bluuebunny/crossref_metadata_2025 huggingface dataset:
Zinc finger of Arabidopsis thaliana 6 is involved in melatonin‐mediated auxin signaling through interacting INDETERMINATE DOMAIN15 and INDOLE‐3‐ACETIC ACID 17
Authors: Shi Haitao , Zhang Shengmin , Lin Daozhe , Wei Yunxie , Yan Yu , Liu Guoyin , Reiter Russel J. , Chan Zhulong
Published: April 2018

Although accumulating evidence demonstrates the cross talk between melatonin and auxin as derivatives of tryptophan, the underlying signaling events remain unclear. In this study, we found that melatonin and auxin mediated the transcriptional levels of zinc finger of Arabidopsis thaliana (ZAT6) in a mutually antagonistic manner. ZAT6 negatively modulated the endogenous auxin level, and ZAT6 knockdown plants were less sensitive to melatonin‐regulated auxin biosynthesis, indicating its involvement in melatonin‐mediated auxin accumulation. Additionally, the identification of INDETERMINATE DOMAIN15 (IDD15) and INDOLE‐3‐ACETIC ACID 17 (IAA17) in Arabidopsis that interacted with ZAT6 in vivo provided new insight of ZAT6‐mediated auxin signaling. Further investigation showed that ZAT6 repressed the transcription activation of IDD15 on the YUC2 promoter, while ZAT6 inhibited the interaction of TRANSPORT INHIBITOR RESPONSE 1 (TIR1) and IAA17 through competitively binding to IAA17. Thus, both auxin synthesis and the auxin response were negatively modulated by ZAT6. Taken together, ZAT6 is involved in melatonin‐mediated auxin signaling through forming an interacting complex of auxin signaling pathway in Arabidopsis.
Melatonin is involved in skotomorphogenesis by regulating brassinosteroid biosynthesis in rice plants
Authors: Hwang Ok Jin , Back Kyoungwhan
Published: April 2018

Serotonin N‐acetyltransferase (SNAT) is the penultimate enzyme in melatonin biosynthesis catalyzing the conversion of serotonin into N‐acetylserotonin. In plants, SNAT is encoded by 2 isogenes of which SNAT1 is constitutively expressed and its overexpression confers increased yield in rice. However, the role of SNAT2 remains to be clarified. In contrast to SNAT1, the diurnal rhythm of SNAT2 mRNA expression peaks at night. In this study, transgenic rice plants in which SNAT2 expression were suppressed by RNAi technology showed a decrease in melatonin and a dwarf phenotype with erect leaves, reminiscent of brassinosteroids (BR)‐deficient mutants. Of note, the dwarf phenotype was dependent on the presence of dark, suggesting that melatonin is involved in dark growth (skotomorphogenesis). In support of this suggestion, SNAT2 RNAi lines exhibited photomorphogenic phenotypes such as inhibition of internodes and increased expression of light‐inducible CAB genes in the dark. The causative gene for the melatonin‐mediated BR biosynthetic gene was DWARF4, a rate‐limiting BR biosynthetic gene. Exogenous melatonin treatment induced several BR biosynthetic genes, including DWARF4, D11, and RAVL1. As expected from the erect leaves, the SNAT2 RNAi lines produced less BR than the wild type. Our results show for the first time that melatonin is a positive regulator of dark growth or shade outgrowth by regulating BR biosynthesis in plants.
Multiphase emplacement of impact melt sheet into the footwall: Offset dykes of the Sudbury Igneous Complex, Canada
Authors: Prevec S. A. , Büttner S. H.
Published: April 2018

The offset dykes of the Sudbury Igneous Complex comprise two distinct main magmatic facies, a high‐temperature inclusion‐free quartz diorite (QD), and a subsequently intruded lower temperature, mineralized, and inclusion‐rich quartz diorite (MIQD). The MIQD facies was emplaced after QD dykes had solidified. Key controlling factors of the two injection phases were (1) the development of a coherent roof, which confined the melt sheet; and (2) the periodic increase of melt and fluid pressure within the melt sheet. For the injection of QD melt, the melt pressure exceeded the normal stress acting on fracture surfaces. For the later refracturing of QD dykes and the injection of MIQD melt, the melt pressure increased further, exceeding the tensile strength of, and the normal stress acting on, QD dykes. We associate the melt pressure increase required for both injection episodes with degassing and devolatilization of cooling melt close to the roof. Within the hydraulically connected melt column, the related pressure increase was transmitted to the base of the melt sheet where QD and MIQD melt was extracted into dykes. Residual core to rim thermal gradients in the QD dykes produced tensile strength gradients, accounting for the typically central location of MIQD dykes within QD dykes.
Formative research to explore the acceptability and use of infant food grinders for the promotion of animal source foods and micronutrient powders in rural Peru
Authors: Creed‐Kanashiro Hilary , Wasser Heather M. , Bartolini Rosario , Goya Cecilia , Bentley Margaret E.
Published: April 2018

