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.

Stearoyl-CoA desaturase-1 is a key factor for lung cancer-initiating cells.

In recent years, studies of cancer development and recurrence have been influenced by the cancer stem cells (CSCs)/cancer-initiating cells (CICs) hypothesis. According to this, cancer is sustained by highly positioned, chemoresistant cells with extensive capacity of self renewal, which are responsible for disease relapse after chemotherapy. Growth of cancer cells as three-dimensional non-adherent spheroids is regarded as a useful methodology to enrich for cells endowed with CSC-like features. We have recently reported that cell cultures derived from malignant pleural effusions (MPEs) of patients affected by adenocarcinoma of the lung are able to efficiently form spheroids in non-adherent conditions supplemented with growth factors. By expression profiling, we were able to identify a set of genes whose expression is significantly upregulated in lung tumor spheroids versus adherent cultures. One of the most strongly upregulated gene was stearoyl-CoA desaturase (SCD1), the main enzyme responsible for the conversion of saturated into monounsaturated fatty acids. In the present study, we show both by RNA interference and through the use of a small molecule inhibitor that SCD1 is required for lung cancer spheroids propagation both in stable cell lines and in MPE-derived primary tumor cultures. Morphological examination and image analysis of the tumor spheroids formed in the presence of SCD1 inhibitors showed a different pattern of growth characterized by irregular cell aggregates. Electron microscopy revealed that the treated spheroids displayed several features of cellular damage and immunofluorescence analysis on optical serial sections showed apoptotic cells positive for the M30 marker, most of them positive also for the stemness marker ALDH1A1, thus suggesting that the SCD1 inhibitor is selectively killing cells with stem-like properties. Furthermore, SCD1-inhibited lung cancer cells were strongly impaired in their in vivo tumorigenicity and ALDH1A1 expression. These results suggest that SCD1 is a critical target in lung cancer tumor-initiating cells.

Colostrum in menopause effects on vaginal cytology/symptoms.

The aim of this study was to assess the effects of three weeks of daily colostrum cream on vaginal cytology and local symptoms related to menopause. Genito-urinary symptoms and cell morphology were analyzed at time 0 (T0) and after three weeks (16 +/- days since the end of treatment) at time 1 (T1). Dyspareunia, vaginal dryness, and maturation index (MI) reached a statistically significant difference between T0 and T1. The results proved to be an alternative treatment for vaginal distress caused by lack of hormones in patients in which hormonal treatment is contraindicated.

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.

Prognostic implication of high risk human papillomavirus E6 and E7 mRNA in patients with intraepithelial lesions of the cervix in relationship to age.

Since the introduction of the cytological screening programs, a significant reduction in the incidence of cervical cancer has been achieved. Almost all of these cancers are related to high-risk (HR) Human Papillomavirus (HPV) cervical infections. However, the natural history of HPV infection seems to be different in younger patients, resulting in a higher rate of regression. There is, therefore, the need to identify HPV-related biomarkers in order to enhance the effectiveness of screening of high-risk cytological lesions, in particular in women over 35 years of age. This study aims to evaluate the prognostic value of the HR HPV E6 and E7 mRNA expression in women with intraepithelial lesions of the cervix, older or younger than 35 years of age. One hundred and eighty-four HR HPV DNA positive patients with a low squamous intraepithelial lesion (LSIL) were tested for mRNA expressions, included in an observational study, and evaluated at follow-up with standard cytology up to 24 months from the mRNA test. The frequency of HSIL/LSIL cytology in the older cohort of mRNA positive patients was significantly higher compared to mRNA-negative patients, both at 1 and 2 years of follow-up (Chi-square: p 0.007 and p 0.009), but this difference was not found in the younger cohort. According to our results, the E6/E7 mRNA test could be a biomarker for viral activity, useful in identifying patients at higher risk of abnormal cytology, and in implementing the management of HR HPV DNA-positive women over 35 years of age.

