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Background: Non-muscle invasive bladder cancer (NMIBC) patients are affected by a high risk of recurrence. The topography of collagen fibers represents a hallmark of the neoplastic extracellular microenvironment. Objective: Assess the topographic change associated with different stages of bladder cancer (from neoplastic lesions to bona fide tumor) and whether those changes favour the development of NMIBC. Design, Setting, and Participants: Seventy-one clinical samples of urothelial carcinoma at different stages were used. Topographic changes preceding tumor onset and progression were evaluated in the rat bladder cancer model induced by nitrosamine (BBN), a bladder-specific carcinogen. The preclinical model of actinic cystitis was also used in combination with BBN. Validated hematoxylin-eosin sections were used to assess the topography of collagen fibrils associated with pre-tumoral steps, NMIBC, and MIBC. Findings: Linearization of collagen fibers was higher in Cis and Ta vs. dysplastic urothelium, further increased in T1 and greatest in T2 tumors. In the BBN preclinical model, an increase in the linearization of collagen fibers was established since the beginning of inflammation, such as the onset of atypia of a non-univocal nature and dysplasia, and further increased in the presence of the tumor. Linearization of collagen fibers in the model of actinic cystitis was associated with earlier onset of BBN-induced tumor. Conclusions: The topographic modification of the extracellular microenvironment occurs during the inflammatory processes preceding and favoring the onset of bladder cancer. The topographic reconfiguration of the stroma could represent a marker for identifying and treating the non-neoplastic tissue susceptible to tumor recurrence.

Background: Fabry disease (FD) is a multisystem, monogenic, X-linked storage disorder caused by mutations in the GLA gene, resulting in reduced alfa-galactosidase A enzyme activity. This effect leads to the accumulation of glycosphingolipids, particularly globotriosylceramide, in various tissues, including the heart, kidney, vasculature, smooth muscle and peripheral nervous system. Hemizygous males are usually more severely affected than females, in whom random inactivation of an X-chromosome may lead to variable phenotype. Summary: Among the manifestations of FD, exercise intolerance is commonly diagnosed but often underestimated, even though it significantly limits quality of life, especially in young patients. This review primarily discusses the various pathophysiological mechanisms involved in exercise intolerance in FD patients, such as altered muscle composition, compromised cardiopulmonary framework, and peripheral neuropathy. Secondarily, it explores the potential effect of available therapy, including Enzyme Replacement Therapy (ERT) and chaperone therapy (migalastat), in reducing exercise intolerance while considering the potential impact of physical activity and exercise training as adjunctive treatments. Conclusion: Exercise intolerance has a major impact on the well-being of people with Fabry disease. Exercise training can play an important role in addition to drug therapy.

Abstract Bladder mechanical properties are critical for organ function and tissue homeostasis. Therefore, alterations of tissue mechanics are linked to disease onset and progression. This study aims to characterize the tissue elasticity of the murine bladder wall considering its different anatomical components, both in healthy conditions and in actinic cystitis, a state characterized by tissue fibrosis. Here, we exploit Brillouin microscopy, an emerging technique in the mechanobiology field that allows mapping tissue mechanics at the microscale, in non-contact mode and free of labeling. We show that Brillouin imaging of bladder tissues is able to recognize the different anatomical components of the bladder wall, confirmed by histopathological analysis, showing different tissue mechanical properties of the physiological bladder, as well as a significant alteration in the presence of tissue fibrosis. Our results point out the potential use of Brillouin imaging on clinically relevant samples as a complementary technique to histopathological analysis, deciphering complex mechanical alteration of each tissue layer of an organ that strongly relies on mechanical properties to perform its function.

Abstract In the complex tumor microenvironment (TME), mesenchymal cells are key players, yet their specific roles in prostate cancer (PCa) progression remain to be fully deciphered. This study employs single-cell RNA sequencing to delineate molecular changes in tumor stroma that influence PCa progression and metastasis. Analyzing mesenchymal cells from four genetically engineered mouse models (GEMMs) and correlating these findings with human tumors, we identify eight stromal cell populations with distinct transcriptional identities consistent across both species. Notably, stromal signatures in advanced mouse disease reflect those in human bone metastases, highlighting periostin’s role in invasion and differentiation. From these insights, we derive a gene signature that predicts metastatic progression in localized disease beyond traditional Gleason scores. Our results illuminate the critical influence of stromal dynamics on PCa progression, suggesting new prognostic tools and therapeutic targets.

