A Calibrated Multiexit Neural Network for Detecting Urothelial Cancer Cells.

Deep convolutional networks have become a powerful tool for medical imaging diagnostic. In pathology, most efforts have been focused in the subfield of histology, while cytopathology (which studies diagnostic tools at the cellular level) remains underexplored. In this paper, we propose a novel deep learning model for cancer detection from urinary cytopathology screening images. We leverage recent ideas from the field of multioutput neural networks to provide a model that can efficiently train even on small-scale datasets, such as those typically found in real-world scenarios. Additionally, we argue that calibration (i.e., providing confidence levels that are aligned with the ground truth probability of an event) has been a major shortcoming of prior works, and we experiment a number of techniques to provide a well-calibrated model. We evaluate the proposed algorithm on a novel dataset, and we show that the combination of focal loss, multiple outputs, and temperature scaling provides a model that is significantly more accurate and calibrated than a baseline deep convolutional network.

Presence of cancer cells in gastric lavage of gastric cancer patients as an indicator of advanced disease, predictor of tumour aggressive phenotype and independent prognostic factor for poor survival: The endoluminal metastatic pathway of gastric cancer and GL0/GL1 classification.

OBJECTIVE: As of 2017, the pathobiology of gastric cancer (GC) is far from fully understood; consequently, new methods of basic and advanced research have been proposed and tested. The presence (GL1) vs absence (GL0) of malignant cells exfoliated in gastric lavage (GL) of GC patients was formerly evaluated with diagnostic intent but not for staging or prognostic assessment. We investigated this hitherto unreported application of cytopathology. METHODS: GL was preoperatively and prospectively collected from 80 GC patients and cytologically analysed. The results were compared with the classic clinicopathological features of GC and related to survival. The prognostic value of GL1 was assessed through univariate and multivariate analyses. RESULTS: GL1 was detected in 36 samples (45%) and correlated with advanced tumour depth (T3-T4), lymphatic metastasis (N+), distant metastasis (M1) and lymphovascular invasion (LVI1; P=.0317, .0024, .003 and .0028, respectively). Overall survival (OS) was significantly shorter for GL1 (23 months) vs GL0 patients (42 months; P=.005) and GL1 vs GL0 T1 subjects (12.6 vs 47.8 months, P=.0029). Univariate analysis revealed that GL1, N+, M1, LVI1 and advanced stage were significantly associated with OS. Multivariate analysis assessed GL1 as the only independent prognostic factor for worse OS and progression-free survival (P=.0013 and .0107). CONCLUSIONS: In the present study, GL1 was correlated with advanced disease, aggressive tumour behaviour and poor prognosis. Although additional studies are needed to confirm these findings, the GL0/GL1 classification can be applied to GC patients to achieve higher accuracy in staging, prognostic stratification and treatment selection.

Efficacy and cost effectiveness of rapid on site examination (ROSE) in management of patients with mediastinal lymphadenopathies.

BACKGROUND: The diagnostic and staging approach for the mediastinal lymphadenopathies, with or whithout pulmonary lesions endoscopically visible, is based on transbronchial needle aspiration (TBNA) during fiberoptic bronchoscopy and on mediastinoscopy. One important factor impacting on TBNA sensitivity is the rapid on site cytological examination (ROSE). AIM: The aim of this study was to evaluate the economic impact of TBNA and TBNA + ROSE, in the diagnosis of these lesions. PATIENTS AND METHODS: 120 patients, affected by mediastinal lymphadenopathies suspected for lung cancer, underwent TBNA during fiberoptic bronchoscopy: 60 patients without ROSE (group A) and other 60 with ROSE (group B). Whenever needle aspirations failed to provide diagnosis, the patient underwent mediastinoscopy. The economic impact of the diagnostic process was performed. RESULTS: In group A, 39 patients (65%) obtained a diagnosis with TBNA while 21 patients (35%) required mediastinoscopy. In group B, 48 patients (80%) obtained a diagnosis with TBNA + ROSE, while 12 patients (20%) required mediastinoscopy. With regards to the costs of the procedures performed in the diagnostic process, the use of TBNA with ROSE as first diagnostic approach has saved a considerable amount of euros (19,413) compared to the use of TBNA without ROSE and the combined procedure increased (p < 0.02; chi square test) the sensitivity of TBNA by 15%. CONCLUSIONS: ROSE significantly impacts on the diagnostic yield, as well as on the overall management costs of patients with mediastinal lymphadenopathy, suspected for lung cancer.

