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OBJECTIVE: In Haiti, cervical cancer continues to cause high levels of mortality and morbidity due to lack of resources and political unrest. Haitian women employed in factories are especially vulnerable because they are unable to take time away from work to access health resources. We aimed to describe a low-cost intervention which successfully addressed this need. METHODS: We present a retrospective review of data gathered through a public-private partnership, in which women working in garment factories near Port-au-Prince, Haiti, were offered health education, clinical breast exam, and free human papillomavirus (HPV) self-swab testing at their place of employment. Women testing positive for HPV were subsequently tested using visual inspection with acetic acid (VIA) to inform treatment referrals, and treated with mobile thermocoagulation in factory infirmaries. Factory-employed healthcare workers were trained on cancer screening, including VIA and clinical breast exam. RESULTS: A total of 6843 out of 6983 (98%) female factory employees attended free reproductive health education sessions, and 4005 out of 4153 eligible women (97%) were screened using HPV self-swab testing; 5176 women received a clinical breast exam. Of the women screened for HPV, 1001 (25%) tested positive and 905 (90%) of HPV-positive women received VIA testing and thermocoagulation. The intervention had a total cost of US$76 000, over half of which was spent on an HPV testing machine. CONCLUSIONS: Innovative approaches to the prevention of cervical cancer are especially necessary in very low-resource, politically unstable environments like Haiti. Self-swab and screen-and-treat programs in the workplace were acceptable to employees and factory owners. This low-cost model was reached vulnerable women through a public-private partnership, and tracked them through screening and treatment. It could be implemented elsewhere or extended to include other health services.
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As the SARS-CoV-2 virus spread throughout the world, millions of positive cases of COVID-19 were registered and, even though there are millions of people already vaccinated against SARS-CoV-2, a large part of the global population remains vulnerable to contracting the virus. Massive nasopharyngeal sample collection in Puerto Rico at the beginning of the pandemic was limited by the scarcity of trained personnel and testing sites. To increase SARS-CoV-2 molecular testing availability, we evaluated the diagnostic accuracy of self-collected nasal, saliva, and urine samples using the TaqPath reverse transcription polymerase chain reaction (RT-PCR) COVID-19 kit to detect SARS-CoV-2. We also created a colorimetric loop-mediated isothermal amplification (LAMP) laboratory developed test (LDT) to detect SARS-CoV-2, as another strategy to increase the availability of molecular testing in community-based laboratories. Automated RNA extraction was performed in the KingFisher Flex instrument, followed by PCR quantification of SARS-CoV-2 on the 7500 Fast Dx RT-PCR using the TaqPath RT-PCR COVID-19 molecular test. Data was interpreted by the COVID-19 Interpretive Software from Applied Biosystems and statistically analyzed with Cohen's kappa coefficient (k). Cohen's kappa coefficient (k) for paired nasal and saliva samples showed moderate agreement (0.52). Saliva samples exhibited a higher viral load. We also observed 90% concordance between LifeGene-Biomarks' SARS-CoV-2 Rapid Colorimetric LAMP LDT and the TaqPath RT-PCR COVID-19 test. Our results suggest that self-collected saliva is superior to nasal and urine samples for COVID-19 testing. The results also suggest that the colorimetric LAMP LDT is a rapid alternative to RT-PCR tests for the detection of SARS-CoV-2. This test can be easily implemented in clinics, hospitals, the workplace, and at home; optimizing the surveillance and collection process, which helps mitigate global public health and socioeconomic upheaval caused by airborne pandemics.
