NCI SEER-Linked Virtual Tissue Repository Pilot.

BACKGROUND: The Surveillance, Epidemiology, and End Results (SEER) Program with the National Cancer Institute tested whether population-based cancer registries can serve as honest brokers to acquire tissue and data in the SEER-Linked Virtual Tissue Repository (VTR) Pilot. METHODS: We collected formalin-fixed, paraffin-embedded tissue and clinical data from patients with pancreatic ductal adenocarcinoma (PDAC) and breast cancer (BC) for two studies comparing cancer cases with highly unusual survival (≥5 years for PDAC and ≤30 months for BC) to pair-matched controls with usual survival (≤2 years for PDAC and ≥5 years for BC). Success was defined as the ability for registries to acquire tissue and data on cancer cases with highly unusual outcomes. RESULTS: Of 98 PDAC and 103 BC matched cases eligible for tissue collection, sources of attrition for tissue collection were tissue being unavailable, control paired with failed case, second control that was not requested, tumor necrosis ≥20%, and low tumor cellularity. In total, tissue meeting the study criteria was obtained for 70 (71%) PDAC and 74 (72%) BC matched cases. For patients with tissue received, clinical data completeness ranged from 59% for CA-19-9 after treatment to >95% for margin status, whether radiation therapy and chemotherapy were administered, and comorbidities. CONCLUSIONS: The VTR Pilot demonstrated the feasibility of using SEER cancer registries as honest brokers to provide tissue and clinical data for secondary use in research. Studies using this program should oversample by 45% to 50% to obtain sufficient sample size and targeted population representation and involve subspecialty matter expert pathologists for tissue selection.

Perceptions of Preventable Cancer Burden Among US-Based Firefighters: A Mixed Methods Cross-Sectional Study.

OBJECTIVE: Characterize experiences with cancer and perceptions of the preventable cancer burden attributable to diet among firefighters. DESIGN: A cross-sectional survey was distributed to assess cancer history and perceptions of cancer. SETTING: US. PARTICIPANTS: US-based firefighters. MAIN OUTCOME MEASURE(S): History of cancer, perceptions of cancer, and perceptions of diet as a means to prevent cancer. ANALYSIS: Descriptive statistics and use of the Behaviour Change Technique Taxonomy v1 (BCTTv1) to evaluate qualitative responses. RESULTS: A total of 471 firefighters participated. Nearly half (48.4%) voiced they strongly agreed that they were at risk for cancer, whereas 44.6% agreed that changing diet could decrease cancer risk. The most common BCTTv1 codes focused on types of education, including "Instruction on how to perform the behavior" (45.1%, n = 189), followed by those centered on behavior execution (eg, "Action planning" [24.8%, n = 104]). Qualitatively, many were concerned about misinformation. CONCLUSIONS AND IMPLICATIONS: Firefighters acknowledge the role of diet in cancer risk and have a desire for knowledge and behavioral support (eg, goal setting) that includes an emphasis on evidence and tackling misinformation. This information should serve as the basis of future interventions that target diet.

NKX2-2 based nuclei sorting on frozen human archival pancreas enables the enrichment of islet endocrine populations for single-nucleus RNA sequencing.

BACKGROUND: Current approaches to profile the single-cell transcriptomics of human pancreatic endocrine cells almost exclusively rely on freshly isolated islets. However, human islets are limited in availability. Furthermore, the extensive processing steps during islet isolation and subsequent single cell dissolution might alter gene expressions. In this work, we report the development of a single-nucleus RNA sequencing (snRNA-seq) approach with targeted islet cell enrichment for endocrine-population focused transcriptomic profiling using frozen archival pancreatic tissues without islet isolation. RESULTS: We cross-compared five nuclei isolation protocols and selected the citric acid method as the best strategy to isolate nuclei with high RNA integrity and low cytoplasmic contamination from frozen archival human pancreata. We innovated fluorescence-activated nuclei sorting based on the positive signal of NKX2-2 antibody to enrich nuclei of the endocrine population from the entire nuclei pool of the pancreas. Our sample preparation procedure generated high-quality single-nucleus gene-expression libraries while preserving the endocrine population diversity. In comparison with single-cell RNA sequencing (scRNA-seq) library generated with live cells from freshly isolated human islets, the snRNA-seq library displayed comparable endocrine cellular composition and cell type signature gene expression. However, between these two types of libraries, differential enrichments of transcripts belonging to different functional classes could be observed. CONCLUSIONS: Our work fills a technological gap and helps to unleash frozen archival pancreatic tissues for molecular profiling targeting the endocrine population. This study opens doors to retrospective mappings of endocrine cell dynamics in pancreatic tissues of complex histopathology. We expect that our protocol is applicable to enrich nuclei for transcriptomics studies from various populations in different types of frozen archival tissues.

