Direct-acting antivirals used in HCV-related liver disease do not affect thyroid function and autoimmunity.

PURPOSE: It is well known that interferon-α (IFN-α), used for long time as the main therapy for HCV-related disease, induces thyroid alterations, but the impact of the new direct-acting antivirals (DAAs) on thyroid is not established. Aim of this prospective study was to evaluate if DAAs therapy may induce thyroid alterations. METHODS: A total of 113 HCV patients, subdivided at the time of the enrollment in naïve group (n = 64) and in IFN-α group (n = 49) previously treated with pegylated interferon-α and ribavirin, were evaluated for thyroid function and autoimmunity before and after 20-32 weeks of DAAs. RESULTS: Before starting DAAs, a total of 8/113 (7.1%) patients showed Hashimoto's thyroiditis (HT) all belonging to IFN-α group (8/49, 16.3%), while no HT cases were found in the naïve group. Overall, 7/113 (6.2%) patients were hypothyroid: 3/64 (4.7%) belonging to naïve group and 4/49 (8.2%) to IFN-α group. Furthermore, a total of 8/113 patients (7.1%) showed subclinical hyperthyroidism: 2/64 (3.1%) were from naïve group and 6/49 (12.2%) from IFN-α group. Interestingly, after DAAs therapy, no new cases of HT, hypothyroidism and hyperthyroidism was found in all series, while 6/11 (54.5%) patients with non-autoimmune subclinical thyroid dysfunction became euthyroid. Finally, the only association between viral genotypes and thyroid alterations was genotype 1 and hypothyroidism. CONCLUSIONS: This study supports evidence that DAAs have a limited or missing influence on thyroid in patients with HCV-related diseases. Moreover, it provides preliminary evidence that subclinical non-autoimmune thyroid dysfunction may improve after HCV infection resolution obtained by DAAs.

Exploring potential mechanisms for zinc deficiency to impact in autism spectrum disorder: a narrative review.

Autism spectrum disorder (ASD) is a heterogeneous and complex group of life-long neurodevelopmental disorders. How this clinical condition impacts an individual's intellectual, social and emotional capacities, contributing to alterations in the proprioceptive and sensory systems and increasing their selective attitude towards food, is well described in the literature. This complex condition or status exposes individuals with ASD to an increased risk of developing overweight, obesity and non-communicable diseases compared with the neurotypical population. Moreover, individuals with ASD are characterised by higher levels of inflammation, oxidative stress markers and intestinal dysbiosis. All these clinical features may also appear in zinc deficiency (ZD) condition. In fact, zinc is an essential micronutrient for human health, serving as a structural, catalytic and regulatory component in numerous physiological processes. The aim of this narrative review is to explore role of ZD in ASD. Factors affecting zinc absorption, excretion and dietary intake in this vulnerable population are taken into consideration. Starting from this manuscript, the authors encourage future research to investigate the role of ZD in ASD. The perspective is to potentially find another missing piece in the 'ASD clinical puzzle picture' to improve the health status of these individuals.

The influence of mandibular stretching on athletes subjected to high intensity workout.

The use of oral appliances is considered effective in improving sport performance. Jaw clenching or stretching, improved breathing or positioning of the jaw have been proposed to explain the improvement. In this study, we reported the effects of regularly practiced mandibular stretching at submaximal opening of the jaw in Crossfit athletes. Seventeen test subjects undergoing mandibular stretching (MS) showed significantly reduced (p0.001) performance time in a high-intensity workout measured before and after seven days of 10-minutes stretching practiced twice daily, while no improvement was observed in control subjects who did not perform the stretching. Additionally, blood lactate levels appeared to recover more quickly in MS subjects.Crossfit is a type of training regimen that pushes the limits of bodily recovery processes. Jaw stretching, eliciting the trigeminocardiac reflex (a reflex decrease in heart rate and blood pressure following trigeminal stimulation) and central oxygen-conserving mechanisms could help relieve post-training stress and improve training recovery.

Detection of environmental contamination with feline and canine parvoviruses: new perspectives and challenges.

