Tumor and peritumoral adipose tissue crosstalk: De-differentiated adipocytes influence spread of colon carcinoma cells.

Colorectal cancer is the second leading cause of cancer and often has a fatal course. There are many studies in the literature that have described a close functional relationship between the tumor mass and surrounding tissue, or tumor stroma, which is affected by the continuous metabolic exchange that occurs at the interface between tumor and tissues in contact with it. There is much evidence that the presence of adipose tissue in stroma plays a fundamental role in modulating the tumor microenvironment and promote tumor development, growth, and angiogenesis due to its endocrine characteristics. In this analysis, we have studied the alterations of adipose tissue surrounding colorectal tumors with MRI and optical imaging in vivo techniques to monitor tumor progression and also performed histological and molecular analysis. We detected differences in the principal adipose markers expressed by adipocytes residing around the rectal colon and observed that peritumoral adipose tissue is exposed to a mesenchymal transition process that leads to the acquisition of a less differentiated phenotype of adipocyte that represents the main cellular type present in tumor stroma. The mesenchymal transition correlated with the acquisition of more aggressive tumor phenotype and could represent a valid target for tumor therapy.

[Perianal Paget's disease: five cases report]. / Malattia di Paget perianale: presentazione di 5 casi.

AIM: Extramammary Paget's disease is a rare neoplastic condition, often associated with a synchronous or metachronous underlying skin or visceral malignancies. The aim of this study was to evaluate retrospectively the results we got in 5 cases of perianal Paget's disease and to revise what literature have reported about this issue. PATIENTS AND METHODS: Five patients with perianal EMPD were consecutively treated in our Division between March 1996 and December 2006. In 3 cases the disease was limited to the epidermidis, in one case with multiple recurrences there was dermal infiltration, and one patient had a low rectal adenocarcinoma with pagetoid phenomenon. The surgical treatment we performed in all patients was a wide perianal excision, followed by reconstruction with cutaneous grafts; the resection of rectal adenocarcinoma was carried out using the transanal approach. In two cases we performed a temporary stoma with the sigmoid colon to help the wound healing. RESULTS: We didn't record any complication neither postoperative nor at long time. Two patients developed a local recurrence, but none of our patients showed distant metastases. Four patients are alive and free from disease and one, who developed a multiple local recurrences, died for heart failure. DISCUSSION: The studies available in literature clearly distinguish between a primary EMPD (intraepidermal/intradermal) and secondary disease which is associated with anorectal adenocarcinomas and is thought to be a pagetoid phenomenon, while few informations can derive from those cases in which the disease is associated with an underlying cutaneous adnexal carcinoma. Disease tend to relapse even after a radical surgery and can have metastatic spread, also in intraepidermal form. CONCLUSIONS: Perianal EMPD is a complex disease, difficult to recognize and the association with synchronous or metachronous malignancies imposes long term clinical and instrumental follow up.

[LaboRisCh: an algorithm for assessment of health risks due to chemicals in research laboratories and similar workplaces]. / LaboRisCh: un algoritmo per la valutazione dei rischi per la salute da agenti chimici nei laboratori di ricerca e negli ambienti di lavoro affini.

BACKGROUND: Chemical risk assessment in research laboratories is complicated by factors such as the large number of agents to be considered, each present in small quantities, and the very short and erratic periods of exposure, all of which make reliable environmental and biological monitoring particularly difficult and at times impossible. In such environments, a preliminary evaluation procedure based on algorithms would be useful to establish the hazard potential of a given situation and to guide the appropriate intervention. OBJECTIVES: The LaboRisCh model was expressly designed to assess the health risk due to chemicals in research laboratories and similar workplaces. METHODS: The model is based on the calculation of the value of a synthetic single risk index for each substance and compound found in a laboratory and, subsequently, of a further synthetic single risk index for the whole laboratory or, where required, a section thereof. This makes LaboRisCh a compromise between need for information, ease of use, and resources required for the assessment. The risk index includes several items, chiefly the physical and chemical properties, intrinsic hazard potential, amount, dilution, and time of exposure to each agent; waste management; possible interactions; presence and efficiency of collective and individual protection devices, and staff training in good laboratory practices. The value of the synthetic single index corresponds to one of three areas: no risk (green), possible risk (yellow), and certain risk (red). CONCLUSIONS: Preliminary data confirm the model. LaboRisCh appears to be a reliable method for chemical risk assessment in research laboratories and similar workplaces.

