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Platinum-based chemotherapy has been the standard first-line (1L) treatment for advanced urothelial carcinoma (UC) for decades, based on the proven efficacy and established safety profiles of cisplatin- and carboplatin-based regimens. With the emergence of novel regimens, it is important to reevaluate and contextualize the role of 1L platinum-based chemotherapy. Platinum-based chemotherapy followed by avelumab 1L maintenance in patients without disease progression following platinum-based chemotherapy was established as a standard 1L regimen based on the JAVELIN Bladder 100 phase III trial. More recently, the EV-302 phase III trial showed the superiority of 1L enfortumab vedotin (EV) + pembrolizumab versus platinum-based chemotherapy, and the Checkmate 901 phase III trial showed the superiority of 1L nivolumab + cisplatin/gemcitabine versus cisplatin/gemcitabine alone. These 2 regimens have now been included as standard 1L options in treatment guidelines for advanced UC. EV + pembrolizumab is now the preferred 1L treatment, and in locations where EV + pembrolizumab is not available or individual patients are not considered suitable, recommended options are platinum-based chemotherapy followed by avelumab maintenance or nivolumab + cisplatin-based chemotherapy. In this review, we discuss current treatment options for advanced UC recommended in guidelines, practical considerations with platinum-based chemotherapy, the role of avelumab 1L maintenance, recent phase III trials of EV + pembrolizumab and nivolumab + cisplatin/gemcitabine, safety profiles of recommended 1L treatments, and second-line treatment options., (© The Author(s) 2024. Published by Oxford University Press.)

Background and Objective: Molecular classification of upper tract urothelial carcinoma (UTUC) can provide insight into divergent clinical outcomes and provide a biological rationale for clinical decision-making. As such, we performed multi-omic analysis of UTUC tumors to identify molecular features associated with disease recurrence and response to immune checkpoint blockade (ICB)., Methods: Targeted DNA and whole transcriptome RNA sequencing was performed on 100 UTUC tumors collected from patients undergoing nephroureterectomy. Consensus non-negative matrix factorization was used to identify molecular clusters associated with clinical outcomes. Gene set enrichment and immune deconvolution analyses were performed. Weighted gene co-expression network analysis was employed for unsupervised identification of gene networks in each cluster., Key Findings and Limitations: Five molecular clusters with distinct clinical outcomes were identified. Favorable subtypes (C1 and C2) were characterized by a luminal-like signature and an immunologically depleted tumor microenvironment (TME). Subtype C3 was characterized by FGFR3 alterations and a higher tumor mutational burden, and included all tumors with microsatellite instability. Despite higher rates of recurrence and inferior survival, subtypes C4 and C5 harbored an immunologically rich TME favoring response to ICB. Limitations include extrapolation of molecular features of tumors from the primary site to determine response to systemic immunotherapy and the limited resolution of bulk sequencing to distinguish gene expression in the tumor, stroma, and immune compartments., Conclusions and Clinical Implications: RNA sequencing identified previously underappreciated UTUC molecular heterogeneity and suggests that UTUC patients at the highest risk of metastatic recurrence following surgery include those most likely to benefit from perioperative ICB., (Copyright © 2024. Published by Elsevier B.V.)

Background: There is no doubt that global warming, with its extreme heat events, is having an increasing impact on human health. Heat is not independent of ambient temperature but acts synergistically with relative humidity (RH) to increase the risk of several diseases, such as cardiovascular and pulmonary diseases. Although the skin is the organ in direct contact with the environment, it is currently unknown whether skin health is similarly affected. Objective: While mechanistic studies have demonstrated the mechanism of thermal aging, this is the first epidemiological study to investigate the effect of long-term exposure to heat index (HI) as a combined function of elevated ambient temperature and RH on skin aging phenotypes in Indian women. Methods: The skin aging phenotypes of 1510 Indian women were assessed using the Score of Intrinsic and Extrinsic Skin Aging (SCINEXA™) scoring tool. We used data on ambient temperature and RH, combined into an HI with solar ultraviolet radiation (UVR), and air pollution (particulate matter <2.5 µm [PM 2.5] ; nitrogen dioxide [NO 2 ]) from secondary data sources with a 5-year mean residential exposure window. An adjusted ordinal multivariate logistic regression model was used to assess the effects of HI on skin aging phenotypes. Results: HI increased pigmentation such as hyperpigmented macula on the forehead (odds ratios [OR]: 1.31, 95% confidence interval [CI]: 1.12, 1.54) and coarse wrinkles such as crow's feet (OR: 1.17, 95% CI: 1.05, 1.30) and under-eye wrinkles (OR: 1.3, 95% CI: 1.15, 1.47). These associations were robust to the confounding effects of solar UVR and age. Prolonged exposure to extreme heat, as indicated by high HI, contributes to skin aging phenotypes. Conclusion: Thus, ambient temperature and RH are important factors in assessing the skin aging exposome.

Purpose: Alterations in DNA damage response (DDR) genes, including ERCC2 , have been correlated with response to neoadjuvant cisplatin-based chemotherapy (NAC) in patients with muscle-invasive bladder cancer (MIBC). The SWOG 1314 (S1314) trial enrolled patients with MIBC who received one of two NAC regimens followed by radical cystectomy. We examined the prevalence of DDR alterations in NAC responders versus nonresponders and correlated DDR alteration status with response., Methods: Pretreatment tumor specimens from 179 evaluable patients underwent next-generation sequencing (Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets assay). Associations were determined between any or only deleterious alterations within nine predefined DDR genes, or any alterations in ERCC2 , and progression-free survival (PFS) and overall survival using Cox regression, and, in a subset of evaluable patients, pathologic response (complete response, pT0, or downstaging to

Small cell carcinomas (SMC) of the lung are now molecularly classified based on the expression of transcriptional regulators (NEUROD1, ASCL1, POU2F3, and YAP1) and DLL3, which has emerged as an investigational therapeutic target. PLCG2 has been shown to identify a distinct subpopulation of lung SMC with stem cell-like and prometastasis features and poor prognosis. We analyzed the expression of these novel neuroendocrine markers and their association with traditional neuroendocrine markers and patient outcomes in a cohort of bladder neuroendocrine carcinoma (NEC) consisting of 103 SMC and 19 large cell NEC (LCNEC) assembled in tissue microarrays. Coexpression patterns were assessed and integrated with detailed clinical annotation including overall (OS) and recurrence-free survival (RFS) and response to neoadjuvant/adjuvant chemotherapy. We identified 5 distinct molecular subtypes in bladder SMC based on the expression of ASCL1, NEUROD1, and POU2F3: ASCL1+/NEUROD1- (n = 33; 34%), ASCL1- /NEUROD1+ (n = 21; 21%), ASCL1+/NEUROD1+ (n = 17; 17%), POU2F3+ (n = 22, 22%), and ASCL1- /NEUROD1- /POU2F3- (n = 5, 5%). POU2F3+ tumors were mutually exclusive with those expressing ASCL1 and NEUROD1 and exhibited lower expression of traditional neuroendocrine markers. PLCG2 expression was noted in 33 tumors (32%) and was highly correlated with POU2F3 expression (P < .001). DLL3 expression was high in both SMC (n = 72, 82%) and LCNEC (n = 11, 85%). YAP1 expression was enriched in nonneuroendocrine components and negatively correlated with all neuroendocrine markers. In patients without metastatic disease who underwent radical cystectomy, PLCG2+ or POU2F3+ tumors had shorter RFS and OS (P < .05), but their expression was not associated with metastasis status or response to neoadjuvant/adjuvant chemotherapy. In conclusion, the NEC of the bladder can be divided into distinct molecular subtypes based on the expression of ASCL1, NEUROD1, and POU2F3. POU2F3-expressing tumors represent an ASCL1/NEUROD1-negative subset of bladder NEC characterized by lower expression of traditional neuroendocrine markers. Marker expression patterns were similar in SMC and LCNEC. Expression of PLCG2 and POU2F3 was associated with shorter RFS and OS. DLL3 was expressed at high levels in both SMC and LCNEC of the bladder, nominating it as a potential therapeutic target., (Copyright © 2024 United States & Canadian Academy of Pathology. Published by Elsevier Inc. All rights reserved.)

Competing Interests: There are no conflicts of interest.

Background and Objective: Checkpoint inhibitor therapy (CPI) has demonstrated survival benefits in urothelial carcinoma (UC); however, not all patients benefit from CPI due to resistance. Combining sitravatinib, a multitargeted receptor tyrosine kinase inhibitor of TYRO3, AXL, and MERTK (TAM) receptors and VEGFR2, with CPI may improve antitumor responses. Our objective was to assess the efficacy and safety of sitravatinib plus nivolumab in patients with advanced/metastatic UC., Methods: The 516-003 trial (NCT03606174) is an open-label, multicohort phase 2 study evaluating sitravatinib plus nivolumab in patients with advanced/metastatic UC enrolled in eight cohorts depending on prior treatment with CPI, platinum-based chemotherapy (PBC), or antibody-drug conjugate (ADC). Overall, 244 patients were enrolled and treated with sitravatinib plus nivolumab (median follow-up 14.1-38.2 mo). Sitravatinib (free-base capsules 120 mg once daily [QD] or malate capsule 100 mg QD) plus nivolumab (240 mg every 2 wk/480 mg every 4 wk intravenously)., Key Findings and Limitations: The primary endpoint was objective response rate (ORR; RECIST v1.1). The secondary endpoints included progression-free survival (PFS) and safety. The Predictive probability design and confidence interval methods were used. Among patients previously treated with PBC, ORR, and median PFS were 32.1% and 3.9 mo in CPI-naïve patients (n = 53), 14.9% and 3.9 mo in CPI-refractory patients (n = 67), and 5.4% and 3.7 mo in CPI- and ADC-refractory patients (n = 56), respectively. Across all cohorts, grade 3 treatment-related adverse events (TRAEs) occurred in 51.2% patients and grade 4 in 3.3%, with one treatment-related death (cardiac failure). Immune-related adverse events occurred in 50.4% patients. TRAEs led to sitravatinib/nivolumab discontinuation in 6.1% patients., Conclusions and Clinical Implications: Sitravatinib plus nivolumab demonstrated a manageable safety profile but did not result in clinically meaningful ORRs in patients with advanced/metastatic UC in the eight cohorts studied., Patient Summary: In this study, the combination of two anticancer drugs, sitravatinib and nivolumab, resulted in manageable side effects but no meaningful responses in patients with bladder cancer., (Copyright © 2023 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