AIMS: To develop a protocol for environmental sampling to detect parvoviruses of dogs and cats in the environment. METHODS AND RESULTS: Environmental contamination was carried out using different dilutions of parvovirus-contaminated materials; further field samplings were performed in areas in which clinical cases of parvovirus infections were present. Sterile cotton swabs and sponges for microbial surface sampling were used. Viruses were detected in these samples with different methods: conventional PCR, nested PCR and real-time PCR, detecting viral DNA; virus isolation, detecting infectious virus; and a commercial rapid enzyme immunoassay, detecting viral antigen. No substantial differences were observed in the two sampling methods, although the sponge was more convenient for sampling rough surfaces. Molecular assays were the most sensitive methods, identifying even very low amounts of viral DNA (up to 10 copies of viral DNA/10 µl of sample). Virus isolation and the rapid test detected the viruses only at the highest viral concentrations, both in the experimental setting and field conditions. CONCLUSIONS: Environmental sampling and molecular protocols were effective in detecting environmental contamination with parvoviruses. SIGNIFICANCE AND IMPACT OF THE STUDY: The protocol will be useful to identify possible sources of infection and to assess the efficacy of disinfection protocols in the environment.

Learning effect and repeatability of stabilometric measurements: "standard" vs. usual foot placement.

The existence of a learning effect by which subjects progressively reduce body sway over the course of repetitive stabilometric measurements is currently debated. Also, the position and orientation of the feet on the platform can have a substantial influence on the outcome measurements. The aim of the present work was to assess the effect of feet positions on mean total velocity (V) of the center of pressure and the area (AR) covered by its displacements during quiet standing. A group of 35 healthy young subjects was examined during two successive sessions consisting of five recordings with their feet placed either in the recommended (standard, SP) or their usual most comfortable (UP) position. Results show a slight decreasing trend that failed to be statistically significant checked with Friedman's ANOVA (SP AR, χ2(4)=6.10, p=0.19 and V, χ2(4)=8.66, p=0.07 and UP AR, χ2(4)=2.32, p=0.68 and V, χ2(4)=1.19, p=0.88). Nonetheless, values of AR and V showed a notable decrement especially evident in the SP exam reaching, respectively, 24% and 11% from baseline, whereas variability measured by the coefficient of variation was the same in the two exams. Given the results, a learning effect should not be ruled out with confidence. Also, usual foot placement would be preferable to avoid this effect. Further research is needed to take into consideration the great variability of stabilometric measurements and the fact that different subjects could adapt more readily to the test conditions than others.

Arthroscopic transosseous versus suture anchor repair: clinical outcomes in patients with bilateral rotator cuff tears.

The aim of our study was to define if Arthroscopic Transosseous Rotator Cuff Techniques should have comparable results to those of the suture-anchors technique in a single row configuration. We reported the preliminary results of a consecutive population of 22 patients who underwent a rotator cuff treatment on the left and right sides for average medium-sized thickness tears with minimal fatty infiltration with the two different techniques: transosseous rotator cuff repair technique on one side and single row with suture-anchors on the other side, in different times. Subjective evaluation with DASH questionnaires, Constant Scores and Numerical Rating Scale (NRS) for pain evaluation, have been submitted pre and postoperatively after both operations. A statistical analysis was performed to assess the superiority of one technique and to compare pre and postoperative ROM data and clinical outcomes. A transosseous rotator cuff repair was performed in 7 patients on the dominant arm, while the other 15 patients had dominant arm cuff tear lesions repaired by using suture-anchors technique. At last follow-up a significant improvement, in shoulder pain and function, was referred at both sides. Also, DASH, Constant Scores and NRS for pain evaluation improved with both techniques, but no statistical difference was found between them. Arthroscopic transosseous rotator cuff repair technique shows comparable results to those of the suture-anchors technique in a single row configuration.

Changes of sensory and pain thresholds in healthy subjects after mandibular extension at maximum mouth opening: implications for temporomandibular disorders therapy.

