Metasin: An Intra-Operative Real-Time Quantitative Reverse Transcription Polymerase-Chain Reaction (RTqPCR) Assay to Detect Metastatic Breast Cancer in Sentinel Lymph Nodes

The copyright of this thesis rests with the author and no quotation from it or information derived from it may be published without the prior written consent of the authorThe most important prognostic factor in breast cancer is the presence or absence of metastases in axillary lymph nodes. Frozen section and touch imprint cytology are conventional intra-operative methods used in the detection of metastatic breast cancer with varying sensitivities and specificities. The limitation of these methods led to the development of alternative molecular diagnostic tests, such as GeneSearch, a commercial real-time quantitative Polymerase Chain Reaction (RT-qPCR) assay that allows for an intra-operative diagnosis of metastatic breast cancer. When the GeneSearch assay was discontinued, Metasin was developed as an in-house RT-qPCR replacement assay. Metasin targets the epithelial cell marker cytokeratin 19 (CK19) and the breast marker mammaglobin (MGB) mRNA to confirm the presence or absence of metastatic disease, whilst the reference gene porphobilinogen deaminase (PBGD) acts as a positive control for the performance of the assay. The optimised assay can produce a result within 32 minutes allowing it to be used in the intra-operative setting to detect metastatic breast cancer in sentinel lymph nodes. 154 archived lymph node homogenates that were previously analysed by both GeneSearch and histology in parallel were used to validate Metasin. Out of 154 cases, 148 showed concordance with both GeneSearch and Metasin with 111 cases being negative and 37 cases being positive. There were six discordant cases, four in which only Metasin detected metastases and two in which only GeneSearch picked up metastases. Out of the four Metasin-only positive cases, three were found to be positive on histology after deeper levels were cut in the slices sent for histological assessment. Therefore, one case could not be shown histologically to be positive for metastases. There were two cases that were missed by Metasin but picked up by GeneSearch. One case was positive on histology and the second case negative for histology. The error rate for Metasin was 3.89%. The sensitivity and specificity of the Metasin assay were found to be 95% and 98% respectively, and the positive and negative predictive values were 90% and 98% respectively. These results are comparable to those of GeneSearch. Metasin had an assay time of less than 45 minutes and was operated by biomedical scientists. The results of the validation process were deemed acceptable for the assay to be run live and used in the clinical setting. Metasin continues to provide breast cancer patients at Princess Alexandra Hospital with all the advantages that a molecular intra-operative diagnostic service provides.Epping Breast Cancer Research Fun

NILAI FILOSOFIS PROSES KHITBAH DAN MENENTUKAN MAHAR PERKAWINAN UNTUK PEREMPUAN DALAM TINJAUAN FEMINISME ISLAM (TAFSIR AL-BAQARAH : 235-237 DAN AN-NISAA' : 4)

The dowry is a gift made by the bridegroom to the bride with an obligatory law but it's not mentioned in the form and type, large and small both in the Qur'an and in the hadith and khitbah as a statement made by a man to a woman by expressing a desire to marry a woman as her life partner. This is seen in the perspective of Islamic feminism by interpreting the verses of the Qur'an to obtain the meaning of gender justice, this is because the interpretation that has been done traditionally is sometimes still gender biased and makes men superior. The method used is a descriptive qualitative method with research that explains the reality of the understanding of a social phenomenon that occurs in society and discussed in more detail. This study states that long before there was an issue of gender quality, the verses of the Qur'an glorified women, had long before paid attention to the position of women with the interpretation of Q.S An-Nisaa' : 4 about giving dowry Q.S. Al-Baqarah : 235 about the khitbah process and Q.S. Al-Baqarah : 236-237 about Mut'ah given to women who were given away

A multicentre validation of Metasin: a molecular assay for the intraoperative assessment of sentinel lymph nodes from breast cancer patients

Aims: Treatment strategies for breast cancer continue to evolve. No uniformity exists in the UK for the management of node‐positive breast cancer patients. Most centres continue to use conventional histopathology of sampled sentinel lymph nodes (SLNs), which requires delayed axillary clearance in up to 25% of patients. Some use touch imprint cytology or frozen section for intraoperative testing, although both have inherent sensitivity issues. An intraoperative molecular diagnostic approach helps to overcome some of these limitations. The aim of this study was to assess the clinical effectiveness of Metasin, a molecular method for the intraoperative evaluation of SLNs. Methods and results: RNA from 3296 lymph nodes from 1836 patients undergoing SLN assessment was analysed with Metasin. Alternate slices of tissue were examined in parallel by histology. Cases deemed to be discordant were analysed by protein gel electrophoresis. There was concordance between Metasin and histology in 94.1% of cases, with a sensitivity of 92% [95% confidence interval (CI) 88–94%] and a specificity of 97% (95% CI 95–97%). Positive and negative predictive values were 88% and 98%, respectively. Over half of the discordant cases (4.4%) were ascribed to tissue allocation bias (TAB). Conclusions: Clinical validation of the Metasin assay suggests that it is sufficiently sensitive and specific to make it fit for purpose in the intraoperative setting

