Fourier Transform Infrared Imaging analysis of dental pulp inflammatory diseases.

OBJECTIVES: Fourier Transform Infrared microspectroscopy let characterize the macromolecular composition and distribution of tissues and cells, by studying the interaction between infrared radiation and matter. Therefore, we hypothesize to exploit this analytical tool in the analysis of inflamed pulps, to detect the different biochemical features related to various degrees of inflammation. MATERIALS AND METHODS: IR maps of 13 irreversible and 12 hyperplastic pulpitis, together with 10 normal pulps, were acquired, compared with histological findings and submitted to multivariate (HCA, PCA, SIMCA) and statistical (one-way ANOVA) analysis. The fit of convoluted bands let calculate meaningful band area ratios (means ± s.d., P < 0.05). RESULTS: The infrared imaging analysis pin-pointed higher amounts of water and lower quantities of type I collagen in all inflamed pulps. Specific vibrational markers were defined for irreversible pulpitis (Lipids/Total Biomass, PhII/Total Biomass, CH2 /CH3 , and Ty/AII) and hyperplastic ones (OH/Total Biomass, Collagen/Total Biomass, and CH3 Collagen/Total Biomass). CONCLUSION: The study confirmed that FTIR microspectroscopy let discriminate tissues' biological features. The infrared imaging analysis evidenced, in inflamed pulps, alterations in tissues' structure and composition. Changes in lipid metabolism, increasing amounts of tyrosine, and the occurrence of phosphorylative processes were highlighted in irreversible pulpitis, while high amounts of water and low quantities of type I collagen were detected in hyperplastic samples.

Does cyclic guanosine monophosphate induce autophagy in thyroid malignant carcinoma through down-regulation of cyclic guanosine monophosphate phosphodiesterase?

The aim of this study was to evaluate whether or not the expression of cGMP- phosphodiesterases (cGMP-PDE) varies in different thyroid pathologies and to elucidate the relationship between the expression of cGMP-PDE, cGMP, and autophagy. Fifty-four thyroid biopsy samples, excised to perform the biopsy, were split into two parts and randomly assigned: one part was microscopically examined and histological classified, and the other was frozen and analysed in order to evaluate the cGMP-PDE activity. Intracellular cGMP was also measured. A strong expression of intracellular cGMP and cGMP-PDE activity was observed in carcinoma in respect to controls and benign pathologies. The level of cGMP-PDE in papillary carcinoma without lymph node involvement (N-) was approximately four-fold higher compared to those with lymph node invasion (N±). On the contrary, the cGMP was one and a half times higher in N± than N-. Our results are promising, although further epigenetical studies are needed to confirm this association. A correlation between the cGMP-degrading activity and the severity of thyroid pathology has been shown. The decrease of cGMP-PDE and the increase of cGMP in N± papillar carcinoma could be an autophagic stimulus, a defence mechanism of the body, against the cancer that is expanding and invading other tissues and organs.

Solid variant of keratocystic odontogenic tumour: report of a case.

This case report presents a solid variant of keratocystic odontogenic tumour (KCOT), a benign uni- or multicystic intraosseous tumour of odontogenic origin, with a characteristic lining of parakeratinized stratified squamous epithelium and potential inflammatory infiltrate. A 52-year old female patient discovered occasionally, in an orthopantomography done for other reasons, the presence of a radiolucent lesion. A 2-3 cm wide lesion with well-demarcated margins was present in the premolar region of the left hemimandible. Under local anesthesia a biopsy was done and the diagnosis of odontogenic keratocyst was confirmed by histopathological examination. Under general anesthesia, the lesion was easily shelled-out and completely enucleated. The lesion had not recurred after a 6 years follow-up. The occurrence of a solid variant of KCOT could strengthen the hypothesis of a neoplastic rather than cystic nature of this lesion.

Interferon-alpha therapy and anti-human leukocyte antigen antibodies in hepatitis C virus-positive patient: case report.

