bluuebunny/crossref_metadata_2025 huggingface dataset:
Food repertoire history in children with autism spectrum disorder in Japan
Authors: Tanoue Koji , Takamasu Tetsuya , Matsui Kiyoshi
Published: September 2016

Background

Food selectivity is commonly reported in children with autism spectrum disorder (ASD). The aim of this study was to investigate eating habit history in children with ASD.

Methods

We analyzed 3 day food records completed by the parents and assessed how many unique foods each child consumed. The parents were also interviewed about their child's diet of complementary (i.e. transition) foods and estimated food repertoire at the ages of 3, 6, 12 and 18 years.

Results

A total of 28 participants were enrolled in this study. Some participants had ongoing changes in food repertoire from the age of 3 years onward. In two cases, although the number of foods consumed at age 3 years was approximately 50, this decreased markedly, becoming severely limited, by age 5 years. One of the reasons for diminished repertoire was infection, such as acute gastroenteritis and upper respiratory tract infection. In contrast, five patients had a severely limited food repertoire at age 3 years, which later increased to 15 or more. Four patients had good opportunity at school to increase their food repertoire.

Conclusions

Diet history varied and changed in response to new opportunities, education and/or the environment. In some cases the number of foods consumed decreased gradually due to anxiety and stress, resulting in a severely limited food repertoire. Some patients had good opportunities to increase their repertoire at school. If an effective program in the early years achieves progress, the eating habits of children with ASD might be changed.
Do Not Revise Ockham's Razor Without Necessity
Authors: Baron Sam , Tallant Jonathan
Published: September 2016

Ockham's razor asks that we not multiply entities beyond necessity. The razor is a powerful methodological tool, enabling us to articulate reasons for preferring one theory to another. There are those, however, who would modify the razor. Schaffer (2010: 313—our italics), for one, tells us that, ‘I think the proper rendering of Ockham's razor should be ‘Do not multiplyfundamentalentities without necessity’’. Our aim, here, is to challenge such re‐workings of Ockham's razor.
Open Data for Science, Policy, and the Public Good
Authors: Sá Creso , Grieco Julieta
Published: September 2016

Supporters of open data believe that free and complete access to research data is beneficial for science, public policy, and society. In environmental science and policy, open data systems can enable relevant research and inform evidence‐based governmental decisions. This article examines the unlikely case of Brazil's National Institute for Space Research's transition toward an open data model. Considering Brazil's young democracy, incipient practice of government transparency and accountability, and lacking a tradition of science‐policy dialogue, this case is a striking example of how open data can support public debate by making information about forest cover widely available. The case shows the benefits and challenges of developing such open data systems, and highlights the various forms of accessibility involved in making data available to the public.
Measuring the Real and Financial Connectedness of Selected African Economies with the Global Economy
Authors: Ogbuabor Jonathan E. , Orji Anthony , Aneke Gladys C. , Erdene‐Urnukh Oyun
Published: September 2016

We examine the real and financial connectedness of selected African economies with the global economy using a network approach. We find that the connectedness of African economies with the global economy is quite sizable, with the global financial crisis increasing the connectedness measures above their pre‐crisis levels. The results show that U.S., EU and Canada dominate Africa's equity markets, while China, India and Japan dominate Africa's real activities. Our results suggest that African economies are predominantly small open economies, deeply interconnected but systemically unimportant and vulnerable to headwinds emanating from the dominant economies in the overall global economy.
Fine‐grained meandering systems of the Lower Permian Clear Fork Formation of north‐central Texas, USA: Lateral and oblique accretion on an arid plain
Authors: Simon Sharane S.T. , Gibling Martin R.
Published: September 2016

Facies models that adequately represent the diverse range of fine‐grained fluvial systems are currently lacking from the literature. In this paper, the spectrum of these systems on the arid plains of western equatorial Pangea is explored, as well as the source and nature of the fine‐grained sediments. Eight fluvial elements in the Early Permian Clear Fork Formation of north‐central Texas represent channel systems up to 7 m deep with coarse basal deposits, three types of lateral‐accretion deposits and sandstone sheets, with laminated, disrupted and massive mudstones laid down in abandoned channels and on floodplains. The three fine‐grained fluvial styles represent a continuum between two end‐members: sustained lateral accretion of bedload composed of quartzose sediments and mud aggregates on point bars, and oblique accretion of suspended sediment on steep accretionary benches and banks with limited lateral migration. This spectrum is controlled, in part, by grain size and the proportion of suspended to bedload sediments. The presence of rarely documented swept ripples on exhumed accretion surfaces is attributed to rapid decline in water levels and downstream re‐entry of overbank floodwaters into the channel. Rill casts, roots and disrupted mudstones low down in channel bodies indicate periods of near‐dryness. Laterally extensive sheet sandstones were formed by episodic flows in broad, sandbed channels. The fluvial sediments were primarily intrabasinally sourced with extrabasinal sediments brought in during major floods from upland source areas or reworked from local storage in the basin, representing a supply limited system. The upward change in cement composition from mainly calcite and ankerite to dolomite and gypsum with minor celestine implies increasingly saline groundwater and progressive aridification, supporting Late Palaeozoic palaeoclimatic models. By integrating petrographic data with sedimentology, a plethora of information about ancient landscapes and climate is provided, allowing a fuller comparison between the Clear Fork Formation and modern dryland alluvial plains.
A Contextualist Approach to Higher‐Order Vagueness
Authors: Montminy Martin
Published: September 2016