According to global recommendations, quality diets for complementary feeding (CF) should include a diversity of foods including vitamin A‐rich fruits and vegetables and sources of high‐quality proteins and essential nutrients, particularly animal‐source foods (ASF). A key barrier to feeding ASF surrounds beliefs that the preparation of foods of a thicker consistency may cause problems of digestion, “heaviness” or stomach problems, swallowing, and choking. The objective of this study was to explore, through systematic formative research, the acceptability, use, and feasibility of a simple technology, commercial infant food grinders, in two rural Peruvian settings where there is delayed and low consumption of complementary foods of a thick consistency, including ASF. Phase I explored the barriers, constraints, and opportunities related to the provision of foods of a thicker consistency with a focus on ASF. Phase II encompassed household behavioural trials with mothers and infants to assess the acceptability and use of the grinders in the home setting, using key concepts and messages developed from the information obtained during Phase I. The technology was highly acceptable, used by the majority of mothers (87.8%), and led to changes in cultural perceptions, facilitating increased feeding of appropriate textures (thick purees), ASF, and multimicronutrient powders. Energy, protein, and micronutrient intakes were all significantly greater after the household behavioural trials. This simple technology, paired with systematic formative research to appropriately promote its use across cultures, may have a significant effect on improving CF practices globally, particularly for young infants beginning CF at 6 months.
Rebuilding the bridge between transcription and translation
Authors: Artsimovitch Irina
Published: April 2018

In Bacteria, ribosomes may bind to the nascent RNA emerging from the transcribing RNA polymerase and initiate translation. Transcription‐translation coupling plays diverse roles in cellular physiology, including attenuation control, mRNA surveillance and maintenance of genome integrity. While the existence of coupling is broadly accepted, its mechanism and ubiquity are debated. Structural evidence supports mutually exclusive modes of RNA polymerase‐ribosome contacts. In a model based on nuclear magnetic resonance data, NusG binds to a ribosomal protein S10 and acts as an adapter between RNA polymerase and the 30S subunit. Recent single‐particle cryo electron microscopy analyses of RNA polymerase bound to 30S and 70S ribosomes revealed extensive, and very distinct, contacts which are incompatible with bridging by NusG. Saxena et al. provide the first evidence for NusG‐mediated coupling in vivo. Their results demonstrate that Escherichia coli NusG interacts with the 70S ribosomes through a previously established interface and that these interactions are required for survival when translation elongation is hindered to weaken coupling. Future studies will address a likely possibility that distinct bridging mechanisms underpin context‐dependent coupling in the cell.
Clinical course of dengue fever and its impact on renal function in renal transplant recipients and patients with chronic kidney disease
Authors: Thomas E.T Arun , George Jacob , Sruthi Devi , Vineetha N. S. , Gracious Noble
Published: April 2018

Aim

Dengue fever is a mosquito‐borne viral disease endemic in many tropical and sub‐tropical countries. There are only limited data in the literature about dengue fever in renal transplant recipients and patients with chronic kidney disease. This study compares the clinical course of dengue fever and its impact on renal function in renal transplant recipients, patients with chronic kidney disease and patients with normal baseline renal function.

Methods

An observational study was conducted from 1 May to 31 July 2017, at a tertiary care centre of South India. A major epidemic of dengue had occurred during the study period. Twelve renal transplant recipients, 22 patients with CKD and 58 patients with normal baseline renal function (control group) admitted with dengue fever were prospectively studied.

Results

Nadir WBC count was lowest in renal transplant recipients (2575 ± 1187/mm3), [P < 0.001]. Renal transplant recipients took more time for normalization of platelet count (6 ± 4.5 days), [P < 0.001]. All 22 patients with CKD and 11 of 12 renal transplant recipients had worsening of renal function whereas only 17 of 58 patients in the control group had worsening [P < 0.001]. Sixteen patients with CKD, one renal transplant recipient and none among the control group required haemodialysis [P < 0.001]. Dialysis requiring patients had more haemoconcentration (52.5 ± 19.9% increase in haemoglobin), [P < 0.001]. Seven patients with CKD were dialysis dependent at the end of 2 weeks.

Conclusion

Clinical features of dengue fever were different in renal transplant recipients and patients with CKD. Severe worsening of renal function was common in CKD patients. Worsening of renal function in renal transplant recipients was less severe and transient.
Bedside nurses’ roles in discharge collaboration in general internal medicine: Disconnected, disempowered and devalued?
Authors: Goldman Joanne , MacMillan Kathleen , Kitto Simon , Wu Robert , Silver Ivan , Reeves Scott
Published: April 2018

Collaboration among nurses and other healthcare professionals is needed for effective hospital discharge planning. However, interprofessional interactions and practices related to discharge vary within and across hospitals. These interactions are influenced by the ways in which healthcare professionals’ roles are being shaped by hospital discharge priorities. This study explored the experience of bedside nurses’ interprofessional collaboration in relation to discharge in a general medicine unit. An ethnographic approach was employed to obtain an in‐depth insight into the perceptions and practices of nurses and other healthcare professionals regarding collaborative practices around discharge. Sixty‐five hours of observations was undertaken, and 23 interviews were conducted with nurses and other healthcare professionals. According to our results, bedside nurses had limited engagement in interprofessional collaboration and discharge planning. This was apparent by bedside nurses’ absence from morning rounds, one‐way flow of information from rounds to the bedside nurses following rounds, and limited opportunities for interaction with other healthcare professionals and decision‐making during the day. The disconnection, disempowerment and devaluing of bedside nurses in patient discharge planning has implications for quality of care and nursing work. Study findings are positioned within previous work on nurse–physician interactions and the current context of nursing care.
Nursing knowledge: A middle ground exploration
Authors: Sakamoto Mariko Liette
Published: April 2018