Annual Report to the Nation on the Status of Cancer, part 2: Early assessment of the COVID-19 pandemic's impact on cancer diagnosis.

BACKGROUND: With access to cancer care services limited because of coronavirus disease 2019 control measures, cancer diagnosis and treatment have been delayed. The authors explored changes in the counts of US incident cases by cancer type, age, sex, race, and disease stage in 2020. METHODS: Data were extracted from selected US population-based cancer registries for diagnosis years 2015-2020 using first-submission data from the North American Association of Central Cancer Registries. After a quality assessment, the monthly numbers of newly diagnosed cancer cases were extracted for six cancer types: colorectal, female breast, lung, pancreas, prostate, and thyroid. The observed numbers of incident cancer cases in 2020 were compared with the estimated numbers by calculating observed-to-expected (O/E) ratios. The expected numbers of incident cases were extrapolated using Joinpoint trend models. RESULTS: The authors report an O/E ratio <1.0 for major screening-eligible cancer sites, indicating fewer newly diagnosed cases than expected in 2020. The O/E ratios were lowest in April 2020. For every cancer site except pancreas, Asians/Pacific Islanders had the lowest O/E ratio of any race group. O/E ratios were lower for cases diagnosed at localized stages than for cases diagnosed at advanced stages. CONCLUSIONS: The current analysis provides strong evidence for declines in cancer diagnoses, relative to the expected numbers, between March and May of 2020. The declines correlate with reductions in pathology reports and are greater for cases diagnosed at in situ and localized stage, triggering concerns about potential poor cancer outcomes in the coming years, especially in Asians/Pacific Islanders. PLAIN LANGUAGE SUMMARY: To help control the spread of coronavirus disease 2019 (COVID-19), health care organizations suspended nonessential medical procedures, including preventive cancer screening, during early 2020. Many individuals canceled or postponed cancer screening, potentially delaying cancer diagnosis. This study examines the impact of the COVID-19 pandemic on the number of newly diagnosed cancer cases in 2020 using first-submission, population-based cancer registry database. The monthly numbers of newly diagnosed cancer cases in 2020 were compared with the expected numbers based on past trends for six cancer sites. April 2020 had the sharpest decrease in cases compared with previous years, most likely because of the COVID-19 pandemic.

Impact on the Volume of Pathology Reports Before and During the COVID-19 Pandemic in SEER Cancer Registries.

INTRODUCTION: Health care procedures including cancer screening and diagnosis were interrupted due to the COVID-19 pandemic. The extent of this impact on cancer care in the United States is not fully understood. We investigated pathology report volume as a reflection of trends in oncology services pre-pandemic and during the pandemic. METHODS: Electronic pathology reports were obtained from 11 U.S. central cancer registries from NCI's SEER Program. The reports were sorted by cancer site and document type using a validated algorithm. Joinpoint regression was used to model temporal trends from January 2018 to February 2020, project expected counts from March 2020 to February 2021 and calculate observed-to-expected ratios. Results were stratified by sex, age, cancer site, and report type. RESULTS: During the first 3 months of the pandemic, pathology report volume decreased by 25.5% and 17.4% for biopsy and surgery reports, respectively. The 12-month O/E ratio (March 2020-February 2021) was lowest for women (O/E 0.90) and patients 65 years and older (O/E 0.91) and lower for cancers with screening (melanoma skin, O/E 0.86; breast, O/E 0.88; lung O/E 0.89, prostate, O/E 0.90; colorectal, O/E 0.91) when compared with all other cancers combined. CONCLUSIONS: These findings indicate a decrease in cancer diagnosis, likely due to the COVID-19 pandemic. This decrease in the number of pathology reports may result in a stage shift causing a subsequent longer-term impact on survival patterns. IMPACT: Investigation on the longer-term impact of the pandemic on pathology services is vital to understand if cancer care delivery levels continue to be affected.