Paratuberculosis (Johne’s disease) is a globally widespread infectious disease affecting domestic and wild ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). The bacterium is excreted in the feces and is characterized by high environmental resistance. The new Animal Health Law (Regulation EU 2016/429) on transmissible animal diseases, recently in force throughout the European Union, includes paratuberculosis within the diseases requiring surveillance in the EU, listing some domestic and wild Bovidae, Cervidae, and Camelidae as potential reservoirs. Taking advantage of a culling activity conducted in the Stelvio National Park (Italy), this study investigated MAP infection status of red deer (Cervus elaphus) between 2018 and 2022, and evaluated the probability of being MAP-positive with respect to individual and sampling-level variables. A total of 390 subjects were examined macroscopically and tested for MAP, using different diagnostic tools: IS900 qPCR, culture, histopathology, and serology. Twenty-three of them were found positive for MAP by at least one test, with an overall prevalence of 5.9% (95% CI 4.0–8.7), that, respectively, ranged from 12.4% in the first culling season to 2.0 and 2.1% in the 2019–2020 and 2021–2022 culling seasons. Quantitative PCR assay on ileocecal valve and mesenteric lymph nodes detected the highest number of MAP positive animals. The results of the study showed the increased probability of being MAP-positive with increasing age and that red deer with lower body mass values were more likely to be infected with MAP. Overall, the absence of signs of clinical paratuberculosis and gross lesions together with the low level of shedding witness early phases of the disease among the positive red deer and support an improvement of the paratuberculosis status of this population, as shown by the decreased prevalence of the disease over the years.

Introduction The clinical benefit of the anti-CTLA-4 monoclonal antibody (mAb) ipilimumab has been well established but limited by immune-related adverse events, especially when ipilimumab is used in combination with anti-PD-(L)1 mAb therapy. To overcome these limitations, we have developed XTX101, a tumor-activated, Fc-enhanced anti-CTLA-4 mAb.Methods XTX101 consists of an anti-human CTLA-4 mAb covalently linked to masking peptides that block the complementarity-determining regions, thereby minimizing the mAb binding to CTLA-4. The masking peptides are designed to be released by proteases that are typically dysregulated within the tumor microenvironment (TME), resulting in activation of XTX101 intratumorally. Mutations within the Fc region of XTX101 were included to enhance affinity for FcγRIII, which is expected to enhance potency through antibody-dependent cellular cytotoxicity.Results Biophysical, biochemical, and cell-based assays demonstrate that the function of XTX101 depends on proteolytic activation. In human CTLA-4 transgenic mice, XTX101 monotherapy demonstrated significant tumor growth inhibition (TGI) including complete responses, increased intratumoral CD8+T cells, and regulatory T cell depletion within the TME while maintaining minimal pharmacodynamic effects in the periphery. XTX101 in combination with anti-PD-1 mAb treatment resulted in significant TGI and was well tolerated in mice. XTX101 was activated in primary human tumors across a range of tumor types including melanoma, renal cell carcinoma, colon cancer and lung cancer in an ex vivo assay system.Conclusions These data demonstrate that XTX101 retains the full potency of an Fc-enhanced CTLA-4 antagonist within the TME while minimizing the activity in non-tumor tissue, supporting the further evaluation of XTX101 in clinical studies.

Aim: Hematopoietic Stem Cell Transplantation (HSCT) is used to treat or cure a wide spectrum of conditions. Cryopreservation maintains the therapeutic properties of Peripheral Blood Stem Cells (PBSC), allowing clinical, regulatory, and logistical procedures needed for successful HSCT. Complications of mobilizing and collecting additional PBSC following recovery from the myeloablative HSCT have led centers to adopt the practice of collection and storage of enough PBSC for two HSCTs. However, the collection, cryopreservation, and storage of extra PBSC than that needed for a single HSCT have logistical, infrastructure, and cost issues. Here, we evaluated the collection, storage, and utilization practices of PBSC for two or more HSCTs. Methods: This cross-sectional study included patients referred for HSCT at eight transplant centers. Cryopreservation, storage, and other laboratory procedures were conducted at Cetebio/Fundação Hemominas between 2013 and 2022. Frozen cells were stored in the vapor phase of nitrogen. The records were reviewed to obtain information the regarding the patients and frozen cell therapy products. Results: Cetebio released frozen PBSC for 1,311 transplantations between 2013 and 2022. Two hundred fifty patients had 321 bags with residual PBSC in storage after their first HSCT (234 autologous and 16 allogeneic HSCT). Patients had a mean age of 49.6 ± 3.4 years, and most were male (162; 64.8%). Most (200; 80%) had Multiple Myeloma (MM), followed by leukemia (18; 7.2%), Germ Cell Tumor (GCT, 14; 5.6%), and other diagnoses (18; 7.2%). Cells from 66 (26.4%), 163 (65.2%), and 21 (8.4%) patients were frozen 2 years or less ago, 2.1 to 7 years ago, and more than 7 years ago, respectively. Thirty-three bags from 24 (9.6%) patients were released for a second HSCT within a median of 2.2 years after the cryopreservation. The mean age of patients at the time of the second HSCT (23 autologous and one allogenic transplantation) was 51.6 ± 14.4 years, ranging from 18 to 69 years. Most were male (15; 62.5%) and had MM (15; 62.5%) followed by GCT (5; 20.8%), leukemia (3; 12,5%), and myelodysplastic syndrome (1; 4.2%). The remaining 288 frozen bags continue stored at our facility and represent about a third of our tank storage capacity. The human, infrastructure, and economic resources required to cryopreserve and store unused PBSC are equivalent to the resources needed to attend nearly 144 extra patients. Discussion: The frequency of second transplantation was similar to previously published studies (9% to 19%). This study is limited by its retrospective design and the relatively short-term follow-up. Some frozen PBSCs will probably be released for a second HSCT in the future. Nevertheless, in public healthcare systems, resources should be used based on equity and effectiveness. Since most patients probably never undergo a second HSCT, our institution changed its practices regarding PBSC collection, cryopreservation, and storage to support the second HSCT. Currently, it is allowed only for diseases that benefit from tandem infusions. Conclusions: this study showed a low utilization of stored PBSC to support the second HSCT. The results suggest a review of practices regarding cryopreservation PBSCs for a second HSCT, especially in public healthcare system settings and places where novel agents and new therapeutic approaches for treatment of MM are available.

