Inflammatory effects on human lung epithelial cells after exposure to diesel exhaust micron sub particles (PM1.0) and pollen allergens.

Asthma is currently defined as a chronic inflammatory disease of the airway. Several evidence indicate that vehicle emissions in cities is correlated with the allergic respiratory diseases. In the present study, we evaluated in the A549 cells the production and release of IL-4, IL-5 and IL-13 after treatment with sub-micron PM(1.0) particles (PM(1.0)), Parietaria officinalis (ALL), and PM(1.0) + ALL together. Our data demonstrated that PM(1.0) + ALL together exhibited the greatest capacity to induce A549 cells to enhance the expression of IL-4 and IL-5 compared with the only PM(1.0) or ALL treatment. Interestingly, IL-13 that is necessary for allergen-induced airway hyper responsiveness, is increased in cells treated with PM(1.0) + ALL together, but is higher expressed when the cells are treated only with the allergen. Our data support the hypothesis that the urban environment damage the acinar lung units and activates cells of the immune system.

Effects of diesel exhaust particles on human lung epithelial cells: an in vitro study.

Atmospheric particulate matter (PM), an ingredient of urban pollution matter, is a mixture of solid and liquid particles differing in origin, dimension and composition. There is big concern about inhaled PM in urban areas, especially due to its adverse effects on the respiratory system. Diesel exhaust particulate (DEP), which constitutes the major part of PM, is characterized by a carbonic mixture composed of approximately 18,000 different high-molecular-weight organic compounds. Diesel engines release 10 times the amount of NO(2) aldehydes and breathable PM compared to unleaded gasoline engines and more than 100 times that produced by catalysed gasoline engines; these data gain great significance when taken into account the fact that diesel-powered vehicles are becoming more and more popular. DEP polyaromatic hydrocarbons (PAH), once deposited on airways mucous surfaces easily pass through epithelial cells (ECs) membranes, bind themselves to cytosolic receptors and then affect cell growth and differentiation. Human lung epithelial cells and macrophages engulf DEP, this resulting in increased proinflammatory cytokines release (IL-6, IL-8 and GM-CSF). We investigated the biological effects of DEP-PM on the human lung EC line A549. Light microscopy analysis suggested the presence of cell wall alterations, and provided evidence of PM internalization and cytoplasmic vacuolization. Following PM stimulation, nuclei also were seen undergo clear gross morphological modifications. Immunocytochemistry was used to detect intracytoplasmic IL-6 and IL-8 expression.

Hyaluronic acid based materials for intestine tissue engineering: a morphological and biochemical study of cell-material interaction.

A wide number of gastro-intestinal disorders are associated with structural alterations of this district leading to an impaired gastrointestinal function. The study of cell material interactions represents one of the major issues for the development of tissue engineering purposes. Benzyl esters of hyaluronic acid are promising materials because they exhibit good tissue compatibility and are available in various configurations. In this work they have been studied for the possible application of intestinal cell growth and functioning. The preliminary investigation on the morphologic and biochemistry data obtained by monitoring the growth and differentiation of intestinal epithelial cells on two hyaluronic acid benzyl esters is reported. Two types of materials structures were studied: a three dimensional matrix and a macroporous flat sheet membrane. Caco-2 cell line was used: these cells undergo spontaneous enterocytic differentiation after several days in culture. The differentiation status of these cells grown on different materials was used as a parameter of biocompatibility and cell functioning. The status of cell growth and differentiation was monitored by studying cell morphology using scanning electron microscopy. The results obtained were confirmed by biochemical determinations. Although both the configurations of the two polymers exhibited good compatibility with respect to intestinal cells, only the flat sheet membrane proved to induce cell differentiation, leading us to the conclusion that it is a promising substrate for the proposed application.

Potential role for high and low molecular weight tissue transglutaminases in transforming mammalian cell properties.

Tissue transglutaminase (tTGase, tTG) is known as being implicated in the intracellular cross-linking of proteins occurring in a growing series of physiological conditions including--just to mention the most relevant ones--programmed cell death (apoptosis), cell adhesion, growth, spreading and differentiation, tumor growth, metastasis, cell adhesion, proliferation, differentiation, extra cellular matrix (ECM) stabilization. In the current work we investigated tTG activity and expression of "normal" and potential transformed cytosolic tTG antigens in mammalian cells. Most cell lines studied showed low tTG activity, which in all cases could be enhanced considerably by treating cell cytosol homogenates with trypsin. The results suggested the existence -in transformed cells- of inactive types of tTGase. We purified cytosolic tTG antigens from these cells utilizing a GTP-agarose resin, and we can therefore conclude that "normal" molecular weight (mw) tTG antigens, but also high molecular weight (hmw) and low molecular weight (lmw) tTG antigens from transformed cells, retain GTP-binding ability. The initial results from our study also allowed us to hypothesize that transformed hmw- and lmw- tTG antigens should not be considered as the result of post-translational modifications of normal mw, cytosolic tTG. The potentially low or absent transamidating functionality of cytosolic tTG species in transformed mammalian cells could be responsible for decreased or even abolished programmed cell death, whereas the unaffected GTP-binding functionality of such proteins in these cells might lead to increased signal transduction and possibly proliferation.

