Evolution of Status of Trace Elements and Metallothioneins in Patients with COVID-19: Relationship with Clinical, Biochemical, and Inflammatory Parameters

The inflammatory reaction and pathogenesis of COVID-19 may be modulated by circulating trace elements (Iron (Fe), Zinc (Zn), Copper (Cu), Manganese (Mn)) and Metallothioneins (MTs). Thus, the present study aimed to investigate their relationship with clinical, biochemical, and inflammatory parameters in patients with COVID-19 at the early Intensive Care Unit (ICU) phase. Critically ill patients (n = 86) were monitored from the first day of ICU admission until the third day of stay. Serum samples were used to assess mineral levels via Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and MT levels via differential pulse voltammetry. Levels of Cu and MTs were significantly decreased after 3 days (p < 0.05), increasing the prevalence of Cu-deficient values from 50% to 65.3% (p = 0.015). Fe and Zn were shown to have a predictive value for mortality and severity. The present study suggests trace element deficiency may be a risk factor during early ICU treatment of COVID-19, as it is related to different biochemical and clinical parameters, indicating a possible beneficial effect of restoring proper levels of these micronutrients.European Regional Development Fund (ERDF) “a way of making Europe”Consejería de Transformación Económica, Industria, Conocimiento y Universidades de la Junta de Andalucía (REF. A-CTS-708-UGR20)Project FIS PI10/1993 from the Spanish Carlos III Health Institute(REF. FPU18/03702)(REF. FPU18/03655)Spanish Ministry of Educatio

Effect of zinc supplementation on circulating concentrations of homocysteine, vitamin B12, and folate in a postmenopausal population

Introduction: The decrease in estrogen levels associated with menopause increases the risk of deficiencies of key micronutrients such as zinc and of disturbances in methylation cycle-related markers. The present study assesses the effect of 8-week Zn supplementation upon circulating concentrations of Hcy, B12, and Fol levels in a population of postmenopausal women. Methods: Fifty-one postmenopausal women aged between 44 and 76 years took part in the study. Two randomized groups (placebo and zinc [50 mg/day]) were treated during 8 weeks. Nutrient intake was assessed based on the 72-hour recall method. Zinc was analyzed by flame atomic absorption spectrophotometry. Clinicalnutritional parameters were determined by enzyme immunoassay techniques. Results: Folate levels increased significantly (p < 0.05) in the zinc group on comparing the baseline versus followup values. Homocysteine decreased in the inter-group analysis (p < 0.05) after the intervention. Furthermore, higher folate (r = - 0.632; p = 0.005) and vitamin B12 (r = - 0.512; p = 0.030) levels were correlated to low homocysteine levels in the zinc group after the intervention, although the zinc intervention had the same effect on B12 levels in both groups. Conclusion: Zinc supplementation enhanced circulating folate and homocysteine by improving the folate values in the zinc-supplemented group and decreasing homocysteine levels inter-groups. Further studies involving larger samples and optimizing the doses and intervention period are needed to reinforce our main findings.Thanks are due to all the postmenopausal women who participated in the present study. We also thank the English revision and edition provided by a scientific translator. Funding for open access charge: Universidad de Huelva, Spain / CBUA

Evolution of Vitamin D Status and Vitamin D Receptor Gene Expression Among Professional Handball Athletes During a Competitive Period. Relationship with Body Composition, Calcium, Magnesium and Phosphorous

