Wet-dry-wet drug screen leads to the synthesis of TS1, a novel compound reversing lung fibrosis through inhibition of myofibroblast differentiation

Therapies halting the progression of fibrosis are ineffective and limited. Activated myofibroblasts are emerging as important targets in the progression of fibrotic diseases. Previously, we performed a high-throughput screen on lung fibroblasts and subsequently demonstrated that the inhibition of myofibroblast activation is able to prevent lung fibrosis in bleomycin-treated mice. High-throughput screens are an ideal method of repurposing drugs, yet they contain an intrinsic limitation, which is the size of the library itself. Here, we exploited the data from our “wet” screen and used “dry” machine learning analysis to virtually screen millions of compounds, identifying novel anti-fibrotic hits which target myofibroblast differentiation, many of which were structurally related to dopamine. We synthesized and validated several compounds ex vivo (“wet”) and confirmed that both dopamine and its derivative TS1 are powerful inhibitors of myofibroblast activation. We further used RNAi-mediated knock-down and demonstrated that both molecules act through the dopamine receptor 3 and exert their anti-fibrotic effect by inhibiting the canonical transforming growth factor β pathway. Furthermore, molecular modelling confirmed the capability of TS1 to bind both human and mouse dopamine receptor 3. The anti-fibrotic effect on human cells was confirmed using primary fibroblasts from idiopathic pulmonary fibrosis patients. Finally, TS1 prevented and reversed disease progression in a murine model of lung fibrosis. Both our interdisciplinary approach and our novel compound TS1 are promising tools for understanding and combating lung fibrosis

Endovascular Abdominal Aortic Aneurysm Repair With Ovation Alto Stent Graft: Protocol for the ALTAIR (ALTo endogrAft Italian Registry) Study

Background: Since 2010, the Ovation Abdominal Stent Graft System has offered an innovative sealing option for abdominal aortic aneurysm (AAA) by including a sealing ring filled with polymer 13 mm from the renal arteries. In August 2020, the redesigned Ovation Alto, with a sealing ring 6 mm closer to the top of the fabric, received CE Mark approval. Objective: This registry study aims to evaluate intraoperative, perioperative, and postoperative results in patients treated by the Alto stent graft (Endologix Inc.) for elective AAA repair in a multicentric consecutive experience. Methods: All consecutive eligible patients submitted to endovascular aneurysm repair (EVAR) by Alto Endovascular AAA implantation will be included in this analysis. Patients will be submitted to EVAR procedures based on their own preferences, anatomical features, and operators experience. An estimated number of 300 patients submitted to EVAR with Alto stent graft should be enrolled. It is estimated that the inclusion period will be 24 months. The follow-up period is set to be 5 years. Full data sets and cross-sectional images of contrast-enhanced computed tomography scan performed before EVAR, at the first postoperative month, at 24 or 36 months, and at 5-year follow-up interval will be reported in the central database for a centralized core laboratory review of morphological changes. The primary endpoint of the study is to evaluate the technical and clinical success of EVAR with the Alto stent graft in short- (90-day), mid- (1-year), and long-term (5-year) follow-up periods. The following secondary endpoints will be also addressed: operative time; intraoperative radiation exposure; contrast medium usage; AAA sac shrinkage at 12-month and 5-year follow-up; any potential role of patients' baseline characteristics, valuated on preoperative computed tomography angiographic study, and of device configuration (number of component) in the primary endpoint. Results: The study is currently in the recruitment phase and the final patient is expected to be treated by the end of 2023 and then followed up for 5 years. A total of 300 patients will be recruited. Analyses will focus on primary and secondary endpoints. Updated results will be shared at 1- and 3-5-year follow-ups. Conclusions: The results from this registry study could validate the safety and effectiveness of the new design of the Ovation Alto Stent Graft. The technical modifications to the endograft could allow for accommodation of a more comprehensive range of anatomies on-label

Targeted Approach to Distinguish and Determine Absolute Levels of GDF8 and GDF11 in Mouse Serum

Growth differentiation factor 11 (GDF11) is a TGF-\u3b2 superfamily circulating factor that regulates cardiomyocyte size in rodents, sharing 90% amino acid sequence identity in the active domains with myostatin (GDF8)\u2014the major determinant of skeletal muscle mass. Conflicting data on age-related changes in circulating levels have been reported mainly due to the lack of specific detection methods. More recently, liquid chromatography tandem mass spectrometry (LC-MS/MS) based assay showed that the circulating levels of GDF11 do not change significantly throughout human lifespan, but GDF8 levels decrease with aging in men. Here a novel detection method is demonstrated based on parallel reaction monitoring LC-MS/MS assay combined with immunoprecipitation to reliably distinguish GDF11 and GDF8 as well as determine their endogenous levels in mouse serum. The data indicate that both GDF11 and GDF8 circulating levels significantly decline with aging in female mice

ETDRS-fast: implementing psychophysical adaptive methods to standardized visual acuity measurement with ETDRS charts

