Prognostic Role of CSF β-amyloid 1-42/1-40 Ratio in Patients Affected by Amyotrophic Lateral Sclerosis

The involvement of β-amyloid (Aβ) in the pathogenesis of amyotrophic lateral sclerosis (ALS) has been widely discussed and its role in the disease is still a matter of debate. Aβ accumulates in the cortex and the anterior horn neurons of ALS patients and seems to affect their survival. To clarify the role of cerebrospinal fluid (CSF) Aβ 1-42 and Aβ 42/40 ratios as a potential prognostic biomarker for ALS, we performed a retrospective observational study on a cohort of ALS patients who underwent a lumbar puncture at the time of the diagnosis. CSF Aβ 1-40 and Aβ 1-42 ratios were detected by chemiluminescence immunoassay and their values were correlated with clinical features. We found a significant correlation of the Aβ 42/40 ratio with age at onset and Mini Mental State Examination (MMSE) scores. No significant correlation of Aβ 1-42 or Aβ 42/40 ratios to the rate of progression of the disease were found. Furthermore, when we stratified patients according to Aβ 1-42 concentration and the Aβ 42/40 ratio, we found that patients with a lower Aβ 42/40 ratio showed a shorter survival. Our results support the hypothesis that Aβ 1-42 could be involved in some pathogenic mechanism of ALS and we suggest the Aβ 42/40 ratio as a potential prognostic biomarker

Ataxin-1 and ataxin-2 intermediate-length PolyQ expansions in amyotrophic lateral sclerosis.

OBJECTIVE: Recent evidence suggests that intermediate-length polyglutamine (PolyQ) expansions in the ataxin-2 (ATXN-2) gene are a risk factor for amyotrophic lateral sclerosis (ALS). This work was undertaken with the aim to investigate the frequency of ataxin-1 (ATXN-1) and ATXN-2 PolyQ expansions in a cohort of patients with sporadic ALS (sALS) and patients with familial ALS (fALS) from southern Italy. METHODS: We assessed the PolyQ lengths of ATXN-1 and ATXN-2 in 405 patients with sALS, 13 patients with fALS, and 296 unrelated controls without history of neurodegenerative disorders. RESULTS: We found significantly higher intermediate PolyQ expansions ≥ 32 for ATXN-1 alleles and ≥ 28 for ATXN-2 alleles in the sALS cohort (ATXN-1: ALS, 7.07% vs controls, 2.38%; p = 0.0001; ATXN-2: ALS, 2.72% vs controls, 0.5%; p = 0.001). ATXN-1 CAT and ATXN-2 CAA interruptions were detected in patients with ALS only. Age at onset, site of onset, and sex were not significantly related to the ATXN-1 or ATXN-2 PolyQ repeat length expansions. CONCLUSIONS: Both ATXN-1 and ATXN-2 PolyQ intermediate expansions are independently associated with an increased risk for ALS

In hepatocellular carcinoma miR-221 modulates sorafenib resistance through inhibition of caspase-3\u2013mediated apoptosis

Purpose: The aberrant expression of miR-221 is a hallmark of human cancers, including hepatocellular carcinoma (HCC), and its involvement in drug resistance, together with a proved in vivo efficacy of anti-miR-221 molecules, strengthen its role as an attractive target candidate in the oncologic field. The discovery of biomarkers predicting the response to treatments represents a clinical challenge in the personalized treatment era. This study aimed to investigate the possible role of miR-221 as a circulating biomarker in HCC patients undergoing sorafenib treatment as well as to evaluate its contribution to sorafenib resistance in advanced HCC. Experimental Design: A chemically induced HCC rat model and a xenograft mouse model, together with HCC-derived cell lines were employed to analyze miR-221 modulation by Sorafenib treatment. Data from the functional analysis were validated in tissue samples from surgically resected HCCs. The variation of circulating miR-221 levels in relation to Sorafenib treatment were assayed in the animal models and in two independent cohorts of patients with advanced HCC. Results: MiR-221 over-expression was associated with Sorafenib resistance in two HCC animal models and caspase-3 was identified as its target gene, driving miR-221 anti-apoptotic activity following Sorafenib administration. Lower pre-treatment miR-221 serum levels were found in patients subsequently experiencing response to Sorafenib and an increase of circulating miR-221 at the two months assessment was observed in responder patients. Conclusions: MiR-221 might represent a candidate biomarker of likelihood of response to Sorafenib in HCC patients to be tested in future studies. Caspase-3 modulation by miR-221 participates to Sorafenib resistance