Introduction: A generalized risk of vitamin D deficiency exists worldwide affecting also professional and elite athletes. This study assesses the evolution of vitamin D status and vitamin D receptor (VDR) gene expression and their relationship with body composition, calcium (Ca), magnesium (Mg) and phosphorous (P) among professional handball athletes during a competitive period. Methods: A total of 26 male subjects were recruited: 13 professional handball athletes and 13 non-athlete controls. An observational follow-up study was conducted in 2 time points over a 16-week period. Nutritional intake, body composition, and routinary biochemical parameters were measured via 24-hours recall, bioimpedance and enzyme immunoassay, respectively. Ca and Mg were measured by flame atomic absorption spectrophotometry and P was determined with the colorimetric method of Fiske-Subbarow. 25-hydroxyvitamin-D (25(OH)D) levels and its forms (i.e., 25(OH)D3 and 25(OH)D2) were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS), whereas VDR gene expression was measured by quantitative real time-polymerase chain reaction (qRT-PCR). Results: A total of 54% of the athletes showed deficient vitamin D status. Moreover, a prevalence of insufficient vitamin D status in handball players affected 46% at baseline, reaching 61% after 16 weeks. Vitamin D showed no evolution during the competitive period and no differences between groups were observed (all p ≥ 0.05). Handball players increased the VDR expression, enhanced body composition, Ca and Mg levels at 16-weeks follow-up (all p < 0.05). VDR gene expression was positively related with body mass and body mass index at follow-up in athletes (all p ≤ 0.038; r ≥ 0.579) and with Ca at baseline in controls (p = 0.026; r = 0.648). Finally, 25(OH)D2 form was directly associated with P in athletes at 16 weeks of study (p = 0.034; r = 0.588). Conclusion: Players of indoor team sports such as handball would be a population at risk of vitamin D deficiency. The 16-weeks competition improved VDR gene expression, body composition, Ca and Mg levels. The associations observed between VDR gene expression and the variables of the study evidenced the importance of this receptor as a marker involved in health status in handball athletes despite vitamin D − although in a deficient status −, Ca, Mg and P showed no remarkable changes during the competition period.Spanish Ministry of Education (grant number AP2009-3701)FIS Project PI10/1993 from the Carlos III Health Institute (Spain)FPU fellowship from the Spanish Ministry of Education and Professional Formation with grant reference FPU18/03655 and FPU18/03702Funding for open access charge: Universidad de Granada/CBU

El ácido fólico y la vitamina B12 como biomarcadores de morbilidad y mortalidad en pacientes con shock séptico

Funding: financial support for the study was provided by Project FIS PI10/1993 from the Spanish Carlos III Health Institute and by the European Regional Development Fund. Lourdes Herrera-Quintana and Héctor Vázquez-Lorente are under a FPU fellowship from the Spanish Ministry of Education.Introduction and objective: a study was made of the folic acid (Fol) and vitamin B12 (B12) serum concentrations in critical patients with septic shock upon admission and after three days of stay in the Intensive Care Unit (ICU), with an analysis of their association to inflammatory parameters and patient morbidity-mortality. Methods: a prospective analytical study was made of 30 critically ill patients with septic shock. Demographic data, comorbidities, clinical information and severity scores were recorded. Data collected included serum Fol and B12 levels using the DxI® Autoanalyzer (Beckman Coulter) based on a competitive electrochemoluminescence immunoassay. Results: mean serum Fol was within the reference range stipulated by the laboratory on the first day. Nevertheless, a total of 21.4 % of the patients had high Fol levels, with 14.2 % being Fol deficient. An association was observed between Fol (p < 0.012) status and 28-day mortality, and the number of days of mechanical ventilation, fraction of inspired oxygen (FiO2 ) and fibrinogen increased in patients with higher Fol levels (p < 0.05). In addition, 85.7 % of cases had B12 levels above the reference values, with a correlation being observed between B12 and Fol. Conclusions: this study proposes Fol as a novel morbidity-mortality biomarker in critical septic patients, and reinforces the usefulness of B12 as a morbidity biomarker. It is thus suggested that the measurement of Fol upon admission and over the first 72 hours of hospital stay could provide prognostic information about the clinical course and outcome of septic shock patients.Introducción y objetivo: se realizó un estudio de las concentraciones séricas de ácido fólico (Fol) y vitamina B12 (B12) en pacientes críticos con shock séptico al ingreso y después de tres días de estancia en la Unidad de Cuidados Intensivos (UCI), con un análisis de su asociación con los parámetros inflamatorios y la morbimortalidad de los pacientes. Método: se realizó un estudio analítico prospectivo de 30 pacientes críticos con shock séptico. Se registraron datos demográficos, comorbilidades, información clínica y puntuaciones de gravedad. Los datos recopilados incluyeron los niveles séricos de Fol y B12 utilizando el autoanalizador DxI® (Beckman Coulter) basado en un inmunoensayo de electroquimioluminiscencia competitivo. Resultados: la media de Fol sérico estuvo dentro del rango de referencia estipulado por el laboratorio el primer día. Sin embargo, el 21,4 % de los pacientes presentaban niveles altos de Fol y el 14,2 % presentaban deficiencia de Fol. Se observó una asociación entre el estado de Fol (p < 0,012) con la mortalidad a los 28 días, con el número de días de ventilación mecánica, con la fracción de oxígeno inspirado (FiO2 ) y con el fibrinógeno, que aumentaron en los pacientes con niveles de Fol más altos (p < 0,05). Además, el 85,7 % de los casos tenían niveles de B12 por encima de los valores de referencia, observándose una correlación entre B12 y Fol. Conclusiones: este estudio propone al Fol como nuevo biomarcador de morbimortalidad en los pacientes críticos con sepsis y refuerza la utilidad de la B12 como biomarcador de morbilidad. Por tanto, se sugiere que la medición de Fol al ingreso y durante las primeras 72 horas de estancia hospitalaria podría proporcionar información pronóstica sobre el curso clínico y el resultado de los pacientes con shock séptico.Spanish Carlos III Health InstituteMinisterio de Educación, Cultura y DeporteEuropean Regional Development Fun

