bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma membrane cholesterol trafficking in steroidogenesis
Authors: Deng Bing , Shen Wen-Jun , Dong Dachuan , Azhar Salman , Kraemer Fredric B.
Published: August 2018

Cholesterol is an important component of plasma membranes (PMs) and the precursor of all steroid hormones. In steroidogenic tissues, upon hormone stimulation, there is a rapid transfer of cholesterol to the mitochondria, which is the site of the initial step in steroidogenesis. In the current study, we examined PM cholesterol trafficking for steroidogenesis. In a mitochondrial reconstitution assay, adrenal PMs supported steroidogenesis in the absence of additional transport proteins. Depletion of cholesterol in PMs by 50% eliminated the membranes' ability to support steroidogenesis in vitro and reduced steroid production in intact Y1 adrenocortical cells. Syntaxin (STX)‐5 and α‐soluble N‐emylmaleimide‐sensitive factor attachment protein (α‐SNAP) are enriched in adrenal PMs, and adrenocorticotropic hormone treatment of rats recruited STX5 and α‐SNAP to adrenal PMs and mitochondria. Immunodepletion of STX5 and α‐SNAP from PMs decreased steroidogenesis supported by PMs in vitro. Protease digestion of PMs decreased, whereas recombinant STX5 or α‐SNAP restored, the PMs' ability to support steroidogenesis. Knockdown of either STX5 or α‐SNAP in Y1 cells decreased stimulated steroidogenesis. These results indicate that STX5 and α‐SNAP facilitate cholesterol trafficking from PMs to mitochondria for adrenal steroid synthesis and underscore the importance of vesicular trafficking of PM cholesterol for steroidogenesis.—Deng, B., Shen, W.‐J., Dong, D., Azhar, S., Kraemer, F. B. Plasma membrane cholesterol trafficking in steroidogenesis. FASEB J. 33, 389–1400 (2019). www.fasebj.org
Chronic psychosocial defeat differently affects lipid metabolism in liver and white adipose tissue and induces hepatic oxidative stress in mice fed a high‐fat diet
Authors: Giudetti Anna M. , Testini Mariangela , Vergara Daniele , Priore Paola , Damiano Fabrizio , Gallelli Cristina Anna , Romano Adele , Villani Rosanna , Cassano Tommaso , Siculella Luisa , Gnoni Gabriele V. , Moles Anna , Coccurello Roberto , Gaetani Silvana
Published: August 2018

It is widely accepted that chronic stress may alter the homeostatic mechanisms of body weight control. In this study, we followed the metabolic changes occurring in mice when chronic stress caused by psychosocial defeat (CPD) is associated with ad libitum exposure to a palatable high‐fat diet (HFD). In this model, CPD mice consumed more HFD than unstressed (Un) mice without gaining body weight. We focused on metabolic processes involved in weight control, such as de novo lipogenesis (DNL), fatty acid β‐oxidation (FAO), and thermogenesis. The activity and expression of DNL enzymes were reduced in the liver and white adipose tissue of mice consuming the HFD. Such effects were particularly evident in stressed mice. In both CPD and Un mice, HFD consumption increased the hepatic expression of the mitochondrial FAO enzyme carnitine palmitoyltransferase‐1. In the liver of mice consuming the HFD, stress exposure prevented accumulation of triacylglycerols; however, accumulation of triacylglycerols was observed in Un mice under the same dietary regimen. In brown adipose tissue, stress increased the expression of uncoupling protein‐1, which is involved in energy dissipation, both in HFD and control diet‐fed mice. We consider increased FAO and energy dissipation responsible for the antiobesity effect seen in CPD/HFD mice. However, CPD associated with HFD induced hepatic oxidative stress.—Giudetti, A. M., Testini, M., Vergara, D., Priore, P., Damiano, F., Gallelli, C. A., Romano, A., Villani, R., Cassano, T., Siculella, L., Gnoni, G. V., Moles, A., Coccurello, R., Gaetani, S. Chronic psychosocial defeat differently affects lipid metabolism in liver and white adipose tissue and induces hepatic oxidative stress in mice fed a high‐fat diet. FASEB J. 33, 1428–1439 (2019). www.fasebj.org
Defining Empathy to Better Teach, Measure, and Understand Its Impact
Authors: Dohrenwend Anne M.
Published: August 2018

No consensus on the definition of empathy exists. Empathy has been described as emotional and spontaneous, cognitive and deliberate, or some combination of the two. Attentive nonverbal reactions, feeling reflections, reassurance, sympathy, and compassion all have been conflated with empathy, making it impossible to differentiate empathy from other communication skills. This confusion over the definition of empathy has affected its measurement. For example, the authors of the Interpersonal Reactivity Index see empathy as multidimensional, involving both emotional and cognitive aspects, while the authors of the Jefferson Scale of Physician Empathy see empathy as a predominately cognitive process. Researchers, such as Yamada and colleagues in their study in this issue, then must straddle these conceptualizations, which is a limitation to their work.

