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Nonsense-mediated RNA decay (NMD) is recognized as an RNA surveillance pathway that targets aberrant mRNAs with premature translation termination codons (PTC) for degradation, however, its molecular mechanisms and roles in health and disease remain incompletely understood. In this study, we developed a novel reporter system to accurately measure NMD activity in individual cells. A genome-wide CRISPR-Cas9 knockout screen using this reporter system identified novel NMD-promoting factors, including multiple components of the SF3B complex and other U2 spliceosome factors. Interestingly, cells with mutations in the spliceosome genes SF3B1 and U2AF1, which are commonly found in myelodysplastic syndrome (MDS) and cancers, have overall attenuated NMD activity. Compared with wild-type (WT) cells, SF3B1- and U2AF1-mutant cells were more sensitive to NMD inhibition, a phenotype that is accompanied by elevated DNA replication obstruction, DNA damage, and chromosomal instability. Remarkably, the sensitivity of spliceosome mutant cells to NMD inhibition was rescued by overexpression of RNase H1, which removes R-loops in the genome. Together, these findings shed new light on the functional interplay between NMD and RNA splicing and suggest a novel synthetic lethal strategy for the treatment of MDS and cancers with spliceosome mutations.Significance:This study has developed a novel NMD reporter system and identified a potential therapeutic approach of targeting the NMD pathway to treat cancer with spliceosome gene mutations.

Nonsense-mediated RNA decay (NMD) is recognized as an RNA surveillance pathway that targets aberrant mRNAs with premature translation termination codons (PTC) for degradation, however, its molecular mechanisms and roles in health and disease remain incompletely understood. In this study, we developed a novel reporter system to accurately measure NMD activity in individual cells. A genome-wide CRISPR-Cas9 knockout screen using this reporter system identified novel NMD-promoting factors, including multiple components of the SF3B complex and other U2 spliceosome factors. Interestingly, cells with mutations in the spliceosome genes SF3B1 and U2AF1, which are commonly found in myelodysplastic syndrome (MDS) and cancers, have overall attenuated NMD activity. Compared with wild-type (WT) cells, SF3B1- and U2AF1-mutant cells were more sensitive to NMD inhibition, a phenotype that is accompanied by elevated DNA replication obstruction, DNA damage, and chromosomal instability. Remarkably, the sensitivity of spliceosome mutant cells to NMD inhibition was rescued by overexpression of RNase H1, which removes R-loops in the genome. Together, these findings shed new light on the functional interplay between NMD and RNA splicing and suggest a novel synthetic lethal strategy for the treatment of MDS and cancers with spliceosome mutations. Significance: This study has developed a novel NMD reporter system and identified a potential therapeutic approach of targeting the NMD pathway to treat cancer with spliceosome gene mutations.

The aim of this study is to determine the demographic, pathological, and treatment-related factors that predict recurrence and survival in a Trinidadian cohort of breast cancer patients. The inclusion criteria for this study were female, over 18 years, and with a primary breast cancer diagnosis confirmed by a biopsy report occurring between 2010 and 2015 at Sangre Grande Hospital, Trinidad. Univariate associations with 5-year recurrence-free survival and 5-year overall survival were calculated using the Kaplan–Meier method for categorical variables and Cox Proportional Hazards for continuous variables. A multivariate model for prediction of recurrence and survival was determined using Cox regression. For the period 2010–2015, 202 records were abstracted. Five-year overall survival and recurrence-free survival rates were found to be 74.3% and 56.4%, respectively. Median times from first suspicious finding to date of biopsy report, date of surgery, and date of chemotherapy were 63 days, 125 days, and 189 days, respectively. In the univariate analysis, age (p = 0.038), stage (p

Nonsense-mediated RNA decay (NMD) is well recognized as an RNA surveillance pathway that targets aberrant mRNAs with premature translation termination codons (PTCs) for degradation; however, its molecular mechanisms and roles in health and disease remain incompletely understood. In this study, we developed a novel reporter system that can accurately measure NMD activity in individual cells. By carrying out a genome-wide CRISPR/Cas9 knockout screen using this reporter system, we identified novel NMD-promoting factors, including multiple components of the SF3B complex and other U2 spliceosome factors. Interestingly, we also found that cells with mutations in the U2 spliceosome genes SF3B1 and U2AF1—which are commonly found in myelodysplastic syndrome (MDS) and cancers—have overall attenuated NMD activity. Furthermore, we found that compared to wild type cells, SF3B1 and U2AF1 mutant cells are more sensitive to NMD inhibition, a phenotype that is accompanied by elevated DNA replication obstruction, DNA damage and chromosomal instability. Remarkably, the sensitivity of spliceosome mutant cells to NMD inhibition could be rescued by overexpression of RNase H1, which removes R-loops in the genome. Together, our findings shed new light on the functional interplay between NMD and RNA splicing and suggest a novel strategy for the treatment of MDS and cancers with spliceosome mutations.