According to contextualism about vagueness, the content of a vague predicate is context sensitive. On this view, when item a is in the penumbra of the vague predicate ‘F’, speakers may (truly) utter ‘Fa’, or they may (truly) utter ‘not‐Fa’, without contravening the literal meaning of ‘F’. Unlike its more popular variants, the version of contextualism I defend rejects the principle of tolerance, a principle according to which small differences should not affect the applicability of a vague predicate. My goal is to show how such a rejection allows for a plausible treatment of higher‐order vagueness, and for a dissolution of paradoxes of higher‐order vagueness.
Exploitation, Domination, Competitive Markets, and Unfair Division
Authors: Arneson Richard
Published: September 2016

When the assertion that some agent is exploiting a person connotes that the exploitation is morally wrong, what is this wrong? Some maintain that exploitation need not involve unfair division of advantages, but instead is essentially domination for self‐enrichment. This essay denies this claim and upholds the idea that exploitation claims concern unfair distribution. Some maintain that the hypothetical fully competitive market exchange price can serve, at least in some contexts, as the standard for assessing whether voluntary interaction is exploitative. This essay denies that the idea of the fully competitive market price can serve in this role. Nor should we accept a pure luck egalitarian claim that would identify fair distribution with the outcome of ideally competitive markets proceeding from fair initial distribution.
The Tension between the Nature and the Norm of Voluntary Exchange
Authors: Christiano Thomas
Published: September 2016

I develop a conception of voluntary exchange and its value that helps us understand the fundamental source of difficulty with voluntary exchange. We can make a great deal of progress in understanding the promise and the perils of voluntary exchange by elaborating an analogy between voluntary exchange and democracy. To be sure, this is a hazardous activity since there are many differences between these areas. But a careful effort here will illuminate the domain of voluntary exchange in both normative and descriptive dimensions. I argue that there is a fundamental tension between the normative principle that applies to voluntary exchange and the basic mechanism by which voluntary exchange operates. The fundamental normative principle, I will argue, is the principle that power over the making of an agreement with another ought to be proportioned among the persons to the interests each person has at stake in the making of the agreement. The fundamental mechanism of voluntary exchange, on the other hand, is that power is inversely proportioned to the stakes someone has in making the agreement. The more interests I have at stake in making an agreement, the more say over the content of the agreement I should have, and, yet, the more my interests hang on an agreement, the less bargaining power I have and so the less say I have in the making of the agreement. Hence, the inherent tendency of voluntary exchange is to work against the realization of the fundamental normative principle that regulates voluntary exchange. Hence, we have a deep and pervasive opposition between fact and norm in the very nature of voluntary exchange. All is not entirely hopeless, however, since there is a unique point where this opposition does not appear and that is in equality in the background conditions of exchange.
Using HHsearch to tackle proteins of unknown function: A pilot study with PH domains
Authors: Fidler David R. , Murphy Sarah E. , Courtis Katherine , Antonoudiou Pantelis , El‐Tohamy Rana , Ient Jonathan , Levine Timothy P.
Published: September 2016

Advances in membrane cell biology are hampered by the relatively high proportion of proteins with no known function. Such proteins are largely or entirely devoid of structurally significant domain annotations. Structural bioinformaticians have developed profile‐profile tools such as HHsearch (online version called HHpred), which can detect remote homologies that are missed by tools used to annotate databases. Here we have applied HHsearch to study a single structural fold in a single model organism as proof of principle. In the entire clan of protein domains sharing the pleckstrin homology domain fold in yeast, systematic application of HHsearch accurately identified known PH‐like domains. It also predicted 16 new domains in 13 yeast proteins many of which are implicated in intracellular traffic. One of these was Vps13p, where we confirmed the functional importance of the predicted PH‐like domain. Even though such predictions require considerable work to be corroborated, they are useful first steps. HHsearch should be applied more widely, particularly across entire proteomes of model organisms, to significantly improve database annotations.
Bilirubin levels and phototherapy use before and after neonatal red blood cell transfusions
Authors: Carroll Patrick D. , Christensen Robert D. , Baer Vickie L. , Sheffield Mark J. , Gerday Erick , Ilstrup Sarah J.
Published: September 2016

BACKGROUND

Our previous retrospective study suggested that red blood cell (RBC) transfusion of preterm neonates can be associated with an increase in bilirubin, but this has not been tested prospectively.

STUDY DESIGN AND METHODS

We studied neonates before and after RBC transfusions, recording serial bilirubin levels and whether they qualified for phototherapy. Because lysed RBCs release plasma‐free hemoglobin (Hb), a precursor to bilirubin, we also measured plasma free Hb and bilirubin from the donor blood.

RESULTS

We studied 50 transfusions given to 39 neonates. Gestation ages of transfused neonates, at birth, were 26 (24‐29) weeks (median [interquartile range]); birthweights were 750 (620‐1070) g. The study transfusion was given on Day of Life 9.9 (3.4‐19.2). In 20% (10/50) phototherapy was being administered at the beginning of and during the transfusion. In these patients neither the 4‐ to 6‐ nor the 24‐ to 36‐hour‐posttransfusion bilirubin levels were significantly higher than before transfusion. However, in 30% of the others (12/40) phototherapy was started (or restarted) after the transfusion and 15% had a posttransfusion bilirubin increase of at least 2.5 mg/dL. These neonates received donor blood with a higher plasma‐free Hb (p < 0.05).