Human nuclear hormone receptor activity contributes to malaria parasite liver stage development.

Chemical genetic approaches have had a transformative impact on discovery of drug targets for malaria but have primarily been used for parasite targets. To identify human pathways required for intrahepatic development of parasite, we implemented multiplex cytological profiling of malaria infected hepatocytes treated with liver stage active compounds. Some compounds, including MMV1088447 and MMV1346624, exhibited profiles similar to cells treated with nuclear hormone receptor (NHR) agonist/antagonists. siRNAs targeting human NHRs, or their signaling partners identified eight genes that were critical for Plasmodium berghei infection. Knockdown of NR1D2, a host NHR, significantly impaired parasite growth by downregulation of host lipid metabolism. Importantly, treatment with MMV1088447 and MMV1346624 but not other antimalarials, phenocopied the lipid metabolism defect of NR1D2 knockdown. Our data underlines the use of high-content imaging for host-cellular pathway deconvolution, highlights host lipid metabolism as a drug-able human pathway and provides new chemical biology tools for studying host-parasite interactions.

Exploring a Tetrahydroquinoline Antimalarial Hit from the Medicines for Malaria Pathogen Box and Identification of its Mode of Resistance as PfeEF2.

New antimalarial treatments with novel mechanism of action are needed to tackle Plasmodium falciparum infections that are resistant to first-line therapeutics. Here we report the exploration of MMV692140 (2) from the Pathogen Box, a collection of 400 compounds that was made available by Medicines for Malaria Venture (MMV) in 2015. Compound 2 was profiled in in vitro models of malaria and was found to be active against multiple life-cycle stages of Plasmodium parasites. The mode of resistance, and putatively its mode of action, was identified as Plasmodium falciparum translation elongation factor 2 (PfeEF2), which is responsible for the GTP-dependent translocation of the ribosome along mRNA. The compound maintains activity against a series of drug-resistant parasite strains. The structural motif of the tetrahydroquinoline (2) was explored in a chemistry program with its structure-activity relationships examined, resulting in the identification of an analog with 30-fold improvement of antimalarial asexual blood stage potency.

Disruption of miR-18a Alters Proliferation, Photoreceptor Replacement Kinetics, Inflammatory Signaling, and Microglia/Macrophage Numbers During Retinal Regeneration in Zebrafish.

In mammals, photoreceptor loss causes permanent blindness, but in zebrafish (Danio rerio), photoreceptor loss reprograms Müller glia to function as stem cells, producing progenitors that regenerate photoreceptors. MicroRNAs (miRNAs) regulate CNS neurogenesis, but the roles of miRNAs in injury-induced neuronal regeneration are largely unknown. In the embryonic zebrafish retina, miR-18a regulates photoreceptor differentiation. The purpose of the current study was to determine, in zebrafish, the function of miR-18a during injury-induced photoreceptor regeneration. RT-qPCR, in situ hybridization, and immunohistochemistry showed that miR-18a expression increases throughout the retina between 1 and 5 days post-injury (dpi). To test miR-18a function during photoreceptor regeneration, we used homozygous miR-18a mutants (miR-18ami5012), and knocked down miR-18a with morpholino oligonucleotides. During photoreceptor regeneration, miR-18ami5012 retinas have fewer mature photoreceptors than WT at 7 and 10 dpi, but there is no difference at 14 dpi, indicating that photoreceptor regeneration is delayed. Labeling dividing cells with 5-bromo-2'-deoxyuridine (BrdU) showed that at 7 and 10 dpi, there are excess dividing progenitors in both mutants and morphants, indicating that miR-18a negatively regulates injury-induced proliferation. Tracing 5-ethynyl-2'-deoxyuridine (EdU) and BrdU-labeled cells showed that in miR-18ami5012 retinas excess progenitors migrate to other retinal layers in addition to the photoreceptor layer. Inflammation is critical for photoreceptor regeneration, and RT-qPCR showed that in miR-18ami5012 retinas, inflammatory gene expression and microglia activation are prolonged. Suppressing inflammation with dexamethasone rescues the miR-18ami5012 phenotype. Together, these data show that in the injured zebrafish retina, disruption of miR-18a alters proliferation, inflammation, the microglia/macrophage response, and the timing of photoreceptor regeneration.

