bluuebunny/crossref_metadata_2025_split huggingface dataset:
Acquired RhD mosaicism identifies fibrotic transformation of thrombopoietin receptor‐mutated essential thrombocythemia
Authors: Montemayor‐Garcia Celina , Coward Rebecca , Albitar Maher , Udani Rupa , Jain Prachi , Koklanaris Eleftheria , Battiwalla Minoo , Keel Siobán , Klein Harvey G. , Barrett A. John , Ito Sawa 
Published: June 2017

BACKGROUNDAcquired copy‐neutral loss of heterozygosity has been described in myeloid malignant progression with an otherwise normal karyotype.CASE REPORTA 65‐year‐old woman with MPL‐mutated essential thrombocythemia and progression to myelofibrosis was noted upon routine pretransplant testing to have mixed field reactivity with anti‐D and an historic discrepancy in RhD type. The patient had never received transfusions or transplantation.RESULTSGel immunoagglutination revealed group A red blood cells and a mixed‐field reaction for the D phenotype, with a predominant D‐negative population and a small subset of circulating red blood cells carrying the D antigen. Subsequent genomic microarray single nucleotide polymorphism profiling revealed copy‐neutral loss of heterozygosity of chromosome 1 p36.33‐p34.2, a known molecular mechanism underlying fibrotic progression of MPL‐mutated essential thrombocythemia. The chromosomal region affected by this copy‐neutral loss of heterozygosity encompassed the RHD, RHCE, and MPL genes. We propose a model of chronological molecular events that is supported by RHD zygosity assays in peripheral lymphoid and myeloid‐derived cells.CONCLUSIONCopy‐neutral loss of heterozygosity events that lead to clonal selection and myeloid malignant progression may also affect the expression of adjacent unrelated genes, including those encoding for blood group antigens. Detection of mixed‐field reactions and investigation of discrepant blood typing results are important for proper transfusion support of these patients and can provide useful surrogate markers of myeloproliferative disease progression.
Medical and economic implications of strategies to prevent alloimmunization in sickle cell disease
Authors: Gehrie Eric A. , Ness Paul M. , Bloch Evan M. , Kacker Seema , Tobian Aaron A. R. 
Published: June 2017

BACKGROUNDThe pathogenesis of alloimmunization is not well understood, and initiatives that aim to reduce the incidence of alloimmunization are generally expensive and either ineffective or unproven. In this review, we summarize the current medical literature regarding alloimmunization in the sickle cell disease (SCD) population, with a special focus on the financial implications of different approaches to prevent alloimmunization.STUDY DESIGN AND METHODSA review of EMBASE and MEDLINE data from January 2006 through January 2016 was conducted to identify articles relating to complications of SCD. The search was specifically designed to capture articles that evaluated the costs of various strategies to prevent alloimmunization and its sequelae.RESULTSCurrently, there is no proven, inexpensive way to prevent alloimmunization among individuals with SCD. Serologic matching programs are not uniformly successful in preventing alloimmunization, particularly to Rh antigens, because of the high frequency of variant Rh alleles in the SCD population. A genotypic matching program could offer some cost savings compared to a serologic matching program, but the efficacy of gene matching for the prevention of alloimmunization is largely unproven, and large‐scale implementation could be expensive.CONCLUSIONSFuture reductions in the costs associated with genotype matching could make a large‐scale program economically feasible. Novel techniques to identify patients at highest risk for alloimmunization could improve the cost effectiveness of antigen matching programs. A clinical trial comparing the efficacy of serologic matching to genotype matching would be informative.
Synthetic Cyclic Peptomers as Type III Secretion System Inhibitors
Authors: Lam Hanh , Schwochert Joshua , Lao Yongtong , Lau Tannia , Lloyd Cameron , Luu Justin , Kooner Olivia , Morgan Jessica , Lokey Scott , Auerbuch Victoria 
Published: June 2017