CONCLUSIONS

Neonates commonly qualify for phototherapy after transfusion. A minority (15% in this series) have a posttransfusion bilirubin increase of at least 2.5 mg/dL. We speculate that neonates qualifying for a RBC transfusion, who are judged to be at high risk for bilirubin‐induced neurotoxicity, might benefit from checking their serum bilirubin level after the transfusion and providing donor blood with low plasma‐free Hb levels.
Profiling clinical platelet and plasma use to inform blood supply and contingency planning: PUPPY, the prospective utilization of platelets and plasma study
Authors: Fedele Pasquale L. , Polizzotto Mark N. , Grigoriadis George , Waters Neil , Comande Mary , Borosak Marija , Portbury David , Wood Erica M.
Published: September 2016

BACKGROUND

Demand for platelet (PLT) and plasma transfusions is increasing. Improved clinical supply and contingency planning requires greater understanding of usage profiles and urgency of clinical requirement.

STUDY DESIGN AND METHODS

This study was a random‐sample survey of PLT and plasma units produced in Victoria, Australia, to determine product disposition, recipient demographics, clinical indications for transfusion, and urgency (or “deferability”) of need. PLTs and fresh‐frozen plasma (FFP) were tagged with a case report form before distribution.

RESULTS

A total of 1252 PLT and 1837 FFP units were tagged, comprising 8.3 and 13.3% of all products issued during the study period. The fate of 1243 PLT and 1808 FFP units was determined. Of products issued, 72.2% of PLTs and 87.8% of FFP were transfused. Hematologic and oncologic disorders accounted for 63.9% of PLT transfusions, with acute myeloid leukemia alone accounting for 26%. Conversely, surgical patients received the largest proportion of FFP (40.4%), predominantly for cardiothoracic, solid organ transplant, and vascular surgery. Approximately 15% of PLT transfusions and 35% of plasma transfusions were required within 1 hour, and 80% of PLT transfusions and 90% of FFP transfusions were required within 24 hours. Wastage rates were higher in regional blood banks.

CONCLUSION

The PUPPY study is a comprehensive and detailed population‐based assessment of PLT and plasma usage, including urgency of use. It identifies specific clinical areas with high demand for PLT and FFP transfusion and demonstrates the high urgency of need for both products. These data inform clinical supply and contingency planning activities.
International validation of a dithiothreitol (DTT)‐based method to resolve the daratumumab interference with blood compatibility testing
Authors: Chapuy Claudia I. , Aguad Maria D. , Nicholson Rachel T. , AuBuchon James P. , Cohn Claudia S. , Delaney Meghan , Fung Mark K. , Unger Meredith , Doshi Parul , Murphy Michael F. , Dumont Larry J. , Kaufman Richard M. ,   the DARA‐DTT Study Group* for the BEST Collaborative
Published: September 2016

BACKGROUND

Daratumumab (DARA) consistently interferes with routine blood bank serologic testing by directly binding to CD38 expressed on reagent red blood cells (RBCs). Treating RBCs with dithiothreitol (DTT) eliminates the DARA interference. We conducted an international, multicenter, blinded study aimed at validating the DTT method for use by blood bank laboratories worldwide.

STUDY DESIGN AND METHODS

Paired plasma sample unknowns were sent to 25 participating blood bank laboratories. Sample 1 was spiked with DARA only (10 µg/mL), and Sample 2 with DARA plus a clinically significant RBC antibody (anti‐D [n = 6], anti‐Fya [n = 9], or anti‐s [n = 10]). Sites were instructed to perform an antibody screen with and without DTT‐treated RBCs and to use a DTT‐treated RBC panel for antibody identification. Qualitative data about the DTT method were collected by online survey. The primary outcome was the proportion of study sites able to identify the antibody unknown in the presence of DARA.

RESULTS

All sites observed the DARA interference with the antibody screen. The DARA interference was seen with all testing methods (gel, tube, or solid phase). Using the DTT method, 25 of 25 sites (100%) successfully identified the antibody unknown in the presence of DARA. Feedback on the DTT method was positive, with 17 of 19 (90%) sites responding to the survey indicating that they planned to use the DTT method to test clinical samples from DARA‐treated patients.

CONCLUSION

The DTT method is robust and reproducible and can be implemented by transfusion services worldwide to help provide safe blood products to patients treated with DARA.
Use of hydroxyethyl starch in leukocytapheresis procedures does not increase renal toxicity
Authors: Pagano Monica B. , Harmon Charles , Cooling Laura , Connelly‐Smith Laura , Mann Steven A. , Pham Huy P. , Marques Marisa B. , Schlueter Annette J. , Case Rosemary , King Karen E. , Cataife Guido , Wu Yanyun , Wong Edward C. C. , Winters Jeffrey L.
Published: September 2016

BACKGROUND

Hydroxyethyl starch (HES) is reportedly associated with an increased risk of renal failure and death when used for fluid resuscitation in critically ill patients. HES can be used during therapeutic leukocytapheresis (TL) procedures to enhance cell separation. The purpose of this study was to evaluate the occurrence of adverse events associated with HES during TL procedures.