Relationship between Body Composition and Biochemical Parameters with Antioxidant Status in a Healthy Cohort of Postmenopausal Women

An adequate prooxidant-antioxidant balance-which may be influenced by body composition and biochemical status-is essential to maintain human health, especially in circumstances under which the antioxidant defense decreases, such as menopause. The present study aimed to examine the relationship between body composition and biochemical parameters with antioxidant status in a healthy cohort of postmenopausal women. This cross-sectional study was carried out in a cohort of 78 postmenopausal women aged 44-76 years. The body composition profile was assessed through bioelectrical impedance. The determination of the total antioxidant capacity and superoxide dismutase activity was conducted by the colorimetric method, and glutathione peroxidase activity was determined by the enzymatic immunological method. The vitamin D levels were measured by ultra-performance liquid chromatography-tandem mass spectrometry. The mineral status was assessed through flame atomic absorption spectrophotometry. The rest of the biochemical parameters were assessed through an immunoassay. The total antioxidant capacity and antioxidant gap were negatively influenced by body composition (all p <= 0.049) and positively related to protein metabolism parameters (all p <= 0.048), whereas circulating levels of different micronutrients (all p <= 0.048) and enzymes (all p <= 0.047) appeared to play an important role in the glutathione peroxidase and superoxide dismutase activities. In conclusion, the menopause-related antioxidant status changes may be influenced by key body composition and biochemical profiles. To confirm this statement, further trials aiming to evaluate the body composition and biochemical intervention-induced changes upon antioxidant defense are needed.Spanish Ministry of Education and Professional Formation FPU18/03655 FPU18/0370

Vitamin D Status in Critically Ill Patients with SIRS and Its Relationship with Circulating Zn and Related Parameters during ICU Stay

Critically ill patients are exposed to different stressors which may generate Systemic Inflammatory Response Syndrome (SIRS). This situation hinders the assessment of micronutrients status, such as vitamin D or Zinc (Zn), potentially affecting patients’ treatment and recovery. The aim of the present study was to assess the evolution of circulating 25–Hydroxyvitamin D (25–OH–D) levels after seven days of Intensive Care Unit (ICU) stay and the influence on changes in plasma and erythrocyte Zn levels, as well as other parameters related to phosphorus–calcium metabolism. A prospective analytical study was conducted on 65 critically ill patients (42% women) aged 31–77 years with SIRS. Total 25–OH–D levels were measured in plasma samples by liquid chromatographytandem mass spectrometry, and Zn content was analyzed by flame atomic absorption spectrometry. Both 25–OH–D and 25–OH–D3 levels were directly associated with erythrocyte Zn concentration at follow-up (p = 0.046 and p = 0.011, respectively). A relationship between erythrocyte and plasma Zn was also found at this follow-up point. No such clear associations were found when considering 25– OH–D2. Different disturbances in levels of phosphorus–calcium metabolism parameters were found, suggesting a relationship between the changes of 25–OH–D3 levels and parathormone (p = 0.019) and phosphorus (p = 0.005). The findings of the present study suggest an interaction between vitamin D and Zn, in which the correct status of these micronutrients could be a potentially modifiable factor and a beneficial approach in the recovery of critically ill patients.FIS projects of the Carlos III Health Institute PI10/1993Spanish Government A-CTS-708-UGR20 FPU18/03702 FPU18/0365