Micron-sized particle separation with standing surface acoustic wave-Experimental and numerical approaches.

Traditional cell/particle isolation methods are time-consuming and expensive and can lead to morphology disruptions due to high induced shear stress. To address these problems, novel lab-on-a-chip-based purification methods have been employed. Among various methods introduced for the separation and purification of cells and synthetics particles, acoustofluidics has been one of the most effective methods. Unlike traditional separation techniques carried out in clinical laboratories based on chemical properties, the acoustofluidic process relies on the physical properties of the sample. Using acoustofluidics, manipulating cells and particles can be achieved in a label-free, contact-free, and highly biocompatible manner. To optimize the functionality of the platform, the numerical study should be taken into account before conducting experimental tests to save time and reduce fabrication expenses. Most current numerical studies have only considered one-dimensional harmonic standing waves to simulate the acoustic pressure distribution. However, one-dimensional simulations cannot calculate the actual acoustic pressure distribution inside the microchannel due to its limitation in considering longitudinal waves. To address this limitation, a two-dimensional numerical simulation was conducted in this study. Our numerical simulation investigates the effects of the platform geometrical and operational conditions on the separation efficiency. Next, the optimal values are tested in an experimental setting to validate these optimal parameters and conditions. This work provides a guideline for future acoustofluidic chip designs with a high degree of reproducibility and efficiency.

Melanoma Tumor Depth Quality Audit: A Nonmatch Analysis.

Background: The National Cancer Institute's Surveillance Research Program (SRP) received reports from cancer registries in the Surveillance, Epidemiology, and End Results (SEER) Program concerning the coding of melanoma tumor depth. To address these concerns, SRP developed an algorithm to identify melanoma depth measurement values and conducted a nonmatch analysis. Methods: A nonmatch analysis was conducted on 1,117 cases diagnosed between 2010 and 2017. With the help of Information Management Services, a natural language processing algorithm was developed to identify melanoma tumor depth values along with a gold standard for comparison. A randomly sampled data set was created to compare the algorithm-generated and gold standard values to the originally reported values; these were analyzed using SAS software version 9.4. Analyses were conducted to determine the distribution of nonmatches by demographics and estimate the distribution of nonmatches by the derived T variable according to the 7th edition of the American Joint Committee on Cancer (AJCC)'s AJCC Cancer Staging Manual. Results: Of the 1,117 cases, 849 cases (76%) were a match between the originally reported values and the gold standard. The majority of cases were found to be in male patients (60%) and non-Hispanic White patients (93%). When comparing derived AJCC-7 T based on the originally reported value to the gold standard, 16% of the original derived AJCC-7 T values were incorrect, with most of the nonmatches resulting in incorrectly coding a case as TX instead of T1. Conclusion: In total, 24% of cases were found to have a discrepancy in the originally recorded values. Decimal errors made up 3% of all cases in this nonmatch analysis. This algorithm may prove to be an essential tool in optimizing registry resources by flagging inconsistencies via automated text review to be adjudicated by registrars, improving their quality of data as needed.

Quantification of Pain and Distress Associated With Intranasal Midazolam Administration in Children and Evaluation of Validity of Four Observational Measures.

OBJECTIVES: The aims of this study were to quantify the pain and distress associated with the administration of intranasal (IN) midazolam in young children using 4 observational measures and to evaluate the degree of validity of these measures. METHODS: We conducted a prospective observational pilot study. Children aged 1 to 7 years requiring IN midazolam were enrolled. Children were videotaped, and scores were assigned to baseline and administration phases using the Observational Scale of Behavioral Distress-Revised (OSBD-R), Children's Hospital of Eastern Ontario Pain Scale (CHEOPS), and the Faces-Legs-Activity-Cry-Consolability (FLACC) scale. The cry duration following administration was assessed. Interrater reliability and convergent validity were determined for all 4 measures. Internal consistency and responsivity for the OSBD-R, CHEOPS, and FLACC scales were determined. RESULTS: We enrolled 20 children. The mean OSBD-R, CHEOPS, and FLACC scores associated with administration of IN midazolam were 27.1 (SD, 13.5), 11.5 (SD, 1.2), and 8.9 (SD, 2.7), respectively. The mean cry duration was 105.5 (SD, 68.8) seconds. The intraclass correlation coefficients for all measures ranged from 0.82 to 0.99. The Cronbach α's for the OSBD-R, CHEOPS, and FLACC were between 0.71 and 0.97. Pearson correlation coefficients for comparisons between OSBD-R, CHEOPS, and FLACC were between 0.82 and 0.96 but were between 0.32 and 0.51 for comparisons involving cry duration. CONCLUSIONS: We have identified estimates of pain and distress associated with administration of IN midazolam in young children that can be used to determine desired effect sizes for trials that study interventions to treat this pain and distress. The OSBD-R, CHEOPS, and FLACC scales are suitable choices for outcome measures.