STUDY DESIGN AND METHODS

We performed a retrospective review of patients who underwent TL with and without HES in the period 2009 to 2013 at six academic medical institutions.

RESULTS

A difference‐in‐difference regression analysis was used to estimate the mean change before and after TL in selected outcomes in the HES group relative to the average change in the non‐HES group. Selected outcomes included serum creatinine, estimated glomerular filtration rate (eGFR), and white blood cell (WBC) count. A total of 195 patients who underwent 278 TL procedures were studied. We found no significant differences in serum creatinine levels and eGFR on Days 1 and 7 after TL procedure between patients who received and those who did not receive HES. The rate of adverse events and overall and early mortality were similar in both groups. Patients with acute myeloid leukemia who received HES had greater WBC reduction when HES was used. Additionally, patients who received HES had improvement in pulmonary leukostasis symptoms.

CONCLUSION

HES, used at low doses during TL procedures, was not associated with adverse events previously ascribed to its use as a volume expander.
Coagulation status after therapeutic plasma exchange using citrate in kidney transplant recipients
Authors: Yamada Chisa , Pipe Steven W. , Zhao Lili , Leichtman Alan B. , Samaniego Milagros , Sung Randall S. , Davenport Robertson D.
Published: September 2016

BACKGROUND

Therapeutic plasma exchange (TPE) is increasingly used for treatment of antibody‐mediated rejection (AMR) after solid organ transplants. There is concern that TPE may increase risk of bleeding, although data are limited. After TPE, clot‐based coagulation tests may not accurately represent the levels of coagulation factors due to the effect of citrate. We investigated protein levels of fibrinogen using antigen detection method (FibAg) and correlated results with a clot‐based fibrinogen activity test (Fib).

STUDY DESIGN AND METHODS

Nine kidney transplant recipients who received TPE for AMR were investigated. Fib, FibAg, prothrombin time/international normalized ratio (PT/INR), partial thromboplastin time (PTT), coagulation factor X chromogenic activity (CFX), and ionized calcium (iCa) were measured at pre‐ and post‐TPE and 1, 3, 6, 9, 24, and 48 hours after the first TPE.

RESULTS

Mean Fib/FibAg ratio before TPE was 1.08; therefore, all Fib values were normalized (n) by dividing by 1.08. Overall, the mean normalized Fib (nFib)/FibAg ratio at post‐TPE was 0.89 and returned to close to 1.0 at 6 hours after the first TPE. Decreases in nFib, FibAg, and CFX and increases in PT/INR and PTT post‐TPE were observed. The lowest Fib, FibAg, CFX, platelet, and iCa levels were still at levels that would be considered sufficient for hemostasis at all time points.

CONCLUSION

The mean nFib/FibAg ratio after TPE was 0.89 and normalized in 6 hours, which demonstrates a persistent effect of citrate for up to 6 hours. Therefore, similar data observed in clot‐based tests of PT/INR and PTT may be falsely elevated up to 6 hours after TPE due to the citrate effect.
Probabilistic Modeling of Pitting Corrosion in Insulated Components Operating in Offshore Facilities
Authors: Shekari Elahe , Khan Faisal , Ahmed Salim
Published: September 2016

Pitting corrosion under insulation is one of the challenging issues for safe operation of offshore facilities. Degradation usually remains hidden causing the inspection of insulated assets to be equally challenging. The modeling of the pitting corrosion under insulation (CUI) helps us to better understand the current state of the asset and predict failure. This paper investigates the factors affecting the pit initiation and pit growth on equipment under insulation operating in offshore environments. A methodology is proposed for studying the pitting CUI characteristics, including pit initiation time, pit density, and maximum pit depth over time. The proposed methodology provides a practical and more effective asset life management approach when supported by inspection data. The practical application of the proposed methodology is demonstrated in this paper using a pressure vessel case study in an offshore platform.
Evaluating multipulse integration as a neural-health correlate in human cochlear-implant users: Relationship to spatial selectivity
Authors: Zhou Ning , Pfingst Bryan E.
Published: September 2016

The decrease of psychophysical detection thresholds as a function of pulse rate for a fixed-duration electrical pulse train is referred to as multipulse integration (MPI). The MPI slopes correlate with anatomical and physiological indices of cochlear health in guinea pigs with cochlear implants. The aim of the current study was to assess whether the MPI slopes were related to the spatial spread of activation by electrical stimulation. The hypothesis was that MPI is dependent on the total number of excitable neurons at the stimulation site, with broader neural excitation producing a steeper threshold decrease as a function of stimulation rate. MPI functions were measured at all stimulation sites in 22-site electrode arrays in human subjects. Some sites with steep MPI functions and other sites with shallow functions were assessed for spatial spread of excitation at 900 pps using a forward-masking paradigm. The results showed a correlation between the slopes of the forward-masking functions and the steepness of MPI, with broader stimulation predicting greater integration. The results are consistent with the idea that integration of multiple pulses in a pulse train relies on the number of excitable neurons at the stimulation site.
Heat shock protein–based therapy as a potential candidate for treating the sphingolipidoses
Authors: Kirkegaard Thomas , Gray James , Priestman David A. , Wallom Kerri-Lee , Atkins Jennifer , Olsen Ole Dines , Klein Alexander , Drndarski Svetlana , Petersen Nikolaj H. T. , Ingemann Linda , Smith David A. , Morris Lauren , Bornæs Claus , Jørgensen Signe Humle , Williams Ian , Hinsby Anders , Arenz Christoph , Begley David , Jäättelä Marja , Platt Frances M.
Published: September 2016