Metasin — An Intra-Operative RT-qPCR Assay to Detect Metastatic Breast Cancer in Sentinel Lymph Nodes

Nodal status is one of the most important prognostic factors in breast cancer. Established tests such as touch imprint cytology and frozen sections currently used in the intra-operative setting show variations in sensitivity and specificity. This limitation has led to the development of molecular alternatives, such as GeneSearch, a commercial intra-operative real-time quantitative Polymerase Chain Reaction (RT-qPCR) assay that allows the surgeon to carry out axillary clearance as a one-step process. Since GeneSearch has been discontinued, we have developed the replacement Metasin assay, which targets the breast epithelial cell markers CK19 and mammaglobin mRNA and identifies metastatic disease in sentinel lymph nodes. The optimised assay can be completed within 32 min (6 min for RNA preparation and 26 min instrument run time), making its use feasible in the intraoperative setting. An analysis by Metasin of 154 archived lymph node homogenates previously analysed by both parallel histology and GeneSearch showed concordance for 148 cases. The sensitivity and specificity of Metasin compared with GeneSearch were 95% (CI 83%-99%) and 97% (CI 91%-99%) respectively; compared with histology they were 95% (CI 83%-99%) and 97% (CI 91%-99%), respectively. The sensitivity and specificity of GeneSearch compared with histology were 90% (CI 77%-96%) and 97% (CI 93%-99%) respectively. The positive predictive value of Metasin was 90% and negative predictive value was 98% for both histology and GeneSearch. The positive predictive value of GeneSearch was 92% and the negative predictive value was 97% compared to histology. The discordance rates of Metasin with both GeneSearch and histology were 3.89%. In comparison, the discordance rate of GeneSearch with histology was 4.5%. Metasin's robustness was independently evaluated on 193 samples previously analysed by GeneSearch from the Jules Bordet Institute, where Metasin yielded comparable results

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

SummaryBackground Azithromycin has been proposed as a treatment for COVID-19 on the basis of its immunomodulatoryactions. We aimed to evaluate the safety and efficacy of azithromycin in patients admitted to hospital with COVID-19.Methods In this randomised, controlled, open-label, adaptive platform trial (Randomised Evaluation of COVID-19Therapy [RECOVERY]), several possible treatments were compared with usual care in patients admitted to hospitalwith COVID-19 in the UK. The trial is underway at 176 hospitals in the UK. Eligible and consenting patients wererandomly allocated to either usual standard of care alone or usual standard of care plus azithromycin 500 mg once perday by mouth or intravenously for 10 days or until discharge (or allocation to one of the other RECOVERY treatmentgroups). Patients were assigned via web-based simple (unstratified) randomisation with allocation concealment andwere twice as likely to be randomly assigned to usual care than to any of the active treatment groups. Participants andlocal study staff were not masked to the allocated treatment, but all others involved in the trial were masked to theoutcome data during the trial. The primary outcome was 28-day all-cause mortality, assessed in the intention-to-treatpopulation. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936.Findings Between April 7 and Nov 27, 2020, of 16 442 patients enrolled in the RECOVERY trial, 9433 (57%) wereeligible and 7763 were included in the assessment of azithromycin. The mean age of these study participants was65·3 years (SD 15·7) and approximately a third were women (2944 [38%] of 7763). 2582 patients were randomlyallocated to receive azithromycin and 5181 patients were randomly allocated to usual care alone. Overall,561 (22%) patients allocated to azithromycin and 1162 (22%) patients allocated to usual care died within 28 days(rate ratio 0·97, 95% CI 0·87–1·07; p=0·50). No significant difference was seen in duration of hospital stay (median10 days [IQR 5 to >28] vs 11 days [5 to >28]) or the proportion of patients discharged from hospital alive within 28 days(rate ratio 1·04, 95% CI 0·98–1·10; p=0·19). Among those not on invasive mechanical ventilation at baseline, nosignificant difference was seen in the proportion meeting the composite endpoint of invasive mechanical ventilationor death (risk ratio 0·95, 95% CI 0·87–1·03; p=0·24).Interpretation In patients admitted to hospital with COVID-19, azithromycin did not improve survival or otherprespecified clinical outcomes. Azithromycin use in patients admitted to hospital with COVID-19 should be restrictedto patients in whom there is a clear antimicrobial indication