Bartolomeo Eustachio: A pioneer in morphological studies of the kidney.

The morphological structure of the kidney only started being seriously investigated and truly understood in the 16th century, thanks mainly to Bartolomeo Eustachio. In his De Renibus, in fact, he painstakingly described the size, consistency, location and variations of the kidney, to which modern understanding can add very little. In describing the renal parenchyma he stated that it is made up of an external and an internal substance, and recognized the central role of the renal arteries in the excretory function of the kidneys. He made observations regarding the presence of extremely fine arteries that filter urine, the nature and function of the renal tubules and the columns of external substance that protrude between the papillae. There can be no doubt that Eustachio's remarkable achievements made him a pioneer in morphological studies of the kidney.

Marcello Malpighi (1628-1694).

The current paper is intended as a short but precise illustration of Marcello Malpighi's cultural life, a small tribute we deserve to a genius of such sharp insight. He made many discoveries in the fields of the macro- and microanatomy of the brain, nerves, liver, kidneys, spleen, tegument, lymph nodes, reproductive system and other organs, but rather than pursuing these in any detail, we have tried to profile Marcello Malpighi's scientific life in the cultural context of his time, while also providing some information on the most fascinating phases of his research on renal structure and function.

The structure of lungs and kidneys in the work of Malpighi.

This work documents the fundamental contribution to the knowledge of lungs and kidneys made by Marcello Malpighi. For the first time, in "De Pulmonibus", he described the pulmonary alveoli and the network of capillary perialveolar blood vessels, establishing definitively the existence of a communication between arteries and veins. Furthermore, he demonstrated that air does not enter the blood vessels, but that it makes contact solely with the thin walls of the delicate perialveolar vessels. Malpighi in "De Renibus" exactly described the microscopic anatomy of the cortex of the kidney, composed of renal glomerules. Then he explained the function of these minute structures where the urine is separated from the blood passing into the renal tubules.

Anatomic variations of the deep palmar arteries in man.

The deep palmar circulation is constituted by the deep palmar arch. In most cases this is a complete arch formed by the radial artery and its continuation to a deep branch of the ulnar artery. In a few cases, the deep palmar circulation is formed only by the radial or the ulnar artery. Only rarely is there a complete absence of the deep palmar arch. A series of 60 vascular casts was examined in order to identify the primary variants of the deep palmar arterial supply. Four anatomic patterns were identified: (1) radioulnar (66.67%); (2) radial-anastomotic (21.67%); (3) radial (8.33%), and (4) ulnar (3.33%). Two distinct types of the radioulnar variant were observed, a proximal and a distal one, named according to the origin of the deep palmar branch.

Retroviral vector-mediated gene transfer into human primary myogenic cells leads to expression in muscle fibers in vivo.

Primary human myogenic cells isolated from fetal and adult muscle were infected with a high-titer, Moloney murine leukemia virus (MoMLV)-derived retroviral vector expressing a bacterial beta-galactosidase (beta-gal) gene under long terminal repeat (LTR) control. Gene transfer efficiency averaged 50% in both fetal myoblasts and adult satellite cells, as revealed by beta-gal staining. The reporter gene was stably integrated, faithfully inherited, and expressed at significant levels in myogenic cells for at least 10 generations under clonal growth conditions, and throughout the culture life span upon differentiation into myotubes. Comparable gene transfer efficiency was obtained in myogenic cells from muscle biopsies of patients affected by a number of genetic or acquired myopathies, including Duchenne muscular dystrophy. Transduced normal human satellite cells were injected into regenerating muscle of immunodeficient mice, where they formed new muscle fibers in which the product of the reporter gene was detectable for 2 months after injection. These results show that retroviral vectors can be used to transfer foreign genes with high efficiency into normal or abnormal primary human myogenic cells, leading to stable expression into mature muscle. Satellite cells engineered in this way might represent an effective tool for gene therapy of muscular dystrophies as well as for systemic delivery of recombinant gene products for correction of inherited and acquired disorders. The human-mouse model described here will allow in vivo testing of such gene therapy approaches.

