Application of CytoPath®Easy Vials in Cervical Cancer Screening: Self-Sampling Approach.

Context: CytoPath®Easy kit (DiaPath S.p.A.) offers a major advantage compared to other commercially available kits available for the screening of cervical cancer, as it does not require additional equipment for sample processing. Using this methodology, collected epithelial cells are immersed in a preservative liquid before setting as a thin layer on a slide via gravity sedimentation. Aims: To evaluate the suitability of the CytoPath®Easy kit for the processing of cervical samples, detection of pre-neoplastic lesions, and nucleic preservation and extraction for HR-HPV diagnosis. Materials and Methods: A total of 242 self-sampled cervical specimens were utilized, with 192 collected in CytoPath®Easy vials and 50 collected and processed using the ThinPrepTM for comparative analysis. The samples underwent processing, Papanicolaou staining, and microscopic evaluation for morphological parameters. The extracted nucleic acids were assessed for purity and integrity, and the detection of high-risk human papillomavirus (HR-HPV) was carried out using the Alinitym HR HPV system kit (Abbott Laboratórios Lda). Results: Both methods demonstrated effective performance, enabling the morphological assessment of the cervical epithelium. Statistical analysis indicated that ThinPrepTM yielded significantly better results in terms of cellularity. Conversely, CytoPath®Easy exhibited superior performance in terms of the quantity of extracted DNA and its degree of purification. Concerning the time consumed during processing, both methods were comparable, with the CytoPath®Easy methodology standing out for its cost-effectiveness, as it does not necessitate additional instruments and consumables. Conclusions: The novel CytoPath®Easy methodology proves effective in preserving both nucleic acids and cell morphology characteristics, two crucial features for cervical cancer screening.

Human Epidermal Growth Factor Receptor 2 (HER2) Expression by Immunohistochemistry and Its Clinical Significance in Hepatocellular Carcinoma: A Single-Center Analysis.

BACKGROUND: Human epidermal growth factor receptor 2 (HER2) is a member of the tyrosine kinase receptor family. It has been identified as an oncogene and is associated with poor outcomes in multiple tumor types. In hepatocellular carcinoma (HCC), there are contradictory data regarding the HER2 expression and its role in tumor development and progression. Some studies have identified HER2 expression as an early event during tumorigenesis, which decreases with progression and metastasis. Additional data provided evidence that treatment with anti-HER2 therapy resulted in local response and reduction in the metastasis rate in HCC mice models. METHODS: Patients with histological diagnoses of HCC between 2010 and 2020 were included. HER2 staining was performed by immunohistochemistry (IHC), and scoring was done in accordance with the gastric cancer guidelines as 0, 1+, 2+, and 3+. Clinicopathological features were accessed by medical records. This study aims to evaluate HER2 expression by IHC in HCC and to correlate this expression with some clinicopathological features such as Barcelona Clinic Liver Cancer (BCLC) staging, number of hepatic lesions, alpha-fetoprotein level, underlying liver disease, presence of liver cirrhosis, Child-Pugh score, and tumor recurrence. RESULTS: A total of 57 specimens from 54 patients were included. Of the patients, 85% were men, and the median age at diagnosis was 71 years (interquartile range: 59-75 years). Regarding stage, 61% were at stage 0-A of BCLC. Of the patients, 57% had a solitary HCC nodule. Concerning treatment, surgery was performed in 50% of the patients. HER2 expression was identified in seven patients: five in the membrane and two in the cytoplasm. Concerning the membrane staining, HER2 expression was scored as 1+/2+ in 7.4% (n = 4 patients). Of the patients with HER2 expression, four had a BCLC stage of 0-A and a single HCC nodule; alpha-fetoprotein was <400 ng/mL in all cases. There was no correlation to clinicopathological features. In one patient with HER2 2+ expression at diagnosis, this expression was not identified at tumor progression. Median disease-free survival in HER2 with IHC scores 1+/2+ and cytoplasmatic was 38 months versus 22 months in HER2 with a score of 0 (p = 0.604). CONCLUSIONS: HER2 expression is a rare event in HCC. It was not possible to identify any relation to clinicopathological features. However, when we relate our data to previous trials, HER2 appears to be an early event in the course of HCC.

Molecular Staging of Patients with Colon Cancer. The C-Closer-II Study: A Multicentre Study in Portugal.

