CORRELATION BETWEEN PLASMATIC VITAMIN D LEVEL AND REFRACTIVE STATUS IN CHILDREN WITH DISABILITIES

Introduction. Literature confirms that refractive errors are the most common, easily corrected, eye morbidity in children with disabilities. Early intervention such as wearing eyeglasses can positively impact the lives of these children. The implication of vitamin D status is investigated as a possible proactive measure in eye conditions. Aim. The current study proposed to asses the refractive status as well as vitamin D plasmatic level in 161 children. Another aim was to investigate whether myopia corelates with a lower plasmatic vitamin D level. Methods. A retrospective case-control study was done on 161 children, divided into two groups: the study group (children with disabilities) and the contol group (children without disabilities). The age range of children included in the study was from 5 to 16 years old. Results. Refractive errors were found to be more frequent in the group of children with disabilities and of these, astigmatism was the most frequent refractive disorder identified. Also, the plasmatic vitamin D level was found to be lower in those with myopia reguardless of disability status. Conclusions. Children with disabilities are diagnosed with refractive errors twice more frequenty than their healthy peers. Parents, medical staff and teachers should be aware of this risk factor and be more attentive because the presence of uncorrected refractive disorders may not be visible in most children, especially those with special needs.

Vitamin D and Primary Ciliary Dyskinesia: A Topic to Be Further Explored

Primary ciliary dyskinesia (PCD) is a genetic disease characterized by abnormalities in ciliary structure/function. The diagnosis of PCD relies on a combination of clinical evaluation and ultrastructural (electron microscopic) analysis of the ciliary architecture. This diagnosis may be challenging due to clinical and genetic heterogeneity and artifacts during the ciliary ultrastructure preparation and assessment. Recently, vitamin D supplementation has been proposed for several groups probably suffering from D-hypovitaminosis. Some patients with inflammatory bowel disease may have significant malabsorption, and vitamin D supplementation in these patients is recommended. Two recent reports suggest that a low plasmatic level of this vitamin is present in the PCD population. The utility of vitamin D supplementation may be essential in this group of individuals, and further investigations are warranted. Still, in examining the literature papers, it seems relevant that the authors concentrate solely on lung function in both studies. Future studies should probably target the intestinal function in patients with PCD independently from the vitamin D supplementation to fully evaluate its role.

Soluble BTN2A1 Is a Potential Prognosis Biomarker in Pre-Treated Advanced Renal Cell Carcinoma

The development of immune checkpoint inhibitors (ICI) has dramatically changed the landscape of therapies for metastatic renal cell carcinoma. However, many patients do not benefit from such therapy and prognostic or predictive validated biomarker validated for ICI are still needed to better select and treat patient. Plasmatic soluble immune checkpoints have been described as potential immune biomarkers in hematological malignancies and solids tumors, then, we would like to explore the prognostic value of different soluble immune checkpoints in patients with mRCC treated with nivolumab after TKI. We prospectively collected plasma samples before nivolumab infusion from 38 patients previously treated for mRCC with TKI at Paoli-Calmettes Institute, from the NIVOREN GETUG-AFU 26 study (NCT03013335). Enzyme-linked immunosorbent assays (ELISA) were performed for soluble forms of PD-1, PD-L1, global BTN3, BTLA, BTN3A1 and BTN2A1. Among the different soluble checkpoints analyzed, only high baseline plasmatic level of BTN2A1 was significantly associated with shorter PFS: median PFS was 3.95 months for sBTN2A1high vs 14.30 months for sBTN2A1low (sBTN2A1 cut-off: 6.7ng/mL; HR = 2.26, 95%CI [0.68 – 4.60], p = 0.0307). There was no statistical difference in OS between sBTN2A1high and sBTN2A1low. Our results suggest that the baseline level of plasmatic BTN2A1 could be an independent prognosis factor of PFS after nivolumab for pre-treated patient with mRCC. However, these results need to be validated in a larger prospective cohort and the biological role of BTN subfamily and γδ T cell immunity in mRCC must be elucidated.