Increasing Hsp70 expression in lysosomes using the small-molecule arimoclomol ameliorates pathology in several animal models of sphingolipidoses.
Immunotherapy of non-Hodgkin’s lymphoma with a defined ratio of CD8
+
and CD4
+
CD19-specific chimeric antigen receptor–modified T cells
Authors: Turtle Cameron J. , Hanafi Laïla-Aïcha , Berger Carolina , Hudecek Michael , Pender Barbara , Robinson Emily , Hawkins Reed , Chaney Colette , Cherian Sindhu , Chen Xueyan , Soma Lorinda , Wood Brent , Li Daniel , Heimfeld Shelly , Riddell Stanley R. , Maloney David G.
Published: September 2016

A CD19 chimeric antigen receptor–modified T cell product with defined composition has potent antitumor activity.
Bugging cancer with guided swarming
Authors: Castro Cesar M.
Published: September 2016

Combining swarming of naturally occurring magneto-aerotactic bacteria with guided tumor targeting results in nanorobot carriers for chemotherapy payload delivery and potentially reduced side effects.
Can imaging predict future suicidal behavior?
Authors: Fung Lawrence K.
Published: September 2016

Brain concentrations of serotonin 1A receptor in the raphe nuclei determined by in vivo imaging may predict lethality of suicidal attempts.
Ex Vivo
Maturation Assay for Testing Antimalarial Sensitivity of Rodent Malaria Parasites
Authors: Chang Zi Wei , Malleret Benoit , Russell Bruce , Rénia Laurent , Claser Carla
Published: September 2016

Ex vivo
assay systems provide a powerful approach to studying human malaria parasite biology and to testing antimalarials. For rodent malaria parasites, short-term
in vitro
culture and
ex vivo
antimalarial susceptibility assays are relatively cumbersome, relying on
in vivo
passage for synchronization, since ring-stage parasites are an essential starting material. Here, we describe a new approach based on the enrichment of ring-stage
Plasmodium berghei
,
P. yoelii
, and
P. vinckei vinckei
using a single-step Percoll gradient. Importantly, we demonstrate that the enriched ring-stage parasites develop synchronously regardless of the parasite strain or species used. Using a flow cytometry assay with Hoechst and ethidium or MitoTracker dye, we show that parasite development is easily and rapidly monitored. Finally, we demonstrate that this approach can be used to screen antimalarial drugs.
Prospective Cohort Study of the Relative Abundance of Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli in the Gut of Patients Admitted to Hospitals
Authors: de Lastours Victoire , Chopin Dorothée , Jacquier Hervé , d'Humières Camille , Burdet Charles , Chau Françoise , Denamur Erick , Fantin Bruno
Published: September 2016

A total of 458 patients were prospectively included at hospital admission and screened for extended-spectrum-beta-lactamase-producing (ESBL)
Escherichia coli
carriage in 2007 and in 2010 to 2012. A 4-fold increase in ESBL carriage (3% to 12%), a 5-fold increase in numbers of community patients among ESBL carriers, and a higher number of multiple ESBL strains was found in the 2010 to 2012 period. ESBL
E. coli
represented the dominant
E. coli
strain (relative abundance, >50%) in 10/32 (31%) of ESBL carriers. This represents a major threat in terms of infectious risk and dissemination.
Genomic Characterization of Colistin Heteroresistance in Klebsiella pneumoniae during a Nosocomial Outbreak
Authors: Halaby Teysir , Kucukkose Emre , Janssen Axel B. , Rogers Malbert R. C. , Doorduijn Dennis J. , van der Zanden Adri G. M. , al Naiemi Nashwan , Vandenbroucke-Grauls Christina M. J. E. , van Schaik Willem
Published: September 2016

Klebsiella pneumoniae
is emerging as an important nosocomial pathogen due to its rapidly increasing multidrug resistance, which has led to a renewed interest in polymyxin antibiotics, such as colistin, as antibiotics of last resort. However, heteroresistance (i.e., the presence of a subpopulation of resistant bacteria in an otherwise susceptible culture) may hamper the effectiveness of colistin treatment in patients. In a previous study, we showed that colistin resistance among extended-spectrum-beta-lactamase (ESBL)-producing
K. pneumoniae
isolates emerged after the introduction of selective digestive tract decontamination (SDD) in an intensive care unit (ICU). In this study, we investigated heteroresistance to colistin among ESBL-producing
K. pneumoniae
isolates by using population analysis profiles (PAPs). We used whole-genome sequencing (WGS) to identify the mutations that were associated with the emergence of colistin resistance in these
K. pneumoniae
isolates. We found five heteroresistant subpopulations, with colistin MICs ranging from 8 to 64 mg/liter, which were derived from five clonally related, colistin-susceptible clinical isolates. WGS revealed the presence of mutations in the
lpxM
,
mgrB
,
phoQ
, and
yciM
genes in colistin-resistant
K. pneumoniae
isolates. In two strains,
mgrB
was inactivated by an IS
3
-like or IS
Kpn14
insertion sequence element. Complementation in
trans
with the wild-type
mgrB
gene resulted in these strains reverting to colistin susceptibility. The MICs for colistin-susceptible strains increased 2- to 4-fold in the presence of the mutated
phoQ
,
lpxM
, and
yciM
alleles. In conclusion, the present study indicates that heteroresistant
K. pneumoniae
subpopulations may be selected for upon exposure to colistin. Mutations in
mgrB
and
phoQ
have previously been associated with colistin resistance, but we provide experimental evidence for roles of mutations in the
yciM
and
lpxM
genes in the emergence of colistin resistance in
K. pneumoniae
.
Disruption of Membrane by Colistin Kills Uropathogenic Escherichia coli Persisters and Enhances Killing of Other Antibiotics
Authors: Cui Peng , Niu Hongxia , Shi Wanliang , Zhang Shuo , Zhang Hao , Margolick Joseph , Zhang Wenhong , Zhang Ying
Published: September 2016