A contribution to the history of renal structure knowledge (from Galen to Malpighi).

This work documents the progressive gain in knowledge on renal anatomy acquired by anatomists from Galen to Malpighi. Galen, with albeit his rather imaginative explanations, was the first anatomist to recognize the urine producing function of the kidney. His influence was felt up to the Middle Ages; his followers imagined the presence of two cavities within the kidney that were separated by a porous membrane that they called the "colatorium" which was capable of filtering the urine from the blood. It was only later that Berengario da Carpi, divorcing himself from Galenic dogmatism, finally dismissed the presence of the colatorium. He described the renal papillae and gave the first elementary model of renal vascularization. Further important progress was due to the studies of Falloppius and Eustachius who brought contemporary understanding of renal structure to the limit of what could be seen with the unaided eye. They distinguished the difference between the unilobar canine kidney and the human multilobar organ, they also described the minor and major calyces and, furthermore, guessed at the canalicular structure of its parenchyma. Highmore was then responsible for the description of the archiform vessels which he hypothesized as an arterio-venous anastomotic net between the renal cortex and medulla. With the invention of the microscope, new doors opened for the study of renal structure. Bellini proved the canalicular organization of the parenchyma and, moreover, described the interlobular vessels. Malpighi then described the glomerulus and its relation to the intrarenal excretory ducts. The basis had now been laid for the beginning of modern nephrology.

Paracrine stimulation of senescent satellite cell proliferation by factors released by muscle or myotubes from young mice.

The proliferative potential of satellite cells undergoes a dramatic decrease in the early postnatal period and a more modest but continuous decrease throughout the life span of the animal. To address the problem of the mechanism regulating this phenomenon and to understand whether it is causally linked to senile muscle atrophy, we studied the response of aged satellite cells to serum and to different growth factors. The data reported indicate a generalised reduction in the response to all mitogens tested, which could not be compensated for by increased concentrations of serum or growth factors. On the other hand, conditioned medium of differentiated myotubes from young mice exhibited a strong mitogenic action on aged satellite cells, while conditioned media of myotubes from old mice or from a variety of non-muscle cells were ineffective. Furthermore, saline extracts from muscle of young mice are also able to exert this mitogenic action. Saline extracts of muscle from old mice were poorly mitogenic for satellite cells from young mice, and not at all for satellite cells from old mice. These data indicate that paracrine interactions operate inside the muscle tissue and are probably required for the normal replicative behaviour of satellite cells. The failure of such interactions may be among the causes leading to age-related muscle hypotrophy.

The effects of AZT and DDI on pre- and postimplantation mammalian embryos: an in vivo and in vitro study.

This study reports the effects of the nucleoside analogs dideoxyinosine (DDI) and 3'-azido-3'-deoxythymidine (AZT) on mammalian embryonic development. When administered to pregnant mice (at concentrations ranging from 10 to 300 mg/kg/day), through all or part of gestation, AZT and DDI did not result in any visible effect on mouse embryos nor did they cause any obvious malformation or defect at birth or during postnatal growth. Similarly, when embryonic or fetal mouse or human cells (from brain, limb buds, or different organ rudiments) were exposed to AZT or DDI in vitro, cytotoxicity was observed only in the mM range, with AZT showing slightly higher cytotoxicity and brain cells appearing slightly more sensitive to both nucleosides. However, even in cultures treated with very high concentrations of AZT or DDI, the reduction in the number of terminally differentiated skeletal myotubes, cardiocytes, neurons, and chondrocytes was similar to the reduction in the total number of cells, indicating that AZT and DDI did not selectively inhibit differentiation of any of the above-mentioned cell types. Finally, preimplantation mouse embryos (at the 2-cell or 4-cell stage), treated in vitro with micromolar concentrations of AZT, were arrested at the 4-cell stage. DDI or other nucleoside analogs tested did not have this effect.

An atypic pattern of the azygos venous system in man.

A rare variation of the azygos system, found in an 84 year old man, is described. A "double-columned" azygos system runs in front of the spine, and the azygos major crosses the superior lobe of the right lung and enters the right subclavian vein.

A new approach to the vascular-nervous correlations of the superior mediastinum in man.

The relationship between the superior mediastinal organs has been investigated in the adult and the newborn. It was found that this region could be represented by a geometrical schematization based on the correlations that exist among the vascular and nervous elements that lie on the right mediastinal wall. A vascular triangle, a vascular-nervous triangle and a retro-vagal square are described. The size of these forms and their relations with the organs they enclose varies depending on the build of the subject being examined in this area, particularly from a teaching point of view.