Artificial Intelligence to Predict the BRAF V595E Mutation in Canine Urinary Bladder Urothelial Carcinomas

In dogs, the BRAF mutation (V595E) is common in bladder and prostate cancer and represents a specific diagnostic marker. Recent advantages in artificial intelligence (AI) offer new opportunities in the field of tumour marker detection. While AI histology studies have been conducted in humans to detect BRAF mutation in cancer, comparable studies in animals are lacking. In this study, we used commercially available AI histology software to predict BRAF mutation in whole slide images (WSI) of bladder urothelial carcinomas (UC) stained with haematoxylin and eosin (HE), based on a training (n = 81) and a validation set (n = 96). Among 96 WSI, 57 showed identical PCR and AI-based BRAF predictions, resulting in a sensitivity of 58% and a specificity of 63%. The sensitivity increased substantially to 89% when excluding small or poor-quality tissue sections. Test reliability depended on tumour differentiation (p < 0.01), presence of inflammation (p < 0.01), slide quality (p < 0.02) and sample size (p < 0.02). Based on a small subset of cases with available adjacent non-neoplastic urothelium, AI was able to distinguish malignant from benign epithelium. This is the first study to demonstrate the use of AI histology to predict BRAF mutation status in canine UC. Despite certain limitations, the results highlight the potential of AI in predicting molecular alterations in routine tissue sections

Artificial Intelligence to Predict the BRAF V595E Mutation in Canine Urinary Bladder Urothelial Carcinomas.

In dogs, the BRAF mutation (V595E) is common in bladder and prostate cancer and represents a specific diagnostic marker. Recent advantages in artificial intelligence (AI) offer new opportunities in the field of tumour marker detection. While AI histology studies have been conducted in humans to detect BRAF mutation in cancer, comparable studies in animals are lacking. In this study, we used commercially available AI histology software to predict BRAF mutation in whole slide images (WSI) of bladder urothelial carcinomas (UC) stained with haematoxylin and eosin (HE), based on a training (n = 81) and a validation set (n = 96). Among 96 WSI, 57 showed identical PCR and AI-based BRAF predictions, resulting in a sensitivity of 58% and a specificity of 63%. The sensitivity increased substantially to 89% when excluding small or poor-quality tissue sections. Test reliability depended on tumour differentiation (p < 0.01), presence of inflammation (p < 0.01), slide quality (p < 0.02) and sample size (p < 0.02). Based on a small subset of cases with available adjacent non-neoplastic urothelium, AI was able to distinguish malignant from benign epithelium. This is the first study to demonstrate the use of AI histology to predict BRAF mutation status in canine UC. Despite certain limitations, the results highlight the potential of AI in predicting molecular alterations in routine tissue sections

Status of the BELLE II Pixel Detector

The Belle II experiment at the super KEK B-factory (SuperKEKB) in Tsukuba, Japan, has been collecting $e^+e^−$ collision data since March 2019. Operating at a record-breaking luminosity of up to $4.7×10^{34} cm^{−2}s^{−1}$, data corresponding to $424 fb^{−1}$ has since been recorded. The Belle II VerteX Detector (VXD) is central to the Belle II detector and its physics program and plays a crucial role in reconstructing precise primary and decay vertices. It consists of the outer 4-layer Silicon Vertex Detector (SVD) using double sided silicon strips and the inner two-layer PiXel Detector (PXD) based on the Depleted P-channel Field Effect Transistor (DePFET) technology. The PXD DePFET structure combines signal generation and amplification within pixels with a minimum pitch of $(50×55) μm^2$. A high gain and a high signal-to-noise ratio allow thinning the pixels to $75 μm$ while retaining a high pixel hit efficiency of about $99$. As a consequence, also the material budget of the full detector is kept low at $≈0.21$$\frac{X}{X_0}$ per layer in the acceptance region. This also includes contributions from the control, Analog-to-Digital Converter (ADC), and data processing Application Specific Integrated Circuits (ASICs) as well as from cooling and support structures. This article will present the experience gained from four years of operating PXD; the first full scale detector employing the DePFET technology in High Energy Physics. Overall, the PXD has met the expectations. Operating in the intense SuperKEKB environment poses many challenges that will also be discussed. The current PXD system remains incomplete with only 20 out of 40 modules having been installed. A full replacement has been constructed and is currently in its final testing stage before it will be installed into Belle II during the ongoing long shutdown that will last throughout 2023