Nonsense-mediated RNA decay (NMD) is recognized as an RNA surveillance pathway that targets aberrant mRNAs with premature translation termination codons (PTC) for degradation; however, its molecular mechanisms and roles in health and disease remain incompletely understood. In this study, we developed a novel reporter system to accurately measure NMD activity in individual cells. A genome-wide CRISPR-Cas9 knockout screen using this reporter system identified novel NMD-promoting factors, including multiple components of the SF3B complex and other U2 spliceosome factors. Interestingly, cells with mutations in the spliceosome genes SF3B1 and U2AF1, which are commonly found in myelodysplastic syndrome (MDS) and cancers, have overall attenuated NMD activity. Compared to wild type cells, SF3B1 and U2AF1 mutant cells were more sensitive to NMD inhibition, a phenotype that is accompanied by elevated DNA replication obstruction, DNA damage, and chromosomal instability. Remarkably, the sensitivity of spliceosome mutant cells to NMD inhibition was rescued by overexpression of RNase H1, which removes R-loops in the genome. Together, these findings shed new light on the functional interplay between NMD and RNA splicing and suggest a novel synthetic lethal strategy for the treatment of MDS and cancers with spliceosome mutations.

The aim of this study is to determine the demographic, pathological, and treatment-related factors that predict recurrence and survival in a Trinidadian cohort of breast cancer patients.The inclusion criteria for this study were female, over 18 years, and with a primary breast cancer diagnosis confirmed by a biopsy report occurring between 2010 and 2015 at Sangre Grande Hospital, Trinidad. Univariate associations with 5-year recurrence-free survival and 5-year overall survival were calculated using the Kaplan-Meier method for categorical variables and Cox Proportional Hazards for continuous variables. A multivariate model for prediction of recurrence and survival was determined using Cox regression.For the period 2010-2015, 202 records were abstracted. Five-year overall survival and recurrence-free survival rates were found to be 74.3% and 56.4%, respectively. Median times from first suspicious finding to date of biopsy report, date of surgery, and date of chemotherapy were 63 days, 125 days, and 189 days, respectively. In the univariate analysis, age (p = 0.038), stage (p 0.001), recurrence (p = 0.035), surgery (p = 0.016), ER (p 0.001) status, PR status (p 0.001), and subtype (p 0.001) were significantly associated with survival. Additionally, stage (p = 0.004), N score (p = 0.002), ER (p = 0.028) status, PR (p = 0.018) status, and subtype (p = 0.025) were significantly associated with recurrence. In the Cox multivariate model, Stage 4 was a significant predictor of survival (HR 6.77, 95% CI [0.09-2.49], p = 0.047) and N3 score was a significant predictor of recurrence (HR 4.47, 95% CI [1.29-15.54], p = 0.018).This study reports a 5-year breast cancer survival rate of 74.3%, and a recurrence-free survival rate of 56.4% in Trinidad for the period 2010-2015.

Background Cancer is the second leading cause of death in the Caribbean, including the islands of Trinidad and Tobago (TT). The population of TT consists of over 1.3 million people with diverse ancestral and sociocultural backgrounds, both of which may influence cancer incidence and mortality. The objective of this study was to examine incidence and mortality patterns and trends in TT. Methods Cancer surveillance data on 29,512 incident cancer cases reported to the Dr. Elizabeth Quamina Cancer Registry (population-based cancer registry of TT) between 1995 and 2009 were analyzed. Age-standardized rates, overall and by sex, ancestry, and geography, were reported. Results The highest incidence and mortality rates were observed for cancers related to reproductive organs in women, namely, breast, cervical, and uterine cancers, and prostate, lung and colorectal cancers among men. Average incidence rates were highest in areas covered by the Tobago Regional Health Authority (TRHA) (188 per 100,000), while average mortality rates were highest in areas covered by the North West Regional Health Authority (108 per 100,000). Nationals of African ancestry exhibited the highest rates of cancer incidence (243 per 100,000) and mortality (156 per 100,000) compared to their counterparts who were of East Indian (incidence, 125 per 100,000; mortality, 66 per 100,000) or mixed ancestry (incidence, 119 per 100,000; mortality, 66 per 100,000). Conclusions Our findings highlight the need for national investment to improve the understanding of the epidemiology of cancer in Trinidad and Tobago, and to ultimately guide much needed cancer prevention and control initiatives in the near future. Electronic supplementary material The online version of this article (10.1186/s12885-018-4625-x) contains supplementary material, which is available to authorized users.

Purpose In Trinidad and Tobago (TT), prostate cancer (CaP) is the most commonly diagnosed malignancy and the leading cause of cancer deaths among men. TT currently has one of the highest CaP mortality rates in the world. Methods 6,064 incident and 3,704 mortality cases of CaP occurring in TT from January 1995 to 31 December 2009 reported to the Dr. Elizabeth Quamina Cancer population-based cancer registry for TT, were analyzed to examine CaP survival, incidence, and mortality rates and trends by ancestry and geography. Results The age-standardized CaP incidence and mortality rates (per 100,000) based on the 1960 world-standardized in 2009 were 64.2 and 47.1 per 100,000. The mortality rate in TT increased between 1995 (37.9 per 100,000) and 2009 (79.4 per 100,000), while the rate in the US decreased from 37.3 per 100,000 to 22.1 per 100,000 over the same period. Fewer African ancestry patients received treatment relative to those of Indian and mixed ancestry (45.7%, 60.3%, and 60.9%, respectively). Conclusions Notwithstanding the limitations surrounding data quality, our findings highlight the increasing burden of CaP in TT and the need for improved surveillance and standard of care. Our findings highlight the need for optimized models to project cancer rates in developing countries like TT. This study also provides the rationale for targeted screening and optimized treatment for CaP to ameliorate the rates we report. Electronic supplementary material The online version of this article (10.1007/s10552-018-1038-8) contains supplementary material, which is available to authorized users.