There is evidence showing that increasing the vertical dimension of occlusion, or interocclusal distance, reduces pain symptoms in patients with temporomandibular disorders (TMD) and bruxism. The mechanism underlying the analgesic effect is still under investigation. Some researchers propose either a gate mechanism or the activation of the trigemino-cardiac reflex. In this study, sensory and pain thresholds changes in the mandibular district have been evaluated in twenty healthy subjects immediately after 5 minutes of mandibular stretching at maximum opening of the jaw. Results showed a 60% increment in sensory threshold and a 70% increment in pain threshold (p<0.001) compared with the baseline values. The magnitude of the analgesic effect seems proportional to the extent of mandibular opening. These and similar results are discussed as for the implication in the treatment of TMD related symptoms.

Postural Disorders Produced by School Furniture on a Population of a Junior High School.

In recent years people suffering of backache has significantly increased. This led us to focus our attention on the adequacy of school furniture for the correct development of the adult posture. The standing posture of 67 students of middle school has been analyzed at the beginning and at the end of two consecutive scholastic years using a stabilometric platform. Starting from the second year, about half of the students were provided with the furniture designed following European standard (UNI EN 1729:2006), while the other half maintained the traditional one. The main purpose of this research has been to verify by means of posturometric analysis the effects on postural parameters of the use of traditional furniture in comparison to the furniture following the UNI EN 1729:2006. We observed that prolonged sitting at school changes some posturometric variables of schoolchildren. Since no differences has been found between the two groups, the validity of the European standards is questioned. The present study allowed us to single out four anthropometric parameters that should be considered in order to devise a new model of adjustable furniture. By adjusting every year the furniture of each student, it would be possible to avoid (at least at school) the adoption of wrong postural positions that could be responsible for backache and other common musculoskeletal disorders.

An observational study of the effects of using an high oral splint on pain control

Increasing occlusal vertical dimension (OVD) by means of oral splints is a practice widely used in the treatment of temporomandibular disorders (TMD), specifically myofascial pain, although the results are still uncertain. Oral splints with a vertical height that significantly exceeds the clinical rest position are considered by some researchers to be a better therapeutic solution in alleviating TMD symptoms than are "low" splints. In our observational study, 21 patients suffering from myofascial pain were examined for the effects of wearing a "high" oral splint while sleeping for 3 months. To ensure proper splint making, a mandibular stretching procedure was used to induce a relaxation of the patients' masticatory muscles and allow the correct alignment of the jaws. Results showed a marked increase of the interocclusal distance or "free space" (hence of OVD; from 0.64±0.53 mm to 1.42±0.76 mm, p<0.0001) measured by a kinesiograph, followed by a substantial reduction of the intensity of pain in oral and extraoral regions after using the splint. These results support the view that increasing OVD beyond the clinical rest position is not detrimental to patients' health. More importantly, high oral splints has been shown to be a promising therapeutic aid for the treatment of TMD and correlated pain syndromes. This clinical trial was registered on clinicaltrials.gov (Identifier: NCT02908568).

Plasma trough concentrations of darunavir/ritonavir and raltegravir in older patients with HIV-1 infection.

OBJECTIVES: The aim of the study was to assess plasma concentrations of darunavir/ritonavir and raltegravir in older patients compared with younger patients with HIV-1 infection. METHODS: In this observational, open-label study, adult HIV-infected out-patients aged ≤ 40 years (younger patients) or ≥ 60 years (older patients) and treated with tenofovir/emtricitabine plus darunavir/ritonavir (800/100 mg daily) or raltegravir (400 mg twice daily) were asked to participate. The trough concentrations (Ctrough ) of darunavir/ritonavir and raltegravir were assessed at steady state using a validated high-performance liquid chromatography (HPLC)-tandem mass spectrometry method. RESULTS: A total of 88 HIV-positive patients were enrolled in the study. Forty-six patients were treated with darunavir/ritonavir, and 42 with raltegravir. The geometric mean plasma Ctrough (coefficient of variation) of raltegravir was comparable between the 19 older and 23 younger subjects: 106 ng/mL (151%) and 94 ng/mL (129%), respectively [geometric mean ratio (GMR) 0.85; 95% confidence interval (CI) 0.71-1.57; P = 0.087]. In contrast, the geometric mean plasma Ctrough of darunavir was significantly higher among the 21 older patients [2209 ng/mL (139%)] than among the 25 younger patients [1876 ng/mL (162%); GMR 1.56; 95% CI: 1.22-1.88; P = 0.004]. Similarly, the geometric mean Ctrough of ritonavir was significantly higher among older than among younger individuals. CONCLUSIONS: The mean plasma Ctrough of darunavir and ritonavir was significantly higher in older patients than in younger patients with HIV-1 infection, while the mean plasma level of raltegravir was comparable in the two groups. However, both regimens showed good tolerability in both younger and older subjects.

