Explore the vast range of titles available.

Background Patients with Parkinsonian syndromes, including Parkinson’s disease (PD) and atypical parkinsonism (AP), are at increased risk of disability following hip fracture. Limited data are available on their functional recovery after inpatient rehabilitation. This study aimed to compare rehabilitation outcomes in patients with parkinsonism versus non-parkinsonian controls after hip fracture surgery and to identify clinical predictors of worse recovery within the parkinsonian group. Methods A retrospective study was conducted on 60 patients with parkinsonism (45 PD, 15 AP) and 60 age- and sex-matched non-parkinsonian controls undergoing inpatient rehabilitation after hip fracture surgery. Functional status was assessed at admission and discharge using the Modified Barthel Index (MBI) and modified Rankin Scale (mRS). Linear mixed-effects models were applied to explore associations between clinical variables (including diagnosis, dementia, dysphagia, postural instability) and outcomes. Results Patients with parkinsonism had significantly lower functional scores at both admission and discharge as compared to controls ( p  < 0.001), with AP patients performing worse than PD patients ( p  = 0.02). Among parkinsonian individuals, dementia, dysphagia, and postural instability were independently associated with worse outcomes ( p  < 0.001). Although MBI change scores (delta-MBI) did not differ significantly between groups, mRS change (delta-mRS) was greater in controls, suggesting that parkinsonian patients experienced a higher overall disability burden at both admission and discharge, despite a smaller relative improvement. These findings indicate that non-motor symptoms substantially influence rehabilitation trajectories and should be considered in planning care. Conclusions Parkinsonian patients, particularly those with atypical parkinsonism, experience worse functional outcomes after hip fracture rehabilitation when compared with non-parkinsonian individuals. The presence of dementia, dysphagia, and postural instability further impairs recovery.

BackgroundThe epidemiology of aortic stenosis (AS) in older patients admitted to non-cardiologic acute hospital wards and the effect of AS on mid-term survival are incompletely reported. In a cohort of very old patients admitted to an acute geriatric unit (AGU), we aimed to assess: (1) the prevalence of newly and previously diagnosed AS; and (2) the association between AS severity and patients’ 6-month mortality.MethodsThe patients consecutively admitted in two AGU rooms from February 2016 to February 2018 were assessed with echocardiography and AS severity was defined according to standard criteria. We assessed frailty using a 34-item Frailty Index (34-FI), which was operationalized using health variable information, and the Clinical Frailty Scale (CFS). Vital status at 6 months was extracted from Regional Register of Birth and Death.ResultsTwo hundred and three patients (mean age 84.5 ± 6.0 SD, female gender 56.1%) were included. Of these, 57 (28.1%) had AS, mild in 9 (4.5%), moderate in 32 (16.1%) and severe in 16 (8.1%). A new diagnosis of AS was obtained in 42 (73.7%) patients, of whom 33 (78.6%) had moderate or severe AS. At 6 months, 61 (28.9%) patients died. In multiple regression models, after adjusting for covariates, frailty, as assessed with both FI and CFS, was independent predictor of 6-month mortality whereas AS was not.ConclusionsAmong older patients admitted to non-cardiologic acute hospital wards, AS was common and frequently underdiagnosed. The severity of AS was not associated with increased 6-month mortality, whereas frailty was the most important predictor.

Biomolecular condensates serve as membrane-less compartments within cells, concentrating proteins and nucleic acids to facilitate precise spatial and temporal orchestration of various biological processes. The diversity of these processes and the substantial variability in condensate characteristics present a formidable challenge for quantifying their molecular dynamics, surpassing the capabilities of conventional microscopy. Here, we show that our single-photon microscope provides a comprehensive live-cell spectroscopy and imaging framework for investigating biomolecular condensation. Leveraging a single-photon detector array, single-photon microscopy enhances the potential of quantitative confocal microscopy by providing access to fluorescence signals at the single-photon level. Our platform incorporates photon spatiotemporal tagging, which allowed us to perform time-lapse super-resolved imaging for molecular sub-diffraction environment organization with simultaneous monitoring of molecular mobility, interactions, and nano-environment properties through fluorescence lifetime fluctuation spectroscopy. This integrated correlative study reveals the dynamics and interactions of RNA-binding proteins involved in forming stress granules, a specific type of biomolecular condensates, across a wide range of spatial and temporal scales. Our versatile framework opens up avenues for exploring a broad spectrum of biomolecular processes beyond the formation of membrane-less organelles. The wide variety of cellular processes involving biomolecular condensation makes their quantification a challenging task. Here, the authors present an integrated platform based on single-photon microscopy to study complex biomolecular processes.