Perinatal and 2-year neurodevelopmental outcome in late preterm fetal compromise: the TRUFFLE 2 randomised trial protocol

Introduction: Following the detection of fetal growth restriction, there is no consensus about the criteria that should trigger delivery in the late preterm period. The consequences of inappropriate early or late delivery are potentially important yet practice varies widely around the world, with abnormal findings from fetal heart rate monitoring invariably leading to delivery. Indices derived from fetal cerebral Doppler examination may guide such decisions although there are few studies in this area. We propose a randomised, controlled trial to establish the optimum method of timing delivery between 32 weeks and 36 weeks 6 days of gestation. We hypothesise that delivery on evidence of cerebral blood flow redistribution reduces a composite of perinatal poor outcome, death and short-term hypoxia-related morbidity, with no worsening of neurodevelopmental outcome at 2 years. Methods and analysis: Women with non-anomalous singleton pregnancies 32+0 to 36+6 weeks of gestation in whom the estimated fetal weight or abdominal circumference is &lt;10th percentile or has decreased by 50 percentiles since 18-32 weeks will be included for observational data collection. Participants will be randomised if cerebral blood flow redistribution is identified, based on umbilical to middle cerebral artery pulsatility index ratio values. Computerised cardiotocography (cCTG) must show normal fetal heart rate short term variation (≥4.5 msec) and absence of decelerations at randomisation. Randomisation will be 1:1 to immediate delivery or delayed delivery (based on cCTG abnormalities or other worsening fetal condition). The primary outcome is poor condition at birth and/or fetal or neonatal death and/or major neonatal morbidity, the secondary non-inferiority outcome is 2-year infant general health and neurodevelopmental outcome based on the Parent Report of Children's Abilities-Revised questionnaire. Ethics and dissemination: The Study Coordination Centre has obtained approval from London-Riverside Research Ethics Committee (REC) and Health Regulatory Authority (HRA). Publication will be in line with NIHR Open Access policy. Trial registration number: Main sponsor: Imperial College London, Reference: 19QC5491. Funders: NIHR HTA, Reference: 127 976. Study coordination centre: Imperial College Healthcare NHS Trust, Du Cane Road, London, W12 0HS with Centre for Trials Research, College of Biomedical &amp; Life Sciences, Cardiff University. IRAS Project ID: 266 400. REC reference: 20/LO/0031. ISRCTN registry: 76 016 200

Double-balloon catheter for induction of labour in women with a previous cesarean section, could it be the best choice?

INTRODUCTION: We analysed the efficacy and safety of double-balloon catheter for cervical ripening in women with a previous cesarean section and which were the most important variables associated with an increased risk of repeated cesarean delivery. MATERIALS AND METHODS: We designed an observational retrospective study of 418 women with unfavourable cervices (Bishop Score <5), a prior cesarean delivery, and induction of labour with a double-balloon catheter. Baseline maternal data and perinatal outcomes were recorded for a descriptive, bivariate, and multivariate analysis. A p value <0.05 was considered statistically significant. RESULTS: Most women improved their initial Bishop Score (89.5%) although only a 20.8% of them went into spontaneous active labour. Finally, 51.4% of the women achieved a vaginal delivery. Five cases of intrapartum uterine rupture (1.2%) occurred. After multivariate analysis, main risk factors for repeated cesarean section were dystocia in the previous pregnancy (OR 1.744; CI 95% 1.066–2.846), the absence of previous vaginal delivery (OR 2.590; CI 95% 1.066–6.290), suspected fetal macrosomia (OR 2.410; CI 95% 0.959–6.054), and duration of oxytocin induction period (OR 1.005; CI 95% 1.004–1.006). The area under the curve was 0.789 (p < 0.001). CONCLUSIONS: Double-balloon catheter seems to be safe and effective for cervical ripening in women with a previous cesarean delivery and unfavourable cervix. In our study, most women could have a vaginal delivery in spite of their risk factors for cesarean delivery. A multivariate model based on some clinical variables has moderate predictive value for intrapartum cesarean section

