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ABSTRACT: Products of different origin, time of collection, and activities fall under the general term of colostrum and, therefore, great variability in composition as well as in the concentration of its components has been reported in the literature. In the present study, we describe the standardization of a bovine colostrum derivative and the characterization of its bioactive components. Evaluation of the most representative agents (lactoferrin, transferrin, IL-2, IFN-γ, tumor necrosis factor, IgG, and IgA) showed that a marked decrease in active components occurs after the first few hours. Bovine colostrum was, therefore, collected up to the fifth hour after delivery from Holstein cows, in the presence of preservatives, and immediately frozen. A protocol of centrifugation, filtration, and lyophilization was then applied to pools of colostrum from at least 30 cows to obtain a stable, sterile, standardized product. Preservatives were removed by dialysis. Evaluation of the active biological components of colostrum showed that the final product of colostrums contained significant and reproducible amounts of bioactive factors, including cytokines, immunomodulating factors, growth factors, and immunoglobulins. The final product appeared, therefore, as a sterile, pyrogen-free, standardized derivative of bovine colostrum with a high concentration of bioactive components.
Journal of Dairy Science 01/2013; · 2.56 Impact Factor
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Breast Cancer Research 04/2012; 3:1-1. · 5.33 Impact Factor
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Breast Cancer Research 04/2012; 3:1-1. · 5.33 Impact Factor
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Endoscopy 01/2011; 43 Suppl 2 UCTN:E34-5. · 5.21 Impact Factor
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A Stacchini,
P Carucci,
D Pacchioni,
G Accinelli,
A Demurtas,
S Aliberti,
M Bosco,
M Bruno,
A Balbo Mussetto,
M Rizzetto, G Bussolati,
C De Angelis
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ABSTRACT: Although endoscopic ultrasound combined with fine needle aspiration (EUS-FNA) is rapidly becoming the preferred diagnostic approach for the sampling and diagnosis of gastrointestinal and mediastinal malignancies, there are limited data as to its use in the diagnosis of lymphoproliferative disorders. Therefore, we carried out a retrospective evaluation of the performance of EUS-guided FNA combined with flow cytometry (FC) as a tool to improve overall sensitivity and specificity in the diagnosis of lymphoma.
Of 1560 patients having EUS-guided FNA during the period of the study, a total of 56 patients were evaluated by cytology with FC after EUS-FNA. There was adequate material to perform FC analysis for all but one case.
EUS-FNA-FC gave a diagnosis of lymphoma in 11 cases and of reactive lymphadenopathy in 20. A specific histological type was defined by FC alone in eight cases. The remaining cases were diagnosed later by cytology and cell block sections: 13 carcinomas, nine granulomatous lymphadenopathies and one mediastinal extramedullary haematopoiesis. One case was considered only suspicious for lymphoma on cytology and FC but was not confirmed on molecular analysis and one had insufficient material for FC.
Our results show that a combination of EUS-FNA-FC is a feasible and highly accurate method, which may be used for the diagnosis and subtyping of deep-seated lymphoma, providing a significant improvement to cytomorphology alone both for diagnosis and treatment planning, as long as immunocytochemistry is available for non-lymphoma cases.
Cytopathology 01/2011; 23(1):50-6. · 1.59 Impact Factor
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ABSTRACT: Irregularity in the nuclear shape, with extensive folds and invaginations of the nuclear membrane (NM), remain the basic diagnostic feature of papillary thyroid carcinoma (PTC). The biological reasons for these irregularities are obscure, but evidence has been presented that they might be linked to RET÷PTC gene translocation. In the present study, we have investigated the hypothesis that the NM irregularities in PTC might be linked to alterations in the expression of lamin B receptor (LBR), a component of the inner NM responsible for the distribution of Lamin B and associated chromatin. Fisher AH et al. already reported on the lack of LBR in PTC, a finding in contrast with the observation that a reduced expression of LBR because of gene mutation is responsible for the lack of nuclear segmentation of granulocytes in Pelger-Huët anomaly. In the present study, we confirmed the lack of immunohistochemical staining for LBR in PTC nuclei, in contrast to a positive staining in intestinal epithelium and stromal cells. However, Western blot and RT-PCR analysis demonstrated a strongly positive reaction in PTC extracts, thus proving an expression of LBR higher in PTC cases and cells than in follicular carcinoma cells. In conclusion, our data suggest that LBR is heavily expressed in PTC cells, but an abnormal folding of the protein might explain its lack of immunohistochemical reactivity and be associated with the anomalous folding of the NM.
Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie 01/2010; 51(4):615-20. · 0.52 Impact Factor
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Endoscopy 02/2009; 41 Suppl 2:E6-7. · 5.21 Impact Factor
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ABSTRACT: In situ hybridizaiton (ISH) is a sensitive and specific technique for detecting nucleic acids in intact cells. Visualization of the target sequences by autoradiography or immunohistochemistry allows their precise subcellular localization and quantitation. The application of ISH techniques has contributed to the understanding of the complex replicative cycle of hepatitis B virus. More recently, hepatitis delta and C virus replication has also been studied by this technique. ISH-based assays have finally been used to follow the replication of cytomegalovirus within the transplanted liver. Although ISH is a powerful tool for the molecular biologist, its clinical significance in the diagnosis and prognosis of human I hepatitis virus infections has yet to be fully evaluated.
Liver International 12/2008; 12(4):217 - 226.
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C Marchiò,
M Iravani,
R Natrajan,
M B Lambros,
K Savage,
N Tamber,
K Fenwick,
A Mackay,
R Senetta,
S Di Palma,
F C Schmitt, G Bussolati,
L O Ellis,
A Ashworth,
A Sapino,
J S Reis-Filho
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ABSTRACT: Pure invasive micropapillary carcinoma (MPC) is a special histological type that accounts for 0.7-3% of all breast cancers. MPC has a distinctive growth pattern and a more aggressive clinical behaviour than invasive ductal carcinomas of no special type (IDC-NSTs). To define the molecular characteristics of MPCs, we profiled a series of 12 MPCs and 24 grade and oestrogen receptor (ER)-matched IDC-NSTs using high-resolution microarray comparative genomic hybridization (aCGH). In addition, we generated a tissue microarray containing a series of 24 MPCs and performed immunohistochemical analysis with ER, PR, Ki-67, HER2, CK5/6, CK14, CK17, EGFR, topoisomerase-IIalpha, cyclin D1, caveolin-1, E-cadherin, and beta-catenin antibodies. In situ hybridization probes were employed to evaluate the prevalence of amplification of HER2, TOP2A, EGFR, CCND1, MYC, ESR1, and FGFR1 genes. aCGH analysis demonstrated that MPCs significantly differed from IDC-NSTs at the genomic level. Gains of 1q, 2q, 4p, 6p, 6q23.2-q27, 7p, 7q, 8p, 8q, 9p, 10p, 11q, 12p, 12q, 16p, 17p, 17q, 19p, 20p, 20q, and 21q, and losses of 1p, 2p, 6q11.1-q16.3, 6q21-q22.1, 9p, 11p, 15q, and 19q were more prevalent in MPCs. High-level gains/amplifications of 8p12-p11, 8q12, 8q13, 8q21, 8q23, 8q24, 17q21, 17q23, and 20q13 were significantly associated with MPCs. A comparison between 24 MPCs and a series of 48 grade and ER-matched IDC-NSTs revealed that high cyclin D1 expression, high proliferation rates, and MYC (8q24) amplification were significantly associated with MPCs. Our results demonstrate that MPCs have distinct histological features and molecular genetic profiles supporting the contention that they constitute a distinct pathological entity.