PURPOSE. To measure visual acuity (VA) on Early Treatment Diabetic Retinopathy Study (ETDRS) charts with a modified faster procedure (ETDRS-Fast), based on adaptive psychophys- ics methods and to assess the method’s validity and reproduc- ibility. METHODS. Whereas the standard method for measuring VA with the ETDRS charts requires that the subject read all the letters beginning with the top row, in the ETDRS-Fast procedure, the subject is asked to read only one letter per row until a mistake is made. Then, following simple rules, the examiner finds a row from which the subject can begin reading all the letters down- ward, thus making the method identical with the standard method near threshold. VA determination was performed twice with both methods in 57 subjects in two separate ses- sions to assess validity and reproducibility. RESULTS. In both sessions the correlation between the two procedures was high (intraclass correlation coefficient 0.95), confirming the validity of the ETDRS-Fast procedure. Repro- ducibility was good for both procedures, with intraclass cor- relation coefficients of 0.94 for the standard and 0.96 for the ETDRS-Fast method. The ETDRS-Fast procedure allowed a sig- nificantly shorter test duration ( 30%; P 0.0001). CONCLUSIONS. Adaptive procedures allow accurate and fast de- termination of psychophysical thresholds by reducing the number of stimulus presentations when the subject is far from threshold. In the ETDRS-Fast method a few simple rules ap- plied to optotype chart reading allow adaptation to each pa- tient’s level of VA. The ETDRS-Fast procedure significantly reduces test time and still yields results that are as accurate as those obtained with the standard method

Retroillumination versus reflected-light images in the photographic assessment of posterior capsule opacification

PURPOSE. To investigate the relative merit of retroillumination and of reflected light slit-lamp– derived photographs in the assessment of the opacification of the posterior lens capsule. METHODS. Retroillumination and slit-lamp–derived reflected-light photographs were taken on 23 consecutive eyes with posterior capsule opacification (PCO) in uncomplicated pseudophakia. Subjective grading was performed on both types of photographs to evaluate the extent and density of posterior capsular opacification. Best-corrected visual acuity (BCVA) before and after YAG laser capsulotomy was used to assess the impact of capsular opacification on visual function. RESULTS. After capsulotomy all patients attained a BCVA 46 letters ( 20/32) with a mean increase of 25 letters, indicating that PCO was the cause of visual impairment in these patients. The relative capacity of retroillumination and of reflected-light photographs to adequately capture the extent and the severity of posterior capsule opacification varied considerably. Reflected-light images, in addition to frequently producing higher severity scores for the opacity than retroillumination photographs, in 4 of 23 eyes (17.4%) proved to be the only technique able to document the presence of PCO. CONCLUSIONS. Our results indicate that, with respect to retroillumination images, reflected-light photography has an increased ability to adequately capture the presence and the severity of PCO and that the use of only retroillumination images may lead to its underestimation. This may be relevant to clinical studies aiming to evaluate incidence and progression of this condition

Targeted Approach to Distinguish and Determine Absolute Levels of GDF8 and GDF11 in Mouse Serum

Growth differentiation factor 11 (GDF11) is a TGF-β superfamily circulating factor that regulates cardiomyocyte size in rodents, sharing 90% amino acid sequence identity in the active domains with myostatin (GDF8)—the major determinant of skeletal muscle mass. Conflicting data on age-related changes in circulating levels have been reported mainly due to the lack of specific detection methods. More recently, liquid chromatography tandem mass spectrometry (LC-MS/MS) based assay showed that the circulating levels of GDF11 do not change significantly throughout human lifespan, but GDF8 levels decrease with aging in men. Here a novel detection method is demonstrated based on parallel reaction monitoring LC-MS/MS assay combined with immunoprecipitation to reliably distinguish GDF11 and GDF8 as well as determine their endogenous levels in mouse serum. The data indicate that both GDF11 and GDF8 circulating levels significantly decline with aging in female mice

Profile and progression of cognitive deficits in Progressive Supranuclear Palsy, Multiple System Atrophy and Parkinson's Disease

Objective: This study investigated neuropsychological tests most sensitive in differentiating progressive supranuclear palsy (PSP) from Parkinson’s disease (PD) and multiple system atrophy (MSA), and in detecting cognitive changes at follow-up. Background: Cognitive impairment is frequent in PSP, but its characteristics and progression need to be properly defined. Method: We evaluated 35 PSP with Richardson’s syndrome (PSP-RS), 30 MSA as well as 65 age-, sex-, and education-matched PD with an extensive clinical and neuropsychological assessment, allowing Level II cognitive diagnosis. Eighteen PSP, 12 MSA and 30 PD had a second evaluation 12-18-month (mean 15 months) after the first assessment. Results: In PSP, Montreal Cognitive Assessment (MoCA), verbal fluencies (phonemic and semantic tasks), Stroop test, Digit Span Sequencing (DSS), incomplete letters of Visual Object and Space Perception (VOSP) and Benton’s Judgment of Line Orientation (JLO) performance were significantly impaired at baseline compared to PD and MSA. Executive and visuo-spatial abilities declined longitudinally in PSP, but not in PD and MSA. After 1.5 year, 16% of PSP converted to dementia. Conclusion: Our study provides evidence that cognitive decline is more severe and rapid in PSP than PD and MSA. MoCA, verbal fluency, DSS and Benton’s JLO are valuable tests to detect cognitive progression in PSP and may be proposed as biomarker for research protocols to assess efficacy of disease modification strategies