Dipartimento di Scienze Biomediche Università degli Studi di Sassari viale San Pietro, 43/C 07100 Sassari Italia
bandiera@uniss.it
Prof. Pasquale Bandiera is born in Pozzomaggiore (SS) on March 15, 1961, Graduated at Medical Faculty of the Sassari University. Position: Associate Professor of Human Anatomy (BIO16) at the Department of Biomedical Sciences, Sassari University. Professional Background: applicant for general medical service from 1993 to 1995; Researcher, Medical Faculty at University of Sassari since 1995; Confirmed Researcher, since 1998; Associate Professor, Medical Faculty University of Sassari, from 2006. Courses and educational activities: Professor of Anatomy at Medical Faculty, Bachelor Courses of Nursing and Neurophysiopathology Technician; Professor of Anatomy at Pharmacy Faculty, Bachelor Courses of Herbalist and Toxicology of Environmental Pollutants. Conduct integrative seminars and lessons in official courses of Anatomy for Medical and Dentistry students. Coordination: Coordinator of Laboratory of Biology of Reproduction and Development at Department of Biomedical Sciences. It coordinates and conducts exercises of Osteoarthrology and macroscopic and microscopic Anatomy for Medical and Dentistry students. Scientific field of interest: component of national and international research team, responsible for research funds at a local level. Research activities in various fields of human Anatomy: in particular Biology of Reproduction by studies on the ultrastructural morphology of human sperm in infertile subjects. Scientific Society: It. Soc. Anatomy since 1986, It. Soc. Experimental Biology since 1992, It. Soc. Andrology since 1997, It. Soc. of Electron Microscopy and European Microscopy Society since 1999, Italian Group of Cytometry since 2000, Italian Society of Toxicology of Reproduction since 2006.
Abstract: Ultrasound-targeted microbubbles (MBs) offer new opportunities to enhance the capabilities of diagnostic ultrasound (US) imaging to specific pathological tissue. Herein, we report on the design and development of a novel prototype of US contrast agent based on polymeric MBs targeted to prostate-specific membrane antigen (PSMA) for use in the diagnosis of prostate cancer (PCa). First, a set of air-filled MBs by a variety of biocompatible polymers were prepared and characterized in terms of morphology and echogenic properties after exposure to US. MBs derived from poly(d,l-lactic-co-glycolic acid) (PLGA)-poly(ethylene glycol) (PEG) copolymer resulted as the most effective in terms of reflectivity. Such polymer was therefore preconjugated with a urea-based PSMA inhibitor molecular probe (DCL), and the obtained MBs were investigated in vitro for their targeting efficacy toward PSMA positive PCa (LNCaP) cells. Fluorescence microscopy proved a specific and efficient adhesion of targeted MBs to LNCaP cells. To our knowledge, this work reports the first model of polymeric MBs appropriately engineered to target PSMA, which might be further optimized and used for PCa diagnosis and potential carriers for selective drug delivery.
Abstract: Molecular targeted cancer therapy mediated by nanoparticles (NPs) is a promising strategy to overcome the lack of specificity of conventional chemotherapeutic agents. In this context, the prostate-specific membrane antigen (PSMA) has demonstrated a powerful potential for the management of prostate cancer (PCa). Cancer chemoprevention by phytochemicals is emerging as a suitable approach for the treatment of early carcinogenic processes. Since (-)-epigallocatechin 3-gallate (EGCG) has shown potent chemopreventive efficacy for PCa, we designed and developed novel targeted NPs in order to selectively deliver EGCG to cancer cells. Herein, to explore the recent concept of "nanochemoprevention", we present a study on EGCG-loaded NPs consisting of biocompatible polymers, functionalized with small molecules targeting PSMA, that exhibited a selective in vitro efficacy against PSMA-expressing PCa cells. This approach could be beneficial for high risk patients and would fulfill a significant therapeutic need, thus opening new perspectives for novel and effective treatment for PCa.
Abstract: Development of molecules chemically modifying the expression of crucial orchestrator(s) of stem cell commitment may have significant biomedical impact. We have recently developed hyaluronan mixed esters of butyric and retinoic acids (HBR), turning cardiovascular stem cell fate into a high-yield process. The HBR mechanism(s) remain still largely undefined.
