Valeria Piazza

Hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC) is a recently described tubulinopathy caused by mutations in the tubulin beta-4a isotype. It is a severe disorder characterized by progressive myelin loss and neurological impairment. The taiep rat, a spontaneous TUBB4A mutant, is a model for investigating the cellular and molecular mechanisms underlying this disease. Homozygous taiep rats exhibit hallmark features of H-ABC, including early-onset hypomyelination, progressive demyelination, and microtubule accumulation in oligodendrocytes, whereas heterozygous animals appear to maintain normal myelin structure using conventional techniques. We combined hyperspectral imaging (HSI) and Nile Red (NR) staining to differentiate homozygous taiep and wild-type animals and determine whether subtle myelin alterations could be detected in heterozygous rats. Remarkably, heterozygous animals also showed changes in the Nile Red emission profile, with fluorescence shifted toward longer wavelengths compared with controls, uncovering previously unrecognized modifications in the myelin microenvironment. Using spectral phasor analysis, these alterations were localized and quantified without model assumptions, demonstrating the sensitivity of this approach for detecting subtle pathological changes. Together, our findings contribute to the characterization of myelin-associated spectral changes in this tubulinopathy and support the use of spectral microscopy to distinguish alterations in myelin properties, with potential applications in the study of other white matter disorders.

Background: Patients with implantable cardioverter defibrillators (ICDs) or cardiac resynchronization therapy with defibrillator (CRT-D) can be followed-up with remote monitoring(RM), which allows the evaluation of complex and serious arrhythmias .Objectives: The aim of this study was to evaluate the prognostic value of ventricular arrhythmia alerts on mortality in patients with implantable devicesMethods: A retrospective cohort study was conducted in 62 patients, analyzing demographic and clinical data and RM alerts: non-sustained ventricular tachycardia (NSVT), ventricular tachycardia (VT), or ventricular fibrillation (VF)and its prognostic value for mortality .Results: 35.5% of patients presented alerts for ventricular arrhythmias. They had worse left ventricular ejection fraction and a higher prevalence of heart failure. During follow-up, they had higher all-cause mortality (40.9% in the group with alerts vs. 7.5% in the group without alerts; p=0.003). Survival analysis confirmed that these patients had a significantly lower probability of survival (Log-Rank test, p=0.006).Conclusion: The occurrence of alerts for ventricular arrhythmias detected by RM was a significant prognostic marker of lower survival, underscoring the value of this technology for risk stratification and clinical decision-making. A larger patient cohort and longer follow-up are needed to confirm its independent prognostic value.

The taiep rat is a tubulin mutant with an early hypomyelination followed by progressive demyelination of the central nervous system due to a point mutation in the Tubb4a gene. It shows clinical, radiological, and pathological signs like those of the human leukodystrophy hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC). Taiep rats had tremor, ataxia, immobility episodes, epilepsy, and paralysis; the acronym of these signs given the name to this autosomal recessive trait. The aim of this study was to analyze the characteristics of somatosensory evoked potentials (SSEPs) and motor evoked potentials (MEPs) in adult taiep rats and in a patient suffering from H-ABC. Additionally, we evaluated the effects of 4-aminopyridine (4-AP) on sensory responses and locomotion and finally, we compared myelin loss in the spinal cord of adult taiep and wild type (WT) rats using immunostaining. Our results showed delayed SSEPs in the upper and the absence of them in the lower extremities in a human patient. In taiep rats SSEPs had a delayed second negative evoked responses and were more susceptible to delayed responses with iterative stimulation with respect to WT. MEPs were produced by bipolar stimulation of the primary motor cortex generating a direct wave in WT rats followed by several indirect waves, but taiep rats had fused MEPs. Importantly, taiep SSEPs improved after systemic administration of 4-AP, a potassium channel blocker, and this drug induced an increase in the horizontal displacement measured in a novelty-induced locomotor test. In taiep subjects have a significant decrease in the immunostaining of myelin in the anterior and ventral funiculi of the lumbar spinal cord with respect to WT rats. In conclusion, evoked potentials are useful to evaluate myelin alterations in a leukodystrophy, which improved after systemic administration of 4-AP. Our results have a translational value because our findings have implications in future medical trials for H-ABC patients or with other leukodystrophies.

