Objective: Obesity induces hypothyroidism with unknown mechanisms. This study investigates the role of (Receptor for Hyaluronan-Mediated Motility (RHAMM) in obesity-associated thyroid dysfunction, focusing on hepatic oxidative stress. Methods: Global RHAMM-deficient mice and their wildtype littermate controls were fed a normal chow diet or high-fat diet (HFD) for 16 weeks. Thyroid function was evaluated by measuring plasma thyroid-stimulating hormone (TSH) levels. The hepatic oxidative response was assessed by measuring signaling pathways associated with nuclear factor erythroid 2-related factor 2 (Nrf2) activity. Results: HFD feeding increased plasma TSH levels in male mice but not in female mice. RHAMM deletion in male mice mitigated HFD-induced TSH elevation, which was associated with enhanced hepatic antioxidant defenses and reduced inflammation. This was evidenced by elevated expression of the Nrf2 target gene NAD(P)H: quinone oxidoreductase 1 (Nqo1), reduced protein carbonylation and nitration levels, and reduced expression of the pro-inflammatory cytokines IL-1β and TNF-α in livers of male RHAMM-deficient mice. Mechanistically, RHAMM deletion decreased AKT/ERK signaling, increased GSK3 signaling, increased CD44 protein expression, and increased Nqo1 levels in the liver. Conclusions: RHAMM promotes obesity-induced thyroid dysfunction by regulating oxidative stress and inflammation in male mice. Targeting RHAMM may provide a novel therapeutic strategy for mitigating obesity-related endocrine and metabolic disorders.

CD5 antigen-like (CD5L), also known as apoptosis inhibitor expressed by macrophages (AIM), is a secreted protein produced by macrophages, which has both cell-autonomous and non-autonomous functions: it has been implicated in multiple biological processes, including inflammation, infection, and fibrosis. The transcriptional regulation of CD5L is complex, and a comparison of the proteomes of bone marrow-derived macrophages from mice with graded expression of transcription factor NF-E2 p45-related factor 2 (Nrf2) indicated that Nrf2 controls expression of the CD5L gene. Using genetic and pharmacological means to increase or decrease Nrf2 activity, we found a correlation between the abundance of Nrf2 and CD5L expression in both murine macrophages and primary human monocyte-derived macrophages. Furthermore, the potency of small-molecule Nrf2 activators with distinct mechanisms of action and different potencies, correlated with the extent of CD5L expression, both at the mRNA and the secreted protein levels. Conversely, depletion of Nrf2 resulted in a significant decrease in CD5L mRNA levels. Chip-seq analysis showed Nrf2 binding 20,000 bp upstream of the Cd5l promoter, a region containing sequences resembling the antioxidant response element (ARE, 5'-TGACNNNGC-3') Nrf2-binding motif. Deletion of two of these sequences by CRISPR/Cas9 gene editing led to a profound decrease in CD5L mRNA levels, confirming the critical role of these ARE-like sequences in controlling CD5L expression. Recombinant CD5L (rCD5L) suppressed transforming growth factor β (TGFβ)-directed fibrogenic responses in human lung fibroblasts, suggesting that Nrf2 activators, such as the clinically used omaveloxolone (RTA-408), could protect against pulmonary fibrosis by boosting the levels of CD5L.

6-(Methylsulfinyl)hexyl isothiocyanate, here referred to as hexaraphane, is a bioactive compound found in wasabi, which exhibits cytoprotective, anti-inflammatory, and chemopreventive properties. The beneficial effects of hexaraphane are largely mediated through derepression of transcription factor Nrf2, which is typically repressed via proteasomal degradation mediated by its two-site interaction with the ubiquitin E3 ligase adapter Keap1. Like other isothiocyanates, hexaraphane increases Nrf2 activity by perturbing the Nrf2-Keap1 interaction and stalling Keap1-directed ubiquitination. However, we found that hexaraphane modestly increases Nrf2 levels in Keap1-deficient cells, suggesting an additional Keap1-independent mechanism. Unlike other electrophilic Nrf2 activators, hexaraphane did not significantly impact Erk, p38Mapk, Jnk, or Pten/Pi3k/Akt signaling. Instead, our data reveal that hexaraphane inhibits Gsk-3β, a kinase that targets Nrf2 for proteasomal degradation by phosphorylating a DSGIS degron in the transcription factor through which it interacts with the β-TrCP E3 ligase adaptor. Hexaraphane reduced Nrf2 ubiquitination and its binding to Gsk-3β, mimicking the effects of the Gsk-3β inhibitor SB216763. In vitro kinase assays confirmed that hexaraphane suppresses Nrf2 phosphorylation. Furthermore, simulations of molecular docking and dynamics predict a specific interaction between hexaraphane and the catalytic groove of Gsk-3β. These findings identify hexaraphane as a previously unrecognized dual-acting Nrf2 activator that stabilizes Nrf2 both by disturbing Keap1 binding and by inhibiting Gsk-3β that forms the DSGIpS338 phosphodegron.

