Corrigendum: Newborn screening for X-linked adrenoleukodystrophy in Italy: diagnostic algorithm and disease monitoring.

[This corrects the article DOI: 10.3389/fneur.2022.1072256.].

Fifteen Years of Iodine Prophylaxis in Italy: Results of a Nationwide Surveillance (Period 2015-2019).

CONTEXT: In 2005, a nationwide program of iodine prophylaxis on a voluntary basis was implemented in Italy by law. However, recent data on iodine status are lacking. OBJECTIVE: The aim of this study was to evaluate efficiency, effectiveness, and possible adverse effects (increased occurrence of thyroid autoimmunity and hyperthyroidism) of the Italian iodine prophylaxis program. METHODS: From 2015 to 2019, a nationwide survey was performed. The use of iodized salt was evaluated in a sample of 164 593 adults and in 998 school canteens. A sample of 4233 schoolchildren (aged 11-13 years) was recruited to assess urinary iodine concentration, prevalence of goiter, and thyroid hypoechogenicity on ultrasound, with the latter being an indirect indicator of thyroid autoimmunity. Neonatal TSH values of 197 677 infants screened in regions representative of Northern, Central, and Southern Italy were analyzed to investigate the percentage of TSH values >5.0 mIU/L. Data on methimazole prescriptions were analyzed as indirect indicators of new cases of hyperthyroidism. RESULTS: The prevalence of the use of iodized salt was 71.5% in adult population and 78% in school canteens. A median urinary iodine concentration of 124 µg/L, a prevalence of goiter of 2.2%, and a prevalence of thyroid hypoechogenicity of 5.7% were observed in schoolchildren. The percentage of neonatal TSH values >5.0 mIU/L resulted still higher (5.1%) than the World Health Organization threshold of 3.0%, whereas the prescriptions of methimazole showed a reduction of 13.5%. CONCLUSION: Fifteen years of iodine prophylaxis have led to iodine sufficiency in Italy, although there still is concern about iodine nutritional status during pregnancy.

Safety and Preliminary Efficacy of Mesenchymal Stromal Cell (ORBCEL-M) Therapy in Diabetic Kidney Disease: A Randomized Clinical Trial (NEPHSTROM).

SIGNIFICANCE STATEMENT: Mesenchymal stromal cells (MSCs) may offer a novel therapy for diabetic kidney disease (DKD), although clinical translation of this approach has been limited. The authors present findings from the first, lowest dose cohort of 16 adults with type 2 diabetes and progressive DKD participating in a randomized, placebo-controlled, dose-escalation phase 1b/2a trial of next-generation bone marrow-derived, anti-CD362 antibody-selected allogeneic MSCs (ORBCEL-M). A single intravenous (iv) infusion of 80×10 6 cells was safe and well-tolerated, with one quickly resolved infusion reaction in the placebo group and no subsequent treatment-related serious adverse events (SAEs). Compared with placebo, the median annual rate of decline in eGFR was significantly lower with ORBCEL-M, although mGFR did not differ. The results support further investigation of ORBCEL-M in this patient population in an appropriately sized phase 2b study. BACKGROUND: Systemic therapy with mesenchymal stromal cells may target maladaptive processes involved in diabetic kidney disease progression. However, clinical translation of this approach has been limited. METHODS: The Novel Stromal Cell Therapy for Diabetic Kidney Disease (NEPHSTROM) study, a randomized, placebo-controlled phase 1b/2a trial, assesses safety, tolerability, and preliminary efficacy of next-generation bone marrow-derived, anti-CD362-selected, allogeneic mesenchymal stromal cells (ORBCEL-M) in adults with type 2 diabetes and progressive diabetic kidney disease. This first, lowest dose cohort of 16 participants at three European sites was randomized (3:1) to receive intravenous infusion of ORBCEL-M (80×10 6 cells, n =12) or placebo ( n =4) and was followed for 18 months. RESULTS: At baseline, all participants were negative for anti-HLA antibodies and the measured GFR (mGFR) and estimated GFR were comparable between groups. The intervention was safe and well-tolerated. One placebo-treated participant had a quickly resolved infusion reaction (bronchospasm), with no subsequent treatment-related serious adverse events. Two ORBCEL-M recipients died during follow-up of causes deemed unrelated to the trial intervention; one recipient developed low-level anti-HLA antibodies. The median annual rate of kidney function decline after ORBCEL-M therapy compared with placebo did not differ by mGFR, but was significantly lower by eGFR estimated by the Chronic Kidney Disease Epidemiology Collaboration and Modification of Diet in Renal Disease equations. Immunologic profiling provided evidence of preservation of circulating regulatory T cells, lower natural killer T cells, and stabilization of inflammatory monocyte subsets in those receiving the cell therapy compared with placebo. CONCLUSIONS: Findings indicate safety and tolerability of intravenous ORBCEL-M cell therapy in the trial's lowest dose cohort. The rate of decline in eGFR (but not mGFR) over 18 months was significantly lower among those receiving cell therapy compared with placebo. Further studies will be needed to determine the therapy's effect on CKD progression. CLINICAL TRIAL REGISTRATION NUMBER: ClinicalTrial.gov NCT02585622 .