[Biomarkers for prevention and early diagnosis of malignant pleural mesothelioma]. / Utilizzo di biomarkers nella prevenzione e diagnosi precoce del Mesotelioma Maligno della pleura.

Improved detection methods for diagnosis of asymptomatic malignant pleural mesothelioma (MPM) are essential for an early and reliable detection and treatment of this disease. Thus, focus has been on finding tumour markers in the blood. 94 asbestos-exposed subjects, 22 patients with MM, and 54 healthy subjects were recruited for evaluation of the significance of 8-hydroxy-2'-deoxy-guanosine (80HdG) in white blood cells and plasma concentrations of soluble mesothelin-related peptides (SMRPs), angiogenic factors (PDGFbeta, HGF, bFGF, VEGFbeta), and matrix proteases (MMP2, MMP9, TIMP1, TIMP2) for potential early detection of MM. The area under ROC curves (AUC) indicates that 80HdG levels can discriminate asbestos-exposed subjects from controls but not from MPM patients. Significant AUC values were found for SMRP discriminating asbestos-exposed subjects from MPM patients but not from controls. VEGFbeta can significantly differentiate asbestos-exposed subjects from control and cancer groups. No diagnostic value was observed for MMP2, MMP9, TIMP1, TIMP2. The sensitivity and specificity results of markers were calculated at defined cut-offs. The combination of 80HdG, VEGFbeta and SMRPs best distinguished the individual groups, suggesting a potential indicator of early and advanced MPM cancers. The combination of blood biomarkers and radiographic findings could be used to stratify the risk of mesothelioma in asbestos-exposed populations.

Diagnosis of asbestos-related pleuropolmonary diseases.

A revision of criteria for diagnosis of asbestos-related pathological conditions was performed studying specially asbestosis, pleural plaques and malignant mesothelioma, also taking into account the problems connected with histopathology. As regards the histological diagnosis of asbestosis, it requires the presence of diffuse interstitialfibrosis in a well inflated tissue remote from the site of a tumour or other large lesion, plus the presence of two or more asbestos bodies in a 1 cm2 section. As regards the imaging diagnosis, the HRTC 4-point scale proposed by Paris et al. (2004) has been adopted:--0 images not suggestive of interstitial pneumonia;--1 modest unilateral or bilateral interstitial abnormalities, involving restricted areas if bilateral;--2 interstitial abnormalities of limited extent, but consistent with a diagnosis of asbestosis, i.e. honeycombing, even without other parenchymal changes and even though unilateral, or else any two abnormal findings among thickened interlobular septa, intralobular lines or subpleural curved lines;--3 numerous bilateral changes on several slices involving more than 2/3 of the posterior third of each hemi thorax. Only points 2 and 3 were considered consistent with the diagnosis of lung fibrosis. Such HRCT findings are not specific for asbestosis, changes in the pleural wall such as diffuse plaques and thickenings contribute to the diagnosis of asbestosis. As regards the pleural plaques and asbestos bodies we remark that they are merely exposition markers. We also discussed the problems the pathologist may encounter in diagnosing mesothelioma; in this field the prospects are encouraging as microarray analysis are beginning to identify new molecular markers for mesothelioma.

Deep burn of the hands: Early surgical treatment avoids the need for late revisions?

