MORPHOGENESIS, BIOLOGICAL CYCLES AND PATHOLOGICAL ALTERATIONS IN TISSUES AND ORGANS OF VERTEBRATES

S. Barni - G. Gerzeli - G. Bottiroli - C. Fenoglio - V. Bertone - F. Bernini - R. Vaccarone

The purpose of this study concerns the definition of some morphofunctional characteristics of cellular systems (in vitro) and of tissues and organs (in vivo) related to different kinds of Vertebrates under natural (hibernation) and experimental (stimulation with chemical agents) conditions. The aim of the study, lead with different methodological approaches (fluorescence and electronic microscopy, cytofluorometry and spectrofluorometry, digital image analysis), to underline some functional correlations between nucleus and cytoplasmic and membrane components consequent to the functional state of the cell (differentiation, proliferation, apoptosis). Big part of the researches will concern the deepening of the investigations to liver level in the Amphibians, with the purpose to define the functional interrelations between the different components (hepatocytic, pigmentary and haemopoietic) in the adaptation to the conditions of natural hibernation. Particular space will be devoted to the kinetic activity of the different cellular components of the liver, considering the proliferative aspect (replication of the DNA, caryokinesys and caryodieresys, binuclearisation and polyploidysation) and degenerative apoptotic.
The reliefs related to the haematogenic component of the liver will be correlated with those partly already obtained on other haemopoietic organs and on the peripheral blood.
 
 

HISTOCHEMICAL AND ULTRASTRUCTURAL CHARACTERIZATION OF THE MICROENVIRONMENT OF A SOLID TUMOR AND OF THE EFFECTS INDUCED BY THE TUMOR ON LIVER AND RENAL STRUCTURE AND METABOLISM

I. Freitas - G. Gerzeli - S. Barni - C. Fenoglio - V. Bertone - P. Griffini

It will be continued the study of different microenvironments that are determined within a solid tumor from the heterogeneous nourishment and oxygen supply, from the proximity to the invaded tissues, from the degree of vascularization and interstitial edema etc. to which the malignant cells are adapted through different strategies of survival. The study of such strategies will be conducted with histochemical and ultrastructural techniques. Different enzymatic activities will be shown (dehydrogenase, phosphatase, glucuronidase, phosphorilase, catalase), radicalic species derived by the oxygen and will be identified the prolifering cells through antibodies anti-BrdU. The lipids will be put in evidence with fluorescence microscopy using the probe Nile Red. The mastocytes of connectival and mucosal type will be identified with the method of Alcian Blue-Safranine. Particular attention will be paid to the cells in contact with the edema to document if it determines the formation of oxygen radicals and what the effects are of such radicals on the cells. It will be considered as reference the ultrastructure of the tumor. It will continue besides the study of some effects of the tumor on structure and metabolism of the liver and the kidney, to document how the neoplasia interferes with the normal function of such organs, also in absence of metastasis. The analytical techniques employed will be the same above quoted.

It is foreseen the collaboration with:
- Prof. C.J.F. Van Noorden and Dr. W. Frederiks - Laboratory of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, The Netherlands.
- Dr. H. Gundlach, Carl Zeiss, Jena, Germany.
- Prof. P. J. Stoward, Department of Anatomy and Physiology, University of Dundee, U. K.
- Dr. F. Podo, Laboratorio di Biologia Cellulare, Istituto Superiore di Sanità, Roma.
- Dr. G. F. Baronzio, USSL 34, Legnano.
- Prof. G. Roveta Magrassi, lstituto di Patologia Generale, Università di Pavia.

- Prof. L. Bonandrini, IRCCS Policlinico San Matteo, Pavia.
- Prof. P. Pontiggia, Dipartimento di Termoterapia Oncologica, Clinica "Città di Pavia", Pavia.
 
