Giovanni Francesco Spatola

Immunohistochemical expression and distribution of orexin, orphanin and leptin in the major salivary glands of some mammals

The aim of the study was to determine by immunochemistry the expression of leptin, orexin A and orphanin FQ in the major salivary glands (parotid, submandibular and sublingual) of rat, sheep and cow. These peptides, originally synthesized in central nervous system, adipose tissue and peripheral tissues including gastrointestinal tract, play an orexigenic (orphanin and orexin) or anorexigenic (leptin) roles in the intricate neuronal network appointed to the control of nutritional homeostasis. Peptide-specific immunoreactivity was present in the studied salivary glands with various intensities in different species, in the ductal epithelium, sometimes in the acinar epithelium, and in nervous trunks spread in connective tissue stroma. The obtained data show that salivary glands present an unexpected source of orexigenic and anorexigenic peptides which with their autocrine, paracrine, and endocrine mechanisms of action may participate in the control of salivary gland function.

MMP-2, MMP-9, and iNOS expression in human dental pulp subjected to orthodontic traction.

OBJECTIVE To test the hypothesis that some metalloproteinases (MMP-2, MMP-9) and inducible nitric oxide synthetase (iNOS) enzymes in dental pulp samples do not vary when subjected to orthodontic treatment. MATERIALS AND METHODS Human dental pulps were taken from male and female patients (N=10; age 10-14 years). A straight wire technique was used with nickel-titanium or steel archwires. The increase of pressure applied on teeth was gradual. Five patients were subjected to premolar extractions after 14 months of treatment and one after 24 months. Samples were Bouin-fixed, paraffin-embedded, and afterwards processed for immunohistochemistry using anti-MMP-2, anti-MMP-9, and anti-iNOS antibodies. RESULTS A reduction of MMP-2, MMP-9, and iNOS expression occurred in treated samples. This became more evident with increased treatment time. CONCLUSION The hypothesis is rejected. The reduction of expression of those proteins revealed a time-dependent relationship.

Modulation of MMP-2 and MMP-9 in Churg-Strauss syndrome respiratory mucosa: potential monitoring parameters

Churg-Strauss (CSS) syndrome is rare and of unknown etiology. It is associated with vasculitis, blood eosinophilia and granulomatosis, and affects multiple organs and systems at various stages of the disease. Specific diagnostic and monitoring tests are not yet available. This study aims to assess the changes in MMP-2 and MMP-9 along with the histopathological alterations in two cases of CSS, as possible potential diagnostic and monitoring criteria. Two adult male patients were diagnosed with CSS in the otorhinolaryngology clinic in the University of Palermo, based on multiple clinical and histopathologic criteria. Biopsies of respiratory mucosa were taken after the consent of the patients, processed for routine histopathology and immunohistochemistry as well as quantitative polymerase chain reaction (qPCR). Similar biopsies were also taken from a non-CSS patient. The Assessment of MMP-2 and MMP-9 was performed using both immunohistochemistry and qPCR techniques. Histopathological alterations in the respiratory mucosa were consistent with vasculitis and granulomatous tissue formation, in addition to inflammatory cell infiltration with abundance of eosinophils. Immunohistochemistry assay performed on the samples derived from the two CSS patients showed a relative and remarkable increase of both MMP-2 and MMP-9 compared to controls. Such an increase was consistent with the qPCR results which depicted a significant increase between 20 and 30% for both MMP-2 and MMP-9, respectively. Since the secretion of MMPs is an essential step in angiogenesis, could these enzymatic factors be used as parameters to diagnose or monitor the evolution of CSS? The small number of samples analyzed in this study does not allow us to suggest a general statement correlating the increase in expression of MMP-2 and MMP-9 to the appearance or evolution of vasculitis; it is only speculative.

Immunohistochemical expression of apoptotic factors, cytokeratins, and metalloproteinase-9 in periapical and epithelialized gingival lesions

