The ancestral ESCRT protein TOM1L2 selects ubiquitinated cargoes for retrieval from cilia.

Many G protein-coupled receptors (GPCRs) reside within cilia of mammalian cells and must undergo regulated exit from cilia for the appropriate transduction of signals such as hedgehog morphogens. Lysine 63-linked ubiquitin (UbK63) chains mark GPCRs for regulated removal from cilia, but the molecular basis of UbK63 recognition inside cilia remains elusive. Here, we show that the BBSome-the trafficking complex in charge of retrieving GPCRs from cilia-engages the ancestral endosomal sorting factor target of Myb1-like 2 (TOM1L2) to recognize UbK63 chains within cilia of human and mouse cells. TOM1L2 directly binds to UbK63 chains and the BBSome, and targeted disruption of the TOM1L2/BBSome interaction results in the accumulation of TOM1L2, ubiquitin, and the GPCRs SSTR3, Smoothened, and GPR161 inside cilia. Furthermore, the single-cell alga Chlamydomonas also requires its TOM1L2 ortholog in order to clear ubiquitinated proteins from cilia. We conclude that TOM1L2 broadly enables the retrieval of UbK63-tagged proteins by the ciliary trafficking machinery.

A Red-Emitting Fluorescence Sensor for Detecting Boronic Acid-Containing Agents in Cells.

The amount and localization of boron-10 atoms delivered into tumor cells determines the therapeutic effect of boron neutron capture therapy (BNCT) and, consequently, efforts have been directed to develop fluorescence sensors to detect intracellular boronic acid compounds. Currently, these sensors are blue-emitting and hence are impracticable for co-staining with nucleus staining reagents, such as DAPI and Hoechst 33342. Here, we designed and synthesized a novel fluorescence boron sensor, BS-631, that emits fluorescence with a maximum emission wavelength of 631 nm after reaction with the clinically available boronic acid agent, 4-borono-l-phenylalanine (BPA). BS-631 quantitatively detected BPA with sufficiently high sensitivity (detection limit = 19.6 µM) for evaluating BNCT agents. Furthermore, BS-631 did not emit fluorescence after incubation with metal cations. Notably, red-emitting BS-631 could easily and clearly visualize the localization of BPA within cells with nuclei co-stained using Hoechst 33342. This study highlights the promising properties of BS-631 as a versatile boron sensor for evaluating and analyzing boronic acid agents in cancer therapy.

Perception of risk factors for cancer in the ABC population.

OBJECTIVE To evaluate the knowledge about risk factors for cancer in patients treated at the ABC Medical School (FMABC). METHODS Cross-sectional observational study conducted in 2019. The American Cancer Institute's Cancer Risk Awareness Survey questionnaire was used with 29 cancer risk factors, 14 of which were proven to cause cancer and 15 without consensus or scientific evidence of causality with cancer but that are often reminded by most of the population. Qualitative variables were described by frequency and percentage, and quantitative variables by mean and standard deviation or median and range depending on normality, assessed by the Shapiro-Wilk test. The study was conducted in accordance with the Helsinki Declaration for Research and approved by the Research Ethics Committee. RESULTS 191 patients were included. Median age 54 (20 to 90), 64% female. 35.6% reported current or previous smoking. 3.1% consumed alcohol more than 5 drinks/week. 56% reported sedentary lifestyle. 44% had at least 1 case of cancer in relatives up to 2nd degree. The average of correct answers in the analyzed population was 12.83 ± 3.06. A weak positive correlation was observed between income and number of cases (rho = 0.177, p = 0.02). No relationship was observed between the number of correct answers and level of education, age, sex, marital status, race or patients with a positive family history for cancer. CONCLUSION The knowledge about risk factors for cancer in the ABC population is low, which may contribute to the adoption of risk behaviors for the disease.

