Targeting the serrated pathway of colorectal cancer with mutation in BRAF

A recently acknowledged morphological pathway to colorectal cancer originates from precursor polyps with a serrated appearance due to branching and folding of the colon epithelium. This serrated origin accounts for up to 30% of all colorectal tumors but these are heterogeneous regarding molecular characteristics and patient outcome. Here we review the current knowledge about the classification of this tumor subtype and its association with five key features: mutation status of the BRAF or KRAS genes, the CpG island methylation phenotype, microsatellite instability, immune cell infiltration, and overexpression of GTPase RAC1b. Subsequently, available therapeutic approaches for targeting these molecular characteristics are presented and critically discussed.

The relationship between health literacy and indicators of informed decision making for colorectal cancer screening among African Americans

African Americans are disproportionately affected by colorectal cancer (CRC) incidence and mortality. CRC early detection leads to better treatment outcomes and, depending on the screening test, can prevent the development of CRC. African Americans, however, are screened less often than Whites. Aspects of decision making (e.g., decisional conflict, decision self-efficacy) can impact decision making outcomes and may be influenced by social determinants of health, including health literacy. However the relationship between social determinants of health and indicators of decision making in this population is not fully understood. Additionally, individuals have a choice between different CRC screening tests and an individual’s desire to use a particular screening test may be associated with social determinants of health such as health literacy. This study aimed to examine the relationship between social determinants of health and indicators of decision making for CRC screening among African Americans. A total of 111 participants completed a baseline and 14-month follow-up survey assessing decisional conflict, decision self-efficacy, decisional preference (shared versus informed decision making), and CRC test preference. Health literacy was negatively associated with decisional conflict and positively associated with decision self-efficacy (ps < .05). Individuals who were unemployed or working part-time had significantly greater decisional conflict than individuals working full-time (ps < .05). Individuals with a first-degree family history of CRC had significantly lower decision self-efficacy than individuals without a family history (p < .05). Women were significantly more likely to prefer making a shared decision rather than an informed decision compared to men (p < .05). Lastly, previous CRC screening behavior was significantly associated with CRC test preference (e.g., individuals previously screened using colonoscopy were significantly more likely to prefer colonoscopy for their next screening test; ps < .05). These findings begin to identify social determinants of health (e.g., health literacy, employment) that are related to indicators of decision making for CRC among African Americans. Furthermore, these findings suggest further research is needed to better understand these relationships to help with the future development and improvement of interventions targeting decision making outcomes for CRC screening in this population.

The third dimension: new developments in cell culture models for colorectal research

Cellular models are important tools in various research areas related to colorectal biology and associated diseases. Herein, we review the most widely used cell lines and the different techniques to grow them, either as cell monolayer, polarized two-dimensional epithelia on membrane filters, or as three-dimensional spheres in scaffoldfree or matrix-supported culture conditions. Moreover, recent developments, such as gut-on-chip devices or the ex vivo growth of biopsy-derived organoids, are also discussed. We provide an overview on the potential applications but also on the limitations for each of these techniques, while evaluating their contribution to provide more reliable cellular models for research, diagnostic testing, or pharmacological validation related to colon physiology and pathophysiology.

Tumor-related splicing variant RAC1b in colorectal cancer: regulation and role in tumorigenesis