The incineration fly ash (IFA) resulting from municipal solid waste combustion is laden with heavy metals, necessitating proper treatment not only for environmental management but also to reclaim the metal values. The surge in non-traditional metals like cobalt as emerging contaminant within IFA samples further attracts to address this issue. In response, the hydrometallurgical recycling of a cobalt-bearing IFA has been studied. Thereby, approximately 98 % zinc and 96 % cobalt were leached using a 1.0 mol/L H 2 SO 4 solution at 90 °C and 1 h of leaching time. In-depth analysis of the leaching process unveiled metals' dissolution primarily via the ion-exclusion mechanism, as evidenced by lower diffusion coefficients (between 10 -9 and 10 -11  m 2 /s) and activation energies (9.6-14.9 kJ/mol). Above 99 % separation of zinc from the cobalt-bearing leach liquor was achieved by extraction with 1.0 mol/L D2EHPA at an equilibrium pH below 3.0, followed by stripping with a 2.0 mol/L H 2 SO 4 solution. Cobalt, remained in the raffinate was efficiently precipitated by adding a 20 % excess dosage of oxalic acid to the stoichiometric ratio of C 2 O 4 2- :Co 2+ , resulting in only 5 mg/L cobalt left in the solution when precipitation occurred at a pH of 2.8. Additionally, the conversion of CoC 2 O 4 to high-purity Co 3 O 4 was conducted through heat-treatment at 600 °C. The resulting Co 3 O 4 was mixed with Li 2 CO 3 precursor that exhibited good electrochemical properties with a capacity of 128 mAh/g, thus affirming the high quality of the recycled cobalt. A comprehensive life-cycle assessment of the recycling process revealed that cobalt precipitation alone contributes approximately 50 % of the total global warming potential (GWP = 4.2624 kg CO 2 precursor that exhibited good electrochemical properties with a capacity of 128 mAh/g, thus affirming the high quality of the recycled cobalt. A comprehensive life-cycle assessment of the recycling process revealed that cobalt precipitation alone contributes approximately 50 % of the total global warming potential (GWP = 4.2624 kg CO 2 -eq). Notably, this value is remarkably lower than the GWP reported for primary cobalt production, highlighting the environmentally-friendly approach of this recycling endeavor., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Emerging organic contaminants present in the environment can be biodegraded in anodic biofilms of microbial fuel cells (MFCs). However, there is a notable gap existing in deducing the degradation mechanism, intermediate products, and the microbial communities involved in degradation of broad-spectrum antibiotic such as triclosan (TCS). Herein, the possible degradation of TCS is explored using TCS acclimatized biofilms in MFCs. 95% of 5 mgL -1 TCS are been biodegraded within 84 h with a chemical oxygen demand (COD) reduction of 62% in an acclimatized-MFC (A-MFC). The degradation of TCS resulted in 8 intermediate products including 2,4 -dichlorophenol which gets further mineralized within the system. Concurrently, the 16S rRNA V3-V4 sequencing revealed that there is a large shift in microbial communities after TCS acclimatization and MFC operation. Moreover, 30 dominant bacterial species (relative intensity >1%) are identified in the biofilm in which Sulfuricurvum kujiense, Halomonas phosphatis, Proteiniphilum acetatigens, and Azoarcus indigens significantly contribute to dihydroxylation, ring cleavage and dechlorination of TCS. Additionally, the MFC was able to produce 818 ± 20 mV voltage output with a maximum power density of 766.44 mWm -2 . The antibacterial activity tests revealed that the biotoxicity of TCS drastically reduced in the MFC effluent, signifying the non-toxic nature of the degraded products. Hence, this work provides a proof-of-concept strategy for sustainable mitigation of TCS in wastewaters with enhanced bioelectricity generation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Photophysics of a blue light-emitting fluorescent random copolymer, consisting of arylated polydioctylfluorene (aryl-F8), polydioctylfluorene (F8), and amine comonomers in a ratio of 80:15:5 is reported. In a solution of 10 -6 M, solvatochromism in absorption and photoluminescence (PL) is observed with an increased lifetime of PL as the polarity of the solvent increases. Dual fluorescence is observed in the 10 -9 M diluted solution, comprising a structured emission from a localized state in the aryl-F8 comonomer and a broad emission peak from the charge-transfer (CT) state at a lower energy. Emission wavelength-dependent time-resolved photoluminescence studies in different polar media confirm the presence of the emissive intrachain CT state in this copolymer. Analyzing the PL decay kinetics, we calculated the formation rate of the intrachain CT state to be ∼3.0 × 10 9 s -1 . Repopulation of the localized state from the CT state is observed in the lower polarity medium with a rate of 7 × 10 8 s -1 , which is almost absent for the large Stokes-shifted CT emission in the higher polarity medium. Along with the fundamental understanding of the photophysics of the random copolymer, this study suggests that the emission spectrum can be tailored by the concentration of polymer and the polarity of surrounding media.

Purpose: Patients with residual invasive bladder cancer after neoadjuvant chemotherapy (NAC) and radical cystectomy have a poor prognosis. Data on adjuvant therapy for these patients are conflicting. We sought to evaluate the natural history and genomic landscape of chemotherapy-resistant bladder cancer to inform patient management and clinical trials., Methods: Data were collected on patients with clinically localized muscle-invasive urothelial bladder cancer treated with NAC and cystectomy at our institution between May 15, 2001, and August 15, 2019, and completed four cycles of gemcitabine and cisplatin NAC, excluding those treated with adjuvant therapies. Survival was estimated using the Kaplan-Meier method, and multivariable Cox proportional hazards models were used to identify predictors of recurrence-free survival (RFS). Genomic alterations were identified in targeted exome sequencing (Memorial Sloan Kettering Integrated Mutation Profiling of Actionable Cancer Targets) data from post-NAC specimens from a subset of patients., Results: Lymphovascular invasion (LVI) was the strongest predictor of RFS (hazard ratio, 2.15 [95% CI, 1.37 to 3.39]) on multivariable analysis. Patients with ypT2N0 disease without LVI had a significantly prolonged RFS compared with those with LVI (70% RFS at 5 years). Lymph node yield did not affect RFS. Among patients with sequencing data (n = 101), chemotherapy-resistant tumors had fewer alterations in DNA damage response genes compared with tumors from a publicly available chemotherapy-naïve cohort (15% v 29%; P = .021). Alterations in CDKN2A/B were associated with shorter RFS. PIK3CA alterations were associated with LVI. Potentially actionable alterations were identified in more than 75% of tumors., Conclusion: Although chemotherapy-resistant bladder cancer generally portends a poor prognosis, patients with organ-confined disease without LVI may be candidates for close observation without adjuvant therapy. The genomic landscape of chemotherapy-resistant tumors is similar to chemotherapy-naïve tumors. Therapeutic opportunities exist for targeted therapies as adjuvant treatment in chemotherapy-resistant disease.

Background: No treatment has surpassed platinum-based chemotherapy in improving overall survival in patients with previously untreated locally advanced or metastatic urothelial carcinoma., Methods: We conducted a phase 3, global, open-label, randomized trial to compare the efficacy and safety of enfortumab vedotin and pembrolizumab with the efficacy and safety of platinum-based chemotherapy in patients with previously untreated locally advanced or metastatic urothelial carcinoma. Patients were randomly assigned in a 1:1 ratio to receive 3-week cycles of enfortumab vedotin (at a dose of 1.25 mg per kilogram of body weight intravenously on days 1 and 8) and pembrolizumab (at a dose of 200 mg intravenously on day 1) (enfortumab vedotin-pembrolizumab group) or gemcitabine and either cisplatin or carboplatin (determined on the basis of eligibility to receive cisplatin) (chemotherapy group). The primary end points were progression-free survival as assessed by blinded independent central review and overall survival., Results: A total of 886 patients underwent randomization: 442 to the enfortumab vedotin-pembrolizumab group and 444 to the chemotherapy group. As of August 8, 2023, the median duration of follow-up for survival was 17.2 months. Progression-free survival was longer in the enfortumab vedotin-pembrolizumab group than in the chemotherapy group (median, 12.5 months vs. 6.3 months; hazard ratio for disease progression or death, 0.45; 95% confidence interval [CI], 0.38 to 0.54; P<0.001), as was overall survival (median, 31.5 months vs. 16.1 months; hazard ratio for death, 0.47; 95% CI, 0.38 to 0.58; P<0.001). The median number of cycles was 12 (range, 1 to 46) in the enfortumab vedotin-pembrolizumab group and 6 (range, 1 to 6) in the chemotherapy group. Treatment-related adverse events of grade 3 or higher occurred in 55.9% of the patients in the enfortumab vedotin-pembrolizumab group and in 69.5% of those in the chemotherapy group., Conclusions: Treatment with enfortumab vedotin and pembrolizumab resulted in significantly better outcomes than chemotherapy in patients with untreated locally advanced or metastatic urothelial carcinoma, with a safety profile consistent with that in previous reports. (Funded by Astellas Pharma US and others; EV-302 ClinicalTrials.gov number, NCT04223856.)., (Copyright © 2024 Massachusetts Medical Society.)

Predicting a person's chronological age (CA) from visible skin features using artificial intelligence (AI) is now commonplace. Often, convolutional neural network (CNN) models are built using images of the face as biometric data. However, hands hold telltale signs of a person's age. To determine the utility of using only hand images in predicting CA, we developed two deep CNNs based on 1) dorsal hand images (H) and 2) frontal face images (F). Subjects (n = 1454) were Indian women, 20-80 years, across three geographic cohorts (Mumbai, New Delhi and Bangalore) and having a broad variation in skin tones. Images were randomised: 70% of F and 70% of H were used to train CNNs. The remaining 30% of F and H were retained for validation. CNN validation showed mean absolute error for predicting CA using F and H of 4.1 and 4.7 years, respectively. In both cases correlations of predicted and actual age were statistically significant (r(F) = 0.93, r(H) = 0.90). The CNNs for F and H were validated for dark and light skin tones. Finally, by blurring or accentuating visible features on specific regions of the hand and face, we identified those features that contributed to the CNN models. For the face, areas of the inner eye corner and around the mouth were most important for age prediction. For the hands, knuckle texture was a key driver for age prediction. Collectively, for AI estimates of CA, CNNs based solely on hand images are a viable alternative and comparable to CNNs based on facial images., (© 2024 The Authors. Experimental Dermatology published by John Wiley & Sons Ltd.)

The present study quantifies the environmental and sustainability impacts associated with municipal solid waste management (MSWM) in India which plays a vital environmental issue in recent times. The upsurge in population has resulted in massive waste generation, leading to a concerning rise in the level of greenhouse gas (GHG) emissions. Therefore, the sustainable management of MSW has been discussed and highlights the conversion of MSW into refuse-derived fuel (RDF) to identify its potential for generating electricity in waste-to-energy (WtE) plants. The life cycle assessment (LCA) study has been done to identify and compare the environmental impacts associated with different scenarios (SC) as SC1: landfilling without energy recovery, SC2: open burning and SC3: processing of RDF in WtE plant by considering the nine impact categories from the inventory data obtained over a period of 12 consecutive months (Jan 2021-Jan 2022). The results exhibited that the global warming potential caused by emissions of GHG are in the order of SC1 (1188 kg CO 2 eq) > SC2 (752 kg CO 2 eq) > SC3 (332 kg CO 2 eq), respectively from 1 t of MSW. It is concluded that the WtE plant can help in the reduction of environmental issues, strengthening the capacity of electricity generation and improving the aesthetic view of the city which is socially acceptable as well. Thus, WtE technology can help in achieving sustainable development goal 12 to regenerate the sustainable secondary resources for the twenty-first century and minimize global climate change., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Silver cluster-based solids have garnered considerable attention owing to their tunable luminescence behavior. While surface modification has enabled the construction of stable silver clusters, controlling interactions among clusters at the molecular level has been challenging due to their tendency to aggregate. Judicious choice of stabilizing ligands becomes pivotal in crafting a desired assembly. However, detailed photophysical behavior as a function of their cluster packing remained unexplored. Here, we modulate the packing pattern of Ag 12 clusters by varying the nitrogen-based ligand. CAM-1 formed through coordination of the tritopic linker molecule and NC-1 with monodentate pyridine ligand; established via non-covalent interactions. Both the assemblies show ligand-to-metal-metal charge transfer (LMMCT) based cluster-centered emission band(s). Temperature-dependent photoluminescence spectra exhibit blue shifts at higher temperatures, which is attributed to the extent of the thermal reverse population of the S 1 state from the closely spaced T 1 state. The difference in the energy gap (ΔE ST ) dictated by their assemblies played a pivotal role in the way that Ag 12 cluster assembly in CAM-1 manifests a wider ΔE ST and thus requires higher temperatures for reverse intersystem crossing (RISC) than assembly of NC-1. Such assembly-defined photoluminescence properties underscore the potential toolkit to design new cluster- assemblies with tailored optoelectronic properties., (© 2023 Wiley-VCH GmbH.)