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.

Burden of HIV/AIDS infection before and during the civil war in Somalia.

Somalia has suffered a massive internal population displacement and exodus that began in 1988 and is still ongoing during the prolonged and intermittent civil war. This review looks at the burden of HIV infection in Somali and the impact of civil war on its epidemiology. Serosurveys have indicated that HIV was not present in Somalia before the civil war and to date Somalia has had an HIV prevalence markedly below that of its neighbours. However, due to the ongoing war HIV sentinel surveillance cannot reach most of the affected areas in Somalia and the current HIV infection problem may be greater than the figures indicate.

Burden of HIV/AIDS infection before and during the civil war in Somalia

Somalia has suffered a massive internal population displacement and exodus that began in 1988 and is still ongoing during the prolonged and intermittent civil war. This review looks at the burden of HIV infection in Somali and the impact of civil war on its epidemiology. Serosurveys have indicated that HIV was not present in Somalia before the civil war and to date Somalia has had an HIV prevalence markedly below that of its neighbours. However, due to the ongoing war HIV sentinel surveillance cannot reach most of the affected areas in Somalia and the current HIV infection problem may be greater than the figures indicate

E-learning for laurea in biomedical laboratory technicians: presentation of a pilote study.

The aim of the paper is to afford the design and construction of an e-learning model answering to these requirements for the LHCP in Technician of Biomedical Laboratory. The SCENARIO of work for the technician in biomedical laboratory (TBL) is radically changed and e-learning should answer to these new challenges. In particular today He or She should be able to mange in tele-pathology applications the tissue on the so called virtual glass. An architecture for the e-learning based on Web-Dav and a Light software for the virtual glass tele-pathology consultation has been designed with also a special care for the teachers' and students' platforms. At the moment we are investigating Telepathology platforms for the extranet consulting of virtual glasses.

[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.

[New and old procedures in the diagnosis of dubious breast lesions]. / Procedure diagnostiche tradizionali e di recente introduzione nelle lesioni mammarie di dubbia interpretazione.

In the last years the detection of early breast cancers (lesions less than one centimetre in diameter, with good prognosis) has consistently increased for the wide application of mammary screening programs. At the same time, an increasing number of radiographically detected unexpected lesions (nonpalpable breast lesions) has been evidenced. In those cases, often both mammography and ultrasound evaluation are dubious and a multidisciplinary diagnostic approach is mandatory. Fine-needle aspiration (FNA) and core biopsy (CB) are well established diagnostic methods but, in recent years, new microinvasive bioptic procedures (as the Mammotome and the ABBI systems) have been introduced. In this review the limits and the possibilities of the classical and new cytohistological techniques are evaluated. A possible multistep diagnostic approach is described on a cost-benefit basis and in consideration of the various procedures.

Immunocytochemical expression of Ki67 and laminin in Hurthle cell adenomas and carcinomas.

BACKGROUND: Hurthle cell neoplasms may occur as a benign adenoma or carcinoma; the latter displays comparatively aggressive clinical behaviour. Fine-needle aspiration cytology (FNAC) represents a reliable tool for screening Hurthle cell lesions before surgery. Nevertheless, the cytological interpretation of these lesions is not always unequivocal. We analyzed cell growth and cellular adhesiveness by means of two different antibodies, Mib1 (Ki67) and laminin. AIM: The aim of the study was to analyze Ki67 and laminin immunocytochemical expression on FNAC to evaluate their usefulness in the preoperative differential diagnosis of Hurthle cell neoplasms. RESULTS: A higher expression of Ki67 has been recorded in malignant lesions as compared to benign ones (8% to 20% vs 1% to 5% nuclear staining, respectively; p < 0.001). An increased reactivity of anti-laminin antibody was recorded in the cytoplasm of cells from all malignant lesions. In Hurthle cell adenomas this adhesion molecule showed an intensity ranging from low to moderate. Moreover, a few benign lesions showing a moderate proliferative activity were associated with evident laminin expression. CONCLUSION: These findings support the hypothesis that benign Hurthle cell lesions with a high cellular proliferation associated with an increased laminin expression could define a subset of lesions prone to malignant transformation. Conversely, since all cases with low expression of both laminin and Ki67 always correspond to adenomas, we suggest that the different expression of these two antibodies on FNAC can provide a further tool for the preoperative identification of lesions at low risk of malignancy, thus avoiding unnecessary surgery.