The discipline of nursing has long maintained that is has a unique contribution to make within the health care arena. This assertion of uniqueness lies in great part in the discipline's claim to a distinct body of knowledge. Nursing knowledge is characterized by diverse and multiple forms of knowing and underpins the work of all nurses, regardless of field of practice. Unfortunately, it has been challenging for the discipline to take full ownership of its epistemological diversity, largely due to factors such as competing worldviews, and ideological and binary positioning. A philosophical middle ground stance is proposed as a way for the discipline to contemplate, discuss and develop nursing knowledge; a middle space that provides the freedom to consider competing worldviews while still allowing for the discipline to fully express itself in all of its epistemological diversity. In being able to enact its multiple forms of knowledge in a creative and open space that is open to different ideas and worldviews, not only can nursing take full ownership of its practice and its unique knowledge, it can also demonstrate how best to navigate an increasingly polarized world. In a world that is increasingly fixated on binary solutions and dualistic points of view, it is time for nursing to celebrate its epistemological diversity.
Partisanship, Political Knowledge, and the Dunning‐Kruger Effect
Authors: Anson Ian G.
Published: April 2018

A widely cited finding in social psychology holds that individuals with low levels of competence will judge themselves to be higher achieving than they really are. In the present study, I examine how the so‐called “Dunning‐Kruger effect” conditions citizens' perceptions of political knowledgeability. While low performers on a political knowledge task are expected to engage in overconfident self‐placement and self‐assessment when reflecting on their performance, I also expect the increased salience of partisan identities to exacerbate this phenomenon due to the effects of directional motivated reasoning. Survey experimental results confirm the Dunning‐Kruger effect in the realm of political knowledge. They also show that individuals with moderately low political expertise rate themselves as increasingly politically knowledgeable when partisan identities are made salient. This below‐average group is also likely to rely on partisan source cues to evaluate the political knowledge of peers. In a concluding section, I comment on the meaning of these findings for contemporary debates about rational ignorance, motivated reasoning, and political polarization.
Ectopic expression of the Coffea canephora SERK1 homolog‐induced differential transcription of genes involved in auxin metabolism and in the developmental control of embryogenesis
Authors: Pérez‐Pascual Daniel , Jiménez‐Guillen Doribet , Villanueva‐Alonzo Hernán , Souza‐Perera Ramón , Godoy‐Hernández Gregorio , Zúñiga‐Aguilar José Juan
Published: April 2018

Somatic embryogenesis receptor‐like kinase 1 (SERK1) is a membrane receptor that might serve as common co‐regulator of plant cell differentiation processes by forming heterodimers with specific receptor‐like kinases. The Coffea canephora SERK1 homolog (CcSERK1) was cloned in this work, and its early function in the transcription of embryogenesis master genes and of genes encoding proteins involved in auxin metabolism was investigated by externally manipulating its expression in embryogenic leaf explants, before the appearance of embryogenic structures. Overexpression of CcSERK1 early during embryogenesis caused an increase in the number of somatic embryos when the 55‐day process was completed. Suppression of CcSERK1 expression by RNA interference almost abolished somatic embryogenesis. Real time‐PCR experiments revealed that the transcription of the CcAGL15, CcWUS, CcBBM, CcPKL, CcYUC1, CcPIN1 and CcPIN4 homologs was modified in direct proportion to the expression of CcSERK1 and that only CcLEC1 was inversely affected by the expression levels of CcSERK1. The expression of the CcYUC4 homolog was induced to more than 80‐fold under CcSERK1 overexpression conditions, but it was also induced when CcSERK1 expression was silenced. The level of CcTIR1 was not affected by CcSERK1 overexpression but was almost abolished during CcSERK1 silencing. These results suggest that CcSERK1 co‐regulates the induction of somatic embryogenesis in Coffea canephora by early activation of YUC‐dependent auxin biosynthesis, auxin transport mediated by PIN1 and PIN4, and probably auxin perception by the TIR1 receptor, leading to the induction of early‐stage homeotic genes (CcAGL15, CcWUS, CcPKL and CcBBM) and repression of late‐stage homeotic genes (CcLec1).
Brain‐heart interaction in perseverative cognition
Authors: Ottaviani Cristina
Published: April 2018

The move from the concept of homeostasis to that of allostasis has led reactivity stress research to widen the object of its investigation: from the brief physiological response that occurs when one is facing a stressor to what happens when one is anticipating or recovering from a stressor. A paradigmatic example is represented by perseverative cognition, during which human beings react “as if” they were constantly facing a concrete stressor. The core idea behind this review is that the cognitive inflexibility that characterizes perseverative cognition is reflected in both our body (by increased autonomic nervous system rigidity assessed by heart rate variability; HRV) and our brain (by reduced prefrontal‐amygdala functional connectivity). This is a review of studies conducted in different settings (laboratory, daily life), populations (healthy, major depression, generalized anxiety), location (United States, Europe), and age groups (children, adults) that consistently replicated the association between autonomic, subjective, and behavioral measures of cognitive inflexibility during perseverative cognition. Moreover, compelling neuroimaging data suggest that HRV reduction from pre‐ to post‐induction of perseverative cognition is associated with both structural and functional brain abnormalities reflecting impaired prefrontal inhibitory control over subcortical structures (e.g., diminished prefrontal‐amygdala functional connectivity). The integration of neuroscience techniques with clinical autonomic research has advanced our understanding of the neurobiology of brain–heart interaction during perseverative cognition, potentially yielding to more effective treatment packages. This is clinically relevant if one considers that perseverative cognition is a pervasive transdiagnostic factor that carries prognostic risk for both psychological and somatic health.
Fish community composition and diversity at restored estuarine habitats in Tampa Bay, Florida, United States
Authors: Schaberg Samantha J. , Patterson Joshua T. , Hill Jeffrey E. , Guindon Kathryn Y. , Tuckett Quenton M.
Published: April 2018

