Clathrin-dependent endocytosis of membrane-bound RANKL in differentiated osteoclasts

Bone is continuously repaired and remodelled through well-coordinated activity of osteoblasts that form new bone and osteoclasts, which resorb it. Osteoblasts synthesize and secrete two key molecules that are important for osteoclast differentiation, namely the ligand for the receptor of activator of nuclear factor κB (RANKL) and its decoy receptor osteoprotegerin (OPG). Active membrane transport is a typical feature of the resorbing osteoclast during bone resorption. Normally, one resorption cycle takes several hours as observed by monitoring actin ring formation and consequent disappearance in vitro. During these cyclic changes, the cytoskeleton undergoes remarkable dynamic rearrangement. Active cells show a continuous process of exocytosis that plays an essential role in transport of membrane components, soluble molecules and receptor-mediated ligands thus allowing them to communicate with the environment. The processes that govern intracellular transport and trafficking in mature osteoclasts are poorly known. The principal methodological problem that have made these studies difficult is a physiological culture of osteoclasts that permit observing the vesicle apparatus in conditions similar to the in vivo conditions. In the present study we have used a number of morphological approaches to characterize the composition, formation and the endocytic and biosynthetic pathways that play roles in dynamics of differentiation of mature bone resorbing cells using a tri-dimensional system of physiologic coculture

Interferon-mediated intracellular signalling Modulation of different phospholipase activities in Burkitt lymphoma cells

AbstractThe effect of interferon-α on Daudi lymphoma cells either sensitive or resistant to the action of this cytokine has been analysed in terms of phospholipase C (PLC) and D (PLD) activities. Results have shown a combined modulation of PIP2-specific phospholipase C and phospholipase D. In particular, a decreased activity of PIP2-specific PLC has been found, concomitant to a PLD-mediated phosphatidylcholine hydrolysis, suggesting that the intracellular signaling activated by interferon in Daudi cells involves a phospholipase D/phosphohydrolase pathway

Lamellar and «club-shaped» corpuscular nerve endings in human gingival mucosa. A light and electron microscopic study

A study on the presence of corpuscular nerve endings in human gingival mucosa was performed using both light and transmission electron microscopic (TEM) techniques. Both round and oval lamellar corpuscles were detected by light microscopy. They were located either subepithelially, close to the basement membrane, or within the papillae, deeply invaginated into the overlying epithelium. TEM techniques showed convoluted structures with unmyelinated fibre arborizations leading to an afferent fibre supported by the so called lamellar cells. The presence of blood vessels, collagenous fibrils, desmosome-like junctions, cytoplasmic organelles, as well as the similarity with some previously described mechanoreceptors, suggested the role of such corpuscular nerve endings in transmitting a nervous impulse induced by mechanical stimulation. Other simpler structures were also observed and named «club-shaped» corpuscles: they could support the more complex ones in responding to the strengths and the movements directly influencing the gingival mucosa.La présence de terminaisons nerveuses corpusculaires dans la muqueuse gingivale humaine a été observée tant en microscopie optique qu’en microscopie électronique à transmission. En microscopie optique on a remarqué des corpuscules lamellaires ronds et ovalaires, qui étaient localisés tant au dessous de l’épithélium, tout près de la membrane basale, qu’au dedans des papilles, profondément insérés dans l’épithélium.En microscopie électronique on a observé des structures convolutées pourvues d’arborisations de fibres nerveuses sans myéline qui vont se réunir dans une fibre afférente supportée par des cellules dites lamellaires. La présence de vaisseaux, de fibrilles collagènes, de jonctions telles que desmoses, d’inclusions cytoplasmiques autant que la ressemblance avec quelques mécanorécepteurs décrits en littérature, suggérait un rôle de ces terminaisons nerveuses corpusculaires en envoyant un impulse nerveux induit par une stimulation mécanique. On a aussi observé des corpuscules plus simples appelés «club-shaped» qui pourraient supporter les plus complexes dans la réponse aux forces et aux mouvements qui influencent directement la muqueuse gingivale