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COVID-19 , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , SARS-CoV-2 , Saliva , Manejo de Especímenes , Humanos , Saliva/virología , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , COVID-19/diagnóstico , COVID-19/virología , COVID-19/orina , Técnicas de Amplificación de Ácido Nucleico/métodos , Manejo de Especímenes/métodos , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , ARN Viral/análisis , ARN Viral/orina , ARN Viral/genética , ARN Viral/aislamiento & purificación , Prueba de Ácido Nucleico para COVID-19/métodos , Sensibilidad y Especificidad , Puerto Rico/epidemiología , Prueba de COVID-19/métodosRESUMEN
Differentially methylated regions (DMRs) can be used as head and neck squamous cell carcinoma (HNSCC) diagnostic, prognostic and therapeutic targets in precision medicine workflows. DNA from 21 HNSCC and 10 healthy oral tissue samples was hybridized to a genome-wide tiling array to identify DMRs in a discovery cohort. Downstream analyses identified differences in promoter DNA methylation patterns in oral, laryngeal and oropharyngeal anatomical regions associated with tumor differentiation, nodal involvement and survival. Genome-wide DMR analysis showed 2,565 DMRs common to the three subsites. A total of 738 DMRs were unique to laryngeal cancer (n=7), 889 DMRs were unique to oral cavity cancer (n=10) and 363 DMRs were unique to pharyngeal cancer (n=6). Based on the genome-wide analysis and a Gene Ontology analysis, 10 candidate genes were selected to test for prognostic value and association with clinicopathological features. TIMP3 was associated with tumor differentiation in oral cavity cancer (P=0.039), DAPK1 was associated with nodal involvement in pharyngeal cancer (P=0.017) and PAX1 was associated with tumor differentiation in laryngeal cancer (P=0.040). A total of five candidate genes were selected, DAPK1, CDH1, PAX1, CALCA and TIMP3, for a prevalence study in a larger validation cohort: Oral cavity cancer samples (n=42), pharyngeal cancer tissues (n=25) and laryngeal cancer samples (n=52). PAX1 hypermethylation differed across HNSCC anatomic subsites (P=0.029), and was predominantly detected in laryngeal cancer. Kaplan-Meier survival analysis (P=0.043) and Cox regression analysis of overall survival (P=0.001) showed that DAPK1 methylation is associated with better prognosis in HNSCC. The findings of the present study showed that the HNSCC subsites oral cavity, pharynx and larynx display substantial differences in aberrant DNA methylation patterns, which may serve as prognostic biomarkers and therapeutic targets.
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INTRODUCTION: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic revealed a worldwide lack of effective molecular surveillance networks at local, state, and national levels, which are essential to identify, monitor, and limit viral community spread. SARS-CoV-2 variants of concern (VOCs) such as Alpha and Omicron, which show increased transmissibility and immune evasion, rapidly became dominant VOCs worldwide. Our objective was to develop an evidenced-based genomic surveillance algorithm, combining reverse transcription polymerase chain reaction (RT-PCR) and sequencing technologies to quickly identify highly contagious VOCs, before cases accumulate exponentially. METHODS: Deidentified data were obtained from 508,969 patients tested for coronavirus disease 2019 (COVID-19) with the TaqPath COVID-19 RT-PCR Combo Kit (ThermoFisher) in four CLIA-certified clinical laboratories in Puerto Rico (n = 86,639) and in three CLIA-certified clinical laboratories in the United States (n = 422,330). RESULTS: TaqPath data revealed a frequency of S Gene Target Failure (SGTF) > 47% for the last week of March 2021 in both, Puerto Rico and US laboratories. The monthly frequency of SGTF in Puerto Rico steadily increased exponentially from 4% in November 2020 to 47% in March 2021. The weekly SGTF rate in US samples was high (>8%) from late December to early January and then also increased exponentially through April (48%). The exponential increase in SGFT prevalence in Puerto Rico was concurrent with a sharp increase in VOCs among all SARS-CoV-2 sequences from Puerto Rico uploaded to Global Influenza Surveillance and Response System (GISAID) (n = 461). Alpha variant frequency increased from <1% in the last week of January 2021 to 51.5% of viral sequences from Puerto Rico collected in the last week of March 2021. CONCLUSIONS: According to the proposed evidence-based algorithm, approximately 50% of all SGTF patients should be managed with VOCs self-quarantine and contact tracing protocols, while WGS confirms their lineage in genomic surveillance laboratories. Our results suggest this workflow is useful for tracking VOCs with SGTF.
