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Xu et al. (2020) suggest that salivary glands are a potential target of severe acute respiratory syndrome–coronavirus 2 (SARS-CoV-2) infection, since the expression of angiotensin I–converting enzyme 2 (ACE2), the receptor needed for virus entry into host cells, was higher in salivary glands as compared with lung when the GTEx portal was queried.

It has been shown that stem cells are able to calcify both in vitro and in vivo once implanted under the skin, if conveniently differentiated. Nowadays, however, a study on their efficiency in osseous regeneration does not exist in scientific literature and this very task is the real aim of the present experimentation. Five different defects of 6 mm in diameter and 2 mm in depth were created in the calvaria of 8 white New Zealand rabbits. Four defects were regenerated using 2 different conveniently modified scaffolds (Bio-Oss® Block and Bio-Oss Collagen®, Geistlich), with and without the aid of stem cells. After the insertion, the part was covered with a collagen membrane fixed by 5 modified titan pins (Altapin®). The defect in the front was left empty on purpose as an internal control to each animal. Two animals were sacrificed respectively after 2, 4, 6, 10 weeks. The samples were evaluated with micro-CT and histological analysis. Micro-CT analysis revealed that the quantity of new bone for samples with Bio-Oss® Block and stem cells was higher than for samples with Bio-Oss® Block alone. Histological analysis showed that regeneration occurred in an optimal way in every sample treated with scaffolds. The findings indicated that the use of adult stem cells combined with scaffolds accelerated some steps in normal osseous regeneration.

Mesothelioma is a malignant tumor mainly correlated to occupational asbestos exposure. Rare reports describe its occurrence also in animals, mainly linked to asbestos in the environment. Asbestos exposure is demonstrated by the appearance of characteristic histological hallmarks: asbestos containing ferruginous bodies that are iron-based structures forming around fibers and also other dust particles. Here we present a clinical case of a suspect of mesothelioma in the peritoneum of a dog with parallel histological observation of ferruginous bodies. To possibly correlate the dog tumor to environmental exposure, we performed X-ray fluorescence (XRF) analyses at two different synchrotrons to resolve the ferruginous bodies' composition. While the histological examination diagnoses a tubulo-papillary mesothelioma, the XRF analyses show that ferruginous bodies contain Si particles, resembling formations of exogenous origin; however, the morphology is unlikely that of asbestos fibers. We speculate that the peritoneal mesothelioma of this dog could be related to environmental exposure to non-asbestos material.

Carbon nanotubes (CNTs) are promising products in industry and medicine, but there are several human health concerns since their fibrous structure resembles asbestos. The presence of transition metals, mainly iron, in the fibres seems also implicated in the pathogenetic mechanisms. To unravel the role of iron at mesothelial level, we compared the chemical changes induced in MeT-5A cells by the exposure to asbestos (crocidolite) or CNTs at different content of iron impurities (raw-SWCNTs, purified- and highly purified-SWCNTs). We applied synchrotron-based X-Ray Fluorescence (XRF) microscopy and soft X-ray imaging (absorption and phase contrast images) to monitor chemical and morphological changes of the exposed cells. In parallel, we performed a ferritin assay. X-ray microscopy imaging and XRF well localize the crocidolite fibres interacting with cells, as well as the damage-related morphological changes. Differently, CNTs presence could be only partially evinced by low energy XRF through carbon distribution and sometimes iron co-localisation. Compared to controls, the cells treated with raw-SWCNTs and crocidolite fibres showed a severe alteration of iron distribution and content, with concomitant stimulation of ferritin production. Interestingly, highly purified nanotubes did not altered iron metabolism. The data provide new insights for possible CNTs effects at mesothelial/pleural level in humans.

Germline mutations of the oncosuppressor gene breast cancer 1-associated protein 1 (BAP1) were recently related to an autosomal-dominant tumor predisposition syndrome (BAP1-TPDS), characterized by uveal melanoma, malignant mesothelioma (MM), cutaneous melanoma, and other malignancies. The demonstration that BAP1 mutations are strongly associated with MM has provided a real breakthrough in the study of genetic predisposition in MM, that may explain why only a fraction of asbestos-exposed individuals go on to develop MM.To evaluate the possible role of BAP1 mutations in the epidemiology of sporadic MM, and their relationship with asbestos exposure, we determined the prevalence of germline BAP1 mutations by the Sanger method in a group of 29 asbestos-exposed patients, 21 of which were diagnosed with MM. They were residents of Trieste, a ship-building town in Northeast Italy with a very high incidence of mesothelioma.We identified non-obviously pathogenetic germline sequence variants of BAP1 in 3/29 patients and in 2/21 MM cases (10%).Non obviously pathogenic germline sequence variants of BAP1 were found. Nevertheless, limitations of predictive web tools allowed us to comment on some interesting peculiarities of our findings.