Peritoneal wash cytology in gastric carcinoma. Prognostic significance and therapeutic consequences.

BACKGROUND AND AIMS: The prognosis of patients with gastric cancer is poor, even following curative resection, and is related primarily to the extent of disease at presentation. In locally advanced gastric tumors, peritoneal lavage cytology (PLC) is a relevant prognostic factor. The Authors present their results of peritoneal washing cytology, evaluating the prognostic value of this technique, and discussing the clinical impact. PATIENTS AND METHODS: From July 2003 to May 2008, results of PLC in 64 patients with histologically proven primary gastric adenocarcinomas were analyzed. At laparotomy the abdomen was irrigated with 200 ml of normal saline, and ≥50 ml were aspirated and examined by means of cytology and immunocytopathology. RESULTS: PLC was positive in 7 cases (11%). Overall, 86% of patients with a positive PLC had a pT3/pT4 tumor and 100% with a positive PLC had an N-positive tumor (p < 0.001); 71% of patients with a positive PLC had a grade G3/G4 tumor (p = 0.001). At a median follow-up of 32 months, the cumulative 5-year survival was 28%. The median survival of patients presenting positive PLC (19 months) was significantly lower than that of patients with negative peritoneal cytology (38 months) (p = 0.0001). Multivariate analysis identified cytology as a significant predictor of outcome (p = 0.018). CONCLUSIONS: Results in the present series demonstrated that patients with a positive peritoneal cytology had advanced disease and poor prognosis, thus indicating that patients with locally advanced gastric cancer should undergo staging laparoscopy and PLC examination in order to select those requiring more aggressive treatment. Future therapeutic strategies should include PLC examination in preoperative staging, in order to select patients for more aggressive treatment.

[Detection, characterization and clinical significance of circulating cancer cells in patients surgically treated for breast cancer]. / Ricerca, caratterizzazione e significato clinico di cellule tumorali circolanti in pazienti operate per cancro della mammella.

Recent technological advances have led to an increasing ability to detect isolated or groups of tumour cells in blood, lymph nodes or bone marrow in patients with different tumour types. However, the clinical evidence of these advances is unclear. The detection and the characterisation of circulating breast cancer cells and the eventually micrometastasis represent an important prognostic factor with therapeutic implications. The number of neoplastic cells being very small, these are not easily detected by using only cytomorphology, possibly associated to immunocytochemistry. In the last decade many studies have been directed in order to identify new assays. In the present review the Authors summarize advantages and disadvantages about two different technical approaches: molecular and immunomagnetic selection with cellular enrichment and immunocytochemistry.

[Changes in the expression of cellular alpha and beta tubulins in patients with sporadic type colorectal cancer]. / Alterazione della espressione delle tubuline cellulari alfa e beta in pazienti affetti da cancro colorettale di tipo sporadico.

The aim of this preliminar report is to evaluate alfa and beta tubulins, components of cellular microtubules, alterated expression in sporadic colorectal cancer patients. The Authors considered 16 patients who underwent surgery for sporadic colorectal carcinoma with radical intent. Alfa and beta tubulins were evaluated in tumoral mucosa by immunohistochemistry. In 56.2% of the examined patients a low expression of alfa and beta tubulins was showed while the alteration of alfa tubulin was showed in 81.2% of the patients. This finding supports the hypothesis of Porter that alterations in microtubule structure might be part of the cellular response to DNA damage.

[From molecular biology to new treatment approaches to colorectal cancer: basic research, experimental trials and surgical implications]. / Dalla biologia molecolare ai nuovi approcci terapeutici del cancro colorettale: ricerca di base, sperimentazione clinica ed implicazioni chirurgiche.

