Molecular analysis and application of tissue microarray technology to the histopathological and immunohistochemical analysis of cervical adenocarcinoma

Cervical cancer is the second most common cancer among women worldwide. Cervical adenocarcinoma accounts for 15-25% of all cervical cancers and the incidence appears to be increasing. Although some of this increase may be due to better recognition by pathologists, there is evidence that the incidence of both endocervical adenocarcinoma and its preinvasive precursor lesion cervical glandular intraepithelial neoplasia is actually increasing in real terms. In this study, tissue microarray technology was used to study the morphological features of cervical adenocarcinoma archival donor blocks, to evaluate the immunoprofile of a large set of cervical adenocarcinomas with an extended panel of antibodies to compare the profile of AIS with invasive subtypes of cervical adenocarcinomas. The prevalence of HPV 16&18 in cervical adenocarcinoma cases was assessed to evaluate its relation to cervical adenocarcinoma. Using haematoxylin and eosin staining method 273 samples (blocks) were obtained from 177 biopsies composed of 16 normal cervical biopsies, 139 different patients with endocervical adenocarcinomas, and 22 patients with second biopsies. Pathology reports and cervical smear history reports were reviewed. Morphological and histopathological features of 139 patients with cervical adenocarcinomas revealed that there were 20 patients with adenocarcinoma-in-situ and 119 with invasive adenocarcinoma. Sixteen of 119 patients with invasive adenocarcinoma had early invasive adenocarcinoma which met criteria for FIGO stage IAi carcinoma of the cervix. The tissue microarray technique has been demonstrated to be efficient and applicable to various tumour types, but methodological evaluations are few. A tissue microarray was constructed using paraffin-embedded, formalin-fixed tissues from 273 samples (blocks). Two paired 0.6-mm cores were obtained from selected regions of archival donor blocks and subsequently were arrayed into a recipient paraffin array blocks. More than 2 areas were taken from some tumours. The novel array blocks and some whole sections were used for immunohistochemical analysis and H&E staining. Results revealed that the tissue microarray method yields staining of good quality and is feasible for histopathological and immunohistochemical studies in cervical adenocarcinoma. In general, the average staining pattern agreed with the whole section in each. Analysis of two TMA cores achieved 100% representation for histopathological type and greater than 97% representation for immunohistochemical studies. Tissue array sections were immunostained with 8 antibodies, carcinoembryonic antigen (CEA), Cytokeratin7 (CK7), Cytokeratin20 (CK20), estrogen receptor (ER), progesterone receptor (PgR), phosphatase and tensin homolog deleted on chromosome ten (PTEN), MIB-1 proliferation marker, and p53 suppressor gene utilizing the "Power Vision" technique for ER only and "Envision" technique for all other antibodies. Our findings support that all of these 8 antibodies are of potential biomarkers of a panel of antibodies for diagnosis of cervical adenocarcinomas. HPV DNA was extracted from paraffin-embedded, formalin-fixed tissues of 161 specimens of 139 patients excluding 22 patients with second samples and 16 normal cervical tissues. HPV DNA was detected by PCR test using type specific primers from the E6 gene and E7 gene of HPV type 16 and HPV type 18. Out of a total of 257 cervical biopsies from 139 women with various cervical adenocarcinomas lesions, HPV DNA was identified in 87 cases (62.6%) in which, HPV16 was positive for 65 (47%) patients and HPV 18 was positive for 41 (29%) patients. Genotyping by RFLP and PCR revealed that HPV type 16 was the most frequent type of infection comprising 46 cases (33%), followed by HPV type 18 in 22 cases (16%), and both HPV typel6 and HPV type 18 in 19 cases (14%). HPV typing in all cases of 16 normal cervical biopsies was negative with both HPV typel6 and HPV type 18. The findings support that HPV 16, along with HPV 18, may play a possible role in the pathogenesis of adenocarcinoma of the uterine cervix

Validation of tissue microarray technology in squamous cell carcinoma of the esophagus

Tissue microarray (TMA) technology has been developed to facilitate high-throughput immunohistochemical and in situ hybridization analysis of tissues by inserting small tissue biopsy cores into a single paraffin block. Several studies have revealed novel prognostic biomarkers in esophageal squamous cell carcinoma (ESCC) by means of TMA technology, although this technique has not yet been validated for these tumors. Because representativeness of the donor tissue cores may be a disadvantage compared to full sections, the aim of this study was to assess if TMA technology provides representative immunohistochemical results in ESCC. A TMA was constructed containing triplicate cores of 108 formalin-fixed, paraffin-embedded squamous cell carcinomas of the esophagus. The agreement in the differentiation grade and immunohistochemical staining scores of CK5/6, CK14, E-cadherin, Ki-67, and p53 between TMA cores and a subset of 64 randomly selected donor paraffin blocks was determined using kappa statistics. The concurrence between TMA cores and donor blocks was moderate for Ki-67 (κ = 0.42) and E-cadherin (κ = 0.47), substantial for differentiation grade (κ = 0.65) and CK14 (κ = 0.71), and almost perfect for p53 (κ = 0.86) and CK5/6 (κ = 0.93). TMA technology appears to be a valid method for immunohistochemical analysis of molecular markers in ESCC provided that the staining pattern in the tumor is homogeneous

SARS-CoV-2 vaccination modelling for safe surgery to save lives: data from an international prospective cohort study

Background: Preoperative SARS-CoV-2 vaccination could support safer elective surgery. Vaccine numbers are limited so this study aimed to inform their prioritization by modelling. Methods: The primary outcome was the number needed to vaccinate (NNV) to prevent one COVID-19-related death in 1 year. NNVs were based on postoperative SARS-CoV-2 rates and mortality in an international cohort study (surgical patients), and community SARS-CoV-2 incidence and case fatality data (general population). NNV estimates were stratified by age (18-49, 50-69, 70 or more years) and type of surgery. Best- and worst-case scenarios were used to describe uncertainty. Results: NNVs were more favourable in surgical patients than the general population. The most favourable NNVs were in patients aged 70 years or more needing cancer surgery (351; best case 196, worst case 816) or non-cancer surgery (733; best case 407, worst case 1664). Both exceeded the NNV in the general population (1840; best case 1196, worst case 3066). NNVs for surgical patients remained favourable at a range of SARS-CoV-2 incidence rates in sensitivity analysis modelling. Globally, prioritizing preoperative vaccination of patients needing elective surgery ahead of the general population could prevent an additional 58 687 (best case 115 007, worst case 20 177) COVID-19-related deaths in 1 year. Conclusion: As global roll out of SARS-CoV-2 vaccination proceeds, patients needing elective surgery should be prioritized ahead of the general population