Background: Germline mutations of the oncosuppressor gene breast cancer 1-associated protein 1 (BAP1) were recently related to an autosomal-dominant tumor predisposition syndrome (BAP1-TPDS), characterized by uveal melanoma, malignant mesothelioma (MM), cutaneous melanoma, and other malignancies. The demonstration that BAP1 mutations are strongly associated with MM has provided a real breakthrough in the study of genetic predisposition in MM, that may explain why only a fraction of asbestos-exposed individuals go on to develop MM. Materials and Methods: To evaluate the possible role of BAP1 mutations in the epidemiology of sporadic MM, and their relationship with asbestos exposure, we determined the prevalence of germline BAP1 mutations by the Sanger method in a group of 29 asbestos-exposed patients, 21 of which were diagnosed with MM. They were residents of Trieste, a ship-building town in Northeast Italy with a very high incidence of mesothelioma. Results: We identified non-obviously pathogenetic germline sequence variants of BAP1 in 3/29 patients and in 2/21 MM cases (10%). Conclusion: Non obviously pathogenic germline sequence variants of BAP1 were found. Nevertheless, limitations of predictive web tools allowed us to comment on some interesting peculiarities of our findings.

Summary In this study, we have performed a morphological analysis of crocidolite fibres interaction with mesothelial cells (MET5A) by combining conventional electron microscopy with atomic force (AFM) and scanning near-field optical microscopy (SNOM). After 6-h exposure at a crocidolite dose of 5 μg cm−2, 90% of MET5A cells interact with fibres that under these conditions have a low cytotoxic effect. SEM images point out that fibres can be either engulfed by the cells that lose their typical morphology or they can accumulate over or partially inside the cells, which preserve their typical spread morphology. By using AFM we are able to directly visualize the entry-site of nanometric-sized fibres at the plasma membrane of the spread mesothelial cells. More importantly, the crocidolite fibres that are observed to penetrate the plasma membrane in SNOM topography can be simultaneously followed beneath the cell surface in the SNOM optical images. The analysis of SNOM data demonstrates the entrance of crocidolite fibres in proximity of nuclear compartment, as observed also in the TEM images. Our findings indicate that the combination of conventional electron microscopy with novel nanoscopic techniques can be considered a promising approach to achieve a comprehensive morphological description of the interaction between asbestos fibres and mesothelial cells that represents the early event in fibre pathogenesis.

Asbestos bodies are the histological hallmarks of asbestos exposure. Both conventional and advanced techniques are used to evaluate abundance and composition in histological samples. We previously reported the possibility of using synchrotron X-ray fluorescence microscopy (XFM) for analyzing the chemical composition of asbestos bodies directly in lung tissue samples. Here we applied a high-performance synchrotron X-ray fluorescence (XRF) set-up that could allow new protocols for fast monitoring of the occurrence of asbestos bodies in large histological sections, improving investigation of the related chemical changes. A combination of synchrotron X-ray transmission and fluorescence microscopy techniques at different energies at three distinct synchrotrons was used to characterize asbestos in paraffinated lung tissues. The fast chemical imaging of the XFM beamline (Australian Synchrotron) demonstrates that asbestos bodies can be rapidly and efficiently identified as co-localization of high calcium and iron, the most abundant elements of these formations inside tissues (Fe up to 10% w/w; Ca up to 1%). By following iron presence, we were also able to hint at small asbestos fibers in pleural spaces. XRF at lower energy and at higher spatial resolution was afterwards performed to better define small fibers. These analyses may predispose for future protocols to be set with laboratory instruments.

Human exposure to asbestos can cause a wide variety of lung diseases that are still a current major health concern, even if asbestos has been banned in many countries. It has been shown in many studies that asbestos fibers, ingested by alveolar macrophages, disrupt lung iron homeostasis by sequestering iron. Calcium can also be deposited on the fibers. The pathways along which iron and above all calcium interact with fibers are still unknown. Our aim was that of investigating if the iron accumulation induced by the inhaled asbestos fibers also involves calcium ions accumulation. Lung sections of asbestos-exposed mice were analyzed using an extremely sensitive procedure available at the synchrotron facilities, that provides morphological and chemical information based on X-ray fluorescence microspectroscopy (μ-XRF). In this study we show that (1) where conventional histochemical procedures revealed only weak deposits of iron and calcium, μ-XRF analysis is able to detect significant deposits of both iron and calcium on the inhaled asbestos fibers; (2) the extent of the deposition of these ions is proportionally directly related and (3) iron and calcium deposition on inhaled asbestos fibers is concomitant with the appearance of inflammatory and hyperplastic reactions.