ABSTRACT

Antibiotic-resistant bacteria are an emerging threat to global public health. New classes of antibiotics and tools for antimicrobial discovery are urgently needed. Type III secretion systems (T3SS), which are required by dozens of Gram-negative bacteria for virulence but largely absent from nonpathogenic bacteria, are promising virulence blocker targets. The ability of mammalian cells to recognize the presence of a functional T3SS and trigger NF-κB activation provides a rapid and sensitive method for identifying chemical inhibitors of T3SS activity. In this study, we generated a HEK293 stable cell line expressing green fluorescent protein (GFP) driven by a promoter containing NF-κB enhancer elements to serve as a readout of T3SS function. We identified a family of synthetic cyclic peptide-peptoid hybrid molecules (peptomers) that exhibited dose-dependent inhibition of T3SS effector secretion in
Yersinia pseudotuberculosis
and
Pseudomonas aeruginosa
without affecting bacterial growth or motility. Among these inhibitors, EpD-3′N, EpD-1,2N, EpD-1,3′N, EpD-1,2,3′N, and EpD-1,2,4′N exhibited strong inhibitory effects on translocation of the
Yersinia
YopM effector protein into mammalian cells (>40% translocation inhibition at 7.5 μM) and showed no toxicity to mammalian cells at 240 μM. In addition, EpD-3′N and EpD-1,2,4′N reduced the rounding of HeLa cells caused by the activity of
Yersinia
effector proteins that target the actin cytoskeleton. In summary, we have discovered a family of novel cyclic peptomers that inhibit the injectisome T3SS but not the flagellar T3SS.
Preclinical Characterization of PC786, an Inhaled Small-Molecule Respiratory Syncytial Virus L Protein Polymerase Inhibitor
Authors: Coates Matthew , Brookes Daniel , Kim Young-In , Allen Heather , Fordyce Euan A. F. , Meals Elizabeth A. , Colley Thomas , Ciana Claire-Lise , Parra Guillaume F. , Sherbukhin Vladimir , Stockwell Jennifer A. , Thomas Jennifer C. , Hunt S. Fraser , Anderson-Dring Lauren , Onions Stuart T. , Cass Lindsey , Murray Peter J. , Ito Kazuhiro , Strong Pete , DeVincenzo John P. , Rapeport Garth 
Published: June 2017

ABSTRACT

Although respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infection in infants and young children, attempts to develop an effective therapy have so far proved unsuccessful. Here we report the preclinical profiles of PC786, a potent nonnucleoside RSV L protein polymerase inhibitor, designed for inhalation treatment of RSV infection. PC786 demonstrated a potent and selective antiviral activity against laboratory-adapted or clinical isolates of RSV-A (50% inhibitory concentration [IC
50
], <0.09 to 0.71 nM) and RSV-B (IC
50
, 1.3 to 50.6 nM), which were determined by inhibition of cytopathic effects in HEp-2 cells without causing detectable cytotoxicity. The underlying inhibition of virus replication was confirmed by PCR analysis. The effects of PC786 were largely unaffected by the multiplicity of infection (MOI) and were retained in the face of established RSV replication in a time-of-addition study. Persistent anti-RSV effects of PC786 were also demonstrated in human bronchial epithelial cells.
In vivo
intranasal once daily dosing with PC786 was able to reduce the virus load to undetectable levels in lung homogenates from RSV-infected mice and cotton rats. Treatment with escalating concentrations identified a dominant mutation in the L protein (Y1631H)
in vitro
. In addition, PC786 potently inhibited RSV RNA-dependent RNA polymerase (RdRp) activity in a cell-free enzyme assay and minigenome assay in HEp-2 cells (IC
50
, 2.1 and 0.5 nM, respectively). Thus, PC786 was shown to be a potent anti-RSV agent via inhibition of RdRp activity, making topical treatment with this compound a novel potential therapy for the treatment of human RSV infections.
The Nonantibiotic Macrolide EM703 Improves Survival in a Model of Quinolone-Treated Pseudomonas aeruginosa Airway Infection
Authors: Kasetty Gopinath , Bhongir Ravi K. V. , Papareddy Praveen , Herwald Heiko , Egesten Arne 
Published: June 2017