To address this problem, the author of this Invited Commentary proposes adopting the cognitive definition of empathy, noting that it allows physicians to distinguish between empathy and other communication skills and is the most consistent with counseling psychology descriptions. The author defines cognitive empathy as a conscious, strenuous, mental effort to clarify a patient’s muddy expression of her experience using a soft interpretation of her story. Accurate empathic responses are unadulterated by a physician’s reactions to a patient and the patient’s experience. The author describes four aspects of empathy that are rarely noted in the medical literature but that are fundamental to understanding its practice: the “as if” condition, the use of soft interpretation, the primacy of cognition, and the relevance of reflection.
Pilomatrixoma: A Comprehensive Review of the Literature
Authors: Jones Christopher D. , Ho Weiguang , Robertson Bernard F. , Gunn Eilidh , Morley Stephen
Published: August 2018

Introduction:

Pilomatrixoma, also known as calcifying epithelioma of Malherbe, is a superficial benign skin tumor that arises from hair follicle matrix cells. Although pilomatrixomas are well-recognized lesions, clinically they are frequently misdiagnosed as other skin conditions. By reviewing all the literature over the past 10 years, the aims of this article are to analyze the cause, clinical presentation, management, and outcome of pilomatrixoma among children and adults to gain a more complete understanding of this lesion in today's clinical context.

Methods:

A MEDLINE and EMBASE search was conducted from January 2005 to February 2015 using a combination of the terms: “child,” “childhood,” “adult,” and keywords: “pilomatrixoma,” “pilomatricoma,” and “calcifying epithelioma of Malherbe.” A total of 150 articles were reviewed.

Results:

The lesions occurred most commonly in the first and second decades (mean age 16 years and 7 months). The commonest presentation was of an asymptomatic, firm, slowly growing, mobile nodule. Only 16% were accurately diagnosed on clinical examination. Imaging in the form of ultrasound, computed tomography, and magnetic resonance imaging has been reported. Pathological diagnosis was achieved through incision, punch, and shave biopsies. Pathological findings are discussed and summarized in this review.

Conclusion:

Pilomatrixomas are thought to arise from mutation in the Wnt pathway and has been linked to several genetic conditions. It is commonly misdiagnosed preoperatively; however, with better awareness of the lesion, it can be appropriately treated while avoiding unnecessary diagnostic tests. Complete surgical excision with clear margins is almost always curative.
Do Infant Feeding Practices Differ Between Grandmothers and Mothers in Rural China? Evidence From Rural Shaanxi Province
Authors: Yue Ai , Zhang Nianrui , Liu Xueyang , Tang Lei , Luo Renfu , Yang Meredith , Rozelle Scott , Medina Alexis
Published: August 2018

The overall goal of this study is to examine whether infant feeding practices differ between mothers and grandmothers in rural China. We randomly sampled 1383 caregivers of infants aged 18 to 30 months living in 351 villages across 174 townships in nationally designated poverty counties in rural areas. Results show that a high fraction of caregivers of 18- to 30-month-old children living in low-income areas of rural China do not regularly engage in positive infant feeding practices. Only 30% of children in our sample achieved adequate dietary diversity. Only 49% of children in our sample were fed meat in the day prior to survey administration. Few caregivers reported giving any vitamin supplements (such as calcium or iron supplements) to their children. We find that 33% of the children were cared for by grandmothers rather than mothers, and that grandmothers feed a less diversified diet to children than do mothers. Most (84%) caregivers rely solely on their own experiences, friends, and family members in shaping their feeding behaviors. Overall infant feeding practices are poor in rural China. Grandmothers engage in poorer feeding practices than do mothers. Grandmothers have improved their feeding practices compared to when their own children were young. Our results suggest shortcomings in the quality of infant feeding practices, at least in part due to an absence of reliable information sources.
Embryologic and Fetal Development of the Human Orbit
Authors: Tawfik Hatem A. , Dutton Jonathan J.
Published: August 2018

Purpose:

To review the recent data about orbital development and sort out the controversies from the very early stages during embryonic life till final maturation of the orbit late in fetal life, and to appreciate the morphogenesis of all the definitive structures in the orbit in a methodical and timely fashion.

Methods:

The authors extensively review major studies detailing every aspect of human embryologic and fetal orbital morphogenesis including the development of extraocular muscles, orbital fat, vessels, nerves, and the supportive connective tissue framework as well as bone. These interdisciplinary studies span almost a century and a half, and include some significant controversial opposing points of view which the authors hopefully sort out. The authors also highlight a few of the most noteworthy molecular biologic studies regarding the multiple and interacting signaling pathways involved in regulating normal orbital morphogenesis.

Results:

Orbital morphogenesis involves a successive series of subtle yet tightly regulated morphogenetic events that could only be explained through the chronological narrative used by the authors. The processes that trigger and contribute to the formation of the orbits are complex and seem to be intricately regulated by multifaceted interactions and bidirectional cross-talk between a multitude of cellular building raw materials including the developing optic vesicles, neuroectoderm, cranial neural crest cells and mesoderm.

Conclusions:

Development of the orbit is a collective enterprise necessitating interactions between, as well as contributions from different cell populations both within and beyond the realm of the orbit. A basic understanding of the processes underlying orbital ontogenesis is a crucial first step toward establishing a genetic basis or an embryologic link with orbital disease.
Efficacy of Care for Blind Painful Eyes
Authors: Idowu Oluwatobi O. , Ashraf Davin C. , Kalin-Hajdu Evan , Ryan Michael C. , Kersten Robert C. , Vagefi M. Reza
Published: August 2018

Purpose:

Pain relief for a blind painful eye often follows an escalating paradigm of interventions. This study compares the efficacy of common interventions.

Methods:

A retrospective chart review of blind painful eye cases was conducted at a single tertiary institution from April 2012 to December 2016. Demographics, etiology, treatment, and pain level were assessed.