The time of death can be established by determining the length of the postmortem interval. Many methods have been proposed to achieve this goal. Flow cytometric evaluation of DNA degradation seems to be reliable for the first 72 hours after death. Our study evaluated the correspondence of the corruption process between in vitro and corpse tissues. We chose spleen tissue to perform our investigation because it is rich in nucleated cells. Results showed a precise correspondence between the two kinds of samples in the time period between 24 and 36 hours. The period from 36 to 72 hours is characterized by a much looser correspondence than that found in the first period. After the first 72 hours, DNA denaturation is massive and does not allow useful cytofluorimetric readings. The spleen does not seem to be the most suitable organ for this type of investigation because it tends to colliquate very rapidly. We therefore are evaluating other organs to identify a more suitable tissue source for the investigation of longer postmortem period using flow cytometry.

Environmental and occupational inhalants may induce a large number of pulmonary diseases, with asbestos exposure being the most risky. The mechanisms are clearly related to chemical composition and physical and surface properties of materials. A combination of X-ray fluorescence (μXRF) and Fourier Transform InfraRed (μFTIR) microscopy was used to chemically characterize and compare asbestos bodies versus environmental particulates (anthracosis) in lung tissues from asbestos exposed and control patients. μXRF analyses revealed heterogeneously aggregated particles in the anthracotic structures, containing mainly Si, K, Al and Fe. Both asbestos and particulates alter lung iron homeostasis, with a more marked effect in asbestos exposure. μFTIR analyses revealed abundant proteins on asbestos bodies but not on anthracotic particles. Most importantly, the analyses demonstrated that the asbestos coating proteins contain high levels of β-sheet structures. The occurrence of conformational changes in the proteic component of the asbestos coating provides new insights into long-term asbestos effects.

Ovarian-type mucinous tumors may occasionally occur in the retroperitoneum, pancreas, and liver exclusively or almost exclusively in women. In men, only few cases of such neoplasms arising within or around the testis have been reported. We describe a unique case of an ovarian-type mucinous cystadenoma occurring in the peritoneal cavity of a 65-year-old male patient with secondary adrenocortical insufficiency and hypogonadism. There was a typical fallopian tube enclosed in the capsule of the tumor. Contiguous to that, accessory ectopic epididymal tissue was found. We have interpreted this case as the result of a minor disorder of embryonic development involving structures of both müllerian and wolffian origin. The disproportion of sexual hormones might have had a role in inducing intra-abdominal müllerian remnants to give rise to the mucinous cystadenoma.

The aim of our study was to follow-up 15 patients with stretch marks treated positively with the CuBr laser (577-511 nm) in 1998-99 and followed-up for 2 years.The patients were Italian women, young to middle age (average 30 years old), with skin coloration classified as Fitzpatrick II-III. Biopsies were taken on some patients before the treatment and 1 month after the first treatment. Double-blind histological, histochemical and photographic evaluation was performed. Results obtained as well as to the contradictory effects reported elsewhere in the literature were compared.On average, the results were positive and there were some pathogenic considerations that justified the use of laser.

Depth of invasion and stage of the disease are well known prognostic indicators in cutaneous melanoma (CM). However, the role of other parameters, such as the variations in cytomorphology of melanocyte tumours, mitotic activity and angiogenesis is still open to question. The aim of this study was to analyse proliferation by mitotic activity index (MAI) and immunostaining of proliferating cell nuclear antigen (PCNA), and the intensity of neovascularization (microvessel density; MVD) in CM clinical stage I in relation to epithelioid, spindle and nevoid cell type, histological type (superficial spreading melanoma and nodular melanoma), Clark's level and Breslow thickness. Finally, the role of all parameters in the prognosis of CM was evaluated. Statistical analysis demonstrated that cytological characteristics of CM correlate only with Clark's level, while histological types correlate with MAI, PCNA and MVD. MAI and PCNA also showed correlation between groups according to Clark's level and Breslow thickness. Finally, tumour cell PCNA was found to correlate with MVD. Survival of patients with CM correlated significantly with MAI. These results suggest that cytological variation, histological type, PCNA and MVD alone are not independent prognostic parameters, whereas MAI is a potentially important prognostic marker in CM.