Abstract: We report a 12-year-old patient with Patau syndrome, in whom two cell lines were present from birth, one with total trisomy 13 due to isochromosome (13q), and one with partial trisomy 13. A cytogenetic re-evaluation at 9 years of age brought to light in skin fibroblasts a third cell line, partially monosomic for chromosome 13. The derivatives (13) present in the three cell lines were characterized through fluorescence in situ hybridization (FISH) experiments with suitable probes; the results suggested a sequence of rearrangements which beginning from an isochromosome (13q) could have led to the other two derivatives. We report the clinical data at birth and at the age of 12; at this age pigmentary lesions with phylloid pattern were noted. Cytogenetic findings of the chromosomal analyses on different tissues, including skin fibroblasts from differently pigmented areas, are also reported.
Abstract: We describe two malformed infants with trisomy 6p12.1-p22.1 due to 12/6 interchromosomal insertion. The phenotypic data observed in these patients are compared chiefly with a case cytogenetically similar described by Villa et al. [A. Villa, E.G. Gomez, L. Rodriguez, R.H. Rastrollo, M.E. Martinez Tallo, M.L. Martinez-Frias, Interstitial tandem duplication of 6p: a case with partial trisomy (6)(p12p21.3), Am. J. Med. Genet. 90 (2000) 369-375]. All three infants are trisomic for a genomic segment which largely overlaps that reported as duplicated in previous cases, but with the addition of a more proximal segment, extending from 6p12 to 6p21. We suggest that some of their phenotypic anomalies are due to the trisomy of this chromosomal region. We also speculate on the possible role played by the TFAP2B (Transcription Factor AP2-beta) gene, which is one of the genes mapped on the duplicated segment.
Abstract: The histopathology of the central parts of 40 A1 pulleys from adult patients with primary trigger fingers was studied using light and transmission electron microscopes and the findings were compared with those in a control series of 10 normal A1 pulleys. The evaluation of the normal A1 pulley revealed a bi-laminar structure. The deepest layer was composed of dense normal connective tissue. The outermost layer was formed by loose connective tissue. In trigger digits, it was possible to identify a tri-laminar structure. The deepest layer was composed of irregular connective tissue, formed by small collagen fibres and abundant extracellular matrix. A considerable amount of chondroid-metaplasia was present in this layer. The middle layer contained dense, normal connective tissue with some fibrocytes. The outermost layer was formed of loose connective tissue. In conclusion, there was an additional layer in the A1 pulley in pathological cases which was not present in normal pulleys.
Abstract: We report the results of a molecular study of a large family segregating the complete form of the Androgen Insensitivity Syndrome (CAIS) in several family members from three generations. We identified the mutant allele by polymerase chain reaction (PCR) amplification of the short tandem repeat (CAG)n, highly polymorphic in the population, present in the first exon of the androgen receptor (AR) gene. In this family four different alleles were detected and one of these showed a perfect segregation with the disease. This study enabled us to identify the heterozygous females in this family. We think that this simple, indirect test, is also suitable for prenatal diagnosis of Morris' syndrome when the mother is heterozygous for the size of the short tandem repeat and one affected subject in the family may be studied.
Abstract: We report the results of a molecular study of a large family segregating the complete form of the Androgen Insensitivity Syndrome (CAIS) in several members from three generations. We identified the mutant allele by Polymerase Chain Reaction (PCR) amplification of the short tandem repeat (CAG)n, highly polymorphic in the population, present in the first exon of the androgen receptor (AR) gene. In this family four different alleles were detected and one of these showed a perfect segregation with the disease. This study enabled us to identify the heterozygous females in this family. We think that this simple, indirect test, is also suitable for prenatal diagnosis of Morris' syndrome when the mother is heterozygous for the size of the short tandem repeat and one affected subject in the family may be studied.