Study of viral integration of HPV-16 in young patients with LSIL.

AIMS: To investigate the physical status of human papillomavirus 16 (HPV-16) in low grade squamous intraepithelial lesions (LSILs) as a means of determining the percentage of viral integration. METHODS: Ninety two LSIL/HPV positive Thin Prep(TM) samples were initially tested for the E6 gene by the polymerase chain reaction (PCR) to identify the HPV-16 virus. To avoid false positive results, the specificity of the bands obtained from PCR was confirmed by Southern blot hybridisation with internal oligonucleotide probes. Next, a PCR screen for the E2 gene was performed to identify those samples in which the virus was integrated. Viral integration was detected in just over half of them. RESULTS: Twenty of the 92 samples were HPV-16 positive, as shown by PCR for the E6 gene. Southern blot analysis confirmed that 13 of these samples were positive for the viral E6 gene. Thus, viral integration was detected in just over a half of the samples positive for HPV-16. CONCLUSIONS: These data show that HPV-16 integration occurs in a subset of LSILs. The measurement of HPV-16 integration would be a helpful complementary tool for cytological evaluation in primary cervical screening to identify those patients at risk of developing high grade squamous intraepithelial lesions and cervical cancer.

Papnet-assisted cytological diagnosis intensifies the already marked variability among cytological laboratories.

OBJECTIVE: The main objective was to assess the sensitivity, specificity and reliability of PAPNET-assisted diagnosis in comparison with conventional screening. SETTING: Seven Italian and one English University or Research Institutes, and a random sample of an other 20 Italian Laboratories of the Italian National Health Service (INHS) provided the cervical smears. METHODS: During the training phase every center examined in rotation four sets of slides for a total of 300 representative slides. Afterwards, 900 "positive" slides were added to the 3,100 slides which were collected consecutively without any selection or exclusion. The eight main centers were divided into four couples and each couple of centers examined 775 slides with the PAPNET system, "blindly" to the original diagnosis. An expert cytopathologist (M.A.) of the National Institute of Health (NIH) reassessed 40% of the slides with an original negative diagnosis to evaluate the false negative rate. Two expert NIH cytopathologists (M.A., G.M.) re-examined all slides where a disagreement had been observed between the original and one or both of the study diagnoses. The main analyses concerned the following three main categories: WNL and unsatisfactory for evaluation; ASCUS, AGUS and LSIL; HSIL and carcinoma. A special algorithm was devised to define the reference diagnosis for sensitivity and specificity assessment. RESULTS: Laboratories, even belonging to the same couple, classified as "no review" a very different proportion of slides ranging from 35% to 74%. The index of kappa agreement between the members of couples examining the same sets of slides was low or very low, ranging from 0.30 to 0.03. The sensitivity of the review classification was particularly low in some laboratories. Surprisingly, only a small correlation was observed between the sensitivity of the review classification and the proportion of slides classified as "review". The "tentative" diagnosis on PAPNET tiles of the "review" slides was almost as reliable as the microscopic diagnosis. In the overall performance, there were many significant differences among the eight laboratories. The best laboratory had a sensitivity of 95% and a specificity of 96%. At least three laboratories displayed unacceptably low sensitivity and one a very low specificity. CONCLUSION: Altogether these results seem to confirm that there are wide differences among cytological laboratories per se, and that these differences are intensified by the use of an instrument like PAPNET. The huge variation in performance may be explained by differences in basic skills and by different training, but it is difficult to understand exactly what could have been done to reduce it.