Extensibility and Distensibility of the Thoracic Aorta in Patients with Aneurysm.

OBJECTIVES: Reference values of aortic deformation during the cardiac cycle can be valuable for the pre-operative planning of thoracic endovascular aortic repair (TEVAR) and for facilitating computational fluid dynamics. This study aimed to quantify normal aortic extensibility (longitudinal extension) and distensibility (radial expansion), as well as pulsatile strain, in a group of 10 (>60 years) individuals with abdominal or thoracic aortic aneurysms. METHODS: ECG gated CT images of the thoracic aorta were reconstructed into virtual 3D models of aortic geometry. The centre lumen line length of the thoracic aorta and three longitudinal segments, and the aortic diameter and luminal areas of four radial intersections were extracted with a dedicated software script to calculate extensibility, longitudinal strain, distensibility, and circumferential area strain. RESULTS: Mean extensibility and longitudinal strain of the entire thoracic aorta were 3.5 [1.3-6.8] × 10-3 N-1, and 2.7 [1.0-4.5]%, respectively. Extensibility and longitudinal strain were most pronounced in the ascending aorta (20.6 [5.7-36.2] × 10-3 N-1 and 15.9 [6.6-31.9]%) and smallest in the descending aorta (4.4 [1.6-12.3] × 10-3 N-1 and 2.2 [0.7-4.7]%). Mean distensibility and circumferential area strain were most pronounced at the sinotubular junction (1.7 [0.5-2.9] × 10-3 mmHg-1 and 11.3 [3.3-18.5]%, respectively). Distensibility varied between 0.9 [0.3-2.5] × 10-3 mmHg-1 and 1.2 [0.3-3.3] × 10-3 mmHg-1 at the intersections in the aortic arch and descending aorta. CONCLUSIONS: Pulsatile deformations in both longitudinal and circumferential directions are considerable throughout the thoracic aorta. These findings may have implications for pre-operative TEVAR planning and highlight the need for devices that can mimic the significant aortic longitudinal and circumferential strains.

In vivo estimation of the shoulder joint center of rotation using magneto-inertial sensors: MRI-based accuracy and repeatability assessment.

BACKGROUND: The human gleno-humeral joint is normally represented as a spherical hinge and its center of rotation is used to construct humerus anatomical axes and as reduction point for the computation of the internal joint moments. The position of the gleno-humeral joint center (GHJC) can be estimated by recording ad hoc shoulder joint movement following a functional approach. In the last years, extensive research has been conducted to improve GHJC estimate as obtained from positioning systems such as stereo-photogrammetry or electromagnetic tracking. Conversely, despite the growing interest for wearable technologies in the field of human movement analysis, no studies investigated the problem of GHJC estimation using miniaturized magneto-inertial measurement units (MIMUs). The aim of this study was to evaluate both accuracy and precision of the GHJC estimation as obtained using a MIMU-based methodology and a functional approach. METHODS: Five different functional methods were implemented and comparatively assessed under different experimental conditions (two types of shoulder motions: cross and star type motion; two joint velocities: ωmax = 90°/s, 180°/s; two ranges of motion: Ɵ = 45°, 90°). Validation was conducted on five healthy subjects and true GHJC locations were obtained using magnetic resonance imaging. RESULTS: The best performing methods (NAP and SAC) showed an accuracy in the estimate of the GHJC between 20.6 and 21.9 mm and repeatability values between 9.4 and 10.4 mm. Methods performance did not show significant differences for the type of arm motion analyzed or a reduction of the arm angular velocity (180°/s and 90°/s). In addition, a reduction of the joint range of motion (90° and 45°) did not seem to influence significantly the GHJC position estimate except in a few subject-method combinations. CONCLUSIONS: MIMU-based functional methods can be used to estimate the GHJC position in vivo with errors of the same order of magnitude than those obtained using traditionally stereo-photogrammetric techniques. The methodology proposed seemed to be robust under different experimental conditions. The present paper was awarded as "SIAMOC Best Methodological Paper 2016".

