Dietary heme iron is an important nutritional source of iron in carnivores and omnivores that is more readily absorbed than non-heme iron derived from vegetables and grain. Most heme is absorbed in ...the proximal intestine, with absorptive capacity decreasing distally. We utilized a subtractive hybridization approach to isolate a heme transporter from duodenum by taking advantage of the intestinal gradient for heme absorption. Here we show a membrane protein named HCP1 (heme carrier protein 1), with homology to bacterial metal-tetracycline transporters, mediates heme uptake by cells in a temperature-dependent and saturable manner. HCP1 mRNA was highly expressed in duodenum and regulated by hypoxia. HCP1 protein was iron regulated and localized to the brush-border membrane of duodenal enterocytes in iron deficiency. Our data indicate that HCP1 is the long-sought intestinal heme transporter.
B cell development is a highly regulated process involving multiple differentiation steps, yet many details regarding this pathway remain unknown. Sequencing of patients with B cell-restricted ...immunodeficiency reveals autosomal dominant mutations in TOP2B. TOP2B encodes a type II topoisomerase, an essential gene required to alleviate topological stress during DNA replication and gene transcription, with no previously known role in B cell development. We use Saccharomyces cerevisiae, and knockin and knockout murine models, to demonstrate that patient mutations in TOP2B have a dominant negative effect on enzyme function, resulting in defective proliferation, survival of B-2 cells, causing a block in B cell development, and impair humoral function in response to immunization.
Significance Identifying molecules that are specific to tumors for use in early detection, diagnosis, prognosis, and therapy is both a primary goal and a key discovery challenge across diverse areas ...of oncology. To discover ovarian tumor-specific molecules, we developed custom bioinformatics algorithms to analyze transcriptome sequence data of 296 ovarian cancer and 1,839 normal tissues and validated putative tumor-specific mRNA isoforms by RT–quantitative PCR. The results revealed multiple candidate diagnostic and therapeutic targets with unique sequences that were expressed in most of the cancers examined but not in normal tissues. The process we developed can be readily applied to identify diagnostic and therapeutic targets for any of the 30 or more tumor types for which large amounts of transcriptome data now exist.
Tumor-specific molecules are needed across diverse areas of oncology for use in early detection, diagnosis, prognosis and therapy. Large and growing public databases of transcriptome sequencing data (RNA-seq) derived from tumors and normal tissues hold the potential of yielding tumor-specific molecules, but because the data are new they have not been fully explored for this purpose. We have developed custom bioinformatic algorithms and used them with 296 high-grade serous ovarian (HGS-OvCa) tumor and 1,839 normal RNA-seq datasets to identify mRNA isoforms with tumor-specific expression. We rank prioritized isoforms by likelihood of being expressed in HGS-OvCa tumors and not in normal tissues and analyzed 671 top-ranked isoforms by high-throughput RT-qPCR. Six of these isoforms were expressed in a majority of the 12 tumors examined but not in 18 normal tissues. An additional 11 were expressed in most tumors and only one normal tissue, which in most cases was fallopian or colon. Of the 671 isoforms, the topmost 5% ( n = 33) ranked based on having tumor-specific or highly restricted normal tissue expression by RT-qPCR analysis are enriched for oncogenic, stem cell/cancer stem cell, and early development loci—including ETV4, FOXM1, LSR, CD9, RAB11FIP4, and FGFRL1. Many of the 33 isoforms are predicted to encode proteins with unique amino acid sequences, which would allow them to be specifically targeted for one or more therapeutic strategies—including monoclonal antibodies and T-cell–based vaccines. The systematic process described herein is readily and rapidly applicable to the more than 30 additional tumor types for which sufficient amounts of RNA-seq already exist.
Human papilloma virus-16 (HPV-16) infection is a major risk factor for a subset of head and neck squamous cell carcinoma (HNSCC), in particular oropharyngeal squamous cell carcinoma (OPSCC). Current ...techniques for assessing the HPV-16 status in HNSCC include the detection of HPV-16 DNA and p16(INK4a) expression in tumor tissues. When tumors originate from hidden anatomical sites, this method can be challenging. A non-invasive and cost-effective alternative to biopsy is therefore desirable for HPV-16 detection especially within a community setting to screen at-risk individuals.
The present study compared detection of HPV-16 DNA and RNA in salivary oral rinses with tumor p16(INK4a) status, in 82 HNSCC patients using end-point and quantitative polymerase chain reaction (PCR).
