Nuclear morphometry in canine acanthomatous ameloblastomas and squamous cell carcinomas.

The aim of this study was to evaluate whether morphometrical analysis can be of diagnostic value for canine acanthomatous ameloblastoma. We calculated, by means of an automated image analyser, some morphometric nuclear parameters, in particular: mean nuclear area (MNA), mean nuclear perimeter (MNP), maximum and minimum diameters (MDx and MDm) coefficient of variation of the nuclear area (NACV), largest to smallest dimension ratio (LS ratio), and form factor (FF), in 8 canine acanthomatous ameloblastomas, and we compared these morphometric data to those of 13 squamous cell carcinomas of canine gingiva. The results indicated a progressive increase of the MNA, NACV, MNP and MDm proceeding from acanthomatous ameloblastomas (MNA: 42.11+/-8.74; NACV: 28,36+/-7,23; MNP: 24.18+/- 2.68; MDm: 5.69+/-0.49) to squamous cell carcinomas (MNA:49,69+/-9,10; NACV: 30,89+/-7,75; MNP: 25.63+/-2.54; MDm: 6.64+/-0.73). On the contrary, the LS ratio and the FF resulted greater in acanthomatous ameloblastomas (LS ratio: 1,63+/-0,12; FF: 1,13+/-0,002) than in SCCs (LS ratio: 1,40+/-0,12; FF:0.91+/-0.38). Moreover, the MNA, MNP,MDx and MDm resulted similar (MNA: p=0.89; MNP: p=0,65; MDm: p=0,16; MDx: p=0,13) in a subset of four acanthomatous ameloblastomas with cellular atypia (MNA:49,01+/-6,88; MNP: 26,28+/-1,99; MDm: 6.08+/-0.41; MDx: 10.18+/-0.88) and in squamous cell carcinomas (MNA:49.69+/-9,10; MNP: 25.63+/-2.54; MDm: 6.64+/-0.73; MDx: 9.26+/-1.05). While the NACV values resulted higher in typical acanthomatous ameloblastoma (29,99+/-6,06) than in atypical acanthomatous ameloblastoma (26,74+/-8,84) and similar to those of the SCCs (30,89+/-7,75). These results seem to confirm that acanthomatous ameloblastoma is a malignant or potentially malignant lesion and emphasizes that nuclear morphometry analysis can be an useful diagnostic and prognostic method in canine oral pathology.

Follicle-like model by granulosa cell encapsulation in a barium alginate-protamine membrane.

Granulosa cells from bovine and porcine ovaries were cultured either in monolayer or in follicle-like barium alginate capsules for 6 days. Morphological investigation by electron scanning microscopy indicated that culture in a three-dimensional (3D) system allows self-organization of spherical-polyhedral shape cells. The luteinization index (progesterone:17beta-estradiol ratio) was significantly higher for monolayer cells than for the 3D cell culture system, confirming the results of morphological analysis and indicating more physiological growth. The encapsulated 3D culture system appears to be a promising way of obtaining in vitro maturation and development of follicles and oocytes.

Transient transfection of porcine granulosa cells after 3D culture in barium alginate capsules.

Three-dimensional culture systems in barium alginate capsules can be employed to maintain primary granulosa cells in an undifferentiated state for almost 6 days. This is due to a self-organization of cells in a pseudofollicular structure. The transfection of primary granulosa cells is a necessary condition when employing these culture systems for several purposes, for example as an in vitro toxicity test or the development of oocytes or zygotes. In this work, the feasibility of two transient transfection techniques (liposome-mediated and electroporation) was assessed in primary porcine granulosa cells after a 6-day culture in an artificial extracellular matrix (barium alginate membrane). Human recombinant green fluorescent protein was chosen as a molecular readout, and protein expression was assessed after 48 hours from transfection. Liposome-mediated transfection gave low transfection levels, with increasing yields from 2 to 12 microgDNA/ml of medium; the maximum percentage (85.7%) was reached at 12 microgDNA/ml of medium. Electroporation-mediated transfection yields were higher: the best results (81.7% of transfected cells) were achieved with two 50V pulses and 12 microg/ml DNA. The application of a single or double pulse (50V) at 4 mgDNA/ml gave negligible results. These results indicate that primary granulosa cell cultured in barium alginate capsules can be transfected by electroporation with high transfection yields.

Clinical utility of fractionating erythrocytes into "Percoll" density gradients.

Two rapid methods for fractionating the RBC into five or nine layers of increasing density are reported. These procedures have been used to monitor the decline of glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) activity during the process of red cell aging in normal subjects and in beta-thal carriers, to study transfused patients with G6PD and pyruvate kinase (PK) deficiency and to test the effects of inositol hexaphosphate (IHP) encapsulation on RBC subpopulations.

An ovine preimmune foetal model to study the effect of cellular therapies for myocardic diseases.

The ovine foetus is an ideal model for preclinical medical studies of cell therapies. It allows us to follow the behaviour of repairing cells inserted into a favourable physiological microenvironment in an animal species more closely related to humans than the rat or rabbit. In addition, the preimmune foetus is able to support cell engraftment by eliminating the problems of tissue rejection. Labelled fibroblasts were transplanted into the myocardium of preimmune foetuses, injecting through a disposable bowed mouth pipette into the left ventricular apex. Two weeks later, foetuses were isolated by laparotomy and each heart was collected and morphologically analyzed. No cases of abortion or postoperative complications in mothers or foetuses occurred. Macroscopically, the isolated hearts did not display any abnormality apart from a small petechia at the injection site. Tissue sections did not show any sign of acute tissue inflammation and viable labelled cells were easily identified among myocardiocytes. This model system guarantees a reduced damage in the engrafted tissue, a high survival and easy retrieval of the injected cells. The direct injection of cells into the preimmune ovine foetus myocardium can be satisfactorily performed to control tissue delivery and to reduce the risk of cell loss and dispersion.

Pre- and postsurgical psycho-emotional aspects of the orthognathic surgery patient.

The current study represents an attempt to examine psychologic changes, emotional impact, expectations, and satisfaction in patients before and after orthognathic surgery. Levels of presurgical anxiety, postsurgical depression, body concept, and all the important changes in physiologic functions were measured by 4 questionnaires, which were self-administered before and after surgery. The results of this study suggest that surgery does, in fact, produce improvements in self-esteem and body image (patient's evaluation of his or her facial attractiveness) and in mastication and speech, and therefore in their lifestyle. All patients experienced a medium to high level of presurgical anxiety, but no major problems after surgery.