Accumulation of reactive oxygen species (i.e., oxidative stress) is a leading cause of beta cell dysfunction and apoptosis in diabetes. NRF2 (NF-E2 p45-related factor-2) regulates the adaptation to oxidative stress, and its activity is negatively regulated by the redox-sensitive CUL3 (cullin-3) ubiquitin ligase substrate adaptor KEAP1 (Kelch-like ECH-associated protein-1). Additionally, NRF2 is repressed by the insulin-regulated Glycogen Synthase Kinase-3 (GSK3). We have demonstrated that phosphorylation of NRF2 by GSK3 enhances β-TrCP (beta-transducin repeat-containing protein) binding and ubiquitylation by CUL1 (cullin-1), resulting in increased proteasomal degradation of NRF2. Thus, we hypothesise that inhibition of GSK3 activity or β-TrCP binding upregulates NRF2 and so protects beta cells against oxidative stress. We have found that treating the pancreatic beta cell line INS-1 832/13 with the KEAP1 inhibitor TBE31 significantly enhanced NRF2 protein levels. The presence of the GSK3 inhibitor CT99021 or the β-TrCP-NRF2 protein-protein interaction inhibitor PHAR, along with TBE31, resulted in prolonged NRF2 stability and enhanced nuclear localisation (P < 0.05). TBE31-mediated induction of NRF2-target genes encoding NAD(P)H quinone oxidoreductase 1 (Nqo1), glutamate-cysteine ligase modifier (Gclm) subunit and heme oxygenase (Hmox1) was significantly enhanced by the presence of CT99021 or PHAR (P < 0.05) in both INS-1 832/13 and in isolated mouse islets. Identical results were obtained using structurally distinct GSK3 inhibitors and inhibition of KEAP1 with sulforaphane. In summary, we demonstrate that GSK3 and β-TrCP/CUL1 regulate the proteasomal degradation of NRF2, enhancing the impact of KEAP1 regulation, and so contributes to the redox status of pancreatic beta cells. Inhibition of GSK3, or β-TrCP/CUL1 binding to NRF2 may represent a strategy to protect beta cells from oxidative stress.

At presentation, approximately 25% of patients will have muscle-invasive bladder cancer (MIBC), and patients with non-MIBC may subsequently progress to MIBC. Prognosis is dependent on TNM stage (Table 12.1) and pelvic lymph node status. The disease requires a multi-disciplinary approach, encompassing surgeon, oncologist, radiologist, pathologist and cancer specialist nurse.

PDF file - 572K, Cellular co-localization of ectopic GFP and RFP in HEK293T cells.

Abstract Introduction Blue light cystoscopy with hexaminolevulinate (HAL) during transurethral resection of bladder cancer (TURBT) has been shown to improve detection, thereby reducing bladder cancer recurrence compared with white light cystoscopy. Methods Single‐centred UK (United Kingdom) study on 101 patients who underwent blue light cystoscopy between July 2017 and November 2020, performed by a single surgeon. Our study was divided into two arms; the primary arm had no prior diagnosis of bladder malignancy (N = 41), whereas secondary re‐resection arm had (N = 57). Three patients with non‐urothelial bladder cancer were excluded. Patients were followed up for 24 months. Data were collected on biopsy quality, histopathological characteristics and recurrence. The end points of the study were recurrence rate at 24 months in both arms and detection of CIS in patients who undergo TURBT or biopsy after initial white light study in the secondary, re‐resection arm. This was analysed with Fisher's exact test. Results Of 98 patients, 39 had malignancy in their first blue light TURBT/biopsy: primary arm (10/41, 24.4%) and secondary arm (29/57, 50.9%), with detrusor present in 80.5% and 80.7%, respectively. In the secondary arm, blue light re‐resection TURBT detected significantly more CIS (20.7% vs 51.7%, p = 0.0277) compared with white light with 3.4% upstaged to muscle invasive bladder cancer (G3pT2). Median time to re‐resection was 3.06 months. Recurrence rate was 33.3% in the primary arm and 37.5% in the secondary arm after 24 months of follow‐up. Conclusion Our data confirm that blue light TURBT with HAL provides superior detection and diagnosis of CIS in patients with previous white light cystoscopy.