Introduction: Knowledge of cryopreservation of Hematopoietic Progenitor Cells (HPC) collected by Bone Marrow aspiration (HPC,BM) is limited. Aim: To describe the experience of a Cell Processing Center with HPC,BM cryopreservation for therapeutic use, and to compare the BMT outcomes with non-cryopreserved historical controls. Methods: This is a nested case-control study. The casuistry consisted of 16 HPC,BM units cryopreserved between 2020 and 2022. The HPC,BM units were Red Blood Cells (RBC) depleted using Hydroxyethyl Starch (HES) 450/0.7, followed by centrifugation for volume reduction. The units were frozen in a mechanical freezer at -80°C after adding the cryopreservation solution (5%HES/5%DMSO) and stored in the LN2 vapor phase. Data from patients whose HPC,BM unit was RBC depleted and freshly infused were used as historical controls. The control for one case of cryopreserved HPC,BM for autologous transplantation was one cryopreserved HPC,PB. Controls were matched to cases by diagnosis. Results: The mean age of cases was 29.4 ± 11.1 years. Nine (56.3%) patients were female. One (6.3%) unit was cryopreserved for autologous use and fifteen (93.8%) for unrelated allogeneic BMT. One (6.3%) patient had neuroblastoma, one (6.3%) SCID, one (6.3%) CML, three (18.8%) AML, one (6.3%) Griscelli syndrome, two (12.5%) Wiskott-Aldrich syndrome and four (25%) ALL. Twelve (75%) units were released for therapeutic use, of which ten (83.3%) already have grafting data available. The mean time to neutrophil and platelet engraftment was 16.8 and 18.4 days, respectively. One (11.1%) patient had graft failure. There was no difference in time to neutrophil and platelet engraftment between cases and controls. Related to adverse effects during infusion, the frequency of nausea and hemoglobinuria was higher in controls (40% vs. 14.3%, respectively), while the frequency of hypertension was higher in cases (20% vs. 57.1%, respectively), although not significant. The occurrence of fever was significantly higher in controls when compared to cases (60% vs. 0%, p = 0.045). Although not significant, pseudomembranous colitis (0% vs. 25%, respectively) was more frequent in cases, while mucositis (50% vs. 12.5%, respectively) was more frequent in controls. Acute GVHD was not observed in cases and controls during the observational period. Cox regression analysis adjusted for age, sex, transplant center, and infused CD34+ cell dose showed no significant difference in the occurrence of death or grafting failure between cases and controls. Discussion: The higher frequency of fever in the controls can be explained by the fresh infusion of the product collected in an open system with frequently observed contamination. Cryopreservation allows clinical, regulatory, and logistical procedures, including cell therapy product quality control testing. Therefore, cryopreservation permits the identification and sensitivity of the microorganism in case of a positive test and the knowledge of results by the transplant staff. There was no difference in the other transplant outcomes when cases were compared with controls. However, the low number of individuals may have resulted in a lack of statistical power to detect associations, especially for variables with missing observations. Conclusion: This study indicates that HPC,BM cryopreservation was not associated with an increased occurrence of adverse effects during infusion, BMT complications, and graft failure or death.

Background and Objectives. This exploratory study aimed to evaluate the potential usefulness of channeled information for ten questions from scientists. The study’s objectives were to 1) assess the correspondence of channeled and non-channeled answers within questions, 2) evaluate the correspondence of different channelers’ responses for each question while in channeling and non-channeling states, 3) examine whether channelers believe they are receiving information from the same source, and 4) explore qualitative themes that emerge for each question. Method. Fifteen channelers provided answers to 10 questions in a channeled and non-channeled state. The first three objectives were quantitatively evaluated by three judges using structured criteria to assess correspondence. The last objective employed qualitative thematic analysis of the channeled answers. Results. The quantitative analyses found 1) low correspondence between channeled and non-channeled answers as hypothesized, 2) virtually no correspondence for each question across channelers, contrary to our hypothesis, and 3) little support that the channelers perceived they were accessing the same source of information. The qualitative analysis resulted in coherent and common themes in the channeled responses for many but not all ten questions. Several methodological lessons were gleaned from the study, such as refining inclusion/exclusion criteria and the questions asked of channelers, allowing a similar amount of time for channeled and non-channeled answers, ensuring consistent methods across study sites, and including additional quantitative measures informing on the channelers experience in channeling and non-channeling states. Conclusions. This exploratory study offers insight into improving future studies attempting to obtain valuable information through channeling.