Cytologic intraoperative diagnosis of breast cancer

Zytologische Schnelldiagnose des Mammakarzinoms Einleitung Die intraoperative Befundung eines suspekten Mammabefundes wird durch die histologische Aufarbeitung des zu untersuchenden Präparates erreicht. Da jede Methode seine Grenzen hat und heutzutage eine für den Patienten optimalste Behandlung angestrebt wird, stellte sich die Frage, ob man die histologische Abklärung durch eine weitere Methode sinnvoll ergänzen kann. Unter diesem Gesichtspunkt wurde bei 111 Patientinnen zusätzlich zur histologischen Schnellschnittuntersuchung eine Imprintzytologie, die mit einer Schnellfärbung behandelt wurde, durchgeführt. Das Patientenkollektiv entspricht dem Patientengut einer universitären Frauenklinik mit Mammazentrum und hat seinen einzigsten gemeinsamen Nenner in der Notwendigkeit einer operativen Abklärung eines Mammabefundes. Herstellung des Imprintes Während der operativen Abklärung eines Mammabefundes wird das zu untersuchende Präparat, als Schnellschnitt deklariert, in die Pathologie gebracht. Der Pathologe versucht nun Aussagen über die Dignität des Präparates und die Tumorzellfreiheit der Resektionsränder zu treffen. Hierzu öffnet er einerseits das Tumorzentrum und setzt weiterhin Schnitte an den Resektionsrändern. Je nach Beschaffenheit des Präparates kann nun durch sanftes Drücken oder durch Streifen des Tumorzentrums auf einen Objektträger ein Imprint gewonnen werden. Ziel dieses Vorganges ist das Erhalten einer ausreichend repräsentativen Menge an Tumorzellen, die eine anschließende zytologische Beurteilung gewährleisten soll. Um die Sicherheit der zytologischen Diagnose zu erhöhen, werden zwei Imprints angefertigt. Nun kann die histologische Befundung verzögerungsfrei erfolgen, die parallel durch die Schnellfärbung und zytologische Beurteilung des Imprintes ergänzt wird. Schnellfärbung des Imprintes Im Vergleich zu konventionellen Färbemethoden, der MGG ( May-Grünwald-Giemsa )- Färbung oder der Färbemethode nach Papanicolaou, die einen längeren Zeitraum zur Ausbildung ihres vollen Färbeeffektes benötigen, ermöglicht die Schnellfärbung eine Beurteilung innerhalb weniger Minuten. Der Imprint wird hierbei mit einer bereiteten Lösung , einem Gemisch aus MGG und Methanol, bedeckt und nach zwei Minuten mit Aqua dest. sorgfältig abgespült. Daraufhin wird es mit einem zweiten Gemisch aus Giemsa und Aqua dest. wiederum für zwei Minuten behandelt. Nach erfolgter Abspülung mit Aqua dest. kann die Beurteilung des Imprintes durch den Zytologen erfolgen. Hierfür werden lediglich zytologische Kriterien herangezogen, da weitere Informationen über den Patienten und des Präparates fehlen. Nach retrospektivem Vergleich der zytolgischen Schnelldiagnosen mit dem endgültigen histologischen Befund, die eine Schwäche bei der Erkennung von Fibroadenomen offenbarte, wurde eine primäre Färbung mit Mayers- Hämalaun eingeführt. Nachdem der Imprint für wenige Sekunden mit Mayers- Hämalaun gefärbt wurde, folgt der oben beschriebene Ablauf. Ergebnisse Die vom Zytologen gestellte Diagnose wird, nachdem sie im eigens kreierten Datenblatt vermerkt wurde, mit der histologischen Schnellschnittdiagnose und der endgültigen histologischen Diagnose verglichen. Die 111 untersuchten Präparate zeigten in 93 Fällen eine maligne und in 18 Fällen eine gutartige Veränderung. Die zytologische Schnelldiagnose erkannte 90 der 93 malignen Geschehen und 14 der benignen Veränderungen. Neben zwei falsch positiven und einer falsch- negativen Diagnose konnten vier Präparate aufgrund ungenügenden Zellmaterials nicht beurteilt werden. Die Sensitivität der zytologischen Untersuchung beträgt somit 97% und die Spezifität 78%. Die histologische Schnellschnittdiagnose erkannte 90 der 93 malignen Veränderungen und alle 18 positiven Veränderungen. Dies entspricht einer Sensitivität von 97% und einer Spezifität von 100%. Bemerkungen Bei der zytologischen Schnelldiagnose sind vor allem zwei Aspekte zu beachten. Erstens ist die Imprintgewinnung für die exakte Diagnosenstellung von elementarer Bedeutung, denn hier gilt es, ausreichend repräsentatives Zellmaterial zu gewinnen. Zweitens muss man sich der möglichen Fehlerquellen einer zytologischen Begutachtung bewusst sein. Das Fibroadenom kann beispielsweise leicht als ein maligner Prozess missinterpretiert werden. Dies war für die falsch- positiven Diagnosen verantwortlich. Mit der Einführung einer primären Mayers- Hämalaun Färbung wird versucht, über die Darstellung von Myoepithelien, die Erkennbarkeit des Fibroadenoms zu erleichtern. Schlussfolgerung Die zusätzliche zytologische Schnelluntersuchung zur histologischen Schnellschnittuntersuchung ist von Vorteil. Die zytologische