Body fatness and mTOR pathway activation of breast cancer in the Women's Circle of Health Study.

Energy imbalance has an important role in breast cancer prognosis. Hyperactive mechanistic Target of Rapamycin (mTOR) pathway is associated with breast tumor growth, but the extent to which body fatness is associated with mTOR pathway activities in breast cancer is unclear. We performed immunostaining for mTOR, phosphorylated (p)-mTOR, p-AKT, and p-p70S6K in tumor tissue from 590 women (464 African Americans/Blacks and 126 Whites) with newly diagnosed invasive breast cancer in the Women's Circle of Health Study. Anthropometric measures were taken by study staff, and body composition was measured by bioelectrical impedance analysis. Linear regressions were used to estimate percent differences in protein expression between categories of body mass index (BMI), waist circumference, waist/hip ratio, fat mass, fat mass index, and percent body fat. We observed that BMI ≥ 35.0 vs. <25 kg/m2 was associated with 108.3% (95% CI = 16.9%-270.9%) and 101.8% (95% CI = 17.0%-248.8%) higher expression in p-mTOR and normalized p-mTOR, i.e., p-mTOR/mTOR, respectively. Quartiles 4 vs. 1 of waist/hip ratio was associated with 41.8% (95% CI = 5.81%-89.9%) higher mTOR expression. Similar associations were observed for the body fat measurements, particularly in patients with estrogen receptor-negative (ER-) tumors, but not in those with ER+ tumors, although the differences in associations were not significant. This tumor-based study found positive associations between body fatness and mTOR pathway activation, evident by a p-mTOR expression, in breast cancer. Our findings suggest that mTOR inhibition can be a treatment strategy to prevent the recurrence of these tumors in obese individuals.

Parametric study on the geometrical parameters of a lab-on-a-chip platform with tilted planar electrodes for continuous dielectrophoretic manipulation of microparticles.

Advances in lab-on-a-chip (LOC) devices have led to significant improvements in the on-chip manipulation, separation, sorting, and isolation of particles and cells. Among various LOC-based approaches such as inertia-based methods, acoustophoresis, and magnetophoresis, the planar-slanted-electrode dielectrophoresis (DEP) method has demonstrated great potential as a label-free, cost-effective, and user-friendly approach. However, the devices built based on this method suffer from low flow throughput compared to devices functioning based on other LOC-based manipulation approaches. In order to overcome this obstacle, the geometrical parameters of these types of DEP-based devices must be studied to increase the effectiveness of DEP manipulation. With the consideration of both numerical and experimental studies, this paper studies the geometrical factors of a LOC platform consisting of tilted planar electrodes with the goal of achieving higher throughput in continuous manipulation of polystyrene particles. COMSOL Multiphysics software was used to study the effect of the electrodes geometry on the induced electric field. The simulation results show that by increasing the electrode's width and decreasing the electrode's spacing, higher DEP force is generated. Furthermore, the experimental outcomes indicated that lower channel height, higher voltage, and larger particle size resulted in the most improvement to DEP manipulation. Additionally, the experimental results demonstrated that slanted electrodes with an angle of 8° with respect to the direction of flow provide a more effective configuration.

WITHDRAWN: Comparison of Preadministered and Coadministered Lidocaine for Treating Pain and Distress Associated With Intranasal Midazolam Administration in Children: A Randomized Clinical Trial.

This article has been withdrawn at the request of the authors and editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at https://www.elsevier.com/about/policies/article-withdrawal.

Comparison of preadministered and coadministered lidocaine for treating pain and distress associated with intranasal midazolam administration in children: A randomized clinical trial.