Bellmann K, 2010, CELL STRESS CHAPERON, V15, P101, DOI 10.1007-s12192-009-0126-9; Cappello Francesco, 2011, Front Biosci (Schol Ed), V3, P341, DOI 10.2741-s155; Cappello F, 2006, CANCER, V107, P2417, DOI 10.1002-cncr.22265; Cappello F, 2002, EUR J HISTOCHEM, V46, P199; Carneiro E, 2009, ORAL SURG ORAL MED O, V107, P127, DOI 10.1016-j.tripleo.2008.07.030; Chandra D, 2007, J BIOL CHEM, V282, P31289, DOI 10.1074-jbc.M702777200; Fujita Y, 2011, ODONTOLOGY, V100, P215; Garcia Celia Carrillo, 2007, Med Oral Patol Oral Cir Bucal, V12, pE585; Garcia CC, 2009, ORAL SURG ORAL MED O, V107, pE43, DOI 10.1016-j.tripleo.2008.12.002; Gregory CD, 2011, J PATHOL, V223, P177, DOI 10.1002-path.2792; Gupta S, 2002, CIRCULATION, V106, P2727, DOI 10.1061-01.CIR.0000038112.64503.6E; Hajra KM, 2004, APOPTOSIS, V9, P691, DOI 10.1023-B:APPT.0000045786.98031.1d; Kimi K, 2001, J ORAL PATHOL MED, V30, P434, DOI 10.1034-j.1600-0714.2001.300709.x; Kirkeby S, 2005, J IMMUNOL METHODS, V301, P102, DOI 10.1016-j.jim.2005.04.006; Lehr HA, 1997, J HISTOCHEM CYTOCHEM, V45, P1559; Leonardi R, 2003, J ENDODONT, V29, P180, DOI 10.1097-00004770-200303000-00004; Li P, 1997, CELL, V91, P479, DOI 10.1016-S0092-8674(00)80434-1; Mak BC, 2008, J BIOL CHEM, V283, P23462, DOI 10.1074-jbc.M803194200; Martins CA, 2011, J ENDODONT, V37, P36, DOI 10.1016-j.joen.2010.09.010; MARTON IJ, 1993, INT ENDOD J, V26, P131, DOI 10.1111-j.1365-2591.1993.tb00555.x; MOLL R, 1982, CELL, V31, P11, DOI 10.1016-0092-8674(82)90400-7; Nagata S, 1999, ANNU REV GENET, V33, P29, DOI 10.1146-annurev.genet.33.1.29; Pham NA, 2007, DIAGN PATHOL, V27, P2; Pritlove-Carson S, 1997, Oral Dis, V3, P19; Samali A, 1999, EMBO J, V18, P2040, DOI 10.1093-emboj-18.8.2040; Sawaf M H, 1991, J Biol Buccale, V19, P187; Skaland I, 2008, J CLIN PATHOL, V61, P68, DOI 10.1136-jcp.2007.046763; Skaland I, 2008, APPL IMMUNOHISTO M M, V16, P185, DOI 10.1097-PAI.0b013e318059c20c; Suzuki T, 2005, J ORAL PATHOL MED, V34, P46, DOI 10.1111-j.1600-0714.2004.00248.x; Suzuki T, 2002, J ORAL PATHOL MED, V31, P488, DOI 10.1034-j.1600-0714.2002.00016.x; Taylor CR, 2006, HISTOPATHOLOGY, V49, P411, DOI 10.1111-j.1365-2559.2006.02513.x; Tuan RS, 2011, INT J ORAL MAX IMPL, V26, P50; Tuan RS, 2011, INT J ORAL MAX SURG, V26, P63; Walker RA, 2006, HISTOPATHOLOGY, V49, P406, DOI 10.1111-j.1365-2559.2006.02514.x; Xanthoudakis S, 1999, EMBO J, V18, P2049, DOI 10.1093-emboj-18.8.2049

CB1 receptor expression in pancreatic islet of the obese and diabetes Zucker rats

The endocannabinoid system plays a key role in energy homeostasis, with agonists and antagonists of CB1 receptors acting centrally to stimulate and inhibit food intake, respectively. The currently available literature, about islets cannabinoid receptors expression and function, is confusing and often contradictory. Aim of our study is compare the expression of CB1 receptors in normal and obese Zucker (ZDF) rats.12 Male ZDF (6 lean and 6 fa/fa) aged 8 weeks were obtained from Harlan Italy S.r.l. Each rat was fed with standard diet and unlimited water. Specimens were taken at the age of 8-12-16 weeks after birth. Pancreas samples were fixed in formaline and embedded in paraffin. Section were processed immunohistochemically using DakoCytomation EnVision kit for anti CB1antibody (Biosource Europe S.A.). Leica microscope, with a DSL2 Nikon was used to observe in double blind. Images were submitted at the image analysis tools of the Photoshop CS5Ex. At the same time pancreas specimens were frozen in liquid nitrogen and stored at -80°C until use. A mRNA was extracted for all samples and a RT PCR was performed. The pancreatic islets cells in fa/fa show deep apoptotic alterations. Our observations show a CB1 expression extended to most of the cell population (eighth week), up to a structural disorganization in the sixteenth week; the expression of CB1 receptor is upregulated in fa/fa compared to lean. These results according to the involvement of endocannabinoid system in the disregulation of the food intake mechanism and a prevalence of this system in the obesity.

Immunohistochemical expression of Cytokeratins in periapical lesions

Objective. This study seeks to clarify the histological origin of apical lesions, through the expression of certain cytokeratins.Study design. In 3 years 30 patients were selected. After clinical and radiological exams were chosen patients who had severe apical lesions and tooth decay with exposure of the pulp chamber left untreated for very long periods (more the 12 months). Serial slides were prepared both for immunohistochemistry and Hematoxylin- Eosin. Material and methods. Twenty specimens coming from the 30 patients were used for our purpose. All periapical lesions were surgically extracted and fixed in Bouin mixture, and embedded in paraffin. Samples were processed immunohistochemically employing the instructions of the Envision Dako cytomation kit. Monoclonal antibobies against Cytokeratins (CKs) 1, 5 , 8, 10 and 14 were used. Slides were observed with Leica Laborlux-S microscope and the image were acquired with NIKON DSL2 System. Each sample has been analyzed with a “double-blind” system by two different operators. Moreover, the digital images acquired have been compared to an image analysis tools by Adobe photoshop CS5 Extended. Results. CKs 1, 5 and 8 are expressed in all samples at external epithelial layer with no statistically significant differences. CK 10 is weakly expressed in some epithelial cells lining the periapical lesion. CK 14 was negative in most samples, and little positive in some specimens. Conclusion. Our results suggest that most of the apical lesions we studied have an epithelial origin. The absence of CK14 positivity may be explained by the fact that during the extirpation of the lesion the basal epithelium remained attached to the surrounding bone.