Purpose: Alternative splicing of the small GTPase RAC1 generates RAC1b, a hyperactivated variant that is overexpressed in a subtype of colorectal tumors. The objective of our studies is to understand the molecular regulation of this alternative splicing event and how it contributes to tumorigenesis. Experimental description: The regulation of the RAC1b splicing event in human colon cell lines was dissected using a transfected RAC1 minigene and the role of upstream regulating protein kinases through an RNA interference approach. The functional properties of the RAC1b protein were characterized by experimental modulation of Rac1b levels in colon cell lines. Results: The RAC1b protein results from an in-frame inclusion of an additional alternative exon encoding 19 amino acids that change the regulation and signaling properties of the protein. RAC1b is a hyperactive variant that exists predominantly in the GTP-bound active conformation in vivo and promotes cell cycle progression and cell survival through activation of the transcription factor NF-κB. RAC1b overexpression functionally cooperates with the oncogenic mutation in BRAF-V600E to sustain colorectal tumor cell survival. The splicing factor SRSF1 was identified to bind an exonic splice enhancer element in the alternative exon and acts as a prime regulator of Rac1b alternative splicing in colorectal cells. SRSF1 is controlled by upstream protein kinase SRPK1, the inhibition or depletion of which led to reduced SRSF1 phosphorylation and nuclear translocation with a concomitant reduction in RAC1b levels. As further SRSF1-regulating pathways we discovered kinase GSK3 and a cyclooxygenase independent effect of the non-steroidal anti-inflammatory drug ibuprofen. Conclusions: Expression of tumor-related RAC1b in colorectal cancer depends critically on SRSF1 for the observed deregulation of alternative splicing during tumorigenesis and is controlled by upstream protein kinases that can be pharmacologically targeted.

Ibuprofen Inhibits Overexpression of Tumor-Related Rac1b through SRSF1

Abstract: The serrated pathway to colorectal tumor formation involves oncogenic mutations in the BRAF gene, which are sufficient for initiation of hyperplastic growth but not for tumor progression. A previous analysis of colorectal tumors revealed that overexpression of splice variant Rac1b occurs in around 80% of tumors with mutant BRAF and both events proved to cooperate in tumor cell survival. Patients with inflamed human colonic mucosa also have increased expression of Rac1b as well as mice with experimentally induced colitis. The increase of Rac1b in the mouse model was specifically prevented by the nonsteroidal anti-inflammatory drug ibuprofen. Purpose: The objective of our study is to understand the molecular regulation of Rac1b alternative splicing event and how it contributes to tumorigenesis. Experimental description: HT29 colorectal cell line was used as model to test several signaling pathways after 48h of treatment with ibuprofen. For this we analyzed the proteins of interest by Western Blot and the transcript levels by RT-PCR. Results: Mechanistic studies in cultured HT29 colorectal tumor cells revealed that ibuprofen inhibited Rac1b expression in a cyclooxygenase inhibition–independent manner and targets directly the alternative splicing event. Here, we provide evidence that ibuprofen leads to a decrease in expression of SRSF1, a splicing factor that we previously identified to promote Rac1b alternative splicing. Together, our results suggest that stromal cues, namely, inflammation, can trigger changes in Rac1b expression in the colon and identify ibuprofen as a highly specific and efficient inhibitor of Rac1b overexpression in colorectal tumors. Conclusions: Our data identify an additional cyclooxygenase–independent action of ibuprofen and suggest it may be beneficial in the treatment of patients with the subtype of BRAF-mutated serrated colorectal tumors.

Expression of tumor-related Rac1b antagonizes B-Raf-induced senescence in colorectal cells

Mutations in the BRAF oncogene have been identified as a tumor-initiating genetic event in mainly melanoma, thyroid and colon cancer, resulting in an initial proliferative stimulus that is followed by a growth arrest period known as oncogene-induced senescence (OIS). It remains unknown what triggers subsequent escape from OIS to allow further tumor progression. A previous analysis revealed that overexpression of splice variant Rac1b occurs in around 80% of colorectal tumors carrying a mutation in BRAF. Using both BRaf-V600E-directed RNAi and overexpression we demonstrate that this mutation does not directly lead to Rac1b overexpression, indicating the latter as an independent event during tumor progression. Nonetheless, we observed that expression of oncogenic BRaf-V600E in non-transformed colonocytes (NCM460 cell line) increased both the transcript and protein levels of p14ARF, p15INK4b and p21CIP1 and led to increased expression of β-galactosidase, all indicators of OIS induction. Interestingly, whereas the protein levels of these markers were reduced upon Rac1b overexpression, the levels of their respective transcripts remained unchanged. Importantly, the co-expression of Rac1b with B-Raf-V600E reverted the OIS phenotype, reducing the expression levels of the cell-cycle inhibitors and β-galactosidase to those of control cells. These data identify increased Rac1b expression as one potential mechanism by which colorectal tumor cells can escape from B-Raf-induced OIS.