Dupilumab in the treatment of severe uncontrolled chronic rhinosinusitis with nasal polyps (CRSwNP): A multicentric observational Phase IV real-life study (DUPIREAL)

Background Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with significant morbidity and reduced health-related quality of life. Findings from clinical trials have demonstrated the effectiveness of dupilumab in CRSwNP, although real-world evidence is still limited. Methods This Phase IV real-life, observational, multicenter study assessed the effectiveness and safety of dupilumab in patients with severe uncontrolled CRSwNP (n = 648) over the first year of treatment. We collected data at baseline and after 1, 3, 6, 9, and 12 months of follow-up. We focused on nasal polyps score (NPS), symptoms, and olfactory function. We stratified outcomes by comorbidities, previous surgery, and adherence to intranasal corticosteroids, and examined the success rates based on current guidelines, as well as potential predictors of response at each timepoint. Results We observed a significant decrease in NPS from a median value of 6 (IQR 5–6) at baseline to 1.0 (IQR 0.0–2.0) at 12 months (p < .001), and a significant decrease in Sino-Nasal Outcomes Test-22 (SNOT-22) from a median score of 58 (IQR 49–70) at baseline to 11 (IQR 6–21; p < .001) at 12 months. Sniffin' Sticks scores showed a significant increase over 12 months (p < .001) compared to baseline. The results were unaffected by concomitant diseases, number of previous surgeries, and adherence to topical steroids, except for minor differences in rapidity of action. An excellent-moderate response was observed in 96.9% of patients at 12 months based on EPOS 2020 criteria. Conclusions Our findings from this large-scale real-life study support the effectiveness of dupilumab as an add-on therapy in patients with severe uncontrolled CRSwNP in reducing polyp size and improving the quality of life, severity of symptoms, nasal congestion, and smell

Association of Variants in the SPTLC1 Gene With Juvenile Amyotrophic Lateral Sclerosis

Importance: Juvenile amyotrophic lateral sclerosis (ALS) is a rare form of ALS characterized by age of symptom onset less than 25 years and a variable presentation.Objective: To identify the genetic variants associated with juvenile ALS.Design, Setting, and Participants: In this multicenter family-based genetic study, trio whole-exome sequencing was performed to identify the disease-associated gene in a case series of unrelated patients diagnosed with juvenile ALS and severe growth retardation. The patients and their family members were enrolled at academic hospitals and a government research facility between March 1, 2016, and March 13, 2020, and were observed until October 1, 2020. Whole-exome sequencing was also performed in a series of patients with juvenile ALS. A total of 66 patients with juvenile ALS and 6258 adult patients with ALS participated in the study. Patients were selected for the study based on their diagnosis, and all eligible participants were enrolled in the study. None of the participants had a family history of neurological disorders, suggesting de novo variants as the underlying genetic mechanism.Main Outcomes and Measures: De novo variants present only in the index case and not in unaffected family members.Results: Trio whole-exome sequencing was performed in 3 patients diagnosed with juvenile ALS and their parents. An additional 63 patients with juvenile ALS and 6258 adult patients with ALS were subsequently screened for variants in the SPTLC1 gene. De novo variants in SPTLC1 (p.Ala20Ser in 2 patients and p.Ser331Tyr in 1 patient) were identified in 3 unrelated patients diagnosed with juvenile ALS and failure to thrive. A fourth variant (p.Leu39del) was identified in a patient with juvenile ALS where parental DNA was unavailable. Variants in this gene have been previously shown to be associated with autosomal-dominant hereditary sensory autonomic neuropathy, type 1A, by disrupting an essential enzyme complex in the sphingolipid synthesis pathway.Conclusions and Relevance: These data broaden the phenotype associated with SPTLC1 and suggest that patients presenting with juvenile ALS should be screened for variants in this gene.</p