Aims —The spectrum of microscopic lesions resulting from the chronic use of non-steroidal anti-inflammatory drugs (NSAIDs), known as chemical gastritis, remains unclear, and the variable prevalence reported in different studies makes this issue a matter of lively debate. The aim of this study was to evaluate the prevalence and importance of chemical gastritis in patients regularly taking NSAIDs. Owing to the high prevalence of Helicobacter pylori infection, particularly in subjects over 60 years of age, and in view of a possible association with damage, the presence of H pylori infection in the same tissue sample was also determined in all patients. Methods —One hundred and ninety seven subjects were enrolled, 118 of whom were receiving chronic treatment with NSAIDs and 79 of whom were controls, pair matched for age, sex, and clinical symptoms (ulcer-like dyspepsia or upper digestive tract haemorrhage). Antral biopsies taken during upper gastroduodenal endoscopy were assessed for chemical gastritis according to a modified version of Dixon9s score, and for helicobacter correlated chronic active gastritis, according to the updated Sydney system. Results —Chemical gastritis was identified in 11 patients taking NSAIDs (9%) and in four controls (5%) (p Helicobacter pylori was detected in 53 patients taking NSAIDs (45%) and in 34 controls (43%). Patients taking NSAIDs had a significantly higher number of erosions and ulcers and worse endoscores than controls. The presence of H pylori did not appear to increase histological damage, ulcer prevalence, or haemorrhagic events. Conclusions —Chemical gastritis is present in a limited number of patients regularly taking NSAIDs, and is not strongly correlated with NSAID induced damage. In many cases of peptic ulcer or upper gastrointestinal bleeding in patients taking NSAIDs, the presence of chemical gastritis or H pylori infection cannot solely account for the development of mucosal damage.

The oral complications of diabetes mellitus are generally considered to be no more than inevitable annoyances and, as such, have been the subject of relatively few scientific and clinical studies. This review highlights both the known correlations between oral disease and diabetes mellitus, such as those involving supporting tissues and oral mucosa, and those that are still a matter of debate, such as dental caries. In all cases, we carefully analyzed the pathogenesis of the disorder with a view to establishing a possible therapeutic approach, an aspect that currently appears to be overlooked. In view of the fact that the treatment of certain oral conditions, in particular periodontal disease, has been reported to improve metabolic control in diabetic patients, we conclude that diabetes mellitus-related oral diseases deserve adequate recognition and further investigation.

The study of the biologic processes involved in determining the postmortem interval also can be useful for determining the time of death. In this instance, we investigated the time of death in a case of homicide-suicide using circumstantial findings and thanatochronological and flow cytometric data. The flow cytometric data confirmed the time of death as calculated from the police investigations. This method has proven more reliable than thanatochronological techniques and can be used to supplement these observations. This case indicates that the use of flow cytometry "on the spot" is useful and reliable; however, more evidence is needed to support this new, although to date very sound, method of investigation. Language: en

The authors tested the Soluene-350 method to detect diatoms in three cases of sea water drowning. The negative results obtained in all three cases prompted us to re-test the method on samples of fresh water and sea water microalgae. The experiment showed that the siliceous frustule of sea water diatoms is solubilized by Soluene-350 while that of fresh water diatoms is resistant to the treatment. The method, which is effective in cases of drowning in fresh water, should therefore be used with caution in sea water drowning.

In cases of rape, obtaining enough biologic material for DNA identification of the attacker is often difficult because the methods for distinguishing and separating sperm cells from vaginal cells are not sufficiently efficacious. This article describes a new, innovative method for spermatic DNA extraction from the vaginal washing fluid by means of flow cytometry. The high specificity and sensitivity of the flow-cytometric sorting method provides enough sperm cells for DNA typing. The ease of execution of this method, involving vaginal washing with physiologic solution and flow-cytometric reading of the fresh sample, substantially increases its cost-benefit ratio.

In this study, we have performed a morphological analysis of crocidolite fibres interaction with mesothelial cells (MET5A) by combining conventional electron microscopy with atomic force (AFM) and scanning near-field optical microscopy (SNOM). After 6-h exposure at a crocidolite dose of 5 μg cm(-2), 90% of MET5A cells interact with fibres that under these conditions have a low cytotoxic effect. SEM images point out that fibres can be either engulfed by the cells that lose their typical morphology or they can accumulate over or partially inside the cells, which preserve their typical spread morphology. By using AFM we are able to directly visualize the entry-site of nanometric-sized fibres at the plasma membrane of the spread mesothelial cells. More importantly, the crocidolite fibres that are observed to penetrate the plasma membrane in SNOM topography can be simultaneously followed beneath the cell surface in the SNOM optical images. The analysis of SNOM data demonstrates the entrance of crocidolite fibres in proximity of nuclear compartment, as observed also in the TEM images. Our findings indicate that the combination of conventional electron microscopy with novel nanoscopic techniques can be considered a promising approach to achieve a comprehensive morphological description of the interaction between asbestos fibres and mesothelial cells that represents the early event in fibre pathogenesis.

