High-dimensional immunotyping of tumors grown in obese and non-obese mice.

Obesity is a disease characterized by chronic low-grade systemic inflammation and has been causally linked to the development of 13 cancer types. Several studies have been undertaken to determine whether tumors evolving in obese environments adapt differential interactions with immune cells and whether this can be connected to disease outcome. Most of these studies have been limited to single-cell lines and tumor models and analysis of limited immune cell populations. Given the multicellular complexity of the immune system and its dysregulation in obesity, we applied high-dimensional suspension mass cytometry to investigate how obesity affects tumor immunity. We used a 36-marker immune-focused mass cytometry panel to interrogate the immune landscape of orthotopic syngeneic mouse models of pancreatic and breast cancer. Unanchored batch correction was implemented to enable simultaneous analysis of tumor cohorts to uncover the immunotypes of each cancer model and reveal remarkably model-specific immune regulation. In the E0771 breast cancer model, we demonstrate an important link to obesity with an increase in two T-cell-suppressive cell types and a decrease in CD8 T cells.

Ion transport proteins and aquaporin water channels in the kidney of amphibians from different habitats.

Amphibians are known to spend part of their life on land and return to water to reproduce. However, some urodeles spend their entire life in water, while others succeed in completely avoiding water even during reproduction. Osmoregulatory mechanisms must therefore be different in the diverse environmental conditions of their respective life histories. The architecture of the kidney is similar in all amphibians; as a consequence the ion-water equilibrium must be regulated in the different environmental conditions. We investigated the immunolocalisation of Na(+)/K(+)/Cl(-) cotransport proteins, sodium pump and water-channel proteins (aquaporins) in aquatic Amphiuma means means, Rana dalmatina, a species that returns to water to reproduce, and Speleomantes genei, a completely terrestrial species. The investigation was carried out with immunohistochemical methods using antibodies to Na(+)/K(+)/Cl(-) cotransport protein NKCC1 T4, Na(+)/K(+)ATPase alpha-subunit, water-channel aquaporin 3 and the inner mitochondrial membrane (AMA). Cotransport proteins and sodium pump, involved in ion reabsorption, are widely distributed in A. means and R. dalmatina and confined to the distal segment in S. genei; conversely water channels, involved in water reabsorption, are limited to the collecting duct in A. means and R. dalmatina and distributed in the proximal and collecting ducts in S. genei.

Soft tissue sarcomas: research of prognostic factors and therapeutics scheme.

The Soft Tissue Sarcomas shows consisting mortality that grazes 50%. Commonly the histologic type, the largeness of the tumor and the degree of malignity, are considered prognostic factors influencing the survival of patients affected from such tumor. The aim of our study is to characterize the new prognostic factors influencing the local recidives and metastatic dissemination. We have analysed 17 possible clinical and histologic prognostic factors in 80 patients affected of soft tissue sarcomas. The results have dimonstrated that: the compartimentality and the cellular differentiation seem to be more influent to the insurgence of local recidives and metastasis; while the low malignity degree, the wide surgical margins and the histopatological characters of benignity can influense positively the survival.

Microgravity-induced apoptosis in cultured glial cells.

Apoptosis is a form of naturally occurring cell death that plays fundamental roles during embryonic developement. In adults, it neatly disposes of cells damaged by injuries provoked by external causes such as UV radiation, ionisation and heat shock. Alteration of the gravity vector may be one of the external apoptosis inducers. Neurophysiological impairment signs were seen during space flights in astronauts, but very few studies were carried out on the nervous system and none at the cellular level. In this study, we submitted cultured C6 glioma cells to microgravity (0xg) of varying duration, obtained by clinorotation in a Fokker three-dimensional clinostat for 15 min, 30 min, 1h, 20h or 32h. After 30 min at 0xg, numerous nuclei underwent the classical morphological alterations (chromatin condensation, nuclear fragmentation, apoptotic bodies) that lead to the programmed cell death. After 30 min at 0xg, immunostaining for the enzyme caspase-7 was present in the cytoplasm of many cells concurrently with DNA fragmentation identified by the TUNEL method. At 32h, the number of apoptotic nuclei was much reduced indicating the ability of glial cells to adapt to altered gravity.