Nuclear alterations in nasal mucosa epithelial cells of students exposed to formaldehyde

Introduction: Formaldehyde is a compound with a wide range and is commonly used in anatomy and pathology laboratories. At room temperature is quickly volatilized to a pungent and suffocating gas and its inhalation has been correlated to nuclear alterations in different tissues. We aimed to investigate whether exposure to this compound was correlated with the appearance of cytotoxic and genotoxic features in the nasal epithelial cells of students enrolled in a human anatomy course. Material and Methods: This prospective study collected periodically nasal cells from mucosa of 17 volunteers from two different undergraduate programs with different workloads of practical lessons in an anatomy laboratory, 30 and 90 hours per semester. Cells were staining according to Feulgen method and nuclear morphology was analyzed to detect possible damage. Dunn's post hoc test was used in the statistical analysis. Pearson's correlation was performed for gender, age and questionnaire responses. Results: Epithelial cells showed indicators of cytotoxicity and mutagenicity. Students with a more extensive workload in anatomy laboratory displayed a more severe profile with an increase in karyorrhexis (p < 0.05) over time. The micronucleus analysis showed difference between first and second collection (p < 0.01), although it was not maintained over the time. Students with a less extensive workload display no differences in most of cytological features. Despite karyorrhexis was present in a greater number of cells, for this group no significant difference was observed between any range. The same was observed to karyolysis and micronucleus (p > 0.05). Conclusion: Individuals exposed for short periods of time to formaldehyde are subject to the toxic action of this gas. Karyorrhexis was the most frequently observed cytotoxic feature and micronucleus showed an increase between the first time point. The patterns observed between the student's groups suggest a negative effect due to exposure time.

Ibuprofen inhibits overexpression of tumor-related Rac1b through SRSF1

Introduction: The serrated pathway to colorectal tumor formation involves oncogenic mutations in the BRAF gene, which are sufficient for initiation of hyperplastic growth but not for tumor progression. A previous analysis of colorectal tumors revealed that overexpression of splice variant Rac1b occurs in around 80% of tumors with mutant BRAF and both events proved to cooperate in tumor cell survival. Patients with inflamed human colonic mucosa also have increased expression of Rac1b as well as mice with experimentally induced colitis. The increase of Rac1b in the mouse model was specifically prevented by the nonsteroidal anti-inflammatory drug ibuprofen.

Olfactomedin-4 is a candidate biomarker of solid gastric, colorectal, pancreatic, head and neck, and prostate cancers

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Composição racial e a avaliação da reação intradérmica de Montenegro em portadores da leishmaniose cutâneo-mucosa

Em um estudo retrospectivo, indivíduos com a forma mucosa da leishmaniose foram pareados (sexo, idade e ocupação) a outros pacientes com a forma cutânea em atividade, não demonstrando-se diferença significante da freqüência nos grupos raciais. Do mesmo modo, o teste intradérmico de Montenegro não se mostrou diferente nos diversos grupos raciais, entre os 96 pares de pacientes estudados. Uma maior enduração foi observada em pacientes com o acometimento mucoso quando comparado aos cutâneos. _______________________________________________________________________________________ ABSTRACT

Enhanced flat adenoma detection rate with high definition colonoscopy plus i-scan for average-risk colorectal cancer screening