Abstract: The mechanism of formation of the maxillary sinuses is not elucidated as yet, although their morphology during embryogenesis is well described. In the prenatal period, the pneumatization hypothesis is not valid. As the molecular approach to this problem is difficult to apply to human samples, we decided to apply immunohistochemical reactions to analyse the synthesis of selected molecules involved in the rebuilding of tissues. Hematoxylin-eosin staining and immunohistochemical reactions for the detection of MMPs (matrix metalloproteinases), one of their inhibitor TIMP 1 (tissue inhibitor of MMPs), BMP 6 (bone morphogenetic protein 6) and TGF-beta (transforming growth factor beta) were performed in the epithelium the mucosa of the maxillary sinuses of several human foetuses from the collection of the Anatomical Institute. The age of the foetuses was 8, 11, 15, 16, 17, 18 and 22 weeks. An intense positive reaction for MMPs 1, 2 and 3 was found in the mucosal epithehum of developing sinuses in the whole series of foetuses was found. The reaction was more intense in advanced stages of foetal development. Tissue derived inhibitor TIMP was hardly detectable, regardless of the age of samples. However, the intensity of the reaction for TGFbeta was strong in both young and more mature sinus epithelium. The presence of BMP 6, a member of the superfamily of TGFbeta, was detected although the intensity of this reaction in the epithelium was rather weak. Both TGFbeta and BMP 6 are well known as regulators of differentiation in the course of organogenesis. Results of the histochemical analysis suggest the possible involvement of the epithelium in the growth and formation of the maxillary sinuses. The main argument for this is intense reaction for MMP proteases which, as in bone, regulate the turnover and rebuilding processes of the extracellular matrix (ECM).
Abstract: In this paper we tried to analyse in quantitative terms the differences in trabecular structure of human mandibles 8, 10 and 12 weeks old. The analysis was performed on decalcified Sirius-red stained trabeculae, photographed in polarised light. The data obtained from Fourier transforms of scanned pictures were evaluated by discriminant analysis to show the differences between the analysed stages of development. Also, various parts of the mandible were compared because the clinical experience shows that the frequency of pathological changes is different in different parts of mandible bone. The complexity of trabecular structure of bone samples was measured by fractal dimension. It is concluded that this quantitative approach is reliable and, in the future, could be used for physiological and pathological analyses of bone biopsies.
Abstract: This morphological study demonstrates that during the early phases of pregnancy, in the syncytium there are individual multinucleated units joined to one another by junctional complexes which probably are destined to flow together with the main syncytium by membrane breakdown. Normal human placentas from 6th to 12th week of pregnancy, derived from voluntary abortions, have been utilized. According to the method described by Tedde et al. (1991), clusters of villi have been isolated, washed in saline and subjected to a treatment with a 0.3% trypsin solution that caused the detachment of numerous cellular elements from the surface of the free villi. Such isolated elements have been subjected to centrifugation and separation in Percoll gradients. Then multinucleated syncytial units have been collected, attached to slides by cytocentrifugation, stained with toluidine blue, observed at the light microscope and utilized to count their nuclei: about 600 elements from different weeks of pregnancy have been considered and the results have been submitted to statistical analysis. The svncytial units are multinucleated and can often be different from one another, particularly in regard to the characteristics and the number of the nuclei. The evaluation of the number of nuclei contained in different syncytial elements shows that nuclei are less numerous in the early phases (6-8 weeks) and increase in number towards the end of the first trimester in a statistic significant way. In conclusion, the present study could confirm the existence of single syncytial units, joined to constitute the syncytial layer, probably destined to a complete fusion in the main syncytium and likely in functional relationship with the underlying Langhans cells.
Abstract: Bone collagen has a specific molecular ultrastructure which can be proved by birefringence. This protein, forming the main organic component of bone tissue, is known to survive millennia in paleontological bones and teeth. Birefringence of bone collagen obtained from the skeletons of the Nuraghi population living in Sardinia c-ca 1500 years B.C. was found previously by the use of polarizing microscopy [1]. In this paper, using high pressure liquid chromatography (HPLC) techniques, we show the existence of bone collagen cross-links preserved in Nuraghi skeletons after more than 3000 years.
Abstract: Dating of skeletons of Nuragic population living in Sardinia island centuries BP, based on the quantitative evaluation of the concentration of stable paramagnetic species produced by ionising radiation in tooth enamel was performed by using EPR spectrometry. Applying the additive dose method (60Co gamma rays) and comparative calculations based on analogous measurements done with Roman skeleton of the known age as discovered close to Nuragic tomb (Tombs of Giants, La Testa S. Teresa di Gallura, Sardinia) the age of Nuragic skeleton was evaluated as equal to about 3,200 years (1,200 years BC). The total error of EPR measurements and dose extrapolation was estimated for 10-12%. Crystallinity of bone mineral in Nuragic skeleton evaluated by the EPR technique, adapted earlier by some of the authors of the present paper for biomedical studies on mineralised tissues, is only little changed after the centuries of its deposition in the tomb when compared with contemporary bone tissue. Comparison of chemical composition of Nuragic skeleton contaminated through slow percolation by rain of floods with that of contemporary bone sample shows the increase of the concentration of Fe, PO4, SiO2, Al and Mg, Fe, SiO2, AP are not present in contemporary bones. As expected, the contamination was minimal in tooth enamel.
