Knockdown of Antisense Noncoding Mitochondrial RNA Reduces Tumorigenicity of Patient-Derived Clear Cell Renal Carcinoma Cells in an Orthotopic Xenograft Mouse Model.

Clear cell renal cell carcinoma (ccRCC) is the most prevalent form of renal cancer and its treatment is hindered by a resistance to targeted therapies, immunotherapies and combinations of both. We have reported that the knockdown of the antisense noncoding mitochondrial RNAs (ASncmtRNAs) with chemically modified antisense oligonucleotides induces proliferative arrest and apoptotic death in tumor cells from many human and mouse cancer types. These studies have been mostly performed in vitro and in vivo on commercially available cancer cell lines and have shown that in mouse models tumor growth is stunted by the treatment. The present work was performed on cells derived from primary and metastatic ccRCC tumors. We established primary cultures from primary and metastatic ccRCC tumors, which were subjected to knockdown of ASncmtRNAs in vitro and in vivo in an orthotopic xenograft model in NOD/SCID mice. We found that these primary ccRCC cells are affected in the same way as tumor cell lines and in the orthotopic model tumor growth was significantly reduced by the treatment. This study on patient-derived ccRCC tumor cells represents a model closer to actual patient ccRCC tumors and shows that knockdown of ASncmtRNAs poses a potential treatment option for these patients.

Caveolin-1-dependent tenascin C inclusion in extracellular vesicles is required to promote breast cancer cell malignancy.

Background: Elevated expression of CAV1 in breast cancer increases tumor progression. Extracellular vesicles (EVs) from CAV1-expressing MDA-MB-231 breast cancer cells contain Tenascin C (TNC), but the relevance of TNC remained to be defined. Methods: EVs were characterized by nanotracking analysis, microscopy and western blotting. The uptake of EVs by cells was studied using flow cytometry. The effects of EVs on breast cancer cells were tested in migration, invasion, colony formation and in vivo assays. Results: EVs were taken up by cells; however, only those containing TNC promoted invasiveness. In vivo, EVs lacking TNC ceased to promote tumor growth. Conclusion: CAV1 and TNC contained in breast cancer cell-derived EVs were identified as proteins that favor progression of breast cancer.

Caveolin-1 (CAV1) is a protein that in breast cancer increases with disease progression. Extracellular vesicles (EVs) from breast cancer cells with CAV1 also contain Tenascin C (TNC) protein, but the importance of TNC remained to be defined. EVs were identified by size, microscopy and protein analysis. The effects of EVs on breast cancer cells were studied using cells and experiments in animals. CAV1 expression promotes TNC inclusion into EVs, which increased the aggressiveness of recipient breast cancer cells. In animals, only EVs with TNC increased features associated with cancer spread, while EVs lacking TNC reduced tumor growth.

EV-miRNA-Mediated Intercellular Communication in the Breast Tumor Microenvironment.

Cancer research has prioritized the study of the tumor microenvironment (TME) as a crucial area of investigation. Understanding the communication between tumor cells and the various cell types within the TME has become a focal point. Bidirectional communication processes between these cells support cellular transformation, as well as the survival, invasion, and metastatic dissemination of tumor cells. Extracellular vesicles are lipid bilayer structures secreted by cells that emerge as important mediators of this cell-to-cell communication. EVs transfer their molecular cargo, including proteins and nucleic acids, and particularly microRNAs, which play critical roles in intercellular communication. Tumor-derived EVs, for example, can promote angiogenesis and enhance endothelial permeability by delivering specific miRNAs. Moreover, adipocytes, a significant component of the breast stroma, exhibit high EV secretory activity, which can then modulate metabolic processes, promoting the growth, proliferation, and migration of tumor cells. Comprehensive studies investigating the involvement of EVs and their miRNA cargo in the TME, as well as their underlying mechanisms driving tumoral capacities, are necessary for a deeper understanding of these complex interactions. Such knowledge holds promise for the development of novel diagnostic and therapeutic strategies in cancer treatment.