Genome-wide Analyses Identify KIF5A as a Novel ALS Gene

To identify novel genes associated with ALS, we undertook two lines of investigation. We carried out a genome-wide association study comparing 20,806 ALS cases and 59,804 controls. Independently, we performed a rare variant burden analysis comparing 1,138 index familial ALS cases and 19,494 controls. Through both approaches, we identified kinesin family member 5A (KIF5A) as a novel gene associated with ALS. Interestingly, mutations predominantly in the N-terminal motor domain of KIF5A are causative for two neurodegenerative diseases: hereditary spastic paraplegia (SPG10) and Charcot-Marie-Tooth type 2 (CMT2). In contrast, ALS-associated mutations are primarily located at the C-terminal cargo-binding tail domain and patients harboring loss-of-function mutations displayed an extended survival relative to typical ALS cases. Taken together, these results broaden the phenotype spectrum resulting from mutations in KIF5A and strengthen the role of cytoskeletal defects in the pathogenesis of ALS.Peer reviewe

Global disparities in surgeons’ workloads, academic engagement and rest periods: the on-calL shIft fOr geNEral SurgeonS (LIONESS) study

: The workload of general surgeons is multifaceted, encompassing not only surgical procedures but also a myriad of other responsibilities. From April to May 2023, we conducted a CHERRIES-compliant internet-based survey analyzing clinical practice, academic engagement, and post-on-call rest. The questionnaire featured six sections with 35 questions. Statistical analysis used Chi-square tests, ANOVA, and logistic regression (SPSS® v. 28). The survey received a total of 1.046 responses (65.4%). Over 78.0% of responders came from Europe, 65.1% came from a general surgery unit; 92.8% of European and 87.5% of North American respondents were involved in research, compared to 71.7% in Africa. Europe led in publishing research studies (6.6 ± 8.6 yearly). Teaching involvement was high in North America (100%) and Africa (91.7%). Surgeons reported an average of 6.7 ± 4.9 on-call shifts per month, with European and North American surgeons experiencing 6.5 ± 4.9 and 7.8 ± 4.1 on-calls monthly, respectively. African surgeons had the highest on-call frequency (8.7 ± 6.1). Post-on-call, only 35.1% of respondents received a day off. Europeans were most likely (40%) to have a day off, while African surgeons were least likely (6.7%). On the adjusted multivariable analysis HDI (Human Development Index) (aOR 1.993) hospital capacity &gt; 400 beds (aOR 2.423), working in a specialty surgery unit (aOR 2.087), and making the on-call in-house (aOR 5.446), significantly predicted the likelihood of having a day off after an on-call shift. Our study revealed critical insights into the disparities in workload, access to research, and professional opportunities for surgeons across different continents, underscored by the HDI

Le virus adéno-associé dans le foie : histoire naturelle de l'infection et conséquences sur le développement des tumeurs