Results:

Among 99 blind painful eyes, 96 eyes initially received medical therapy (topical steroids, cycloplegics, and/or hypotensives), with pain relief in 39% of eyes. Minimally invasive interventions (laser cyclophotocoagulation, retrobulbar injection, or corneal electrocautery) were performed 41 times in 36 eyes, 34 of which had failed medical therapy, and led to pain relief in 75% of eyes. Evisceration or enucleation was performed in 28 eyes, and long-term pain relief was achieved in 100% of eyes. Surgery allowed discontinuation of oral analgesics in 100% of cases versus 20% for minimally invasive therapy (p = 0.005) and 14% for medical therapy (p = 0.0001). Compared with medical therapy, minimally invasive therapy was 2.5 times more likely to achieve lasting pain relief (p = 0.003) and surgical therapy 35.6 times more likely to achieve lasting pain relief (p = 0.011). High initial pain score was associated with nonsurgical treatment failure.

Conclusions:

Medical therapy provides pain relief in a moderate number of patients with a blind painful eye. When medical therapy fails, minimally invasive therapy and surgical interventions are successively more effective in relieving ocular pain. High initial pain score is a risk factor for nonsurgical therapy failure and may merit an earlier discussion of surgical intervention.
Barriers to Medication Access in Pediatric Oncology in the United States
Authors: Kelly Michael J. , Sung Lillian , Dickens David S.
Published: August 2018

Timely medication access in pediatric oncology is important; yet barriers are poorly described. We surveyed pediatric oncology health care providers at National Cancer Institute Community Oncology Research Program sites on their experience with the impact of drug acquisition difficulties, prior authorization (PA) requests, insurance denials, and patient copays leading to deviations or delays from prescribed treatment for their pediatric/adolescent/young adult patients in calendar year 2016. PA requests, the most frequently cited issue, created a deviation or delay from planned chemotherapy and supportive care treatment in at least 61% of respondents. Half of the respondents believed that PA-induced delays had a negative impact on care. Two-thirds of respondents felt that delays in starting therapy due to barriers in medication access created psychologic or emotional distress for the patient or family. Pediatric cancer patients may be receiving inferior care as a result of barriers to medication access.
Immune responses to dengue virus in the skin
Authors: Rathore Abhay P. S. , St. John Ashley L.
Published: August 2018

Dengue virus (DENV) causes infection in humans and current estimates place 40% of the world population at risk for contracting disease. There are four DENV serotypes that induce a febrile illness, which can develop into a severe and life-threatening disease in some cases, characterized primarily by vascular dysregulation. As a mosquito-borne infection, the skin is the initial site of DENV inoculation and also where primary host immune responses are initiated. This review discusses the early immune response to DENV in the skin by both infection target cells such as dendritic cells and by immune sentinels such as mast cells. We provide an overview of the mechanisms of immune sensing and functional immune responses that have been shown to aid clearance of DENV
in vivo
. Finally, we discuss factors that can influence the immune response to DENV in the skin, such as mosquito saliva, which is co-injected with virus during natural route infection, and pre-existing immunity to other DENV serotypes or to related flaviviruses.
Identification and quantification of spinochromes in body compartments of
Echinometra mathaei
’s coloured types
Authors: Brasseur Lola , Demeyer Marie , Decroo Corentin , Caulier Guillaume , Flammang Patrick , Gerbaux Pascal , Eeckhaut Igor
Published: August 2018

Sea urchin pigmentation is mainly due to polyhydroxy-1,4-naphthoquinones called spinochromes. If their molecular structures are well known in test and spines of many species, their abundance and distribution in other body compartments remain unstudied. The aim of this study is to analyse the pigment composition in four body compartments (test/spines, digestive system, gonads and coelomic fluid) of four coloured types of the sea urchin
Echinometra mathaei
. Qualitative and quantitative measurements by mass spectrometry highlight the existence of 13 different pigments; among which are five isomers of known spinochromes as well as three potentially new ones. The composition comparison shows the largest spinochrome diversity in ‘test/spines’ body compartments. The spinochrome concentrations vary from 48 to 1279 mg kg
−1
of dried body compartment. It is the highest in the digestive system, although it is also important in the organic fraction of the ‘test/spines’ body compartment. This observation may be explained by higher exposures of some body compartments to external environments and by the protective role fulfilled by spinochromes against microorganisms, ultraviolet radiation and reactive oxygen species. The ‘black’ type—the most common coloured type in coral reefs—has the highest concentration of spinochromes indicating their importance in Echinoids' fitness by acting as a protective agent.
tarsal-lessis expressed as a gap gene but has no gap gene phenotype in the moth midgeClogmia albipunctata
Authors: Jiménez-Guri Eva , Wotton Karl R. , Jaeger Johannes
Published: August 2018

Gap genes are involved in segment determination during early development of the vinegar flyDrosophila melanogasterand other dipteran insects (flies, midges and mosquitoes). They are expressed in overlapping domains along the antero-posterior (A–P) axis of the blastoderm embryo. While gap domains cover the entire length of the A–P axis inDrosophila,there is a region in the blastoderm of the moth midgeClogmia albipunctata, which lacks canonical gap gene expression. Is a non-canonical gap gene functioning in this area? Here, we characterizetarsal-less(tal) inC. albipunctata. The homologue oftalin the flour beetleTribolium castaneum(calledmilles-pattes, mlpt) is abona fidegap gene. We find thatCa-talis expressed in the region previously reported as lacking gap gene expression. Using RNA interference, we study the interaction ofCa-talwith gap genes. We show thatCa-talis regulated by gap genes, but only has a very subtle effect ontailless (Ca-tll),while not affecting other gap genes at all. Moreover, cuticle phenotypes ofCa-taldepleted embryos do not show any gap phenotype. We conclude thatCa-talis expressed and regulated like a gap gene, but does not function as a gap gene inC. albipunctata.
Habitual stone-tool-aided extractive foraging in white-faced capuchins,Cebus capucinus
Authors: Barrett Brendan J. , Monteza-Moreno Claudio M. , Dogandžić Tamara , Zwyns Nicolas , Ibáñez Alicia , Crofoot Margaret C.
Published: August 2018