CT Guided fine needle aspiration biopsy of pulmonary lesions under 15 mm of diameter: results on 68 consecutive patients

BACKGROUND Most reports on lung fine needle aspiration cytology (FNAC) demonstrate that the diagnostic accuracy tends to decrease with the size of the lesions, and that the frequency of complications increases as the size of lesions decreases. PURPOSE The aim of this prospective study was to describe the accuracy and incidence of complications related to FNAC of solitary pulmonary nodules (SPNs) of 15 mm or less in diameter. Moreover, we evaluated how this procedure during the initial evaluation of patients with SPN can reduce the number of unnecessary surgery. MATERIAL AND METHODS From January 2012 to December 2014, 225 patients with an SPN between 7-15 mm in diameter were referred to our Institution. Patients with risk factors such as ASA 3, FEV1

Excitability and Synaptic Alterations in the Cerebellum of APP/PS1 Mice

In Alzheimer's disease (AD), the severity of cognitive symptoms is better correlated with the levels of soluble amyloid-beta (Aβ) rather than with the deposition of fibrillar Aβ in amyloid plaques. In APP/PS1 mice, a murine model of AD, at 8 months of age the cerebellum is devoid of fibrillar Aβ, but dosage of soluble Aβ1–42, the form which is more prone to aggregation, showed higher levels in this structure than in the forebrain. Aim of this study was to investigate the alterations of intrinsic membrane properties and of synaptic inputs in Purkinje cells (PCs) of the cerebellum, where only soluble Aβ is present. PCs were recorded by whole-cell patch-clamp in cerebellar slices from wild-type and APP/PS1 mice. In APP/PS1 PCs, evoked action potential discharge showed enhanced frequency adaptation and larger afterhyperpolarizations, indicating a reduction of the intrinsic membrane excitability. In the miniature GABAergic postsynaptic currents, the largest events were absent in APP/PS1 mice and the interspike intervals distribution was shifted to the left, but the mean amplitude and frequency were normal. The ryanodine-sensitive multivescicular release was not altered and the postsynaptic responsiveness to a GABAA agonist was intact. Climbing fiber postsynaptic currents were normal but their short-term plasticity was reduced in a time window of 100–800 ms. Parallel fiber postsynaptic currents and their short-term plasticity were normal. These results indicate that, in the cerebellar cortex, chronically elevated levels of soluble Aβ1–42 are associated with alterations of the intrinsic excitability of PCs and with alterations of the release of GABA from interneurons and of glutamate from climbing fibers, while the release of glutamate from parallel fibers and all postsynaptic mechanisms are preserved. Thus, soluble Aβ1–42 causes, in PCs, multiple functional alterations, including an impairment of intrinsic membrane properties and synapse-specific deficits, with differential consequences even in different subtypes of glutamatergic synapses

Protein kinase C and cardiac dysfunction: a review

Heart failure (HF) is a physiological state in which cardiac output is insufficient to meet the needs of the body. It is a clinical syndrome characterized by impaired ability of the left ventricle to either fill or eject blood efficiently. HF is a disease of multiple aetiologies leading to progressive cardiac dysfunction and it is the leading cause of deaths in both developed and developing countries. HF is responsible for about 73,000 deaths in the UK each year. In the USA, HF affects 5.8 million people and 550,000 new cases are diagnosed annually. Cardiac remodelling (CD), which plays an important role in pathogenesis of HF, is viewed as stress response to an index event such as myocardial ischaemia or imposition of mechanical load leading to a series of structural and functional changes in the viable myocardium. Protein kinase C (PKC) isozymes are a family of serine/threonine kinases. PKC is a central enzyme in the regulation of growth, hypertrophy, and mediators of signal transduction pathways. In response to circulating hormones, activation of PKC triggers a multitude of intracellular events influencing multiple physiological processes in the heart, including heart rate, contraction, and relaxation. Recent research implicates PKC activation in the pathophysiology of a number of cardiovascular disease states. Few reports are available that examine PKC in normal and diseased human hearts. This review describes the structure, functions, and distribution of PKCs in the healthy and diseased heart with emphasis on the human heart and, also importantly, their regulation in heart failure