Le virus adéno-associé (AAV) est un virus à ADN monobrin, défectif et endémique dans la population humaine. L'infection à AAV a longtemps été considérée comme non pathogénique. Cependant, il y a quelques années, nous avons identifié pour la première fois des insertions clonales récurrentes d'AAV2 impliquées dans la pathogenèse du carcinome hépatocellulaire humain (CHC) développé sur un foie normal. Ces insertions virales clonales ciblent des oncogènes et entraînent leur surexpression. À ce jour, peu de travaux ont étudié l'infection à AAV de type sauvage dans le foie humain. Dans ce travail, nous avons étudié l'histoire naturelle de l'infection virale dans les tissus hépatiques et ses conséquences sur le développement de tumeurs dans une vaste cohorte de patients (n = 1464). La présence de l'AAV est observée chez 21% des patients, plus fréquemment dans la contrepartie non-tumorale (18%) que dans la tumeur (8%) et significativement enrichie chez les femmes, les patients jeunes et non cirrhotiques. Deux sous-types d’AAV ont été identifiés dans le foie, l’AAV2 classique et un génotype hybride AAV2-AAV3-AAV13, avec une fréquence égale dans notre cohorte. Nous avons détecté la présence de formes épisomales d'AAV dans 27% des tissus non tumoraux positifs pour l'AAV, associée de manière significative à l'expression de l'ARN viral et à la co-infection par des virus auxiliaires, suggérant une infection active en cours. Nous avons identifié le virus de l'herpès humain de type 6 (HHV6) comme étant le virus auxiliaire naturel de l'AAV dans le foie. En revanche, l'ADN de l'adénovirus n'a été détecté que chez 0,5% des patients et aucune association avec l'AAV n'a été constatée. Nous avons confirmé la sélection positive des insertions d'AAV clonales au cours du développement du CHC chez les patients sans cirrhose dans 2% des tumeurs ciblant CCNA2, CCNE1, TERT, TNFSF10, KMT2B et INHBE / GLI1. De plus, les altérations de CCNA2 et de CCNE1 dues à des insertions virales d'AAV et de VHB ou à des réarrangements structurels ont défini une nouvelle sous-classe de CHC (CCN-HCC) et un nouveau mécanisme de développement du CHC sur le foie normal, améliorant nos connaissances sur l'hépatocarcinogénèse sur le foie non cirrhotique. Les CCN-HCC présentent également des caractéristiques moléculaires particulières qui pourraient être ciblées par un traitement spécifique.Adeno-associated virus (AAV) is a defective mono-stranded DNA virus, endemic in human population. AAV infection has long been considered as non-pathogenic, however few years ago we reported for the first time recurrent clonal AAV2 insertion in the pathogenesis of human hepatocellular carcinoma (HCC) developed on normal liver. These clonal viral insertions target cancer driver genes leading to their overexpression. To date, little is known about wild type AAV infection in human liver. In this work we investigated the natural history of the viral infection in the liver tissues and the consequences in tumor development in a large cohort of patients (n=1464). The presence of AAV was observed in 21% of patients, more frequently in the non-tumor counterpart (18%) than in tumor (8%) and significantly enriched in young, female and non-cirrhotic patients. Two AAV subtypes were identified in the liver, the classical AAV2 and a hybrid AAV2-AAV3-AAV13 genotypes, with an equal frequency in our cohort. We detected the presence of episomal AAV forms in 27% of AAV positive non-tumor tissues significantly associated with viral RNA expression and co-infection with helper viruses suggesting an ongoing active infection. We identified human herpes virus type 6 (HHV6) as the natural AAV helper virus in the liver. In contrast, adenovirus DNA was detected in only 0.5% of patients and no association with AAV was found. We confirmed the positive selection of clonal AAV insertions during HCC development in patients without cirrhosis in 2% of tumors targeting CCNA2, CCNE1, TERT, TNFSF10, KMT2B and INHBE/GLI1. Moreover, the alterations in CCNA2 and CCNE1 due to viral insertions of AAV and HBV or structural rearrangements defined a new subclass of HCCs (CCN-HCC) and a novel mechanism of HCC development on normal liver improving our knowledge on hepatocarcinogenesis on non-cirrhotic liver. CCN-HCCs display also peculiar molecular features that could be targeted by specific treatment