ABSTRACTMacrolide antibiotics are used as anti-inflammatory agents, e.g., for prevention of exacerbations in chronic obstructive pulmonary disease and cystic fibrosis. Several studies have shown improved outcomes after the addition of macrolides to β-lactam antibiotics for treatment of severe community-acquired pneumonia. However, a beneficial effect of macrolides in treating Gram-negative bacterial airway infections, e.g., those caused byPseudomonas aeruginosa, remains to be shown. Macrolide antibiotics have significant side effects, in particular, motility-stimulating activity in the gastrointestinal tract and promotion of bacterial resistance. In this study, EM703, a modified macrolide lacking antibiotic and motility-stimulating activities but with retained anti-inflammatory properties, was used as an adjunct treatment for experimentalP. aeruginosalung infection, in combination with a conventional antibiotic. Airway infections in BALB/cJRj mice were induced by nasal instillation ofP. aeruginosa; this was followed by treatment with the quinolone levofloxacin in the absence or presence of EM703. Survival, inflammatory responses, and cellular influx to the airways were monitored. Both pretreatment and simultaneous administration of EM703 dramatically improved survival in levofloxacin-treated mice withP. aeruginosaairway infections. In addition, EM703 reduced the levels of proinflammatory cytokines, increased the numbers of leukocytes in bronchoalveolar lavage fluid, and reduced the numbers of neutrophils present in lung tissue. In summary, the findings of this study show that the immunomodulatory properties of the modified macrolide EM703 can be important when treating Gram-negative pneumonia, as exemplified byP. aeruginosainfection in this study.
B10 Cells Alleviate Periodontal Bone Loss in Experimental Periodontitis
Authors: Wang Yuhua , Yu Xiaoqian , Lin Jiang , Hu Yang , Zhao Qian , Kawai Toshihisa , Taubman Martin A. , Han Xiaozhe 
Published: June 2017

ABSTRACT

B10 cells can regulate inflammatory responses in innate immunity. Toll-like receptors (TLRs) play an important role in B cell-mediated immune responses in periodontal disease. This study aimed to determine the effects of TLR-activated B10 cells on periodontal bone loss in experimental periodontitis. Spleen B cells isolated from C57BL/6J mice were cultured with
Porphyromonas gingivalis
lipopolysaccharide (LPS) and cytosine-phospho-guanine (CpG) oligodeoxynucleotides for 48 h. B10-enriched CD1d
hi
CD5
+
B cells were sorted by flow cytometry and were adoptively transferred to recipient mice through tail vein injection. At the same time,
P. gingivalis
-soaked ligatures were placed subgingivally around the maxillary second molars and remained there for 2 weeks before the mice were euthanized. Interleukin-10 (IL-10) production and the percentage of CD1d
hi
CD5
+
B cells were significantly increased with treatment with
P. gingivalis
LPS plus CpG compared to those in mice treated with
P. gingivalis
LPS or CpG alone. Mice with CD1d
hi
CD5
+
B cell transfer demonstrated reduced periodontal bone loss compared to the no-transfer group and the group with CD1d
lo
CD5
−
B cell transfer. Gingival IL-10 mRNA expression was significantly increased, whereas expressions of receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG), tumor necrosis factor alpha (TNF-α), and IL-1β were significantly inhibited in the CD1d
hi
CD5
+
B cell transfer group. The percentages of CD19
+
IL-10
+
cells, CD19
+
CD1d
hi
CD5
+
cells, and
P. gingivalis
-binding CD19
+
cells were significantly higher in recovered mononuclear cells from gingival tissues of the CD1d
hi
CD5
+
B cell transfer group than in tissues of the no-transfer group and the CD1d
lo
CD5
−
B cell transfer group. This study indicated that the adoptive transfer of B10 cells alleviated periodontal inflammation and bone loss in experimental periodontitis in mice.
Metabolism Dealing with Thermal Degradation of NAD
+
in the Hyperthermophilic Archaeon Thermococcus kodakarensis
Authors: Hachisuka Shin-ichi , Sato Takaaki , Atomi Haruyuki 
Published: June 2017

ABSTRACT

NAD
+
is an important cofactor for enzymatic oxidation reactions in all living organisms, including (hyper)thermophiles. However, NAD
+
is susceptible to thermal degradation at high temperatures. It can thus be expected that (hyper)thermophiles harbor mechanisms that maintain
in vivo
NAD
+
concentrations and possibly remove and/or reuse undesirable degradation products of NAD
+
. Here we confirmed that at 85°C, thermal degradation of NAD
+
results mostly in the generation of nicotinamide and ADP-ribose, the latter known to display toxicity by spontaneously linking to proteins. The hyperthermophilic archaeon
Thermococcus kodakarensis
possesses a putative ADP-ribose pyrophosphatase (ADPR-PPase) encoded by the TK2284 gene. ADPR-PPase hydrolyzes ADP-ribose to ribose 5-phosphate (R5P) and AMP. The purified recombinant TK2284 protein exhibited activity toward ADP-ribose as well as ADP-glucose. Kinetic analyses revealed a much higher catalytic efficiency toward ADP-ribose, suggesting that ADP-ribose was the physiological substrate. To gain insight into the physiological function of TK2284, a TK2284 gene disruption strain was constructed and examined. Incubation of NAD
+
in the cell extract of the mutant strain at 85°C resulted in higher ADP-ribose accumulation and lower AMP production compared with those in experiments with the host strain cell extract. The mutant strain also exhibited lower cell yield and specific growth rates in a synthetic amino acid medium compared with those of the host strain. The results obtained here suggest that the ADPR-PPase in
T. kodakarensis
is responsible for the cleavage of ADP-ribose to R5P and AMP, providing a means to utilize the otherwise dead-end product of NAD
+
breakdown.