The 17 th Workshop on Recent Issues in Bioanalysis (17 th WRIB) took place in Orlando, FL, USA on June 19-23, 2023. Over 1000 professionals representing pharma/biotech companies, CROs, and multiple regulatory agencies convened to actively discuss the most current topics of interest in bioanalysis. The 17 th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week to allow an exhaustive and thorough coverage of all major issues in bioanalysis of biomarkers, immunogenicity, gene therapy, cell therapy and vaccines.Moreover, in-depth workshops on "EU IVDR 2017/746 Implementation and impact for the Global Biomarker Community: How to Comply with this NEW Regulation" and on "US FDA/OSIS Remote Regulatory Assessments (RRAs)" were the special features of the 17 th edition.As in previous years, WRIB continued to gather a wide diversity of international, industry opinion leaders and regulatory authority experts working on both small and large molecules as well as gene, cell therapies and vaccines to facilitate sharing and discussions focused on improving quality, increasing regulatory compliance, and achieving scientific excellence on bioanalytical issues.This 2023 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2023 edition of this comprehensive White Paper has been divided into three parts for editorial reasons.This publication covers the recommendations on Mass Spectrometry Assays, Regulated Bioanalysis/BMV (Part 1A) and Regulatory Inputs (Part 1B). Part 2 (Biomarkers, IVD/CDx, LBA and Cell-Based Assays) and Part 3 (Gene Therapy, Cell therapy, Vaccines and Biotherapeutics Immunogenicity) are published in volume 16 of Bioanalysis, issues 7 and 8 (2024), respectively.

Background and Purpose: The mainstay of treatment for COVID-19-associated mucormycosis was liposomal Amphotericin B. Other antifungal agents, such as posaconazole and isavuconazole, were used as well. The Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing recommend broth microdilution methods for antifungal susceptibility testing. In this regard, the present study aimed to see what potency and zone diameters correlate with the gold standard broth microdilution method., Materials and Methods: All the isolates were identified by matrix-assisted laser desorption ionization-time-of-flight. In total, 127 isolates of 83  Rhizopus oryzae complex and 44 isolates of  Rhizopus microsporus  complex were selected. Anti-fungal susceptibility testing by disc diffusion and E-test was performed on Mueller Hinton Agar and compared with the CLSI broth microdilution method of Anti-fungal susceptibility testing., Results: Percentage agreement was found to be more in the case of the E test than the disc diffusion method. In the case of R. oryzae , posaconazole had 98.79% agreement with broth microdilution followed by Isavuconazole (97.59%), Itraconazole (96.38%), and Amphotericin B (91.56%)., Conclusion: Disc diffusion correlates well with broth microdilution, although its correlation is weaker when compared to the E test. Effective concentration of Amphotericin B discs for antifungal susceptibility testing depends on the specific Rhizopus species., Competing Interests: The authors declare that there are no financial conflicts of interest to disclose., (Copyright: © 2021, Published by Mazandaran University of Medical Sciences on behalf of Iranian Society of Medical Mycology and Invasive Fungi Research Center.)

Purpose: Erdafitinib is the only FDA-approved targeted therapy for FGFR2/3-altered metastatic urothelial cancer. We characterized the genetic landscape of FGFR-altered urothelial carcinoma and real-world clinical outcomes with erdafitinib, including on-treatment genomic evolution., Experimental Design: Prospectively collected clinical data were integrated with institutional genomic data to define the landscape of FGFR2/3-altered urothelial carcinoma. To identify mechanisms of erdafitinib resistance, a subset of patients underwent prospective cell-free (cf) DNA assessment., Results: FGFR3 alterations predictive of erdafitinib sensitivity were identified in 39% (199/504) of patients with non-muscle invasive, 14% (75/526) with muscle-invasive, 43% (81/187) with localized upper tract, and 26% (59/228) with metastatic specimens. One patient had a potentially sensitizing FGFR2 fusion. Among 27 FGFR3-altered cases with a primary tumor and metachronous metastasis, 7 paired specimens (26%) displayed discordant FGFR3 status. Erdafitinib achieved a response rate of 40% but median progression-free and overall survival of only 2.8 and 6.6 months, respectively (n = 32). Dose reductions (38%, 12/32) and interruptions (50%, 16/32) were common. Putative resistance mutations detected in cfDNA involved TP53 (n = 5), AKT1 (n = 1), and second-site FGFR3 mutations (n = 2)., Conclusions: FGFR3 mutations are common in urothelial carcinoma, whereas FGFR2 alterations are rare. Discordance of FGFR3 mutational status between primary and metastatic tumors occurs frequently and raises concern over sequencing archival primary tumors to guide patient selection for erdafitinib therapy. Erdafitinib responses were typically brief and dosing was limited by toxicity. FGFR3, AKT1, and TP53 mutations detected in cfDNA represent putative mechanisms of acquired erdafitinib resistance., (©2023 American Association for Cancer Research.)

As predictive biomarkers of response to immune checkpoint inhibitors (ICIs) remain a major unmet clinical need in patients with urothelial carcinoma (UC), we sought to identify tissue-based immune biomarkers of clinical benefit to ICIs using multiplex immunofluorescence and to integrate these findings with previously identified peripheral blood biomarkers of response. Fifty-five pretreatment and 12 paired on-treatment UC specimens were identified from patients treated with nivolumab with or without ipilimumab. Whole tissue sections were stained with a 12-plex mIF panel, including CD8, PD-1/CD279, PD-L1/CD274, CD68, CD3, CD4, FoxP3, TCF1/7, Ki67, LAG-3, MHC-II/HLA-DR, and pancytokeratin+SOX10 to identify over three million cells. Immune tissue densities were compared to progression-free survival (PFS) and best overall response (BOR) by RECIST version 1.1. Correlation coefficients were calculated between tissue-based and circulating immune populations. The frequency of intratumoral CD3 + LAG-3 + cells was higher in responders compared to nonresponders (p = 0.0001). LAG-3 + cellular aggregates were associated with response, including CD3 + LAG-3 + in proximity to CD3 + (p = 0.01). Exploratory multivariate modeling showed an association between intratumoral CD3 + LAG-3 + cells and improved PFS independent of prognostic clinical factors (log HR -7.0; 95% confidence interval [CI] -12.7 to -1.4), as well as established biomarkers predictive of ICI response (log HR -5.0; 95% CI -9.8 to -0.2). Intratumoral LAG-3 + immune cell populations warrant further study as a predictive biomarker of clinical benefit to ICIs. Differences in LAG-3 + lymphocyte populations across the intratumoral and peripheral compartments may provide complementary information that could inform the future development of multimodal composite biomarkers of ICI response. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland., (© 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.)

Background and objective Mucormycosis is an emerging and serious angioinvasive infection caused by filamentous fungi related to the order of Mucorales and the class of Mucormycetes. There was a marked increase in the number of cases of mucormycosis in India following the second wave of the coronavirus disease 2019 (COVID-19) pandemic in the year 2021. In this study, we aimed to compare potassium hydroxide (KOH) microscopy with culture for the detection of post-COVID-19 rhino-orbital-cerebral mucormycosis (ROCM). Materials and methods The KOH microscopy was performed with a 10% or 20% KOH-mounted slide with specimens collected from suspected cases of post-COVID-19 ROCM. Simultaneously, the culture was done on Sabouraud dextrose agar (SDA). These were incubated at 37 ℃ and 25 ℃ for 28 days. Diagnostic parameters were calculated by comparing KOH with gold standard culture. Results KOH mount was positive for broad aseptate fungal hyphae in 322 (54.1%) cases, while it was negative in 244 (41.0%) cases. KOH mount was positive for other fungi in 29 (4.8%) samples. The diagnostic accuracy of the KOH mount for Mucorales was 70.3%. KOH mount had a sensitivity of 84.9%, specificity of 61.5%, positive predictive value (PPV) of 56.9%, and negative predictive value (NPV) of 87.2%. Conclusions Based on our findings, the KOH microscopy positivity rate was higher in tissue samples compared to nasal swabs, with a sensitivity of 84.9%, specificity of 61.5%, PPV of 56.9%, and NPV of 87.2%. The overall diagnostic accuracy of the KOH mount for Mucorales was 70.3%., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Hasan et al.)

Purpose: We present the results of a post hoc tumor tissue analysis from the phase 3 MILO/ENGOT-ov11 study (NCT01849874)., Patients and Methods: Mutation/copy-number analysis was performed on tissue obtained pre-randomization. The Kaplan-Meier method was used to estimate progression-free survival (PFS). Unbiased univariate analysis, Cox regression, and binary logistic regression were used to test associations between mutation status and outcomes, including PFS and binary response by local RECIST 1.1., Results: MILO/ENGOT-ov11 enrolled 341 patients, ranging in age from 22 to 79, from June, 2013 to April, 2016. Patients were randomized 2:1 to binimetinib or physician's choice of chemotherapy (PCC). The most commonly altered gene was KRAS (33%). In 135 patients treated with binimetinib with response rate (RR) data, other detected MAPK pathway alterations included: NRAS (n = 11, 8.1%), BRAF V600E (n = 8, 5.9%), RAF1 (n = 2, 1.5%), and NF1 (n = 7, 5.2%). In those with and without MAPK pathway alterations, the RRs with binimetinib were 41% and 13%, respectively. PFS was significantly longer in patients with, compared with those without, MAPK pathway alterations treated with binimetinib [HR, 0.5; 95% confidence interval (CI) 0.31-0.79]. There was a nonsignificant trend toward PFS improvement in PCC-treated patients with MAPK pathway alterations compared with those without (HR, 0.82; 95% CI, 0.43-1.59)., Conclusions: Although this hypothesis-generating analysis is limited by multiple testing, higher RRs and longer PFS were seen in patients with low-grade serous ovarian cancer (LGSOC) treated with binimetinib, and to a lesser extent in those treated with PCC, who harbored MAPK pathway alterations. Somatic tumor testing should be routinely considered in patients with LGSOC and used as a future stratification factor., (©2023 The Authors; Published by the American Association for Cancer Research.)

