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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.

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., (Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Nanotoxicology and nanomedicine investigations often require the probing of nano-objects such as fibres and particles in biological samples and cells, whilst internalization and intracellular destiny are the main issues for in vitro cellular studies. Various high resolution microscopy techniques are well suited for providing this highly sought-after information. However, sample preparation, nanomaterial composition and sectioning challenges make it often difficult to establish whether the fibres or particles have been internalized or they are simply overlaying or underlying the biological matter. In this paper we suggest a novel suitable combination of two different microscopic techniques to reveal in intact cells the uptake of asbestos fibres by mesothelial cells. After exposure to asbestos fibres and fixation, cells were first analysed under the AFM instrument and then imaged under the TwinMic soft X-ray microscope at Elettra Sincrotrone. The suggested approach combines standard soft X-ray microscopy imaging and AFM microscopy, with a common non-invasive sample preparation protocol which drastically reduces the experimental uncertainty and provides a quick and definitive answer to the nanoparticle cellular and tissue uptake.

Ethanol is an essential chemical reagent in DNA preparation as its use increases the yield of extraction. All methodologies for DNA isolation involve the use of ethanol in order to prevent DNA dissolution in water and to optimize the binding of DNA to chromatographic membranes. In this note, we show how the presence of ethanol traces in DNA aqueous solution affects ultraviolet Raman spectra, leading to possible misinterpretations. We report a simple method to remove the ethanol Raman features from the spectra, based on heating the DNA sample at 80 ℃, followed by a slow cooling procedure.

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.

Many drugs, chemicals, and environmental factors can impair sperm functionality by inducing DNA damage, one of the important causes of reduced fertility potential. The use of vibrational spectromicroscopy represents a promising approach for monitoring DNA integrity in sperm, although some limitations exist, depending from the experimental conditions. Here, we report that when using FTIR spectromicroscopy to reveal oxidative stress mediated by Fenton's reaction on hydrated sperm samples, DNA damage interpretation is partially compromised by unexpected cell surface precipitates. The precipitates give a broad band in the 1150-1000cm(-1) infrared region, which partially covers one of the signatures of DNA (phosphate stretching bands), and are detected as iron and oxygen containing material when using XRF spectroscopy. On the other hand, the analyses further support the potential of FTIR spectromicroscopy to reveal cellular oxidative damage events such as lipid peroxidation, protein misfolding and aggregations, as well as DNA strain breaks., (Copyright © 2016 Elsevier Inc. All rights reserved.)

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., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

We report on the use of the UV Raman technique to monitor the oxidative damage of deoxynucleotide triphosphates (dATP, dGTP, dCTP and dTTP) and DNA (plasmid vector) solutions. Nucleotide and DNA aqueous solutions were exposed to hydrogen peroxide (H2O2) and iron containing carbon nanotubes (CNTs) to produce Fenton's reaction and induce oxidative damage. UV Raman spectroscopy is shown to be maximally efficient to reveal changes in the nitrogenous bases during the oxidative mechanisms occurring on these molecules. The analysis of Raman spectra, supported by numerical computations, revealed that the Fenton's reaction causes an oxidation of the nitrogenous bases in dATP, dGTP and dCTP solutions leading to the production of 2-hydroxyadenine, 8-hydroxyguanine and 5-hydroxycytosine. No thymine change was revealed in the dTTP solution under the same conditions. Compared to single nucleotide solutions, plasmid DNA oxidation has resulted in more radical damage that causes the breaking of the adenine and guanine aromatic rings. Our study demonstrates the advantage of using UV Raman spectroscopy for rapidly monitoring the oxidation changes in DNA aqueous solutions that can be assigned to specific nitrogenous bases.