Foamy viruses are ancient retroviruses that are prevalent in non-human primates. Fernández et al. report an X-ray structure of a protein domain from a simian Foamy virus that mediates the attachment to cells, providing insights into viral entry.

The micromechanical fingerprints of bladders in rats change following actinic cystitis or in the presence of bladder tumours.

Abstract Background Metastatic castration-resistant prostate cancer remains a challenging condition to treat. Among the available therapeutic options, the androgen receptor signaling inhibitors abiraterone acetate plus prednisone (AA) and enzalutamide (Enza), are currently the most used first-line therapies in clinical practice. However, validated clinical indicators of prognosis in this setting are still lacking. In this study, we aimed to evaluate a prognostic model based on the time of metastatic disease presentation (after prior local therapy [PLT] or de-novo [DN]) and disease burden (low volume [LV] or high-volume [HV]) at AA/Enza onset for mCRPC patients receiving either AA or Enza as first-line. Methods A cohort of consecutive patients who started AA or Enza as first-line treatment for mCRPC between January 1st, 2015, and April 1st, 2019 was identified from the clinical and electronic registries of the 9 American and European participating centers. Patients were classified into 4 cohorts by the time of metastatic disease presentation (PLT or DN) and volume of disease (LV or HV; per the E3805 trial, HV was defined as the presence of visceral metastases and/or at least 4 bone metastases of which at least 1 out the axial/pelvic skeleton) at AA/Enza onset. The endpoint was overall survival defined as the time from AA or Enza initiation, respectively, to death from any cause or censored at the last follow-up visit, whichever occurred first. Results Of the 417 eligible patients identified, 157 (37.6%) had LV/PLT, 87 (20.9%) LV/DN, 64 (15.3%) HV/PLT, and 109 (26.1%) HV/DN. LV cohorts showed improved median overall survival (59.0 months; 95% CI, 51.0–66.9 months) vs. HV cohorts (27.5 months; 95% CI, 22.8–32.2 months; P = 0.0001), regardless of the time of metastatic presentation. In multivariate analysis, HV cohorts were confirmed associated with worse prognosis compared to those with LV (HV/PLT, HR = 1.87; p = 0.029; HV/DN, HR = 2.19; P = 0.002). Conclusion Our analysis suggests that the volume of disease could be a prognostic factor for patients starting AA or Enza as first-line treatment for metastatic castration-resistant prostate cancer, pending prospective clinical trial validation.

In this report, we look at the relationship between prior circumcision and presenting stage of penile cancer. We performed an analysis of an international, multicenter database of 1254 penile cancer patients diagnosed from 1980 to 2019 in the United States, Europe, Brazil, and China, and analyzed the relationship between circumcision and presenting T and N stage. A total of 710 patients met the inclusion criteria and were statistically analyzed. We found that uncircumcised men with locally advanced tumors (T3–T4) had significantly higher risk of lymph node metastasis compared with circumcised men.

Article Highlights This paper aims at presenting a comprehensive LCA case study that uses real-world operations data to investigate the environmental impacts of the High V.LO-City hydrogen fuel cell bus system (i.e. H2FC bus, integrating hydrogen production and delivery options and refuelling stations operation) against a conventional Euro-6 Diesel bus. The High V.LO-City project consisted of the integrated testing of new generation hydrogen buses and their hydrogen production and refuelling infrastructure in some European cities. It differs from other demonstration projects because it intends to evaluate both the operation of the vehicles and the production and supply of hydrogen to use them as substitutes for conventional diesel buses and trolleybuses. Fuel economy during bus operation is around 10.25 KgH2/100 km, and the refuelling station energy demand ranges between 7 and 9 KWh/KgH2. The results clearly indicate that it is advantageous to promote such mode of transportation in areas where electricity is primarily produced from renewables (i.e. wind power as an alternative to fossil fuels). However, potential setbacks, in particular those related to problem shifting and to grid improvement, should be addressed accordingly.

Detection and removal of bladder cancer lesions at an early stage is crucial for preventing tumor relapse and progression. This study aimed to develop a new technological platform for the visualization of small and flat urothelial lesions of high-grade bladder carcinoma in situ (CIS).We found that the integrin α5β1, overexpressed in bladder cancer cell lines, murine orthotopic bladder cancer and human bladder CIS, can be exploited as a receptor for targeted delivery of GNRs functionalized with the cyclic CphgisoDGRG peptide (Iso4). The GNRs@Chit-Iso4 was stable in urine and selectively recognized α5β1 positive neoplastic urothelium, while low frequency ultrasound-assisted shaking of intravesically instilled GNRs@Chit-Iso4 allowed the distribution of nanoparticles across the entire volume of the bladder. Photoacoustic imaging of GNRs@Chit-Iso4 bound to tumor cells allowed for the detection of neoplastic lesions smaller than 0.5 mm that were undetectable by ultrasound imaging and bioluminescence.