Zinc transporters expression profile in professional handball players supplemented with zinc

Introduction: Zinc (Zn) deficiency has been described not only on general human health but also within the sports context -as negatively affecting performance-. Thus, Zn status assessment is of great interest for athletes, especially in order to correct deficiency states of this mineral. Objective: The overall objective of this work was to assess Zn status in professional handball players during the competitive period (through plasma levels, dietary intake and gene expression of the Zn transporters), as well as to determine the effect of Zn supplementation. Methods: A total of twenty-two participants were recruited, -twelve belonged to the Control Group (CG) and ten male handball players comprised the experimental group (ATH-G)-, being monitored over a 2-month period with 2 evaluation moments: baseline (i.e., initial conditions) and follow-up (i.e., after 8 weeks of training and competition). Zn intake, plasma Zn levels, and gene expression of Zn transporters were obtained. Results: Plasma Zn levels were higher in ATH-G than in CG at the end of Zn intervention (p ≤ 0.010). Moreover, differences in the gene expression profile of Zn transporters were observed in ATH-G -with the down-regulation of several Zn transporters-, compared to the CG at baseline (p ≤ 0.05). Likewise, differences in the Zn transporters expression were observed in ATH-G at 8 weeks (all, p ≤ 0.001) -with ZnT2, ZnT5, ZIP3, ZIP5, ZIP11, ZIP13 and ZIP14 transporters being up-regulated-. Conclusion: Handball players seemed to have different nutritional needs for Zn, with differences in the gene expression of Zn transporters compared to controls. Zn intervention in our athletes may have influenced the expression of Zn transporters, indicating a potential increase in Zn transporters expression to mobilize Zn at the cellular level at 8 weeks of Zn intervention.We thank COST Action TD1304 e-Network for the Biology of Zinc (Zinc-Net) for favoring our collaboration with experts on zinc biomarkers. This article is based upon work from COST Action TD1304 supported by COST (European Cooperation in Science and Technology). Funding for open access charge: Universidad de Granada /CBUA.Departamento de Educación Física, Música y Artes Plástica

Erythrocyte Zn concentration and antioxidant response after supplementation with Zn in a postmenopausal population. A double-blind randomized trial

Background & aims: Menopausal hormonal changes increase the risk of deficiencies of minerals such as zinc (Zn), which could further worsen the decreased antioxidant defense of postmenopausal women. This study assesses the effect of 8 weeks of Zn supplementation upon the antioxidant status and clinical nutritional parameters of a postmenopausal population. Methods: Fifty-one postmenopausal women were divided into two groups: placebo (PbG) and zinc supplementation (ZnG). Mineral status was determined by Flame Atomic Absorption Spectrophotometry (FAAS). Total Antioxidant Capacity (TAC) and Superoxide Dismutase (SOD) were analyzed by kinetic colorimetric methods. Glutathione Peroxidase (GPx) was assessed by an enzymatic immunological method. Results: Poor Zn status was initially observed in erythrocyte samples. Total antioxidant capacity showed a significant correlation (r = 0.730; p < 0.05) to erythrocyte Zn after the intervention (ZnG: r = 0.96; p < 0.001). Moreover, erythrocyte Zn concentration in ZnG was positively correlated to GPx activity after the intervention (r = 0.61; p < 0.01). Conclusions: The postmenopausal women initially presented Zn deficiency, and the status of this mineral improved after the intervention. Zinc supplementation may be an effective approach for correcting the observed deficiencies, enhancing antioxidant defense in this risk population.We wish to thank all the women volunteers taking part in this study. We also acknowledge the expertise translator who provided English editing. Funding for open access charge: Universidad de Huelva/CBUA