The Authors review the natural history of colorectal cancer from the point of view of molecular biology and genetics from aberrant crypts foci and familiar adenomatous polyposis to hereditary non polyposis colon cancer and sporadic colorectal cancer. They carry out international literature about basis knowledges, experimental trials and personal studies. Up to day traditional colorectal cancer surgical treatments and adjuvant or neoadjuvant pharmacological therapy cannot be modified, nevertheless "new drugs generation" known as signal transduction inhibitor could, in the future, prove to be an effective cancer treatment. The Authors highlight recent experimental clinical trials probably able to prevent sporadic colorectal cancer spreading and precursor evolution.

CD44v6 and Nm23-H1 protein expression related to clinico pathological parameters in colorectal cancer.

OBJECTIVE: Nm23-H1 and CD44v6 expression has been shown to be correlated with the metastatic potential of colorectal cancer (CRC) in some studies but not in others. The present study was undertaken to evaluate immunohistochemically the expression of these markers and to correlate them with clinicopathological variables. MATERIALS AND METHODS: Archival tissues of 41 non metastatic colorectal cancers were histopathologically evaluated and stained with monoclonal antibodies versus Nm23-H1 and CD44v6. RESULTS: Expression of Nm23-H1 was detected in 73% (n = 30) of all CRC, and CD44v6 in 37% (n = 15) of all CRC. CD44v6 was found to be statistically associated with tumour grading differentiation (p < 0.03), but no correlation emerged between Nm23-H1 and CD44v6 and Dukes stage, site, peritumoral lymphocytic infiltration, venous infiltrating, perineural infiltrating, tumour budding, pushing and infiltrating tumour growth. CONCLUSION: Even if the results are not statistically significant, the authors noticed that the expression of Nm23-H1 was correlated with those histopathological parameters that indicate local disease progression and metastases.

[Is the Lauren classification an independent parameter for the prognostic evaluation of surgically treated gastric cancer patients? Analysis of a case series]. / La classificazione di Lauren può essere considerata un parametro indipendente per la valutazione prognostica di pazienti operati per cancro gastrico? Analisi di una casistica.

The Authors highlight the efficacy of the Lauren's classification in 28 surgically treated gastric cancer patients. Lauren's classification allows a prognostic evaluation corresponding to the effective gastric cancer natural history. Present histo-morphological classification criteria appear not to coincide with the clinical evolution; as a matter of fact over- or understaging is possible in gastric cancer patients. 64,28% of the Lauren's classification intestinal type patients survive after a four year follow up vs. 42,85% of the diffuse type patients. The Authors discuss about new biomolecular knowledge in gastric cancer oncogenesis.

Brain metastasis from hepatocellular carcinoma associated with hepatitis B virus.

Four cases of brain metastasis from hepatocellular carcinoma (HCC) associated with hepatitis B virus (HBV) are reported in an area not endemic for HBV infection. Two cases are unusual, since cerebral metastases were the only secondary localization. In these cases, no other sites of metastasization were detected either before or immediately following neurosurgical treatment. In all cases the expression of pRB, p53 and p16 tumor suppressor protein was studied with immunohistochemistry, both, in the primary and metastatic lesions. The pRB expression was as follows: in two cases, lack and moderate expression were observed both, in the primary and in the metastases; in the other two, pRB was not detected. In all cases p53 expression was negative both, in the primary and the metastases. P16 expression was moderately expressed in three cases, both in the primary and the metastases. In one case it was absent. Hepatocarcinogenesis is a multistep process, in which several oncogenes and oncosuppressor genes are involved. In four unusual cases of spread to the brain, we evidenced that tumor suppressor protein expression of p16, p53, and particularly pRB (its aberrated expression is usually associated with metastasis) were altered. We also suggest that HBV and its X protein (HBX) might play an important role in such aggressive behavior of the neoplasia.

[Genetics-based prognosis evaluation of patients surgically treated for sporadic colorectal cancer]. / Valutazione prognostica su base genetica di pazienti operati per cancro colorettale di tipo sporadico.