INTRODUCTION: Approximately 20% - 30% of histological lymph node-negative patients with colorectal cancer relapse at five years after surgical treatment. This recurrence is likely due to occult nodal disease undetected by standard histopathological practice which has implications in terms of the clinical management of patients. MATERIAL AND METHODS: Lymph nodes were collected from colectomy specimens. A central section from each lymph node was histologically examined following haematoxylin-eosin staining and the remaining tissue was subjected to OSNA - one step nucleic acid amplification analysis. RESULTS: A total of 1046 lymph nodes from 59 pN0 patients were assessed. Of these, 753 lymph nodes were examined by both methods. The median number of lymph nodes assessed with OSNA - one step nucleic acid amplification was 12 (IQR: 7;16). Among pN0 patients, 17 had OSNA - one step nucleic acid amplification-positive lymph nodes, resulting in a positive molecular staging rate of 28.8% (95% CI: 17.8 - 42.1). Among these patients, 12 (70.59%) were molecular-staged as pN1 and 5 (29.41%) were molecular staged as pN2. The tumour burden of lymph nodes assessed with OSNA - one step nucleic acid amplification ranged from 270 to 17 000 cytokeratin 19 mRNA copies/µL. Most of these patients (88.2%) were found to have lymph nodes with micrometastases only (250 - 4999 copies/µL). DISCUSSION: We provide the results from the first study of the use of the OSNA - one step nucleic acid amplification assay in colorectal cancer patients in Portugal. Our results are in-line with other international studies, showing the improvement on patients' staging by molecular examination of lymph nodes. CONCLUSION: In our study, 28.8% of patients with histologically negative lymph nodes were found to have metastatic lymph nodes using OSNA - one step nucleic acid molecular assessment. OSNA - one step nucleic acid assay allows a more accurate staging of patients with colorectal cancer and standardizes lymph node assessment.

Introdução: Cerca de 20% - 30% dos doentes com cancro colo-rectal, com gânglios linfáticos regionais negativos por histologia têm recidiva do carcinoma colo-rectal, após cinco anos do tratamento cirúrgico. Esta recorrência é provavelmente devida à presença de metástases ganglionares ocultas, não detetadas no exame anatomo-patológico standard. Material e Métodos: Os gânglios linfáticos foram obtidos a partir de espécimes de colectomia. Uma secção central de cada gânglio linfático foi analisada histologicamente com a coloração de hematoxilina-eosina e o tecido restante foi sujeito a análise de OSNA - one step nucleic acid amplification. Resultados: Um total de 1046 gânglios linfáticos de 59 doentes pN0 foram avaliados. Foram examinados 753 gânglios linfáticos por ambos os métodos. A mediana de gânglios linfáticos avaliados com OSNA - one step nucleic acid amplification foi de 12 (IQR:7; 16). Entre os doentes pN0, 17 tinham gânglios linfáticos positivos para OSNA - one step nucleic acid amplification, resultando numa taxa de estadiamento molecular positiva de 28,8% (95% CI: 17,8 - 42,1). De entre esses doentes, 12 (70,59%) apresentaram-se molecularmente pN1 e cinco (29,41%) pN2. A carga tumoral dos gânglios linfáticos avaliada com OSNA - one step nucleic acid amplification variou de 270 a 17 000 cópias/µL de ARNm de citoqueratina 19. A maioria desses doentes (88,2%) apresentou gânglios linfáticos com micrometástases (250 - 4999 cópias/µL). Discussão: Apresentamos os resultados do primeiro estudo do ensaio OSNA - one step nucleic acid amplification levado a cabo, em pacientes com cancro colo-rectal, em Portugal. Os nossos resultados estão em linha com outros estudos internacionais, demostrando uma melhoria no estadiamento dos doentes, pelo exame molecular dos gânglios linfáticos. Conclusão: Verificou-se que 28,8% dos doentes com gânglios linfáticos histologicamente negativos apresentavam doença ganglionar metastática, usando a avaliação molecular de OSNA - one step nucleic acid amplification. O ensaio OSNA - one step nucleic acid amplification permite um estadiamento mais preciso de doentes com carcinoma do colon e padroniza a avaliação dos gânglios linfáticos.

Structural and molecular analysis of the cancer prostate cell line PC3: Oocyte zona pellucida glycoproteins.

The human oocyte zona pellucida (ZP) is made of four glycoproteins, ZP1-ZP4. Recently, the prostate adenocarcinoma and prostate cancer PC3 cell-line were shown to express the human oocyte ZP3 glycoprotein, which was evaluated in a single report subject to patent. To further clarify whether oocyte zona pellucida glycoproteins are expressed in prostate cancer tissue and PC3-cells, in this report we evaluated protein expression of the four ZP glycoproteins in normal prostate tissue, prostate adenocarcinoma tissue and PC3-cells, and performed quantitative mRNA expression of the four ZP glycoproteins in the PC3 cell-line. Furthermore, as PC3-cells have not yet been studied in detail regarding their ultrastructural characteristics, in the present report we bring forward the detailed ultrastructure of PC3-cells. PC3-cells were divided into pavement and aggregated cells. We observed new ultrastructural features in pavement and aggregated cells, with the later exhibiting two different cell types. In prostate carcinoma tissue and PC3-cells we found protein expression of the four oocyte glycoproteins, ZP1, ZP2, ZP3 and ZP4. In addition, mRNA expression studies revealed expression of ZP1, ZP3 and ZP4 glycoproteins, but not of ZP2. Interestingly, the ZP1 mRNA product exhibited intron retention.