Estuary restoration in Tampa Bay, Florida, United States, is an ongoing focus of natural resource managers because of pressure from an increasing coastal population, historic habitat loss, and restoration's importance to economic development, recreational activities, and fish habitat. A growing population can also limit future large‐scale restorations due to associations with cost and land availability. This limitation might be overcome by applying the habitat mosaic approach to restoration, which creates distinct habitat types at small spatial scales. This approach was applied to create three types of estuarine habitat, reconnected tidal creek, salt marsh, and tidal pond. The objectives of this study were to (1) initiate monitoring of a restored wetland mosaic and (2) determine how fish diversity and community structure vary among restored habitat types. Replicated sampling using a 3‐mm mesh seine was used to characterize the fish communities. Our results indicate that the habitat mosaic approach creates suitable habitat for a variety of fish species where 37% of fish species were captured in just one habitat type. In particular, the recreationally important Centropomus undecimalis (common snook) was more common in the mangrove‐lined creek and the non‐native Sarotherodon melanotheron (blackchin tilapia) was common in the tidal pond. Greater emphasis should be placed on applied restoration research to identify how habitat types within a larger restoration mosaic contribute to local species diversity and recreationally and commercially important fishes, while limiting non‐natives. This emphasis could reveal how restoration approaches can be modified to include habitat mosaics, maximizing their contribution to productive fish habitat.
My possessions need me: Anthropomorphism and hoarding
Authors: Burgess Alexandra M. , Graves Lucy M. , Frost Randy O.
Published: April 2018

Hoarding disorder (HD), a new DSM‐5 classification, is characterized by difficulty discarding and the excessive acquisition of possessions to the extent that living spaces are compromised by clutter. Individuals with hoarding difficulties have a variety of motivations for object ownership, including emotional attachment towards their possessions which sometimes manifests through imbuing possessions with human‐like terms. Limited extant evidence suggests that anthropomorphism, attributing human qualities to non‐human objects, is related to hoarding, possibly because of difficulties with interpersonal attachment and social isolation. The current study investigated the relationship between hoarding behaviors (i.e., difficulty discarding, excessive acquisition, and clutter), hoarding beliefs (i.e., motivations for ownership including responsibility, emotional attachment, memory, control), anthropomorphism (i.e., generally in childhood, generally in adulthood, and towards three different personally‐owned objects), and loneliness. Moderation analyses examined whether hoarding beliefs or loneliness impacted how anthropomorphism related to hoarding symptoms. Results suggested that all dimensions of anthropomorphism were related to hoarding behaviors. Regression analyses indicated that anthropomorphism in adulthood and of personally owned‐objects were the best predictors of hoarding behavior. Mixed evidence was found for hoarding beliefs and loneliness moderating these associations. Findings successfully replicated and extended previous literature and provide a novel measure of anthropomorphism specifically incorporating personal ownership.
Protest Movements and Citizen Discontent: Occupy Wall Street and the Tea Party
Authors: Martin John Levi , Murphy James P. , Moore Rick
Published: April 2018

Why do citizens indicate support for protest movements such as the Tea Party and Occupy Wall Street? There have been two general sets of explanations. One set emphasizes that support comes from those for whom the existing party system, and the ideological differentiation that corresponds to party divisions, are irrelevant. The second set takes the opposite tack, and emphasizes that the only thing that supporters of protests movements find lacking in the party system is extremity. Using some underexplored data, we present evidence that both accounts are incorrect for the case of these recent movements (Occupy and the Tea Party): what provokes support for protest movements is not ideology itself but a fundamental rejection of the current state of the party system, which we call disgruntlement. What ideology does for supporters is provide a sense of political friends and enemies (or near and far), which then can channel the direction that this disgruntlement takes. Further, ideologues with more education are more resistant to the appeal of the protest movement associated with the other political camp.
Becoming us and them: Social learning and intergroup bias
Authors: Over Harriet , McCall Cade
Published: April 2018

In recent years, research has demonstrated that the basic features of prejudice and discrimination emerge early in children's development. These discoveries call into question the role of social learning in intergroup bias. Specifically, through what means do we learn to distinguish “us” from “them?” Here, we explore this question, focusing on three key issues: how children respond to biased information they receive from others, how children selectively seek out certain types of biased information, and how children communicate biased information to others. We close by discussing the implications of this research for interventions to reduce stereotyping, prejudice, and discrimination.
Towards holomorphology in entomology: rapid and cost‐effective adult–larva matching using NGS barcodes
Authors: Yeo Darren , Puniamoorthy Jayanthi , Ngiam Robin Wen Jiang , Meier Rudolf
Published: April 2018