Diffuse hyperplastic oncocytosis of the parotid gland.

Oncocytic tumours rarely affect the major salivary glands, accounting for less than 1% of all salivary-gland tumours. The World Health Organisation classification groups these tumours into three principal types: diffuse oncocytosis, focal adenomatous oncocytic hyperplasia and oncocytoma. Diffuse hyperplastic oncocytosis is the rarest lesion: only six cases have been previously reported in the literature. This condition of putative hyperplastic pathogenesis follows a benign course, whereas oncocytomas may recur after excision. No metastatic dissemination or recurrence of diffuse hyperplastic oncocytosis has been reported. We present and discuss a new case of diffuse hyperplastic oncocytosis of the parotid gland.

Microgravity-induced programmed cell death in astrocytes.

Cultured astrocytes were submitted to simulated microgravity using a Fokker clinostat under continuous rotation (60 rpm) for 15', 30', 1h, 20h and 32h. Samples processing included (i) nuclear stainings using Propidium Iodide and 4,6-diamidino-2-phenilindole, dihydro chloride, (ii) immunohistochemical identification of Caspase-7, (iii) identification of DNA fragmentation using the terminal dUTP nick end labelling and (iv) Scanning Electron Microscope analysis. After 30' at simulated microgravity the glial cells showed morphological evidence of apoptosis: cell shrinkage, chromatin condensation, nuclear blebs and fragmentation. The enzyme caspase-7 was present and DNA fragmentation was evident. After 32h the density of the cell population was much lower than that observed in controls.

Mitochondria-rich cells in gills and skin of an African lungfish, Protopterus annectens.

We used scanning electron microscopy, the vital dye DASPEI and an antibody to the inner mitochondrial membrane to study the presence and localisation of mitochondria-rich cells in the gills and skin (opercular, dorsal and ventral) of the lungfish Protopterus annectens in its free-swimming conditions and at the beginning of aestivation. In the free-swimming period, the gills were short and thick and the pavement cells were extremely large (30-40 microns). The mitochondria-rich cells, which were distributed in the secondary and primary epithelium, occurred as two morphologically different types, i.e. elongated and oval, similar to the alpha and beta chloride cells of fresh water teleosts. In the skin, only one type of mitochondria-rich cells was found, resembling the alpha chloride cells. All the mitochondria-rich cells distributed in the gills and skin were labelled with anti Ca(2+)-ATPase serum indicating the possible uptake of Ca2+ at freshwater chloride cell level. At the start of aestivation, the skin and gills were covered by a thick layer of mucus and the epithelium of the gills was reduced. The mitochondria-rich cells were almost completely covered by the pavement cells.

Simulated microgravity induces alteration in the central nervous system.

To investigate whether the signs of neurophysiological impairment observed in flight may be traced back to cytomorphology, we undertook a ground-based study focusing upon the architecture of cultured glial cells under simulated microgravity obtained by three-dimensional clinorotation.

The effect of twisting on microanastomotic patency of arteries and veins in a rat model.

The authors examined the effect of twisting on the patency of microvascular anastomoses 3 days after surgery. A total of 69 male Wistar rats were divided randomly into four groups. The femoral arteries and veins were dissected for a standard distance. A total of 69 microarteriorrhaphies and 68 microvenorrhaphies were performed at 0 deg and with twist of the vessel ends of 90, 180, and 270 deg. Three-day patency rates for arterial microanastomoses were 100% with a 0-deg twist, 80.9% with a 90-deg twist, 68.4% with a 180-deg twist, and 64.2% with a 270-deg twist. Three-day patency rates for venous microanastomoses were 100% with a 0-deg twist, 85% with a 90-deg twist, 28.5% with a 180-deg twist, and 25% with a 270-deg twist (p = 0.047 for arteries, p = 0.001 for veins). These data are statistically significant. Moreover, assuming the risk of thrombosis to be 1 for microanastomosis without twisting, the odds ratio for the risk of vessel thrombosis for 270-deg twisting (the maximal examined degree of arterial and venous twist in the current study) is 10.08 for arterial anastomosis and 226.85 for venous anastomosis.