Fluorescence laser-scanning microscopy (LSM) is experiencing a revolution thanks to new single-photon (SP) array detectors, which give access to an entirely new set of single-photon information. Together with the blooming of new SP LSM techniques and the development of tailored SP array detectors, there is a growing need for (i) DAQ systems capable of handling the high-throughput and high-resolution photon information generated by these detectors, and (ii) incorporating these DAQ protocols in existing fluorescence LSMs. We developed an open-source, low-cost, multi-channel time-tagging module (TTM) based on a field-programmable gate array that can tag in parallel multiple single-photon events, with 30 ps precision, and multiple synchronisation events, with 4 ns precision. We use the TTM to demonstrate live-cell super-resolved fluorescence lifetime image scanning microscopy and fluorescence lifetime fluctuation spectroscopy. We expect that our BrightEyes-TTM will support the microscopy community in spreading SP-LSM in many life science laboratories. The authors developed an open-source, low-cost, multi-channel time-tagging module for fluorescence lifetime image scanning microscopy and correlation spectroscopy that can tag in parallel multiple single-photon events with 30 ps precision.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease due to gradual motoneurons (MN) degeneration. Among the processes associated to ALS pathogenesis, there is the formation of cytoplasmic inclusions produced by aggregation of mutant proteins, among which the RNA binding protein FUS. Here we show that, in neuronal cells and in iPSC-derived MN expressing mutant FUS, such inclusions are significantly reduced in number and dissolve faster when the RNA m 6 A content is diminished. Interestingly, stress granules formed in ALS conditions showed a distinctive transcriptome with respect to control cells, which reverted to similar to control after m 6 A downregulation. Notably, cells expressing mutant FUS were characterized by higher m 6 A levels suggesting a possible link between m 6 A homeostasis and pathological aggregates. Finally, we show that FUS inclusions are reduced also in patient-derived fibroblasts treated with STM-2457, an inhibitor of METTL3 activity, paving the way for its possible use for counteracting aggregate formation in ALS. In Amyotrophic Lateral Sclerosis (ALS), formation of cytoplasmic inclusions by mutant protein aggregation is observed. Here the authors show that these inclusions dissolve faster when m 6 A RNA modification is inhibited in ALS cellular models.

Background Skeletal muscle mass wasting almost invariably accompanies bone loss in elderly, and the coexistence of these two conditions depends on the tight endocrine crosstalk existing between the two organs, other than the biomechanical coupling. Since the current diagnostics limitation in this field, and given the progressive population aging, more effective tools are needed. The aim of this study was to identify circulating microRNAs (miRNAs) as potential biomarkers for muscle mass wasting in post‐menopausal osteoporotic women. Methods One hundred seventy‐nine miRNAs were assayed by quantitative real‐time polymerase chain reaction in plasma samples from 28 otherwise healthy post‐menopausal osteoporotic women (73.4 ± 6.6 years old). The cohort was divided in tertiles based on appendicular skeletal muscle mass index (ASMMI) to better highlight the differences on skeletal muscle mass (first tertile: n = 9, ASMMI = 4.88 ± 0.40 kg·m−2; second tertile: n = 10, ASMMI = 5.73 ± 0.23 kg·m−2; third tertile: n = 9, ASMMI = 6.40 ± 0.22 kg·m−2). Receiver operating characteristic (ROC) curves were calculated to estimate the diagnostic potential of miRNAs. miRNAs displaying a statistically significant fold change ≥ ±1.5 and area under the curve (AUC) > 0.800 (P < 0.05) between the first and third tertiles were considered. A linear regression model was applied to estimate the association between miRNA expression and ASMMI in the whole population, adjusting for body mass index, age, total fat (measured by total‐body dual‐energy X‐ray absorptiometry [DXA]) and bone mineral density (measured by femur DXA). Circulating levels of adipo‐myokines were evaluated by bead‐based immunofluorescent assays and enzyme‐linked immunosorbent assays. Results Five miRNAs (hsa‐miR‐221‐3p, hsa‐miR‐374b‐5p, hsa‐miR‐146a‐5p, hsa‐miR‐126‐5p and hsa‐miR‐425‐5p) resulted down‐regulated and two miRNAs (hsa‐miR‐145‐5p and hsa‐miR‐25‐3p) were up‐regulated in the first tertile (relative‐low ASMMI) compared with the third tertile (relative‐high ASMMI) (fold change ≥ ±1.5; P‐value < 0.05). All the corresponding ROC curves had AUC > 0.8 (P < 0.05). Two signatures hsa‐miR‐126‐5p, hsa‐miR‐146a‐5p and hsa‐miR‐425‐5p; and hsa‐miR‐126‐5p, hsa‐miR‐146a‐5p, hsa‐miR‐145‐5p and hsa‐miR‐25‐3p showed the highest AUC, 0.914 (sensitivity = 77.78%; specificity = 100.00%) and 0.901 (sensitivity = 88.89%; specificity = 100.00%), respectively. Conclusions In this study, we identified, for the first time, two miRNA signatures, hsa‐miR‐126‐5p, hsa‐miR‐146a‐5p and hsa‐miR‐425‐5p; and hsa‐miR‐126‐5p, hsa‐miR‐146a‐5p, hsa‐miR‐145‐5p and hsa‐miR‐25‐3p, specifically associated with muscle mass wasting in post‐menopausal osteoporotic women.