Abstract: The evaluation of the volume of the encephalic ventricles in the living human has a great importance in diagnosis and monitoring of several pathologies. As well as a good measuring accuracy, a great usefulness has the facility of execution. The present study would be a contribution to the search of systems of analysis that assure both fast execution and greatest precision of the quantitative values. Brain MRI scans from 10 female individuals, aged from 25 to 60, with no apparent neurologic or psychiatric pathology have been studied. The brains were evaluated with 5 mm slices thickness (gap 1 mm) scans performed in axial and coronal planes and obtained by means of long and short spin echo sequences. Images have been acquired on a Personal Computer and submitted to a semiautomatic morphometric analysis. The values obtained are in accordance with the range of normal values reported from the literature, showing also a lower standard deviation; besides this clear accuracy of measurement, it's important to emphasize the facility and swiftness in the execution of the imagine evaluation.
Abstract: The authors have carried out a study on the immunitary mechanisms which stimulate and avoid eventual alterations of infected periapex. Above all the aim of this first study has been the microscopic and ultrastructural valuation of the cellular components that characterize the process of chronic phlogosis of periradicular tissue, lymphocytes T and B, plasmacells and macrophages, and of those even more typical of the soft reactive tissues, fibroblasts and epithelial cells. It's just the interaction among these immunocompetent cells which determines the structural change of the periapical bone whose most common image of radiotransparence make it possible to diagnose the sufference of the pulpo-periapical system.
Abstract: The molecular structure of bone collagen was found to be preserved in Nuragic bone samples several centuries after burial. Enhanced birefringence of bone and dentin collagen fibres observed in polarized light after Sirus red staining was used to demonstrate this fact.
Abstract: The size of the aortic arch and thoracic aorta was measured in 50 healthy volunteers studied by NMR. In each case the following parameters were measured and statistically elaborated: diameter (in mm) of ascending aorta (between onset and arch beginning), of aortic arch (intermediate zone), of isthmus, of descending aorta (between isthmus and diaphragm), of diaphragmatic isthmus; a mean diameter was also calculated, and for every diameter mean and standard deviation were obtained. Moreover, the length (in mm) of superior and inferior contours of the aortic arch, as the ratio of these two values were obtained. At last, total length of considered aortic tract, area of visualized surface and theoretic volume were obtained. For each parameter a diagram was then plotted, with age in ordinates and values of above mentioned parameters in abscissae. Statistical evaluation allows to affirm that the size of the aorta grows with ageing. On the contrary, the ratio between superior and inferior contour of the aortic arch tends to remain unvaried; this fact seems to disagree with the current opinion that the superior wall of the arch grows with age more than the inferior one.
Abstract: The particular shape of the nasopharynx in the child can influence morphology and physiopathologic behaviour of the mucosa. In such a perspective, it appeared interesting to evaluate the kind and the entity of its variations by means of nuclear magnetic resonance (NMR) in 50 patients (age range: 2 months-16 years). The results show that the nasopharyngeal shape (represented either by bending radius and shape factor) varies in a statistically significant way during the considered age range, while the dimension of the pharyngeal tonsil remains constant in the studied cases. The statistically significant shape variation of the tonsil appears, so, related to its involvement in the shape variation of the whole nasopharynx.
Abstract: The cytopathic effects of Trichomonas vaginalis treated with inhibitory concentrations of anticytoskeletal compounds (mebendazole, griseofulvin, colchicine, taxol, and cytochalasin B) were studied in mouse CLID fibroblast cultures. The evaluation, at different times, of cell numbers and morphological alterations showed that cytopathic effect was considerably reduced when protozoa were pretreated with mebendazole and griseofulvin, whereas colchicine, taxol, and cytochalasin B had less effect. Furthermore, treatment with mebendazole, griseofulvin, and colchicine decreased adhesiveness of the protozoan, whereas treatment with cytochalasin B and colchicine completely inhibited its phagocytic activity. From these results it may be concluded that alterations induced in the trichomonal cytoskeleton may affect its adhesiveness and its in vitro cytopathic effect, but there is no direct correlation between protozoan phagocytosis and its in vitro pathogenic effect.