Background: Sudden upsurge in cases of COVID-19 Associated Mucormycosis (CAM) following the second wave of the COVID-19 pandemic was recorded in India. This study describes the clinical characteristics, management and outcomes of CAM cases, and factors associated with mortality., Methods: Microbiologically confirmed CAM cases were enrolled from April 2021 to September 2021 from ten diverse geographical locations in India. Data were collected using a structured questionnaire and entered into a web portal designed specifically for this investigation. Bivariate analyses and logistic regression were conducted using R version 4.0.2., Results: A total of 336 CAM patients were enrolled; the majority were male (n = 232, 69.1%), literate (n = 261, 77.7%), and employed (n = 224, 66.7%). The commonest presenting symptoms in our cohort of patients were oro-facial and ophthalmological in nature. The median (Interquartile Range; IQR) interval between COVID diagnosis and admission due to mucormycosis was 31 (18, 47) days, whereas the median duration of symptoms of CAM before hospitalization was 10 (5, 20) days. All CAM cases received antifungal treatment, and debridement (either surgical or endoscopic or both) was carried out in the majority of them (326, 97.02%). Twenty-three (6.9%) of the enrolled CAM cases expired. The odds of death in CAM patients increased with an increase in HbA1c level (aOR: 1.34, 95%CI: 1.05, 1.72) following adjustment for age, gender, education and employment status., Conclusion: A longer vigil of around 4-6 weeks post-COVID-19 diagnosis is suggested for earlier diagnosis of CAM. Better glycemic control may avert mortality in admitted CAM cases., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: FGFR alterations are reported across various malignancies and might act as oncogenic drivers in multiple histologies. Erdafitinib is an oral, selective pan-FGFR tyrosine kinase inhibitor with activity in FGFR-altered advanced urothelial carcinoma. We aimed to evaluate the safety and activity of erdafitinib in previously treated patients with FGFR-altered advanced solid tumours., Methods: The single-arm, phase 2 RAGNAR study was conducted at 156 investigative centres (hospitals or oncology practices that are qualified oncology study centres) across 15 countries. The study consisted of four cohorts based on tumour histology and patient age; the results reported in this Article are for the primary cohort of the study, defined as the Broad Panel Cohort, which was histology-agnostic. We recruited patients aged 12 years or older with advanced or metastatic tumours of any histology (except urothelial cancer) with predefined FGFR1-4 alterations (mutations or fusions according to local or central testing). Eligible patients had disease progression on at least one previous line of systemic therapy and no alternative standard therapy available to them, and an Eastern Cooperative Oncology Group performance status of 0-1 (or equivalent for adolescents aged 12-17 years). Patients received once-daily oral erdafitinib (8 mg/day with provision for pharmacodynamically guided up-titration to 9 mg/day) on a continuous 21-day cycle until disease progression or intolerable toxicity. The primary endpoint was objective response rate by independent review committee according to Response Evaluation Criteria In Solid Tumors (RECIST), version 1.1, or Response Assessment In Neuro-Oncology (RANO). The primary analysis was conducted on the treated population of the Broad Panel Cohort. This ongoing study is registered with ClinicalTrials.gov, number NCT04083976., Findings: Patients were recruited between Dec 5, 2019, and Feb 15, 2022. Of 217 patients treated with erdafitinib, 97 (45%) patients were female and 120 (55%) were male. The data cutoff was Aug 15, 2022. At a median follow-up of 17·9 months (IQR 13·6-23·9), an objective response was observed in 64 (30% [95% CI 24-36]) of 217 patients across 16 distinct tumour types. The most common grade 3 or higher treatment-emergent adverse events related to erdafitinib were stomatitis (25 [12%]), palmar-plantar erythrodysaesthesia syndrome (12 [6%]), and hyperphosphataemia (11 [5%]). The most commonly occurring serious treatment-related adverse events (grade 3 or higher) were stomatitis in four (2%) patients and diarrhoea in two (1%). There were no treatment-related deaths., Interpretation: RAGNAR results show clinical benefit for erdafitinib in the tumour-agnostic setting in patients with advanced solid tumours with susceptible FGFR alterations who have exhausted other treatment options. These results support the continued development of FGFR inhibitors in patients with advanced solid tumours., Funding: Janssen Research & Development., Competing Interests: Declaration of interests SP has received consulting fees from Ipsen, Janssen, Novartis, and Zymeworks; and institutional research funding from 4D Pharma, Arcus Biosciences, Astellas Pharma, Boehringer Ingelheim, Bristol Myers Squibb, Elicio Therapeutics, Janssen, Lilly, Mirati Therapeutics, NGM Biopharmaceuticals, Novartis, Purple Biotech, Rgenix, and Xencor. MS has received consulting fees from Amgen, AstraZeneca, Boehringer Ingelheim, Bristol Myers Squibb, GlaxoSmithKline, Janssen Oncology, Merck Serono, Novartis, Roche, Sanofi, and Takeda; honoraria from Amgen, Boehringer Ingelheim, Bristol Myers Squibb, Janssen-Cilag, and Novartis; institutional research funding from AstraZeneca and Bristol Myers Squibb; and has institutional patents, royalties, or other intellectual property for a highly sensitive method for mutation detection by PCR. GI has received consulting fees from Basilea, Bayer, Flare Therapeutics, Janssen, Loxo/Lilly, and Mirati Therapeutics; speakers’ bureau fees from Gilead Sciences and Lynx Group; and institutional research funding from Bayer, Debiopharm Group, Janssen, Mirati Therapeutics, Novartis, and Seagen. OW has received consulting fees from Bayer US, LCC, Bristol Myers Squibb, Day One Therapeutics, Novartis, and Roche; and research funding from AstraZeneca, Bayer US, LLC, Blueprint Medicines, Day One Therapeutics, GlaxoSmithKline, Janssen, Lilly, Loxo, Novartis, and Roche. TD has received consulting fees from AbbVie, Amgen, Boehringer Ingelheim, Chugai Pharma, Daiichi Sankyo, Janssen, Kaken Pharmaceutical, Kyowa Kirin, MSD, Otsuka, PRA Health Science, Rakuten Medical, Shionogi, Sumitomo Dainippon Pharma, Taiho Pharmaceutical, and Takeda; honoraria from AstraZeneca, Bristol Myers Squibb Japan, Chugai Pharma, Daicchi Sankyo, Ono Pharmaceutical, Rakuten Medical Japan, and Taiho Pharmaceutical; and institutional research funding from AbbVie, Boehringer Ingelheim, Bristol Myers Squibb, Chugai Pharma, Daiichi Sankyo, Eisai, IQVIA, Janssen, MSD, Ono Pharmaceutical, Novartis, Pfizer, PRA Health Sciences, Sumitomo Dainippon Pharma, and Taiho Pharmaceutical. JT has received consulting fees from Array BioPharma, AstraZeneca, Bayer, Boehringer Ingelheim, Cardiff Oncology, Chugai Pharma, Daiichi Sankyo, F Hoffman LaRoche, Genentech, HalioDx, Ikena Oncology, Inspirna, IQVIA, Lilly, Menarini, Merck Serono, Merus, Mirati Therapeutics, MSD, NeoPhore, Ona Therapeutics, Orion Biotechnology, Hutchison MediPharma, Novartis, Peptomyc, Pfizer, Pierre Fabre, Samsung Bioepis, Sanofi, Scandion Oncology, Scorpion Therapeutics, Servier, Sotio, Taiho Pharmaceutical, Tessa Therapeutics, TheraMyc, and Tolremo; other financial interests from Imedex/HMP, Medscape, MJH Life Sciences, Peerview, Physicians’ Education Resource; and stock and other ownership interests from Oniria Therapeutics. DAR has received consulting fees from Advantagene, Agenus, Agios, AnHeart Therapeutics, Bayer, Bristol Myers Squibb, Delmar Pharmaceuticals, Ellipses Pharma, EMD Serono, Genenta Science, Imvax, Kintara Therapeutics, Kiyatec, Medicenna, Merck, Merck KGaA, Novocure, Oncorus, Regeneron, Taiho Pharmaceutical, and Vivacitas Oncology; honoraria from Advantagene, Agenus, AnHeart Therapeutics, Bayer, Bristol Myers Squibb, Deciphera, DelMar Pharmaceuticals, Ellipses Pharma, EMD Serono, Genenta Science, Imvax, Inovio Pharmaceuticals, Kintara Therapeutics, Kiyatec, Medicenna, Merck, Merck KGaA, Neuvogen, Oncorus, Novocure, Regeneron, Sumitomo Dainippon Pharma, Taiho Pharmaceutical, Vivacitas Oncology, and Y-mAbs Therapeutics; and institutional research funding from Acerta Pharma, Agenus, Celldex, EMD Serono, Enterome, Incyte, and Omniox. CM has received consulting fees from Amgen, Astellas Pharma, AstraZeneca, Bayer, BeiGene, Blueprint Medicines, Bristol Myers Squibb, Celgene, Debiopharm Group, Faron Pharmaceuticals, Genentech/Roche, Innate Pharma, Ipsen, Janssen, Lilly, MSD, Novartis, Orion, Pfizer, PharmaMar, Sanofi, and Taiho Pharmaceutical. AM has received consulting fees from Bayer, Chugai, GlaxoSmithKline, Janssen, Merck, and Novartis Oncology; has been reimbursed for travel, accommodations, or expenses from Amgen; and has received fees for participating in a data safety monitoring board or advisory board from Faron, Genmab, Janssen, Merck, and Takeda. IL has received consulting fees from Boehringer Ingelheim; honoraria from Agenus, Amgen, AstraZeneca, BeiGene, Bristol Myers Squibb, Celon, Cullinan Oncology, Jacobio, Janssen, Loxo, Macrogenics, Menarini, MSD, Pfizer, Rhizen, Roche, Sanofi, and Takeda; institutional research funding from Agenus and Roche; and has been reimbursed for travel, accommodations, or expenses from Bristol Myers Squibb. OC has received consulting fees from Bayer US, LLC, and Janssen; honoraria from Bayer US, LLC, Janssen, Merck, Novartis, and Pfizer; and speakers’ bureau fees from Merck and Pfizer. DA has received consulting fees from AstraZeneca, Boston Scientific, Bristol Myers Squibb, CRA International, Gilead, Janssen Cilag, MSD, Onkowissen, Pierre Fabre Pharma, Seagen, and Terumo; honoraria from Amgen, Aptitude Health, Art Tempi Media, AstraZeneca, Boehringer Ingelheim, Boston Scientific, Bristol Myers Squibb, Clinical Care Options, Eisai, From Research to Practice, GlaxoSmithKline, Imedex, Ipsen, MCI, MedAhead (Austria), Merck Serono, MSD, Pierre Fabre Pharma, PRMA Consulting, Roche, Sanofi (Genzyme), Seagen, Servier, Streamitup Germany, Tactics MD LLC, Terumo, Viatris, and WebMD; fees for editorial roles with Elsevier; fees and an institutional education grant from AbbVie; institutional trial support from Bristol Myers Squibb and Oncolytics; and has served in a leadership role with EORTC. MG has received consulting fees from Celularity and Guardant. HW has received honoraria from Bristol Myers Squibb, GlaxoSmithKline, and Novartis; and has been reimbursed for travel, accommodations, or expenses from Janssen. KS, LC, SN, CH, STh, AS-W, STr, and HS received personal fees from Janssen during the conduct of the study. YL has received consulting fees from Astellas Pharma, AstraZeneca, Bristol Myers Squibb, Immunomedics, Janssen (personal and institutional), MSD Oncology (personal and institutional), Loxo/Lilly, Pfizer/EMD Serono, Roche, and Taiho Pharmaceutical; has been reimbursed for travel, accommodations, or expenses from Astellas, AstraZeneca, Janssen Oncology, MSD Oncology, and Roche; and has received institutional research funding from Astellas Pharma, AstraZeneca, Basilea, Bristol Myers Squibb, Exelixis, Gilead Sciences, Incyte, Janssen Oncology, Merck KGaA, MSD Oncology, Nektar, Pfizer, Roche, Sanofi, and Taiho Pharmaceutical. SQ declares no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The morbidity associated with radical cystectomy (RC) for muscle-invasive bladder cancer (MIBC) has fueled investigations into the feasibility of bladder preservation strategies after a favorable clinical response to neoadjuvant therapy (NAT). Identifying optimal candidates for bladder preservation is predicated on our ability to identify tumors with inherent cisplatin sensitivity and accurately stage patients before and after NAT. In the present review, we evaluate the accuracy and limitations of contemporary staging modalities and investigate clinical outcomes in patients with MIBC who were managed with bladder preservation after NAT. Lastly, we discuss the predictive role of cisplatin-sensitizing DNA damage response (DDR) gene alterations as a foundational component to current prospective clinical trials evaluating bladder preservation in the setting of MIBC., Competing Interests: Conflict of Interest The authors report no conflicts of interest pertinent to the reviewed material., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Various radiotherapeutic regimens are used in the treatment of bladder cancer., Objective: We aimed to evaluate early toxicity and outcomes associated with hypofractionated radiation therapy (Hypo-RT), 55Gy in 20 fractions., Material and Methods: We identified 40 patients who received definitive Hypo-RT for localized bladder cancer. Most patients were men (62.5%), elderly (median age 82), had high Charlson Comorbidity Index score (median 7, range 4-9) and were nonsurgical candidates (80%). Sixty-eight percent had a macroscopically complete transurethral resection of bladder tumor (TURBT) and 33 patients (82.5%) received concurrent chemotherapy. Acute (< =3mo) and late (>3mo) toxicities were assessed according to CTCAE v4.0. Survival outcomes were estimated using the Kaplan-Meier method. Median follow up after Hypo-RT was 32 months (95% CI: 28-49 months)., Results: Overall rates of acute grade 2 genitourinary (GU) and gastrointestinal (GI) toxicities were 40% each, most commonly urinary frequency and diarrhea. Two cases of acute grade 3 GU/GI toxicity occurred. Late grade 2+ toxicity occurred in 3 patients (7.5%): 2 grade 2 GU and 1 grade 3 GI. Seventy-seven percent achieved a complete response (CR). Six patients (20%) developed disease recurrence at a median time of 9.1 months. The estimated 2-year DFS and 2-year DSS rate were 59% (95% CI, 45-78%) and 78% (95% CI, 65-93%), respectively. Receipt of concurrent chemotherapy ( p  = 0.003) and achieving a CR ( p  = 0.018) were univariably associated with improved DSS. Tis component was associated with worse DSS ( p  = 0.015)., Conclusion: Hypo-RT had a favorable toxicity profile and encouraging cancer control outcomes in this mostly elderly and frail patient cohort., Competing Interests: Jonathan E. Rosenberg and Bernard Bochner are Editorial Board Members of this journal, but were not involved in the peer-review process nor had access to any information regarding its peer-review. Assaf Moore, Stephanie M. Lobaugh, Zhigang Zhang, Gopa Iyer, Min Yuen Teo, Timothy Donahue, David Aramburu Nunez, Alexandra Dreyfuss, Daniel Gorovets, Michael J. Zelefsky and Marisa A. Kollmeier report no perceived conflicts of interest relevant to the current work., (© 2023 – The authors. Published by IOS Press.)