Of 42 patients with p16(INK4a)-positive tumours, 39 (sensitivity = 92.9 %, PPV = 100 % and NPV = 93 %) had oral rinse samples with detectable HPV-16 DNA, using end-point and quantitative PCR. No HPV-16 DNA was detected in oral rinse samples from 40 patients with p16(INK4a) negative tumours, yielding a test specificity of 100 %. For patients with p16(INK4a) positive tumours, HPV-16 mRNA was detected using end-point reverse transcription PCR (RT-PCR) in 24/40 (sensitivity = 60 %, PPV = 100 % and NPV = 71 %), and using quantitative RT-PCR in 22/40 (sensitivity = 55 %, PPV = 100 % and NPV = 69 %). No HPV-16 mRNA was detected in oral rinse samples from the p16(INK4a)-negative patients, yielding a specificity of 100 %.
We demonstrate that the detection of HPV-16 DNA in salivary oral rinse is indicative of HPV status in HNSCC patients and can potentially be used as a diagnostic tool in addition to the current methods.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Vaccines delivered to the skin by microneedles—with and without adjuvants—have increased immunogenicity with lower doses than standard vaccine delivery techniques such as intramuscular or intradermal ...injection. However, the mechanisms underlying this skin-mediated “adjuvant” effect are not clear. Here, we show that the dynamic application of a microprojection array (the Nanopatch) to skin generates localized transient stresses invoking cell death around each projection. Nanopatch application caused significantly higher levels (∼65-fold) of cell death in murine ear skin than i.d. injection using a hypodermic needle. Measured skin cell death is associated with modeled stresses ∼1–10MPa. Nanopatch-immunized groups also yielded consistently higher anti-immunoglobulin G endpoint titers (up to 50-fold higher) than i.d. groups after delivery of a split virion influenza vaccine. Importantly, colocalization of cell death with nearby live skin cells and delivered antigen was necessary for immunogenicity enhancement. These results suggest a correlation between cell death caused by the Nanopatch with increased immunogenicity. We propose that the localized cell death serves as a “physical immune enhancer” for the adjacent viable skin cells, which also receive antigen from the projections. This natural immune enhancer effect has the potential to mitigate or replace chemical-based adjuvants in vaccines.
Background Human papilloma virus-16 (HPV-16) infection is a major risk factor for a subset of head and neck squamous cell carcinoma (HNSCC), in particular oropharyngeal squamous cell carcinoma ...(OPSCC). Current techniques for assessing the HPV-16 status in HNSCC include the detection of HPV-16 DNA and p16.sup.INK4a expression in tumor tissues. When tumors originate from hidden anatomical sites, this method can be challenging. A non-invasive and cost-effective alternative to biopsy is therefore desirable for HPV-16 detection especially within a community setting to screen at-risk individuals. Methods The present study compared detection of HPV-16 DNA and RNA in salivary oral rinses with tumor p16.sup.INK4a status, in 82 HNSCC patients using end-point and quantitative polymerase chain reaction (PCR). Results Of 42 patients with p16.sup.INK4a-positive tumours, 39 (sensitivity = 92.9 %, PPV = 100 % and NPV = 93 %) had oral rinse samples with detectable HPV-16 DNA, using end-point and quantitative PCR. No HPV-16 DNA was detected in oral rinse samples from 40 patients with p16.sup.INK4a negative tumours, yielding a test specificity of 100 %. For patients with p16.sup.INK4a positive tumours, HPV-16 mRNA was detected using end-point reverse transcription PCR (RT-PCR) in 24/40 (sensitivity = 60 %, PPV = 100 % and NPV = 71 %), and using quantitative RT-PCR in 22/40 (sensitivity = 55 %, PPV = 100 % and NPV = 69 %). No HPV-16 mRNA was detected in oral rinse samples from the p16.sup.INK4a-negative patients, yielding a specificity of 100 %. Conclusions We demonstrate that the detection of HPV-16 DNA in salivary oral rinse is indicative of HPV status in HNSCC patients and can potentially be used as a diagnostic tool in addition to the current methods. Keywords: HPV, HNSCC, OPSCC, Saliva, Early detection
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The presence of short-lived radioisotopes (SLRs) in solar system meteorites has been interpreted as evidence that the solar system was exposed to a supernova shortly before or during its formation. ...Yet results from hydrodynamical models of SLR injection into the proto-solar cloud or disc suggest that gas-phase mixing may not be efficient enough to reproduce the observed abundances. As an alternative, we explore the injection of SLRs via dust grains as a way to overcome the mixing barrier. We numerically model the interaction of a supernova remnant containing SLR-rich dust grains with a nearby molecular cloud. The dust grains are subject to drag forces and both thermal and non-thermal sputtering. We confirm that the expanding gas shell stalls upon impact with the dense cloud and that gas-phase SLR injection occurs slowly due to hydrodynamical instabilities at the cloud surface. In contrast, dust grains of sufficient size (> 1 micron) decouple from the gas and penetrate into the cloud within 0.1 Myr. Once inside the cloud, the dust grains are destroyed by sputtering, releasing SLRs and rapidly enriching the dense (potentially star-forming) regions. Our results suggest that SLR transport on dust grains is a viable mechanism to explain SLR enrichment.