Betting on Non-Invasive Brain Stimulation to Treat Gambling Disorder: A Systematic Review and Meta-Analysis.

Gambling disorder (GD) is a behavioral addiction that severely impacts individuals' functioning, leading to high socioeconomic costs. Non-invasive brain stimulation (NiBS) has received attention for treating psychiatric and neurological conditions in recent decades, but there is no recommendation for its use for GD. Therefore, this study aimed to systematically review and analyze the available literature to determine the effectiveness of NiBS in treating GD. Following the PRISMA guidelines, we screened four electronic databases up to July 2022 and selected relevant English-written original articles. We included ten papers in the systematic review and seven in the meta-analysis. As only two studies employed a sham-controlled design, the pre-post standardized mean change (SMCC) was computed as effect size only for real stimulation. The results showed a significant effect of NiBS in reducing craving scores (SMCC = -0.69; 95% CI = [-1.2, -0.2], p = 0.010). Moreover, considering the GD's frequent comorbidity with mood disorders, we ran an exploratory analysis of the effects of NiBS on depressive symptoms, which showed significant decreases in post-treatment scores (SMCC = -0.71; 95% CI = [-1.1, -0.3], p < 0.001). These results provide initial evidence for developing NiBS as a feasible therapy for GD symptoms but further comprehensive research is needed to validate these findings. The limitations of the available literature are critically discussed.

Newborn screening for X-linked adrenoleukodystrophy in Italy: Diagnostic algorithm and disease monitoring.

Introduction: X-linked adrenoleukodystrophy (X-ALD) is the most common inherited peroxisomal disorder caused by variants in the ABCD1 gene. The main phenotypes observed in men with X-ALD are primary adrenal insufficiency, adrenomyeloneuropathy, and cerebral ALD (cALD). Cerebral ALD consists of a demyelinating progressive cerebral white matter (WM) disease associated with rapid clinical decline and is fatal if left untreated. Hematopoietic stem cell transplantation is the standard treatment for cALD as it stabilizes WM degeneration when performed early in the disease. For this reason, early diagnosis is crucial, and several countries have already implemented their newborn screening programs (NBS) with the assessment of C26:0-lysophosphatidylcholine (C26:0-LPC) values as screening for X-ALD. Methods: In June 2021, an Italian group in Lombardy launched a pilot study for the implementation of X-ALD in the Italian NBS program. A three-tiered approach was adopted, and it involved quantifying the values of C26:0-LPC and other metabolites in dried blood spots with FIA-MS/MS first, followed by the more specific ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) technique and, finally, the genetic confirmation via focused NGS. Discussion: Genetically confirmed patients are set to undergo a follow-up protocol and are periodically evaluated to promptly start a specific treatment if and when the first signs of brain damage appear, as suggested by international guidelines. A specific disease monitoring protocol has been created based on literature data and personal direct experience. Conclusion: The primary aim of this study was to develop a model able to improve the early diagnosis and subsequent follow-up and timely treatment of X-ALD. Ethics: The study was approved by the local ethics committee. The research was conducted in the absence of any commercial or financial relationship that could be construed as a potential conflict of interest.

Eculizumab in patients with severe coronavirus disease 2019 (COVID-19) requiring continuous positive airway pressure ventilator support: Retrospective cohort study.