The basic assumption as rationale of this research was that DNA repair genes (MMR system) are at beginning of the genetic mutational cascade causing the induction of oncogenesis of sporadic colorectal cancers as well as their multiclonal heterogeneity. In a previous study the Authors randomly selected, from a series of 256 patients, 29 patients up to the age of 60 years who underwent surgery for colorectal carcinoma with radical intent. All selected cases were considered as sporadic cancers from a clinical point of view, since none of them fulfilled the Amsterdam criteria for HNPCC and familial adenomatous polyposis was included too. Mismatch repair gene proteins expression and, in particular, gene hMSH2 protein was investigated by immunohistochemistry analysis. In 12 cases (41.4%) hMSH2 exhibited strong expression in the tumoral cells as well as in the surrounding mucosa and at distant mucosa. In 14 cases (48.3%) loss of hMSH2 protein expression was observed in tumoral cells and low immunoreactivity was detected in peritumoral mucosa while strong hMSH2 expression was observed in distant mucosa. In a third small group of patients (10.3%) loss of hMSH2 protein expression was detected in tumoral, adjacent and at distance normal mucosa. After a five years follow up, 100% of twelve patients of first group are still alive vs 64.3% of fourteen patients of second group, while in the third group only one patient survives. These results support the hypothesis of an involvement of hMSH2 gene defect in development of a subset of sporadic colorectal cancer. For the patients with strong expression of hMSH2 in the tumoral cells as well as in the surrounding mucosa and at distant mucosa, this parameter could represent an independent criterion for a good prognostic value.

Msh2, Mlh1, Fhit, p53, Bcl-2, and Bax expression in invasive and in situ squamous cell carcinoma of the uterine cervix.

To analyze relevant factors and their effects on neoplastic progression in cervical carcinoma, a panel of genetic markers was studied. Paraffin-embedded tissue sections were obtained from 37 patients with carcinoma of the uterine cervix, 14 noninvasive squamous cell carcinomas (NISCCs), and 23 invasive squamous cell carcinomas (ISCCs). Immunoreactivity of Msh2, Mlh1, Fhit, p53, Bcl-2, and Bax proteins was examined by immunohistochemical staining with appropriate antibodies. Positive staining of Msh2 was detected in 13 of 14 (92.9%) NISCCs and in 13 of 23 (56.5%) ISCCs (P < 0.02). Mlh1 immunoreactivity was observed in 10 of 14 (71.4%) NISCCs and in 8 of 23 (34.8%) ISCCs (P < 0.04). Overexpression of p53 protein was found in 4 of 14 (28.6%) NISCCs and in 16 of 23 (69.6%) ISCCs (P < 0.02). Bcl-2 overexpression was detected in 2 of 14 (14.3%) NISCCs and in 15 of 23 (65.2%) ISCCs (P < 0.003). No significant difference in the two types of lesion was found for Bax and Fhit expression. The relationship between Mlh1, Msh2, and p53 protein expression was significant (P < 0.001 and P < 0.001, respectively), as was that between Fhit and Bax immunoreactivity (P < 0.02). In conclusion, we consider that altered expression of Msh2, Mlh1, p53, and Bcl-2 may be a critical event during cervical cancer progression, whereas Fhit may be a component of a proapoptotic pathway.

nm23-H1 protein expression in anal canal carcinoma: does it correlate with prognosis?

BACKGROUND AND OBJECTIVES: Anatomic extent is not the sole axis of classification of tumors and of tumor patients relevant to treatment planning and estimation of prognosis. This results in the need to demonstrate an improvement in prognostic assessment and choice of therapy achieved by consideration of factors other than TNM. nm23 protein does prevent tumor from metastasizing and may also play a role in the control of growth and development. The purpose of this study was to elucidate the clinical significance of nm23 expression in human anal canal carcinoma and to evaluate its influence on the outcome of patients after surgery or radiochemotherapy. METHODS: Twenty-two patients affected by anal canal carcinoma were evaluated. Each section was incubated with monoclonal antibody nm23 NDPK-A. Immunostaining was considered positive when at least 10% of the tumor cells were immunostained. RESULTS: nm23 immunoreactivity was detected in 6/22 (27.3%) tumors. No significant association was found between nm23 expression and prognosis. CONCLUSIONS: The mechanisms causing enhanced nm23-H1 expression in anal canal carcinoma are unknown. Although the level and expression were not correlated with prognosis, activation of nm23-H1 gene might be a prerequisite for oncogenesis in this type of tumor, while an alternate possibility is the modification of cellular characteristics in relation to proliferation and/or differentiation as a consequence of oncogenesis.

