk group. In conclusion, although CD11b was more frequently expressed in blast cells of patients with intermediate and unfavorable cytogenetic risk groups, this feature did not translate into different clinical outcome. Similarly, CD56 positivity did not have any influence on the prognosis of these patients.

Tributo a la vida y la obra del Dr Orlando Velázquez Martínez / Tribute to Dr Orlando Velazquez Martinez's life and work

En 2014 se cumplen 70 años del nacimiento y 10 años de la desaparición física del Dr Orlando Velázquez Martínez. Con el objetivo de rendirle un merecido homenaje, se realizó este trabajo. Se abordaron aspectos de su vida y obra, resaltándose sus valores y ejemplo, su trayectoria y las aristas más destacadas de su formación y desempeño como médico y profesor universitario, así como su amor por la Pediatría, la Medicina General Integral y su apego a los principios de la Revolución(AU)

There is going to be 70 years from birth and 10 years from decease of Dr Orlando Velazquez Martínez in 2014. With the objective of rendering the earned tribute, we developed this work. Several aspects of his life and work are dealt with, highlighting his values and example, his life course and the most important facets of his formation and work as a physician and university teacher, and also his love for Pediatrics, General Integral Medicine and his fondness of the principles of the Revolution(AU)