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COVID-19 , SARS-CoV-2 , Secuencia de Bases , COVID-19/diagnóstico , COVID-19/epidemiología , Humanos , Medicina de Precisión , SARS-CoV-2/genética , Estados Unidos/epidemiologíaRESUMEN
AIM: The aim of the study was to evaluate the use of global and gene-specific DNA methylation changes as potential biomarkers for gallbladder cancer (GBC) in a cohort from Chile. MATERIAL & METHODS: DNA methylation was analyzed through an ELISA-based technique and quantitative methylation-specific PCR. RESULTS: Global DNA Methylation Index (p = 0.02) and promoter methylation of SSBP2 (p = 0.01) and ESR1 (p = 0.05) were significantly different in GBC when compared with cholecystitis. Receiver curve operator analysis revealed promoter methylation of APC, CDKN2A, ESR1, PGP9.5 and SSBP2, together with the Global DNA Methylation Index, had 71% sensitivity, 95% specificity, a 0.97 area under the curve and a positive predictive value of 90%. CONCLUSION: Global and gene-specific DNA methylation may be useful biomarkers for GBC clinical assessment.
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Colecistitis/diagnóstico , Metilación de ADN , Neoplasias de la Vesícula Biliar/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Línea Celular Tumoral , Chile , Colecistitis/genética , Femenino , Neoplasias de la Vesícula Biliar/genética , Humanos , Masculino , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular , Regiones Promotoras Genéticas , Curva ROC , Análisis de Secuencia de ADNRESUMEN
Tumor suppressor genes (TSGs) are commonly inactivated by somatic mutation and/or promoter methylation; yet, recent high-throughput genomic studies have not identified key TSGs inactivated by both mechanisms. We pursued an integrated molecular analysis based on methylation binding domain sequencing (MBD-seq), 450K Methylation arrays, whole exome sequencing, and whole genome gene expression arrays in primary head and neck squamous cell carcinoma (HNSCC) tumors and matched uvulopalatopharyngoplasty tissue samples (UPPPs). We uncovered 186 downregulated genes harboring cancer specific promoter methylation including PAX1 and PAX5 and we identified 10 key tumor suppressor genes (GABRB3, HOXC12, PARP15, SLCO4C1, CDKN2A, PAX1, PIK3AP1, HOXC6, PLCB1, and ZIC4) inactivated by both promoter methylation and/or somatic mutation. Among the novel tumor suppressor genes discovered with dual mechanisms of inactivation, we found a high frequency of genomic and epigenomic alterations in the PAX gene family of transcription factors, which selectively impact canonical NOTCH and TP53 pathways to determine cell fate, cell survival, and genome maintenance. Our results highlight the importance of assessing TSGs at the genomic and epigenomic level to identify key pathways in HNSCC, deregulated by simultaneous promoter methylation and somatic mutations.
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Carcinoma de Células Escamosas/genética , Metilación de ADN , Silenciador del Gen , Genes Supresores de Tumor , Neoplasias de Cabeza y Cuello/genética , Regiones Promotoras Genéticas , Carcinoma de Células Escamosas/metabolismo , Línea Celular Tumoral , Estudios de Cohortes , Islas de CpG , Femenino , Neoplasias de Cabeza y Cuello/metabolismo , Humanos , Masculino , Mutación , Factor de Transcripción PAX5/genética , Factor de Transcripción PAX5/metabolismo , Factores de Transcripción Paired Box/genética , Factores de Transcripción Paired Box/metabolismo , Receptor Notch1/genética , Receptor Notch1/metabolismo , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Cervical cancer is a major health concern among women in Latin America due to its high incidence and mortality. Therefore, the discovery of molecular markers for cervical cancer screening and triage is imperative. The aim of this study was to use a genome wide DNA methylation approach to identify novel methylation biomarkers in cervical cancer. DNA from normal cervical mucosa and cervical cancer tissue samples from Chile was enriched with Methylated DNA Immunoprecipitation (MeDIP), hybridized to oligonucleotide methylation microarrays and analyzed with a stringent bioinformatics pipeline to identify differentially methylated regions (DMRs) as candidate biomarkers. Quantitative Methylation Specific PCR (qMSP) was used to study promoter methylation of candidate DMRs in clinical samples from two independent cohorts. HPV detection and genotyping were performed by Reverse Line Blot analysis. Bioinformatics analysis revealed GGTLA4, FKBP6, ZNF516, SAP130, and INTS1 to be differentially methylated in cancer and normal tissues in the Discovery cohort. In the Validation cohort FKBP6 promoter methylation had 73% sensitivity and 80% specificity (AUC = 0.80). ZNF516 promoter methylation was the best biomarker, with both sensitivity and specificity of 90% (AUC = 0.92), results subsequently corroborated in a Prevalence cohort. Together, ZNF516 and FKBP6 exhibited a sensitivity of 84% and specificity of 81%, when considering both cohorts. Our genome wide DNA methylation assessment approach (MeDIP-chip) successfully identified novel biomarkers that differentiate between cervical cancer and normal samples, after adjusting for age and HPV status. These biomarkers need to be further explored in case-control and prospective cohorts to validate them as cervical cancer biomarkers.