Association between Body Fatness and Vitamin D3 Status in a Postmenopausal Population

We especially thank all the postmenopausal women who participated in our study and Bienza translators who provided English editing.Vitamin D is a micronutrient that plays a key role in phosphocalcic metabolism. The postmenopausal population presents a risk of deficiency in this vitamin due to hormonal alterations which, in the case of obesity, would be exacerbated. The objective was to assess the status of vitamin D in a postmenopausal population and determine the relationship of 25-hydroxivitamin D [25(OH)D] and its metabolites with anthropometric parameters. The study included 78 healthy postmenopausal women aged from 44 to 76. The nutrient intake assessment was carried out using the 24 h reminder (R24h). 25(OH)D was analyzed using ultra-high-performance liquid chromatography (UHPLC). A total of 80% and 68% of the women studied did not reach sufficient values of 25(OH)D and 25-hydroxivitamin D3 [25(OH)D3], respectively, which was inversely correlated with Body Mass Index (BMI) (r = −0.25, p = 0.04), hip perimeter (r = −0.26 and r = −0.24, all p < 0.05), arm circumference (r = −0.29, p = 0.01) and fat mass (r = −0.28 and r = −0.26, all p < 0.05). 25(OH)D3 is the metabolite that contributed most to this association. In conclusion, 25(OH)D3 levels are related to anthropometric parameters in the postmenopausal women in this study, confirming insufficient status in the majority of the population. Approach strategies are necessary to correct and avoid this risk in order to ensure future quality of life.This research was funded by FIS Carlos III (REF. PI10/1993)

Pyroglutamic acidosis by glutathione regeneration blockage in critical patients with septic shock

Aim: The aim of this study was to evaluate oxidative stress from glutathione depletion in critically ill patients with a septic shock through the abnormal presence of pyroglutamic acid (PyroGlu) in the urine (indirectly) and through its serum level (directly). Methods: This was a prospective analytical study of 28 critically ill patients with a septic shock who were monitored from admission (initial) to 3 days of stay (final) in the intensive care unit (ICU). Data collected included PyroGlu and glutamic acid (Glu) using liquid chromatography/mass spectrometry, and glutathione peroxidase (GPX) activity with a colorimetric assay. The differences in Glu, PyroGlu, and GPX activity between the septic shock group and healthy control group serving as reference values were evaluated using the Mann–Whitney test. The correlations between Glu, PyroGlu, and GPX activity and clinical outcomes were determined using Spearman’s correlation coefficient. Results: In patients with septic shock, serum and urine PyroGlu levels were higher, erythrocyte GPX activity/gr Hb was lower, and urine Glu levels were lower compared to healthy control reference values, for both initial and final values. Initial serum Glu levels were also lower. Serum PyroGlu levels had a correlation with both initial and final serum Glu levels; levels also correlated in the urine. Initial serum Glu correlated with the days of mechanical ventilation (P = 0.016) and the days of ICU stay (P = 0.05). Urine Glu/mg creatinine correlated with APACHE II (P = 0. 030). This positive correlation observed for serum Glu was not observed for PyroGlu. Conclusions: The current study found that septic patients have higher levels of PyroGlu, lower levels of Glu, and lower erythrocyte GPX activity, suggesting that these biomarkers could be used as an indicator of glutathione depletion. In addition, Glu is related to severity parameters. This study can guide future studies on the importance of monitoring the levels of pyroglutamic acidosis in critical patients with septic shock in order to preserve the oxidative status and its evolution during the stay in the ICU.Financial support for the study was provided by Project FIS PI10/1993 from the Spanish Carlos III Health Institute and FEDER European Funds