Nanoparticles (NPs) are paving the way for improved treatments for difficult to treat diseases diseases; however, much is unknown about their fate in the body. One important factor is the interaction between NPs and blood proteins leading to the formation known as the “protein corona” (PC). The PC, consisting of the Hard (HC) and Soft Corona (SC), varies greatly based on the NP composition, size, and surface properties. This highlights the need for specific studies to differentiate the PC formation for each individual NP system. This work focused on comparing the HC and SC of three NPs with different matrix compositions: a) polymeric NPs based on poly(lactic-co-glycolic) acid (PLGA), b) hybrid NPs consisting of PLGA and Cholesterol, and c) lipidic NPs made only of Cholesterol. NPs were formulated and characterized for their physico-chemical characteristics and composition, and then were incubated in human plasma. In-depth purification, identification, and statistical analysis were then performed to identify the HC and SC components. Finally, similar investigations demonstrated whether the presence of a targeting ligand on the NP surface would affect the PC makeup. These results highlighted the different PC fingerprints of these NPs, which will be critical to better understand the biological influences of the PC and improve future NP designs.

Objectives: Lymph node invasion (LNI) is related to long-term survival in patients with muscle-invasive bladder cancer. However, in the case of variant histology (VH), data on pelvic lymph node dissection (PLND) and LNI are sparse. We described the pattern of care of PLND in patients with VHs of bladder cancer, exploring predictors of LNI. Methods: Using the 2001–2016 SEER registry, 20 767 bladder cancer patients who underwent PLND were identified. Included histological variants were pure urothelial carcinoma (UC), micropapillary UC, sarcomatoid UC, lymphoepithelioma-like UC, adenocarcinoma, sarcoma, giant and spindle cell carcinoma, squamous cell carcinoma (SCC), and neuroendocrine tumor. Uni- and multivariable logistic regression analyses tested for LNI predictors. Cox regression was used to test for predictors of overall mortality (OM) among both LNI positive and LNI negative patients. Results: Overall, 2464 (11.9%) harbored a VH. On multivariate analysis, only micropapillary UC was associated with higher risk (OR = 3.39) of LNI. This association was maintained when only the subset of patients treated without perioperative chemotherapy were analyzed (OR = 3.30). Similarly, higher T stage (T2 stage OR = 2.24; T3–4 stage OR = 9.44) and the use of chemotherapy (OR = 2.29) were associated with a higher risk of LNI. Among patients with LNI (5299, 25.5%), SCC (HR = 1.87), T3–4 stage (HR = 1.94), age at diagnosis (HR = 1.01) and geographic region (south) (HR = 1.22) were predictors of higher risk of OM. Conversely, chemotherapy (HR = 0.69) and number of removed LN (HR = −0.99) were associated with lower risk of OM. Finally, in a subgroup of patients without LNI, sarcomatoid UC (HR = 1.58) and giant and spindle cell carcinoma (HR = 1.83) were the only VH predictors of OM. Conclusions: We described different patterns of care in patients with VHs of bladder cancer. Micropapillary UC was an independent risk factor for LNI. Among patients harboring LNI, those with SCC VH had higher OM compared to pure UC. Conversely, sarcomatoid UC and giant and spindle cell carcinoma were predictors of OM in patients without nodal involvement.

Molecular mechanisms associated with human germ cell aplasia in infertile men remain undefined. Here the authors perform single-cell transcriptome profiling to highlight differentially expressed genes and pathways in each somatic cell type in testes of men with idiopathic germ cell aplasia.

In this Review, Laura Martinez-Vidal et al. summarize current knowledge of factors affecting extracellular matrix (ECM) remodeling in the context of pathological mechanical changes to tissue properties. They focus on the importance of research on tissue stiffness to the development of diagnostic tools and therapies to treat urological disease.

BackgroundAdult primary retroperitoneal sarcomas (RPSs) are a group of heterogeneous tumors with different histological subtypes. Comprehensive genomic profiling (CGP) analyses have recently provided significant insights into the biology of sarcomas by identifying genomic alterations (GAs) which could benefit from targeted therapies. MethodsRPS were evaluated by CGP using next-generation sequencing of up to 406 cancer-related genes. Tumor mutational burden (TMB) was determined on 0.83 to 1.14 mut/Mb of sequenced DNA. Finally, PD-L1 expression was determined. ResultsOverall, 296 cases of primary RPS were analyzed. Liposarcoma (LPS) subtype had more GA/tumor than leiomyosarcoma (LMS) subtypes, with follicular dendritic cell sarcomas harboring the highest and synovial sarcomas the lowest. TP53 and Rb1 alterations were the highest in LMS, and CDK4/6 and MDM2 in LPS. However, both the TMB and targetable GA rates were low across subtypes. PD-L1 immunostaining was low positive in 21% and high positive in 5% of patients, respectively. ConclusionsCGP analysis revealed that potentially actionable genomic targets were rare in our cohort of RPS. Moreover, RPSs seem less likely to respond to immune checkpoint inhibitors based on putative biomarkers status. Nevertheless, genomic stratification according to histological subtypes led to description of GAs that can inform future clinical trials design.