In many taxa the morphology of females and immatures is poorly known because species descriptions and identification tools have a male bias. The root causes are problems with matching life‐history stages and genders belonging to the same species. Such matching is time‐consuming when conventional methods are used (e.g. rearing) and expensive when the stages are matched with DNA barcodes. Unfortunately, the lack of associations is not a trivial problem because it renders a large part of the phenome of insects unexplored, although larvae and females are useful sources of characters for descriptive and phylogenetic purposes. In addition, many collectors intentionally avoid females and immature stages, which skews survey results, interferes with collecting life‐history information, and makes it less likely that rare species are discovered. These problems even exist for well‐studied taxa like Odonata, where obtaining adult–larva matches relies largely on rearing. Here we demonstrate how the matching problem can be addressed with cost‐effective tagged amplicon sequencing of a 313‐bp segment of cox1 with next‐generation sequencing (NGS) (‘NGS barcoding’). We illustrate the value of this approach based on Singapore's odonate fauna which is of a similar size as the European fauna (Singapore, 122 extant species; Europe, 138 recorded species). We match the larvae and adults of 59 species by first creating a barcode database for 338 identified adult specimens representing 83 species. We then sequence 1178 larvae from a wide range of sources. We successfully barcode 1123 specimens, which leads to adult–larva matches for 59 species based on our own barcodes (55) and online barcode databases (4). With these additions, 84 of the 131 species recorded in Singapore have now been associated with a species name. Most common species are now matched (83%), and good progress has been made for vulnerable/near‐threatened (55%), endangered (53%), and critically endangered species (38%). We used nondestructive DNA extraction methods in order to be able to use high‐resolution imaging of matched larvae to establish a publicly available digital reference collection for odonates which is incorporated into ‘Biodiversity of Singapore’ (
https://singapore.biodiversity.online/). We suggest that the methods described here are suitable for many insect taxa because NGS barcoding allows for fast and low‐cost matching of well‐studied life‐history stages with neglected semaphoronts (eggs, larvae, females). We estimate that the specimen‐specific amplicons in this study (c. 1500 specimens) can now be obtained within eight working days and that the laboratory and sequencing cost is c. US$600 (< US$0.40 per specimen).
Massive transfusions for critical bleeding: is everything old new again?
Authors: Flint A.W.J. , McQuilten Z. K. , Wood E. M.
Published: April 2018

Massive transfusion or major haemorrhage protocols have been widely adopted in the treatment of critically bleeding patients. Following evidence that higher ratios of transfused plasma and platelets to red blood cells may offer survival benefits in military trauma patients, these ratios are now commonly incorporated into massive transfusion protocols. They more closely resemble the effects of whole blood transfusion, which in the second half of last century was largely replaced by individual blood component transfusion based on laboratory‐guided indicators. However, high‐quality evidence to guide transfusion support for critically bleeding patients across the range of bleeding contexts is lacking, including for both trauma and non‐trauma patients. More data on major haemorrhage support and clinical outcomes are needed to inform guidelines and practice.
Impact of bacterial translocation in calves with atresia coli
Authors: Altan Semih , Koç Yılmaz , Alkan Fahrettin , Sayın Zafer , Erol Muharrem
Published: April 2018

Objective

To identify whether enteric bacteria pass into the mesenteric lymph nodes (MLNs) and peritoneal cavity in calves with atresia coli and to evaluate whether the presence of bacterial translocation (BT) has an impact on the success of surgical treatment.

Design

Prospective clinical study.

Animals

Twenty‐six client‐owned calves.

Interventions

During laparotomy, swab samples were collected from the peritoneal cavity and MLNs using a sterile swab stick and were submitted for microbiological analysis.

Measurements and Main Results

Bacterial cultures of swab samples revealed that 65% (n= 17) of the calves experienced BT. Of these, 14 calves experienced BT to the MLNs, 9 to the peritoneal cavity, and 5 to both regions. Of the bacteria isolated from the MLNs, 72% (n = 10) wereEscherichia coli. Of the samples isolated from the peritoneal fluid, 33% (n = 3) containedE. coliand 33% (n = 3) containedE. coli+ coagulase‐negativeStaphylococcus(CNS). In calves with BT that were discharged (n = 13) and without BT that were discharged (n = 7), the median survival was 30 days; these data were found to be similar in the 2 groups.

Conclusions

This study revealed that BT is observed in the majority of atresia coli cases.E. coliis more common in BT, and translocation occurs primarily through the lymphatic route. These results suggest that the presence of BT is closely related to the success of the operation for correction of atresia coli.
Regulation of muscle protein synthesis in an in vitro cell model using ex vivo human serum
Authors: Carson Brian P. , Patel Bijal , Amigo‐Benavent Miryam , Pauk Martina , Kumar Gujulla Sunil , Murphy Sylvia M. , Kiely Patrick A. , Jakeman Philip M.
Published: April 2018

New Findings

What is the central question of this study?

Can medium conditioned by ex vivo human serum regulate muscle protein synthesis in skeletal muscle cells in vitro?

What is the main finding and its importance?