Familial adenomatous polyposis (FAP) is an autosomal dominant inherited syndrome, caused by germline mutations in the adenomatous polyposis coli (APC) suppressor gene. Patients with colorectal polyps are more likely to develop a malignant condition with poor prognosis. Typical FAP is characterized by hundreds to thousands of colorectal adenomatous polyps and by several extra-colonic manifestations; an attenuated form of polyposis (AFAP), presenting less than 100 adenomas and later onset, has been reported. In this study we have examined five Sicilian families affected by FAP syndrome, in order to provide predictive genetic testing for the affected families, as well as to contribute to mutation catalog enrichment. We have detected different APC mutations in these five pedigrees, confirming the remarkable heterogeneity of the mutational spectrum in FAP., (Copyright © 2014 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

HNPCC is an autosomal inherited cancer syndrome characterized by germinal and somatic mutations of DNA mismatch repair (MMR) genes. The inherited mutation in one allele together with an acquired defect in the other allele of an MMR gene leads to accelerate tumor progression. In this study we analyzed a cohort of 11 subjects belonging to four Sicilian families with HNPCC suspected by molecular analysis of coding regions of hMSH2 (NC_000002) and hMLH1 (NC_000003) genes. Molecular analysis has detected the presence of two mutations in gene MSH2 and one mutation in MHL1 gene. In addition, we found a novel mutation consisting in a G deletion at 914 codon of the exon 16 in the MSH2 gene. This deletion leads to a stop codon due to a frame-shift, resulting in a truncated protein. We extended genetic analysis to the other family members and the same mutation was detected in three sisters and in one of the two healthy daughters. This mutation is correlated with clinical findings revealed in genealogic tree and it represents a novel mutation responsible of HNPCC., (Copyright © 2014 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

Breast cancer is a rare, but frequently hidden pathology. A woman, 36 years old, during the early months of pregnancy found a little tumor in her right breast. A fine needle biopsy was negative for cancer. Despite this, the tumor rose and two months after delivery (the patient breast-fed her daughter for a month), she had pain in the right axillary region and the tumors involved all superior dials of the right breast. A Madden mastectomy was performed. The histopathological report was: ductal invasive breast cancer 3 of 19 lymph nodes involved, stage IIIA, TNM pT3N2M0, ER -, PgR +--. Chemotherapeutic regimens were: at first ADM 75 mg/m2 for 5 cycles, and after CMF 1-8 for 6 cycles. After six months the woman had a cutaneous recurrence in the scar of mastectomy, treated with surgery and RT. Thirteen months after, she had lung MTS and then brain MTS. The patient died thirty months after the mastectomy. The surgeons have to discover the women high-risk for the breast cancer before and during the pregnancy. Excisional biopsy is the diagnostic procedure of choice for breast lump during pregnancy. When a breast cancer develops during a pregnancy, the surgeon has to operate immediately the tumors. Chemotherapeutic regimens should be delayed until the second o third trimester or after delivery.

Carcinoma of the main biliary way is a condition that often leads to a fatal outcome owing to the rapid growth and late onset of the symptoms. The importance of a correct, timely diagnosis is stressed, with reference to a clinical case in which even CT scanning did not confirm either the presence of the neoformation or the dilation of the intra and extrahepatic bile ways. The usefulness of humoral markers for the diagnosis of the disease in its preclinical phase in the interests of surgical treatment that is as radical as possible is underlined.

Mucocele is a rare pathology which is difficult to diagnose prior to surgery. The clinical symptoms are aspecific. Diagnosis is generally made at the time of surgery. Explorative laparoscopy is not advised owing to the possibility of encouraging metastatic diffusion. Surgery associated with ex tempore freezer tests must be carried out with the greatest caution to avoid rupture of organs and the spread of mucin. Appendectomy and removal of the adnexa is recommended in benign forms, with or without omentectomy. Omentectomy is necessary together with hystero-adnexectomy and right hemicolectomy in malignant forms, as well as lymph node cleaning and removal of all mucin in pseudomyxoma peritonei.