Schnelldiagnostik hat eine hohe Sensitivität, die von der Qualität der Abnahmebedingungen abhängt. Die Spezifität der zytologischen Methode und dabei vor allem die Erkennung des Fibroadenoms bedarf einer sorgfältigen Diagnosestellung, die des weiteren durch den Einsatz von Mayers- Hämalaun erleichtert werden soll. Die histologische Schnellschnittdiagnostik weist neben einer hohen Sensitivität eine optimale Spezifität auf und ist der zytologischen Methode im Erkennen von benignen Veränderungen überlegen. Da jedoch auch die histologische Schnellschnittdiagnose ihre Fehler hat, ist eine zusätzliche ergänzende zytologische Diagnosenstellung zu empfehlen. Bei Verwendung beider Methoden hätte bei allen 111 Patientinnen die richtige Diagnose gestellte werden können und ist somit aussagekräftigstes Argument für die parallele Verwendung dieser beiden Methoden.Cytologic intraoperative diagnosis of Breast Cancer Introduction The histological diagnosis of a suspect tumor of the breast is the gold standard in the intraoperative assessment. We know that each method has got its own limitations. So it is important to achieve the best treatment for the patient. We searched for a way to supplement the histologic clarification. Therefore we made an imprint cytology treated with a special quick stain identical to the intraoperative histologic examination in 111 female patients. These women are all patients of a University Womens Hospital with a specialized Breast Cancer Department. There are no other common than factors, other than the necessity of an operative clarification of a breast tumour. Preparation of the imprint During the surgery the pathologist gets the tissue which must be examined. The center of the tumor will be opened. By pressing softly on the center of the tumor, an imprint can be created. The main goal of this process is to have enough cells represented for a cytologic examination. To increase the possibility for a correct diagnose we prepared two imprints per patient. After this, the histologic examination can take place. Quick stain of imprint In relation to the established coloring methods like the MGG (May-Gruenwald-Giemsa) and the Papanicolaou stain, the quick stain needs only a few minutes to allow a cytologic diagnosis. This is the big advantage of this method. The imprint will be covered with a solution of MGG and methanol for two minutes. Then it will be rinsed with destillated water and then again covered for two minutes with a solution of Giemsa and destillated water. After clearing with destilled water the cytologic examination can be started. The cytologist has got no further information about the patient. When we compared the results of the histologic and the cytologic assessment, we saw that the cytologist had problems to judge fibroadenomas as benigne. So we improved the process by adding Mayers- Haemalaun as first step of the stain process to easier recognize this type of tissue. The Mayers- Haemalaun helps to identify the myoepithelial cells which is a sign for a non-malignant tissue. Results The diagnosis made by the cytologist and the diagnosis made during the surgery by the pathologist were compared with the final histologic diagnosis. The 111 examined tissues were in 93 cases malignant and in 18 cases benigne. The cytologic intraoperative diagnosis recognized 90 of 93 malignant and 14 non-malignant tumors. Two were false-positive, one was false negative and four samples could not be examined due to an insufficient amount of collected cells. The total sensitivity is 97% with a specifity of 78%. The histologic intraoperative diagnosis recognized 90 of 93 malignant and all 18 non-malignant tumors. The total sensitivity is 97% with a specifity of 100%. Conclusion The cytologic intraoperative diagnosis additionally to the histologic one is of benefit. Its high sensitivity depends on several factors. When making the imprint it is absolutely necessary to collect as many cells as possible. It is important for the cytological diagnosis to be accurate. Second, one has to realize the limitations of the cytologic diagnosis. The fibroadenoma for instance can be easily misinterpreted as a carcinoma. The Mayers- Haemalaun stain helps to differ between the fibroadenoma and a carcinoma. The histologic intraoperative diagnosis has got a high specifity as well as sensitivity. It recognizes non-malignant tumors easier than the cytologic one. However, each method has got its limitations. So it is recommendable to supplement the histologic assessment by a cytologic one. By using both methods, the correct diagnosis in all 111 patients could have been made. This is the best reason for using both methods together

