Structural and functional alterations of the cell nucleus in skeletal muscle wasting: the evidence in situ

The histochemical and ultrastructural analysis of the nuclear components involved in RNA transcription and splicing can reveal the occurrence of cellular dysfunctions eventually related to the onset of a pathological phenotype. In recent years, nuclear histochemistry at light and electron microscopy has increasingly been used to investigate the basic mechanisms of skeletal muscle diseases; the in situ study of nuclei of myofibres and satellite cells proved to be crucial for understanding the pathogenesis of skeletal muscle wasting in sarcopenia, myotonic dystrophy and laminopathies

Advances in vascular anatomy and pathophysiology using high resolution and multiparametric sonography

: B-mode and Color Doppler are the first-line imaging modalities in cardiovascular diseases. However, conventional ultrasound (US) provides a lower spatial and temporal resolution (70-100 frames per second) compared to ultrafast technology which acquires several thousand frames per second. Consequently, the multiparametric ultrafast platforms manage new imaging algorithms as high-frequency ultrasound, contrast-enhanced ultrasound, shear wave elastography, vector flow, and local pulse wave imaging. These advances allow better ultrasound performances, more detailed blood flow visualization and vessel walls' characterization, and many future applications for vascular viscoelastic properties evaluation.In this paper, we provide an overview of each new technique's principles and concepts and the real or potential applications of these modalities on the study of the artery and venous anatomy and pathophysiology of the upper limb before and after creating a native or prosthetic arterio-venous fistula. In particular, we focus on high-frequency ultrasound that could predict cannulation readiness and its potential role in the venous valvular status evaluation before vascular access creation; on contrast-enhanced ultrasound that could improve the peri-operative imaging evaluation during US-guided angioplasty; on shear wave elastography and local pulse wave imaging that could evaluate preoperative vessels stiffness and their potential predictive role in vascular access failure; on vector flow imaging that could better characterize the different components of the vascular access complex flow

Current role of ultrasound in hemodialysis access evaluation

: Physical examination (PE) is considered the backbone before vascular access (VA) placement, during maturation period and for follow-up. However, it may be inadequate in identifying suitable vasculature, mainly in comorbid patients, or in detecting complications. This review highlights the advantages of ultrasound imaging to manage VA before placement, during maturation and follow-up. Furthermore, it analyses the future perspectives in evaluating early and late VA complications thank to the availability of multiparametric platforms, point of care of ultrasound, and portable/wireless systems. Technical improvements and low-cost systems should favor the widespread ultrasound-based VA surveillance programs. This significant turning point needs an adequate training of nephrologists and dialysis nurses and the standardization of exams, parameters, and procedures

Basics for performing a high-quality color Doppler sonography of the vascular access

: In the last years, the systematic use of ultrasound mapping of the upper limb vascular network before the arteriovenous fistula (AVF) implantation, access maturation, and clinical management of late complications is widespread and expanding. Therefore, a good knowledge of theoretical outlines, instrumentation, and operative settings is undoubtedly required for a thorough examination. In this review, the essential Doppler parameters, B-Mode setting, and Doppler applications are considered. Basic concepts on the Doppler shift equation, angle correction, settings on pulse repetition frequency, operative Doppler frequency, gain are reported to ensure adequate and correct sampling of blood flow velocity. A brief analysis of the Doppler inherent artefacts (as random noise, blooming, aliasing, and motion artefacts) and the adjustment setting to minimize or eliminate the confounding artefacts are also considered. Doppler aliasing occurs when the pulse repetition frequency is set too low. This artefact is particularly frequent in vascular access sampling due to the high velocities range registered in the fistula's different segments. Aliasing should be recognized because its correction is crucial to analyse the Doppler signals correctly. Recent advances in instrumentation are also considered about a potential purchase of a portable ultrasound machine or a top-of-line, high-end, or mid-range ultrasound system. Last, the pulse wave Doppler setting for vascular access B-Mode and Doppler assessment is summarized

Routinely frozen biopsies of human skeletal muscle are suitable for morphological and immunocytochemical analyses at transmission electron microscopy

The aim of the present investigation was to evaluate whether routinely frozen biopsies of human skeletal muscle may be suitable for morphological and immunocytochemical analyses at transmission electron microscopy. The fixation/embedding protocols we successfully used for decades to process fresh mammalian tissues have been applied to frozen muscle biopsies stored for one to four years in liquid nitrogen. After 2.5% glutaraldehyde -2% paraformaldehyde - 1% OsO4 fixation and embedding in epoxy resin, the ultrastructural morphology of myofibres and satellite cells as well as of their organelles and inclusions proved to be well preserved. As expected, after 4% paraformaldehyde - 0.5% glutaraldehyde fixation and embedding in LR White resin, the morphology of membrane-bounded organelles was relatively poor, although myofibrillar and sarcomeric organization was still recognizable. On the contrary, the myonuclei were excellently preserved and, after conventional staining with uranyl acetate, showed an EDTA-like effect, i.e. the bleaching of condensed chromatin, which allows the visualization of RNP-containing structures. These samples proved to be suitable for immunocytochemical analyses of both cytoskeletal and nuclear components, whereas the poor mitochondrial preservation makes unreliable any in situ investigation on these organelles