An eight years experience in the management of deep hand burns is presented. We reviewed 116 cases of patients surgically treated with escarectomy and skin autograft. Seventy eight of them underwent early surgery within the first 4-6 days from injury, while 38 were conservatively treated in the fist stage and only lately surgery was performed (usually after the 14th day). In all patients, we applied the same post operative topic treatment and rehabilitation protocol. Several months after first admission, only 6(7.7%) of early treated patients needed secondary correction for unaesthetic and functionally debilitating scars versus 14(36.8%) of the late treated group. Our review confirm the importance of performing early surgery of deep burns of the hands, whenever possible, in order to achieve best results and shows it's utility in reducing re-admissions for secondary revisions.

Crystalline silica induces apoptosis in human endothelial cells in vitro.

We investigated whether incubation of cultured human aortic endothelial cells (HAEC) with crystalline silica at the concentration 1 cm2/ml (chosen on the basis of a pilot experiment) leads to alterations typical of apoptosis. The binding of annexin V as early, and DNA fragmentation as late events of apoptosis were measured besides the number of cells with depolarized mitochondria. The generation of reactive oxygen species (ROS) by HAEC in presence of silica was determined as well as silica ability to in vitro generate hydroxyl radicals was investigated. After 18 h of silica incubation, about 30% of viable cells bound annexin V. After 24 h of silica treatment, the percentage of cells with fragmented DNA (Tunel positive) was 27% and it increased up to 50% after 48 h, whereas in untreated cells this percentage was 7% and 11% after 24 and 48 h, respectively. The presence of fragmented DNA in cells treated with silica was confirmed by agarose gel electrophoresis. In agreement with these results showing an induction of HAEC apoptosis by silica incubation, the number of cells with depolarized mitochondria was significantly higher after silica treatment as compared to the control. Apoptosis was also obtained with silica added to aliquots of anti-C5a-absorbed-medium. In the cells exposed to silica there was a significant increasing of ROS generation in comparison to the untreated cells. Apoptosis might be due to peroxidative stress since silica can generate hydroxyl radicals.

[Biological response to diverse polymorphic forms of silica]. / Risposte biologiche con diverse forme polimorfe di silice.

Under two different experimental conditions (cleavage of C5 complement fraction and human endothelial cells cultures), crystalline and amorphous silica samples were tested. Different biological reactions were achieved.

[Presence of asbestos in naval vessels: study of airborne fiber concentrations on Italian railways ferry boats]. / Presenza di amianto sulle navi: studio sulla concentrazione di fibre aerodisperse su navi traghetto delle Ferrovie dello Stato.

Asbestos has been widely used on ships as an insulating material due to its fire-resistant and sound-absorbent properties. It was therefore deemed appropriate to measure the concentrations of airborne asbestos fibres on two ferries in regular service on the Civitavecchia-Golfo Aranci route. Decontamination work had been carried out on these vessels in order to remove, as far as possible, or to confine asbestos. Between 1994 and 1998, the Ultrastructures Laboratory of the Istituto Superiore di Sanità analysed 123 samples via scanning electron microscopy (SEM) with elementary dispersion X spectroscopy (EDXS). The results showed that in 98.4% of the samples concentrations in the air were below 2 fibres/litre, which is the value set by Ministerial Decree of 6/9/94 for buildings to qualify for effective decontamination status. Concentrations above this value, between 2 and 10 fibres/litre, were found in one ferry engine room, where workers were not continuously present since a remote control box was installed there. The results show that there is no environmental dispersion of asbestos fibres in usual navigation conditions; the situation is comparable to that in buildings where asbestos-containing safety materials have been used and that are subject to period checks in accordance with the guidelines included in Ministerial Decree of 6/9/94. Legislation is deemed necessary to make law 257/92 applicable to naval vessels in order to define univocal criteria for risk evaluation, checking, maintenance and custody procedures of asbestos-insulated vessels, and procedures for each job on board involving handling or removal of asbestos-containing material.

[Measurement of airborne asbestos fibers on railroad rolling stock]. / Ricerca di fibre di amianto aerodisperse a bordo di rotabili ferroviari circolanti.