Persisters are small populations of quiescent bacterial cells that survive exposure to bactericidal antibiotics and are responsible for many persistent infections and posttreatment relapses. However, little is known about how to effectively kill persister bacteria. In the work presented here, we found that colistin, a membrane-active antibiotic, was highly active against
Escherichia coli
persisters at high concentrations (25 or 50 μg/ml). At a clinically relevant lower concentration (10 μg/ml), colistin alone had no apparent effect on
E. coli
persisters. In combination with other drugs, this concentration of colistin enhanced the antipersister activity of gentamicin and ofloxacin but not that of ampicillin, nitrofurans, and sulfa drugs
in vitro
. The colistin enhancement effect was most likely due to increased uptake of the other antibiotics, as demonstrated by increased accumulation of fluorescence-labeled gentamicin. Interestingly, colistin significantly enhanced the activity of ofloxacin and nitrofurantoin but not that of gentamicin or sulfa drugs in the murine model of urinary tract infection. Our findings suggest that targeting bacterial membranes is a valuable approach to eradicating persisters and should have implications for more effective treatment of persistent bacterial infections.
Posaconazole Tablet Formulation at 400 Milligrams Daily Achieves Desired Minimum Serum Concentrations in Adult Patients with a Hematologic Malignancy or Stem Cell Transplant
Authors: Pham Aaron N. , Bubalo Joseph S. , Lewis James S.
Published: September 2016

We describe our experience using the posaconazole 400-mg delayed-release tablet formulation once daily in 20 patients with hematologic malignancy or hematopoietic stem cell transplant who were unable to attain prespecified target minimum serum (trough) concentrations for treatment or prophylaxis of invasive fungal infection. The higher dose allowed the majority of patients to achieve prespecified target trough concentrations without incurring additional toxicities.
Antileishmanial Activity of Ezetimibe: Inhibition of Sterol Biosynthesis,
In Vitro
Synergy with Azoles, and Efficacy in Experimental Cutaneous Leishmaniasis
Authors: Andrade-Neto Valter Viana , Cunha-Júnior Edézio Ferreira , Canto-Cavalheiro Marilene Marcuzzo do , Atella Geórgia Correa , Fernandes Talita de Almeida , Costa Paulo Roberto Ribeiro , Torres-Santos Eduardo Caio
Published: September 2016

Leishmaniasis affects mainly low-income populations in tropical regions. Radical innovation in drug discovery is time-consuming and expensive, imposing severe restrictions on the ability to launch new chemical entities for the treatment of neglected diseases. Drug repositioning is an attractive strategy for addressing a specific demand more easily. In this project, we have evaluated the antileishmanial activities of 30 drugs currently in clinical use for various morbidities. Ezetimibe, clinically used to reduce intestinal cholesterol absorption in dyslipidemic patients, killed
Leishmania amazonensis
promastigotes with a 50% inhibitory concentration (IC
50
) of 30 μM. Morphological analysis revealed that ezetimibe caused the parasites to become rounded, with multiple nuclei and flagella. Analysis by gas chromatography (GC)-mass spectrometry (MS) showed that promastigotes treated with ezetimibe had smaller amounts of C-14-demethylated sterols, and accumulated more cholesterol and lanosterol, than untreated promastigotes. We then evaluated the combination of ezetimibe with well-known antileishmanial azoles. The fractional inhibitory concentration index (FICI) indicated synergy when ezetimibe was combined with ketoconazole or miconazole. The activity of ezetimibe against intracellular amastigotes was confirmed, with an IC
50
of 20 μM, and ezetimibe reduced the IC
90
s of ketoconazole and miconazole from 11.3 and 11.5 μM to 4.14 and 8.25 μM, respectively. Subsequently, we confirmed the activity of ezetimibe
in vivo
, showing that it decreased lesion development and parasite loads in murine cutaneous leishmaniasis. We concluded that ezetimibe has promising antileishmanial activity and should be considered in combination with azoles in further preclinical and clinical studies.
Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation
Authors: Rizopoulos Zaira , Matuschewski Kai , Haussig Joana M.
Published: September 2016