BACKGROUND: Complement activation contributes to lung dysfunction in coronavirus disease 2019 (COVID-19). We assessed whether C5 blockade with eculizumab could improve disease outcome. METHODS: In this single-centre, academic, unblinded study two 900 mg eculizumab doses were added-on standard therapy in ten COVID-19 patients admitted from February 2020 to April 2020 and receiving Continuous-Positive-Airway-Pressure (CPAP) ventilator support from ≤24 hours. We compared their outcomes with those of 65 contemporary similar controls. Primary outcome was respiratory rate at one week of ventilator support. Secondary outcomes included the combined endpoint of mortality and discharge with chronic complications. RESULTS: Baseline characteristics of eculizumab-treated patients and controls were similar. At baseline, sC5b-9 levels, ex vivo C5b-9 and thrombi deposition were increased. Ex vivo tests normalised in eculizumab-treated patients, but not in controls. In eculizumab-treated patients respiratory rate decreased from 26.8±7.3 breaths/min at baseline to 20.3±3.8 and 18.0±4.8 breaths/min at one and two weeks, respectively (p<0.05 for both), but did not change in controls. Between-group changes differed significantly at both time-points (p<0.01). Changes in respiratory rate correlated with concomitant changes in ex vivo C5b-9 deposits at one (rs = 0.706, p = 0.010) and two (rs = 0.751, p = 0.032) weeks. Over a median (IQR) period of 47.0 (14.0-121.0) days, four eculizumab-treated patients died or had chronic complications versus 52 controls [HRCrude (95% CI): 0.26 (0.09-0.72), p = 0.010]. Between-group difference was significant even after adjustment for age, sex and baseline serum creatinine [HRAdjusted (95% CI): 0.30 (0.10-0.84), p = 0.023]. Six patients and 13 controls were discharged without complications [HRCrude (95% CI): 2.88 (1.08-7.70), p = 0.035]. Eculizumab was tolerated well. The main study limitations were the relatively small sample size and the non-randomised design. CONCLUSIONS: In patients with severe COVID-19, eculizumab safely improved respiratory dysfunction and decreased the combined endpoint of mortality and discharge with chronic complications. Findings need confirmation in randomised controlled trials.

Alpha defensin, leukocyte esterase, C-reactive protein, and leukocyte count in synovial fluid for pre-operative diagnosis of periprosthetic infection.

Synovial fluid analysis for diagnosis of prosthetic joint infections has gained increasing interest in the recent past when markers more specific for these infections than the serum ones have been identified. Despite the important steps forward, identification of a gold standard has not yet been identified. In this study, usefulness of alpha defensin, leukocyte esterase, C-reactive protein (CRP), and white blood cells (WBCs) in synovial fluids alone and in combination for diagnosis of prosthetic joint infection was evaluated. Synovial fluids from 32 infected and 34 not infected patients were analyzed. Sensitivity, specificity, positive and negative predictive values, diagnostic accuracy, and receiver-operating characteristic (ROC) curves were calculated for each parameter. Moreover, combination of coupled variables was also evaluated by logistic regression analysis. Sensitivity of alpha defensin, CRP, leukocyte count, and leukocyte esterase were 84.4%, 87.5%, 93.7%, and 93.8%, respectively. Specificity was 91.2% for leukocyte counts, 94.1% for alpha defensin, 97.0% for CRP, and 97.1% for leukocyte esterase. Diagnostic accuracy was 89.4% for alpha defensin, 92.4% for WBC counts and CRP, and 95.5% for leukocyte esterase. No statistical differences were observed in area under the curve (AUC) of the ROC curves of alpha defensin, CRP, and leukocyte counts. Logistic regression analysis applied to a model comprising all the variables showed an AUC higher than AUC of coupled variables. In conclusion, results of this study confirm the high sensitivity and specificity of synovial leukocyte esterase for diagnosis of prosthetic joint infection, also suggesting the need to assess a panel of markers to optimize diagnosis of these infections.

Epidemiology and Antibiotic Resistance of Late Prosthetic Knee and Hip Infections.

BACKGROUND: Prosthetic joint infections (PJI) are still a major complication of hip and knee arthroplasties. Identification of the causative pathogens and knowledge of their antibiotic susceptibilities are essential for the management of these infections. The main purpose of the study was to identify and compare the causative bacteria of prosthetic knee and hip joint infections in a reference Italian orthopedic center and to characterize antibiotic resistance profiles of bacteria involved. METHODS: Data from 429 patients with diagnosis of PJI were collected from January 2013 to June 2015: 229 presented a hip and 200 a knee prosthesis infection. Prostheses and periprosthetic tissues were treated with dithiothreitol before plating onto different media and broths. Identification and antimicrobial susceptibility testing were carried out by VITEK2 Compact (bioMerieux). RESULTS: There was not a substantial difference in the etiology of hip and knee PJI: staphylococci were the most frequently isolated bacteria in both groups, followed by Enterobacteriaceae and Propionibacterium acnes. Staphylococci showed a high rate of methicillin resistance (144 of 341) and a worrying frequency of isolates were resistant to teicoplanin (9%). Only 8.3% of Enterobacteriaceae produced extended-spectrum beta-lactamases, whereas the rate of carbapenemase-producing bacteria was not significant. CONCLUSION: We observed similar etiology of hip and knee PJIs. Nevertheless, bacteria isolated from knee showed higher resistance rates to glycopeptides and fluoroquinolones when compared with those isolated from the hip. The reason for this difference remains to be elucidated in future studies.