Interferon-alpha (IFN-alpha) is currently the only treatment for patients with chronic hepatitis C. Yet it can induce acute renal transplantation rejection possibly by stimulating humoral responses. We tested patient sera for detection of donor-specific anti-human leukocyte antigen (HLA) antibodies observing an increased panel-reactive antibodies value after IFN-alpha therapy. Then, we also investigated whether antiviral treatment with IFN-alpha was related to an increased and/or different production of class I and class II anti-HLA antibodies. Patient sera analysis performed by a cytofluorimetric method using flow PRA tests showed the appearance of new HLA-antibody specificities. This study underlined that INF-alpha therapy modifies a patient's immune profile; hence, it is recommended to confirm HLA-antibody specificities after treatment in order to protect recipients from enhanced rejection risk owing to a false-negative donor-specific cross-match.

CD1a and CD1e allele frequencies in an Italian population from the Abruzzo region.

CD1 is a small family comprising 5 MHC-like genes located on chromosome 1 and encoding glycoproteins termed CD1a, CD1b, CD1c, CD1d and CD1e. They are expressed mainly on the surface of dendritic cells, monocytes and some thymocytes and are specialized in presenting lipid antigens to T lymphocytes. The structure is similar to that of MHC class I molecules with 3 globular domains and the Beta2-microglobulin. It has been shown that all five human CD1 genes exhibit a limited number of polymorphisms in the alpha1 domain whose effects are still unknown. CD1e results to be the most polymorphic isoform with six CD1e alleles (01, 02 in exon 2 and 03, 04, 05, 06 in ex3) described to date. At this moment, few investigations on the allele frequencies of the CD1 genes have been reported and all additional information improves our knowledge on this new class of antigen-presenting molecules. In order to study possible allelic variations of exon 2 of human CD1a and CD1e genes, we analyzed, by a sensitive technique, the sequence-based typing (SBT), 114 DNA samples from unrelated healthy Italian individuals from the Abruzzo region. Our experimental findings indicate that the allele frequency distribution of both CD1a and CD1e genes is in accordance with that observed in other geographic areas and did not identify any new allele, thus confirming a very low polymorphism.

Genetic predisposition to rheumatoid arthritis in a Tuscan (Italy) ancient human remain.

Rheumatoid arthritis (RA) is currently believed to have originated in America, and after the discovery of this continent in 1492, to have been exported to the Old World. We evaluated the genetic predisposition to RA in the "Braids Lady" from Arezzo (Italy), a partially mummified woman's body dating back to the end of 1500 AD which presents the anatomical and pathological features of this disease. The study of the polymorphic HLA-DRB1 locus, which includes alleles strongly associated with RA onset, has received much attention over recent years, especially the loci codifying for the DR1 and DR4 antigens, widely represented in the Mediterranean population, and for DR14, widespread among Native Americans. Molecular analysis was performed on extracts of DNA from the mummy, firstly from histological bone sections and then from the whole bone. Two different HLA typing techniques, PCR-sequence-specific oligonucleotides (PCR-SSO) and PCR-sequence-specific primers (PCR-SSP), were employed to identify HLA-DRB alleles. Both genotyping methods showed that the "Braids Lady" carried the DRB1*0101 allele, the serological equivalent of the DR1 antigen. Although the possession of RA risk factor genes cannot be considered a diagnostic marker, the positive result of the Italian mummy for DRB1*0101 and the RA features present, support the idea that this pathology was present in the Old World from at least the mid-16th century. A pathogenetic hypothesis of RA which might well explain its worldwide diffusion is the "molecular mimicry", resulting from a cross-reactive antibody response between certain microbial antigens and shared epitopes of specific HLA-DR1, DR4 and DR14 susceptibility alleles, the frequency of which varies among different ethnic groups.

MIB-1, Bcl-2 and p53 in odontogenic myxomas of the jaws.

Odontogenic myxoma is a rare benign neoplasm occurring in the jaws. Microscopically, it is composed of spindle or stellate-shaped cells arranged in a mucinous matrix. In some cases (20%), odontogenic epithelial islands may be found. The Authors evaluated p53, MIB-1, and Bcl-2 expressed by the epithelial and stromal elements in 12 cases of odontogenic myxoma of the jaws. The cells of the odontogenic epithelium were positive for Bcl-2, p53 and MIB-1. The stromal cell showed a high positivity for MIB-1. Proliferation of both the epithelial and stromal components could be related to the growth of this odontogenic tumour.

cAMP phosphodiesterase activity evaluation in human carcinoma of salivary glands.