Somatostatin in the ovary of an African lungfish (Protopterus annectens): an in situ hybridisation, immunohistochemical, and autoradiographical study.

In mammals, somatostatin seems to be involved in the control of ovarian steroidogenesis. There have been no studies on the presence or actions of somatostatin in the ovary of nonmammalian vertebrates. The localisation of somatostatin-14 was examined immunohistochemically using the antibody to somatostatin-14 in the ovary of the African lungfish Protopterus annectens. Immunoreactivity was present in the granulosa cells of mature ovarian follicle examined by light microscopy. Using an oligonucleotide probe complementary to mRNA for somatostatin-14 and labelled at the 3'-end with alpha-35S, in situ hybridisation demonstrated somatostatin-14 mRNA distributed in cells showing the same localisation as that of the immunoreactive cells. Binding sites for SST-14 were identified with autoradiography using [125I]somatostatin-14. Binding sites were localised on granulosa and theca cells. Somatostatin-14 may be thus synthesised in the lungfish ovary.

Proopiomelanocortin (POMC) mRNA and POMC-derived peptides immunolocalization in the skin of Protopterus annectens, an African lungfish.

Antisera against adrenocorticotropic hormone (ACTH), alpha-melanocyte stimulating hormone (alpha-MSH) and beta-endorphin were used to localize, by immunohistochemistry, proopiomelanocortin (POMC)-derived peptides in the skin excised from different regions of the African lungfish Protopterus annectens. Immunoreactivity was observed in the epidermis mainly in the germinal layer. Using human POMC cDNA as hybridization probe, POMC-like mRNA was identified in situ in epidermal cells. The demonstration in the same cells of POMC mRNA and POMC-related peptides immunoreactivity indicates a local production of opiate hormones.

Analysis of infiltrating epitheliomas of the nose examined from 1986 to 1995.

OBJECTIVE: Epitheliomas of the nose are frequently highly aggressive lesions with wide cutaneous-subcutaneous growth or an infiltrative growth pattern, especially when located at the columellar base, the alar base, and toward the inner chantal region. The aim of this study was to review this lesion with respect to location, recurrence, and treatment. METHOD: The authors present a retrospective study of 193 infiltrating epitheliomas of the nose examined between 1986 and 1995, including primary and recurrent cases. The authors term "clinically infiltrating epithelioma" any epithelioma whose growth implies the infiltration of the osteocartilaginous framework. RESULTS: The analysis of this case series, based on the histotype and the site of the epitheliomas, confirms that most of the primary histologically infiltrating basal cell carcinomas (BCCs) and of the recurrent BCCs are located at the alar base. The analysis of recurrences shows an aggressive local behaviour (multiple recurrences in the same site, change of the histologic pattern toward a more malignant one, etc.) suggesting that many of them were submitted to an insufficient physical or surgical treatment in the past. CONCLUSIONS: A radical surgical approach at the first signs of the primary lesion is mandatory in cases of nasal epithelioma.

Expression of pro-opiomelanocortin (POMC) in the cerebral ganglion and ovary of a protochordate.