This study demonstrates that medium conditioned by ex vivo human serum can regulate muscle protein synthesis in skeletal muscle cells in vitro via the mammalian Target of Rapomycin (mTOR) pathway, and this can be regulated differentially by fed and fasted ex vivo human serum.

Human serum embodies the integrated systemic response to any condition or perturbation, which may regulate muscle protein synthesis (MPS). Conditioning of medium with human serum represents a physiologically relevant method of regulating MPS in vitro. The primary purpose of the study was the development of a model using ex vivo human serum to condition medium and regulate MPS in in vitro skeletal muscle cells. Four young healthy men reported to the laboratory after an overnight fast and were fed with 0.33 g (kg body mass)−1 whey protein. Blood samples were taken before (Fasted) and 60 min postprandial (Fed). Fully differentiated C2C12 skeletal muscle cells were nutrient and serum deprived for 1 h and subsequently treated with medium conditioned with Fasted or Fed ex vivo human serum (20%) for 4 h. The MPS was measured using the surface sensing of translation technique and activation of mTOR, P70S6K and 4EBP1 by Western blot. Fasted and fed ex vivo human serum increased MPS (P < 0.05). Although a strong effect (ƞ2 = 0.36) for increased MPS in Fed relative to Fasted was observed, this was not statistically significant (P > 0.05). Activation of mTOR, P70S6K and 4EBP1 was significantly increased after treatment with Fed compared with Fasted ex vivo human serum (P < 0.05). Here, we developed and optimized the conditions for culture of C2C12 skeletal muscle cells, measurement of MPS and signalling in medium conditioned by ex vivo human serum. Furthermore, the functionality of the model was demonstrated by comparison of the response to medium conditioned by Fasted and Fed ex vivo human serum.
Evaluation of the Antifungal Activity of the Novel Oral Glucan Synthase Inhibitor SCY-078, Singly and in Combination, for the Treatment of Invasive Aspergillosis
Authors: Ghannoum M. , Long L. , Larkin E. L. , Isham N. , Sherif R. , Borroto-Esoda K. , Barat S. , Angulo D.
Published: April 2018

Invasive aspergillosis remains a major cause of death among the immunocompromised population and those receiving long-term immunosuppressive therapy. In light of increased azole resistance, variable outcomes with existing echinocandin monotherapy and combination therapy, and persistent high mortality rates, new antifungal agents for the treatment of invasive aspergillosis are clearly needed. SCY-078 is the first-in-class triterpenoid antifungal, a novel class of glucan synthase inhibitors with broad
in vitro
and
in vivo
activity against a broad spectrum of
Candida
and
Aspergillus
species.
In vitro
testing of clinical strains of
Aspergillus fumigatus
and non-
fumigatus Aspergillus
strains showed that SCY-078 had potent fungistatic activity (minimum effective concentration for 90% of strains tested = 0.125 μg/ml) compared with the activities of amphotericin B (MIC
90
= 8 μg/ml) and voriconazole (MIC
90
= 2 μg/ml). Testing of SCY-078 in combination with isavuconazole or voriconazole demonstrated synergistic activity against the majority of the azole-susceptible strains tested, and SCY-078 in combination with amphotericin B was synergistic against the azole-susceptible strains, as well as one known resistant
cyp51A
mutant. SCY-078 may be an important additional antifungal for first-line or salvage monotherapy or combination treatment of invasive aspergillosis.
Pyocyanin Inhibits Chlamydia Infection by Disabling Infectivity of the Elementary Body and Disrupting Intracellular Growth
Authors: Li Jian Lin , Yang Ningjing , Huang Lei , Chen Dandan , Zhao Yu , Tang M. Matt , Fan Huizhou , Bao Xiaofeng
Published: April 2018

The obligate intracellular bacteriumChlamydiais a widespread human pathogen that causes serious problems, including (but not limited to) infertility and blindness. Our search for novel antichlamydial metabolites from marine-derived microorganisms led to the isolation of pyocyanin, a small compound fromPseudomonas aeruginosa. Pyocyanin is an effective antichlamydial for all threeChlamydiaspp. tested. It has a 50% inhibitory concentration (IC50) of 0.019 to 0.028 μM, which is comparable to the IC50of tetracycline. At concentrations as low as 0.0039 μM, pyocyanin disables infectivity of the chlamydial elementary body (EB). At 0.5 μM or higher concentrations, the continuous presence of pyocyanin also inhibits chlamydial growth in the inclusion during later stages of the developmental cycle. Oxidative stress, a major known antimicrobial mechanism of pyocyanin, appears to be responsible only for the inhibition of bacterial growth and not for the disinfection of EBs. Pyocyanin is well-tolerated by probiotic vaginalLactobacillusspp. Our findings suggest that pyocyanin is of therapeutic value for chlamydial infections and can serve as a valuable chemical probe for studying chlamydial biology.
The Pyrroloquinoline-Quinone-Dependent Pyranose Dehydrogenase from Coprinopsis cinerea Drives Lytic Polysaccharide Monooxygenase Action
Authors: Várnai Anikó , Umezawa Kiwamu , Yoshida Makoto , Eijsink Vincent G. H.
Published: April 2018