Habitual reliance on tool use is a marked behavioural difference between wild robust (genusSapajus) and gracile (genusCebus) capuchin monkeys. Despite being well studied and having a rich repertoire of social and extractive foraging traditions,Cebussp. rarely use tools and have never been observed using stone tools. By contrast, habitual tool use bySapajusis widespread. We review theory and discuss factors which might explain these differences in patterns of tool use betweenCebusandSapajus. We then report the first case of habitual stone tool use in a gracile capuchin: a population of white-faced capuchins (Cebus capucinus imitator) in Coiba National Park, Panama who habitually rely on hammerstone and anvil tool use to access structurally protected food items in coastal areas includingTerminalia catappaseeds, hermit crabs, marine snails, terrestrial crabs and other items. This behaviour has persisted on one island in Coiba National Park since at least 2004. From 1 year of camera trapping, we found that stone tool use is strongly male-biased. Of the 205 camera trap days where tool use was recorded, adult females were never observed to use stone tools, although they were frequently recorded at the sites and engaged in scrounging behaviour. Stone tool use occurs year-round in this population; over half of all identifiable individuals were observed participating. At the most active tool use site, 83.2% of days where capuchins were sighted corresponded with tool use. Capuchins inhabiting the Coiba archipelago are highly terrestrial, under decreased predation pressure and potentially experience resource limitation compared to mainland populations—three conditions considered important for the evolution of stone tool use. White-faced capuchin tool use in Coiba National Park thus offers unique opportunities to explore the ecological drivers and evolutionary underpinnings of stone tool use in a comparative within- and between-species context.
Entrainment and maintenance of an internal metronome in premotor cortex
Authors: Cadena-Valencia J , García-Garibay O , Merchant H , Jazayeri M , de Lafuente V
Published: August 2018

To prepare timely motor actions we constantly predict future events. Regularly repeating events are often perceived as a rhythm to which we can readily synchronize our movements, just as in dancing to music. However, the neuronal mechanisms underlying the capacity to encode and maintain rhythms are not understood. We trained nonhuman primates to maintain the rhythm of a visual metronome of different tempos and then we recorded neural activity in the supplementary motor area (SMA). SMA exhibited rhythmic bursts of gamma band (30-40 Hz) reflecting an internal tempo that matched the extinguished visual metronome. Moreover, gamma amplitude increased throughout the trial and provided an estimate of total elapsed time. Notably, the timing and amplitude of gamma bursts reflected systematic timing biases and errors in the behavioral responses. Our results indicate that premotor areas use dynamic motor plans to encode a metronome for rhythms and a stopwatch for total elapsed time.
Dynamic representations of faces in the human ventral visual stream link visual features to behaviour
Authors: Dima Diana C. , Singh Krish D.
Published: August 2018

Humans can rapidly extract information from faces even in challenging viewing conditions, yet the neural representations supporting this ability are still not well understood. Here, we manipulated the presentation duration of backward-masked facial expressions and used magnetoencephalography (MEG) to investigate the computations underpinning rapid face processing. Multivariate analyses revealed two stages in face perception, with the ventral visual stream encoding facial features prior to facial configuration. When presentation time was reduced, the emergence of sustained featural and configural representations was delayed. Importantly, these representations explained behaviour during an expression recognition task. Together, these results describe the adaptable system linking visual features, brain and behaviour during face perception.
Novel cholera toxin variant and ToxT regulon in environmentalVibrio mimicusstrains: potential resources for the evolution ofVibrio choleraehybrid strains
Authors: Neogi Sucharit Basu , Chowdhury Nityananda , Awasthi Sharda Prasad , Asakura Masahiro , Mahmud Zahid Hayat , Islam Mohammad Sirajul , Hinenoya Atsushi , Nair Gopinath Balakrish , Yamasaki Shinji
Published: August 2018

Atypical El Tor strains ofVibrio choleraeO1 harboring variantctxBgenes of cholera toxin (CT) are gradually becoming a major cause of recent cholera epidemics.Vibrio mimicusoccasionally contains virulence factors associated with cholera, e.g., CT, encoded byctxABon CTXФ genome; and TCP, the CTXФ-specific receptor. This study carried out extensive molecular characterization of CTXФ and ToxT regulon inctx+vestrains ofV. mimicusisolated from the Bengal coast. Southern hybridization, PCR, and DNA sequencing of virulence related-genes revealed the presence of an El Tor type CTX prophage (CTXET) carrying a novelctxAB, tandem copies of environmental type pre-CTX prophage (pre-CTXEnv), and RS1 elements, which were organized in an array of RS1-CTXET-RS1-pre-CTXEnv-pre-CTXEnv. Additionally, a novel variant oftcpAandtoxTrespectively, showing clonal lineage to a phylogenetic clade ofV. choleraenon-O1/O139, was identified. TheV. mimicusstrains lacked the RTX and TLC elements, andVibrioseventh pandemic islands of the El Tor strains, but contained five heptamer (TTTTGAT) repeats inctxABpromoter region like some classical strains ofV. choleraeO1. PFGE analysis showed all thectx+veV. mimicusstrains were clonally related. However, theirin vitroCT production andin vivotoxigenecity were variable, which could be explained by differential transcription of virulence genes along with ToxR regulon. Taken together, our findings strongly suggest that environmentalV. mimicusstrains act as potential reservoir of atypical virulence factors, including variant CT and ToxT regulon, and may contribute to the evolution ofV. choleraehybrid strains.