Purpose: Neoadjuvant chemotherapy (NAC) has proven survival benefits for patients with invasive urothelial carcinoma of the bladder, yet its role for upper tract urothelial carcinoma (UTUC) remains undefined. We conducted a multicenter, single-arm, phase II trial of NAC with gemcitabine and split-dose cisplatin (GC) for patients with high-risk UTUC before extirpative surgery to evaluate response, survival, and tolerability., Methods: Eligible patients with defined criteria for high-risk localized UTUC received four cycles of split-dose GC before surgical resection and lymph node dissection. The primary study end point was rate of pathologic response (defined as < ypT2N0). Secondary end points included progression-free survival (PFS), overall survival (OS), and safety and tolerability., Results: Among 57 patients evaluated, 36 (63%) demonstrated pathologic response (95% CI, 49 to 76). A complete pathologic response (ypT0N0) was noted in 11 patients (19%). Fifty-one patients (89%) tolerated at least three complete cycles of split-dose GC, 27 patients (47%) tolerated four complete cycles, and all patients proceeded to surgery. With a median follow up of 3.1 years, 2- and 5-year PFS rates were 89% (95% CI, 81 to 98) and 72% (95% CI, 59 to 87), while 2- and 5-year OS rates were 93% (95% CI, 86 to 100) and 79% (95% CI, 67 to 94), respectively. Pathologic complete and partial responses were associated with improved PFS and OS compared with nonresponders (≥ ypT2N any; 2-year PFS 100% and 95% v 76%, P < .001; 2-year OS 100% and 100% v 80%, P < .001)., Conclusion: NAC with split-dose GC for high-risk UTUC is a well-tolerated, effective therapy demonstrating evidence of pathologic response that is associated with favorable survival outcomes. Given that these survival outcomes are superior to historical series, these data support the use of NAC as a standard of care for high-risk UTUC, and split-dose GC is a viable option for NAC.

Bladder cancer is an aggressive and lethal disease. Even when presenting as localized muscle-invasive disease, the 5-year survival rate is about 70%, and the recurrence rate after radical cystectomy is approximately 50%. Neoadjuvant chemotherapy (NAC) has the potential to downstage the primary tumor and treat micrometastases, leading to a decrease in recurrence rates and an increase in cure rates. There is level 1 evidence in favor of neoadjuvant cisplatin-based chemotherapy prior to radical cystectomy. However, data from clinical trials evaluating NAC for patients undergoing bladder-sparing treatments are less robust, so this strategy remains controversial. The response to NAC is prognostic and patients with favorable pathological response have better overall survival. Strategies to select patients based on molecular biomarkers have the potential to guide treatment decisions and even de-intensify treatment, avoiding local treatment for those with complete responses to systemic therapy. This review outlines the current literature on the use of NAC in the context of bladder preservation for muscle-invasive bladder cancer, highlights neoadjuvant studies in patients ineligible for cisplatin-based NAC, and discusses novel bladder-preservation strategies, including multimodality combinations and biomarker-driven studies of definitive chemotherapy., (Published by Elsevier Inc.)

Migration is often understood to be a livelihood strategy to cope with the effects of environmental threats and climate change. Yet, the extent to which migration decisions differ due to the type, severity, and frequency of environmental events has been little explored. This paper employs household surveys in southwestern Bangladesh to explore this research gap. A multinominal regression model is used to simulate reported future migration decisions (200 sample households) in the context of both rapid-onset (i.e. cyclone and flood) and slow-onset (salinity, siltation, and riverbank erosion) environmental phenomena. Results show: i) previous disaster experience and increasing conflict in the community motivate migration in the near future in the context of slow-onset phenomena (salinity); (ii) economic strength and self-efficacy increase non-migration intention in both contexts of sudden and slow-onset events; and (iii) the extent and pattern of these influences on migration differ across demographics, including education, religion, and age. Importantly, this analysis shows that the relationship between migration decisions and the type, severity, and frequency of environmental events is influenced by socioeconomic conditions. Therefore, this research supports future adaptation planning specifically tailored to the type and exposure of extreme environmental events., Competing Interests: Disclosure statement No potential conflict of interest was reported by the author(s).

Precision oncology relies on the accurate molecular characterization of individual patients with cancer at the time of treatment initiation. However, tumor molecular profiles are not static, and cancers continually evolve because of ongoing mutagenesis and clonal selection. Here, we performed genomic analyses of primary tumors, metastases, and plasma collected from individual patients to define the concordance of actionable genomic alterations and to identify drivers of metastatic disease progression. We observed a high degree of discordance of actionable genomic alterations, with 23% discordant between primary and metastatic disease sites. Among chromatin-modifying genes, ARID1A mutations, when discordant, were exclusive to the metastatic tumor samples. Our findings indicate that the high degree of lesion-to-lesion genomic heterogeneity may be a barrier to precision oncology approaches for bladder cancer and that circulating tumor DNA profiling may be preferred to tumor sequencing for a subset of patients., Competing Interests: Author contributions Conceptualization, T.N.C., Z.C., H.W., S.C., H.A.A.-A., and D.B.S; methodology, T.N.C., Z.C., H.W., A.T.L., S.C., M.T.A.D., N.A., S.D., J.E.R., N.V., E.J.P., S.P.G., W.H., H.A.A.-A., and D.B.S.; investigation, T.N.C., Z.C., H.W., A.T.L., S.C., M.T.A.D., N.A., C.C., S.D., J.R., N.V., S.P.G., W.H., and H.A.A.-A.; resources, P.R., J.R., D.F.B., M.Y.T., B.H.B., M.B., E.J.P., G.I., H.A.A.-A., and D.B.S.; formal analysis, T.N.C., Z.C., A.T.L., S.C., M.T.A.D., I.O., and W.H.; writing, T.N.C., Z.C., A.T.L., and D.B.S.; review & editing, T.N.C., Z.C., H.A.A.-A., and D.B.S.; supervision, D.B.S. Declaration of interests D.B.S. has served as a consultant for/received honorarium from Pfizer, Loxo/Lilly Oncology, Vividion Therapeutics, Scorpion Therapeutics, Fore Therapeutics, FOG Pharma, Rain Therapeutics, and BridgeBio. H.A.A.-A. provided consultation to AstraZeneca, Janssen Biotech, Bristol-Myers-Squibb, and Paige.ai. J.E.R. has served as a consultant for Astellas, Seagen, Merck, Roche, Genentech, AstraZeneca, Janssen Biotech, Gilead, Pfizer, EMD-Serono, Mirati, Boehringer Ingelheim, Pharmacyclis, GSK, Infinity, Tyra BioSciences, Bayer, and QED Therapeutics and received honoraria from EMD-Serono. M.B. has served as a consultant for Eli Lilly and PetDx and has a patent pending on “Systems and Methods for Detecting Cancer Via cfDNA Screening.” S.D. has served as a consultant for Roche., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Urothelial Cancer - Genomic Analysis to Improve Patient Outcomes and Research (NCT02643043), UC-GENOME, is a genomic analysis and biospecimen repository study in 218 patients with metastatic urothelial carcinoma. Here we report on the primary outcome of the UC-GENOME-the proportion of subjects who received next generation sequencing (NGS) with treatment options-and present the initial genomic analyses and clinical correlates. 69.3% of subjects had potential treatment options, however only 5.0% received therapy based on NGS. We found an increased frequency of TP53 E285K mutations as compared to non-metastatic cohorts and identified features associated with benefit to chemotherapy and immune checkpoint inhibition, including: Ba/Sq and Stroma-rich subtypes, APOBEC mutational signature (SBS13), and inflamed tumor immune phenotype. Finally, we derive a computational model incorporating both genomic and clinical features predictive of immune checkpoint inhibitor response. Future work will utilize the biospecimens alongside these foundational analyses toward a better understanding of urothelial carcinoma biology., (© 2022. The Author(s).)