Small nucleolar RNAs (snoRNAs) are noncoding RNAs that contribute to ribosome biogenesis and RNA splicing by modifying ribosomal RNA and spliceosome RNAs, respectively. We optimized a next-generation sequencing approach and a custom analysis pipeline to identify and quantify expression of snoRNAs in acute myeloid leukemia (AML) and normal hematopoietic cell populations. We show that snoRNAs are expressed in a lineage- and development-specific fashion during hematopoiesis. The most striking examples involve snoRNAs located in 2 imprinted loci, which are highly expressed in hematopoietic progenitors and downregulated during myeloid differentiation. Although most snoRNAs are expressed at similar levels in AML cells compared with CD34+, a subset of snoRNAs showed consistent differential expression, with the great majority of these being decreased in the AML samples. Analysis of host gene expression, splicing patterns, and whole-genome sequence data for mutational events did not identify transcriptional patterns or genetic alterations that account for these expression differences. These data provide a comprehensive analysis of the snoRNA transcriptome in normal and leukemic cells and should be helpful in the design of studies to define the contribution of snoRNAs to normal and malignant hematopoiesis.

To examine the factors associated with gynecologic cancer mortality risks, to estimate the mortality-to-incidence rate ratios (MIR) in Trinidad and Tobago (TT), and to compare the MIRs to those of select countries. Data on 3,915 incident gynecologic cancers reported to the National Cancer Registry of TT from 1 January 1995 to 31 December 2009 were analyzed using proportional hazards models to determine factors associated with mortality. MIRs for cervical, endometrial, and ovarian cancers were calculated using cancer registry data (TT), GLOBOCAN 2012 incidence data, and WHO Mortality Database 2012 data (WHO regions and select countries). Among the 3,915 incident gynecologic cancers diagnosed in TT during the study period, 1,795 (45.8%) were cervical, 1,259 (32.2%) were endometrial, and 861 (22.0%) were ovarian cancers. Older age, African ancestry, geographic residence, tumor stage, and treatment non-receipt were associated with increased gynecologic cancer mortality in TT. Compared to GLOBOCAN 2012 data, TT MIR estimates for cervical (0.49 vs. 0.53), endometrial (0.61 vs. 0.65), and ovarian cancers (0.32 vs. 0.48) were elevated. While the Caribbean region had intermediate gynecologic cancer MIRs, MIRs in TT were among the highest of the countries examined in the Caribbean region. Given its status as a high-income economy, the relatively high gynecologic cancer MIRs observed in TT are striking. These findings highlight the urgent need for improved cancer surveillance, screening, and treatment for these (and other) cancers in this Caribbean nation.

Highlights • Patient history and clinical findings are often not specific enough to arrive at a diagnosis of clear cell gallbladder carcinoma. • Consideration of morphologic and immunophenotypic features are essential to establish a diagnosis of clear cell gallbladder carcinoma. • It is important for clinicians to differentiate gallbladder cancer from metastases that most commonly arise from the kidneys and other possible secondary foci., Introduction Clear cell gallbladder carcinoma accounts for less than 1% of all gallbladder malignancies and demonstrates its unique histopathological characteristics in patients with no prior medical illness or familial predisposition. Presentation of case Here we present a case of a 56-year-old female, with no prior medical conditions presented with a 2-month history of upper abdominal pain. Routine hematological and biochemical tests were unremarkable. An abdominal ultrasound revealed the presence of a gallbladder calculi, and a fundic mass while magnetic resonance cholangiopancreatography revealed a 8.0 cm × 3.5 cm gallbladder mass. Computed tomography imaging excluded any distant haematogenous metastases. An open cholecystectomy with lymphadenectomy was proceeded by staging laparoscopy. Upon pathologic investigation, the morphologic and immunophenotypic features supported a diagnosis of clear cell variant of gallbladder carcinoma. Discussion Pathological prognostications for primary clear cell gall bladder carcinomas are not well defined due to the rarity of cases and possible misidentification as secondary metastases. Foci of adenocarcinoma within the tumor along with immunohistochemical staining probes can be informative in consideration of differential diagnosis. Conclusion In these cases, clinical case management should be personalized for increased survival with the possible incorporation of next generation sequencing approaches to guide therapeutic algorithms. We discuss this exceedingly rare case of the clear cell variant of gallbladder carcinoma in detail, highlighting some of the diagnostic, and clinical challenges.

Highlights • Phyllodes tumors are rare biphasic fibroepithelial neoplasms of breast and are classified as benign, borderline and malignant. • Resection of the tumor is the preferred treatment, whereas the role of radiation therapy, and adjuvant chemotherapy are undefined. • Wide excision or mastectomy should be performed with the goal of histological clear margins. • Axillary nodal dissection is not required., Introduction Phyllodes tumors (PTs) of the breast are extremely rare accounting for less than 1% of all breast tumors globally. Case records at the Trinidad and Tobago Cancer Registry show that only 0.003% of the reported breast cancer cases between 1995 and 2009 were PTs. Presentation of the case We report a 45-year-old woman who presented with swelling of the left breast. Ultrasound, mammogram and computed tomography imaging confirmed the presence of a mass in the right upper inner quadrant of the left breast. A biopsy revealed features supportive of a benign phyllodes tumor. A wide local excision was performed with the removal of a 19 × 11 × 10 cm mass. Histopathological analysis revealed features consistent with malignant phyllodes tumor. A complete mastectomy of the left breast was subsequently performed. Follow up over a 5-year period did not reveal any evidence of local recurrence or residual disease. To the best of our knowledge, this is the first case report of a malignant PT from the Caribbean and Latin America. Discussion Phyllodes tumors are classified as benign, borderline, or malignant based on histologic features including presence of a clear margin, cellularity, stromal overgrowth, tumor necrosis and mitotic index. The clinical challenge is to assess the risk of local tumor and metastatic recurrence in the context of fluid classifications. Conclusion Our case management approach shows that for patients with malignant PT, a thorough preoperative workup regimen followed by appropriate surgical intervention can result in a desirable prognosis.