IMPORTANCE
Hyperthermophilic microorganisms living under high temperature conditions should have mechanisms that deal with the degradation of thermolabile molecules. NAD
+
is an important cofactor for enzymatic oxidation reactions and is susceptible to thermal degradation to ADP-ribose and nicotinamide. Here we show that an ADP-ribose pyrophosphatase homolog from the hyperthermophilic archaeon
Thermococcus kodakarensis
converts the detrimental ADP-ribose to ribose 5-phosphate and AMP, compounds that can be directed to central carbon metabolism. This physiological role for ADP-ribose pyrophosphatases might be universal in hyperthermophiles, as their homologs are widely distributed among both hyperthermophilic bacteria and archaea.
The Proline Variant of the W[F/L/M][T/S]R Cyclic Di-GMP Binding Motif Suppresses Dependence on Signal Association for Regulator Function
Authors: Chodur Daniel M. , Guo Linda , Pu Meng , Bruger Eric , Fernandez Nico , Waters Chris , Rowe-Magnus Dean A. 
Published: June 2017

ABSTRACT

Vibrio vulnificus
is an estuarine bacterium and potent opportunistic human pathogen. It enters the food chain by asymptomatically colonizing a variety of marine organisms, most notably oysters. Expression of the
brp
-encoded extracellular polysaccharide, which enhances cell-surface adherence, is regulated by cyclic di-GMP (c-di-GMP) and the activator BrpT. The
Vibrio cholerae
and
Vibrio parahaemolyticus
homologs VpsT and CpsQ, directly bind c-di-GMP via a novel W[F/L/M][T/S]R motif, and c-di-GMP binding is absolutely required for activity. Notably, BrpT belongs to a distinct subclass of VpsT-like regulators that harbor a proline in the third position of the c-di-GMP binding motif (WLPR), and the impact of this change on activity is unknown. We show that the
brp
locus is organized as two linked operons with BrpT specifically binding to promoters upstream of
brpA
and
brpH
. Expression data and structural modeling suggested that BrpT might be less dependent on c-di-GMP binding for activity than VpsT or CpsQ. We show that the affinity of BrpT for c-di-GMP is low and that signal binding is not a requisite for BrpT function. Furthermore, a BrpT mutant engineered to carry a canonical WLTR motif (BrpT
P124T
) bound c-di-GMP with high affinity and its activity was now c-di-GMP dependent. Conversely, introduction of the WLPR motif into VpsT suppressed its dependence on c-di-GMP for activity. This is the first demonstration of reduced dependence on signal association for regulator function within this motif family. Thus, BrpT defines a new class of VpsT-like transcriptional regulators, and the WLPR motif variant may similarly liberate the activity of other subclass members.

IMPORTANCE
A
Vibrio
genome may encode nearly 100 proteins that make, break, and bind c-di-GMP, underscoring its central role in the physiology of these bacteria. The activity of the biofilm regulators VpsT of
V. cholerae
and CpsQ of
V. parahaemolyticus
is regulated by the direct binding of c-di-GMP via a novel W[F/L/M][T/S]R motif. The
V. vulnificus
homolog, BrpT, bears an unusual WLPR variant and remains active at low intracellular c-di-GMP levels. This suggests that the WLPR motif may also liberate the activity of other members of this subclass. A single point mutation at the 3rd position of the motif was sufficient to moderate dependence on c-di-GMP binding for activator function, highlighting the simplicity with which complex bacterial signaling networks can be rewired.
‘Milksops’ and ‘Bemedalled Old Men’: War Veterans and the War Youth Generation in the Weimar Republic
Authors: Mennen Kristian 
Published: June 2017