Cancers arising from the bladder urothelium often exhibit lineage plasticity with regions of urothelial carcinoma adjacent to or admixed with regions of divergent histomorphology, most commonly squamous differentiation. To define the biologic basis for and clinical significance of this morphologic heterogeneity, here we perform integrated genomic analyses of mixed histology bladder cancers with separable regions of urothelial and squamous differentiation. We find that squamous differentiation is a marker of intratumoral genomic and immunologic heterogeneity in patients with bladder cancer and a biomarker of intrinsic immunotherapy resistance. Phylogenetic analysis confirms that in all cases the urothelial and squamous regions are derived from a common shared precursor. Despite the presence of marked genomic heterogeneity between co-existent urothelial and squamous differentiated regions, no recurrent genomic alteration exclusive to the urothelial or squamous morphologies is identified. Rather, lineage plasticity in bladder cancers with squamous differentiation is associated with loss of expression of FOXA1, GATA3, and PPARG, transcription factors critical for maintenance of urothelial cell identity. Of clinical significance, lineage plasticity and PD-L1 expression is coordinately dysregulated via FOXA1, with patients exhibiting morphologic heterogeneity pre-treatment significantly less likely to respond to immune checkpoint inhibitors., (© 2022. The Author(s).)

Introduction: Small cell carcinoma of the bladder (SCCB) is a rare variant of bladder cancer with poor outcomes. We evaluated long-term outcomes of nonmetastatic (M0) and metastatic (M1) SCCB and correlated pathologic response with genomic alterations of patients treated with neoadjuvant chemotherapy (NAC)., Patients and Methods: Clinical history and pathology samples from SCCB patients diagnosed at our institution were reviewed., Results: One hundred and ninety-nine SCCB patients were identified. (M0: 147 [74%]; M1: 52 [26%]). Among M0 patients, 108 underwent radical cystectomy (RC) (NAC: 71; RC only: 23; adjuvant chemotherapy: 14); 14 received chemoradiotherapy; the rest received chemotherapy alone or no cancer-directed therapy. RC-only patients had a median follow-up of 9.1 years, and median disease-free survival (DFS) and overall survival (OS) were 1.1 and 1.2 years, respectively. NAC patients had pathologic response (

Floods and river-bank erosion are the most frequent natural hazards in India, specifically in the deltaic regions. In West Bengal, floods and river-bank erosion predominantly affect Malda district as it is located in the moribund part of the Bengal delta. This article studies the recent trend of shifting course of the River Ganga and the effects of floods and consequent river-bank erosion on livelihoods of the residents of chars [The chars (called Diara in the upper reaches of the Gangetic plains) are virgin, low-lying river islands and sand bars occurring in the plains, particularly the deltaic parts of rivers (Lahiri-Dutt and Samanta, South Asia: J South Asia Stud 30:327-350, 2007).] and river-bank areas of Manikchak block in the Malda district. Around 300 sample households were selected by random stratified sampling technique from four gram panchayats of Manikchak block. Both primary and secondary data have been used. After analysing satellite images from the year 1973 to 2018, it has been observed that the River Ganga continues to shift eastwards and is eroding villages one after another. Inhabitants face multidimensional obstacles to run their households. Large numbers of people are displaced every year due to loss of land. Failure in facilitating the required assistance in the form of alternative spaces for resettlement and other disaster-mitigating public support systems against these hazards would make it impossible for the deplorable condition of the vulnerable people to improve., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Nature B.V. 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

The burgeoning noncovalent interactions between π-acidic aromatic surfaces and anions have been recently shown to have unique functional relevance in anion transport, ion sensing, and organocatalysis. Despite its potential to instigate charge-transfer (CT) states, modulation of the emission features by toggling between the excited states using anion-π interactions is not yet explored. On the other hand, excited states with CT characteristics play an important role in the ambient triplet harvesting of organic chromophores. In this context, herein we propose an anion-π-based molecular design for the introduction of emissive singlet and triplet CT excited states, thereby expanding the functional scope of these weak supramolecular interactions. In the present study, we investigate the anion-π-induced emission from the singlet ( 1 CT) and triplet ( 3 CT) CT states of a dibromo dicationic pyromellitic diimide derivative. Remarkably, we accomplish dual room temperature phosphorescence emission from the anion-π-mediated 3 CT state along with the locally excited triplet state ( 3 LE) in solution phase using an organic-inorganic supramolecular scaffolding strategy. Comprehensive steady-state and time-resolved spectroscopy along with theoretical calculations provide detailed insights into the excited-state manifolds of phosphor. We envisage that the present study will expedite new molecular designs based on weak intermolecular interactions for the excited-state engineering of organic chromophores to facilitate ambient triplet harvesting and CT emission.

Purpose: To compare oncologic outcomes and genomic alteration profiles in patients with bladder and urachal adenocarcinoma, urothelial carcinoma (UC) with glandular differentiation, and UC, not otherwise specified (NOS) undergoing surgical resection, with emphasis on response to systemic therapy., Methods: We identified patients with bladder cancer with glandular variants who underwent surgical resection at Memorial Sloan Kettering from 1995 to 2018 (surgical cohort) and/or patients who had tumor sequencing using a targeted next-generation sequencing platform (genomics cohort). Pathologic complete and partial response rates to neoadjuvant chemotherapy (NAC) and recurrence-free and cancer-specific survival were measured. Alteration frequencies between histologic subtypes were compared., Results: Thirty-seven patients with bladder adenocarcinoma, 46 with urachal adenocarcinoma, 84 with UC with glandular differentiation, and 1,049 with UC, NOS comprised the surgical cohort. Despite more advanced disease in patients with bladder and urachal adenocarcinoma, no significant differences in recurrence or cancer-specific survival by histology were observed after adjusting for stage. In patients with UC with glandular differentiation, NAC resulted in partial (≤ pT1N0) and complete (pT0N0) responses in 28% and 17%, respectively. Bladder and urachal adenocarcinoma genomic profiles resembled colorectal adenocarcinoma with frequent TP53 , KRAS , and PIK3CA alterations while the genomic profile of UC with glandular differentiation more closely resembled UC, NOS. Limitations include retrospective nature of analysis and small numbers of nonurothelial histology specimens., Conclusion: The genomic profile of bladder adenocarcinomas resembled colorectal adenocarcinomas, whereas UC with glandular differentiation more closely resembled UC, NOS. Differences in outcomes among patients with glandular bladder cancer variants undergoing surgical resection were largely driven by differences in stage. Cisplatin-based NAC demonstrated activity in UC with glandular differentiation, suggesting NAC should be considered for this histologic variant.

2,2'-Bipyridine based bisphosphine [C 5 H 3 N{N(H)CH 2 PPh 2 }] 2 (1) and its bischalcogenide derivatives [C 5 H 3 N{N(H)CH 2 P(E)Ph 2 }] 2 (2, E = O; 3, E = S; 4, E = Se) were synthesized, and further reacted with BF 3 ·Et 2 O/Et 3 N to form doubly B ← N fused compounds [C 5 H 3 N(BF 2 ){NCH 2 P(E)Ph 2 }] 2 (5, E = O; 6, E = S; 7, E = Se) in excellent yields. The influence of the PE bonds on the electronic properties of the doubly B ← N fused systems and their structural features were investigated in detail, supported by extensive experimental and computational studies. Compound 6 exhibited a very high quantum yield of ϕ = 0.56 in CH 2 Cl 2 , whereas compound 7 showed a least quantum yield of ϕ = 0.003 in acetonitrile. Density functional theory (DFT) calculations demonstrated that the LUMO/HOMO of compounds 5-7 mostly delocalized over the entire π-conjugated frameworks. The involvement of PE bonds in the HOMO energy level of these compounds follows the order: PO < PS < PSe. Time-correlated single photon counting (TCSPC) experiments of compounds 5-7 revealed the singlet lifetime of 4.26 ns for 6, followed by 4.03 ns for 5 and a lowest value of 2.18 ns ( τ 1 ) and 0.47 ns ( τ 2 ) with a double decay profile for 7. Our findings provide important strategies for the design of highly effective B ← N bridged compounds and tuning their photophysical properties by oxidizing phosphorus with different chalcogens. Compounds 5 and 6 have been employed as green emitters ( λ em = 515 nm) in fluorescent organic light-emitting diodes (OLEDs). For compound 5, doped into the poly(9-vinylcarbazole) (PVK) matrix with 5 wt% doping concentration, nearly 90 Cd m -2 luminance with 0.022% external quantum efficiency (EQE) was achieved. The best performance was observed for compound 6 doped into PVK by 1 wt% having a maximum luminance of 350 Cd m -2 and a similar EQE value.