Highlights • Malignant melanoma of the heel is a rare melanoma subtype globally. • This is the first reported case study of malignant melanoma of the heel in the Caribbean. • Our case highlights the need for the development of strategies to increase patient adherence in developing countries. • The literature search shows that this is the first reported case of malignant melanoma of the heel among non-fair skin individuals in 40 years. • Increasing rates of malignant melanoma of the heel suggest the need for increased public awareness, early diagnosis and shared clinical decision making., Introduction Malignant melanoma of the heel is a rare melanoma subtype with incidence rates that reflect the complex relationship between sun exposure at certain geographic locations, individual melanin levels and overall melanoma risk. It is oftentimes characterized by poor prognosis because of delays in presentation resulting in longitudinal tumor invasion, lymph node involvement and metastasis. Presentation of case A 59-year-old woman was admitted to the Eric Williams Medical Sciences Complex, Trinidad and Tobago with a 5 mm pruritic lesion on her left heel. At presentation, the lesion was asymmetric with border irregularities, color heterogeneity, with dynamics in elevation and overall size. She was subsequently diagnosed with malignant melanoma with left inguinal lymphadenopathy. A single stage wide local excision (WLE) of the left heel lesion with a split-thickness skin graft (STSG) and a left inguinal lymphadenectomy were performed. Dacarbazine (Bayer) was administered post operatively. Discussion Globally, the incidence of malignant melanoma is rapidly increasing, particularly, in countries like Trinidad and Tobago with a significant population of non-fair skinned individuals. There is need for strategic initiatives to increase patient adherence in these populations. Conclusion The rarity of malignant heel melanomas heightens the need for increased patient awareness and greater clinical surveillance to ensure early diagnosis and treatment.

Splenic artery aneurysms (SAAs) are an extremely rare cause of asymptomatic massive lower gastrointestinal bleeding with less than a handful of patients surviving such a presentation. A 24-year-old female presented in shock after multiple episodes of massive rectal bleeding. Imaging revealed a heterogeneous mass arising from the tail of the pancreas eroding into the splenic flexure of the colon. Further episodes of bleeding led to an exploratory laparotomy. Intraoperatively, a suspected neoplastic process arising from the tail of the pancreas with contiguous involvement of the splenic flexure of the colon and the greater curvature of the stomach was noted. Distal pancreaticosplenectomy, gastric wedge resection with segmental colectomy and primary anastomosis were performed. Histology revealed a SAA with rupture into the colon. This case report shows that en-bloc resection of a ruptured SAA can be performed with success in the emergency setting.

Giant pheochromocytomas are rare silent entities that do not present with the classical symptoms commonly seen in catecholamine-secreting tumors. In many cases they are accidentally discovered. The algorithm to diagnose a pheochromocytoma consists of biochemical evaluation and imaging of a retroperitoneal mass. The female patient in this case report presented with a palpable abdominal mass and was cured with surgical resection. She suffered no recurrence or complications on follow-up. The left retroperitoneal mass measured 27 × 18 × 12 cm and weighed 3,315 grams. Biochemical, radiological, and pathological examinations confirmed the diagnosis of a pheochromocytoma. In this paper, we report on our experience treating this patient and provide a summary of all giant pheochromocytomas greater than 10 cm reported to date in English language medical journals. Our patient’s giant cystic pheochromocytoma was the fourth heaviest and fifth largest maximal diameter identified using our literature search criteria. Additionally, this tumor had the largest maximal diameter of all histologically confirmed benign/low metastatic risk pheochromocytomas. Giant cystic pheochromocytomas are rare entities requiring clinical suspicion coupled with strategic diagnostic evaluation to confirm the diagnosis.

A situational analysis of breast cancer (BC) early detection services was carried out to investigate whether Trinidad and Tobago (T&T) has the framework for successful organized national screening. An online survey was designed to assess the availability, accessibility, quality control and assurance (QC&A), and monitoring and evaluation (M&E) mechanisms for public and private BC early detection. A focus group with local radiologists (n = 3) was held to identify unaddressed challenges and make recommendations for improvement. Major public hospitals offer free detection services with wait times of 1–6 months for an appointment. Private institutions offer mammograms for TTD$240 (USD$37) at minimum with same day service. Both sectors report a lack of trained staff. Using 1.2 mammograms per 10,000 women ≥40 years as sufficient, the public sector’s rate of 0.19 mammograms per 10,000 women ≥40 years for screening and diagnosis is inadequate. Program M&E mechanisms, QC&A guidelines for machinery use, delays in receipt of pathology reports, and unreliable drug access are further unaddressed challenges. T&T must first strengthen its human and physical resources, implement M&E and QC&A measures, strengthen cancer care, and address other impediments to BC early detection before investing in nationally organized BC screening.