URIC ACID LEVELS IN SERUM AND CSF OF ALS PATIENTS

Objective: Urate (UA) is a potent antioxidant that effectively scavenges reactive nitrogen and oxygen radicals, and persons with a high plasma UA level may be at lower risk of some neurodegenerative disorders, as Parkinson’s disease (PD). Low plasma UA level has been observed in Alzheimer’s disease (AD) and vascular dementia, but there is no data on correlations to neuropsychological test results in these patient groups. Amyotrophic Lateral Sclerosis (ALS) is a devastating motor neuron disease, with a highly variable rate of progression and whose diagnosis is chiefly based on clinical and neurophysiological parameters. The etiopathogenesis is unknown, but the oxidative stress seems to play an important role. As UA works as an antioxidant, and given the phenotypic heterogeneity of the disorder (in terms of site of onset and clinical progression), we asked whether UA could represent a biological marker in ALS. Therefore, in the present study we assayed UA in the CSF and serum of ALS patients and disease controls, and evaluated its relationship with different clinical variables and the rate of progression. Patients: 146 ALS patients (M/F 1.8; mean age at onset 61± 11.65 years; 41 with bulbar onset, 35%) who met the Revised El Escorial criteria for clinically definite, probable or probable with laboratory support were selected. General demographic and clinical features of this cohort were recorded. Controls were 141 patients admitted in our hospital for suspected neurological disorders such as multiple sclerosis, myelitis and polyneuropathy. CSF and serum from ALS patients and disease controls was obtained as a necessary step of the diagnostic work-up and all patients gave an informed consent to the procedure. Methods: CSF and serum were obtained from fasted patients and aliquots stored at –80°C until use. UA level in CSF and serum was assaying using a colorimeter enzymatic test (UA plus, COBAS ®). Results: The median serum level of UA (mg/dl) was 4.65 (3.6-5.6) in the ALS patients and 4.80 (3.9-5.6) in controls (p=ns). Further, UA CSF levels (mg/dl) were no significantly different between ALS and controls (ALS, 0.3 [0.2-0.4] vs controls, 0.28 [0.2-0.4], p=ns). CSF and serum UA in ALS was nor related to the severity of the disease (as measured as a rate of progression with the clinimetric Appel ALS rating scale), respiratory function, age at onset and site of onset. Conclusion: UA is not a biochemical marker in ALS and is probably unrelated to its pathophysiolog

Tau protein as a diagnostic and prognostic biomarker in Amyotrophic Lateral Sclerosis

BACKGROUND: To test the hypothesis that total tau (tTau), tau phosphorylated at threonine 181 (pTau) and the pTau/tTau ratio in the cerebrospinal fluid (CSF) are diagnostic and prognostic biomarkers of amyotrophic lateral sclerosis (ALS), we performed a retrospective observational study on a large cohort of ALS patients and controls.METHODS: We enrolled 196 ALS patients and 91 controls, including ALS-mimics and patients with non-neurodegenerative diseases. All patients underwent lumbar puncture for CSF analysis at the time of the diagnostic work-up or to first referral. We measured tTau and pTau levels in the CSF by chemiluminescence enzyme immunoassay (CLEIA).RESULTS: ALS patients showed significantly higher levels of CSF tTau and lower pTau/tTau ratio than controls (tTau: 245 vs 146 pg/mL; p&lt;0.001; pTau/tTau ratio: 0.12 vs 0.18; p&lt;0.001, respectively). No differences of pTau levels were detected. The analysis of the receiver operating characteristic (ROC) curve showed a good diagnostic accuracy of tTau and pTau/tTau ratio (tTau: AUC 0.685, 95%CI 0.616-0.754, p=0.039; pTau/tTau ratio: AUC 0.777, 95%CI 0.707-0.848, p&lt;0.001). Among ALS patients, increased tTau levels were associated with the advanced age of onset, increased DeltaFS, rate of progression, and spinal onset. The multivariate analysis showed that in ALS patients, this biomarker was an independent negative predictor of overall survival.CONCLUSIONS: Our findings suggest that tTau and pTau/tTau ratio can be diagnostic biomarkers of ALS. Finally, CSF tTau level at diagnosis might play a relevant prognostic role in the disease