Abnormal "low grade" transformation zone: current diagnostic gold standard.

The aim of this work was to examine different methods of investigation in the diagnosis of the abnormal "low grade" transformation zone of the portio. Over a period of one year 41 patients subjected to colposcopic examination underwent exo-endocervical sampling for oncologic evaluation and for detection of viral and bacterial infections (HPV, HSV, adenovirus, mycoplasmas and chlamydia trachomatis), as well as portio biopsy. A 65.8% correlation was found between cytology and the HPV-DNA test results, while histology and the presence of the HPV virus agreed in 51.4% of cases. In those cases in which minimal histological alterations were found (koilocytosis) a high percentage of HPV negativity was found. In discordant negative cytologic tests that were however positive for HPV by PCR, the genotypes identified were always 6 and 11.

Thyroiditis and oncocytic carcinoma: incidental association? A case report.

BACKGROUND: Several studies have reported an association between Hashimoto's disease and thyroid carcinoma although the cause/effect relationship is still controversial. CASE REPORT: In this paper we report the case of a 38-year-old female who first presented with a clinical history of Hashimoto's thyroiditis. Ultrasound examination showed a diffuse thyroid irregularity more pronounced in the right lobe. FNAC (Fine Needle Aspiration Cytology) was performed in this area, and the cytological diagnosis was "Hashimoto's thyroiditis". The patient underwent clinical follow-up. Two years later, an ultrasound examination showed a nodular lump in the area previously aspirated. A new FNAC evidenced a Hurthle cell neoplasia. Therefore, the patient underwent surgery. The histological diagnosis was "Hurthle cell carcinoma". CONCLUSION: In the present case, the clinical and ultrasound history suggest the development of a malignant lesion strictly related to thyroiditis, as previously reported by other studies on papillary carcinoma. A cause-effect relationship between chronic lymphocytic thyroiditis and oncocytic neoplasia is still a matter of controversy. Therefore, patients suffering from chronic thyroiditis require a careful follow-up and, in case of nodules development, FNAC is recommended.

[The significance of "atypical metaplasia" in the follow-up of patients operated for bladder cancer]. / Il significasto della "metaplasia atipica" nel follow-up dei pazienti operati per carcinoma vescicale.

BACKGROUND: The aim of this study was to evaluate the clinical usefulness of urinary cytology in the follow-up of patients who under-went surgery for bladder cancer. Particularly, the positive predictive value of urinary cytology and time elapsed between a positive test and the diagnosis of a cystoscopically confirmed bladder tumor are analyzed. METHODS: This study was carried out at the Cytological Laboratory Department of Experimental Medicine and Pathology, University La Sapienza, Rome. Among 230 cases studied since 1996, 30 male patients over 50 were examined (25 with a previous bladder cancer and 5 with a previous prostate cancer) with long time follow-up, who underwent more than two cytological examinations on voided urine (2-12) at pre-fixed intervals. RESULTS: Nine (30%) of the patients suffered from recurrent disease. The cytological examinations was positive in 8 out of the 9 positive cases and negative in 17 out of the 21 negative cases. Absence of disease was confirmed in all the latter cases both by cystoscopic examination and clinical follow-up. One negative case showed clearly malignant cells in more than one specimen taken at different time intervals. This patient is actually under strict control. In 5 cases atypical metaplasia was present in the cytological specimen. In two of those cases cystoscopic examination 5 and 8 months later confirmed progressive disease. In the other three cases cystoscopy showed no evidence of disease. Two of the patients are well and alive after 2 and 14 months respectively. One is dead of prostatic cancer. CONCLUSIONS: Considering the cases of atypical metaplasi as positive the cytological examinations showed 100% sensibility, 81% specificity, a predictive negative value of 1 and a predictive positive value of 0.66.