Highlights • This case series provides details on one surgeon’s experience performing pelvic exenterations (PEs) in a developing country. • We provide a detailed surgical approach as well as complications based on the Clavien-Dindo classification. • We offer suggestions for surgeons in developing countries planning to undertake PEs., Introduction Pelvic exenteration (PE) is an ultra-radical surgical procedure characterized by the en bloc resection of the pelvic organs. Methods In this case series, we report retrospectively on four patients who underwent PE in Trinidad and Tobago from 2012 to 2016. One male patient had rectal cancer while one each of three women had cervical, colon, or rectal cancer. Results Early postoperative complications (≤30 days) occurred in all patients, while late complications (>30 days) occurred in one patient (Grade 1 – Clavien-Dindo classification). Disease recurrence occurred in 50% of patients, and the median overall survival was 8 months (range, 4–15 months). Discussion There are many inherent challenges to conducting such major procedures in developing countries, including inadequate blood product supplies, intensive care unit beds, and pre- and post-operative support services. With increased surgical capacity and support infrastructure, hospitals in these regions would be equipped to perform PEs with better outcomes. Conclusion This case series adds to existing data on the feasibility of performing PE in developing countries. We demonstrate that PE can be performed without major postoperative complications in a resource-limited hospital. To the best of our knowledge, this is the first case series that describes PE in the Caribbean.

The integrity of the p53 tumor suppressor pathway is compromised in the majority of cancers. In 7% of cancers p53 is inactivated by abnormally high levels of MDM2--an E3 ubiquitin ligase that polyubiquitinates p53, marking it for degradation. MDM2 engages p53 through its hydrophobic cleft, and blockage of that cleft by small molecules can re-establish p53 activity. Small molecule MDM2 inhibitors have been developed, but there is likely to be a high cost and long time period before effective drugs reach the market. An alternative is to repurpose FDA-approved drugs. This report describes a new approach, called Computational Conformer Selection, to screen for compounds that potentially inhibit MDM2. This screen was used to computationally generate up to 600 conformers of 3244 FDA-approved drugs. Drug conformer similarities to 41 computationally-generated conformers of MDM2 inhibitor nutlin 3a were ranked by shape and charge distribution. Quantification of similarities by Tanimoto combo scoring resulted in scores that ranged from 0.142 to 0.802. In silico docking of drugs to MDM2 was used to calculate binding energies and to visualize contacts between the top-ranking drugs and the MDM2 hydrophobic cleft. We present 15 FDA-approved drugs predicted to inhibit p53/MDM2 interaction.

Background: Progressive accumulation of misfolded or unfolded proteins is a symbol of impaired proteostasis and proteotoxicity. Such a chronic proteotoxicity is amenable to cell types that are post mitotically matured with lack of further differentiation or proliferation. Our recent discovery using a mouse model of familial human cardiac disease displayed protuberant shift in the redox state towards reductive stress (RS) in association with accumulation of toxic protein aggregates. Further, sustained trans-activation of Nrf2/antioxidant signaling caused RS in the myopathy hearts. Accordingly, we hypothesized that whether profound activation of Nrf2/antioxidant signaling and subsequent RS may cause pathological remodeling in cardiomyocyte. The aim of this study was to investigate the effect of sustained pharmacological activation of Nrf2 on cardiac remodeling. Methods: HL1 cardiomyocytes were used as an in vitro model to study the RS-mediated cardiac remodeling. They were treated with 2-10 μM of potential Nrf2-inducers; sulforaphane (SF), di-methyl fumarate (DMF) and novel small molecules (C-38, C-50, C-63 and C-66) to establish RS by sustained activation of Nrf2/antioxidant signaling. Next, we investigated the implications of RS in cardiomyocyte remodeling by analyzing transcriptional and translational mechanisms using immunoblotting, qPCR, immunofluorescence, GSH and NADPH redox measurements in HL1 cells. Results: Dose dependent effects for individual small molecules including known Nrf2 inducers (SF and DMF) revealed distinct pro-reductive and reductive intracellular (i.e. reductive stress) environments. In fact, the obligatory activation of Nrf2 signaling was associated with significant upregulation of antioxidant enzymes and small molecular thiols including glutathione (GSH). Surprisingly, while pro-reductive condition in HL1 cells was subdued, the RS induced cardiomyocyte hypertrophy was evident from microscopic examination and molecular signature (increased expression of ANF and BNF) after 24-48 hrs of Nrf2 activation. Conclusion: In summary, the chemical induced sustained activation of Nrf2 leading to formation of reductive stress showed hypertrophic remodeling in HL1 cardiomyocytes.

Breast cancer (BC) is the most common newly diagnosed cancer among women in Trinidad and Tobago (TT) and BC mortality rates are among the highest in the world. Globally, racial/ethnic trends in BC incidence, mortality and survival have been reported. However, such investigations have not been conducted in TT, which has been noted for its rich diversity. In this study, we investigated associations among ancestry, geography and BC incidence, mortality and survival in TT. Data on 3767 incident BC cases, reported to the National Cancer Registry of TT, from 1995 to 2007, were analyzed in this study. Women of African ancestry had significantly higher BC incidence and mortality rates (Incidence: 66.96; Mortality: 30.82 per 100,000) compared to women of East Indian (Incidence: 41.04, Mortality: 14.19 per 100,000) or mixed ancestry (Incidence: 36.72, Mortality: 13.80 per 100,000). Geographically, women residing in the North West Regional Health Authority (RHA) catchment area followed by the North Central RHA exhibited the highest incidence and mortality rates. Notable ancestral differences in survival were also observed. Women of East Indian and mixed ancestry experienced significantly longer survival than those of African ancestry. Differences in survival by geography were not observed. In TT, ancestry and geographical residence seem to be strong predictors of BC incidence and mortality rates. Additionally, disparities in survival by ancestry were found. These data should be considered in the design and implementation of strategies to reduce BC incidence and mortality rates in TT.