Purpose: Neoadjuvant gemcitabine and cisplatin (GC) followed by radical cystectomy (RC) is standard for patients with muscle-invasive bladder cancer (MIBC). On the basis of the activity of atezolizumab (A) in metastatic BC, we tested neoadjuvant GC plus A for MIBC., Methods: Eligible patients with MIBC (cT2-T4aN0M0) received a dose of A, followed 2 weeks later by GC plus A every 21 days for four cycles followed 3 weeks later by a dose of A before RC. The primary end point was non-muscle-invasive downstaging to < pT2N0., Results: Of 44 enrolled patients, 39 were evaluable. The primary end point was met, with 27 of 39 patients (69%) < pT2N0, including 16 (41%) pT0N0. No patient with < pT2N0 relapsed and four (11%) with ≥ pT2N0 relapsed with a median follow-up of 16.5 months (range: 7.0-33.7 months). One patient refused RC and two developed metastatic disease before RC; all were considered nonresponders. The most common grade 3-4 adverse event (AE) was neutropenia (n = 16; 36%). Grade 3 immune-related AEs occurred in five (11%) patients with two (5%) requiring systemic steroids. The median time from last dose of chemotherapy to surgery was 7.8 weeks (range: 5.1-17 weeks), and no patient failed to undergo RC because of AEs. Four of 39 (10%) patients had programmed death-ligand 1 (PD-L1)-positive tumors and were all < pT2N0. Of the patients with PD-L1 low or negative tumors, 23 of 34 (68%) achieved < pT2N0 and 11 of 34 (32%) were ≥ pT2N0 ( P = .3 for association between PD-L1 and < pT2N0)., Conclusion: Neoadjuvant GC plus A is a promising regimen for MIBC and warrants further study. Patients with < pT2N0 experienced improved relapse-free survival. The PD-L1 positivity rate was low compared with published data, which limits conclusions regarding PD-L1 as a predictive biomarker., Competing Interests: Samuel A. FuntEmployment: ByHeart (I)Stock and Other Ownership Interests: Kite, a Gilead company, Urogen pharma, Allogene Therapeutics, Neogene Therapeutics, Kronos Bio, Vida Ventures, IconOVir BioConsulting or Advisory Role: Merck, ImmunaiResearch Funding: Genentech/Roche (Inst), AstraZeneca (Inst), Decibel Therapeutics (Inst)Travel, Accommodations, Expenses: Bristol Myers Squibb, AstraZeneca/MedImmune Hikmat Al-AhmadieConsulting or Advisory Role: Bristol Myers Squibb, EMD Serono, AstraZeneca/MedImmune, Janssen Biotech, PAIGE.AI Min Yuen TeoConsulting or Advisory Role: Janssen OncologyResearch Funding: Bristol Myers Squibb (Inst), Clovis Oncology (Inst), Pharmacyclics (Inst) Chung-Han LeeHonoraria: Intellisphere, Research to Practice, American Institute of Continuing Medical EducationConsulting or Advisory Role: Eisai, Bristol Myers Squibb, Merck, Pfizer/EMD Serono, ExelixisResearch Funding: Eisai (Inst), Bristol Myers Squibb (Inst), Calithera Biosciences (Inst), Exelixis (Inst), Merck (Inst) David AggenConsulting or Advisory Role: Boehringer Ingelheim, Seattle Genetics, Astellas PharmaPatents, Royalties, Other Intellectual Property: University of Illinois—Urbana Champaign(OPTIONAL) Open Payments Link: https://openpaymentsdata.cms.gov/physician/4226107 Deaglan McHughConsulting or Advisory Role: Progenics Arlyn ApolloEmployment: Covera Health (I)Leadership: Covera Health (I)Stock and Other Ownership Interests: Covera Health (I)Consulting or Advisory Role: Covera Health (I)Travel, Accommodations, Expenses: Covera Health (I) Kaitlyn FranceseConsulting or Advisory Role: Seattle Genetics, Astellas Pharma Yuanquan YangStock and Other Ownership Interests: Bristol Myers Squibb, Pfizer, AstraZenecaConsulting or Advisory Role: The Whiteoak Group Eugene PietzakHonoraria: UpToDateConsulting or Advisory Role: Merck, Chugai Pharma, QED Therapeutics, Janssen Alvin C. GohConsulting or Advisory Role: MedtronicTravel, Accommodations, Expenses: Medtronic William C. HuangHonoraria: Urogen pharmaConsulting or Advisory Role: Intuitive SurgicalResearch Funding: SonaCare Medical, photocure, Storz, Storz, Merck (Inst), Intuitive Surgical (Inst)Travel, Accommodations, Expenses: Photocure Dean F. BajorinConsulting or Advisory Role: Merck, Dragonfly Therapeutics, Fidia Farmaceutici S. p. A, Bristol Myers Squibb FoundationResearch Funding: Novartis (Inst), Merck (Inst), Bristol Myers Squibb (Inst), AstraZeneca (Inst), Astellas Pharma (Inst), Seattle Genetics/Astellas (Inst)Travel, Accommodations, Expenses: Merck Gopa IyerConsulting or Advisory Role: Bayer, Janssen, Mirati Therapeutics, Basilea, Flare Therapeutics, Loxo/LillySpeakers' Bureau: Gilead Sciences, The Lynx GroupResearch Funding: Mirati Therapeutics (Inst), Novartis (Inst), Debiopharm Group (Inst), Bayer (Inst), Janssen (Inst), Seattle Genetics (Inst) Bernard H. BochnerConsulting or Advisory Role: Olympus Arjun V. BalarLeadership: GT BiopharmaStock and Other Ownership Interests: GT BiopharmaHonoraria: Merck, Genentech/Roche, AstraZeneca/MedImmuneConsulting or Advisory Role: Genentech/Roche, Merck, Cerulean Pharma, AstraZeneca/MedImmune, Pfizer/EMD Serono, Incyte, Seattle Genetics/Astellas, Nektar, Dragonfly Therapeutics, GlaxoSmithKline, Bristol Myers Squibb/CelgeneResearch Funding: Merck (Inst), Genentech/Roche (Inst), AstraZeneca/MedImmune (Inst), Seattle Genetics, Gilead Sciences (Inst) Amir MortazaviHonoraria: Motive Medical IntelligenceConsulting or Advisory Role: Seattle Genetics, Debiopharm Group, PfizerResearch Funding: Acerta Pharma (Inst), Genentech/Roche (Inst), Merck (Inst), Novartis (Inst), Seattle Genetics (Inst), Mirati Therapeutics (Inst), Bristol Myers Squibb (Inst), Roche (Inst), Astellas Pharma (Inst), Debiopharm Group (Inst), Debiopharm Group (Inst) Jonathan E. RosenbergHonoraria: UpToDate, Medscape, Peerview, Research To Practice, Intellisphere, Clinical Care Options, Physicans' Education Resource, MJH Life Sciences, EMD SeronoConsulting or Advisory Role: Lilly, Merck, Roche/Genentech, AstraZeneca/MedImmune, Bristol Myers Squibb, Seattle Genetics, Bayer, BioClin Therapeutics, QED Therapeutics, Adicet Bio, Pharmacyclics, western oncolytics, GlaxoSmithKline, Janssen Oncology, Astellas Pharma, Boehringer Ingelheim, Pfizer/EMD Serono, Mirati Therapeutics, Immunomedics, Tyra Biosciences, Infinity PharmaceuticalsResearch Funding: Genentech/Roche (Inst), Seattle Genetics (Inst), Bayer (Inst), AstraZeneca (Inst), QED Therapeutics (Inst), Astellas Pharma (Inst)Patents, Royalties, Other Intellectual Property: Predictor of platinum sensitivity (Inst)No other potential conflicts of interest were reported.

Spermidine and other polyamines alleviate oxidative stress, yet excess spermidine seems toxic to Escherichia coli unless it is neutralized by SpeG, an enzyme for the spermidine N -acetyl transferase function. Thus, wild-type E. coli can tolerate applied exogenous spermidine stress, but Δ speG strain of E. coli fails to do that. Here, using different reactive oxygen species (ROS) probes and performing electron paramagnetic resonance spectroscopy, we provide evidence that although spermidine mitigates oxidative stress by lowering overall ROS levels, excess of it simultaneously triggers the production of superoxide radicals, thereby causing toxicity in the Δ speG strain. Furthermore, performing microarray experiment and other biochemical assays, we show that the spermidine-induced superoxide anions affected redox balance and iron homeostasis. Finally, we demonstrate that while RNA-bound spermidine inhibits iron oxidation, free spermidine interacts and oxidizes the iron to evoke superoxide radicals directly. Therefore, we propose that the spermidine-induced superoxide generation is one of the major causes of spermidine toxicity in E. coli ., Competing Interests: VK, RM, DG, AK, PD, AP, NT, GM, VC, AA, DD No competing interests declared, (© 2022, Kumar et al.)

CD8 + T cell reactivity towards tumor mutation-derived neoantigens is widely believed to facilitate the antitumor immunity induced by immune checkpoint blockade (ICB). Here we show that broadening in the number of neoantigen-reactive CD8 + T cell (NART) populations between pre-treatment to 3-weeks post-treatment distinguishes patients with controlled disease compared to patients with progressive disease in metastatic urothelial carcinoma (mUC) treated with PD-L1-blockade. The longitudinal analysis of peripheral CD8 + T cell recognition of patient-specific neopeptide libraries consisting of DNA barcode-labelled pMHC multimers in a cohort of 24 patients from the clinical trial NCT02108652 also shows that peripheral NARTs derived from patients with disease control are characterised by a PD1 + Ki67 + effector phenotype and increased CD39 levels compared to bystander bulk- and virus-antigen reactive CD8 + T cells. The study provides insights into NART characteristics following ICB and suggests that early-stage NART expansion and activation are associated with response to ICB in patients with mUC., (© 2022. The Author(s).)

An epidemic of mucormycosis followed the second wave of COVID 19 in the state of Uttar Pradesh, India in May 2021. This epidemic, however, had additional challenges to offer in the form of acute shortage of all forms of amphotericin B, posaconazole and isavuconazole. It was, therefore, planned to assess the trends in minimum inhibitory concentration (MIC) of antifungal agents, viz itraconazole and terbinafine, and provide a template for personalized therapy to see whether the results could be translated clinically. This is an observational, single-center study. Samples comprising nasal swab, nasal and paranasal sinus tissue, brain tissue, brain abscess and orbital content, derived from 322 patients from northern India with mucormycosis, of whom 215 were male and 107 were female, were used for analysis. Cultures were identified both by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and conventional methods of identification. Antifungal susceptibility was done for amphotericin B, posaconazole, isavuconazole, itraconazole and terbinafine as per Clinical Laboratory Standard Institute M38-A2. The outcome was identification of the species of mucormycosis and susceptibility to itraconazole and terbinafine besides other primary antifungal agents. Patients or the public were not involved in the design, or conduct, or reporting or in the dissemination plans of our research. Of 322 patients, 203 were culture-positive, of whom 173 were positive by both MALDI-TOF and conventional methods of identification. Final antifungal susceptibility testing was available for 150 patients. The most common Mucorales found to cause this epidemic was Rhizopus oryzae , followed by R. microsporus Amphotericin B, posaconazole and isavuconazole had low MIC values in 98.8% of all Mucorales identified. The MIC of itraconazole was species-dependent. 97.7% of R oryzae had MIC ≤2 µg/mL. However, only 36.5% of R microsporus had MIC ≤2 µg/mL. For terbinafine, 85.2% of R. microsporus had MIC ≤2 µg/mL. We conclude that identification at the species level is required as antifungal susceptibilities seem to be species-dependent. Assessment of the efficacy of itraconazole and terbinafine warrants further studies with clinical assessment and therapeutic drug monitoring as they seem to be potential candidates especially when the primary agents are not available., Competing Interests: Competing interests: None declared., (© American Federation for Medical Research 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

Objective: To evaluate whether urothelial carcinoma (UC) with sarcomatoid differentiation is associated with a lower pathological response rate to neoadjuvant chemotherapy (NAC) and worse oncological outcomes compared to UC without variant histology among patients undergoing radical cystectomy., Patients and Methods: Patients with UC undergoing cystectomy from 1995 to 2018 at the Memorial Sloan Kettering Cancer Centre were identified. Patients with sarcomatoid differentiation at transurethral resection (TUR) or cystectomy, and patients without variant histology were selected. Downstaging from ≥cT2 to ≤pT1N0 defined partial response and pT0N0 defined complete response. Recurrence-free, cancer-specific and overall survival were modelled., Results: We identified 131 patients with sarcomatoid differentiation and 1722 patients without variant histology, of whom 25 with sarcomatoid histology on biopsy and 313 without variant histology received NAC. Those with sarcomatoid differentiation presented with higher consensus tumour stage (94% ≥T2 vs 62%; P < 0.001) and were, therefore, more likely to receive NAC (29% vs 18%; P = 0.003). We found no evidence to support a difference in partial (24% vs 31%) or complete (20% vs 24%) response between patients with sarcomatoid histology and those with pure UC at TUR (P = 0.6). Among patients with sarcomatoid differentiation, 5-year recurrence-free survival was 55% (95% confidence interval [CI] 41-74) among patients receiving NAC and 40% (95% CI 31-52) among patients undergoing cystectomy alone (P = 0.1). Adjusting for stage, nodal involvement, margin status and receipt of NAC, sarcomatoid differentiation was associated with worse recurrence-free (hazard ratio [HR] 1.82, 95% CI 1.39-2.39), disease-specific (HR 1.66, 95% CI 1.23-2.22), and overall survival (HR 1.37, 95% CI 1.06-1.78)., Conclusions: Sarcomatoid differentiation was associated with higher stage at presentation and independently associated with worse survival. Given similar pathological response rates if sarcomatoid differentiation is detected at initial resection, and greater survival among patients receiving NAC, treatment with NAC appears warranted. Other drivers of the poor outcomes of this histology must be investigated., (© 2021 The Authors BJU International © 2021 BJU International.)