MicroRNAs Association in the Cardiac Hypertrophy Secondary to Complex Congenital Heart Disease in Children.

Complex congenital heart disease (CHD) affects cardiac blood flow, generating a pressure overload in the compromised ventricles and provoking hypertrophy that over time will induce myocardial dysfunction and cause a potential risk of imminent death. Therefore, the early diagnosis of complex CHD is paramount during the first year of life, with surgical treatment of patients favoring survival. In the present study, we analyzed cardiac tissue and plasma of children with cardiac hypertrophy (CH) secondary to CHD for the expression of 11 miRNAs specific to CH in adults. The results were compared with the miRNA expression patterns in tissue and blood of healthy children. In this way, we determined that miRNAs 1, 18b, 21, 23b, 133a, 195, and 208b constitute the expression profile of the cardiac tissue of children with CHD. Meanwhile, miRNAs 21, 23a, 23b, and 24 can be considered specific biomarkers for the diagnosis of CH in infants with CHD. These results suggest that CH secondary to CHD in children differs in its mechanism from that described for adult hypertrophy, offering a new perspective to study the development of this pathology and to determine the potential of hypertrophic miRNAs to be biomarkers for early CH.

Comparison of brainstem reflex recordings and evoked potentials with clinical and MRI data to assess brainstem dysfunction in multiple sclerosis: a short-term follow-up.

Brainstem dysfunctions are associated to high risk of developing severe disability in patients with multiple sclerosis (PwMS), often undetected by conventional routine assessments. In this view, the purpose of this study was to monitor brainstem function over a short-term period in PwMS, comparing clinical and magnetic resonance imaging (MRI) examinations with evoked potentials (EPs) and brainstem reflexes (BSRs). Forty-five PwMS were evaluated at baseline and after 15.1 ± 4.2 months through Expanded Disability Status Scale (EDSS) score, MRI, EPs, vestibulo-masseteric (VMR), acoustic-masseteric (AMR), vestibulo-collic (VCR) and trigemino-collic (TCR) reflexes. At baseline, brainstem alterations were detected by EDSS, MRI, EPs and BSRs in 40, 77.8, 84.4 and 82.2 % of patients, respectively. At follow-up, EDSS and MRI remained unchanged, while EP and BSR deteriorated in 86.7 and 91.1 % of patients, respectively. Changes from 1 to 3 altered EPs and from 1 to 4 altered BSRs were significant only for EPs (p = 0.028). The analysis of grading severity for each test disclosed significant worsening of the VMR, AMR, TCR and P14 wave of the median somatosensory EP. Combined EP/BSR recordings were significantly more sensitive than paired EDSS/MRI assessments at baseline (93.3 versus 80 %; p = 0.006) and follow-up (97.8 versus 82.2 %; p = 0.008). In the short-term VMR, AMR, TCR and P14 wave disclosed a significant functional brainstem deterioration by detecting lesions that remained clinically and MRI silent. Our findings provide evidence for a valuable role of neurophysiological methods, especially BSRs, in investigating and monitoring brainstem dysfunctions in MS, in comparison with the standard clinical and MRI procedures.

Treatment of chronic pulmonary aspergillosis with voriconazole: review of a case series.