The aim of this study was to evaluate differences of cAMP-PDE activity in human salivary glands, between a control group and some different benign tumours groups and, where present, with 2 malignant tumors groups. The value of the enzymatic activity in the groups analysed was 50% lower than control samples. The differences between the control group (82 +/- 7.9 nmols/mg of protein) and the 3 pathologic groups (Benign A: 44 +/- 6.2; Malignant A: 40 +/- 16; Benign B: 40 +/- 14.2; Malign A: 9.1; Benign C: 22 nmols/mg of protein) are statistically significant.

Pleomorphic lipoma of the oral cavity. Report of a case.

Pleomorphic lipoma (PL) is a rare benign tumor mainly located in the upper back, upper shoulders, and back of the neck in elderly men. More rarely it is located in the head and neck region and in the oral cavity. The differential diagnosis should be made with sclerosing liposarcoma and well-differentiated liposarcoma. A 59-year-old male patient was referred for the presence of a lesion involving the marginal and adherent gingiva of teeth # 5; this lesion extended into the vestibular mucosa. The lesion had a 2 cm diameter, showed no tenderness, had a hard-parenchymatous consistency, was mobile on the underlying tissues and was covered by normal appearing mucosa. Under local anesthesia, the lesion was completely removed. A free gingival graft from the palate was used to cover the defect. Microscopically, it was possible to observe mature adipocytes, spindle cells and rare ''floret-like'' cells. Lipoblasts and mitoses were absent. The definitive pathologic diagnosis was pleomorphic lipoma. No recurrences were present after a 5 years follow-up. Local excision is adequate for PL and the tumor does not recur.

CD10 expression in stromal cells of oral cavity squamous cell carcinoma: a clinic and pathologic correlation.

OBJECTIVE: CD10 is expressed on the majority of follicle-center lymphomas and Burkitt lymphomas. CD10 has also been shown to be present in a variety of other neoplasms. DESIGN: The aim of this study was a correlation of CD10 and several parameters: age, tumor size, presence of lymph node metastases, clinic stage, histologic grading, presence of local recurrences. MATERIALS AND METHODS: The tissues of 77 consecutive patients with oral cavity squamous cell carcinoma were evaluated using immunostaining with monoclonal antibody for CD10. MAIN OUTCOME MEASURES: Highly significant correlations were found with the lymph node status, the presence of local recurrences and the histologic grading. The presence of CD10-positive cells was not correlated with the age of patients, tumor size and clinic stage. RESULTS: The results of the present study show that in oral squamous cell carcinoma CD10 positivity is an indicator of worse prognosis. Another strong correlation was found with the presence of local recurrences. Also the histologic grade was significantly correlated with the CD10 positivity. CONCLUSION: Our results point to the fact that CD10 expression can, perhaps, have an important role in tumor invasion, probably facilitating the occurrence of metastases.

Waiting list for kidney transplants.

Medical and technological progress have made kidney transplants an effective, alternative therapy to dialysis for patients suffering from chronic kidney failure. Transplantation improves the quality of life of these patients significantly; however, waiting lists are long and this is because of the attitude of the general public to organ donation, not a lack of medical expertise. In fact, the only limiting factor in kidney transplant is the opposition to donation expressed by the deceased or family members. Herein we outline the distribution of patients on the kidney transplant waiting list in the Regional Transplant Centre for Abruzzo and Molise in L'Aquila, Italy, and highlight the reasons why patients are withdrawn from the list, the main reason being a deterioration in patient condition after long periods of dialysis.

HLA DR/DQ alleles and risk of type I diabetes in childhood: a population-based case-control study.