Cysteine-rich receptor-like kinases (CRKs) are a type of receptor-like kinases (RLKs) that are important for pathogen resistance, extracellular reactive oxygen species (ROS) signaling, and programmed cell death in plants. In a previous study, we identified 46 CRK family members in the Phaseolus vulgaris genome and found that CRK12 was highly upregulated under root nodule symbiotic conditions. To better understand the role of CRK12 in the Phaseolus–Rhizobia symbiotic interaction, we functionally characterized this gene by overexpressing (CRK12-OE) and silencing (CRK12-RNAi) it in a P. vulgaris hairy root system. We found that the constitutive expression of CRK12 led to an increase in root hair length and the expression of root hair regulatory genes, while silencing the gene had the opposite effect. During symbiosis, CRK12-RNAi resulted in a significant reduction in nodule numbers, while CRK12-OE roots showed a dramatic increase in rhizobial infection threads and the number of nodules. Nodule cross sections revealed that silenced nodules had very few infected cells, while CRK12-OE nodules had enlarged infected cells, whose numbers had increased compared to controls. As expected, CRK12-RNAi negatively affected nitrogen fixation, while CRK12-OE nodules fixed 1.5 times more nitrogen than controls. Expression levels of genes involved in symbiosis and ROS signaling, as well as nitrogen export genes, supported the nodule phenotypes. Moreover, nodule senescence was prolonged in CRK12-overexpressing roots. Subcellular localization assays showed that the PvCRK12 protein localized to the plasma membrane, and the spatiotemporal expression patterns of the CRK12-promoter::GUS-GFP analysis revealed a symbiosis-specific expression of CRK12 during the early stages of rhizobial infection and in the development of nodules. Our findings suggest that CRK12, a membrane RLK, is a novel regulator of Phaseolus vulgaris-Rhizobium tropici symbiosis.

The simultaneous surface and internal measurements from a chemically modified cortical bovine bone suffering a plastic range deformation are presented. Since the bone is an anisotropic structure, its mechanical response could be modified if its organic or inorganic phases change. The latter could result in high plastic deformations, where the interferometrical signal from an optical analysis is easily de-correlated. In this work, digital holography interferometry (DHI) and Fourier domain optical coherence tomography (FD-OCT) are used to analyze the plastic range deformation of the bone under compression. The simultaneous use of these two optical methods gives information even when one of them de-correlates. The surface results retrieved with DHI show the high anisotropy of the bone as a continuously increasing displacement field map. Meanwhile, the internal information obtained with FD-OCT records larger deformations at different depths. Due to the optical phase, it is possible to complement the measurements of these two methods during the plastic deformation. (c) 2023 Optica Publishing Group

The satisfactory visualization of cytoskeletal components in the brain is challenging. The ubiquitous distribution of the networks of microtubules, microfilaments, and intermediate filaments in all the neural tissues, together with the variability in the outcomes of fluorescent protein fusion strategies and their limited applicability to dynamic studies of antibodies and drugs as chromophore vehicles, make classical optical approaches not as effective as for other proteins. When tubulin needs to be studied, the label-free generation of second harmonics is a very suitable option due to the non-centrosymmetric organization of the molecule. This technique, when conjugated to microscopy, can qualitatively describe the volumetric distribution of parallel bundles of microtubules in biological samples, with the additional advantage of working with fresh tissues that are unfixed and unpermeabilized. This work describes how to image tubulin with a commercial second harmonic generation microscopy setup to highlight microtubules in the tubulin-enriched structures of the oligodendrocytes, as in hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC) tubulinopathy, a recently described myelin disorder.

Hypomyelination with atrophy of the basal ganglia and cerebellum is a recently described tubulinopathy caused by a mutation in the tubulin beta 4a isoform, expressed in oligodendrocytes. The taiep rat is the only spontaneous tubulin beta 4a mutant available for the study of this pathology. We aimed to identify the effects of the tubulin mutation on freshly collected, unstained samples of the central white matter of taiep rats using second harmonic generation microscopy. Cytoskeletal differences between the central white matter of taiep rats and control animals were found. Nonlinear emissions from the processes and somata of oligodendrocytes in tubulin beta 4a mutant rats were consistently detected, in the shape of elongated structures and cell-like bodies, which were never detected in the controls. This signal represents the second harmonic trademark of the disease. The tissue was also fluorescently labeled and analyzed to corroborate the origin of the nonlinear signal. Besides enabling the description of structural and molecular aspects of H-ABC, our data open the door to the diagnostic use of nonlinear optics in the study of neurodegenerative diseases, with the additional advantage of a label-free approach that preserves tissue morphology and vitality.