Activation of NOTCH signaling impedes cell proliferation and survival in acute megakaryoblastic leukemia.

The genetic lesions that drive acute megakaryoblastic leukemia (AMKL) have not been fully elucidated. To search for genetic alterations in AMKL, we performed targeted deep sequencing in 34 AMKL patient samples and 8 AMKL cell lines and detected frequent genetic mutations in the NOTCH pathway in addition to previously reported alterations in GATA-1 and the JAK-STAT pathway. Pharmacological and genetic NOTCH activation, but not inhibition, significantly suppressed AMKL cell proliferation in both in vitro and in vivo assays employing a patient-derived xenograft model. These results suggest that NOTCH inactivation underlies AMKL leukemogenesis. and NOTCH activation holds the potential for therapeutic application in AMKL.

The design and validation of a low-cost trans perineal (TP) prostate biopsy simulator for training: improving trainees' confidence and cognitive targeting skills.

PURPOSE: The aim of this research was to create a novel and low-cost TP prostate biopsy simulator that has face, content and construct validity with high educational value. METHODS: This research developed a trans perineal prostate (TP) biopsy simulator using 3D-printed moulds and tissue-mimicking materials. Important regions (anterior, mid, and posterior zones) were coded with different colours. Ultrasound visible abnormal lesions were embedded in the prostate phantom. Expert and novice participants in TP biopsies were recruited. Essential skills were identified through the consensus of six experts. These skills were assessed through tasks performed by participants. This included the accuracy and timing of systematic and target biopsies. Immediate feedback was determined by the colour of the biopsy cores taken. A survey was distributed to evaluate its realism and educational value. RESULTS: The material cost of one simulator was £7.50. This simulator was proven to have face, content, and construct validity. There was a significant difference (p = 0.02) in the accuracy of systematic biopsies between both experts and novices. Significant difference was also observed (p = 0.01), in accurately identifying target lesion on ultrasound between both groups. Participants rated the overall realism of the simulator 4.57/5 (range 3-5). 100% of the experts agreed that introducing this simulator to training will be beneficial. 85.7% of the participants strongly agree that the simulator improved their confidence in TP biopsies. CONCLUSION: There is value in integrating this proof-of-concept TP prostate biopsy simulator into training. It has highly rated educational value and has face, content, and construct validity.

Transfer of open and laparoscopic skills to robotic surgery: a systematic review.

Due to its advantages over open surgery and conventional laparoscopy, uptake of robot-assisted surgery has rapidly increased. It is important to know whether the existing open or laparoscopic skills of robotic novices shorten the robotic surgery learning curve, potentially reducing the amount of training required. This systematic review aims to assess psychomotor skill transfer to the robot in clinical and simulated settings. PubMed, EMBASE, Cochrane Library and Scopus databases were systematically searched in accordance with PRISMA guidelines from inception to August 2021 alongside website searching and citation chaining. Article screening, data extraction and quality assessment were undertaken by two independent reviewers. Outcomes included simulator performance metrics or in the case of clinical studies, peri- and post-operative metrics. Twenty-nine studies met the eligibility criteria. All studies were judged to be at high or moderate overall risk of bias. Results were narratively synthesised due to heterogeneity in study designs and outcome measures. Two of the three studies assessing open surgical skill transfer found evidence of successful skill transfer while nine of twenty-seven studies evaluating laparoscopic skill transfer found no evidence. Skill transfer from both modalities is most apparent when advanced robotic tasks are performed in the initial phase of the learning curve but quality and methodological limitations of the existing literature prevent definitive conclusions. The impact of incorporating laparoscopic simulation into robotic training curricula and on the cost effectiveness of training should be investigated.

Radiation Safety Knowledge and Practice in Urology Theaters: A Collaborative Multicenter Survey.

Objective: To evaluate the knowledge and current radiation safety practice among health care professionals undertaking fluoroscopic procedures in urology. Materials and Methods: A 14-question survey was disseminated to multidisciplinary urology theater staff. Questions included demographic data, usual radiation safety practice, and knowledge. The questions were selected based on guidelines from the International Commission of Radiological Protection and Health and Safety Executive. The survey was disseminated through regional collaborators and social media. Results: The survey received a total of 309 completed responses, including 272 from the United Kingdom. Responses from the United Kingdom multidisciplinary team included 164 (60.3%) urologic surgeons, 68 (25.0%) theater nurses, 27 (9.9%) from the anesthetic team, and 13 (4.7%) radiographers. Results from the United Kingdom demonstrated use of lead aprons and thyroid shields as 99.3% and 52.2%, respectively. Lead glasses and lead glove use were 7.4% and 0.7%, respectively. Lack of availability was cited as a reason for noncompliance with shielding guidelines in 208 (76.5%) of the respondents. No form of training in radiation safety was reported by 120 (44.1%) respondents. However, there was no association between answering knowledge questions correctly and having completed some form of radiation safety training (p = 0.41). There was an association between dosimeter use and those who had received radiation safety training (p = 0.02). Consultant urologists were also more likely to use a dosimeter than training grade urologists (p = 0.035). Conclusion: Suboptimal compliance with radiation safety guidelines is prevalent in contemporary urologic practice, and presents a significant occupational health concern. Availability of protective equipment needs to be urgently addressed.

Triage of urology service to cope with COVID-19 pandemic: A single institution study.