In February 1995 the Italian Railways Health Department set up a special study group in order to assess the effectiveness of the measures adopted against hazards due to the presence of asbestos in rolling stock currently in use on the rail network. The group set up specific procedures for sampling and analysis, on the basis of the criteria fixed for civil buildings in Ministerial Decree of 6/9/94, which was subsequently applied to rolling stock by Ministerial Decree of 26/10/95. In accordance with these procedures the study group carried out environmental studies via test runs programmed by the Railways Technical Departments, on trains made up of different types of vehicles. Insulated, completely or partially deinsulated and originally non-insulated vehicles were studied. Samples were analysed via scanning electron microscopy (SEM) with elementary dispersion X spectroscopy (EDXS) carried out by highly qualified public laboratories (ISPESL--National Institute for Prevention and Work Safety, ARPA--Regional Environmental Protection Agency, CRA--Veneto Region, University Departments). Altogether, from the start of the programme up to September 1998, 1464 samples in 170 test runs on 619 rolling stock vehicles were examined. These involved 83 locomotives, 83 electric rail-cars and 453 carriages. The results showed that in over 99% of the samples the fibre concentrations were below 2 fibres/litre, which is the value fixed by law for buildings and rail vehicles in order to qualify for effective decontamination status. Values exceeding 2 fibres/litre were found in only 4 vehicles, which were withdrawn or blocked for further checks. As a precaution, 18 vehicles where concentrations over 1 but less than 2 fibres/litre were found, were also blocked and their return to service has been postponed for further checks and analyses until the results show concentrations below 1 fibre/litre. Environmental analyses carried out up to the present indicate an overall situation comparable to that usually found in the general environmental without any asbestos dispersion sources. Surveillance and investigations are still under way in order to achieve the aims of the safety programme set up by the Italian Railways to ensure health and environmental protection.

In vitro cleavage by asbestos fibers of the fifth component of human complement through free-radical generation and kallikrein activation.

Chrysotile and crocidolite fibers incubated in normal human plasma (NHP) generated from the C5 component of complement C5a-type fragments that stimulated polymorphonuclear leukocyte (PMN) chemotaxis. Absorption of NHP with antiserum against C5a totally abolished neutrophil chemotactic activity. Asbestos fibers also produced C5a small peptides in the presence of ethylene glycol bis(beta-aminoethyl ether) N,N,N'N'-tetraacetic acid (EGTA) but not ethylene diamine tetraacetic acid (EDTA). Activation of C5 was significantly inhibited when asbestos fibers were pretreated with iron chelators such as sodium dithionite (DTN), deferoxamine (DFX), or ascorbate (AA). Concentration-related inhibition of C5 activation was also observed when asbestos fibers were added concurrently to plasma in the presence of DFX, 1,3-dimethyl-2-thiourea (DMTU), a strong hydroxyl scavenger, or aprotinin (APR), a specific protease inhibitor. Further, chrysotile and crocidolite significantly increased plasma kallikrein activity. Data demonstrate that asbestos-induced C5 activation plays a role in inflammatory reactions characteristic of asbestosis through mechanisms involving iron ions, hydroxyl radicals, and oxidized C5-ike fragments. The ferrous ions present at the asbestos fiber surface trigger this activation and catalyze, via Fenton reaction, the production of hydroxyl radicals, which in turn convert native C5 to an oxidized C5-like form. This product is then cleaved by kallikrein, activated by the same asbestos fibers, yielding an oxidized C5a with the same functional properties as C5a.

Role of iron in asbestos-body-induced oxidant radical generation.

