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Cervical cancer is one of the most common cancers in women. Despite progress in prevention through Human Papilloma Virus (HPV) vaccination and success in early detection of cervical cancer through cytologic screening and HPV detection, there remains an unequal cervical cancer burden in low-resource settings, both in developed and developing countries. We have previously shown that the CervicalMethDx test can provide a Cervical Intraepithelial Neoplasia (CIN) grade 2-3 risk score, by assessing DNA methylation in a panel of three human genes (ZNF516, FKBP6 and INTS1) in samples from the United States (US), Puerto Rico, and Chile. We now tested the performance of the CervicalMethDx test on HPV-positive CIN2 and CIN3 cases (n=113) from Honduras collected from participants in the ESTAMPA clinical trial (NCT01881659). ESTAMPA (EStudio multicéntrico de TAMizaje y triaje de cáncer de cuello uterino con pruebas del virus del PApiloma humano; Spanish acronym) is a multicentric study of cervical cancer screening with HPV testing and assessment of triage methods in Latin America, which has accrued close to 50,000 participants from twelve recruitment centers in nine Latin American countries since 2013. We hypothesized that the CervicalMethDx test can identify HPV positive women most likely to be diagnosed with CIN grades 2 and 3 by anatomic pathologists, before they are referred to colposcopy-driven biopsies. We assessed DNA methylation by quantitative Real Time Methylation Specific PCR (qMSP) analysis of sodium bisulfite-modified genomic DNA. Primers and probes were previously designed to specifically amplify the promoters of the 3 genes of interest and the promoter of a reference gene, β-actin, to assess DNA input. We performed blinded retrospective studies on well-characterized clinical samples in PreservCyt sample transport media (ThinPrep, Hologic), comparing DNA methylation levels in samples from Honduras and 88 HPV-positive previously tested US samples with No Intraepithelial Lesions or Malignancy (NILM), as controls. Our results showed that the CervicalMethDx test can correctly classify 96% of CIN2 (n=62) samples with 92% Sensitivity, 98% Specificity, and an AUC of 0.96 as well as 97% of CIN3 (n=51) samples with 96% Sensitivity, 98% Specificity, and an AUC of 0.97. Moreover, the assay correctly classified 96% of CIN2-CIN3 samples combined (n=113) with 95% Sensitivity, 98% Specificity, and AUC of 0.97, when compared to samples with NILM (n=88). Our results suggest that the CervicalMethDx test is a new and valuable tool to stratify HPV positive women prior to colposcopy-driven biopsies in developed countries and ablative treatment in developing countries, most of which are unnecessary. These results warrant further evaluation of the CervicalMethDX test in prospective, population-based studies, assessing the use of precision DNA methylation algorithms to triage HPV positive women before referral to colposcopy-driven biopsies or ablative treatments in cervical cancer screening clinics world-wide. Citation Format: Laura Palmieri, Fernando Zamuner, Yessy Cabrera, Jafet Ortiz-Quintero, Dieila Giomo De Lima, Ana Purcell-Wiltz, Amanda García-Negrón, Ashley Ramos-López, Mariana Brait, David Sidransky, Annabelle Ferrera, Rafael E. Guerrero-Preston. A precision DNA methylation test to triage HPV positive women before referral to colposcopy-driven biopsies or ablative treatment in cervical cancer screening clinics worldwide [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB107.