Assessment of Factors Related to Diminished Appetite in Hemodialysis Patients with a New Adapted and Validated Questionnaire

Appetite loss is a common phenomenon in end-stage renal disease (ESRD) patients undergoing maintenance hemodialysis (HD). We aimed to (i) adapt and validate a Spanish language version of the Council on Nutrition Appetite Questionnaire (CNAQ) and (ii) to identify psychological and biological factors associated with diminished appetite. We recruited 242 patients undergoing HD from four hemodialysis centers to validate the Spanish-translated version of the CNAQ. In another set of 182 patients from three HD centers, the Appetite and Diet Assessment Tool (ADAT) was used as the gold standard to identify a cut-off value for diminished appetite in our adapted questionnaire. The Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Distorted Thoughts Scale (DTS), Dialysis Malnutrition Score (DMS), anthropometric, values and laboratory values were also measured. Seven items were preserved in the adapted appetite questionnaire, with two factors associated with flavor and gastric fullness (Cronbach’s alpha = 0.758). Diminished appetite was identified with a cut-off value ≤25 points (sensitivity 73%, specificity 77%). Patients with diminished appetite had a higher proportion of females and DMS punctuation, lower plasmatic level of creatinine, blood urea nitrogen, and phosphorus. Appetite score correlated with BDI score, BAI score and DTS. Conclusions: This simple but robust appetite score adequately discriminates against patients with diminished appetite. Screening and treatment of psychological conditions may be useful to increase appetite and the nutritional status of these patients.

Effect of Postbiotic Based on Lactic Acid Bacteria on Semen Quality and Health of Male Rabbits

The aim of this study was to evaluate the effect of lactic acid bacteria-based postbiotic supplementation on semen characteristics and hematological and biochemical profiles in rabbits. A total of 28 males were randomly allocated into two groups. Males received a Control diet and Enriched diet supplemented with postbiotic for 15 weeks (4 weeks of adaptation period and 11 weeks of experimental period). Body weight, feed intake and semen characteristics were recorded weekly. Hematological profile was recorded at the beginning and end of the experiment and biochemical profile at 0, 5, 10 and 15 weeks. Bayesian methodology was used for the statistical analysis. Feed intake was higher in Control diet (125.2 g) than in the Enriched diet (118.6 g, p = 1.00). The percentages of abnormal spermatozoa were higher in Control diet than in Enriched diet (30% and 22%; p = 0.93) and the acrosome integrity percentage was lower (97% and 96%; p = 0.87). The hematological profile was within the range for healthy rabbits. The plasmatic level of alanine aminotransferase was higher in Control diet than Enriched diet at 5 and 10 weeks (p = 0.93 and p = 0.94, respectively) and alkaline phosphatase was similar in Control diet throughout the experiment, but decreased in Enriched diet (p = 0.97). No difference was found in kidney parameters (uric nitrogen and creatinine). Enriched diet showed higher total protein and globulin than Control diet (p = 0.99). Phosphorus was lower (p = 0.92) in Control diet than in Enriched diet. In conclusion, the addition of the postbiotic based on lactic acid bacteria seems to improve the quality of the semen and the liver profile in rabbits.

Increased sCD163 and sCD14 Plasmatic Levels and Depletion of Peripheral Blood Pro-Inflammatory Monocytes, Myeloid and Plasmacytoid Dendritic Cells in Patients With Severe COVID-19 Pneumonia

BackgroundEmerging evidence argues that monocytes, circulating innate immune cells, are principal players in COVID-19 pneumonia. The study aimed to investigate the role of soluble (s)CD163 and sCD14 plasmatic levels in predicting disease severity and characterize peripheral blood monocytes and dendritic cells (DCs), in patients with COVID-19 pneumonia (COVID-19 subjects).MethodsOn admission, in COVID-19 subjects sCD163 and sCD14 plasmatic levels, and peripheral blood monocyte and DC subsets were compared to healthy donors (HDs). According to clinical outcome, COVID-19 subjects were divided into ARDS and non-ARDS groups.ResultsCompared to HDs, COVID-19 subjects showed higher sCD163 (p<0.0001) and sCD14 (p<0.0001) plasmatic levels. We observed higher sCD163 plasmatic levels in the ARDS group compared to the non-ARDS one (p=0.002). The cut-off for sCD163 plasmatic level greater than 2032 ng/ml was predictive of disease severity (AUC: 0.6786, p=0.0022; sensitivity 56.7% [CI: 44.1–68.4] specificity 73.8% [CI: 58.9–84.7]). Positive correlation between plasmatic levels of sCD163, LDH and IL-6 and between plasmatic levels of sCD14, D-dimer and ferritin were found. Compared to HDs, COVID-19 subjects showed lower percentages of non-classical (p=0.0012) and intermediate monocytes (p=0.0447), slanDCs (p<0.0001), myeloid DCs (mDCs, p<0.0001), and plasmacytoid DCs (pDCs, p=0.0014). Compared to the non-ARDS group, the ARDS group showed lower percentages of non-classical monocytes (p=0.0006), mDCs (p=0.0346), and pDCs (p=0.0492).ConclusionsThe increase in sCD163 and sCD14 plasmatic levels, observed on hospital admission in COVID-19 subjects, especially in those who developed ARDS, and the correlations of these monocyte/macrophage activation markers with typical inflammatory markers of COVID-19 pneumonia, underline their potential use to assess the risk of progression of the disease. In an early stage of the disease, the assessment of sCD163 plasmatic levels could have clinical utility in predicting the severity of COVID-19 pneumonia.