The distribution of neurones expressing POMC mRNA in the cerebral ganglion of the protochordate ascidian, Styela plicata, was investigated using a non-radioactive in situ hybridization technique. Nerve cell bodies of mono and bipolar types expressing POMC mRNA, were observed mainly in the outer layer of the ganglion. Discrete groups of neurones containing POMC mRNA were also localized in the inner portion of the ganglion, and few small monopolar perykaria expressing POMC mRNA were visible at the emergence of the main nerve trunks. POMC mRNA labeling was also found at level of the cytoplasm of previtellogenic and vitellogenic oocytes, and of follicular cells. Our results demonstrate the expression of one or more genes in the cerebral ganglion and ovary, that may be similar to one or more regions of the mammalian POMC gene. Therefore POMC-related molecules seem to be involved in neuromodulatory pathways and regulatory mechanisms of the oogenesis of ascidians.

Somatostatin in lungfish kidney: an immunohistochemical, autoradiographical and in situ hybridisation study.

The localisation of somatostatin-14 (SST-14) was examined immunohistochemically using the antibody Ab-SST-14 in the kidney of the African lungfish Protopterus annectens. Immunoreactive cells were present in the proximal tubules. In situ hybridisation, using an oligonucleotide probe complementary to mRNA for SST-14 and labeled at the 3'-end with alpha-35S, showed SST-14 mRNA distributed in cells with the same localisation as seen for SST-14 immunoreactive cells. Binding sites for SST-14 were identified with autoradiography using 125I SST-14. Binding sites were concentrated on cells of the proximal tubules. It is suggested that SST-14 may be synthesised in the lungfish mesonephros.

Malignant tumors of the orbital region. Analysis of cases examined from 1986 to 1995.

OBJECTIVE: To present a retrospective study of 154 orbital tumors examined from 1986 to 1995. METHODS: We analyzed the clinical behavior and the degree of local malignity also in relationship with the particular anatomical characteristics of this site. RESULTS: In this are, it is not rare finding tumors with a scarce cutaneous extension but a deep involvement including the bone, or of recurrences after a previous insufficient treatment with an aggressive clinical course. CONCLUSION: The principles for the surgical approach are: exeresis as radical as possible, one-stage operation for tumoral resection and reconstruction, bringing of trophic tissue able to sustain any complementary treatment, easy and fast execution, and short clinical course.

Localization of interleukin-1 beta mRNA in the cerebral ganglion of the protochordate, Styela plicata.

There is evidence that interleukin 1 beta (IL-1 beta) plays an important role in several biological functions in mammals where it is synthesized by cells of haematological, dermal and neural origin. Moreover, production of cytokine-like molecules has been demonstrated in some blood cells of non-mammalian vertebrates and invertebrates in which also nerve cells are demonstrated to be IL-1 beta immunoreactive. The purpose of the present study is to demonstrate the IL-1 beta mRNA expression in nerve cells of the ascidian Styela plicata by use of non-isotopic in situ hybridization technique. The expression of IL-1 beta messenger was demonstrated in monopolar neurons in the cortical layer of the cerebral ganglion. The neuronal expression of cytokine-like molecules in tunicates suggests that IL-1 beta is an ancestral and functionally conserved molecule, and that a neuroimmune axis appeared early during the metazoan phylogeny.

Pro-opiomelanocortin (POMC) expression and immunolocalization of POMC-related peptides in the ovary of Protopterus annectens, an African lungfish.

Antisera against adrenocorticotropic hormone (ACTH), alpha-melanocyte-stimulating hormone (alpha MSH) and beta-endorphin were used to localize pro-opiomelanocortin (POMC)-derived peptides in the ovary of the African lungfish Protopterus annectens by immunohistochemistry. Immunoreactivity was observed in the granulosa and the internal theca of the vitellogenic follicles. No immunoreactivity was observed in immature follicles. Using human POMC cDNA as the hybridization probe POMC-like mRNA was identified in situ in cells of the granulosa and internal theca of the vitellogenic follicles. No labeling was observed in primordial follicles. The demonstration in the same cells of POMC mRNA and POMC-related peptides immunoreactivity indicates a local production of the opiate hormones.