The Journal of Pathology 08/2008; 215(4):398-410. · 6.32 Impact Factor
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C Marchiò,
M Iravani,
R Natrajan,
MB Lambros,
K Savage,
N Tamber,
K Fenwick,
A Mackay,
R Senetta,
S Di Palma,
FC Schmitt, G Bussolati,
IO Ellis,
A Ashworth,
A Sapino,
JS Reis-Filho
[show abstract]
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ABSTRACT: Pure invasive micropapillary carcinoma (MPC) is a special histological type that accounts for 0.7–3% of all breast cancers. MPC has a distinctive growth pattern and a more aggressive clinical behaviour than invasive ductal carcinomas of no special type (IDC-NSTs). To define the molecular characteristics of MPCs, we profiled a series of 12 MPCs and 24 grade and oestrogen receptor (ER)-matched IDC-NSTs using high-resolution microarray comparative genomic hybridization (aCGH). In addition, we generated a tissue microarray containing a series of 24 MPCs and performed immunohistochemical analysis with ER, PR, Ki-67, HER2, CK5/6, CK14, CK17, EGFR, topoisomerase-IIα, cyclin D1, caveolin-1, E-cadherin, and β-catenin antibodies. In situ hybridization probes were employed to evaluate the prevalence of amplification of HER2, TOP2A, EGFR, CCND1, MYC, ESR1, and FGFR1 genes. aCGH analysis demonstrated that MPCs significantly differed from IDC-NSTs at the genomic level. Gains of 1q, 2q, 4p, 6p, 6q23.2–q27, 7p, 7q, 8p, 8q, 9p, 10p, 11q, 12p, 12q, 16p, 17p, 17q, 19p, 20p, 20q, and 21q, and losses of 1p, 2p, 6q11.1–q16.3, 6q21–q22.1, 9p, 11p, 15q, and 19q were more prevalent in MPCs. High-level gains/amplifications of 8p12–p11, 8q12, 8q13, 8q21, 8q23, 8q24, 17q21, 17q23, and 20q13 were significantly associated with MPCs. A comparison between 24 MPCs and a series of 48 grade and ER-matched IDC-NSTs revealed that high cyclin D1 expression, high proliferation rates, and MYC (8q24) amplification were significantly associated with MPCs. Our results demonstrate that MPCs have distinct histological features and molecular genetic profiles supporting the contention that they constitute a distinct pathological entity. Copyright © 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
The Journal of Pathology 04/2008; 215(4):398 - 410. · 6.32 Impact Factor
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A Sapino,
F Montemurro,
C Marchiò,
G Viale,
J Kulka,
M Donadio,
A Bottini,
G Botti,
A P dei Tos,
A Bersiga,
S Di Palma,
M Truini,
G Sanna,
M Aglietta, G Bussolati
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ABSTRACT: Biotin-labeled trastuzumab (BiotHER) can be used to test for HER2 by immunohistochemistry. We previously showed that BiotHER immunoreactivity is highly correlated with HER2 amplification and indicated that it could be associated with better clinical outcome in advanced breast cancer patients receiving trastuzumab.
Tumor specimens and clinical information from 234 patients who received trastuzumab-based treatments were collected from 10 institutions. HER2 amplification and BiotHER immunoreactivity were assessed centrally. The effect of BiotHER positivity on response rate (RR), time to progression and survival were studied by univariate and multivariate analysis in patients presenting HER2-amplified breast cancer. The pathologic reviews of the assays were blinded to patient outcomes.
BiotHER was positive in 109/194 (56%) HER2-amplified breast cancers and in one not amplified tumor. RRs were 74% [95% (confidence interval) CI 64%-81%] and 47% (95% CI 36%-58%) in BiotHER-positive and -negative tumors, respectively (P < 0.001). BiotHER immunoreactivity was independently associated with increased probability of tumor response (odds ratio 3.848; 95% CI 1.952-7.582), with reduced risk of disease progression [hazard ratio (HR) 0.438; 95% CI 0.303-0.633] and with reduced risk of death (HR 0.566; 95% CI 0.368-0.870) by multivariate analysis.
The results support a role for BiotHER testing in better tailoring trastuzumab-based treatments in patients with advanced HER2-amplified breast cancers.
Annals of Oncology 12/2007; 18(12):1963-8. · 6.43 Impact Factor
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ABSTRACT: Biliary tract brush cytology is increasingly being recognized as a favoured method for evaluating abnormalities of the biliary tract. In order to increase the diagnostic accuracy, we devise a new brush processing method finalized to the complete and ideal cytologic examination of the collected material. Small fragments of the mucosa, of inflammatory cell aggregates or of carcinomas are observed and the results are optimally fixed and allow a definitive histological diagnosis.
Minerva medica 09/2007; 98(4):373-8. · 0.90 Impact Factor
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R Rocca,
C De Angelis,
M Daperno,
P Carucci,
N Ravarino,
M Bruno,
L Crocellà,
A Lavagna,
M Fracchia,
D Pacchioni,
G Masoero,
C Rigazio,
E Ercole,
R Sostegni,
M Motta, G Bussolati,
B Torchio,
M Rizzetto,
A Pera
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ABSTRACT: Diagnosis of pancreatic masses is often difficult. Endoscopic ultrasound-fine needle aspiration has been proposed as the best single-step strategy.
To prospectively evaluate feasibility, effectiveness and safety of endoscopic ultrasound-fine needle aspiration of pancreatic masses in a consecutive study of unselected patients.