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Metilación de ADN , Papillomavirus Humano 18 , Regiones Promotoras Genéticas , Proteínas de Unión a Tacrolimus/genética , Neoplasias del Cuello Uterino/genética , Dedos de Zinc , Adulto , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Chile , Femenino , Genoma Humano , Humanos , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteínas de Unión a Tacrolimus/metabolismo , Neoplasias del Cuello Uterino/etnología , Neoplasias del Cuello Uterino/virologíaRESUMEN
Cervical cancer is a leading cause of cancer deaths in women worldwide and infection by high-risk human papillomavirus types is a precursor event. The cellular FLICE-like inhibitory protein (c-FLIP) has been found to be overexpressed in several types of cancers and could be associated with cervical cancer progression because of its ability to inhibit the apoptotic process. To detect c-FLIP expression in cervical cancer, an immunohistochemical staining was performed, using tissue microarrays, on a series of 536 archival biopsy samples, including normal cervical tissues, low-grade and high-grade squamous intraepithelial lesions, and squamous cervical carcinomas. The epithelium in the normal cervix and low-grade squamous intraepithelial lesions mainly stained negatively for c-FLIP, whereas high-grade intraepithelial lesions and cancer samples showed an elevated expression of c-FLIP. A direct association was observed between the increasing grade of the lesion and the intensity of c-FLIP staining, in which the frequency of intense c-FLIP expression increased from 12.5% in the normal tissue to 82.1% in the cervical cancer tissue. An increased expression of c-FLIP may be an important factor in the progression of cervical cancer. This finding could aid in identifying patients with preneoplastic lesions at greater risk of developing cervical cancer. c-FLIP expression in cervical tissue may be a potential cervical cancer progression marker.
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Biomarcadores de Tumor/análisis , Proteína Reguladora de Apoptosis Similar a CASP8 y FADD/biosíntesis , Carcinoma de Células Escamosas/metabolismo , Displasia del Cuello del Útero/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proteína Reguladora de Apoptosis Similar a CASP8 y FADD/análisis , Carcinoma de Células Escamosas/mortalidad , Carcinoma de Células Escamosas/patología , Progresión de la Enfermedad , Femenino , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Persona de Mediana Edad , Análisis de Matrices Tisulares , Neoplasias del Cuello Uterino/mortalidad , Neoplasias del Cuello Uterino/patología , Adulto Joven , Displasia del Cuello del Útero/mortalidad , Displasia del Cuello del Útero/patologíaRESUMEN
Racial, ethnic and class disparities in cancer incidence and mortality have been well documented. Disparities in the utilization of preventive, curative and treatment services among ethnic minorities have been reported. Screening can be effective at detecting cancer at treatable stages, but a large proportion of people at risk have not been screened or are not regularly screened, as recommended by the American Cancer Society's national guidelines. Early detection technologies have the potential of both influencing mortality from cancer, as well as enhancing primary prevention through detection and removal of lesions that could potentially develop into cancer. Cancer is an epigenetic disease characterized by the breakdown of DNA methylation and histones modification patterns. Epigenetic approaches may contribute to a reduction in cancer health disparities impacting early detection and increasing cancer treatment options. Epigenetic events represent important mechanism(s) by which gene function is selectively activated or inactivated, through genetic and non-genetic manifestations. Emerging evidence indicates that various epigenetic alterations, such as global histones modifications and DNA hypomethylation, common to most types of cancer, are modified by environmental exposures throughout the life course. A simple, easily explained and easy to understand non-invasive test, such as the DNA methylation index, that may screen for several cancer sites at once, may remove some of the existing barriers to cancer screening utilization, and contribute to the reduction of cancer disparities. Epigenetic approaches may also prove to be useful in identifying environmental and lifestyle factors that contribute to the prevalence of other chronic conditions in high risk populations, such as Puerto Rican populations in the United States and Puerto Rico.