Asbestos bodies (AB) were harvested from human lung tissue digests and isolated from uncoated asbestos fibers. Samples containing 1000 AB were added to a reactive solution to investigate the ability of AB to oxidize deoxy-D-ribose and generate reactive oxygen species (ROS) in the presence of ascorbate and hydrogen peroxide as determined by formation of thiobarbituric acid (TBA)-reactive products. Three types of asbestos fibers were tested for comparison, since they are known to be able to produce ROS. The absorbance values measured with 1000 AB were significantly higher than those observed with 1000 fibers of the three types of asbestos. Since in our reaction system the only source of transition metals was the iron-rich AB, data suggest iron derived from the ferritin coating of AB was involved in oxidant generation. Addition of iron to AB enhanced TBA-reactive product formation, while chelation of Fe with deferoxamine reduced this reaction. Hydroxyl radical scavengers 1,3-dimethyl-2-thiourea (DMTU) and mannitol (MN) also effectively blocked TBA-reactive product generation. Data indicate the importance of Fe in AB-induced oxidant damage. With the addition of polymorphonuclear leukocytes (PMN) to AB, incubation in the reactive solution gave very high amounts of TBA-reactive products, but using a reactive solution devoid of ascorbate, very low amounts of TBA-reactive products were generated. In the latter condition, the superoxide of cell membranes probably reduced and removed iron from AB-coating ferritin, but less effectively than ascorbate. Further after the possible reoxidation of Fe2+, Fe3+ could be coordinated by lactoferrin. Since such availability of reductant is never approached in living systems, the iron in the AB coating is unlikely to function as a catalyst of Fenton-type reactions in vivo.

Wollastonite fibers in vitro generate reactive oxygen species able to lyse erythrocytes and activate the complement alternate pathway.

Wollastonite fibers were tested in vitro for their ability to produce reactive oxygen species (ROS) with two different systems: a cell-free reactive mixture containing deoxyribose and a polymorphonuclear leukocyte suspension. After adding the fibers, we measured the thiobarbituric acid-reactive substances produced by deoxyribose degradation and luminol-enhanced chemiluminescence, respectively. Compared with asbestos, wollastonite fibers produced higher ROS levels both in the PMN suspensions and in the cell-free reactive mixtures. A large amount of these ROS were not hydroxyl radicals. Indeed we obtained remarkable differences in ROS generation between unground and ground wollastonite fibers and negative results with fibers modified with ferric chloride and dithionite. In addition, ROS generation was partially inhibited (by 46-54%) in the reactions performed in the presence of 1,3-dimethyl-2-thiourea (DMTU), a strong hydroxyl radical scavenger. Wollastonite fibers were also analyzed for their ability to lyse erythrocytes and activate complement. Hemolytic potency was about twice that of chrysotile and half that of crocidolite. The levels of complement activation (via the alternate pathway) were about four-fifths of those measured in zymosan-activated plasma (a typical stimulus used to activate the alternate pathway), equal to those obtained with crocidolite, and two-thirds of those found with chrysotile. The addition of DMTU markedly reduced both these activities. Since asbestos fiber toxicity is mainly due to hydroxyl radical generation, our results indicate that wollastonite fibers are probably less toxic than asbestos fibers.

Decrease in peripheral blood polymorphonuclear leukocyte chemotactic index in endometriosis: role of prostaglandin E2 release.

OBJECTIVE: To investigate the effect of disease on peripheral blood polymorphonuclear leukocyte chemotactic index and natural killer cell cytotoxicity and to provide additional information concerning the cell-mediated immune function in endometriosis. METHODS: Chemotactic index of peripheral blood polymorphonuclear leukocytes, natural killer cell activity, and plasma estradiol (E2) and plasma prostaglandin (PG) E2 levels were evaluated in 46 women who underwent laparoscopy or laparotomy for pelvic pain, infertility, and/or benign adnexal masses. RESULTS: The 20 women (43%) with endometriosis showed a decrease in peripheral blood polymorphonuclear leukocyte chemotactic index, related to advanced disease stage (P < .001). A significant inverse correlation was observed between plasma PGE2 levels and chemotactic index in stage III and IV endometriosis (r = -.73, P = .004). Similarly, natural cytotoxicity was decreased significantly with respect to the stage of endometriosis (P = .004) and related inversely to plasma PGE2 levels (r = -.74, P = .003). A direct relationship was observed between PGE2 and plasma E2 levels (r = .59, P = .006). CONCLUSION: Advanced endometriosis is associated with decreased peripheral blood polymorphonuclear leukocyte chemotactic index and natural killer cytotoxicity, which may be related to plasma PGE2 and E2 levels.