Advances in human genetics and genomic sciences and the corresponding explosion of biomedical technologies have deepened current understanding of human health and revolutionized medicine. In developed nations, this has led to marked improvements in disease risk stratification and diagnosis. These advances have also led to targeted intervention strategies aimed at promoting disease prevention, prolonging disease onset, and mitigating symptoms, as in the well-known case of breast cancer and the BRCA1 gene. In contrast, in the developing nation of Trinidad and Tobago, this scientific revolution has not translated into the development and application of effective genomics-based interventions for improving public health. While the reasons for this are multifactorial, the underlying basis may be rooted in the lack of pertinence of internationally driven genomics research to the local public health needs in the country, as well as a lack of relevance of internationally conducted genetics research to the genetic and environmental contexts of the population. Indeed, if Trinidad and Tobago is able to harness substantial public health benefit from genetics/genomics research, then there is a dire need, in the near future, to build local capacity for the conduct and translation of such research. Specifically, it is essential to establish a national human genetics/genomics research agenda in order to build sustainable human capacity through education and knowledge transfer and to generate public policies that will provide the basis for the creation of a mutually beneficial framework (including partnerships with more developed nations) that is informed by public health needs and contextual realities of the nation.

We describe the presentation, treatment, clinical outcome, and targeted genome analysis of a metastatic salivary acinic cell carcinoma (AciCC). A 71-year-old male presented with a 3 cm right tail of a parotid lesion, first detected as a nodule by the patient seven months earlier. He had a right total parotidectomy with cranial nerve VII resection, right facial nerve resection and grafting, resection of the right conchal cartilage, and right modified radical neck dissection. The primary tumor revealed AciCC with two distinct areas: a well-differentiated component with glandular architecture and a dedifferentiated component with infiltrative growth pattern associated with prominent stromal response, necrosis, perineural invasion, and cellular pleomorphism. Tumor staging was pT4 N0 MX. Immunohistochemistry staining showed pankeratin (+), CD56 (−), and a Ki67 proliferation index of 15%. Upon microscopic inspection, 49 local lymph nodes resected during parotidectomy were negative for cancer cells. Targeted sequencing of the primary tumor revealed deletions of CDKN2A and CDKN2B, a nonsense mutation in ARID2, and single missense mutations of unknown significance in nine other genes. Despite postoperative localized radiation treatment, follow-up whole body PET/CT scan showed lung, soft tissue, bone, and liver metastases. The patient expired 9 months after resection of the primary tumor.

Trinidad and Tobago (T&T) has one of the highest breast cancer (BC) mortality rates in the Americas. For the period 1995-2007, 5-year BC survival rates in T&T were approximately 30%. One possible explanation for T&T’s low BC survival rates could be late presentation and detection. Currently, opportunistic BC screening programs exist in the public and private sectors and some advocate for an organized national BC screening program. However, the World Health Organization (WHO) only recommends the implementation of organized BC screening in high resource settings. Thus, the purpose of this study was to conduct a situational analysis of breast cancer early detection services in the public and private sector to identify gaps in service and determine if a national organized screening program is warranted. The quality, availability, affordability and accessibility of BC screening services in the public and private sector was assessed with an online survey using the Cancer Control WHO Guide for Effective Programs framework. The inter-rater reliability and validity was assessed during a pilot phase at two public institutions. All public and private facilities and NGOs with BC early detection services were invited to participate in the survey. A focus group with key radiologists in T&T was held to review the results of the study, identify unaddressed challenges and make recommendations for improvement. Significant challenges exist in service availability and access, physical and human resources, program monitoring and evaluation, quality control mechanisms, follow-up diagnostics and care. Specifically, services are largely monopolized by the private sector at a cost to nationals. Public services which are free, are restricted to major hospitals where wait times extend to 3-6 months, thus contributing to a disparity in access. Both sectors cite a lack of trained staff such as specialist breast radiographers and radiologists. There is also a lack of preventative maintenance and investment in new machinery. No quality control guidelines or manuals for machinery use exist in the public sector. Across both sectors, 70% of institutions have no program evaluation. Other systemic problems such as long wait times for pathology reports also undermine the efficacy of current BC early detection efforts. An organized national breast cancer screening program is not recommended for T&T at this time. More pressing concerns across the cancer care continuum need to be first addressed particularly in strengthening human and physical resource capacity and service quality for detection, diagnosis, and follow-up care. Also, research into factors such as time-to-treat, frequency of late stage presentation and misdiagnosis warrant further attention if T&T is to improve its BC survival rate. Note: This abstract was not presented at the meeting. Citation Format: Kimberly Badal, Fidel Rampersad, Hamish Mohammed, Murrie Moosoodeen, Siva Konduru, Nalini Kokaram Maharaj, Adaila Russel, Melissa Nathan, Marisa Nimrod, Wayne A. Warner, Rajini Haraksingh, Adetunji Toriola. Challenges to breast cancer early detection in the developing, high income country of Trinidad and Tobago [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4226. doi:10.1158/1538-7445.AM2017-4226

An increasing number of industrial, household, drug, and cosmetic chemicals are developed and marketed each year. Most new compounds to which workers and consumers may be exposed, especially topically, are screened for safety by standardized skin-irritation toxicity testing protocols. However, the testing of these materials for their phototoxic potential is not routine, and thus the development of phototoxicity test procedures has been slow. Today there exists a need for meaningful, reliable, and standardized tests for determining the phototoxic potential of the many chemicals and products that we may come in contact with in our daily lives.Phototoxicity (photoirritation) is a light-induced skin response, similar to an exaggerated sunburn, that can be elicited after a single exposure to a photoactive chemical. This chemical may reach the target tissues directly by topical application, or indirectly by ingestion, or by other means that allow the chemical to enter the bloodstream. Phototoxicity differs from...