Our aim was to develop a widely available educational program in which students conducted authentic research that met the expectations of both the scientific and educational communities. This paper ...describes the development and implementation of a citizen science project based on DNA barcoding of reptile specimens obtained from the Museums Victoria frozen tissue collection. The student program was run by the Gene Technology Access Centre (GTAC) and was delivered as a "one day plus one lesson" format incorporating a one-day wet laboratory workshop followed by a single lesson at school utilising online bioinformatics tools. The project leveraged the complementary resources and expertise of the research and educational partners to generate robust scientific data that could be analysed with confidence, meet the requirements of the Victorian state education curriculum, and provide participating students with an enhanced learning experience. During two 1-week stints in 2013 and 2014, 406 students mentored by 44 postgraduate university students participated in the project. Students worked mainly in pairs to process ~200 tissue samples cut from 53 curated reptile specimens representing 17 species. A total of 27 novel Cytochrome Oxidase subunit 1 (CO1) sequences were ultimately generated for 8 south-east Australian reptile species of the families Scincidae and Agamidae.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Inner phenomena, such as personal motivations for pursuing sustainability, may be critical levers for improving conservation outcomes. Most conservation research and policies, however, focus on ...external phenomena (e.g., ecological change or economic processes). We explored the factors shaping 9 conservation attitudes toward forest and wildlife protection among colonist farmers around an Amazonian deforestation frontier. Our data comprised 241 face‐to‐face quantitative surveys, complemented with qualitative insights from open‐ended questionnaire responses and opportunistic semistructured interviews. To account for the full spectrum of possible inner motivations, we employed measures of nature connection (indicating biospheric motivation) and personal values organized around the traditionalism (traditionalist through to high openness to change) and universalism dimensions (egoistic through to altruistic motivations). We used averaged beta‐binomial generalized linear models to assess the role of external factors (socioeconomic, sociodemographic, and environmental) and personal (inner) motivations on the variation in attitudes. Each attitude was modeled separately. The relative importance of each predictor was judged by the proportion of models where it appeared as significant. Proconservation views were expressed by the majority (at least 65%) of the respondents in 7 out of the 9 attitude models. The most consistent predictors were emotional nature connection and personal values (significant in 4–6 out of 9 models), rather than external phenomena (significant in 0–5 models). However, the poorest farmers had lower scores on the agreement with prioritizing nature over development (𝛽 = –0.52, 95% CI: –0.96 to –0.07). Qualitative data also indicated that economic barriers hinder forest conservation on farms. These results suggest that biospheric, traditionalistic, and altruistic motivations promote people's proconservation attitudes, but nurturing these latent motivations is unlikely to improve conservation outcomes if material poverty remains unaddressed. Integrating the inner–outer perspective into conservation thinking and practical interventions could foster environmental stewardship and increase human well‐being.