IMPORTANCE

Natural diversification of CTXФ andctxABgenes certainly influences disease severity and shifting patterns in major etiological agents of cholera, e.g., the overwhelming emergence of hybrid El Tor variants, replacing the prototype El Tor strains ofV. cholerae. This study showing the occurrence of CTXETcomprising a novel variant ofctxABinV. mimicuspoints out a previously unnoticed evolutionary event, independent to that of the El Tor strains ofV. cholerae. Identification and cluster analysis of the newly-discovered alleles oftcpAandtoxTindicates their horizontal transfer from an uncommon clone ofV. cholerae. The genomic content of ToxT regulon, and tandemly arranged multiple pre-CTXФEnvand a CTXФETinV. mimicusprobably act as salient raw materials inducing natural recombination among the hallmark virulence genes of hybridV. choleraestrains. This study will facilitate deeper understanding of the evolution of new variant CT and ToxT regulon, influencing cholera epidemiology.
High-dose rifamycins enable shorter oral treatment in a murine model of Mycobacterium ulcerans disease
Authors: Omansen Till F. , Almeida Deepak , Converse Paul J. , Li Si-Yang , Lee Jin , Stienstra Ymkje , van der Werf Tjip , Grosset Jacques H. , Nuermberger Eric L.
Published: August 2018

Buruli ulcer (BU), caused by Mycobacterium ulcerans, is a neglected tropical skin and soft tissue infection that is associated with disability and social stigma. The mainstay of BU treatment is an eight-week course of 10 mg/kg rifampin (RIF) and 150 mg/kg streptomycin (STR). Recently, the injectable STR has been shown to be replaceable with oral clarithromycin (CLR) for smaller lesions for the last four weeks of treatment. A shorter, all-oral, highly efficient regimen for BU is needed, as the long treatment duration and indirect costs currently burden patients and health systems. Increasing the dose of RIF or replacing it with the more potent rifamycin drug rifapentine (RPT) could provide such a regimen. Here, we performed a dose-ranging experiment of RIF and RPT in combination with CLR over four weeks of treatment in a mouse model of M. ulcerans disease. A clear dose-dependent effect of RIF on both clinical and microbiological outcomes was found, with no ceiling effect observed with tested doses up to 40 mg/kg. RPT-containing regimens were more effective on M. ulcerans. All RPT-containing regimens achieved culture negativity after only four weeks while only the regimen with the highest RIF dose (40 mg/kg) did so. We conclude that there is dose-dependent efficacy of both RIF and RPT and that a ceiling effect is not reached with the current standard regimen used in the clinic. A regimen based on higher rifamycin doses that are currently being evaluated against tuberculosis in clinical trials could shorten and improve therapy of Buruli ulcer.
Global identification of functional microRNA::mRNA interactions in Drosophila
Authors: Wessels Hans-Hermann , Lebedeva Svetlana , Hirsekorn Antje , Mukherjee Neelanjan , Ohler Uwe
Published: August 2018

MicroRNAs (miRNAs) are key mediators of post-transcriptional gene expression silencing. Although Drosophila has been of critical importance for miRNA discovery, biogenesis and function, there has been no comprehensive experimental annotation of functional miRNA target sites. To close this gap, we generated the first in vivo map of miRNA::mRNA interactions in Drosophila melanogaster, making use of crosslinked nucleotides in Argonaute (AGO) crosslinking and immunoprecipitation (CLIP) experiments that enable an unambiguous assignment of miRNAs to AGO binding sites at much higher signal-to-noise ratio than computational predictions alone.

Absolute quantification of cellular miRNA levels showed the miRNA pool in Drosophila cell lines to be more diverse than previously reported. Benchmarking two different CLIP approaches, we identified a similar predictive potential to unambiguously assign thousands of miRNA::mRNA pairs from AGO1 interaction data at unprecedented depth. Quantitative RNA-Seq and subcodon-resolution ribosomal footprinting data upon AGO1 depletion enabled the determination of miRNA-mediated effects on target expression and translation. We thus provide the first comprehensive resource of miRNA target sites as well as their quantitative functional impact in Drosophila.
Two-antibody pan-ebolavirus cocktail confers broad therapeutic protection in ferrets and nonhuman primates
Authors: Bornholdt Zachary A. , Herbert Andrew S. , Mire Chad E. , He Shihua , Cross Robert W. , Wec Anna , Abelson Dafna M. , Geisbert Joan B. , James Rebekah M. , Rahim Md Niaz , Zhu Wenjun , Borisevich Viktoriya , Banadyga Logan , Gunn Bronwyn M. , Agans Krystle N. , Goodwin Eileen , Tierney Kevin , Shestowsky William S. , Bohorov Ognian , Bohorova Natasha , Velasco Jesus , Ailor Eric , Kim Do , Pauly Michael H. , Whaley Kevin J. , Alter Galit , Walker Laura M. , Chandran Kartik , Zeitlin Larry , Qiu Xiangguo , Geisbert Thomas W. , Dye John M.
Published: August 2018