Osteonectin (SPARC) Expression in Vascular Calcification: In Vitro and Ex Vivo Studies.

Osteonectin, also termed SPARC, is a noncollagenous protein of bone matrix. Since there are controversial results regarding its role during the process of vascular calcification, we investigated osteonectin expression in our in vitro calcification model. Rat vascular smooth muscle cells (VSMCs) were challenged with high phosphate (5 mmol/L Pi) and analyzed quantifying calcium levels, through immunohistochemical studies, and studying gene expression. We detected a peak of osteonectin expression at day 7 in cell treated with high phosphate. The time course of calcium deposition, reflected the expression of osteonectin, resulting extensively present at day 7. On the contrary, the expression of the mitotic marker Ki-67 had a peak at day 4, showing no correlation between osteonectin and cell proliferation. Moreover, 7 days was the time point in which Cbfα1/RUNX-2 had its maximal expression. Furthermore, ascorbic acid increased osteonectin expression, supporting a procalcifying role for this protein. Next we decided to study osteonectin expression ex vivo in fetal, adult not calcified, and adult calcific vessels. Immunohistochemical studies demonstrated a spread and strong reactivity in VSMCs of a 20-week fetus, confirming that osteonectin may have a potential role in regulation of mitosis and in cell differentiation. In adult not calcified arteries, osteonectin was constitutively expressed and its levels increased in atherosclerotic and in calcified plaques, where it could have a regulatory role in the calcification process. Our in vitro and ex vivo data show osteonectin expression during the calcification process and suggest its potential role as procalcifying factor.

A new in vitro model to delay high phosphate-induced vascular calcification progression.

An increasing amount of patients affected by advanced chronic kidney disease suffer from vascular calcification (VC) that associates with cardiovascular morbidity and mortality. In this study, we created a new experimental in vitro model, trying to better elucidate high phosphate (Pi)-induced VC pathogenic mechanisms. Rat aortic vascular smooth muscle cells (VSMCs) were challenged for 7-10 days with high Pi with a repeated and short suspensions of high Pi treatment (intermittent suspension, IS) that was able to induce a significant inhibition of high Pi calcification, maximal at 5 h. Interestingly, the delay in calcification is a consequence of either the absence of free Pi or calcium-phosphate crystals being comparable to the total effect obtained during the 5 h-IS. The protective effect of IS was mediated by the reduction of apoptosis as demonstrated by the action of 20 µmol/L Z-VAD-FMK and by the preservation of the pro-survival receptor Axl expression. Furthermore, autophagy, during IS, was potentiated by increasing the autophagic flux, evaluated by LC3IIB western, while treating VSMCs with 1 mmol/L valproic acid did not affect VC. Finally, IS prevented VSMC osteoblastic differentiation by preserving smooth muscle lineage markers expression. Our data support the hypothesis that to delay significantly VC is necessary and sufficient the IS of high Pi challenge. The IS was able to prevent significantly apoptosis, to induce a potentiation in autophagy, and to prevent osteoblastic differentiation by preserving SM lineage markers.

Colonization by methicillin resistant staphylococci of nares and skin in healthcare workers: a pilot study in spinal surgeries.

INTRODUCTION: Staphylococcus aureus and coagulase-negative staphylococci (CoNS) colonization among healthcare workers (HCWs) may have implications in development of infections and in spreading of resistance. This study aimed to determine the rate of methicillin-resistant staphylococci carriage in HCWs of spinal surgeries in an Italian Orthopaedic Institute. MATERIALS AND METHODS: Samples from nares, axillae and hands were inoculated onto appropriate media in order to perform colony counts of methicillin-susceptible and resistant S. aureus and CoNS. RESULTS: Prevalence of S. aureus and CNS was 42.3% and 98%, respectively. Methicillin-resistance was rather infrequent in S. aureus (13.5%) while it was detected in most of CoNS (90.4%). Methicillin resistant S. aureus were prevalently isolated from nares while axillae showed the highest methicillin-resistant CoNS colonization rates. CONCLUSIONS: A relatively high rate of methicillin resistant staphylococci was found among HCWs in spinal surgeries wards, thus evidencing the need for careful prevention measures and for periodic evaluation of spread among HCWs.

Role and antimicrobial resistance of staphylococci involved in prosthetic joint infections.