Two hundred ninety-three patients were enrolled in two referral Hospitals in Northern Italy. All patients were referred either due to the presence of imaging test abnormalities (suspected or evident masses, or features indirectly suggesting the presence of a mass) or due to clinical or biochemical findings suggesting pancreatic cancer in the absence of positive imaging. All patients underwent linear array endoscopic ultrasound and, when indicated, fine needle aspiration. All procedures were recorded prospectively. The final diagnosis was established at the end of follow-up or when the patients underwent surgery or died.
Fine needle aspiration was indicated in 246 of 293 cases (84%), considered technically feasible in 232 of 246 cases (94%) and gave adequate samples for histopathological diagnosis in 204 of 232 cases (88%). Endoscopic ultrasound sensitivity, specificity and accuracy were 79, 60 and 72%, respectively; the corresponding figures for endoscopic ultrasound-fine needle aspiration were 80, 86 and 82%. There was good agreement with final diagnosis for endoscopic ultrasound-fine needle aspiration (kappa 0.673, 95%CI 0.592-0.753), greater than that for endoscopic ultrasound alone (kappa 0.515, 95%CI 0.425-0.605). There was one case of intracystic haemorrhage and one case of transient hyperthermia (0.3%).
Endoscopic ultrasound-fine needle aspiration of pancreatic masses seems to be feasible, effective and safe in this consecutive study of patients.
Digestive and Liver Disease 09/2007; 39(8):768-74. · 3.05 Impact Factor
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ABSTRACT: Evaluation of the importance of the on-site presence of a skilled cytopathologist during endoscopic ultrasound-guided fine needle aspiration at determining samples' adequacy and performing ancillary techniques which can be helpful for the diagnosis.
A retrospective analysis of our institute's experience with EUS-FNA sampling is presented. From January 2001 to May 2007, 404 patients underwent the EUS-FNA evaluation. From 2003 a cytopathologist was present during the procedure and started making an extemporary evaluation of the samples' adequacy.
Before 2003, a final cytological diagnosis was available in only 70% of the cases (without an on-site cytopathologist). After 2003, in 90% of the cases (with an on-site cytopathologist). It is possible planning and performing: immunocytochemistry on cell block material including evaluation of the proliferation index; to obtain a sample for the flow cytometry in cases of lymphomas or a microbiologic workup in cases of infective lesions.
The quality of the specimens and the proper handling of the aspirated sample are very important to succesfully obtain a definitive cytological diagnosis in EUS-FNA. On-site evaluation and triage of the material allow to improve the accuracy of the diagnosis.
Minerva medica 09/2007; 98(4):395-400. · 0.90 Impact Factor
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Luigi Cataliotti,
C de Wolf,
Roland Holland,
Lorenza Marotti,
Nick Perry,
K Redmond,
Marko Rosselli Del Turco,
Henny Rijken,
Nora Kearney,
I O Ellis, [......],
Nial O'Higgins,
E. Pennery,
D. Rainsbury,
Emiel Rutgers,
M. Smola,
E Van Limbergen,
K. von Smitten,
C Wells,
R Wilson,
EUSOMA
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ABSTRACT: According to EUSOMA position paper ‘The requirements of a specialist breast unit’, each breast unit should have a core team made up of health professionals who have undergone specialist training in breast cancer. In this paper, on behalf of EUSOMA, authors have identified the standards of training in breast cancer, to harmonise and foster breast care training in Europe. The aim of this paper is to contribute to the increase in the level of care in a breast unit, as the input of qualified health professionals increases the quality of breast cancer patient care.
European Journal of Cancer 04/2007; · 5.54 Impact Factor
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Annals of the New York Academy of Sciences 12/2006; 464(1):262 - 274. · 3.15 Impact Factor
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ABSTRACT: To reveal architectural structure and growth patterns of tubular carcinomas (TC) and tubulo-lobular carcinomas (TLC) of the breast.
We studied a series of 20 pure TC and 22 TLC, evaluating the architectural features of the two entities by bi-dimensional microscopy and by 3-D modelling. We traced the spatial organization of three TCs and three TLCs on serial sections using AE1/AE3 cytokeratin as a marker of the epithelial structures and reconstructed 3-D models of each histological type. The analysis of TC on serial cytokeratin-stained sections showed that the form of the 'tubules' was related to the plane of sectioning and that often they were tear-drop shaped, with a final tail of single cells connecting them together. 3-D models corresponded to a necklace appearance and the tubules of TC appeared as blebs bridging through solid cords to other blebs. In TLC the structure was similar, but the connecting single-cell files were usually longer. Both TC and TLC showed similar E-cadherin positivity and an indolent clinical behaviour.