We present the case of a 45-year-old Caucasian woman who attended the Endocrinology Unit for a left cervical mass discovered during follow-up for autoimmune chronic thyroiditis. The ultrasound-guided fine-needle aspiration biopsy of the lesion was consistent with a metastasis of follicular thyroid carcinoma. The sonographic neck evaluation revealed no thyroid nodules but three markedly hypoechoic and highly vascularized areas, with irregular margins and hyperechoic spots. In the clinical suspicion of primary thyroid neoplasm, ultrasound-guided fine-needle aspiration biopsy of two of the three areas was performed, but both cytological reports were non-diagnostic, revealing only colloid and blood. Subsequently, the patient underwent surgical removal of the cervical mass, with the intra-operatory consultation with frozen section examination suggesting follicular-like neoplasia. For this reason, thyroidectomy with both central and lateral neck dissection was performed. Surprisingly, the final histologic examination revealed chronic thyroiditis in the thyroid specimen and no evidence of metastasis in the left neck mass. Consequently, the pathological revision of the frozen section assessment led to the final diagnosis of chronic thyroiditis on the lateral ectopic thyroid. This case represents an uncommon example of lateral ectopic thyroid tissue with coexisting normally located thyroid tissue both affected by chronic thyroiditis.

Lakis et al. report novel findings related to the epigenetic landscape of pancreatic neuroendocrine cancer. This relatively large cohort provides functional insights into the epigenetic wiring of pancreatic neuroendocrine tumor sub-types with the potential ability to stratify tumours of different prognosis.

We present the case of a 69-year-old woman who attended the Endocrinology Unit of Modena for a suspicious lymph node in the left cervical compartment discovered during the follow-up of a recurrent gynecological malignancy. At neck ultrasonography, a thyroid goiter was detected, and the further cytological examination was inconclusive for thyroid nodule and compatible with a localization of an adenocarcinoma with papillary architecture for the lymph node. The histological examination after a left neck dissection confirmed the presence of an intracapsular metastasis of a papillary carcinoma immunohistochemically focally positive for thyroid transcription factor 1 and paired box 8 and negative for thyroglobulin. Subsequently, in the suspicion of a thyroid primitiveness, a total thyroidectomy was performed, revealing an intraparenchymal follicular variant of papillary thyroid carcinoma of 2 mm in the right lobe. During the follow-up, the appearance of a suspected cervical metastatic lesion led to another neck dissection, histologically compatible with a papillary carcinoma localization, immunohistochemically focally positive for thyroid transcription factor 1 and paired box 8, and negative for thyroglobulin. The histological revision of surgical specimens suggests the cervical recurrence of the prior gynecological cancer, rather than a thyroid carcinoma metastasis. The case described shows how carefully the cytological, histological and immunoistochemical results must be evaluated in oncological management, considering the whole patient’s history.

Testicular volume (TV) is considered a good clinical marker of hormonal and spermatogenic function. Accurate reference values for TV measures in infertile and fertile men are lacking. We aimed to assess references values for TV in white-European infertile men and fertile controls. We analyzed clinical and laboratory data from 1940 (95.0%) infertile men and 102 (5.0%) fertile controls. Groups were matched by age using propensity score weighting. TV was assessed using a Prader orchidometer (PO). Circulating hormones and semen parameters were investigated in every male. Descriptive statistics, Spearman's correlation, and logistic regression models tested potential associations between PO-estimated TV values and clinical variables. Receiver operating characteristic (ROC) curves were used to find TV value cutoffs for oligoasthenoteratozoospermia (OAT) and nonobstructive azoospermia (NOA) status in infertile men. The median testicular volume was smaller in infertile than that of fertile men (15.0 ml vs 22.5 ml; P < 0.001). TV positively correlated with total testosterone, sperm concentration, and progressive sperm motility (all P ≤ 0.001) in infertile men. At multivariable logistic regression analysis, infertile status (P < 0.001) and the presence of left varicocele (P < 0.001) were associated with TV < 15 ml. Testicular volume thresholds of 15 ml and 12 ml had a good predictive ability for detecting OAT and NOA status, respectively. In conclusion, infertile men have smaller testicular volume than fertile controls. TV positively correlated with total testosterone, sperm concentration, and progressive motility in infertile men, which was not the case in the age-matched fertile counterparts.