Perception of risk factors for cancer in the ABC population

SUMMARY OBJECTIVE To evaluate the knowledge about risk factors for cancer in patients treated at the ABC Medical School (FMABC). METHODS Cross-sectional observational study conducted in 2019. The American Cancer Institute's Cancer Risk Awareness Survey questionnaire was used with 29 cancer risk factors, 14 of which were proven to cause cancer and 15 without consensus or scientific evidence of causality with cancer but that are often reminded by most of the population. Qualitative variables were described by frequency and percentage, and quantitative variables by mean and standard deviation or median and range depending on normality, assessed by the Shapiro-Wilk test. The study was conducted in accordance with the Helsinki Declaration for Research and approved by the Research Ethics Committee. RESULTS 191 patients were included. Median age 54 (20 to 90), 64% female. 35.6% reported current or previous smoking. 3.1% consumed alcohol more than 5 drinks/week. 56% reported sedentary lifestyle. 44% had at least 1 case of cancer in relatives up to 2nd degree. The average of correct answers in the analyzed population was 12.83 ± 3.06. A weak positive correlation was observed between income and number of cases (rho = 0.177, p = 0.02). No relationship was observed between the number of correct answers and level of education, age, sex, marital status, race or patients with a positive family history for cancer. CONCLUSION The knowledge about risk factors for cancer in the ABC population is low, which may contribute to the adoption of risk behaviors for the disease.

RESUMO OBJETIVO Avaliar o conhecimento sobre fatores de risco para câncer em pacientes atendidos nos ambulatórios da Faculdade de Medicina do ABC (FMABC). MÉTODOS Estudo transversal e observacional conduzido em 2019. Foi utilizado o questionário Cancer Risk Awarness Survey do American Institute for Cancer Research com 29 fatores de risco para câncer, sendo 14 fatores comprovadamente causadores de câncer e 15 sem consenso ou evidência científica de causalidade com o câncer, mas que são frequentemente citados pela população. Variáveis qualitativas foram descritas por frequência e porcentagem, e variáveis quantitativas por média e desvio padrão ou mediana e intervalo a depender da normalidade, avaliada pelo teste de Shapiro-Wilk. Estudo realizado de acordo com a Declaração de Helsinque para pesquisa e aprovado pelo Comitê de Ética em Pesquisa. RESULTADOS Foram incluídos 191 pacientes. Mediana de idade 54 (20 a 90), 64% do sexo feminino; 35,6% reportaram tabagismo atual ou anterior; 3,1% consumiam mais que cinco doses de álcool por semana; 56% reportaram sedentarismo; 44% tinham pelo menos um caso de câncer em parentes de até segundo grau. A média de acertos na população analisada foi 12,83 ± 3,06. Foi observada correlação positiva fraca entre renda e número de acertos (rho=0,177, p=0,02). Não foram observadas relações entre o número de acertos e nível de escolaridade, idade, sexo, estado civil, cor ou pacientes com história familiar positiva para câncer. CONCLUSÃO O conhecimento sobre fatores de risco para câncer na população do ABC é baixo, o que pode contribuir para a adoção de comportamentos de risco para a doença.

Establishment of a new technique for the fabrication of regenerative cartilage with a microslicer device to prepare three dimensional diced cartilage.

Chondrocytes are utilized to cartilage regeneration by being harvested through enzymatic digestion and expanded by monolayer culture. However, these procedures will cause deterioration and dedifferentiation of the chondrocytes. In addition, scaffolds are often needed to provide the cartilage with mechanical strength and three-dimensional structures. We tried to use diced cartilage prepared using a micro-slicer without digestion, monolayer culture or scaffolds. In this study, an appropriate culture condition to induce the fusion of diced cartilage in vitro and cartilage regeneration in vitro and in vivo was determined to realize a scaffold-free cartilage regeneration. As a result, diced cartilages aggregated when they were cultured more than 5 weeks in the media containing 10% fetal bovine serum (FBS). Diced cartilage cultured for 7 weeks with the media containing 10%, followed by the culture with the media containing insulin-like growth factor-1 for 5 weeks in the ultralow attachment plate showed most prominent cartilage formation both in vitro and in vivo. The volume of regenerated cartilage was 2.14 times larger than that of the original cartilage. These results indicated that large regenerative cartilage from a small amount of cartilage was achieved without deterioration or dedifferentiation.