International audience; Asbestos is a potent carcinogen associated with malignant mesothelioma and lung cancer but its carcinogenic mechanisms are still poorly understood. Asbestos toxicity is ascribed to its particular physico-chemical characteristics, and one of them is the presence of and ability to adsorb iron, which may cause an alteration of iron homeostasis in the tissue. This observational study reports a combination of advanced synchrotron-based X-ray imaging and micro-spectroscopic methods that provide correlative morphological and chemical information for shedding light on iron mobilization features during asbestos permanence in lung tissue. The results show that the processes responsible for the unusual distribution of iron at different stages of interaction with the fibres also involve calcium, phosphorus and magnesium. It has been confirmed that the dominant iron form present in asbestos bodies is ferritin, while the concurrent presence of haematite suggests alteration of iron chemistry during asbestos body permanence

The level of degradation of DNA as a means for determining the time of death has been proposed as a valid adjunct to the classic thanatochronologic methods. The twofold aim of this work was to determine which organ might reveal both a correlation between the percentage of degradation of the DNA and the time lapse since death, and would be easiest to sample and yield the most reproducible results even in technically unfavorable situations such as on-the-spot investigations at the scene of death. A comparison of the spleen, blood, and liver showed that hepatic tissue best meets these specific needs because it shows a virtually linear correlation between the time elapsed since death and the level of degradation of the DNA, and it can easily be sampled at the scene of death by use of a common biopsy needle.

Gastrointestinal stromal tumors (GISTs) are the most common primary mesenchymal tumors of the gastrointestinal tract, and most of them harbor KIT or platelet-derived growth factor receptor alpha (PDGFRA) gain-of-function mutations. Proper diagnostic assessment of GISTs has become very important since the availability of the molecular-targeted therapy with imatinib mesylate. Histopathology remains the gold standard in GIST diagnosis, and immunohistochemistry plays the major confirmatory role. Moreover, genetic sequencing not only further confirms the diagnosis of GIST, but also provides information for the optimal treatment of patients. Herein, we describe a gastric GIST harboring a novel PDGFRA exon 14 frameshift mutation caused by a single-nucleotide deletion. The case reported here represents further evidence regarding the existence of a distinct subset of GISTs characterized by the PDGFRA mutation, the gastric localisation, the epithelioid morphology, and the weak or negative immunohistochemical expression of KIT. DOG1 is emerging as a promising biomarker for this subgroup of GISTs.

Ferruginous bodies (FB) are polymorphic structures whose formation is macrophage dependent, and are composed of a core, which may consist of an asbestos fiber coated with proteins, among which ferritin is the main component. Within ferritin, the ferric and ferrous ions are coordinated as ferrihydrite, which is the main iron (Fe) storage compound. However, when ferritin accumulates in some tissues following Fe overload it also contains magnetite along with ferrihydrite, which endows it with magnetic properties. Recently studies showed that magnetite exerts peroxidase-like activity, and since ferruginous bodies display magnetic properties, it was postulated that these particular structures may also contain magnetite within the ferritin coating, and thus may also exert peroxidase-like activity. Histochemical analysis for peroxidase of isolated FB smears demonstrated positive staining. Samples isolated from 4 different autopsy lung fragments were also able to oxidize 3,3',5,5'-tetramethyl-benzidine to a blue colored compound that absorbs at 655 nm. This activity was (1) azide and heat insensitive with optimal pH from 5 to 6, and (2) highly variable, changing more than 25-fold from one sample to another. These findings, together with evidence that the peroxidase-like activity of ferruginous bodies has a hydrogen peroxide and substrate requirement different from that of human myeloperoxidase, can exclude that this enzyme gives a significant contribution to the formation of FB. Standard Fe-rich asbestos fibers also express a peroxidase-like activity, but this appears negligible compared to that of ferruginous bodies. Strong acidification of standard Fe-containing asbestos fibers or magnetically isolated ferruginous bodies liberates a high amount of peroxidase-like activity, which is probably accounted for by the release of Fe ions. Further, FB also damage mesothelial cells in vitro. Data suggest that FB exert peroxidase-like activity and cytotoxic activity against mesothelial cells, and hence may be an important factor in pathogenesis of asbestos-related diseases.

Background. Defects in the mitotic spindle checkpoint have been proposed to contribute to the chromosomal instability observed in human cancers, including oral squamous cell carcinoma (OSCC). BUBR1 is a key component of the spindle checkpoint, whose role in oral carcinogenesis still needs to be clarified. Methods. We have analyzed the expression of BUBR1 in 49 cases of OSCC by immunohistochemistry and compared the findings with clinicopathologic parameters, proliferative activity, and DNA ploidy. Results. BUBR1 was overexpressed in 11 cases (22.4%). Tumors with overexpression of BUBR1 were associated with a less advanced pathologic stage (p = .05) and showed longer survival periods (p = .38) but shorter recurrence-free survival periods (p = .13) than those without it. Conclusions. Our data imply the possibility that BUBR1 may be involved in the progression of OSCC, and suggest that BUBR1 may be a promising prognostic marker in patients with OSCC. © 2010 Wiley Periodicals, Inc. Head Neck, 2011

We present a case of primary pericardial mesothelioma occurring in an asbestos-exposed 67-year-old man who underwent four aortocoronary bypass grafting seven years prior to the onset of the mesothelioma. Primary pericardial mesothelioma is a rare tumor whose association with asbestos is more infrequent than that of the much more common pleural form. Factors other than asbestos that may play a role include genetic predisposition, immune impairment, infections, radiation, dietary factors, and recurrent serosal inflammation. We consider that, in the presented case, inflammation and healing resulting from pericardiotomy might have had a synergistic effect with asbestos in the pathogenesis of the tumor. To our knowledge, this is the first reported case of primary pericardial mesothelioma arising in a patient exposed to asbestos who previously underwent cardiac surgery.