 

IMMUNOBIOLOGY OF THE TUMORS: MORPHOFUNCTIONAL AND IMMUNOCYTOCHEMICAL STUDY

R. Nano - G. Gerzeli - S - Barni - C. Fenoglio - R. Vaccarone - P. Chiari

It will be continued the in vivo and in vitro study of lymphocytic subpopulations sensitive to interleukin-2 through the use of morphological (also ultrastructural), cytochemical approach and of molecular biology.
Particularly it is intended to deepen the interactions between the tumoral cells of malignant glioma and the cells of the immune system. Using cultures to brief term in vitro, it will be evaluated the proliferation of the tumoral cells and of infiltrating lymphocytes (TIL) in answer to some citokines and to specific drugs. It will be continued the investigation with the electronic microscopy on the mechanism of action and the morphological and functional evaluation of lymphocytes co-cultures with different concentration of IL-2.
The use and the refinement of cytochemical (NOS, 5'-nucleotidase) and immunocitochemical techniques for the study of the cytosckeleton of the cultivated lymphocytes with IL-2 and of the cells of human multiform glioblastoma could furnish useful information on the biology of this tumoral model. Besides on fragments of GBM the expression of intercellular adhesion molecules will be investigated (intercellular adhesion molecules: ICAM) with immunocytochemical methods.

It is foreseen the collaboration with:
- Dr. E. Hair - Dipartimento di Genetica e Microbiologia, Università di Pavia;
- Dr. G. Bottiroli - Centro di Studio per l'Istochimica del C.N.R., Pavia;
- Dr. M. Ceroni - Istituto Neurologico C. Mondino, Università di Pavia.
 
 

CYTOCHEMICAL MARKERS OF CELLULAR FUNCTIONS

P. De Piceis Polver - C. Fenoglio - S. Barni - R. Vaccarone - P. Chiari - G. Gerzeli

A study will be begun related to the collateral effects of the cis-platinum (cytostatic drug) on the muscular fibers of the anterior tibial muscle of rat, taking in consideration especially the mitocondrial alterations, either morphological or functional, and therefore the energetic metabolism. Contemporarily it will be examined, with histochemical and ultrastructural techniques, the effect of the procaine and of the adduct cis-platinum - procaina, to appraise a possible reduction of the collateral effects and therefore to realize an improvement of the therapeutic index.
The toxic effects of the styrene inhalation, chemical compound used in the plastics and resins production, on the liver and on the lung of rat will be also evaluated after administration of styrene with ethanol, to verify a possible interaction of the two substances in determining alterations of the liver and pulmonary parenchyma.
The involvement of the mesentere of Rana esculenta in the phenomenons of the ionic transport asset will be analyzed subsequently with immunocytochemical techniques using an antibody anti-Na+/K+ATPase.

It is foreseen the collaboration with:
- Dr. A. Boicelli - Centro di Studio per l'Istochimica del C.N.R., Pavia;
- Dr. M. Esposito - Istituto Nazionale per la Ricerca sul Cancro, Genova.
 
 

MUSEOLOGICAL RESEARCH

M.V. Gervaso - D. Formenti - C. Violani
in collaboration with C. Garrone, S. Maestri, S. Risi and I. Rognoni.

It is intended to complete the study already undertaken related to the Panizza's manuscripts with the transcription of the block regarding the observations on the heart of the tortoises and on the skin of the african chameleon.
Within the work of exploitation of the historical-scientific museal patrimony of our University, on the occasion of the Conference of Primatology that will be held in Pavia in 1998, it is intended to complete the recovery, the restauration, the catalogatio and the study of some collection of the Primates (skeletal system) of the Comparative Anatomy collection sited to the Visconteo Castle. The study of the Mammals skulls will be completed.
The setup of the collections foresees, besides the study of the historical-scientific aspects, also the planning of exhibition structures and of innovative communication and didactic techniques.

It is foreseen the collaboration with:
- Prof. A. Calligaro - Museo Storico dell'Università di Pavia;
- Dr. T. Rovati - Centro Interdipartimentale Musei, Museo di Storia Naturale, Pavia.