TC and TLC share the same architectural and growth patterns and ultimately seem to represent variants of the same tumour type.
Histopathology 05/2006; 48(5):556-62. · 3.08 Impact Factor
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ABSTRACT: The immunohistochemical determination of HER-2 to identify patients with advanced breast cancer candidates for Trastuzumab treatment proved neither accurate nor fully reliable, possibly because none of the current reagents detects the specific antigenic site target of Trastuzumab. To circumvent this problem, we conjugated the NH2 groups of Trastuzumab with biotin, and the compound obtained, designated BiotHER, was added directly to tissue sections. Biotin-labelling was revealed with horseradish peroxidase-conjugated streptavidin. Specificity and sensitivity of BiotHER immunostaining with respect to HER-2 amplification were tested on 164 breast carcinoma samples. BiotHER staining was detected on the tumour cell membrane of 12% of all specimens and in 49% specimens with gene amplification, while absent in nonamplified tumours. Predictivity of BiotHER status with respect to the clinical outcome was analysed in 54 patients with HER-2 amplified advanced breast cancer treated with Trastuzumab plus chemotherapy. BiotHER staining, detected in 50% of tumours with HER-2 amplification, was an independent predictor of clinical outcome. In fact, BiotHER positivity was independently associated with increased likelihood of tumour response and reduced risk of tumour progression and death. Biotinylated Trastuzumab can thus be used for immunohistochemical detection of HER-2 overexpression in breast cancer, and has the potential to identify patients likely to benefit from Trastuzumab treatment.
British Journal of Cancer 05/2005; 92(7):1261-7. · 5.04 Impact Factor
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ABSTRACT: The need for further axillary treatment in patients with breast cancer with low-volume sentinel node (SN) involvement (micrometastases or smaller) is controversial.
Twenty-five studies reporting on non-SN involvement associated with low-volume SN involvement were identified using Medline and a meta-analysis was performed.
The weighted mean estimate for the incidence of non-SN metastases after low-volume SN involvement is around 20 per cent, whereas this incidence is around 9 per cent if the SN involvement is detected by immunohistochemistry (IHC) alone. Subset analyses suggest that studies with axillary dissection after any type of SN involvement result in somewhat higher estimates than studies allowing omission of axillary clearance, as do studies with more detailed histological evaluation of the SN compared with those with a less intensive histological protocol. Higher-quality papers yield lower pooled estimates than lower-quality papers.
The risk of non-SN metastasis with a low-volume metastasis in the SN is around 10-15 per cent, depending on the method of detection of SN involvement. This should be taken into account when assessing the risk of omission of axillary dissection after a positive SN biopsy yielding micrometastatic or immunohistochemically positive SNs.
British Journal of Surgery 11/2004; 91(10):1245-52. · 4.61 Impact Factor
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C A Wells,
J P Sloane,
D Coleman,
C Munt,
I Amendoeira,
N Apostolikas,
J P Bellocq,
S Bianchi,
W Boecker, G Bussolati, [......],
B Borisch,
G Cserni,
T Decker,
H Kerner,
J Kulka,
P Regitnig,
A Sapino,
A M Tanous,
S Thorstenson,
E Zozaya
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ABSTRACT: To assess the variability of oestrogen receptor (ER) testing using immunocytochemistry, centrally stained and unstained slides from breast cancers were circulated to the members of the European Working Group for Breast Screening Pathology, who were asked to report on both slides. The results showed that there was almost complete concordance among readers (kappa=0.95) in ER-negative tumours on the stained slide and excellent concordance among readers (kappa=0.82) on the slides stained in each individual laboratory. Tumours showing strong positivity were reasonably well assessed (kappa=0.57 and 0.4, respectively), but there was less concordance in tumours with moderate and low levels of ER, especially when these were heterogeneous in their staining. Because of the variation, the Working Group recommends that laboratories performing these stains should take part in a external quality assurance scheme for immunocytochemistry, should include a tumour with low ER levels as a weak positive control and should audit the percentage positive tumours in their laboratory against the accepted norms annually. The Quick score method of receptor assessment may also have too many categories for good concordance, and grouping of these into fewer categories may remove some of the variation among laboratories.
Archiv für Pathologische Anatomie und Physiologie und für Klinische Medicin 09/2004; 445(2):119-28. · 2.49 Impact Factor