PURPOSE: Chronic pulmonary aspergillosis (CPA) is a rare disease that primarily affects subjects with moderate immunodepression and/or structural alterations in the lung. METHODS: Data for patients with probable CPA were collected over 24 months. Patients with probable CPA received oral voriconazole, and clinical, laboratory and radiological follow-up was performed at 3, 6 and 12 months. RESULTS: 21 patients (mean age 52.4 years) were evaluated. Factors predisposing to CPA were tuberculosis (n = 8), chronic obstructive pulmonary disease (n = 7), corticosteroids (n = 14), chemo- or radio-therapy (n = 6), tracheostomy or endotracheal prosthesis (n = 5), smoking (n = 4), asthma (n = 3), and chronic liver disease (n = 3). Sputum or bronchial aspirate cultures were positive for Aspergillus spp. in 14 cases (66.6 %). (1,3)-ß-D-glucan on serum was positive in 16 cases (76.2 %). Excavated pulmonary thickening was evident in 14 patients (66.6 %) and in 9 of these cases (64.2 %) aspergilloma was present. [(18)F]2-fluoro-2-deoxy-D-glucose-PET-CT was positive in 13/15 patients, and simple aspergilloma was diagnosed after surgical excision in one of the negative cases. All patients were treated with oral voriconazole. Therapy was discontinued due to skin toxicity (n = 3), liver toxicity (n = 2) and severe mental disorder (n = 1). At 12 months' follow-up, nine patients (42.9 %) were considered cured or improved. Seven patients (33.3 %) died during follow-up, mainly due to underlying disease. CONCLUSIONS: A reasonable proportion of patients achieved cure or improvement with voriconazole, but 28.5 % of treated patients had to discontinue therapy because of toxicity. The high mortality makes it difficult to fully assess the real efficacy of voriconazole and to establish the correct duration of therapy.

Actin and myosin contribute to mammalian mitochondrial DNA maintenance.

Mitochondrial DNA maintenance and segregation are dependent on the actin cytoskeleton in budding yeast. We found two cytoskeletal proteins among six proteins tightly associated with rat liver mitochondrial DNA: non-muscle myosin heavy chain IIA and ß-actin. In human cells, transient gene silencing of MYH9 (encoding non-muscle myosin heavy chain IIA), or the closely related MYH10 gene (encoding non-muscle myosin heavy chain IIB), altered the topology and increased the copy number of mitochondrial DNA; and the latter effect was enhanced when both genes were targeted simultaneously. In contrast, genetic ablation of non-muscle myosin IIB was associated with a 60% decrease in mitochondrial DNA copy number in mouse embryonic fibroblasts, compared to control cells. Gene silencing of ß-actin also affected mitochondrial DNA copy number and organization. Protease-protection experiments and iodixanol gradient analysis suggest some ß-actin and non-muscle myosin heavy chain IIA reside within human mitochondria and confirm that they are associated with mitochondrial DNA. Collectively, these results strongly implicate the actomyosin cytoskeleton in mammalian mitochondrial DNA maintenance.

Impact of the maximum ring difference on image quality and noise characteristics of a total-body PET/CT scanner.

The sensitivity of a PET system highly depends on the axial acceptance angle or maximum ring difference (MRD), which can be particularly high for total-body scanners due to their larger axial field of views (aFOVs). This study aims to evaluate the impact on image quality (IQ) and noise performance when MRD85 (18°), the current standard for clinical use, is increased to MRD322 (52°) for the Biograph Vision Quadra (Siemens Healthineers). METHODS: Studies with a cylindrical phantom covering the 106 cm aFOV and an IEC phantom filled with 18F, 68Ga and 89Zr were performed for acquisition times from 60 to 1800 s and activity concentrations from 0.4 to 3 kBq/ml to assess uniformity, contrast recovery coefficients (CRCs) and to characterize noise by coefficient of variation (CV). Spatial resolution was compared for both MRDs by sampling a quadrant of the FOV with a point source. Further IQ, CV, liver SUVmean and SUVmax were compared for a cohort of 5 patients scanned with [18F]FDG (3 MBq/kg, 1 h p.i.) from 30 to 300 s. RESULTS: CV was improved by a factor of up to 1.49 and is highest for short acquisition times, peaks at the center field of view and mitigates parabolic in axial direction with no difference to MRD85 beyond the central 80 cm. No substantial differences between the two evaluated MRDs in regards to uniformity, SUVmean or CRC for the different isotopes were observed. A degradation of the average spatial resolution of 0.9 ±â€¯0.2 mm in the central 40 cm FOV was determined with MRD322. Depending on the acquisition time MRD322 resulted in a decrease of SUVmax between 23.8% (30 s) and 9.0% (300 s). CONCLUSION: Patient and phantom studies revealed that scan time could be lowered by approximately a factor of two with MRD322 while maintaining similar noise performance. The moderate degradation in spatial resolution for MRD322 is worth to exploit the full potential of the Quadra by either shorten scan times or leverage noise performance in particular for low count scenarios such as ultra-late imaging or dynamic studies with high temporal resolution.