Small nucleolar RNAs (snoRNAs) are small (90-300nt) non-coding guide RNAs found in all multicellular organisms. H/ACA and CD box snoRNAs localize to the nucleolar region and are part of a catalytic multicomponent protein complex selecting target RNAs based on complementarity. They are responsible for the site-specific pseudouridylation and 2' O methylation of ribosomal RNAs, respectively. ScaRNAs localize to the Cajal body and are responsible for the methylation and pseudouridylation of splicesomal RNAs U1, U2, U4, U5, and U12. There are also orphan snoRNAs without any known RNA targets. Recent studies have suggested an expanded role for snoRNAs outside of ribosomal biogenesis, including the regulation of RNA splicing and chromatin remodeling. Moreover, emerging data suggest that aberrantly expressed snoRNAs may contribute to neoplastic transformation. Here, we characterize snoRNA expression in normal human hematopoiesis and in AML. We first developed a novel strategy to identify and quantify properly processed snoRNAs. This is important, since standard RNA sequencing or array-based analyses cannot distinguish between processed snoRNAs and primary mRNA transcripts of the host genes (most snoRNAs are contained in the introns of coding genes). In brief, we take advantage of the fact that, like miRNAs, snoRNAs contain a free 3'-hydroxyl group, which allows for efficient ligation to sequencing adaptors. Following size selection (20-200 nt) to enrich for small non-coding RNAs, the libraries are sequenced on the Illumina next generation sequencing platform. We also have developed a novel analysis pipeline that maps areas of contiguous alignment in the genome, forming ab initio "clusters" representing snoRNAs. Using this sequencing assay, we first interrogated snoRNA expression in hematopoietic cells from healthy individuals. Specifically, we sequenced small RNA libraries derived from CD34+ cells, promyelocytes, neutrophils, monocytes, T-cells, and B-cells. Of the 269 known snoRNAs, 132 (49%) were expressed in one or more hematopoietic cell population. Likewise, 80 of 112 (71%) of known H/ACA box snoRNAs, and all 21 scaRNAs were expressed. In addition, we identified (and computationally validated using SnoReport, snoGPS, and an in-house snoFinder script) 8 putative novel snoRNAs (1 H/ACA box and 7 CD box snoRNAs). Most snoRNAs were stably expressed across all hematopoietic lineages. However, there were numerous examples of snoRNAs that were specifically enriched in a specific hematopoietic cell population (e.g., CD34+ cells). We also identified several snoRNAs that were up- or down-regulated during granulocytic differentiation. For example, many of the orphan C/D box snoRNAs contained in the imprinted DLK/DIO3 locus on chromosome 14q32 are significantly down-regulated during granulocytic differentiation. To determine whether snoRNAs are frequently dysregulated in AML, we next sequenced small non-coding RNAs isolated from the bone marrow of 33 patients with de novo acute myeloid leukemia (all with a normal karyotype). Using a strict 5% false discovery rate, only 9.3% of CD box snoRNAs and 0.9% of H/ACA box snoRNAs were found to have significantly increased or decreased expression compared with normal CD34+ cells (including some of the putative novel snoRNAs). Of note, no differentially expressed snoRNAs were detected comparing AML with or without DNMT3A mutations or with or without IDH1/2 mutations. We next interrogated published whole genome sequencing data to determine whether there were any cytogenetically silent genetic alterations in snoRNAs. No recurring point mutations or small insertions/deletions were detected in snoRNA genes in 50 cases of AML. In summary, we developed a new next-generation sequencing approach and analysis pipeline to quantify snoRNAs. We show that, compared with coding genes, snoRNA expression is more stable across different hematopoietic lineages, consistent with a housekeeping function. However, examples of developmentally-regulated and lineage-restricted snoRNA expression were identified. Finally, we show that a small subset of snoRNAs appear to be dysregulated in AML, although genetic alterations the specifically target snoRNAs appear to be rare in AML. Disclosures No relevant conflicts of interest to declare.