Objectives: Stem cells have widely shown to be able to calcificate both in vitro and in vivo when implanted under the skin and if conveniently differentiated. However nowadays, in scientific literature a study on their efficiency in the osseous regeneration does not exist yet, thus this aspect was investigated in this work. Materials and methods: 10 White Rabbit New Zealand were submitted to a first surgical procedure, which consisted in a sampling of interscapular adipose tissue, from which some adult stem cells were isolated, expanded and differentiated towards the osteogenic line. In a second surgical procedure 5 different defects on esch animal’s calvarium were created, which had a 0.6cm diameter and were 0.2cm deep (this measures were measured by an appropriate caliper). 4 defects were regenerated using 2 different custom-made scaffolds produced by Geistlich®, with and without the aid of stem cells. After the insertion, the site was covered with a collagen membrane fixed by 5 modified titan pins. The defect in the front was left empty on purpose as an internal control for each animal. These animals were sacrificed in pairs, respectively after 2, 4, 6, 8, 10 weeks. The samples were analyzed by MicroCT analysis using a synchrotron light, through the attainment of 1200 slices for each sample. Once slices were binarizated through an Otsu algorithm, stereologic parameters were calculated and, after that, each sample was submitted to histologic analysis. Results: Histological analysis: Regeneration occurred in an optimal way in each sample treated with scaffolds. The use of adult stem cells combined with scaffolds, seemed to accelerate some steps of the normal osseous regeneration: in fact we assisted to a precocious new deposition and calcification of the extra-cellular matrix and to a colonisation of the osteocyte-lacking areas present in the bovine bone. Only the non-regenerated defect showed a deficit in the calcification of the sample up to the 10th week. MicroCT analysis: A similar behaviour for all the analyzed samples was observed: the quantity of new bone (higher gray levels) showed a tendency to increase with the age of the animals. When comparing the histograms obtained for both Bioss and Bioss with stem cells, the quantity of new bone for the Bioss+stem cells samples was higher than for the Bioss alone ones, when age was considered. The same conclusion was obtained by the histological analysis. Conclusions: Further studies on this subject will individuate the best biomaterial to sustain the potential shown by these cells and will quantify their effective contribution to regeneration.

This study, aimed at elucidating the epidemiological features of primary liver carcinoma developing in non-cirrhotic livers, was based on 25 103 autopsies performed between 1975 and 1984 in Trieste, Italy. These autopsies correspond to approximately 70% of all deaths that occurred in this area. Various factors allegedly related to carcinomas were analysed in reference to our previous study on cirrhotic livers and in comparison with 5603 autopsies in Kurume, Japan. There were 28 cases of hepatocellular carcinoma (HCC), 16 of cholangiocellular carcinoma (CCC) not associated with cirrhosis in Trieste, and 48 HCC and 19 CCC in Kurume. On the basis of our findings, it was concluded that cirrhosis, regardless of its cause, is the main pathogenetic factor in HCC; it is responsible for a much higher frequency (14.2:1) than in non-cirrhotic livers, as well as for early occurrence of tumours (an average of 6 years earlier in cirrhotic liver) in Trieste. Patients in Trieste were older than those in Japan, and the frequency of HCC among all autopsies was much greater in the latter. By contrast, the influence of cirrhosis on cholangiocellular carcinoma (CCC) was negligible, as such association appeared purely coincidental or absent. The incidence of CCC among autopsies was greater in Japan. Our data on CCC were not sufficient to demonstrate any clear aetiopathogenetic association between this tumour and alcohol abuse and hepatitis B virus (HBV) infection, except for a possible aetiological role of gallstones. The frequency of CCC relative to HCC was greater in Trieste than in Japan; the incidence of HCC was much less in Trieste, whereas CCC was more frequent in Japan.