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Pruebas Genéticas/métodos , Disparidades en el Estado de Salud , Neoplasias/genética , Epigénesis Genética , Humanos , Neoplasias/epidemiología , Factores de RiesgoRESUMEN
INTRODUCTION: Two hundred and thirty nine (239) drinking water systems in Puerto Rico are not connected to the Puerto Rico Aqueducts and Sewers Authority (PRASA), and are thus known as Non-PRASA drinking water systems. Population served estimates by Non-PRASA systems are in the 100,000 to 300,000 range. OBJECTIVES: To identify the determinants of compliance with drinking water standards by rural drinking water systems in Puerto Rico. To identify the best analytical methods for studying the problem of non-compliance with drinking water standards in Puerto Rico and its generalization to similar communities elsewhere. METHODS: We reviewed capacity development and drinking water system evaluations performed by governmental and academic institutions between 1993 and 2004. Community and system variables were used to fit a multilevel model to predict compliance with drinking water standards. Data was obtained from the Environmental Protection Agency' Safe Drinking Water Information System and the Puerto Rico Health Department drinking water database for 231 systems, serving 90,000 persons. RESULTS: There was an 11% increase in compliance (1996 = 4%; 2000=15%), a decrease of 13,634 people served by non-compliant systems (1996 = 86,169; 2000 = 72,535) and a 6% decrease in the number of non-compliant systems which had installed treatment equipment (1996 = 93%; 2000=87%). The prevalence of compliance among those systems that had installed treatment equipment was higher than among those systems that did not have treatment equipment, after adjusting by the time period (est. POR = 2.2, 95% CI, 1.40 - 3.44). CONCLUSIONS: Our findings suggest alternative public health strategies are needed to ensure sustained safe water capacity in rural communities.
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Adhesión a Directriz/estadística & datos numéricos , Abastecimiento de Agua/normas , Puerto Rico , Salud RuralRESUMEN
INTRODUCTION: Pharmaceutical residues and other organic wastewater contaminants (OWC) have been shown to survive conventional water-treatment processes and persist in potable water supplies. OBJECTIVE: To estimate the geographical distribution of the Predicted Environmental Concentration (PEC) of selected drugs prescribed by office based physicians in the United States (US), after non-metabolized residues have been excreted and processed in wastewater treatment plants. METHODS: The geographical distribution of the PEC in surface waters of pharmaceutical residues was calculated, in four regions of the US. Prescription drug data was obtained from the National Ambulatory Medical Care Survey (NAMCS). The PEC of three drugs prescribed by office based physicians in the US between 1998 and 2000 was compared to the concentrations of these pharmaceuticals found in a surface water characterization project conducted by the United States Geological Survey between 1999 and 2000. RESULTS: There were 803,185,420 medications prescribed by office-based physicians in the US between 1998 and 2000. Relief of pain, hormonal, cardiovascular and antimicrobial medications followed very similar prescription patterns, both in terms of quantity and geographical distribution. Together these four types of medications account for more than half of the medications prescribed between 1998 and 2000. The concentration of pharmaceutical residues found in the drinking water supply was not significantly correlated to the PEC of pharmaceuticals prescribed by office-based physicians. CONCLUSION: The geographical distribution of medications prescribed by office based physicians in the US underlines the need to implement effective public health strategies.