BCL2 and BAX expression in hyperplastic and dysplastic rectal polyps.

BACKGROUND/AIMS: Members of the gene family that includes BCL2 and BAX are functionally antagonists in the apoptosis process and they have been observed in normal and neoplastic tissues. The aim of this study is to investigate the combined effects of BCL2 and BAX protein in normal mucosa, dysplastic and hyperplastic polyps of the rectum. METHODOLOGY: We studied BCL2 and BAX protein expression in 40 cases of adenomatous polyps all located in the rectum, with different dysplastic gradings, and the mean time in 10 cases of normal rectal mucosa. RESULTS: BCL2 expression was found more frequently in hyperplastic and in low dysplastic polyps with moderate and strong positivity compared to moderate and severe dysplasia. BAX expression was found in normal mucosa in hyperplastic and dysplastic polyps, the immunoreactivity was prevalently moderate and strong. CONCLUSIONS: These preliminary data suggest that BCL2 and BAX confirm a probably different role in apoptosis. Nevertheless, it is important to know the relation between the molecular pathways of apoptosis, the defective mismatch repair and the tumor suppressor genes associated with an increased mutation rate in cancerogenesis of the colorectum.

Altered expression of hMSH2 in sporadic colorectal cancer, surrounding mucosa and at distant colonic mucosa.

BACKGROUND: Mismatch repair gene hMSH2 is involved in correction of mispairing during replication and its mutation is associated both with microsatellite instability and with hereditary colorectal cancer. We evaluated its involvement in sporadic colorectal cancer tumorigenesis too. MATERIALS AND METHODS: The protein expression pattern of hMSH2 was evaluated on 29 cases of resected sporadic adenocarcinoma using an immunohistochemical approach. RESULTS: In 14 cases, lack of hMSH2 protein expression was observed in adenocarcinoma and in peritumoral mucosa. In 12 patients, hMSH2 resulted in strong expression in the tumour as well as in the surrounding mucosa and at distant mucosa. In three cases, hMSH2 protein expression in tumoral, adjacent and at distance normal mucosa resulted negative. CONCLUSION: Repair genes could play an important role in tumour progression and in sporadic colorectal cancer. Detection of protein expression by immunohistochemistry may be a method to select tumours for successive genetic investigations.

Correlation between nm23-H1 overexpression and clinicopathological variables in human anal canal carcinoma.

To improve life expectancy prognostic factors other than TNM have been investigated. It is thought that nm23 protein may play a specific biological role in suppressing tumor metastasis. The purpose of this study was to elucidate the clinical significance of nm23 expression in human anal canal carcinoma. Immunostaining using anti-nm23 monoclonal antibody was performed in 22 anal canal tumors. The results were correlated with clinicopathological variables. Six cases out of 22 (27.3%) were nm23-positive. Significant association was found between nm23-H1 expression and depth of invasion, lymph node involvement and prognosis (p<0.05). There was no significant association between nm23-H1 expression, histologic type and age of the patients. nm23-H1 expression was not seen in our cases with metastasis and this may be related to nm23 gene alterations not being detectable by the monoclonal antibody used or to the presence of a subset of tumors in which nm23 gene abnormalities had not yet occurred at the time of tumor excision or biopsy. Overexpression of nm23-H1 protein in anal canal carcinoma may have implications for its metastatic potential. nm23-H1 expression would provide a more accurate evaluation of outcome for individual patients and thus improve treatment planning.

[Genetic factors and colorectal cancer: a new approach to clinical experimentation]. / Fattori genetici e cancro del colon-retto: una nuova frontiera per la sperimentazione clinica.