Giant esophageal polyp is a very rare neoplasm, usually benign but often demanding both for diagnostic and/or therapeutical procedures and for dramatic symptomatologic onset in the patient (regurgitation and asphyxia). The authors present a brief clinical report of a patient with a 20 cm.-long esophageal polyp removed by left lateral cervicotomy and esophagotomy for 8 cm. below the upper esophageal sphincter (histologic examinations showed a pedicled polypoid fibrolipoma with a remarkable presence of myxoid and vascular components). The authors also describe diagnostic and therapeutic methods to manage this rare disease and they gave a brief review of recent literature.

The incidence of Cutaneous Melanoma is 4-5% of all the tumors of the skin. This incidence increases several folds in the last years. Metastases of Melanoma involve lungs, skin, soft tissue, liver, bone, brain, but in 20-30% of the patients involve gastrointestinal (GI) tract. In 60-70% of the cases GI metastases involve small bowel, in 15-20% stomach, in 10-20% large bowel, in 5% esophagus. In 8% of the patients the primary cutaneous melanoma is not known. The prognosis of the patients with metastatic melanoma is poor with an average survival of 5 months. One patient male, 51 years old, underwent surgery for metastases from melanoma in the lymph nodes of the right axilla and in the gastrointestinal tract (ileum). An ileo-ileo anastomosis and a lymphoadenectomy of the nodes of the right axilla were performed. After a first chemotherapy with DTIC (800 mg/m2) + a-IFN(3MU three times every week) and another with CDDP (30 mg/m2 day 1-3), DTIC (250 mg/m2 day 1-3) and VDS (2.5 mg/m2 day 1) with no response, the patient was treated with chemo-immunotherapy sec. Bernengo, slightly modified: CDDP 75 mg/m2; IL-2 18 MU (9MU b.d.) day 3-6 and 17-21; a-IFN 5MU three times every week. This therapy had a partial response of short-course (three months) and the patient died 15 months after surgery. The authors hope that immunotherapy and genetic therapy improve the survival of the patients with metastatic melanoma in the next years.

The authors present a case of small bowel primitive lymphoma found in a 66-years-old female. After having briefly reviewed the literature they direct their attention to the clinical-instrumental diagnostic difficulties of this neoplasm which is often discovered only when it has already invaded various organs. Therefore surgical therapy is not always radical. This fact in addition to the fatally rapid postoperative course led the authors to discuss the problems involved in a correct treatment of this pathology.

The authors report their experience of 23 patients with ulcerative colitis treated with surgery from 1991 to 1994. At admission 8 patients had a high-grade illness, 10 patients had a middle-grade illness, 5 patients had a toxic megacolon. All the patients were treated with systemic medical management (mesalazine + methylprednisolone) and topical medical management with rectal steroid (methylprednisolone) and rectal mesalazine by enema, and with nutritional support (Total Parenteral Nutrition). The 5 patients with toxic megacolon, after 48 hours of unsuccessful medical management, underwent surgery with deferred urgency; the other 18 patients underwent surgery after one-four months. The authors prefer the ileorectal anastomosis (IRA), since the rectal lesions are more susceptible to topical therapy. A single-stage IRA was performed in 17 patients. A sigmoid resection has been employed in the first stage in the 18th patient with a local development of the disease in the sigma; a following relapse of the lesions required a total colectomy with a low ileorectal anastomosis. A first-stage subtotal colectomy with ileostomy and Hartmann closure of the rectum with low ileorectal anastomosis at a later date was performed in the 5 patients with toxic megacolon. One of these, 18 years old, died after a heart failure. After surgery, as soon as the patients start moving their bowels all of these had again a systemic therapy with mesalazine for a short period and after they had a topical therapy with mesalazine + methylprednisolone by enema, for a long period. The average postoperative period of admission was 16.3 days. The functional results have been encouraging with an average of 1.6 bowel movements daily and an average of 0.4 nocturnal bowel movements. All the patients had a normal anal sphincter function with an acceptable stool frequency. In male patients there no urinary or sexual defects. In one case of these, there was a low grade of dysplasia, revealed by endoscopic biopsy. The authors conclude that today surgical treatment of ulcerative colitis is not well established.