The role of asbestos bodies (and associated proteinacious coating) in asbestos associated diseases is not well understood. Currently employed methods of isolation of these bodies employ harsh chemicals that lead to destruction of their proteinacious coating. In this work a method was developed that enabled the purification of whole, integral, unmodified asbestos bodies (AB) by exploiting their magnetic properties. Albumin and ferritin were found to be the major proteins associated with AB isolated from lung tissue of mesothelioma patients. Magnetically isolated AB were shown to be cytotoxic and to activate free radical production from inflammatory cells at a higher extent than that induced by bodies obtained by chemical digestion. The finding that hypochlorite-treated AB induce DNA damage, while AB obtained by the method described in this article failed to do so, together with the differential behavior of these bodies toward inflammatory cells, suggests that native asbestos bodies should be used to investigate the pathogenetic role of these structures.

The case of a 64-year-old man presenting dysuria and haematuria is described. The cause of these symptoms was related to a splenomegaly compressing the left kidney and renal pelvis. A splenectomy was performed. Macro- and microscopic examination of the mass revealed an inflammatory pseudotumour of the spleen, composed of a variable mixture of polyclonal lymphocytes, eosinophils, neutrophils, plasmacells, foamy hystiocytes, giant cells and fibrous tissue. The aetiology is unknown. An inflammatory pseudotumour of the spleen is clearly a reactive lesion resulting from a variety of causes, particularly vascular thrombosis, infections or autoimmune mechanisms. Splenectomy is both diagnostic and curative.

The aim of this study was to verify the effectiveness and safety of the 808-nm wavelength for use on benign lesions of the oral cavity that are stained with toluidine blue solution.The toluidine blue solution according to Mashberg is used in stomatology for the detection of leuko-erythroplasic lesions. In our experience, we have used it to stain even serous cysts, angiomas, and fibromas. We chose a 808-nm wavelength emitted from a power diode laser, because it is well absorbed by the blue color and barely by the healthy mucosa. The non-contact photocoagulation kept the surrounding borders intact when we used the pulsed-emission, 50 ms 7 W, without anesthesia. No sutures were needed. The leukoplasic borders of the lesions were histologically examined before and after the treatment.The vaporized lesions healed completely in about 5-7 days without any kind of treatment, leaving some slight elastic white scars. A mild edema-erythema followed the treatment for 5-7 days.The selective results confirm the effectiveness of this therapeutic procedure on pigmented benign lesions, pre-treated with exogenous chromophores and photocoagulated without bleeding, with a minimum energy 808-nm laser.

The aim of this work was to characterize p53 and PCNA expression in primary tumours and lymph node metastases of oral/oropharynx carcinomas by flow cytometry and to evaluate the possibility of a relationship between these parameters in primary tumours and locoregional metastasis, clinical-histopathological parameters and survival time. A group of 28 patients was retrospectively selected: 13 with metastasis and 15 without metastasis. The following clinical and histopathological parameters were considered: degree of keratinization, Broders' grading of histological differentiation, invasive cell grading, thickness and tumour size. The nuclei of appropriately prepared cells were stained using anti-p53 and anti-PCNA FITC conjugated monoclonal antibodies. Analyses were performed by a Coulter Epics Elite flow cytometer. The following results were obtained: a close relationship between p53 and PCNA in primary tumours but no correlation between them and clinical-histopathological parameters; no correlation between p53 and PCNA expression in primary tumours and in lymph node metastasis; no correlation between p53 and PCNA values in primary tumours and survival. In conclusion, the difference in expression of these biological parameters in primary tumours and in lymph node metastases supports the theory that tumour cells in lymph nodes can display an independent behaviour. Since these biological parameters fail to correlate either with clinical-histopathological parameters or survival, their prognostic role is questionable.

We report on a case of a peculiar tumor of the pilosebaceous unit showing a composite histologic appearance. The case presented as a pigmented crusted lesion on the back of the nose of a 62-year-old woman with markedly sun-damaged skin. Histologically, the superficial portion of the neoplasm was composed of buds and nests of basaloid epithelium with varying degrees of pilar and sebaceous differentiation. Adjacent to this component, lobules of squamous cells with cytoplasmic glycogenation suggesting the mature outer root sheath were seen. In the underlying dermis, there was a well-defined nodular proliferation composed of variably pigmented basaloid matrical cells forming clusters of "shadow" or "ghost cells" admixed with numerous melanized dendritic melanocytes; this last component of the neoplasm was identical to a recently described entity, melanocytic matricoma. The small size, circumscription, and absence of necrosis favored benignity, although the cytologic atypia of matrical cells did not exclude malignancy. The case is interesting not only because it is the third reported case of a peculiar neoplasm imitating the epithelial-melanocytic interaction in the embryonal hair follicle or bulb of the anagen follicle but because the part resembling melanocytic matricoma presented as a component of a complex lesion. We believe that sunlight may have played a role in the development of this peculiar neoplasm.