Background: Breast cancer (BC) is clearly a major global public health concern. This concern is more evident in developing countries like Trinidad and Tobago (TT) where BC mortality rates are among the highest in the Caribbean and the world. This twin island nation has a population of 1,328,019 of multicultural ancestries including East Indian (35%), African (34%) and mixed ancestry (23%). TT is the most industrialized of all Caribbean nations and offers comprehensive, no-cost healthcare to all citizens. Five Regional Health Authorities (RHAs) are responsible for the provision of healthcare services, including oncology care. Given the high BC mortality rates documented in TT, we sought to examine whether differences in BC incidence, mortality and/or survival existed by race/ethnicity or geography. Methods: This study analyzed the associations among race/ethnicity and geographical residence (based on RHA) and BC incidence and mortality as well as survival of 3777 female BC cases reported between January 1995 and December 2005 to the Dr. Elizabeth Quamina Cancer Registry, which serves as the National Cancer Registry of TT. The population data was based on the census figures for 2000 and 2010 as reported by the Central Statistical Office of Trinidad and Tobago. Chi-square tests were used to examine the associations between race/ethnicity and geographical residence and BC incidence and mortality. Lifetest analyses were used to examine differences in BC survival by race/ethnicity and geographical area of residence. Results: Overall 22.7% of the cases were diagnosed at age 60 years. The largest proportion of BC diagnosed at 60 years was observed among women of African ancestry (p Conclusion: These findings demonstrate that in TT, BC incidence and mortality rates differed significantly by race/ethnicity and geographical residence, while BC survival differed by race/ethnicity. These findings highlight the importance of targeted cancer prevention and control efforts to address the observed disparities in BC in TT. Citation Format: Wayne A. Warner, Yee Lam Lee, Shelina Ramnarine, Simeon Slovacek, Veronica Roach, Matthew Ellis, Adana Llanos. Associations among race/ethnicity, geography, and breast cancer incidence and mortality in Trinidad and Tobago. [abstract]. In: Proceedings of the Seventh AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 9-12, 2014; San Antonio, TX. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2015;24(10 Suppl):Abstract nr B19.

Advances in human genetics and genomic sciences and the corresponding explosion of biomedical technologies have deepened current understanding of human health and revolutionized medicine. In developed nations, this has led to marked improvements in disease risk stratification and diagnosis. These advances have also led to targeted intervention strategies aimed at promoting disease prevention, prolonging disease onset, and mitigating symptoms, as in the well-known case of breast cancer and the BRCA1 gene. In contrast, in the developing nation of Trinidad and Tobago, this scientific revolution has not translated into the development and application of effective genomics-based interventions for improving public health. While the reasons for this are multifactorial, the underlying basis may be rooted in the lack of pertinence of internationally driven genomics research to the local public health needs in the country, as well as a lack of relevance of internationally conducted genetics research to the genetic and environmental contexts of the population. Indeed, if Trinidad and Tobago is able to harness substantial public health benefit from genetics/genomics research, then there is a dire need, in the near future, to build local capacity for the conduct and translation of such research. Specifically, it is essential to establish a national human genetics/genomics research agenda in order to build sustainable human capacity through education and knowledge transfer and to generate public policies that will provide the basis for the creation of a mutually beneficial framework (including partnerships with more developed nations) that is informed by public health needs and contextual realities of the nation.

Highlights • This is the first reported right accessory spleen laying retro-duodenal reported. • This is the second largest accessory spleen reported. • Accessory spleen is a possibility in a retroperitoneal tumor differential diagnosis. • We present a literature review of the other right accessory spleen cases., Introduction Unlike left sided accessory spleen that are seen in 10–30% of cases at autopsy, cases of right accessory spleens are extremely rare. This congenital body of healthy splenic tissue simulates tumors from neighboring organs and presents a challenge in formulating a differential diagnosis. Presentation of case We present the case of a patient whose CT scan of the abdomen showed a large mass, 11 × 8 cm, arising retro-duodenal and lying just anterior to the right kidney. To the best of our knowledge, this is the only case where the accessory spleen was found retro-duodenal, directly anterior to the kidney and completely separate from the supra-renal gland. The chief complaint of the patient was right upper quadrant pain, radiating to the back, and colicky in nature. The patient was diagnosed with duodenal gastro-intestinal stromal tumor and a retro-peritoneal sarcoma. The mass was removed via a Kocher’s incision and immunohistological examination showed that it was a right sided accessory spleen. The patient’s left sided spleen appeared normal. Discussion Efforts to distinguish an accessory spleen from a retroperitoneal tumor with available scans, percutaneous biopsy or biochemical tests are inconclusive. Differential diagnosis between a retroperitoneal tumor and an accessory spleen can only be made after surgical exploration. Conclusion This case highlights the fact that surgeons should consider the possibility of an accessory spleen when making a differential diagnosis of retroperitoneal tumors.

Massively parallel DNA and RNA sequencing approaches have generated data on thousands of breast cancer genomes. In this review, we consider progress largely from the perspective of new concepts and hypotheses raised so far. These include challenges to the multistep model of breast carcinogenesis and the discovery of new defects in DNA repair through sequence analysis. Issues for functional genomics include the development of strategies to differentiate between mutations that are likely to drive carcinogenesis and bystander background mutations, as well as the importance of mechanistic studies that examine the role of mutations in genes with roles in splicing, histone methylation, and long non-coding RNA function. The application of genome-annotated patient-derived breast cancer xenografts as a potentially more reliable preclinical model is also discussed. Finally, we address the challenge of extracting medical value from genomic data. A weakness of many datasets is inadequate clinical annotation, which hampers the establishment of links between the mutation spectra and the efficacy of drugs or disease phenotypes. Tools such as dGene and the DGIdb are being developed to identify possible druggable mutations, but these programs are a work in progress since extensive molecular pharmacology is required to develop successful ‘genome-forward’ clinical trials. Examples are emerging, however, including targeting HER2 in HER2 mutant breast cancer and mutant ESR1 in ESR1 endocrine refractory luminal-type breast cancer. Finally, the integration of DNA- and RNA-based sequencing studies with mass spectrometry-based peptide sequencing and an unbiased determination of post-translational modifications promises a more complete view of the biochemistry of breast cancer cells and points toward a new discovery horizon in our understanding of the pathophysiology of this complex disease. Electronic supplementary material The online version of this article (doi:10.1186/s13058-014-0460-4) contains supplementary material, which is available to authorized users.