All available experimental vaccines and immunotherapeutics1,2 against Ebola virus (EBOV), including rVSV-ZEBOV3 and ZMappTM4, lack activity against other ebolaviruses associated with human disease outbreaks. This year, two separate outbreaks of EBOV in the Democratic Republic of Congo underscored the unpredictable nature of ebolavirus reemergence in a region that has historically experienced outbreaks of the divergent ebolaviruses Sudan virus (SUDV) and Bundibugyo virus (BDBV)5. Here we show that MBP134AF, a pan-ebolavirus therapeutic comprising two broadly neutralizing human antibodies (bNAbs)6,7(see companion manuscript, Wec et al.) could protect against lethal EBOV, SUDV, and BDBV infection in ferrets and nonhuman primates (NHPs). MBP134AF not only not only establishes a viable therapeutic countermeasure to outbreaks caused by antigenically diverse ebolaviruses but also affords unprecedented effectiveness and potency—a single 25-mg/kg dose was fully protective in NHPs. This best-in-class antibody cocktail is the culmination of an intensive collaboration spanning academia, industry and government in response to the 2013-2016 EBOV epidemic6,7 and provides a translational research model for the rapid development of immunotherapeutics targeting emerging infectious diseases.
Streptococcal superantigen-induced expansion of human tonsil T cells leads to altered T follicular helper cell phenotype, B cell death, and reduced immunoglobulin release
Authors: Davies Frances J. , Olme Carl , Lynskey Nicola N. , Turner Claire E. , Sriskandan Shiranee
Published: August 2018

Background

Streptococcal pyrogenic exotoxin (SPE)A expression is epidemiologically linked to streptococcal tonsillo-pharyngitis and outbreaks of scarlet fever, although the mechanisms by which superantigens confer advantageStreptococcus pyogenesare unclear.S. pyogenesis an exclusively human pathogen. As the leukocyte profile of tonsil differs from peripheral blood, the impact of SPEA production on human tonsil cell function was investigated.

Methods

Human tonsil cells from routine tonsillectomy, were co-incubated with purified streptococcal superantigens or isogenic streptococcal culture supernatants, differing only in superantigen content. Tonsil cell proliferation was quantified by tritium-incorporation, and cell surface characteristics assessed by flow-cytometry. Soluble mediators were measured using ELISA and quantitative (q)RT-PCR was performed for immunoglobulin gene expression.

Results

Tonsil T cells proliferated in response to SPEA and demonstrated typical release of pro-inflammatory cytokines. When cultured in the absence of superantigen, tonsil preparations released large quantities of immunoglobulin over 7d. In contrast, marked B cell apoptosis and abrogation of total IgA, IgM, and IgG production occurred in the presence of SPEA and other superantigens. In SPEA-stimulated cultures, T follicular helper (TfH) cells showed a reduction in CXCR5 expression, but up-regulation of CD134 (OX40), CD278 (ICOS) and CD150 (SLAM) expression, indicative of a phenotypic change in the TfH population and associated with impaired chemotactic response to CXCL13.

Conclusions

SPEA and other superantigens cause dysregulated tonsil immune function, driving T cells from TFH to a proliferating phenotype, with resultant loss of B cells and immunoglobulin production, providing superantigen-producing bacteria with a likely survival advantage.
Empirical Analysis and Modeling of Argos Doppler Location Errors in Romania
Authors: Rozylowicz Laurentiu , Bodescu Florian P. , Ciocanea Cristiana M. , Gavrilidis Athanasios A. , Manolache Steluta , Matache Marius L. , Miu Iulia V. , Moale Ionut C. , Nita Andreea , Popescu Viorel D.
Published: August 2018

Background

Advances in wildlife tracking technology have allowed researchers to understand the spatial ecology of many terrestrial and aquatic animal species. Argos Doppler is a technology that is widely used for wildlife tracking owing to the small size and low weight of the Argos transmitters. This allows them to be fitted to small-bodied species. The longer lifespan of the Argos units in comparison to units outfitted with miniaturized global positioning system (GPS) technology has also recommended their use. In practice, large Argos location errors often occur due to communication conditions such as transmitter settings, local environment, and the behavior of the tracked individual.

Methods

Considering the geographic specificity of errors and the lack of benchmark studies in Eastern Europe, the research objectives were: (1) to evaluate the accuracy of Argos Doppler technology under various environmental conditions in Romania, (2) to investigate the effectiveness of straightforward destructive filters for improving Argos Doppler data quality, and (3) to provide guidelines for processing Argos Doppler wildlife monitoring data. The errors associated with Argos locations in four geographic locations in Romania were assessed during static, low-speed and high-speed tests. The effectiveness of the Douglas Argos distance angle filter algorithm was then evaluated to ascertain its effect on the minimization of localization errors.

Results

Argos locations received in the tests had larger associated horizontal errors than those indicated by the operator of the Argos system, including under ideal reception conditions. Positional errors were similar to those obtained in other studies outside of Europe. The errors were anisotropic, with larger longitudinal errors for the vast majority of the data. Errors were mostly related to speed of the Argos transmitter at the time of reception, but other factors such as topographical conditions and orientation of antenna at the time of the transmission also contributed to receiving low-quality data. The Douglas-Argos filter successfully excluded the largest errors while retaining a large amount of data when the threshold was set to the local scale (2 km).