Malarial parasites have evolved complex regulation of heme supply and disposal to adjust to heme-rich and -deprived host environments. In addition to its own pathway for heme biosynthesis,
Plasmodium
likely harbors mechanisms for heme scavenging from host erythrocytes. Elaborate compartmentalization of
de novo
heme synthesis into three subcellular locations, including the vestigial plastid organelle, indicates critical roles in life cycle progression. In this study, we systematically profile the essentiality of heme biosynthesis by targeted gene deletion of enzymes in early steps of this pathway. We show that disruption of endogenous heme biosynthesis leads to a first detectable defect in oocyst maturation and sporogony in the
Anopheles
vector, whereas blood stage propagation, colonization of mosquito midguts, or initiation of oocyst development occurs indistinguishably from that of wild-type parasites. Although sporozoites are produced by parasites lacking an intact pathway for heme biosynthesis, they are absent from mosquito salivary glands, indicative of a vital role for heme biosynthesis only in sporozoite maturation. Rescue of the first defect in sporogony permitted analysis of potential roles in liver stages. We show that liver stage parasites benefit from but do not strictly depend upon their own aminolevulinic acid synthase and that they can scavenge aminolevulinic acid from the host environment. Together, our experimental genetics analysis of
Plasmodium
enzymes for heme biosynthesis exemplifies remarkable shifts between the use of endogenous and host resources during life cycle progression.
The Adherent/Invasive Escherichia coli Strain LF82 Invades and Persists in Human Prostate Cell Line RWPE-1, Activating a Strong Inflammatory Response
Authors: Conte Maria P. , Aleandri Marta , Marazzato Massimiliano , Conte Antonietta L. , Ambrosi Cecilia , Nicoletti Mauro , Zagaglia Carlo , Gambara Guido , Palombi Fioretta , De Cesaris Paola , Ziparo Elio , Palamara Anna T. , Riccioli Anna , Longhi Catia
Published: September 2016

Adherent/invasive
Escherichia coli
(AIEC) strains have recently been receiving increased attention because they are more prevalent and persistent in the intestine of Crohn's disease (CD) patients than in healthy subjects. Since AIEC strains show a high percentage of similarity to extraintestinal pathogenic
E. coli
(ExPEC), neonatal meningitis-associated
E. coli
(NMEC), and uropathogenic
E. coli
(UPEC) strains, here we compared AIEC strain LF82 with a UPEC isolate (strain EC73) to assess whether LF82 would be able to infect prostate cells as an extraintestinal target. The virulence phenotypes of both strains were determined by using the RWPE-1 prostate cell line. The results obtained indicated that LF82 and EC73 are able to adhere to, invade, and survive within prostate epithelial cells. Invasion was confirmed by immunofluorescence and electron microscopy. Moreover, cytochalasin D and colchicine strongly inhibited bacterial uptake of both strains, indicating the involvement of actin microfilaments and microtubules in host cell invasion. Moreover, both strains belong to phylogenetic group B2 and are strong biofilm producers.
In silico
analysis reveals that LF82 shares with UPEC strains several virulence factors: namely, type 1 pili, the group II capsule, the vacuolating autotransporter toxin, four iron uptake systems, and the pathogenic island (PAI). Furthermore, compared to EC73, LF82 induces in RWPE-1 cells a marked increase of phosphorylation of mitogen-activated protein kinases (MAPKs) and of NF-κB already by 5 min postinfection, thus inducing a strong inflammatory response. Our
in vitro
data support the hypothesis that AIEC strains might play a role in prostatitis, and, by exploiting host-cell signaling pathways controlling the innate immune response, likely facilitate bacterial multiplication and dissemination within the male genitourinary tract.
Identification of a Novel
N
-Acetylmuramic Acid Transporter in Tannerella forsythia
Authors: Ruscitto Angela , Hottmann Isabel , Stafford Graham P. , Schäffer Christina , Mayer Christoph , Sharma Ashu
Published: September 2016

Tannerella forsythia
is a Gram-negative periodontal pathogen lacking the ability to undergo
de novo
synthesis of amino sugars
N
-acetylmuramic acid (MurNAc) and
N
-acetylglucosamine (GlcNAc) that form the disaccharide repeating unit of the peptidoglycan backbone.
T. forsythia
relies on the uptake of these sugars from the environment, which is so far unexplored. Here, we identified a novel transporter system of
T. forsythia
involved in the uptake of MurNAc across the inner membrane and characterized a homolog of the
Escherichia coli
MurQ etherase involved in the conversion of MurNAc-6-phosphate (MurNAc-6-P) to GlcNAc-6-P. The genes encoding these components were identified on a three-gene cluster spanning Tanf_08375 to Tanf_08385 located downstream from a putative peptidoglycan recycling locus. We show that the three genes, Tanf_08375, Tanf_08380, and Tanf_08385, encoding a MurNAc transporter, a putative sugar kinase, and a MurQ etherase, respectively, are transcriptionally linked. Complementation of the Tanf_08375 and Tanf_08380 genes together in
trans
, but not individually, rescued the inability of an
E. coli
mutant deficient in the phosphotransferase (PTS) system-dependent MurNAc transporter MurP as well as that of a double mutant deficient in MurP and components of the PTS system to grow on MurNAc. In addition, complementation with this two-gene construct in
E. coli
caused depletion of MurNAc in the medium, further confirming this observation. Our results show that the products of Tanf_08375 and Tanf_08380 constitute a novel non-PTS MurNAc transporter system that seems to be widespread among bacteria of the
Bacteroidetes
phylum. To the best of our knowledge, this is the first identification of a PTS-independent MurNAc transporter in bacteria.