Supplementary Figure 1 from Generation of a Stable Antioxidant Response Element–Driven Reporter Gene Cell Line and Its Use to Show Redox-Dependent Activation of Nrf2 by Cancer Chemotherapeutic Agents

PDF file - 288K, The binding of NRF2 to the ARE sites in MCF7 and MCF7-RXRo cells.

PDF file - 232K, BHA up-regulated ARE-gene battery in mouse small intestine and liver.

PDF file - 160K, tBHQ increased the expression of AKR1C in MCF7 cells.

PDF file - 499K, Recombinant his-RXRa was unable to interact with GST-hNRF2 mutant fusion protein that lacked residues 209-316 between the Neh5 and Neh6 domains.

PDF file - 97K, Includes plasmid construction data, GST pull-down assay, and immunoprecipitation data. Also includes supplementary references.

The move towards transperineal biopsy and retreat from the traditional transrectal approach termed ‘TREXIT’ is in vogue. The reduced sepsis and emergency readmission rates associated with the transperineal approach were alluded to in our paper reviewing the clinical and financial implications of a decade of NHS prostate biopsies from 2008 to 2019 [1]. The recent 2022 European Association of Urology Guidelines on Prostate Cancer now strongly recommend the use of the transperineal approach due to the lower risk of infectious complications [2]. During the last 2 years of our original cohort (2017–2019) the majority of biopsies were transrectal (67.2%). The COVID-19 pandemic has had a profound impact on the delivery of cancer services. The aim of this updated review was to evaluate the most recent NHS prostate biopsy data from 2019 to 2021 in the context of a global pandemic.

PDF file - 188K, RXRa interacted with Nrf2 in vivD.

PDF file - 239K, The interaction of mNrf2 with RXRa in MCF7-RXRa cells.

A 56yearold otherwise healthy female presented in 2020 with right axillary lymphadenopathy. She had had bilateral breast augmentation in early 2000 with placement of bilateral textured implants. A positron emission tomography (18FFDG PET) scan showed a 2.4 cm hypermetabolic right axillary lymph node. Biopsy demonstrated anaplastic large cell lymphoma (ALCL), ALKnegative, T/null cell type (upper left, all microscopic images, objective × 40). Subsequent right breast biopsy demonstrated that these large neoplastic cells were positive for CD30 (upper right), granzyme B, TIA1 and p63. Clonal Tcell receptor gene rearrangement was detected, without TP63 or DUSP22 rearrangement. Bone marrow biopsy showed that lymphoma cells constituted approximately 1% of bone marrow cells (lower left). From May 2020 to September 2020, the patient underwent six 3weekly cycles of immunochemotherapy with cyclophosphamide, doxorubicin, prednisone and brentuximab vedotin. Posttreatment 18FFDG PET scan showed a complete response. Subsequently, the patient underwent bilateral capsulectomies. Intraoperatively, a “velvety” appearance of exudative, fibrinous tissue was noted on the surface of the right capsule, surrounded by brown, turbid fluid (lower right). The patient has been followed with serial PET scans and remains in remission. Breast implantassociated ALCL (BIAALCL) is a rare type of Tcell nonHodgkin lymphoma. The prognosis is favourable in the majority of patients, who have localised disease. Bone marrow involvement is uncommon. The addition of brentuximab vedotin, an antiCD30 antibody conjugate, to cyclophosphamide, doxorubicin and prednisone (ACHP) for treatment of CD30positive BIAALCL should be considered as a systemic regimen in combination with local therapy by capsulectomies.

The adoption of minimally invasive laparoscopic techniques has revolutionised urological practice. This necessitates a pneumoperitoneum (PNP) and the impact the PNP pressure has on post-operative outcomes is uncertain. During the current COVID-19 era guidance has suggested the utilisation of lower PNP pressures to mitigate the risk of intra-operative viral transmission. Review the current literature regarding the impact of pneumoperitoneum pressure, within the field of urology, on post-operative outcomes. A search of the PubMed, Medline and EMBASE databases was undertaken to identify studies that met the inclusion criteria. The Preferred Reporting Items for Systematic Reviews (PRISMA) guidelines were adhered to. Ten studies, that included both randomised controlled trials and retrospective case series reviews, were identified that met the inclusion criteria. The effect of PNP pressure on outcomes following prostatectomy, live donor nephrectomy, partial nephrectomy and a variety of benign upper tract procedures were discussed. Low pressure PNP appears safe when compared to high pressure PNP, potentially reducing post-operative pain and rates of ileus. When compared to general surgery, there is a lack of quality evidence investigating the impact of PNP pressures on outcomes within urology. Low pressure PNP appears non-inferior to high pressure PNP. More research is required to validate this finding, particularly post-cystectomy and nephrectomy.