Persistance of Inflammatory Phenotype of Neutrophils in Sickle Cell Disease Children Despite Chronic Exchange Transfusion Program: A Cause of the Progression of Cerebral Vasculopathy?

Sickle cell disease (SCD) is a severe hemoglobinopathy due to the production of abnormal hemoglobin S (HbS). Although red blood cell (RBC) dysfunction is the major contributor to disease, several studies highlighted the important role of polymorphonuclear neutrophils (PMNs), both during acute and chronic complications. One of the most severe complication of SCD is ischemic stroke due to large cerebral artery occlusion. In 1998, the Stroke Prevention (STOP) trial demonstrated that monthly blood transfusions could reduce the risk of stroke by 90% in SCD children with cerebral vasculopathy (CV). However, there is a wide heterogeneity in the course of CV in patients receiving chronic transfusions, since only about half of them improved their CV under transfusion program, while 25% are only stabilized and 29% continue to get worse despite a percentage of HbS permanently below 30%. The aim of our study is to investigate the impact of transfusion programs on neutrophils activation and ageing, in order to identify if inflammation could contribute to the persistence of SCD complications despite red cell transfusion. We performed a prospective study including 58 homozygous SCD children and 10 healthy donors. Of these, 12 had no specific treatment, 11 were on Hydroxyurea (HU) treatment, 21 were on an exchange transfusion program, and 14 were on both an exchange transfusion program and HU treatment for an average of 4.9 years due to persistent CV. Monthly exchange transfusion are carried out either by erythrapheresis or by manual exchanges, consisting of the continuous bleeding of whole blood compensated by simultaneous transfusion of packed red blood cells. Neutrophils were isolated from fresh blood samples before exchange transfusion session and labelled with 8 markers specific of adhesion, activation and ageing. We quantified by flow cytometry the expression of 3 integrins (CD18, CD11a, CD11b), 3 ageing markers (CD182, CD184, CD62L) and 2 adhesion molecules (CD162 and CD66a). We also measured the plasmatic level of elastase, which reflects the NETose activity of PMNs As previously reported, we observed a high leukocytosis and an activated profile of PMNs in the 12 non-transfused SCD patients compared to healthy controls (Figure 1), characterized by an overexpression of the integrin CD18/CD11b (p=0,03) and CD18/CD11a (p=0,02), a higher level of circulating aged PMNs CD184 high/CD62Llow (p=0,04), a higher expression of CD162 (p=0,01) and CD66a (0,01) as well as a higher plasmatic level of elastase (p=0.01). Interestingly, in the PMNs of the 21 patients receiving monthly exchange transfusion, we found an identical expression pattern of integrins, selectins, ageing markers and elastase level compared to those of the PMNs from non-transfused patients. Furthermore, we also observed a persistence of high neutrophilic leukocytosis. This activation pattern was the same for patients on manual exchange or erythrapheresis, even with a tendency towards a more inflammatory profile in patients on erythrapheresis (Figure 1). In the PMNs from the 11 patients receiving HU compared to untreated SCD patients, we found an expected decrease in high leukocytosis and membrane integrin expression CD18/CD11b and CD18/CD11a. The addition of HU therapy in 14 patients in exchange transfusion program allows to alleviate neutrophilic leukocytosis and membrane integrin expression. Our study shows for the first time that replacing sickle RBCs with healthy RBCs is not sufficient to reverse the pathological phenotype of PMNs in SCD. A persistence of the PMNs activation pattern is observed both despite erythrapheresis, where plasma and white blood cells go back to the patient, and in manual exchanges, where the patient is bled from a large volume of whole blood. Given the major role of inflammation in endothelial damage and vasculopathy in SCD, our data could explain the incomplete efficacy of transfusion exchange programs to treat CV. This raises the question to systematically combine anti-inflammatory and anti-white blood cell adhesion treatments such as Hydroxyurea or P-Selectin inhibitors for these patients. Disclosures No relevant conflicts of interest to declare.