IMPORTANCE
In this study, we report the identification of a novel transporter for peptidoglycan amino sugar
N
-acetylmuramic acid (MurNAc) in the periodontal pathogen
T. forsythia
. It has been known since the late 1980s that
T. forsythia
is a MurNAc auxotroph relying on environmental sources for this essential sugar. Most sugar transporters, and the MurNAc transporter MurP in particular, require a PTS phosphorelay to drive the uptake and concurrent phosphorylation of the sugar through the inner membrane in Gram-negative bacteria. Our study uncovered a novel type of PTS-independent MurNAc transporter, and although so far, it seems to be unique to
T. forsythia
, it may be present in a range of bacteria both of the oral cavity and gut, especially of the phylum
Bacteroidetes
.
Horizontal Transfer of the Salmonella enterica Serovar Infantis Resistance and Virulence Plasmid pESI to the Gut Microbiota of Warm-Blooded Hosts
Authors: Aviv Gili , Rahav Galia , Gal-Mor Ohad
Published: September 2016

Salmonella enterica
serovar Infantis is one of the prevalent salmonellae worldwide. Recently, we showed that the emergence of
S
. Infantis in Israel was facilitated by the acquisition of a unique megaplasmid (pESI) conferring multidrug resistance and increased virulence phenotypes. Here we elucidate the ecology, transmission properties, and regulation of pESI. We show that despite its large size (~280 kb), pESI does not impose a significant metabolic burden
in vitro
and that it has been recently fixed in the domestic
S
. Infantis population. pESI conjugation and the transcription of its pilus (
pil
) genes are inhibited at the ambient temperature (27°C) and by ≥1% bile but increased under temperatures of 37 to 41°C, oxidative stress, moderate osmolarity, and the microaerobic conditions characterizing the intestinal environment of warm-blooded animals. The pESI-encoded protein TraB and the oxygen homeostasis regulator Fnr were identified as transcriptional regulators of pESI conjugation. Using the mouse model, we show that following
S
. Infantis infection, pESI can be horizontally transferred to the gut microbiota, including to commensal
Escherichia coli
strains. Possible transfer, but not persistence, of pESI was also observed into Gram-positive mouse microbiota species, especially
Lactobacillus reuteri
. Moreover, pESI was demonstrated to further disseminate from gut microbiota to
S. enterica
serovar Typhimurium, in the context of gastrointestinal infection. These findings exhibit the ability of a selfish clinically relevant megaplasmid to distribute to and from the microbiota and suggest an overlooked role of the microbiota as a reservoir of mobile genetic elements and intermediator in the spread of resistance and virulence genes between commensals and pathogenic bacteria.

IMPORTANCE
Plasmid conjugation plays a key role in microbial evolution, enabling the acquisition of new phenotypes, including resistance and virulence.
Salmonella enterica
serovar Infantis is one of the ubiquitous salmonellae worldwide and a major cause of foodborne infections. Previously, we showed that the emergence of
S
. Infantis in Israel has involved the acquisition of a unique megaplasmid (pESI) conferring multidrug resistance and increased virulence phenotypes. Recently, the emergence of another
S
. Infantis strain carrying a pESI-like plasmid was identified in Italy, suggesting that the acquisition of pESI may be common to different emergent
S
. Infantis populations globally. Transmission of this plasmid to other strains or bacterial species is an alarming scenario. Understanding the ecology, regulation, and transmission properties of clinically relevant plasmids and the role of the microbiota in their spreading offers a new mechanism explaining the emergence of new pathogenic and resistant biotypes and may assist in the development of appropriate surveillance and prevention measures.
Individualized Responses of Gut Microbiota to Dietary Intervention Modeled in Humanized Mice
Authors: Smits Samuel A. , Marcobal Angela , Higginbottom Steven , Sonnenburg Justin L. , Kashyap Purna C.
Published: September 2016

Dietary modification has long been used empirically to modify symptoms in inflammatory bowel disease, irritable bowel syndrome, and a diverse group of diseases with gastrointestinal symptoms. There is both anecdotal and scientific evidence to suggest that individuals respond quite differently to similar dietary changes, and the highly individualized nature of the gut microbiota makes it a prime candidate for these differences. To overcome the typical confounding factors of human dietary interventions, here we employ ex-germfree mice colonized by microbiotas of three different humans to test how different microbiotas respond to a defined change in carbohydrate content of diet by measuring changes in microbiota composition and function using marker gene-based next-generation sequencing and metabolomics. Our findings suggest that the same diet has very different effects on each microbiota’s membership and function, which may in turn explain interindividual differences in response to a dietary ingredient.
Missed appendicitis: did unexpected intraluminal densities play a role?
Authors: Harper Rachel , Friedman Benjamin T , Strote Jared
Published: September 2016

A healthy 19-year-old boy presented to our emergency department with abdominal pain. His history, examination and laboratory evaluation raised concern for appendicitis. A CT study of the abdomen and pelvis was carried out by the radiologist and emergency physician and was notable only for a large amount of unexpected high-attenuation intraluminal material. With further history, this was thought to be most likely retained bismuth from over-the-counter medicine ingestion. The patient was discharged home without a diagnosis. Further review of the CT scan by a second radiologist revealed a concern for appendiceal enlargement and associated free fluid. The patient was called back for further evaluation and treatment and ultimately an appendectomy was performed. Physicians should be aware of the causes and impact of unexpected radiopaque intraluminal contents on radiological studies. Most commonly from ingested medicine, such findings can obscure mucosal details, mimic active bleeding or create a distraction from other abnormalities.