Abstract Background Few clinical trials have investigated the prevention of radiographic progression in children with juvenile idiopathic arthritis treated with antirheumatic drugs. This study aimed to investigate radiographic progression in patients with systemic juvenile idiopathic arthritis (sJIA) and patients with polyarticular-course juvenile idiopathic arthritis (pcJIA) treated with the anti–interleukin-6 receptor antibody tocilizumab for 2 years in the TENDER and CHERISH randomized controlled trials, respectively. Methods Standard radiographs of both wrists and both hands in the posteroanterior view were obtained within 4 weeks of baseline and were repeated at weeks 52 ± 4 and 104 ± 4 in both trials. All films were scored by two independent readers using the adapted Sharp–van der Heijde (aSH) and Poznanski scoring methods. Although the Poznanski score indicates bone growth limitation or cartilage growth decrease, which are not the same as joint space narrowing in rheumatoid arthritis, its change reflects damage to cartilage. Therefore, impairment in the Poznanski score as well as the aSH score was considered as a measure of structural joint damage. Radiographic progression was defined as worsening of radiographic scores beyond the smallest detectable difference. Results Poznanski and aSH scores were available at baseline and at one or more postbaseline time points for 33 and 47 of 112 sJIA patients and 61 and 87 of 188 pcJIA patients, respectively, providing a representative subset of the study populations. The inter-reader and intra-reader agreement intra-class correlation coefficient was > 0.8. Median baseline Poznanski and aSH scores, respectively, were − 2.4 and 24.6 for sJIA patients and − 1.5 and 8.0 for pcJIA patients. Compared with baseline, aSH scores remained stable for all sJIA patients at week 52, whereas 9.4% of sJIA patients had radiographic progression according to Poznanski scores at week 52; at 104 weeks, radiographic progression according to aSH and Poznanski scores was observed in 5.4% and 11.5%, respectively. In pcJIA patients, radiographic progression from baseline at 52 weeks and at 104 weeks was 12.5% and 2.9%, respectively, using aSH scoring and 6.5% and 4%, respectively, using Poznanski scoring. Conclusion Tocilizumab may delay radiographic progression in children with sJIA and children with pcJIA. Trial registration Trial registration numbers and dates: TENDER, NCT00642460 (March 19, 2008); CHERISH, NCT00988221 (October 1, 2009)

Introdução: O transplante de medula óssea (TMO) autólogo é uma modalidade de tratamento para vários tipos de doença. A predição do sucesso na mobilização pode ser útil para otimizar a coleta de células progenitoras hematopoéticas (CPH). O objetivo foi identificar características que podem influenciar a eficácia da mobilização e criar um modelo de predição de sucesso da mobilização em candidatos ao TMO do estado de Minas Gerais. Materiais e métodos: Tratou-se de estudo retrospectivo, sendo que foram analisados dados dos pacientes candidatos ao TMO autólogo entre 09/2015 e 12/2019. Os participantes foram assistidos em cinco hospitais e os procedimentos laboratoriais foram realizados no Centro de Processamento Celular (CPC) do Cetebio/Fundação Hemominas. Os prontuários dos participantes foram revistos para obtenção das seguintes informações: sexo, idade, peso, diagnóstico, esquema de mobilização, ciclo de mobilização, número de esquemas prévios de quimioterapia, radioterapia prévia e sorologia positiva. Os dados laboratoriais analisados foram contagem de células CD34+ e o hemograma da amostra de sangue periférico (SP) pré-coleta. A contagem de células CD34+ foi realizada usando o protocolo ISHAGE. O hemograma foi realizado utilizando contador automatizado (SYSMEX XN-1000 AS-01). O desfecho estudado, mobilização boa, foi definido como a contagem de células CD34+ viáveis ≥ 20/uL no SP. A regressão logística foi usada para determinar o efeito independente de cada covariável no desfecho estudado. Resultados: a população do estudo consistiu em 559 pacientes, sendo 297 (53,1%) do sexo masculino. A mediana de idade foi 54 (20) anos. O mieloma múltiplo foi o diagnóstico mais comum (n = 323; 57,8%), seguido de linfoma não Hodgkin (n = 106; 19%) e linfoma de Hodgkin (n = 100; 17,9%). Dos 559 participantes, 321 (57,4%) eram bons mobilizadores. Das variáveis laboratoriais, a viabilidade das células CD34+ (OR = 1,2; IC95%: 1,11-1,30; p < 0,001), o VCM (OR = 0,92; IC95%:0,87-0,95; p < 0,001), o nRBC (OR = 3006,5; IC95%: 42,1-214.711,3; p < 0,001) e a contagem de granulócitos imaturos (OR = 1,43; IC95%: 1,30-1,57; p < 0,001) foram significativamente associadas à mobilização boa no modelo multivariado. Além disso, os participantes com diagnóstico de mieloma múltiplo apresentaram chance 2,8 vezes (OR = 2,82; IC95%: 1,76-4,5; p < 0,001) maior de serem bons mobilizadores se comparado aos pacientes com linfoma. Participantes que realizaram a coleta no primeiro ciclo de mobilização apresentaram chance 3,6 vezes (OR = 3,63; IC95%: 1,59-8,28; p = 0,002) maior de serem bons mobilizadores. Discussão: O CPC do Cetebio atende nove centros transplantadores. Os centros que realizam a coleta das CPH estão localizados entre 40 a 568 km de distância do Cetebio (local de quantificação de células CD34+). A predição do desfecho da mobilização pode permitir a adoção de medidas complementares como, por exemplo, o uso de plerixafor. Além disso, permitiria otimizar a coleta de CPH por aférese. Assim, seria possível evitar dias adicionais de coleta, reduzindo custos e riscos relacionados à administração de doses adicionais de G-CSF, manutenção de cateter e internação hospitalar prolongada. Conclusão: O sucesso na mobilização foi maior nos participantes que fizeram o primeiro ciclo de mobilização e que apresentavam diagnóstico de mieloma múltiplo. Além disso, níveis maiores de nRBC, granulócitos imaturos e viabilidade das células CD34+, bem como níveis menores de VCM foram associados ao sucesso na mobilização.