[Reactive oxygen species produced by the addition of sepiolite and vermiculite (expanded or not) to suspensions of human polymorphonuclear phagocytes and bovine alveolar macrophages]. / Specie reattive dell'ossigeno indotte dall'addizione di sepiolite e vermiculite (espansa e non) a sospensioni di fagociti, polimorfonucleati umani e macrofagi alveolari bovini.

We have studied a sample of commercial sepiolite and two samples of commercial vermiculite, which are clay minerals advised to replace asbestos. We have in vitro tested their abilities to produce reacting oxygen species (ROS) after they have been added to suspension of human polymorphonuclear leukocytes and bovine alveolar macrophages. The behaviour of sepiolite and vermiculite have been compared with those of asbestos fibres given by Unione Internationale contre le Cancer (UICC) and with kaolin and illite. Sepiolite was not able to induce ROS production, while vermiculite was able to induce a relevant ROS generation, even if the values were always lower than that obtained from chrysotile. Kaolin was able to generate a high ROS production.

Biological effects of contaminated silicon carbide particles from a workstation in a plant producing abrasives.

A sample of silicon carbide dust taken in the field from a plant producing abrasives was studied in vitro. The SiC particles (part unmilled and part milled) were able to disturb the structure of erythrocyte membranes and to lead to blood red-cell lysis; they also either interfered with complement and activated the alternate pathway, or interacted with biological media and polymorphonuclear leucocyte membranes, thus eliciting reactive oxygen species production. These in vitro properties were detected both in original large particles and unmilled particles, over 40% of which were of respirable size. The ability of these SiC particles to produce complement activation in vitro lends support to the previous hypothesis, that the radiographic opacities found in two workers employed in the same area of the plant from which the dust tested was taken are due to a reaction by pulmonary interstitial structures to SiC particle inhalation. It is speculated that SiC particles could act like asbestos, the ability of which to activate complement through the alternate pathway is considered to be one of the mechanisms by which the initial asbestotic lesions and subsequent fibrotic inflammatory infiltrates are generated in the lung.

Distally based dorsalis pedis island flap for a distal lateral electric burn of the big toe.

Reconstruction of the small defects in the distal portion of the foot has always represented a difficult problem. A case of a young man with a deep electric burn of the distal lateral side of the big toe successfully treated with a distally based dorsalis pedis island flap based on the first dorsal metatarsal artery (FDMA) is presented. The donor site area of the dorsum of the foot was grafted, and deambulation was reassumed 3 weeks later. Advantages; limits and anatomical consideration regarding the viability of the flap are also discussed.

Fluid replacement in burned patients.

Burn injury involves a large amount of water, electrolytes and proteins loss trough the burn wound. For this reason, to avoid shock, a wide infusion of fluid is necessary in the first hours after trauma. Many reanimation formulas were proposed in the past years, with different composition: saline, colloids, plasma. The authors have studied 40 burned patients admitted in Verona Burn Center within 4 hours after burn, with burns over 30% of the body surface area. Twenty of them were treated with Baxter reanimation formula (ringer lactated saline, RLS) while the others with Monafo hypertonic lactated saline (HLS), modified by Milan Burn Center. The two randomized groups were assessed and compared. In RLS group total fluid volume infused was higher while sodium requirements was lower than in HLS patients, with statistically significative difference (p < 0.01). On the contrary, in HLS group, potassium administered was perhaps twice if compared with the other. Haematocrit, urine output and urine osmolarity were adequate in both the groups, and did not showed statistical differences, such as serum proteins concentration, that was low in all patients, while a significative difference was noted in urine osmolarity (p < 0.01). A metabolic alkalosis was present in HLS patients, while, on the other hand, serum nitrogen was significantly higher (p < 0.05), in the first 48 hours after burn, in RLS group. Patients were assessed for pre-existing diseases too, and data showed that complications were lower in HLS than in RLS group. HLS resuscitation formula guarantees a good electrolytes balance with lower fluid load, reducing tissue oedema and complication rate. Mortality rate was higher in HLS, may be for an higher Roy index in this group.