Background and aim: The usefulness of high definition colonoscopy plus i-scan (HD+i-SCAN) for average-risk colorectal cancer screening has not been fully assessed. The detection rate of adenomas and other measurements such as the number of adenomas per colonoscopy and the flat adenoma detection rate have been recognized as markers of colonoscopy quality. The aim of the present study was to compare the diagnostic performance of an HD+i-SCAN with that of standard resolution white-light colonoscope. Methods: This is a retrospective analysis of a prospectively collected screening colonoscopy database. A comparative analysis of the diagnostic yield of an HD+i-SCAN or standard resolution colonoscopy for average-risk colorectal screening was conducted. Results: During the period of study, 155/163 (95.1%) patients met the inclusion criteria. The mean age was 56.9 years. Sixty of 155 (39%) colonoscopies were performed using a HD+i-SCAN. Adenoma-detection-rates during the withdrawal of the standard resolution versus HD+i-SCAN colonoscopies were 29.5% and 30% (p = n.s.). Adenoma/colonoscopy values for standard resolution versus HD+i-SCAN colonoscopies were 0.46 (SD = 0.9) and 0.72 (SD = 1.3) (p = n.s.). A greater number of flat adenomas were detected in the HD+i-SCAN group (6/60 vs. 2/95) (p < .05). Likewise, serrated adenomas/polyps per colonoscopy were also higher in the HD+i-SCAN group. Conclusions: A HD+i-SCAN colonoscopy increases the flat adenoma detection rate and serrated adenomas/polyps per colonoscopy compared to a standard colonoscopy in average-risk screening population. HD+i-SCAN is a simple, available procedure that can be helpful, even for experienced providers. The performance of HD+i-SCAN and substantial prevalence of flat lesions in our average-risk screening cohort support its usefulness in improving the efficacy of screening colonoscopies.

Pérdida de tolerancia inmune en la etiología de las úlceras aftosas recidivantes (RAU) de la mucosa oral: la ruptura de la tolerancia inmunológica causaría injuria persistente en la mucosa bucal provocando las úlceras aftosas recurrentes (2ª parte)

Se ha realizado una revisión de teorías de pérdida de tolerancia inmune publicadas tratando de encontrar aquellos conceptos modernos. Las mismas se refieren a la predisposición genética, influencia de la epigenética en la modelación de un fenotipo vulnerable, las hipótesis de higiene y de microbiota, síndrome de sensibilidad química múltiple, stress crónico, cortisol, el eje hipófisis, hipotálamo y páncreas, óxido nítrico, ataque a la membrana celular, e injuria por reperfusión. Aplicando los principios básicos de las teorías consultadas se demuestra la pérdida de homeostasis, general o parcial, que podría llegar a explicar tres características fundamentales de las úlceras recurrentes orales: dolor, vulnerabilidad y recurrencia.

Surgical site infection in colorectal surgery

Surgical Site Infection is one of the most common infection associated with health care, but can also be the most preventable situation. Surgical site infection in patients undergoing colorectal surgery varies according to the literature, from 3.5 to 21.3%, being identiied as the highest rate of infection among elective procedures and emergency. Objectives: To identify and characterize the occurrence of surgical site infection in patients undergoing colorectal surgery at a hospital in northern Portugal. Methods: A prospective study in a hospital in the north of Portugal in 2015, patients admitted to the surgical service who underwent colorectal surgery. Patients were selected more than 24 hours of admission, obtaining a sample of 102 participants. The characterization of the patient and the surgery was done using a search in the irst 24 hours after surgery and the registration of the infection at the time of occurrence and 30 days after the intervention. Results: 102 participants, 67 (65.7%) were male with a mean age of 71.92 years (37-93 years) and 40.2% underwent emergency surgery. There was a prevalence of surgical site infection in 21 patients (20.6%). Among these 15 were male (71.4%) with mean age of 72.24 years. They were hospitalized on average 22 days, with an average of 19 days of hospitalization after surgery. Escherichia-coli was the microorganism most frequently isolated in culture studies with 13 (60.0%) cases of surgical site infection and organ/space was the main site identiied with infection - 38.1%. Conclusions: The prevalence of surgical site infection was 5.1% and Escherichia coli most common etiologic agent. It is suggested that other studies can analyze the associated factors with this type of infection.