Matrix metalloproteainase-3 polymorphism association with hypertrophic cardiomyopathy

Background Matrix metalloproteinase-3 (MMP-3) level disequilibrium in hypertrophic cardiomyopathy (HCM) may be due to a specific genetic variation. The MMP-3 gene promoter contains an insertion/deletion polymorphism characterized by an array of 5 or 6 adenosine residues (5A/6A) at –1612 positions. Purpose The aim was to analyze whether the MMP-3 5A/6A gene promoter polymorphism is related to its level in HCM patients. Methods/Results In this study, we recruited 33 HCM patients and 35 non-HCM. The ELISA sandwich assay measured MMP-3 plasmatic level. The MMP-3 –1612 5A/6A polymorphism was genotyped by RFLP-PCR. The studied population was consistentin Hardy-Weinberg equilibrium. There were 17% 5A/5A homozygotes, 65% 6A/6A homozygotes, and 17% 5A/6A heterozygotes. The HCM was related to the existence of the –1612 5A/6A polymorphism (p<0.05). Patients carrying the 5A allele had a higher MMP-3 level than those with the 6A allele (16.03±9.43 vs 8.68±5.89 ng/ml, respectively; p=0.01). Conclusion Our data shows that the –1612 5A/6A MMP-3 gene polymorphism is associated to the hypertrophic cardiomyopathy and do influence the MMP-3 plasmatic circulating level. Funding Acknowledgement Type of funding source: None

Chronic hypoxaemia and gender status modulate adiponectin plasmatic level and its multimer proportion in severe COPD patients: new endotypic presentation?

Background Disease progression in COPD patient is associated to lung function decline, leading to a higher risk of hypoxaemia and associated comorbidities, notably cardiovascular diseases (CVD). Adiponectin (Ad) is an adipokine with cardio-protective properties. In COPD patients, conflicting results were previously reported regarding Ad plasmatic (Adpl) level, probably because COPD is a heterogeneous disease with multifactorial influence. Among these factors, gender and hypoxaemia could interact in a variety of ways with Ad pathway. Therefore, we postulated that these components could influence Adpl level and its multimers in COPD patients and contribute to the appearance of a distinct endotype associated to an altered CVD risk. Methods One hundred COPD patients were recruited: 61 were men and 39 were women. Patients who were not severely hypoxemic were allocated to non-hypoxemic group which included 46 patients: 27 men and 19 women. Hypoxemic group included 54 patients: 34 men and 20 women. For all patients, Adpl level and proportion of its different forms were measured. Differences between groups were evaluated by Rank-Sum tests. The relationship between these measures and BMI, blood gas analysis (PaO2, PaCO2), or lung function (FEV1, FEV1/FVC, TLCO, TLC, RV) were evaluated by Pearson correlation analysis. Results Despite similar age, BMI and obstruction severity, women had a higher TLC and RV (median: TLC = 105%; RV = 166%) than men (median: TLC = 87%; RV = 132%). Adpl level was higher in women (median = 11,152 ng/ml) than in men (median = 10,239 ng/ml) and was negatively associated with hyperinflation (R = − 0,43) and hypercapnia (R = − 0,42). The proportion of the most active forms of Ad (HMW) was increased in hypoxemic women (median = 10%) compared with non-hypoxemic women (median = 8%) but was not modulated in men. Conclusion COPD pathophysiology seemed to be different in hypoxemic women and was associated to Ad modulations. Hyperinflation and air-trapping in association with hypercapnia and hypoxaemia, could contribute to a modulation of Adpl level and of its HMW forms. These results suggest the development of a distinct endotypic presentation, based on gender.

Vitamin C, a Key Factor in Post-extraction Healings A Preliminary Study

In clinical practice, after dental extraction, the main objective is the healing of the post-extraction wound, without complications and in the shortest time possible. According to specialized studies, the defense and regeneration mechanisms of the body are in direct correlation with the plasmatic level of some vitamins, mainly vitamin C, which is directly involved in the repair of damaged tissues. The purpose of the present study was to evaluate the healing of the post-extraction wound in the conditions of oral administration of vitamin C, according to a predetermined administration protocol, as compared to the healing process under conventional conditions. Five volunteers were included in the study, who presented two teeth with indication of extraction, one of the extractions being performed under the conditions of administration of vitamin C. The evolution of the healing of the post-extraction wounds was assessed comparatively. The examination was done with the help of photographic examination by directly measuring the vestibulo-oral dimensions of the wounds using the digital micrometer and by recording the blood microcirculation at the level of the alveolar wound using the laser Doppler flowmetry. The level of vitamin C in the tissues was also determined, using the lingual test for vitamin C, a simple and non-invasive method that uses the reagent 2.6 dichlorophenol-indophenol. The results of the preliminary study show that the evolution and healing of the post-extraction wound is accelerated under the conditions of vitamin C administration, with regard to the administration protocol used in the study and compared to the healing situation under classical conditions. It can be stated that the human body consumes higher amounts of ascorbic acid in the case of dental extractions therefore the administration of vitamin C is a necessity if a faster healing of their post-extraction wounds is desired.