Evaluación de la influencia de la conexión y los valores naturales sobre las actitudes de conservación es una frontera tropical de deforestación
Resumen
Los fenómenos interiores, como los motivos personales para ser sustentables, pueden ser niveladores importantes para aumentar los resultados de conservación. Sin embargo, la mayor parte de la investigación y las políticas se enfocan en los fenómenos exteriores (cambios ecológicos o procesos económicos, por ejemplo). Exploramos los factores que moldean nueve actitudes de conservación relacionadas con la protección de los bosques y la fauna en un grupo de agricultores colonos cercanos a una frontera de deforestación en la Amazonía. Nuestros datos se obtuvieron de 241 encuestas presenciales cuantitativas complementadas con información cualitativa tomada de las respuestas en cuestionarios de preguntas abiertas y entrevistas oportunistas semiestructuradas. Para considerar el espectro completo de las posibles motivaciones interiores empleamos medidas de la conexión natural (lo que indica motivos relacionados a la biósfera) y valores personales organizados en torno a las dimensiones del tradicionalismo (del tradicionalista hasta una gran apertura al cambio) y el universalismo (del egoísta hasta los motivos altruistas). Usamos modelos lineales generalizados betabinomiales promediados para evaluar el papel que tienen los factores externos (socioeconómicos, sociodemográficos y ambientales) y los motivos personales (interiores) en la variación de las actitudes. Modelamos cada actitud por separado. Juzgamos la importancia relativa de cada predictor mediante la proporción de modelos en los que aparecían como significativos. La mayoría (al menos el 65%) de los respondientes expresó las opiniones en pro de la conservación en siete de los nueve modelos de actitud. Los predictores más uniformes fueron la conexión emocional natural y los valores personales (significativos en cuatro a seis de los nueve modelos), en lugar de los fenómenos externos (significativos en cero a cinco modelos). Sin embargo, los agricultores más pobres tuvieron un puntaje más bajo en cuanto a estar de acuerdo con la priorización de la naturaleza sobre el desarrollo (𝛽 = ‐0.52, 95% CI ‐0.96 a ‐0.07). Los datos cualitativos también indicaron que las barreras económicas impiden la conservación de los bosques en las fincas. Estos resultados sugieren que los motivos tradicionalistas, altruistas y aquellos relacionados con la biósfera promueven las actitudes en pro de la conservación de las personas, pero es poco probable que propiciar estos motivos latentes aumente los resultados de conservación si sigue sin solucionarse la pobreza material. La inclusión de las perspectivas internas y externas dentro del pensamiento de conservación y las intervenciones prácticas podría fomentar la administración ambiental e incrementar el bienestar humano.
【摘要】
内在现象, 如追求可持续性的个人动机, 可能是改善保护结果的关键杠杆。然而, 大多数保护研究和政策都仅关注外在现象, 如生态变化或经济过程。本研究探讨了在亚马逊森林砍伐前线的殖民农民中, 影响对森林和野生动物保护的 9 种保护态度的因素。我们的数据包含 241 个面对面的定量调查, 辅以来自开放式问卷调查和机会性半结构化访谈的定性调查结果。为了说明所有可能的内在动机, 我们采用了对自然联系 (生物圈动机) 和个人价值观的测量, 围绕传统主义 (从传统主义者到对变化高度接受) 和普遍主义维度 (从利己主义动机到利他主义动机) 。我们使用了平均 beta 二项式广义线性模型评估外在因素 (社会经济、社会人口和环境) 和个人 (内在) 动机在态度变化中的作用, 并对每种态度单独建模, 并用每个预测因子在模型中出现的显著比例来确定其相对重要性。结果表明, 在 9 个态度模型的 7 个中, 大多数 (至少 65%) 受访者表达了支持保护的观点。最一致的预测因子是与自然的情感联系和个人价值观 (在9个模型的 4–6 个模型中显著), 而不是外在现象 (在 0–5 个模型中显著) 。然而, 最贫穷的农民在认同自然优先于发展方面的打分较低 (𝛽= −0.52, 95% 置信区间为 −0.96 至 −0.07) 。 定性数据还表明, 经济障碍阻碍了农场的森林保护。以上结果表明, 生物圈、传统主义和利他主义动机促进了人们支持保护的态度, 但如果物质贫困问题得不到解决, 培养这些潜在动机也不太可能改善保护结果。我们提出, 将内在‐外在视角纳入保护思想和实际干预措施, 有助于促进环境管理并提高人类福祉。【翻译: 胡怡思; 审校: 聂永刚】
Drug repurposing is promoted as a cost- and time-effective mechanism for providing new medicines. Often, however, there is insufficient consideration by academic researchers of the processes required ...to ensure that a repurposed drug can be used for a new indication. This may explain the inability of drug repurposing to fulfill its promise. Important aspects, often overlooked, include financial and intellectual property considerations, the clinical and regulatory path, and clinical equipoise, which provides ethical justification for randomized controlled trials. The goal of drug repurposing is to obtain a new regulator-approved label for an existing drug, and so, the trajectory for drug repurposing and traditional drug development is similar. Here, we discuss factors critical for a successful repurposed medicine to help academic investigators better identify drug repurposing opportunities.
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