Nefrolitotomía Endoscópica ex vivo del injerto renal de donante vivo con instrumento flexible

Introduction: The finding of an asymptomatic stone in the study of a living kidney donor does not necessarily contraindicate donation, however, there is no consensus on the management of these cases. The use of a graft with lithiasis may represent a risk of recurrence in the remaining kidney in the donor and eventual obstructive complications in the transplanted kidney. The objective of this work is to present the usefulness of ureteroscopy (URS) to resolve lithiasis ex vivo before transplantation. Material and Methods: Donors with a small, asymptomatic kidney stone and with an analysis of lithogenic factors without relevant findings were considered to continue in the donation process. The kidney unit with stone was selected for nephrectomy. Results: Four donor kidneys underwent flexible URS after nephrectomy under hypothermic preservation conditions during bench preparation. The average time of the procedure was 35 minutes and the stone was extracted in all cases without incident. The transplant was carried out in the usual way and the evolution of the recipients was without complications and with excellent renal function. During follow-up, no recurrence of lithiasis was observed in donors or recipients. Conclusions: In this experience, the URS of the donor kidney was a feasible procedure and was not associated with adverse consequences for the graft. The main advantage of this procedure is to avoid the potential risk to the recipient of an obstructive graft complication.

[Living-donor kidney graft ex vivo endoscopic nephrolithotomy with flexible instrument]. / Nefrolitotomía Endoscópica ex vivo del injerto renal de donante vivo con instrumento flexible.

INTRODUCTION: The finding of an asymptomatic stone in the study of a living kidney donor does not necessarily contraindicate donation, however, there is no consensus on the management of these cases. The use of a graft with lithiasis may represent a risk of recurrence in the remaining kidney in the donor and eventual obstructive complications in the transplanted kidney. The objective of this work is to present the usefulness of ureteroscopy (URS) to resolve lithiasis ex vivo before transplantation. MATERIAL AND METHODS: Donors with a small, asymptomatic kidney stone and with an analysis of lithogenic factors without relevant findings were considered to continue in the donation process. The kidney unit with stone was selected for nephrectomy. RESULTS: Four donor kidneys underwent flexible URS after nephrectomy under hypothermic preservation conditions during bench preparation. The average time of the procedure was 35 minutes and the stone was extracted in all cases without incident. The transplant was carried out in the usual way and the evolution of the recipients was without complications and with excellent renal function. During follow-up, no recurrence of lithiasis was observed in donors or recipients. CONCLUSIONS: In this experience, the URS of the donor kidney was a feasible procedure and was not associated with adverse consequences for the graft. The main advantage of this procedure is to avoid the potential risk to the recipient of an obstructive graft complication.

Self-Organization Dynamics of Collagen-like Peptides Crosslinking Is Driven by Rose-Bengal-Mediated Electrostatic Bridges.

The present work focuses on the computational study of the structural micro-organization of hydrogels based on collagen-like peptides (CLPs) in complex with Rose Bengal (RB). In previous studies, these hydrogels computationally and experimentally demonstrated that when RB was activated by green light, it could generate forms of stable crosslinked structures capable of regenerating biological tissues such as the skin and cornea. Here, we focus on the structural and atomic interactions of two collagen-like peptides (collagen-like peptide I (CLPI), and collagen-like peptide II, (CLPII)) in the presence and absence of RB, highlighting the acquired three-dimensional organization and going deep into the stabilization effect caused by the dye. Our results suggest that the dye could generate a ternary ground-state complex between collagen-like peptide fibers, specifically with positively charged amino acids (Lys in CLPI and Arg in CLPII), thus stabilizing ordered three-dimensional structures. The discoveries generated in this study provide the structural and atomic bases for the subsequent rational development of new synthetic peptides with improved characteristics for applications in the regeneration of biological tissues during photochemical tissue bonding therapies.

Predicting malignancy in small testicular lesions.

Introduction: Small testicular lesions ≤20 mm (STL) detected by ultrasound (US), usually non-palpable, have been reported to be benign in up to 80% of cases. Thus, partial orchiectomy with or without frozen section examination and surveillance has been advocated for these kinds of lesions. We seek to report the proportion of benign lesions in testicular tumors ≤20 mm detected by US in our population and explore the predicting factors of malignancy. Material and methods: A retrospective descriptive study of orchiectomies performed for testicular tumors in patients older than 15 years between 2005 and 2019 was performed, including all patients with lesions ≤20 mm on US imaging. Results: A total of 70 patients with STL were included (mean age 34.6 ±10.8 years). Overall, 69% of the lesions were malignant while the smallest lesions (≤10 mm) showed 61% of cancer. Moreover, in the subgroup of non-palpable lesions ≤10 mm, 50% were malignant. Multifocal tumors were found in 18 subjects with a malignancy rate of 88%. There was a significant association between maximum size on US, multifocality and malignancy. Neither tumor markers nor palpability foretold a malignant lesion. A predictive model including size and multifocality was created showing a positive predictive value of 83.3%. Conclusions: US maximum size and multifocality were predictors of malignancy in STL. However, even the smallest lesions showed a 50% chance of being malignant, thus surgery with or without intraoperative biopsy is warranted in most cases.