Cervical cancer is one of the most common cancers in women. Despite progress in prevention through Human Papilloma Virus [HPV] vaccination and success in early detection of cervical cancer through cytologic screening and HPV detection, there remains an unequal burden in low-resource settings in developed and developing countries. Applying novel methylation-based detection methods, we validated a panel of three human methylated genes (ZNF516, FKBP6 and INTS1) on discarded cervical liquid cytology samples from clinical laboratories in the United States during the development of the CervicalMethDx test. We hypothesized that the CervicalMethDx test can identify HPV positive women most likely to be diagnosed with Cervical Intraepithelial Neoplasia (CIN) grades 2 and 3 by anatomic pathologists, before they are referred to colposcopy-driven biopsies. We assessed DNA methylation by Quantitative Real Time Methylation Specific PCR (QMSP) analysis of sodium bisulfite-modified genomic DNA. Primers and probes were previously designed to specifically amplify the promoters of the 3 genes of interest and the promoter of a reference gene, β-actin, to assess DNA input. We performed blinded retrospective studies on well-characterized, discarded, HPV-positive clinical samples in PreservCyt sample transport media (Thin Prep, Hologic), comparing DNA methylation levels in samples from CIN2 and CIN3 cases (346), and 120 controls without evidence of Intraepithelial Lesions or Malignancy (NILM) as an endpoint. Our results showed that the CervicalMethDx test can correctly classify 96% of CIN2 (n=197) samples with 93% Sensitivity, 100% Specificity, and an AUC of 0.96 as well as 95% of CIN3 (n=149) samples with 91% Sensitivity, 100% Specificity, and an AUC of 0.96. Moreover, the assay correctly classified 94% of CIN2-CIN3 samples combined (n=346) with 92% Sensitivity, 100% Specificity, an AUC of 0.96, and a 100% positive predictive value (PPV), when compared to samples with NILM (n=120). The CervicalMethDx test high PPV supports the addition of this low-cost molecular test to cervical cancer screening algorithms worldwide. Our results suggest that the CervicalMethDx test is a new and valuable tool to stratify HPV positive women prior to colposcopy-driven biopsies in developed countries and ablative treatment in developing countries, most of which are unnecessary. These results warrant further evaluation of the CervicalMethDX test in prospective, population-based studies, assessing the use of precision DNA methylation algorithms to triage women before colposcopy-driven biopsies or ablative treatments worldwide. Citation Format: Laura Palmieri, Fernando T. Zamuner, Dieila Giomo De Lima, Keerthana Gosala, Eli Winkler, Yash Prashar, Ana Purcell-Wiltz, Amanda García-Negrón, Ashley Ramos-Lopez, David Sidransky, Mariana Brait, Rafael Guerrero-Preston. CervicalMethDx: A precision DNA methylation test to identify advanced disease risk in cervical cancer screening algorithms. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4192.

Cervical cancer incidence is 32% higher among Latina women in the United States (US) and 78% higher among women in Puerto Rico compared to non-Latina White (NLW) women, yet it is largely preventable through screening. Latina women with public insurance or uninsured are significantly less likely to have had a Pap test, undergo fewer Pap and high-Risk Human Papilloma Virus (HPV) testing than NLW and Black women, and are less likely than women of other races/ethnicities to return for recommended follow-up after an abnormal Pap test. Utilizing the PRECEDE-PROCEED model, Community Based Participatory Research strategies and the NIH I-Corps hypothesis-driven research method of customer discovery, we implemented the Community Genomic Health Promotion Program in Federally Qualified Health Centers to evaluate cervical cancer screening needs in six rural municipalities in Puerto Rico. We recently validated in clinical liquid cytology samples that the CervicalMethDx test, a precision DNA methylation test, can identify HPV positive women most likely to be diagnosed with Cervical Intraepithelial Neoplasia (CIN) grades 2 and 3. We hypothesized that a molecular pathology test which identifies advanced premalignant cervical lesions in urine, can be distributed by commercial channels targeting populations with low screening rates. The CervicalMethDx quantitative Methylation Specific PCR (qMSP) test was used to quantify ZNF516, FKBP6 and INTS1 promoter DNA methylation in well-characterized, discarded, liquid cytology clinical samples from Puerto Rico. We then tested the