OBJECTIVE: Pain and distress associated with intranasal midazolam administration can be decreased by administering lidocaine before intranasal midazolam (preadministered lidocaine) or combining lidocaine with midazolam in a single solution (coadministered lidocaine). We hypothesized coadministered lidocaine is non-inferior to preadministered lidocaine for decreasing pain and distress associated with intranasal midazolam administration. METHODS: Randomized, outcome assessor-blinded, noninferiority trial. Children aged 6 months to 7 years undergoing laceration repair received intranasal midazolam with preadministered or coadministered lidocaine. Pain and distress were evaluated with the Observational Scale of Behavioral Distress-Revised (OSBD-R) (primary outcome; non-inferiority margin 1.8 units) and the Children's Hospital of Eastern Ontario Pain Scale (CHEOPS) and Faces, Legs, Activity, Cry, Consolability (FLACC) scales and cry duration (secondary outcomes). Secondary outcomes also included adverse events, clinician and caregiver satisfaction, and pain and distress associated with intranasal lidocaine administration. RESULTS: Fifty-one patients were analyzed. Mean OSBD-R scores associated with intranasal midazolam administration were 6.4 (95% confidence interval [CI] 5, 7.8) and 7 (95% CI 5.2, 8.9) units for preadministered and coadministered lidocaine, respectively. The difference of 0.6 (95% CI -1.7, 2.8) units represented an inconclusive non-inferiority determination. CHEOPS and FLACC scores and cry duration were similar between groups. OSBD-R, CHEOPS, and FLACC scores and cry duration associated with intranasal lidocaine administration were 3.8, 9.9, and 6 units, and 56 seconds, respectively. Clinicians considered coadministered lidocaine easier to administer. CONCLUSION: Pain and distress associated with intranasal midazolam administration were similar when using coadministered or preadministered lidocaine, but our non-inferiority determination was inconclusive. Administration of intranasal lidocaine by itself was associated with a measurable degree of pain and distress.Keywords: intranasal, midazolam, anxiolysis, sedation, emergency department, emergency medicine, pain, distress, pediatric, lidocaine, laceration.

Caso Clínico: Tiroides Lingual Ectópica en Paciente Adulta Hipotiroidea. Diagnóstico Casual por Tomografía Computarizada. / Clinical Case: Ectopic Lingual Thyroid in Adult Hypothyroid Patient. Casual Diagnosis by Computed Tomography.

Introducción: Los casos de tiroides ectópica localizados en la base de la lengua son anormalidades congénitas raras y difíciles de diagnosticar. Razón de presentación del caso. Caso Clínico: El caso corresponde a una mujer de 41 años con tiroides en base de la lengua diagnosticada incidentalmente con tomografía computarizada (TC), con antecedentes de hipotiroidismo y cáncer de mama derecha. Al examen físico de cuello no se palpa glándula tiroidea ni se observa masa o protuberancia en cavidad bucal. Por control del cáncer de mama, se solicita tomografía por emisión de positrones (PET) y ecografía de cuello, reportándose captación del radiofármaco en la región cervical anterior y superior de cuello, y ausencia de tejido glandular tiroideo a nivel habitual, respectivamente. Por cuanto, se realiza TC simple y contrastada observándose a nivel de la raíz de la lengua una imagen nodular hipercaptante que mide 23x20x20 mm, bien definida, contornos regulares, no infiltra tejidos adyacentes, impronta luz de la orofaringe, sin individualizar la glándula tiroides a nivel habitual, corroborando así el diagnóstico de tiroides ectópica lingual. Conclusión: El diagnóstico de tiroides ectópica en paciente adulto hipotiroideo es raro, por lo que debe considerarse la realización de TC si al examen físico y ecográfico no es palpable ni observable

