P16/Ki-67 Immunostaining is Useful in Stratification of Atypical Metaplastic Epithelium of the Cervix

Cervical metaplastic squamous epithelium exhibiting atypia insufficient for a diagnosis of cervical intraepithelial neoplasia (CIN) is usually reported as “atypical squamous metaplasia” (ASM). Stratification impacts treatment since the differential is often between reactive and high grade CIN (CIN II, III). Diagnosis with H&E is associated with low intra/interobserver concurrence. P16/Ki-67 immunostains are helpful to assess cervical biopsies for HPV-associated lesions but staining in metaplastic squamous epithelium has received little attention. This study aims to establish staining characteristics of metaplastic squamous epithelium and determine if p16/Ki-67 is useful in ASM stratification. 80 cervical biopsies containing morphologically normal and dysplastic squamous metaplasia were retrieved to determine the staining characteristics of metaplastic epithelium utilizing p16/Ki-67 immunostains. These included 21 benign squamous metaplasia (BSM) from benign cervices, 15 BSM present adjacent to HPV/CIN lesions, and 44 CIN involving squamous metaplasia. Serial sections with controls were stained for p16 and Ki-67 and in-situ hybridization (ISH) for low-risk (LR) and high-risk (HR) HPV was performed. P16 was recorded as negative, spotty, or band-like. Ki-67 was recorded as positive when present in >50% of lesional nuclei. Results were correlated with H&E diagnosis. 95% of the BSMs, whether from normal cervices or adjacent to HPV/CIN were p16/Ki-67 negative. 81% HG CINs involving squamous metaplasia were p16 band/Ki-67 positive. Low grade CIN (CIN I) involving metaplastic epithelium showed a broad distribution of p16/Ki-67 staining patterns. Based on these criteria, 20 ASM were evaluated. 10% of the ASM cases were p16 band/Ki-67 positive indicating HG CIN. 60% of the ASMs were p16/Ki-67 negative indicating reactive change (all with the exception of one case being HPV negative). The remaining 30% of the ASM cases showed variable positivity for p16 and Ki-67 and could not be stratified into the two categories. Thus p16/Ki-67 staining is helpful in stratification of ASM as reactive or CIN

P16 and Ki67 Immunostains Decrease Intra- and Interobserver Variability in the Diagnosis and Grading of Anal Intraepithelial Neoplasia (AIN)

Background Significant variation is reported in the diagnosis of HPV-associated AIN. We previously observed that band-like positivity for p16 in >90% of contiguous cells coupled with Ki67 positivity in >50% of lesional cells is strongly associated with high grade AIN. This study was undertaken to determine if addition of p16 and Ki67 immunostaining would reduce inter- and intraobserver variability in diagnosis and grading of AIN. Design H&E stained slides of 60 anal biopsies were reviewed by three pathologists and consensus diagnoses were achieved: 25 negative, 12 low (condyloma and/or AIN I) and 23 high (9 AIN II and 14 AIN III) grade lesions. The H&E stained slides were diagnosed independently by three additional (“participant”) pathologists. Several weeks later they re-examined these slides in conjunction with corresponding p16 and Ki67 immunostains. Results Addition of p16 and Ki67 immunostains reduced intra- and interobserver variability, improved concurrence with consensus diagnoses and reduced two-step differences in diagnosis. Negative and high grade AIN diagnoses showed the most improvement in concurrence levels. Conclusion Addition of p16 and Ki67 immunostains is helpful in the diagnosis and grading of AIN

Overexpression of β1-chain-containing laminins in capillary basement membranes of human breast cancer and its metastases

Manabu Fujita, Natalya M Khazenzon, Shikha Bose, Kiyotoshi Sekiguchi, Takako Sasaki, William G Carter, Alexander V Ljubimov, Keith L Black and Julia Y Ljubimova, "Overexpression of β1-chain-containing laminins in capillary basement membranes of human breast cancer and its metastases", Breast Cancer Research, 7, R411-R421, BioMed Central, 200

WNT10B/β-catenin signalling induces HMGA2 and proliferation in metastatic triple-negative breast cancer

Wnt/β-catenin signalling has been suggested to be active in basal-like breast cancer. However, in highly aggressive metastatic triple-negative breast cancers (TNBC) the role of β-catenin and the underlying mechanism(s) for the aggressiveness of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally active β-catenin in human TNBC and predicts survival-outcome of patients with both TNBC and basal-like tumours. We provide evidence that transgenic murine Wnt10b-driven tumours are devoid of ERα, PR and HER2 expression and can model human TNBC. Importantly, HMGA2 is specifically expressed during early stages of embryonic mammogenesis and absent when WNT10B expression is lost, suggesting a developmentally conserved mode of action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical β-catenin signalling leading to up-regulation of HMGA2. Treatment of mouse and human triple-negative tumour cells with two Wnt/β-catenin pathway modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression predicts relapse-free-survival and metastasis in TNBC patients

RO Reject Water Management Techniques

In the last five decades, the variation for increase in population and decrease in available clean water is noticeable. To meet with growing water requirements, along with the wastewater from municipal sewage treatment plants, it is critical to consider the reuse of industrial wastewater wherever possible. In the Indian context, this has already been started in some of the big industries, especially in metro cities and it must be implemented at all level possible industries. Ultrafiltration (UF), reverse-osmosis (RO) and membrane bioreactors (MBR) will be integral parts of this. Waste water reuse is not only a necessity, it also provides several eco-friendly benefits