The objective was to evaluate HLA DR/DQ alleles and their risk factor for type 1 diabetes in the Abruzzo region (central Italy). Sixty incident cases from the Abruzzo region were studied together with 120 unrelated control subjects living in the same administrative areas. The relative risk of diabetes associated with the alleles under study was calculated by deriving the odds ratio (OR) maximum likelihood estimates and their 95% confidence intervals (CI) by the exponentiation of the logistic regression beta-parameter. The combination DRB1*03/DQA1*0501/DQB1*0201 was found in 20.0% of patients and 7.1% of the control subjects, conferring an OR of 4.04 and a CI of 1.97-8.49. The combination DRB1*04/DQA1*0301/DQB1*0302 was found in 23.3% of diabetic patients and 6.7% of controls, giving an OR of 5.69 and a CI of 2.77-12.05. DRB1*11/DQA1*0505/DQB1*0301 and DQA1*0505/DQB1*0301 were negatively associated with type 1 diabetes (OR=0.27, CI 0.11-0.57; OR=0.07, CI 0.02-0.19). The DQA1 genotype at risk was found to be DQA1*0301/DQA1*0501: OR=23.80, CI 2.97-190.89, as it occurred with the highest frequency in the patient group. The DQB1 genotype at risk was found to be DQB1*0201/DQB1*0302, which occurred in 13.3% of patients but in only 1.1% of the control group (OR=29.75, CI 5.36-549.25). Our results shed further light on the risk of development of this disease during a specific time period in an area where the overall incidence of type 1 diabetes is known.

Immunohistochemical analysis of soft tissues in implants with healthy and peri-implantitis condition, and aggressive periodontitis.

Today, implant-supported prostheses are widely accepted as a reliable treatment modality, but failures in longitudinal studies have been shown. In some cases, peri-implantitis with a progressive periodontal bone loss takes place, and mechanical or load factors and biological or plaque-induced lesions have been claimed as main etiologic factors. We compared five cases of peri-implantitis, with five cases of healthy peri-implant tissues and five cases of aggressive periodontitis in order to give new findings on the osseointegration loss process. Biopsy specimens from the peri-implant tissues including oral (O), sulcular, and junctional epithelium and the underlying and supracrestal connective tissue, were taken in all cases for histological and immunohistochemical analysis. T lymphocytes were the most prominent cell in the peri-implantitis (PG) and aggressive periodontitis (AG) groups, but not in the peri-implant healthy group (HG). CD1a-positive cells (Langerhans and immature dendritic cells) were observed more frequently in the O than in the sulcular-junctional (S-J) epithelium: they were located in the basal and parabasal layers, without any differences between the three groups. Vascular proliferation analysed by immunoreactivity for CD34, Factor VIII, and vascular endothelial growth factor was more prominent in the PG comparing with HG and AG in the S-J area. Apoptosis, analysed by bcl2 and p53 immunoreactivity, was similar in the three groups. In conclusion, we suggest that the osseointegration loss process is due to an inflammatory process similar to that observed in aggressive periodontitis according to the number of T lymphocytes, but not to the vascular proliferation.

Transforming growth factor-beta 1 (TGF-beta 1) expression in normal healthy pulps and in those with irreversible pulpitis.

AIM: To evaluate the Transforming Growth Factor-beta 1 (TGF-beta 1) expression in normal healthy pulps and in those with irreversible pulpitis. METHODOLOGY: Twenty-three normal, healthy pulps were removed from mandibular third molars, and 20 pulps were retrieved from teeth with irreversible pulpitis. TGF-beta 1 was evaluated in the odontoblastic and subodontoblastic layers, in the stromal cells (fibroblasts), and in the blood vessels. TGF-beta 1 expression was determined by evaluating 500 cells in the odontoblastic and subodontoblastic layers and 500 fibroblasts in the stroma for each specimen, and counting the number of positive cells. The number of the positive vessels was evaluated in 10 high power fields (HPF). In almost all cases, the cellular positivity was cytoplasmatic. Statistical analysis was performed using Mann-Whitney U- and Student's t-tests. RESULTS: A higher expression of TGF-beta 1 was found in the odontoblastic-subodontoblastic layer of the irreversible pulpitis specimens; this difference was statistically significant (P = 0.0002). No statistically significant difference was observed between the two groups in TGF-beta 1 expression in the stromal cells (P = 0.54) or in the vascular component (P = 0.94). CONCLUSIONS: The higher and statistically significant expression of TGF-beta 1 found in the odontoblastic-subodontoblastic layer of irreversible pulpitis specimens may indicate a role for TGF-beta 1 in the dentinal repair processes after pulp inflammation.

Expression of transforming growth factor-beta 1 (TGF-beta 1) in odontogenic cysts.