Discussion

Filter selection requires knowledge about the movement patterns and behavior of the species of interest, and the parametrization of the selected filter typically requires a trial and error approach. Selecting the proper filter reduces the errors while retaining a large amount of data. However, the post-processed data typically includes large positional errors; thus, we recommend incorporating Argos error metrics such as error ellipse or use complex modeling approaches when working with filtered data.
A human cell model of cardiac pathophysiological valvulogenesis
Authors: Neri Tui , Hiriart Emilye , Vliet Patrick van , Faure Emilie , Norris Russell A , Farhat Batoul , Lefrancois Julie , Moore-Morris Thomas , Zaffran Stéphane , Faustino Randolph S. , Zambon Alexander C , Sugi Yukiko , Desvignes Jean-Pierre , Salgado David , Levine Robert A. , de la Pompa Jose Luis , Terzic André , Evans Sylvia M. , Markwald Roger , Pucéat Michel
Published: August 2018

Genetically modified mice have advanced our understanding of valve development and related pathologies. Yet, little is known regarding human valvulogenesis in health and diseases. Genuine humanin vitromodels that reproduce valvular (patho)biology are thus needed. We here developed a human pluripotent stem cell-derived model fit to decode the early steps of human valvulogenesis and to recapitulate valve disease traits in a dish.

Using cellular based, single cell omics-informed andin vivo-validated approaches, we derived a population of pre-valvular endocardial cells from a pluripotent stem cell source. These human prevalvular cells (HPVCs) expressed gene patterns conforming to the atrio-ventricular canal (AVC) endocardium signature originally established in E9.0 mouse embryos. In fact, HPVC treated with BMP2, cultured onto mouse AVC cushions, or transplanted into the AVC of embryonic mouse hearts, underwent endothelial-to-mesenchymal transition and expressed markers of valve interstitial cells of different valvular layers demonstrating tissue functionality. HPVCs also differentiated into tendinous/chondrogenic cells in line with the valvular repertoire. Extending this valvulogenic model to patient specific iPS cells, we recapitulated features of mitral valve prolapse and uncovered that dysregulation of the SHH pathway is likely to be at the origin of the disease thus providing a putative therapeutic target.

Human pluripotent stem cells recapitulate early valvulogenesis and provide a powerful model to systematically decipher the origin and lineage contribution of different valvular cell types in humans as well as to study valve diseases in a dish.
Gene Correction for SCID-X1 in Long-Term Hematopoietic Stem Cells
Authors: Pavel-Dinu Mara , Wiebking Volker , Dejene Beruh T. , Srifa Waracharee , Mantri Sruthi , Nicolas Carmencita E. , Lee Ciaran , Bao Gang , Kildebeck Eric J. , Punjya Niraj , Sindhu Camille , Inlay Matthew A. , Saxena Nivi , DeRavin Suk See , Malech Harry , Roncarolo Maria Grazia , Weinberg Kenneth I. , Porteus Matthew H.
Published: August 2018

Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. High frequencies of reproducible targeted integration of a wild-type cDNA into the endogenous start codon of a gene in LT-HSCs could provide a robust genome editing approach to cure genetic diseases in which patients have different mutations throughout the gene. We describe a clinically relevant method for correcting X-linked severe combined immunodeficiency (SCID-X1). By using a highly specific and active CRISPR/Cas9-AAV6 based strategy and selection-free approach, we achieve up to 20% genome integration frequencies in LT-HSCs of a full-lengthIL2RGcDNA at the endogenous start site as demonstrated by serial transplantation and analysis of genome edited human cells eight months following initial transplantation. In addition to high frequencies of functional gene correction in LT-HSCs we observed no evidence of abnormal hematopoiesis following transplantation, a functional measure of the lack of genotoxicity. Deep analysis of potential off-target activity detected two sites with low frequency (<0.3%) of off-target mutations. The level of off-target mutations was reduced to below the limit of detection using a high fidelity Cas9. Moreover, karyotype evaluation identified no genomic instability events. We achieved high levels of genome targeting frequencies (median 45%) in CD34+HSPCs from six SCID-X1 patients and demonstrate rescue of lymphopoietic defect of patient derived cells bothin vitroandin vivo.In sum, our study provides specificity, toxicity and efficacy data supportive of clinical development of genome editing to treat SCID-Xl.
Rapid environmental change in games: complications and curiosities
Authors: Trimmer Pete C , Barrett Brendan J , McElreath Richard , Sih Andrew
Published: August 2018

Human-induced rapid environmental change (HIREC) has recently led to alterations in the fitness and behavior of many organisms. Game theory is an important tool of behavioral ecology for analyzing evolutionary situations involving multiple individuals. However, game theory bypasses the details by which behavioral phenotypes are determined, taking the functional perspective straight from expected payoffs to predicted frequencies of behaviors. In contrast with optimization approaches, we identify that to use existing game theoretic models to predict HIREC effects, additional mechanistic details (or assumptions) will often be required. We illustrate this in relation to the hawk-dove game by showing that three different mechanisms, each of which support the same ESS prior to HIREC (fixed polymorphism, probabilistic choice, or cue dependency), can have a substantial effect on behavior (and success) following HIREC. Surprisingly, an increase in the value of resources can lead to a reduction in payoffs (and vice versa), both in the immediate- and long-term following HIREC. An increase in expected costs also increases expected payoffs. Along with these counter-intuitive findings, this work shows that simply understanding the behavioral payoffs of existing games is insufficient to make predictions about the effects of HIREC.