Fungi secrete a set of glycoside hydrolases and oxidoreductases, including lytic polysaccharide monooxygenases (LPMOs), for the degradation of plant polysaccharides. LPMOs catalyze the oxidative cleavage of glycosidic bonds after activation by an external electron donor. So far, only flavin-dependent oxidoreductases (from the auxiliary activity [AA] family AA3) have been shown to activate LPMOs. Here, we present LPMO activation by a pyrroloquinoline-quinone (PQQ)-dependent pyranose dehydrogenase (PDH) fromCoprinopsis cinerea,CcPDH, the founding member of the recently discovered auxiliary activity family AA12.CcPDH contains a C-terminal family 1 carbohydrate binding module (CBM1), an N-terminal family AA8 cytochrome domain, and a central AA12 dehydrogenase domain. We have studied the ability of full-lengthCcPDH and its truncated variants to drive catalysis by twoNeurospora crassaLPMOs. The results show thatCcPDH indeed can activate the C-1-oxidizingN. crassaLPMO 9F (NcLPMO9F) and the C-4-oxidizingNeurospora crassaLPMO 9C (NcLPMO9C), that this activation depends on the cytochrome domain, and that the dehydrogenase and the LPMO reactions are strongly coupled. The two testedCcPDH-LPMO systems showed quite different efficiencies, and this difference disappeared upon the addition of free PQQ acting as a diphenol/quinone redox mediator, showing that LPMOs differ when it comes to their direct interactions with the cytochrome domain. Surprisingly, removal of the CBM domain fromCcPDH had a considerable negative impact on the efficiency of theCcPDH-LPMO systems, suggesting that electron transfer in the vicinity of the substrate is beneficial.CcPDH does not oxidize cello-oligosaccharides, which makes this enzyme a useful tool for studying cellulose-oxidizing LPMOs.

IMPORTANCELytic polysaccharide monooxygenases (LPMOs) are currently receiving increasing attention because of their importance in degrading recalcitrant polysaccharides and their potential roles in biological processes, such as bacterial virulence. LPMO action requires an external electron donor, and fungi growing on biomass secrete various so-called glucose-methanol-choline (GMC) oxidoreductases, including cellobiose dehydrogenase, which can donate electrons to LPMOs. This paper describes how an enzyme not belonging to the GMC oxidoreductase family,CcPDH, can activate LPMOs, and it provides new insights into the activation process by (i) describing the roles of individualCcPDH domains (a dehydrogenase, a cytochrome, and a carbohydrate-binding domain), (ii) showing that the PDH and LPMO enzyme reactions are strongly coupled, (iii) demonstrating that LPMOs differ in terms of their efficiencies of activation by the same activator, and (iv) providing indications that electron transferring close to the substrate surface is beneficial for the overall efficiency of theCcPDH-LPMO system.
Comparison of Uncleaved and Mature Human Immunodeficiency Virus Membrane Envelope Glycoprotein Trimers
Authors: Castillo-Menendez Luis R. , Witt Kristen , Espy Nicole , Princiotto Amy , Madani Navid , Pacheco Beatriz , Finzi Andrés , Sodroski Joseph
Published: April 2018

The mature envelope glycoprotein (Env) spike on the surfaces of human immunodeficiency virus type 1 (HIV-1)-infected cells and virions is derived from proteolytic cleavage of a trimeric gp160 glycoprotein precursor. In these studies, we compared the conformations of cleaved and uncleaved membrane Envs with truncated cytoplasmic tails to those of stabilized soluble gp140 SOSIP.664 Env trimers. Deletion of the gp41 cytoplasmic tail did not significantly affect the sensitivity of viruses with the HIV-1
AD8
Env to inhibition by antibodies or a CD4-mimetic compound. After glutaraldehyde fixation and purification from membranes, a cleaved Env exhibited a hydrodynamic radius of ∼10 nm and an antibody-binding profile largely consistent with that expected based on virus neutralization sensitivity. The purified cleaved Env trimers exhibited a hollow architecture with a central void near the trimer axis. Uncleaved Env, cross-linked and purified in parallel, exhibited a hydrodynamic radius similar to that of the cleaved Env. However, the uncleaved Env was recognized by poorly neutralizing antibodies and appeared by negative-stain electron microscopy to sample multiple conformations. Compared with membrane Envs, stabilized soluble gp140 SOSIP.664 Env trimers appear to be more compact, as reflected in their smaller hydrodynamic radii and negative-stain electron microscopy structures. The antigenic features of the soluble gp140 SOSIP.664 Env trimers differed from those of the cleaved membrane Env, particularly in gp120 V3 and some CD4-binding-site epitopes. Thus, proteolytic maturation allows the membrane-anchored Env to achieve a conformation that retains functional metastability but masks epitopes for poorly neutralizing antibodies.