AIM: To evaluate the positivity to transforming growth factor-beta 1 (TGF-beta 1) in different types of odontogenic cysts. METHODOLOGY: A total of 30 radicular cysts (RCs), 27 follicular cysts (FCs) and 28 odontogenic keratocysts (OKCs) were evaluated for immunohistochemical analysis of TGF-beta 1. TGF-beta 1 was evaluated in blood vessels, stromal cells (fibroblasts) and pluristratified squamous epithelium. TGF-beta 1 expression was determined by evaluating the number of positive elements. TGF-beta 1 expression was determined by evaluating 1000 cells in the pluristratified squamous epithelium (500 in the basal and parabasal layers, and 500 in the superficial layer) and 500 cells (the fibroblasts in the stroma) for each specimen, and counting the number of positive cells. The number of positive vessels was evaluated in 10 high power fields (HPF). The Chi-square test was used to evaluate differences between the two groups (RC + FC and OKC). A P-value <0.05 was considered to indicate statistical significance. RESULTS: A higher and statistically significant positivity was found in the basal-suprabasal epithelial layers (P=0.0011), superficial epithelium (P=0.053) and stromal cells (P=0.0002) of orthokeratotic and parakeratotic OKC as compared with RC and FC. CONCLUSIONS: These differences suggest that control of the cell cycle may be abnormal in orthokeratotic OKCs. These OKCs may have an intrinsic growth potential not present in other cyst types.

cGMP phosphodiesterase activity evaluation in human carcinoma of salivary glands.

The aim of this study was to evaluate differences of cGMP-PDE activity in salivary glands, between a control group and different benign tumour groups and, where present, with malign tumour groups. Endogen cGMP was evaluated too. The enzymatic reaction used the method of Spoto et al., with minor variations. The samples were organized in six groups: A (Adenolymphoma and Warthins tumour); B (Pleomorphic Adenoma); C (Basaloid Adenoma); D (Myoepitelioma). The control group was represented by healthy patients. In A and B groups, we have analyzed malign pathologies (Adenocarcinoma and Parotid Lymphoma) The benign tumours have more activity than controls, especially in Myoepitelioma (D) but with a decrement in the C group, which presents lower values of cGMP than the other three groups, where the concentration is similar. Between A and B groups, the activity values of malign tumours are similar, higher than controls and than the other benign pathologies, but not higher than in myoepitelioma. The cyclic concentration is similar for malign pathologies, with concentrations lower than controls, similar to Basaloid Adenoma (C).

Genetic expression profiling of six odontogenic tumors.

Odontogenic tumors are rare neoplasms arising from the odontogenic apparatus. We aimed to identify molecular characteristics associated with odontogenic tumorigenesis and malignancy. To this end, we investigated the expression level of human genes by using, for the first time in odontogenic tumors, the technique of expression profiling. Gene expression alterations common to all six odontogenic tumors were identified by the use of cDNA microarrays containing 19,000 human cDNAs. Statistical analysis on a subset of 4974 cDNAs present in the biopsies identified 506 distinct genes associated with the tumors (p-value < 0.01). Gene ontology analysis of the cellular processes which were differentially regulated in odontogenic tumors was accomplished by the use of a subset of 1409 annotated genes. Finally, 43 cDNAs differentiated the three malignant odontogenic tumors (ameloblastic carcinoma, clear cell odontogenic tumor, granular cell odontogenic tumor) from the three benign ameloblastoma biopsies (p < 0.01). The identified genes might help us better classify borderline odontogenic tumors.

Genetic portrait of malignant granular cell odontogenic tumour.

Odontogenic tumours are rare neoplasms whose classification is sometime controversial. Among these entities, granular cell odontogenic tumour (GCOT) is extremely rare and usually has a benign clinical behaviour. While the histogenesis of GCOT remains to be clarified, we documented the existence of a malignant counterpart of this neoplasm and proposed the name of malignant GCOT. Expression profiling by cDNA microarrays is a molecular technology that enables a global gene expression analysis. By using cDNA microarrays, we identified in malignant GCOT several genes with significantly differentially regulated genes when compared to non neoplastic tissues. These cancer specific genes include a range of functional activities: (1) transcription, (2) signaling transduction, (3) cell-cycle regulation, (4) apoptosis, (5) differentiation and (6) angiogenesis. In conclusion, we show that cDNA microarrays is a useful approach to investigate the biology of tumours. Moreover, this technology might lead to identification of gene targets for cancer therapy and to molecular classification of odontogenic tumours.