Almost a year ago, no one has ever heard of COVID-19 but now, every individual in the world is familiar with this term. It is far from over and yet, it has affected every aspect of human life. The Department of Urology at King's College Hospital London provides all types of urology care ranging from benign to cancer treatments to the community. However, this service was badly affected by COVID-19. Policies were made by the experts in the field to reduce patient traffic in the hospital and at the same time, attempting to ensure appropriate and timely treatment was provided to patients suffering from urological conditions requiring urgent attention. In this article, we discuss the triage guidelines set up at our centre. Treatments for benign conditions such as kidney stones were delayed for 3-6 months. For the first time, telephone and video clinics were setup to follow-up patients with benign conditions. Urological emergencies such as acute urinary retention and priapism were discharged from accidental and emergency department after treatment. Small T1 renal cancers were put on surveillance, whereas T2 and T3 renal cancers were offered nephrectomy at a COVID-free specialized center. Transurethral removal of bladder tumor was offered only for solid or actively bleeding tumor. High risk prostate cancer patients were started on hormonal therapy and radiotherapy was only offered for spinal cord compression secondary to metastasis. Low and intermediate non-metastatic prostate cancers were placed on active surveillance. Patients with testicular tumor continued to have immediate inguinal orchidectomy. The multi-disciplinary meetings were done remotely using blue jeans software®. These steps not only strive to provide adequate and timely urology care to patients but also protect health care workers and prevent the spread of COVID-19.

The pro-metastasis tyrosine phosphatase, PRL-3 (PTP4A3), is a novel mediator of oncogenic function of BCR-ABL in human chronic myeloid leukemia.

BACKGROUND: Resistance to tyrosine kinase inhibitors (TKIs) remains a challenge in management of patients with chronic myeloid leukemia (CML). A better understanding of the BCR-ABL signalling network may lead to better therapy. FINDINGS: Here we report the discovery of a novel downstream target of BCR-ABL signalling, PRL-3 (PTP4A3), an oncogenic tyrosine phosphatase. Analysis of CML cancer cell lines and CML patient samples reveals the upregulation of PRL-3. Inhibition of BCR-ABL signalling either by Imatinib or by RNAi silencing BCR-ABL reduces PRL-3 and increases cleavage of PARP. In contrast, the amount of PRL-3 protein remains constant or even increased in response to Imatinib treatment in drug resistant cells expressing P210 T315I. Finally, analysis with specific shRNA shows PRL-3 involvement in the proliferation and self-renewal of CML cells. CONCLUSIONS: These data support a role for PRL-3 in BCR-ABL signalling and CML biology and may be a potential therapeutic target downstream of BCR-ABL in TKI resistant mutant cells.

Dual inactivation of Hus1 and p53 in the mouse mammary gland results in accumulation of damaged cells and impaired tissue regeneration.

In response to DNA damage, checkpoint proteins halt cell cycle progression and promote repair or apoptosis, thereby preventing mutation accumulation and suppressing tumor development. The DNA damage checkpoint protein Hus1 associates with Rad9 and Rad1 to form the 9-1-1 complex, which localizes to DNA lesions and promotes DNA damage signaling and repair. Because complete inactivation of mouse Hus1 results in embryonic lethality, we developed a system for regulated Hus1 inactivation in the mammary gland to examine roles for Hus1 in tissue homeostasis and tumor suppression. Hus1 inactivation in the mammary epithelium resulted in genome damage that induced apoptosis and led to depletion of Hus1-null cells from the mammary gland. Conditional Hus1 knockout females retained grossly normal mammary gland morphology, suggesting compensation by cells that failed to undergo Cre-mediated Hus1 deletion. p53-deficiency delayed the clearance of Hus1-null cells from conditional Hus1 knockout mice and caused the accumulation of damaged, dying cells in the mammary gland. Notably, compensatory responses were impaired following combined Hus1 and p53 loss, resulting in aberrant mammary gland morphology and lactation defects. Overall, these results establish a requirement for Hus1 in the survival and proliferation of mammary epithelium and identify a role for p53 in mammary gland tissue regeneration and homeostasis.

Humanization of a chicken anti-IL-12 monoclonal antibody.

Chicken anti-IL-12 monoclonal antibodies were isolated by phage display using spleen cells from a chicken immunized with human and mouse IL-12 as a source for library construction. One of the chicken monoclonal antibodies, DD2, exhibited binding to both human and mouse IL-12 in the single-chain Fv form and also after conversion to chicken-human chimeric IgG1/lambda antibody. The chicken DD2 variable regions were humanized by transferring their CDRs and several framework amino acids onto human acceptor variable regions. In the Vlambda, six chicken framework amino acids were identified to be important for the conformation of the CDR structure by computer modeling and therefore were retained in the humanized form; likewise, five chicken amino acids in the VH framework regions were retained in the humanized VH. The affinities of humanized DD2 IgG1/lambda to human and mouse IL-12 measured by competitive binding were nearly identical to those of chicken-human chimeric DD2 IgG1/lambda. This work demonstrates that humanization of chicken monoclonal antibodies assisted by computer modeling is possible, leading to a new way to generate therapeutic humanized antibodies against antigens to which the rodent immune system may fail to efficiently raise high affinity antibodies.

Engineered human IgG antibodies with longer serum half-lives in primates.

The neonatal Fc receptor (FcRn) plays an important role in regulating the serum half-lives of IgG antibodies. A correlation has been established between the pH-dependent binding affinity of IgG antibodies to FcRn and their serum half-lives in mice. In this study, molecular modeling was used to identify Fc positions near the FcRn binding site in a human IgG antibody that, when mutated, might alter the binding affinity of IgG to FcRn. Following mutagenesis, several IgG2 mutants with increased binding affinity to human FcRn at pH 6.0 were identified at Fc positions 250 and 428. These mutants do not bind to human FcRn at pH 7.5. A pharmacokinetics study of two mutant IgG2 antibodies with increased FcRn binding affinity indicated that they had serum half-lives in rhesus monkeys approximately 2-fold longer than the wild-type antibody.