Introduction: Cases of ectopic thyroid located at the base of the tongue are rare and difficult to diagnose congenital abnormalities. Reason for presenting the case. Clinical case: The case corresponds to a 41-year-old woman with thyroid based on the tongue diagnosed incidentally with computed tomography (CT), with a history of hypothyroidism and right breast cancer. On the physical examination of the neck, the thyroid gland is not palpated, and no mass or bump is observed in the oral cavity. For breast cancer control, positron emission tomography (PET) and neck ultrasound are requested, radiopharmaceutical uptake is reported in the anterior and upper cervical neck region, and absence of thyroid glandular tissue at the usual level, respectively. As a simple and contrasted CT scan, a hypercapting nodular image measuring 23x20x20 mm, well defined, regular contours is observed at the root of the tongue, does not infiltrate adjacent tissues, oropharynx light imprint, without individualizing the thyroid gland to usual level, thus corroborating the diagnosis of lingual ectopic thyroid. Conclusion: The diagnosis of ectopic thyroid in an adult hypothyroid patient is rare, so CT should be considered if the physical and ultrasound examination is not palpable or observable.

High-resolution melting of the cytochrome B gene in fecal DNA: A powerful approach for fox species identification of the Lycalopex genus in Chile.

Easy, economic, precise species authentication is currently necessary in many areas of research and diagnosis in molecular biology applied to conservation studies of endangered species. Here, we present a new method for the identification of three fox species of the Lycalopex genus in Chile. We developed an assay based on high-resolution melt analysis of the mitochondrial cytochrome B gene, allowing a simple, low cost, fast, and accurate species determination. To validate the assay applicability for noninvasive samples, we collected fecal samples in the Atacama Desert, finding unexpectedly one species outside of its known distribution range. We conclude that the assay has a potential to become a valuable tool for a standardized genetic monitoring of the Lycalopex species in Chile.

Dual RNA-seq identifies human mucosal immunity protein Mucin-13 as a hallmark of Plasmodium exoerythrocytic infection.

The exoerythrocytic stage of Plasmodium infection is a critical window for prophylactic intervention. Using genome-wide dual RNA sequencing of flow-sorted infected and uninfected hepatoma cells we show that the human mucosal immunity gene, mucin-13 (MUC13), is strongly upregulated during Plasmodium exoerythrocytic hepatic-stage infection. We confirm MUC13 transcript increases in hepatoma cell lines and primary hepatocytes. In immunofluorescence assays, host MUC13 protein expression distinguishes infected cells from adjacent uninfected cells and shows similar colocalization with parasite biomarkers such as UIS4 and HSP70. We further show that localization patterns are species independent, marking both P. berghei and P. vivax infected cells, and that MUC13 can be used to identify compounds that inhibit parasite replication in hepatocytes. This data provides insights into host-parasite interactions in Plasmodium infection, and demonstrates that a component of host mucosal immunity is reprogrammed during the progression of infection.

Developing Plasmodium vivax Resources for Liver Stage Study in the Peruvian Amazon Region.

To develop new drugs and vaccines for malaria elimination, it will be necessary to discover biological interventions, including small molecules that act against Plasmodium vivax exoerythrocytic forms. However, a robust in vitro culture system for P. vivax is still lacking. Thus, to study exoerythrocytic forms, researchers must have simultaneous access to fresh, temperature-controlled patient blood samples, as well as an anopheline mosquito colony. In addition, researchers must rely on native mosquito species to avoid introducing a potentially dangerous invasive species into a malaria-endemic region. Here, we report an in vitro culture system carried out on site in a malaria-endemic region for liver stage parasites of P. vivax sporozoites obtained from An. darlingi, the main malaria vector in the Americas. P. vivax sporozoites were obtained by dissection of salivary glands from infected An. darlingi mosquitoes and purified by Accudenz density gradient centrifugation. HC04 liver cells were exposed to P. vivax sporozoites and cultured up to 9 days. To overcome low P. vivax patient parasitemias, potentially lower mosquito vectorial capacity, and humid, nonsterile environmental conditions, a new antibiotic cocktail was included in tissue culture to prevent contamination. Culturing conditions supported exoerythrocytic (EEF) P. vivax liver stage growth up to 9 days and allowed for maturation into intrahepatocyte merosomes. Some of the identified small forms were resistant to atovaquone (1 µM) but sensitive to the phosphatidylinositol 4-kinase inhibitor, KDU691 (1 µM). This study reports a field-accessible EEF production process for drug discovery in a malaria-endemic site in which viable P. vivax sporozoites are used for drug studies using hepatocyte infection. Our data demonstrate that the development of meaningful, field-based resources for P. vivax liver stage drug screening and liver stage human malaria experimentation in the Amazon region is feasible.