Genetic diversity and its impact on disease severity in respiratory syncytial virus subtype-A and -B bronchiolitis before and after pandemic restrictions in Rome.

OBJECTIVES: To scrutinize whether the high circulation of respiratory syncytial virus (RSV) observed in 2021-2022 and 2022-2023 was due to viral diversity, we characterized RSV-A and -B strains causing bronchiolitis in Rome, before and after the COVID-19 pandemic. METHODS: RSV-positive samples, prospectively collected from infants hospitalized for bronchiolitis from 2017-2018 to 2022-2023, were sequenced in the G gene; phylogenetic results and amino acid substitutions were analyzed. Subtype-specific data were compared among seasons. RESULTS: Predominance of RSV-A and -B alternated in the pre-pandemic seasons; RSV-A dominated in 2021-2022 whereas RSV-B was predominant in 2022-2023. RSV-A sequences were ON1 genotype but quite distant from the ancestor; two divergent clades included sequences from pre- and post-pandemic seasons. Nearly all RSV-B were BA10 genotype; a divergent clade included only strains from 2021-2022 to 2022-2023. RSV-A cases had lower need of O2 therapy and of intensive care during 2021-2022 with respect to all other seasons. RSV-B infected infants were more frequently admitted to intensive care units and needed O2 in 2022-2023. CONCLUSIONS: The intense RSV peak in 2021-2022, driven by RSV-A phylogenetically related to pre-pandemic strains is attributable to the immune debt created by pandemic restrictions. The RSV-B genetic divergence observed in post-pandemic strains may have increased the RSV-B specific immune debt, being a possible contributor to bronchiolitis severity in 2022-2023.

Unraveling the complexity of anti-doping analysis: reassessing meldonium detection and doping verdicts in a case study.

BACKGROUND: The precision and accuracy of mass spectrometry (MS) made it a fundamental tool in anti-doping analysis. High-resolution (HR) mass spectrometers significantly improved compound identification. This study systematically analyzes data from an athlete (Subject 1) who tested positive for meldonium and compares it with data from a healthy volunteer (Subject 2) to examine the correctness of the doping verdict. CASE PRESENTATION: The documentation related to Subject 1 was thoroughly processed and analyzed. A study involving a volunteer (Subject 2) replicated Subject 1 regimen and urine sample collection for data alignment with anti-doping results, with Subject 2 reporting not using meldonium. The anti-doping agency's analysis of Subject 1 showed the presence of meldonium at a concentration close to the established cut-off level. However, a closer examination revealed that one specific ion, crucial for meldonium identification, was absent from the mass spectra. Analyzing Subject 2 data, using the same methodology, the absence of the specific ion was confirmed, even though the volunteer did not consume meldonium. The European directive and the method that was validated and cited by the anti-doping agency identified meldonium on at least four specific ions, whereas the anti-doping analysis used only three ions. This discrepancy compromises the specificity of meldonium identification. CONCLUSIONS: To enhance the analytical methodology, two strategic interventions are suggested: adjusting the meldonium cut-off value and expanding the analysis to include meldonium metabolites. By addressing these avenues, the precision of meldonium detection and doping verdicts can be improved. In conclusion, this study challenges the anti-doping agency's verdict and prompts a reevaluation of meldonium detection methodologies in anti-doping measures.