Effective detection of BRAFV595E mutation in canine urothelial and prostate carcinomas using immunohistochemistry.

Canine urothelial carcinoma (UC) and prostate carcinoma (PC) frequently exhibit the BRAFV595E mutation, akin to the BRAFV600E mutation common in various human cancers. Since the initial discovery of the BRAF mutation in canine cancers in 2015, PCR has been the standard method for its detection in both liquid and tissue biopsies. Considering the similarity between the canine BRAFV595E and human BRAFV600E mutations, we hypothesized that immunohistochemistry (IHC) using a BRAFV600E-specific antibody could effectively identify the canine mutant BRAFV595E protein. We tested 122 canine UC (bladder n = 108, urethra n = 14), 21 PC, and benign tissue using IHC and performed digital droplet PCR (ddPCR) on all 122 UC and on 14 IHC positive PC cases. The results from ddPCR and IHC were concordant in 99% (135/136) of the tumours. Using IHC, BRAFV595E was detected in 72/122 (59%) UC and 14/21 (65%) PC. Staining of all benign bladder and prostate tissues was negative. If present, mutant BRAF staining was homogenous, with rare intratumour heterogeneity in three (4%) cases of UC. Additionally, the BRAFV595E mutation was more prevalent in tumours with urothelial morphology, and less common in glandular PC or UC with divergent differentiation. This study establishes that BRAFV600-specific IHC is a reliable and accurate method for detecting the mutant BRAFV595E protein in canine UC and PC. Moreover, the use of IHC, especially with tissue microarrays, provides a cost-efficient test for large-scale screening of canine cancers for the presence of BRAF mutations. This advancement paves the way for further research to define the prognostic and predictive role of this tumour marker in dogs and use IHC to stratify dogs for the treatment with BRAF inhibitors