The Authors point out the basis for a better characterisation of colo-rectal cancer and precursory lesions. In fact the etiology of familial adenomatous polyposis (FAP), aberrant crypt foci (ACF), hereditary non polyposis colon cancer syndrome (HNPCC) seems to be correlated to molecular pathology. Therefore the Authors review colo-rectal cancer natural history which frequently appears to be not related to clinical evolution.

Human papillomavirus infection and p53 nuclear overexpression in anal canal carcinoma.

The product of HPV E6 and E7 genes is able to inactivate both the p53 and pRb proteins. The aim of this study was to evaluate the correlation among anal HPV infection and nuclear p53 overexpression. The Authors evaluated HPV DNA by PCR and p53 nuclear expression by immunohistochemistry in 12 cloacogenic and 6 squamocellular carcinoma. HPV DNA was detected in 71.4% of the squamocellular tumors and in 57.1% of the cloacogenic tumors. In squamocellular tumors HPV types 31-33 and 16 were found; in cloacogenic tumors type 16 alone was detected. Nuclear accumulation of p53 was found to be associated with the presence of HPV. There was no significant difference in parietal infiltration, lymph nodes involvement and prognosis between HPV+p53+ patients and HPV-p53- patients. Tumor aggressiveness is likely to be enhanced by factors other than HPV infection and p53 overexpression.

Intratumoral heterogeneity in colorectal carcinoma: trucut sampling for DNA ploidy analysis.

BACKGROUND: Solid tumors, such as colorectal carcinomas, consist of cell subpopulations that differ both genetically and in their clinical behavior. Many authors have examined cell kinetics and DNA content in colorectal tumors in correlation to clinical and pathological variables with different results. The interpretation of those results present some difficulties related to tumor heterogeneity that to date are unsolved. Our study is based on a new method of colon cancer sampling for DNA content determination. The aim of this work was to reduce the risk of incorrect DNA evaluation due to tumor heterogeneity. MATERIAL AND METHODS: Our study was based on eleven selected cases of T3 colorectal carcinoma. Fresh surgical specimens from the primary tumor site were taken during surgery. For each case at least four samples were taken using a 23 gauge trucut from the outside of the serosa through the tumor to the lumen of the colon. The specimens were stained according to a modified Feulgen method and DNA content was measured by image analysis. Three parameters were evaluated: DNA index, ploidy and proliferation level (considered as the sum of elements corresponding to the S and G2 phases). RESULTS: One of the eleven (9.1%) tumors showed a diploid pattern; four out of eleven (36.4%) cases showed a tetra/polyploid pattern and six out of eleven (54.5%) cases showed an aneuploid pattern. Three tumors were monoclonal (27.3%), one diploid and two aneuploid. Eight were polyclonal (72.7%). Considering the single specimen, seven out of sixty-eight specimens (10.3%) were inadequate because of scanty material. Twenty-five out of the sixty-one adequate specimens (41%) were monoclonal and thirty-six (59%) were polyclonal. Five tumors (three monoclonal and two polyclonal) showed the same cell clones on all the examined samples. The remaining six tumors showed interregional variability. The six of the eight polyclonal cases (75%) multiple stem lines were evident, analyzing only one sample taken close to colon serosa, while in one case (25%) it was necessary to examine two samples in order to see the polyclonality of the lesion. When samples taken close to mucosa where analyzed, however, one sample was not enough to show tumor polyclonality in five of the eight polyclonal examined cases. Proliferation level varied greatly in different parts of the same carcinoma and did not correlate to the site from which the sample was taken. CONCLUSION: In the present study, we demonstrated that DNA ploidy differences may exist between the superficial and the deep part of the same neoplasia and that tumor samples show a greater variability in the deeper layers. Using trucut samplings, it was possible to point out the majority of aneuploid cell populations close to the serosa. In conclusion, trucut biopsy permits full thickness sampling of the tumoral mass and allows, from few samples, to evaluate the multiple DNA stemlines present in different parts of a colorectal tumor.