This article reconsiders traditional assumptions about the connection between the First World War and the rise of National Socialism in Germany, according to which politically radicalised war veterans joined the Freikorps after the war and formed the backbone of the Nazi membership and electorate. In questioning this view, the article first traces the political paths of actual veterans’ organisations. Whereas the largest veterans’ organisations were not politically active, the most distinctive ones – Reichsbanner and Stahlhelm – were not primarily responsible for a ‘brutalisation’ or radicalisation of Weimar political culture. Their definitions of ‘veteran’ and ‘front experience’ implicitly excluded the so-called ‘war youth generation’ from their narrative. Secondly, it is shown how representatives of this younger generation, lacking actual combat experience but moulded by war propaganda, determined the collective imagination of the First World War. The direct connection between the First World War and National Socialism can therefore primarily be found in the continuity of public and cultural imagination of war and of ‘war veterans’, and much less so in actual membership overlaps between veterans’ and Nazi movements.
Homofobia w polskiej kulturze wizualnej jako „nieodkryta” reakcja na przemiany norm seksualnych w Polsce po 1989 roku
Authors: Knut Paweł 
Published: June 2017

Homophobia in Polish visual culture as an “undiscovered” response to the transformation of sexual norms in Poland after 1989This article discusses the basic characteristics of various artistic practices aimed at stigmatizing nonheterosexuality that occurred in Poland after 1989 in response to a transformation of sexual norms and the rising emancipation of LGBT people. The form and content of selected practices is discussed in detail. Furthermore, the article argues that these practices can be classified as a form of so-called New National Art and signals the maladjustment of the language used by Polish art critique, which foreignizes and pathologizes this kind of art. The author also touches on the phenomenon of deliberate “not discovering” of these practices by national art institutions. Finally, he shows why the absence of artistic practices focused on exclusion and stigmatization of LGBT people seems to be beneficial for both major Polish art institutions and the “excluded” artists. Homofobia w polskiej kulturze wizualnej jako „nieodkryta” reakcja na przemiany norm seksualnych w Polsce po 1989 rokuArtykuł omawia podstawowe cechy praktyk artystycznych piętnujących nieheteroseksualność, które wystąpiły w Polsce po 1989 roku na skutek przemian norm seksualnych i zapoczątkowania procesu emancypacji osób LGBT. Charakteryzuje formy, jakie ta twórczość przyjmuje oraz treści, jakie wyraża. Wskazuje również na zasadność zaklasyfikowania tego rodzaju twórczości jako odmiany tzw. Nowej Sztuki Narodowej. Sygnalizuje problemy związane z używanym przez polską krytykę sztuki języka, który egzotyzuje i patologizuje tego rodzaju twórczość. Odnosi się również do zjawiska zamierzonego „nieodkrywania” tych praktyk artystycznych przez krajowe instytucje sztuki. Wreszcie, wskazuje dlaczego ta celowa nieobecność praktyki artystycznej nakierowanej na wykluczanie i piętnowanie osób LGBT wydaje się korzystna zarówno dla najważniejszych polskich instytucji sztuki, jak i dla „wykluczonych” twórców.
From assimilation, through transformation to studies within relationships: Understandings of nonheteronormative motherhood and ways forward
Authors: Kot Małgorzata 
Published: June 2017

From assimilation, through transformation to studies within relationships: Understandings of nonheteronormative motherhood and ways forwardNonheteronormative motherhood has been explored in many studies so far, aiming to explore the strategies of women who develop and maintain kinships in patriarchal settings. So far, the approaches to motherhood beyond hetero/normativity have been mostly focused on considering either assimilationist strategies or transformational potential of families of choice. In this article I would like to propose one of the possible ways enabling us to avoid these approaches. I argue that Rosi Braidotti’s framework of nomadic subjectivity and considering nonheteronormative motherhood as embodied experience might provide us with a lens through which we can look at mothering as de-centred, continuous becoming on a daily basis. Od asymilacji poprzez transformację do studiów w ramach relacji: rozumienia nieheteronormatywnego macierzyństwa i drogi na przyszłośćNieheteronormatywne macierzyństwo było do tej pory zgłębiane w wielu badaniach, mających na celu poznanie strategii kobiet, które tworzą i podtrzymują więzi w patriarchalnym otoczeniu. Dotychczas podejścia do macierzyństwa poza hetero/normatywnością były skupione przede wszystkim na rozważaniu asymilacyjnych strategii bądź transformatywnego potencjału rodzin z wyboru. W niniejszym artykule chciałabym zaproponować jedno z podejść umożliwiające uniknięcie wcześniejszych strategii. Dowodzę, że koncepcja nomadycznej tożsamości Rosi Braidotti oraz rozważanie nieheteronormatywnego macierzyństwa jako ucieleśnionego doświadczenia może zapewnić nam soczewkę, dzięki której możemy spojrzeć na macierzyństwo jako zdecentralizowany, ciągły proces codziennego stawania się matką.