[The STF Project: Female Tumor Screening]. / Il progetto STF--Screening Tumori Femminili.

A pilot study has been carried out to evaluate women's compliance to a screening program for cervical cancer. The study, initiated in 1994, was performed in conjunction with the ACRO project of the CNR by the Italian National Health Institute, in collaboration with La Sapienza University of Rome and the National Institute for Cancer Research in Genoa. A preliminary telephone survey was carried out on a sample of 400 women (200 in Rome and 200 in Genoa) to assess, among other factors, their attitude towards the screening program. Afterwards, an ad hoc advertising campaign was launched and 21,827 women, randomly chosen from the register office's lists, were sent a personal invitation to participate in the screening. Most women showed interest in attending the screening program at the interview, but the percentages of participation were low (25.7% in Genoa and 27.3% in Rome). On the other hand, a high percentage of women who participated in the screening had already had a Pap test in the previous three years (Genoa, 73%; Rome, 76%). The recruiting techniques that were used in this study, and that are commonly used, do not seem to reach the core of the target population for cervical screening, i.e. women who have never had a Pap test or who had a Pap test more than 5 years earlier. New methods of recruiting aimed at categories at risk and based more on direct contacts need to be developed.

Variation in the assessment of adequacy in cervical smears.

OBJECTIVE: To assess the interobserver reproducibility of the diagnosis of 'adequacy' of cervical smears according to the Bethesda System criteria in cervical smears. STUDY DESIGN: 358 cervical smears were obtained from three Italian cytopathological centres in 1998-99. All centres provided consecutively collected smears. The cervical smears were independently and blindly assessed by four cytologists. The screening was performed using a 10x objective and an additional evaluation of the percentage of cellularity was performed using a 4x objective. RESULTS: The proportion of smears assessed by the four cytologists as 'adequate' ranged from 60% to 70%, the proportion of 'satisfactory for evaluation but limited by' ranged from 27% to 38%, and the proportion of 'inadequate smears' ranged from 2% to 4%. Full agreement in the assessment of smear adequacy was observed in 311 slides and disagreement was observed only in 47. The category 'inadequate smear' was less reliable than the other two; however, the kappa value observed was acceptable. CONCLUSION: The present study shows that it is possible to achieve a high reproducibility in the assessment of smear adequacy, at least among expert cytologists who follow the Bethesda System criteria strictly.

8-hydroxy-2'-deoxyguanosine in cervical cells: correlation with grade of dysplasia and human papillomavirus infection.

In this study, the 8-hydroxy-2'-deoxyguanosine (8-OHdG) level was assessed in human cervical cells by an immunoperoxidase method and was related to the presence of human papillomavirus (HPV) infection and precancerous lesions. After optimizing the immunohistochemical method of detecting oxidative DNA damage in whole cells, we have used this technique to estimate the oxidative damage in cervical cells collected during a routine PAP test. The analysis of variance (ANOVA) of the data from human samples showed significant differences in the 8-OHdG content among normal, low-grade and high-grade squamous intraepithelial lesion (SIL, HGSIL and LGSIL, respectively; P < 0.001). In the comparison of the three groups, statistically significant differences were detected between normal SIL and HGSIL (P < 0.001) and between LGSIL and HGSIL (P = 0.003), whereas no statistically significant difference was found between normal SIL and LGSIL (P = 0.1). Grouping observations by HPV status, no significant difference was detected in 8-OHdG levels between HPV(+) and HPV(-) subjects (P = 0.8). The polytomous and proportional odds models, extensions of the logistic regression analysis, showed that the effect of 8-OHdG levels in rising the risk of dysplasia was roughly constant through SIL grades. In conclusion, the immunoperoxidase method, applied to single human cervical cells, provides clear evidence that significant differences exist in 8-OHdG content between normal and dysplastic cells and that oxidative DNA damage might play an important role in cervical carcinogenesis.