Gastrointestinal stromal tumors (GIST) are rare neoplasms of unknown etiology and pathogenesis. Their clinical behavior is very unpredictable and a reliable prognostic factor is lacking. The aim of this study was to analyze some prognostic factors and estimate which one is the most reliable. Thirty-eight biopsy specimens of GIST were immunolabeled for PCNA, CD34, vimentin, NSE and actin. The greatest diameter, histological grading, mitotic count, DNA-index and S-phase were estimated for each case. All patients were followed-up for at least 24 months or to death. The data were analysed by univariate and multivariate statistical analysis using a computer program. The results showed that greatest diameter, tumor grade, mitotic count and PCNA-index are prognostic factors in univariate analysis. In multivariate analysis only the greatest diameter is a useful prognostic factor for planning further therapy.

Tumour cell proliferation is an important biological prognostic parameter to be considered alongside clinical and histopathological parameters. It has been evaluated by immunohistochemistry using proliferative markers (PCNA, Ki67, etc.) and by flow cytometry considering DNA content, growth fraction (S + G 2 M) and S-phase fraction. Our aim was to evaluate by flow cytometry both Ki67 and S-phase fraction, S + G 2 M, DNA content in patients surgically treated for oral cavity and/or oropharynx carcinomas. The study, performed on archival material, analysed the significance and prognostic reliability of these biological parameters to reveal locoregional metastasis and evaluated their possible correlation with clinical and histopathological parameters. In conclusion, tumour cell proliferation seems not to be useful in revealing the presence of locoregional metastasis in oral cavity and oropharynx carcinomas. Thus, new approaches are required.

Angiogenic and anti-angiogenic factors, secreted by tumor, inflammatory, and stromal cells play an important role in regulation of neovascularization. Among the most important of these is vascular endothelial growth factor (VEGF), a specific mitogen for endothelium, which increases vascular permeability and induces proteolytic enzymes necessary for vascular remodeling. Tumor-associated macrophages (TAMs) can express complex functions related to tumor biology, including growth, proliferative rate, stroma formation and dissolution, and neovascularization. The aim of this study was to define, using immunohistochemical analysis, the microvessel density (MVD), VEGF expression, and TAMs level in 97 human invasive ductal breast carcinomas not otherwise specified (NOS), investigate a possible relationship between them and then correlate their values with tumor grade, mitotic activity index (MAI), tumor size and lymph-node status. Statistical analysis showed a strong positive relationship between MVD and VEGF expression ( P

This report describes a 25-year-old man who died due to extensive cystic medionecrosis of the aortic wall complicated by both an acute and an ancient dissection. The very unusual finding of an intravascular papillary endothelial hyperplasia, also known as Masson's pseudoangiosarcoma, in the dissected wall is reported given its resemblance to a hemangiosarcoma.

Cutaneous pseudolymphomas refer to a group of lymphoproliferative disorders of difficult classification and diagnosis. Considered as very low-grade lymphomas by some authors, they are actually able to progress to overt cutaneous lymphomas in a minority of cases. We report a case of cutaneous pseudolymphoma of the breast in a 21-year old woman which relapsed locally ten years later. The case is of interest for the unusual localisation of the lesion, for its recurrence after such a long period of time, and for the homozonal site of recurrence. The reported features support both the long course and the lymphomatous nature of such lymphocytic proliferations.

Lasers with a wavelength between 351-1064nm, preset fluence and pulse duration induce a selective thermolysis of tissue chromophores. Specific exogenous chromophores, such as indocyanine green, are used in ophthalmology, in photodynamic therapy, in welding, and more recently for epilation. The aim of our study was to assess the effectiveness of a wavelength of 808nm on tissues stained with different exogenous chromophores whose absorption curves we studied by spectrophotometer. Out of five chromophores, methylene blue proved to be effective at 808nm. We tested the diode laser 808nm, at focused powers of 1, 2, 3, 5, 7 W in cw, for 1, 2, 3, 5, 7, 10 sec on skin specimens stained with 1 percent methylene blue in a small- spot sequence. At low powers the destructive effect was highly concentrated, both in depth and laterally, while greater intensities result in marked destruction even after a short irradiation time. The use of exogeneous chromophores increases absorption and reduces laser energy transmission and scatter. This study confirms that the selectivity of 808nm lasers can be 'tricked' since thermolysis is confined to the stained areas and there is less risk of damaging surrounding areas in the event of abrupt movements by either the surgeon or the patient, especially in the case of emotionally unstable or very young patients.© (2000) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Among malignant neoplasms of the pancreas, lymphomas have a very low incidence. Nevertheless, the dramatic difference in prognosis and treatment between pancreatic carcinoma and lymphoma stresses the importance of a correct diagnosis, especially because most lymphomas of the biliopancreatic region are low-grade B-cell type. The case observed by us shares clinical and pathological features with the few previously reported in literature, and focuses clinicians' attention on the diagnosis of this unusual neoplasm whose clinical behaviour is characterized by obstructive jaundice and overlaps with that of epithelial tumours.

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