DNA-dependent protein kinase (DNA-PK) plays a crucial role in repair of DNA double-strand breaks by facilitating non-homologous end-joining. Inhibitors of DNA-PK have the potential to block DNA repair and enhance DNA-damaging agents. Peposertib (M3814) is a DNA-PK inhibitor that has shown preclinical activity in combination with DNA-damaging agents, including ionizing radiation (IR) and topoisomerase II inhibitors. Here we evaluated the activity of peposertib (M3814) in combination with radiation in a mouse xenograft model of HPV-associated cervical cancer. Athymic nude female mice with established tumors derived from HeLa cells injected into the flank were treated with vehicle alone (n = 3), IR alone (n = 4), and peposertib (M38814) in combination with IR (M3814 + IR; n = 4). While IR alone was associated with a trend towards decreased tumor volume compared with untreated, only the M3814 + IR treatment arm was associated with consistent and significant reduction in tumor burden, which correlated with higher levels of γ-H2AX in tumor cells, a marker of double-strand DNA breaks. Our data support further clinical evaluation of the combination of peposertib (M38814) and IR in cervical cancer., (© 2022. The Author(s).)

c-MYC is deregulated in triple negative breast cancer (TNBC) pointing to be a promising biomarker for breast cancer treatment. Precise level of MYC expression is important in the control of cellular growth and proliferation. Designing of c-MYC -targeted antidotes to restore its basal level of cellular expression holds an optimistic approach towards anti-cancer treatment. MYC transcription is dominantly controlled by Nuclear Hypersensitive Element III-1 (NHE III1 ) upstream of the promoter region possessing G-Quadruplex silencer element ( Pu-27 ). We have investigated the selective binding-interaction profile of a natural phytophenolic compound Curcumin with native MYC G-quadruplex by conducting an array of biophysical experiments and in silico based Molecular Docking and Molecular Dynamic (MDs) simulation studies. Curcumin possesses immense anti-cancerous properties. We have observed significantly increased stability of MYC -G Quadruplex and thermodynamic spontaneity of Curcumin- MYC GQ binding with negative ΔG value. Transcription of MYC is tightly regulated by a complex mechanism involving promoters, enhancers and multiple transcription factors. We have used Curcumin as a model drug to understand the innate mechanism of controlling deregulated MYC back to its basal expression level. We have checked MYC -expression at transcriptional and translational level and proceeded for Chromatin Immuno-Precipitation assay (ChIP) to study the occupancy level of SP1, Heterogeneous nuclear ribonucleoprotein K (hnRNPK), Nucleoside Diphosphate Kinase 2 (NM23-H2) and Nucleolin at NHE III1 upon Curcumin treatment of MDA-MB-231 cells. We have concluded that Curcumin binding tends to drive the equilibrium towards stable G-quadruplex formation repressing MYC back to its threshold-level. On retrospection of the synergistic effect of upregulated c-MYC and BCL-2 in cancer, we have also reported a new pathway [ MYC-E2F-1-BCL-2- axis] through which Curcumin trigger apoptosis in cancer cells.Communicated by Ramaswamy H. Sarma.

Purpose: To investigate clinical and morphologic characteristics of serous retinal disturbances in patients taking extracellular signal-regulated kinase (ERK) inhibitors., Design: Single-center retrospective study of prospectively collected data., Participants: Of 61 patients receiving ERK inhibitors for treatment of metastatic cancer, this study included 40 eyes of 20 patients with evidence of retinopathy confirmed by OCT., Methods: Clinical examination, fundus photography, and OCT were used to evaluate ERK inhibitor retinopathy. The morphologic features, distribution, and location of fluid foci were evaluated serially. Visual acuity (VA) and choroidal thickness measurements were compared at baseline, fluid accumulation, and resolution., Main Outcome Measures: Characteristics of treatment-emergent choroid and retinal OCT abnormalities as compared with baseline OCT findings and the impact of toxicity on VA., Results: Of 20 patients with retinopathy, most showed fluid foci that were bilateral (100%), multifocal in each eye (75%), and with at least 1 focus involving the fovea (95%). All subretinal fluid foci occurred between the interdigitation zone and an intact retinal pigment epithelium. No statistical difference was found in choroidal thickness at fluid accumulation and resolution compared with baseline. Forty-five percent of eyes showed evidence of concomitant intraretinal edema localized to the outer nuclear layer. At the time of fluid accumulation, 57.5% eyes showed a decline in VA (mainly by 1-2 lines from baseline). For all eyes with follow-up, the subretinal fluid and intraretinal edema were reversible and resolved without medical intervention, and best-corrected VA at fluid resolution was not statistically different from baseline. Concomitant intraretinal fluid was not associated with worsening of VA. No patient discontinued or decreased drug dose because of retinopathy., Conclusions: This study showed that ERK inhibitors may cause subretinal fluid foci with unique clinical and morphologic characteristics. The observed foci were similar to mitogen-activated protein kinase kinase (MEK) inhibitor-associated retinopathy and distinct from central serous chorioretinopathy. However, unlike with MEK inhibitors, an increased occurrence of concomitant intraretinal fluid without significant additive visual impact seems to occur with ERK inhibitors. In this series, ERK inhibitors did not cause irreversible loss of vision or serious eye damage; retinopathy was self-limited and did not require medical intervention., (Copyright © 2021 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Importance: Fibroblast growth factor receptor (FGFR) 1 to 4 inhibitors are approved by the US Food and Drug Administration and suppress the mitogen-activated protein kinase (MAPK) pathway, with a potential for treatment-related retinopathy. To date, implications of FGFR inhibitor-associated ocular toxic effects are poorly described. Therefore, more detailed clinical descriptions of this ocular toxic effect could help explain visual symptoms while receiving drug therapy., Objective: To describe the clinical and morphologic characteristics of serous retinal disturbances associated with FGFR inhibitors., Design, Setting, and Participants: In this retrospective case series, 146 patients receiving FGFR inhibitors as cancer treatment at a single tertiary referral center were included. This study included 40 eyes of 20 patients with retinopathy by optical coherence tomography (OCT). OCTs were obtained on the remaining patients and the results were judged normal. Patients were recruited from March 2012 to January 2021., Main Outcomes and Measures: Characteristics of treatment-emergent choroidal and retinal OCT abnormalities as compared with baseline OCT, associated with visual acuity at presentation and at fluid resolution., Results: A total of 20 of 146 patients (13.7%) exhibited FGFR inhibitor-associated retinopathy. Of these 20 patients, 11 (55%) were female, and the median (range) age was 62.6 (42.7-86.0) years. The median (range; mean) time from medication start to initial subretinal fluid detection was 21 (5-125; 32) days. The median (interquartile range [IQR]) baseline logMAR best-corrected visual acuity (BCVA) was 0 (0-0.1). At fluid accumulation, 11 eyes had decreased vision: the median (IQR) subgroup baseline BCVA was 0 (0-0.1); and the median (IQR) BCVA change from baseline to accumulation was -0.1 (-0.2 to -0.1). For 26 eyes (65%) with follow-up, the subretinal fluid resolved without medical intervention or drug interruption in all but 1 patient. At fluid resolution, the median (IQR) BCVA was 0.1 (0-0.1), and the change in median (IQR) BCVA from baseline to fluid resolution was 0 (-0.03 to 0). No patient discontinued drug therapy on account of their retinopathy., Conclusions and Relevance: FGFR inhibitors result in subretinal fluid foci similar to other drugs that inhibit the MAPK pathway. In this series, FGFR inhibitors did not cause irreversible loss of vision; the retinopathy was self-limited and did not require medical intervention. These results may explain visual symptoms while taking the drug, although the precise frequency or magnitude of this adverse effect cannot be determined with certainty from this retrospective investigation.

Eosinophils influence antitumor immunity and may predict response to treatment with immune checkpoint inhibitors (ICIs). To examine the association between blood eosinophil counts and outcomes in patients with advanced or metastatic urothelial carcinoma (mUC) treated with ICIs, we identified 2 ICI-treated cohorts: discovery (n=60) and validation (n=111). Chemotherapy cohorts were used as comparators (first-line platinum-based chemotherapy, n=75; second-line or more pemetrexed, n=77). The primary endpoint was overall survival (OS). Secondary endpoints were time on treatment (ToT) and progression-free survival. Univariate and multivariate analyses were performed using Cox proportional hazard models. Associations between changes in eosinophil count at weeks 2/3 and 6 after the start of ICI treatment were analyzed using landmark analyses. Baseline characteristics of the ICI cohorts were similar. In the discovery cohort, an optimal cutoff for pretreatment eosinophil count was determined [Eos-Lo: <100 cells/µL; n=9 (15%); Eos-Hi: ≥100 cells/µL; n=51 (85%)]. Eos-Lo was associated with inferior outcomes [OS: hazard ratio (HR), 3.98; 95% confidence interval (CI), 1.85-8.56; P<0.013; ToT: HR, 2.45; 95% CI, 1.17-5.10; P=0.017]. This was confirmed in the validation cohort [Eos-Lo: n=17 (15%); Eos-Hi: n=94 (85%)] (OS: HR, 2.51; 95% CI, 1.31-4.80; P=0.006; ToT: HR, 2.22; 95% CI, 1.2-3.80; P=0.004), and remained significant after adjustment for other prognostic factors. Changes in eosinophil counts at weeks 2/3 and 6 were not clearly associated with outcomes. In chemotherapy cohorts, eosinophil counts were not associated with outcomes. In conclusion, low pretreatment eosinophil count was associated with poorer outcomes in patients with mUC treated with ICIs, and may represent a new predictive biomarker., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)