PURPOSE: Staphylococci are responsible for approximately half of all prosthetic joint infections (PJIs) and they are often multi-drug resistant. The main purpose of this study was to evaluate the incidence of PJIs caused by staphylococci in our hospital from March 2010 to February 2012, with particular reference to antibiotic resistance in relation to their classification as contaminant or infecting isolates. METHODS: We analyzed samples recovered from 124 patients: most of them were male (55.8%) and the mean age was 66 ± 14 years. Prostheses derived from hip (54.8%) or knee (45.2%) replacement and they were processed by sonication. Isolates were identified using conventional biochemical methodologies. Antimicrobial susceptibility testing was carried out using the disk diffusion method as described by the European Committee on Antimicrobial Susceptibility Testing. RESULTS: A total of 135 staphylococci were isolated: the prevalent species was Staphylococcus aureus, but, on the whole, coagulase-negative staphylococci represented 57% of cases. Fifty-one isolates were recovered from a single sample and were therefore defined as contaminant. Linezolid and glycopeptides showed excellent activity versus all the tested isolates, while penicillin, levofloxacin, and erythromycin offered reduced activity against staphylococci. Interestingly, high resistance rates were observed for coagulase-negative staphylococci other than S. epidermidis classified as contaminant strains. CONCLUSIONS: We observed a remarkable spread of coagulase-negative staphylococci as causative agents of PJIs; but most of them were classified as contaminants. However, because of their low susceptibility to the antibiotics tested, further studies are necessary to evaluate their role as pathogens or as true contaminants.

Antiadhesive and antibiofilm activity of hyaluronic acid against bacteria responsible for respiratory tract infections.

To address the problem of limited efficacy of existing antibiotics in the treatment of bacterial biofilm, it is necessary to find alternative remedies. One candidate could be hyaluronic acid; this study therefore aimed to evaluate the in vitro antiadhesive and antibiofilm activity of hyaluronic acid toward bacterial species commonly isolated from respiratory infections. Interference exerted on bacterial adhesion was evaluated by using Hep-2 cells, while the antibiofilm activity was assessed by means of spectrophotometry after incubation of biofilm with hyaluronic acid and staining with crystal violet. Our data suggest that hyaluronic acid is able to interfere with bacterial adhesion to a cellular substrate in a concentration-dependent manner, being notably active when assessed as pure substance. Moreover, we found that Staphylococcus aureus biofilm was more sensitive to the action of hyaluronic acid than biofilm produced by Haemophilus influenzae and Moraxella catarrhalis. In conclusion, hyaluronic acid is characterized by notable antiadhesive properties, while it shows a moderate activity against bacterial biofilm. As bacterial adhesion to oral cells is the first step for colonization, these results further sustain the role of hyaluronic acid in prevention of respiratory infections.

Use of dithiothreitol to improve the diagnosis of prosthetic joint infections.

Diagnosis of prosthetic joint infections (PJIs) remains a challenge for microbiologists, despite new techniques for bacteria isolation have been developed in recent years. A widely recognized standard method has not yet been indicated mainly because of limitations due difficult procedures and need of dedicated instrumentation. We evaluated the ability of a sulfhydryl compound routinely used in microbiology laboratories, dithiothreitol (DTT), to dislodge bacteria from biofilm, keeping them alive and cultivable for identification and antibiotic susceptibility testing. We compared DTT treatment against sonication of prosthesis and culture of periprosthetic tissues, in order to establish if it could be introduced in routine microbiological diagnosis of PJIs. The study was conducted on 76 patients, 34 with aseptic loosening of their prosthesis and 42 who were diagnosed for PJI. DTT treatment gave results similar to sonication in terms of bacterial yielding. Sonication provided higher sensitivity (71.4%) and specificity (94.1%) respect to periprosthetic tissue culture, while DTT showed the same specificity of sonication but a better sensitivity (85.7%), especially when the causative microorganism was Staphylococcus epidermidis. In conclusion, we demonstrated that DTT could be used for PJIs diagnosis, thanks to its ease of use and its high sensitivity and specificity.

[Role of inflammation on renal anaemia]. / Ruolo dell'infiammazione nell'anemia renale.

It is well-known the central role of inflammation in the inhibition of erythropoiesis and iron availability in chronic kidney disease (CKD) patients with erythropoietin (EPO)-resistant anaemia. This inflammatory action is mediated by suppressive cytokines (i.e. IL-6, TNF-, INF-) inhibiting differentiation and proliferation activities of erythroid cells in the EPO-indipendent phase of erythropoiesis and stimulating hepcidin production for iron retention in reticulo-endothelial system and enterocytes. EPO resistance is associated with adverse outcomes, such as cardiovascular disease, faster progression to end stage renal disease and mortality. Treatment of the causes of EPO hyporesponsiveness including chronic inflammation results in an improvement of anaemia and a reduction in EPO requirements.