Introdução: Devido à pandemia do novo Coronavírus, a criopreservação tem sido recomendada para garantir a continuidade do transplante de medula óssea (TMO) alogênico não aparentado independentemente da fonte de células. Porém, a criopreservação de células progenitoras hematopoéticas coletadas por punção aspirativa da medula óssea (CPH,MO) é menos utilizada e poucos dados estão disponíveis na literatura. Objetivo: descrever a experiência do Centro de Processamento Celular do Cetebio/Fundação Hemominas com a criopreservação de CPH,MO para utilização terapêutica em TMO. Métodos: Trata-se de um estudo do tipo série de casos e a casuística foi composta por oito unidades de CPH,MO criopreservadas entre 10/2020 e 06/2021. As unidades de CPH,MO foram submetidas à deseritrocitação utilizando hidroxietilamido (HES) 450/0,7 e, em seguida, à centrifugação para desplasmatização. O volume da camada leucocitária foi ajustado para atingir a concentração de células nucleadas final ≤2 × 108 NC/mL após a adição da solução de criopreservação (5%HES/5%DMSO). As unidades foram congeladas em freezer mecânico a -80°C e armazenadas em tanque a vapor de nitrogênio. Resultados: oito unidades de CPH,MO foram criopreservadas para utilização em nove pacientes; uma unidade foi dividida para uso em dois pacientes pediátricos irmãos HLA idênticos. Uma (11,1%) unidade foi criopreservada para o uso autólogo e oito (88,9%) para uso alogênico não aparentado. Cinco (55,6%) pacientes eram do sexo masculino. A média de idade dos pacientes foi 14,7 ± 19,1 anos (1–42). Um (11,1%) paciente possuía neuroblastoma, um (11,1%) LMC, um (11,1%) LMA, um (11,1%) síndrome de Griscelli, dois (22,2%) síndrome de Wiskott-Aldrich e três (33,3%) LLA. O volume (mL) médio das unidades pré e pós-processamento foi 756,9 ± 411,7 e 78,7 ± 45,8, respectivamente. O volume (mL) médio de hemácias pré e pós-processamento foi 239,1 ± 150,8 e 27,02 ± 16,9, respectivamente. A média do total de células nucleadas (x108) pré e pós-processamento foi 153,07 ± 80,0 e 120,01 ± 67,7, respectivamente. A média de células CD34+ viáveis (x106) pré e pós-processamento foi 131,7 ± 74,0 e 120,2 ± 77,8, respectivamente. Quatro (50%) unidades apresentaram teste microbiológico positivo (Staphylococcus hominis, Staphylococcus epidermidis, Corynebacterium sp e Staphylococcus aureus). A viabilidade celular (%) média pós-descongelamento em amostras de segmento foi 66,1 ± 8,5. Os ensaios clonogênicos realizados em amostras pré-processamento (n = 6), pós-processamento (n = 8) e pós-descongelamento (n = 7) apresentaram crescimento positivo. Seis (66,6%) unidades foram liberadas para uso terapêutico, das quais cinco (83,3%) possuímos os dados da enxertia. Nenhum paciente apresentou falha de enxertia ou óbito após a infusão. O tempo médio de enxertia de granulócitos e plaquetas foi 17 ± 5,8 e 19,5 ± 9,2 dias, respectivamente. Discussão: A redução do volume possibilitou a criopreservação do material, sendo que a recuperação celular foi satisfatória. Os testes de controle de qualidade realizados em todas etapas do processo evidenciaram a qualidade e segurança do procedimento e foram corroborados pelos dados de enxertia. Alertamos, entretanto, sobre a necessidade de cuidados adicionais com os pacientes devido ao volume de hemácias a ser infundido e à contaminação bacteriana, mais frequentemente observada nas unidades de CPH,MO. Conclusões: A criopreservação CPH, MO apresentou resultados satisfatórios, demonstrando que a metodologia utilizada é segura e eficiente.