Peritoneal mesothelioma is a rare neoplasm (annual incidence: 1-2 cases per million in the general population) and forms about 10% of all mesotheliomas. The authors report a case of malignant mesothelioma of the peritoneum in a male, 61 years old. After laparotomy the patient was treated with intraperitoneal administration of cisplatin (40 mg/m2 day 1-2-3-29-30-31) and intravenous administration of mitomycin C (10 mg/m2 day 1). The renal toxicity was avoided with the GSH and with prehydration before and after administration of the cisplatin. The bone marrow toxicity was avoided with the subcutaneous administration of G-CSF (300 mg three times each week) and erythropoietin (10.000 U three times a week). After the first cycle, with the reduction of the ascites, the cisplatin was administered intravenously too. After six cycles of chemotherapy (18 months after laparotomy) the patient is alive and he has a performance status of 1 (ECOG/WHO). The chemotherapy with cisplatin and mitomycin C must be preferred to the anthracycline in all the patient with cardiologic involvement. The cisplatin administered by intracavitary route give a quick response with less systemic toxicity. A review of the literature confirms the rarity of this pathology, linked epidemiologically with exposure to asbestos, and the difficulty of the preoperative diagnosis: in fact cytologic assay and ultrasonographic and TC scan always don't permit to discover a mesothelioma. The laparotomy and the laparoscopy are useful in the P.M. for the possibility of the biopsies and the apposition of the catheters for intracavitary therapy. The response of peritoneal mesothelioma to treatment is poor. The median survival after the appearance of the symptom is less than 18 months.

The authors report a case of liposarcoma in the sub-mandibular region, and they describe the histological species and the way of spreading. They suggest a surgical, non-demolitional therapy, that they carried radical if it is related to a strict follow-up. Anatomical and functional results, after demolishing surgical therapy, are sometimes unacceptable if the real benefits in terms of survival are considered. No supporting therapy has been used because of the presumed insensibility of the neoplasm to the chemo-radiotherapeutic treatments. At 2 years from surgical treatment no local relapses, on distance metastasis have been related.

The diagnosis of varicocele in a female patient is difficult clinically. Until recently, only celioscopy and uterine phlebography were helpful. We have developed a new method for diagnosis of the pelvic varicocele using retrograde phlebography of the ovarian and iliac veins. Thirty-three women with pelvic pains and disorders of the menstrual cycle have been studied. In 15 patients, an insufficiency of the left ovarian vein was revealed. In three of the patients, an analogous insufficiency of the right ovarian vein was also recognized, along with chronic bilateral stasis in the pampiniform plexes. An insufficiency of the left iliac vein was present in seven of the patients without ovarian varicocele. The average age of the women without ovarian reflux was lower (22.5 years) than that of women with reflux (36.6 years) (p less than 0.01). In this last group of patients, the number of pregnancies was greater (30 children) than in the group without reflux (two children) (chi-square = 12.75; p less than 0.001). This indicates how parity plays an important role in the determination of the appearance of a varicocele. We conclude that the diagnosis of pelvic varicocele may be made by a new diagnostic method represented by retrograde phlebography of the ovarian and iliac veins. The use of it in patients with chronic pelvic pains and disorders of the menstrual cycle of unknown nature can be done routinely.

The authors report the results obtained from an endoscopic study on 131 gastric resected patients, who in 18.3% showed a pathology due to suture thread. The Authors dwell upon the clinical picture shown by these patients, and also consider the cases performed on urgency for hematemesis and/or melaena, and upon the diagnostical role of endoscopy. Moreover, they report the results obtained through the endoscopic therapy, which in 100% of the cases allowed the removal of the suture thread.