It’s the little details that are vital. Little things make big things happen.

John Wooden
Stochastic population dynamics driven by mutant interactors
Authors: Park Hye Jin , Pichugin Yuriy , Huang Weini , Traulsen Arne
Published: August 2018

Spontaneous random mutations are an important source of variation in populations. Many evolutionary models consider mutants with a fixed fitness chosen from a certain fitness distribution without considering any interactions among the residents and mutants. Here, we go beyond this and consider “mutant interactors”, which lead to new interactions between the residents and invading mutants that can affect the carrying capacity and the extinction risk of populations. We model microscopic interactions between individuals by using a dynamical payoff matrix and analyze the stochastic dynamics of such populations. New interactions drawn from invading mutants can drive the population away from the previous equilibrium, and lead to changes in the population size — the population size is an evolving property rather than a fixed number or externally controlled variable. We present analytical results for the average population size over time and quantify the extinction risk of the population by the mean time to extinction.
TCF7L2 Regulation of GATA6-dependent and -Independent Vascular Smooth Muscle Cell Plasticity and Intimal Hyperplasia
Authors: Srivastava Roshni , Rolyan Harshavardhan , Xie Yi , Li Na , Bhat Neha , Hong Lingjuan , Esteghamat Fatemehsadat , Adeniran Adebowale , Geirsson Arnar , Zhang Jiasheng , Ge Guanghao , Nobrega Marcelo , Martin Kathleen A. , Mani Arya
Published: August 2018

Genetic variations in Wnt-coreceptor LRP6 and Wnt-regulated transcription factor TCF7L2 have been among the strongest genetic signals for type2 diabetes (T2DM) and coronary artery disease (CAD). Mice with a CAD-linked LRP6 mutation exhibit obstructive coronary artery disease characterized by reduced TCF7L2 expression and dedifferentiation of vascular smooth muscle cell (VSMC). While TCF7L2 maintains stemness and promotes proliferation in embryonic tissues and adult stem cells, its role and mechanisms of action in VSMC differentiation is not understood. Using multiple mouse models, we demonstrate here that TCF7L2 promotes differentiation and inhibits proliferation of VSMCs. TCF7L2 accomplishes these effects by stabilization of GATA6 and upregulation of SM-MHC and cell cycle inhibitors. Accordingly, TCF7L2 haploinsufficient mice exhibited increased susceptibility to, while mice overexpressing TCF7L2 were protected against injury-induced intimal hyperplasia compared to wildtype littermates. Consequently, the overexpression of TCF7L2 in LRP6 mutant mice rescued the injury induced intimal hyperplasia. These novel findings imply cell type-specific functional role of TCF7L2 and provide critical insight into poorly understood mechanisms underlying pathogenesis of intimal hyperplasia.
Neuraminidase Inhibition Contributes to Influenza A Virus Neutralization by Anti-Hemagglutinin Stem Antibodies
Authors: Kosik Ivan , Angeletti Davide , Gibbs James S. , Angel Matthew , Takeda Kazuyo , Kosikova Martina , Nair Vinod , Hickman Heather D. , Xie Hang , Brooke Christopher C. , Yewdell Jonathan W.
Published: August 2018

Broadly neutralizing antibodies (Abs) that bind the influenza virus hemagglutinin (HA) stem may enable universal influenza vaccination. Here, we show that anti-stem Abs sterically inhibit viral neuraminidase activity against large substrates, with activity inversely proportional to the length of the fibrous NA stalk that supports the enzymatic domain. By modulating NA stalk length in recombinant IAVs, we show that anti-stem Abs inhibit virus release from infected cells by blocking NA, accounting for theirin vitroneutralization activity. NA inhibition contributes to anti-stem Ab protection in influenza infected mice, likely due at least in part to NA-mediated inhibition of FcγR dependent activation of innate immune cells by antibody bound to virions. FDA approved NA inhibitors enhance anti-stem based Fcγdependent immune cell activation, raising the possibility of therapeutic synergy between NA inhibitors and anti-stem mAb treatment in humans.
Phase Transitioned Nuclear Oskar Promotes Cell Division of Drosophila Primordial Germ Cells
Authors: Kistler Kathryn E. , Trcek Tatjana , Hurd Thomas R. , Chen Ruoyu , Liang Feng-Xia , Sall Joseph , Kato Masato , Lehmann Ruth
Published: August 2018

Germ granules are non-membranous ribonucleoprotein granules deemed the hubs for post-transcriptional gene regulation and functionally linked to germ cell fate across species. Little is known about the physical properties of germ granules and how these relate to germ cell function. Here we study two types of germ granules in theDrosophilaembryo: cytoplasmic germ granules that instruct primordial germ cells (PGCs) formation and nuclear germ granules within early PGCs with unknown function. We show that cytoplasmic and nuclear germ granules are phase transitioned condensates nucleated by Oskar protein that display liquid as well as hydrogel-like properties. Focusing on nuclear granules, we find that Oskar drives their formation in heterologous cell systems. Multiple, independent Oskar protein domains synergize to promote granule phase separation. Deletion of Oskar’s nuclear localization sequence specifically ablates nuclear granules in cell systems. In the embryo, nuclear germ granules promote germ cell divisions thereby increasing PGC number for the next generation.