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.

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.

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.

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.

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.

Oxidation of Fe(II) horse heart cytochrome c by ultrasound waves.

In aqueous solution, acoustically induced cavitation produces the collapse of bubbles containing gas and water-vapor, producing free radicals by the homolysis of the water molecules. Generally, under these extreme physical conditions, the secondary and tertiary structures of the proteins result are altered and denaturation phenomena are often observed. This paper discusses the evidence that, in the presence of argon and in oxygen-free experimental environment, the reduced horse heart cytochrome c, instead of undergoing a denaturation process, is oxidized to ferric-cytochrome c. Kinetic and circular dichroism measurements performed after ultrasound irradiation at a frequency of 38 kHz are reported. A possible correlation between ultrasound induced molecular damage to the tertiary structure of the proteins and their own extension of helix content is also hypothesized.

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.

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.

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.

Differentiation of odontogenic keratocysts from other odontogenic cysts by the expression of bcl-2 immunoreactivity.

Odontogenic keratocysts (OKC) present an aggressive course with a marked tendency to recurrence. The epithelium of OKC is thought to have an intrinsic growth potential and has been shown to present a higher rate of proliferation as compared to other types of cyst. bcl-2 has a role in the extension of cell survival. The objective of the present study was to evaluate the bcl-2 protein expression in different odontogenic cysts. A total of 19 dentigerous cysts (DC), 20 radicular cysts (RC) and 14 OKC were used in the present study. DC and RC showed an almost complete negativity for bcl-2. OKC, on the other hand, presented in all cases a strong positivity in the cells of the basal layer, with, in most cases, more than 50% of the cells positive. This bcl-2 positivity of the basal layer of OKC could point to an abnormal control of the cell cycle. The bcl-2 protein overexpression could then produce an increase in the survival of the epithelial cells, and this increased lifespan could, in turn, lead to the peculiar aggressive growth pattern of OKC. Moreover the bcl-2 staining can be useful to differentiate OKC from other types of odontogenic cysts.

Solitary fibrous tumour of the tongue.

Solitary fibrous tumour (SFT) is a neoplasm most often localised in the pleura and peritoneum. The tumour is composed of spindled fibroblastic cells arranged in a haphazard way. Recently SFT has been described in many locations. Only one case of oral SFT has been described in the cheek: this is the second case of an oral SFT located in the tongue. The differential diagnosis must be made from many soft tissue tumours. SFTs stain strongly, in almost all cases, for CD34.

Angiomyolipoma of the palate. Report of a case.

Angiomyolipoma (AML) is a tumour or an hamartomatous growth that usually affects the kidney. Only rarely has AML been described in the oral cavity. The authors report a case of AML located in the palate in a 43-year-old patient. AML is composed of smooth muscle cells, blood vessels and mature fat cells. In 50% of cases, AML presents with symptoms of tuberous sclerosis. Renal AML are often invasive, may involve regional nodes and may recur, while, on the contrary, AML are most often well circumscribed and easily resected. AML seems to follow an entire benign course.

Sebaceous adenoma of the cheek.

Sebaceous adenoma is a tumour only rarely located in the oral cavity. Less than 10 cases have been reported. Sebaceous adenoma represents 0.5-0.7% of all monomorphic adenomas. Sebaceous adenoma is mainly constituted by two types of cells, undifferentiated peripheral basaloid cells and cells showing different degrees of sebaceous differentiation located in the center of the lesion. The differential diagnosis must be made with sebaceous hyperplasia. Sebaceous adenomas are benign, and they do not recur after a conservative excision.

Osteolipoma of the tongue.

Lipomas are common, benign tumours located in any part of the body in which fat is normally present. Some variants of lipoma have been described according to the type of tissue present. A rare variant consists of a lipoma with osseous or cartilaginous metaplasia. These lesions have been called chondrolipoma, osteolipoma, lipoma with chondroid or osseous metaplasia, lipoma with cartilaginous or osseous change, or ossifying lipoma. We present the case of an osteolipoma of the tongue in a 49-year-old female who was referred for a painless mass on the left lateral margin of the tongue, and present for about 8 years. Osteolipomas have been reported in middle-aged or elderly patients with a very long clinical history. These tumours tend to be large and to arise from the deep soft or subcutaneous tissues. The cartilage and bone is probably produced by metaplasia of fibroblasts in chondroblasts or osteoblasts. These lesions are benign and do not recur.

Warty carcinoma of the oral mucosa in an HIV+ patient.

The authors present the case of a 36-year-old HIV+ male patient with a 1-cm diameter papillary exophytic lesion of the right cheek. Microscopic examination showed a papillary epithelial neoplasm with invasion of the stroma in the peripheral part. Cellular and nuclear atypia were present in the superficial and in the deep layers of the neoplasm. An in situ hybridization for human papillomavirus (HPV) 6, 11, 16, 18, 31, 35 and 51 was performed. A focal positivity only for HPV 16 and 18 was present in koilocytotic cells of the most peripheral portion of the lesion. The microscopic definitive diagnosis was warty carcinoma of the cheek. No recurrence was observed at a 3-year follow-up.