CervicalMethDx test concordance on paired cytobrush (n=82) and urine samples (n=82) from Chile. The CervicalMethDx test correctly classified 86% of discarded liquid cytology clinical samples from HPV-positive women, when comparing DNA methylation in CIN2/CIN3 samples (n=47) to samples with a cervical pathology diagnosis of No Intraepithelial Lesions or Malignancy (n=18), with 83% Sensitivity, 94% Specificity, Area Under the Curve (AUC) of 0.88, and 98% positive predictive value (PPV). The concordance of the CervicalMethDx test in paired brush and urine samples was moderate (rho=0.45). The Wilcoxon signed-rank test showed no statistically significant differences for the CervicalMethDx test in paired brush and urine samples (p=0.14) These encouraging preliminary results show the successful identification of advanced premalignant cervical lesions in liquid cytology media and urine samples from Latina women. The data suggests that urine should be explored as a biofluid for cervical cancer screening. Future research may lead to expanded screening options, enabling women with low access to health care (public insurance or uninsured) to collect a specimen at home, and either send it to a testing facility or perform the test at home. Citation Format: Carmen Ili, Priscilla Brebi, Jaime López, Fernando T. Zamunér, Ana Purcell-Wiltz, Amanda García-Negrón, Keilyn Vale-Lassalle, Keerthana Gosala, Mirla Otero-García, Camila Rivero-Maldonado, Teresa Torres-Rivera, Adriana Pérez-Vicente, Ashley Ramos-López, David Sidransky, Mariana Brait, Rafael E. Guerrero-Preston. Community Genomic Health Promotion Program evaluates precision DNA methylation biomarkers for use in self-testing and collection kits in urine, to increase cervical cancer screening rates among Latina women [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1978.

Carbonized lignin has been proposed as a sustainable and domestic source of activated, amorphous, graphitic, and nanostructured carbon for many industrial applications as the structure can be tuned through processing conditions. However, the inherent variability of lignin and its complex physicochemical structure resulting from feedstock and pulping selection make the Process-Structure-Property-Performance (PSPP) relationships hard to define. In this work, radial distribution functions (RDFs) from synchrotron X-ray and neutron scattering of lignin-based carbon composites (LBCCs) are investigated using the Hierarchical Decomposition of the Radial Distribution Function (HDRDF) modelling method to characterize the local atomic environment and develop quantitative PSPP relationships. PSPP relationships for LBCCs defined by this work include crystallite size dependence on lignin feedstock as well as increasing crystalline volume fraction, nanoscale composite density, and crystallite size with increasing reduction temperature.

OBJECTIVE: Guillain-Barré syndrome (GBS) is an uncommon autoimmune disorder that follows infection or vaccination, and increased incidence has been reported during Zika virus (ZIKV) transmission. During the 2016 ZIKV epidemic, the Puerto Rico Department of Health (PRDH) implemented the Enhanced GBS Surveillance System (EGBSSS). Here, we describe EGBSSS implementation and evaluate completeness, validity, and timeliness. METHODS: GBS cases were identified using passive surveillance and discharge diagnostic code for GBS. Completeness was evaluated by capture-recapture methods. Sensitivity and positive predictive value (PPV) for confirmed GBS cases were calculated for both case identification methods. Median time to completion of key time steps were compared by quarter (Q1–4) and hospital size. RESULTS: A total of 122 confirmed GBS cases with onset of neurologic illness in 2016 were identified. Capture-recapture methodology estimated that four confirmed GBS cases were missed by both identification methods. Identification of cases by diagnostic code had a higher sensitivity than passive surveillance (89% vs. 80%), but a lower PPV (60% vs. 72%). There was a significant decrease from Q1 to Q3 in median time from hospital admission to case reporting (11 days vs. 2 days, p = 0.032) and from Q2 to Q3 in median time from specimen receipt to arbovirus laboratory test reporting (35 days vs. 26 days, p = 0.004). CONCLUSION: EGBSSS provided complete, valid, and increasingly timely surveillance data, which guided public health action and supported healthcare providers during the ZIKV epidemic. This evaluation provides programmatic lessons for GBS surveillance and emergency response surveillance. [P R Health Sci J 2018;37(Special Issue):S85-S92]