Smartphone screen testing, a novel pre-diagnostic method to identify SARS-CoV-2 infectious individuals.

The COVID-19 pandemic will likely take years to control globally, and constant epidemic surveillance will be required to limit the spread of SARS-CoV-2, especially considering the emergence of new variants that could hamper the effect of vaccination efforts. We developed a simple and robust - Phone Screen Testing (PoST) - method to detect SARS-CoV-2-positive individuals by RT-PCR testing of smartphone screen swab samples. We show that 81.3-100% of individuals with high-viral-load SARS-CoV-2 nasopharyngeal-positive samples also test positive for PoST, suggesting this method is effective in identifying COVID-19 contagious individuals. Furthermore, we successfully identified polymorphisms associated with SARS-CoV-2 Alpha, Beta, and Gamma variants, in SARS-CoV-2-positive PoST samples. Overall, we report that PoST is a new non-invasive, cost-effective, and easy-to-implement smartphone-based smart alternative for SARS-CoV-2 testing, which could help to contain COVID-19 outbreaks and identification of variants of concern in the years to come.

Protective role of intravesical BCG in COVID-19 severity.

OBJECTIVES: To establish the role of BCG instillations in the incidence and mortality of COVID-19. PATIENTS AND METHODS: NMIBC patients in instillations with BCG (induction or maintenance) during 2019/2020 were included, establishing a COVID-19 group (with a diagnosis according to the national registry) and a control group (NO-COVID). The cumulative incidence (cases/total patients) and the case fatality rate (deaths/cases) were established, and compared with the national statistics for the same age group. T-test was used for continuous variables and Fisher's exact test for categorical variables. RESULTS: 175 patients were included. Eleven patients presented CIS (11/175, 6.3%), 84/175 (48.0%) Ta and 68/175 (38.9%) T1. Average number of instillations = 13.25 ± 7.4. One hundred sixty-seven patients (95.4%) had complete induction. Forty-three patients (cumulative incidence 24.6%) were diagnosed with COVID-19. There is no difference between COVID-19 and NO-COVID group in age, gender or proportion of maintenance completed. COVID-19 group fatality rate = 1/43 (2.3%). Accumulated Chilean incidence 70-79 years = 6.3%. Chilean fatality rate 70-79 years = 14%. CONCLUSIONS: According to our results, patients with NMIBC submitted to instillations with BCG have a lower case-fatality rate than the national registry of patients between 70 and 79 years (2.3% vs. 14%, respectively). Intravesical BCG could decrease the mortality due to COVID-19, so instillation schemes should not be suspended in a pandemic.

Intrarenal temperature measurement associated with holmium laser intracorporeal lithotripsy in an ex vivo model.

INTRODUCTION: The aim of this article was to quantify the effect of the use of holmium laser during intracorporeal lithotripsy in an ex vivo model. MATERIAL AND METHODS: A simulated model for laser nephro-lithotripsy was designed. Two ex vivo porcine kidneys were used. Electronic thermometer electrodes were inserted on the upper calyx. Intracorporeal lithotripsy was simulated with a holmium laser. Intrarenal temperature was recorded both at the beginning and after one minute of laser use with delta temperature (DT) defined as the difference between them. Measurements were made at different irrigation heights (30, 40, and 50 cm H2O), frequency (Hz), and laser energy (J) in addition to the presence or absence of the access sheath. Analysis of factors associated with temperature change was performed. RESULTS: Thirty-eight observations were recorded. The measurement without the use of access sheath showed an average DT of 4.9, 5.1, and 6.5°C for 5, 10, and 15 Hz, respectively; however, with a sheath, DTs were 0.2, 0.5, and 1.5°C. In terms of energy, mean DTs of 4.3, 6.1, 5.2, and 13.9°C for 0.5, 0.8, 1.0, and 1.5 J were recorded; in contrast, with a sheath, averages of 0.4, 0.5, 0.5, and 3.8°C, respectively were noted. In the adjusted model, energy, frequency, and use of sheath and water height were significant. CONCLUSIONS: The configuration of the laser significantly modifies the intrarenal temperature and height of the bladder irrigation. The use of an access sheath provides lower intrarenal temperatures regardless of laser configuration and water height.