G-quadruplexes (G4s) are four-stranded DNA or RNA structures formed by guanine-rich sequences. They occur in functional regions of the genomic material, including the promoter part of genes, regulatory region, and telomeric threads. G4s play a key role in various biological processes, including transcription, replication, and telomere maintenance. Guanidine-containing derivatives can bind to G-quadruplexes, either by intercalating into the structure or by interacting with the grooves or loops. The binding can stabilize the G-quadruplex, potentially affecting its biological function. In this paper, the ability of guanidinium-containing hordatines to interact with G4 was evaluated. Analogues lacking the guanidinium group or showing the benzofuran system instead of the dihydrobenzofuran core were prepared and tested as well. NMR titration and docking calculations were used to probe the binding of the compounds to G4 of c-MYC oncogene. Spectroscopic analyses were consistent with a significant interaction of benzofurans 3 and 4 at the 5′-end and 3′-end tetrads and with the formation of ligand/G-quadruplex complexes with a 2:1 stoichiometry. The resulting data were supported by docking simulations. Cytotoxic activity was evaluated on a model of U2OS osteosarcoma (ATCC HTB-96) and breast cancer (MDA-MB-231) cell lines, further highlighting the key role of the guanidinium fragment and the benzofuran core in the G-quadruplex stabilization.

G-quadruplexes are secondary structures originating from nucleic acid regions rich in guanines, which are well known for their involvement in gene transcription and regulation and DNA damage repair. In recent studies from our group, kynurenic acid (KYNA) derivative 1 was synthesized and found to share the structural features typical of G-quadruplex binders. Herein, structural modifications were conducted on this scaffold in order to assist the binding with a G-quadruplex, by introducing charged hydrophilic groups. The antiproliferative activity of the new analogues was evaluated on an IGROV-1 human ovarian cancer cell line, and the most active compound, compound 9, was analyzed with NMR spectrometry in order to investigate its binding mode with DNA. The results indicated that a weak, non-specific interaction was set with duplex nucleotides; on the other hand, titration in the presence of a G-quadruplex from human telomere d(TTAGGGT)4 showed a stable, although not strong, interaction at the 3′-end of the nucleotidic sequence, efficiently assisted by salt bridges between the quaternary nitrogen and the external phosphate groups. Overall, this work can be considered a platform for the development of a new class of potential G-quadruplex stabilizing molecules, confirming the crucial role of a planar system and the ability of charged nitrogen-containing groups to facilitate the binding to G-quadruplex grooves and loops.

Image Scanning Microscopy (ISM) enables good signal-to-noise ratio (SNR), super-resolution and high information content imaging by leveraging array detection in a laser-scanning architecture. However, the SNR is still limited by the size of the detector, which is conventionally small to avoid collecting out-of-focus light. Nonetheless, the ISM dataset inherently contains the axial information of the fluorescence emitters. We leverage this knowledge to achieve computational optical sectioning without sacrificing the conventional benefits of ISM. We invert the physical model to fuse the raw dataset into a single image with improved sampling, SNR. lateral resolution, and optical sectioning. We provide a complete theoretical framework and validate our approach with experimental images of biological samples acquired with a custom setup equipped with a single photon avalanche diode (SPAD) array detector. Furthermore, we generalize our method to other imaging techniques, such as multi-photon excitation fluorescence microscopy and fluoresce lifetime imaging. To enable this latter, we take advantage of the single-photon timing ability of SPAD arrays, accessing additional sample information. Our method outperforms conventional reconstruction techniques and opens new perspectives for exploring the unique spatio-temporal information provided by SPAD array detectors.

Image Scanning Microscopy (ISM) enables good signal-to-noise ratio (SNR), super-resolution and high information content imaging by leveraging array detection in a laser-scanning architecture. However, the SNR is still limited by the size of the detector, which is conventionally small to avoid collecting out-of-focus light. Nonetheless, the ISM dataset inherently contains the axial information of the fluorescence emitters. We leverage this knowledge to achieve computational optical sectioning without sacrificing the conventional benefits of ISM. We invert the physical model to fuse the raw dataset into a single image with improved sampling, SNR. lateral resolution, and optical sectioning. We provide a complete theoretical framework and validate our approach with experimental images of biological samples acquired with a custom setup equipped with a single photon avalanche diode (SPAD) array detector. Furthermore, we generalize our method to other imaging techniques, such as multi-photon excitation fluorescence microscopy and fluoresce lifetime imaging. To enable this latter, we take advantage of the single-photon timing ability of SPAD arrays, accessing additional sample information. Our method outperforms conventional reconstruction techniques and opens new perspectives for exploring the unique spatio-temporal information provided by SPAD array detectors.