IMPORTANCE
The entry of human immunodeficiency virus type 1 (HIV-1) into host cells is mediated by the envelope glycoprotein (Env) spike on the surface of the virus. Host antibodies elicited during natural HIV-1 infection or by vaccination can potentially recognize the Env spike and block HIV-1 infection. However, the changing shape of the HIV-1 Env spike protects the virus from antibody binding. Understanding the shapes of natural and man-made preparations of HIV-1 Envs will assist the development of effective vaccines against the virus. Here, we evaluate the effects of several Env modifications commonly used to produce Env preparations for vaccine studies and the determination of structure. We found that the cleavage of the HIV-1 Env precursor helps Env to assume its natural shape, which resists the binding of many commonly elicited antibodies. Stabilized soluble Envs exhibit more compact shapes but expose some Env elements differently than the natural Env.
Porcine Deltacoronavirus Engages the Transmissible Gastroenteritis Virus Functional Receptor Porcine Aminopeptidase N for Infectious Cellular Entry
Authors: Wang Bin , Liu Yan , Ji Chun-Miao , Yang Yong-Le , Liang Qi-Zhang , Zhao Pengwei , Xu Ling-Dong , Lei Xi-Mei , Luo Wen-Ting , Qin Pan , Zhou Jiyong , Huang Yao-Wei
Published: April 2018

Identification of cellular receptors used by coronavirus (CoV) entry into the host cells is critical to an understanding of pathogenesis and to development of intervention strategies. The fourth CoV genus,
Deltacoronavirus
, evolutionarily related to the
Gammacoronavirus
, has just been defined recently. In the current study, we demonstrate that porcine aminopeptidase N (pAPN) acts as a cross-genus CoV functional receptor for both enteropathogenic porcine deltacoronovirus (PDCoV) and alphacoronovirus (AlphaCoV) (transmissible gastroenteritis virus [TGEV]) based upon three lines of evidence. First, the soluble S1 protein of PDCoV bound to the surface of target porcine cell lines known to express pAPN as efficiently as TGEV-S1, which could be blocked by soluble pAPN pretreatment. Second, both PDCoV-S1 and TGEV-S1 physically recognized and interacted with pAPN by coimmunoprecipitation in pAPN cDNA-transfected cells and by dot blot hybridization assay. Finally, exogenous expression of pAPN in refractory cells conferred susceptibility to PDCoV-S1 binding and to PDCoV entry and productive infection. PDCoV-S1 appeared to have a lower pAPN-binding affinity and likely consequent lower infection efficiency in pAPN-expressing refractory cells than TGEV-S1, suggesting that there may be differences between these two viruses in the virus-binding regions in pAPN. This study paves the way for dissecting the molecular mechanisms of PDCoV-host interactions and pathogenesis as well as facilitates future vaccine development and intervention strategies against PDCoV infection.

IMPORTANCE
The emergence of new human and animal coronaviruses is believed to have occurred through interspecies transmission that is mainly mediated by a species-specific receptor of the host. Among the four genera of the
Coronavirinae
, a couple of functional receptors for the representative members in the genera
Alphacoronavirus
and
Betacoronavirus
have been identified, whereas receptors for
Gammacoronavirus
and
Deltacoronavirus
, which are believed to originate from birds, are still unknown. Porcine coronaviruses, including the newly discovered porcine deltacoronavirus (PDCoV) associated with diarrhea in newborn piglets, have posed a serious threat to the pork industry in Asia and North America. Here, we report that PDCoV employs the alphacoronavirus TGEV functional receptor porcine aminopeptidase N (pAPN) for cellular entry, demonstrating the usage of pAPN as a cross-genus CoV functional receptor. The identification of the PDCoV receptor provides another example of the expanded host range of CoV and paves the way for further investigation of PDCoV-host interaction and pathogenesis.
EF-P Posttranslational Modification Has Variable Impact on Polyproline Translation in
Bacillus subtilis
Authors: Witzky Anne , Hummels Katherine R. , Tollerson Rodney , Rajkovic Andrei , Jones Lisa A. , Kearns Daniel B. , Ibba Michael
Published: April 2018

Elongation factor P (EF-P) is a ubiquitous translation factor that facilitates translation of polyproline motifs. In order to perform this function, EF-P generally requires posttranslational modification (PTM) on a conserved residue. Although the position of the modification is highly conserved, the structure can vary widely between organisms. In
Bacillus subtilis
, EF-P is modified at Lys32 with a 5-aminopentanol moiety. Here, we use a forward genetic screen to identify genes involved in 5-aminopentanolylation. Tandem mass spectrometry analysis of the PTM mutant strains indicated that
ynbB
,
gsaB
, and
ymfI
are required for modification and that
yaaO
,
yfkA
, and
ywlG
influence the level of modification. Structural analyses also showed that EF-P can retain unique intermediate modifications, suggesting that 5-aminopentanol is likely directly assembled on EF-P through a novel modification pathway. Phenotypic characterization of these PTM mutants showed that each mutant does not strictly phenocopy the
efp
mutant, as has previously been observed in other organisms. Rather, each mutant displays phenotypic characteristics consistent with those of either the
efp
mutant or wild-type
B. subtilis
depending on the growth condition.
In vivo
polyproline reporter data indicate that the observed phenotypic differences result from variation in both the severity of polyproline translation defects and altered EF-P context dependence in each mutant. Together, these findings establish a new EF-P PTM pathway and also highlight a unique relationship between EF-P modification and polyproline context dependence.

IMPORTANCE
Despite the high level of conservation of EF-P, the posttranslational modification pathway that activates EF-P is highly divergent between species. Here, we have identified and characterized in
B. subtilis
a novel posttranslational modification pathway. This pathway not only broadens the scope of potential EF-P modification strategies, but it also indicates that EF-P modifications can be assembled directly on EF-P. Furthermore, characterization of these PTM mutants has established that an altered modification state can impact both the severity of polyproline translational defects and context dependence.