bluuebunny/crossref_metadata_2025_split huggingface dataset:
Direct Neutralization of Type III Effector Translocation by the Variable Region of a Monoclonal Antibody to Yersinia pestis LcrV
Authors: Ivanov Maya I. , Hill Jim , Bliska James B. 
Published: March 2014

ABSTRACTPlague is an acute infection caused by the Gram-negative bacteriumYersinia pestis. Antibodies that are protective against plague target LcrV, an essential virulence protein and component of a type III secretion system ofY. pestis. Secreted LcrV localizes to the tips of type III needles on the bacterial surface, and its function is necessary for the translocation ofYersiniaouter proteins (Yops) into the cytosol of host cells infected byY. pestis. Translocated Yops counteract macrophage functions, for example, by inhibiting phagocytosis (YopE) or inducing cytotoxicity (YopJ). Although LcrV is the best-characterized protective antigen ofY. pestis, the mechanism of protection by anti-LcrV antibodies is not fully understood. Antibodies bind to LcrV at needle tips, neutralize Yop translocation, and promote opsonophagocytosis ofY. pestisby macrophagesin vitro. However, it is not clear if anti-LcrV antibodies neutralize Yop translocation directly or if they do so indirectly, by promoting opsonophagocytosis. To determine if the protective IgG1 monoclonal antibody (MAb) 7.3 is directly neutralizing, an IgG2a subclass variant, a deglycosylated variant, F(ab′)2, and Fab were tested for the ability to inhibit the translocation of Yops intoY. pestis-infected macrophagesin vitro. Macrophage cytotoxicity and cellular fractionation assays show that the Fc of MAb 7.3 is not required for the neutralization of YopJ or YopE translocation. In addition, the use of Fc receptor-deficient macrophages, and the use of cytochalasin D to inhibit actin polymerization, confirmed that opsonophagocytosis is not required for MAb 7.3 to neutralize translocation. These data indicate that the binding of the variable region of MAb 7.3 to LcrV is sufficient to directly neutralize Yop translocation.
Molecular Method Development and Establishment of a Database for Clinical and Epidemiological Herpes Simplex Virus 1 Strain Comparisons
Authors: Dean Amy B. , St. George Kirsten 
Published: March 2014

ABSTRACT
Previous methods of herpes simplex virus 1 (HSV-1) genotype analysis have lacked sufficient discriminatory power for strain analysis within genotypes. The hypervariable reiterative repeat regions in the US1 and US12 introns, known as ReIV, were targeted for strain comparison. PCR methods for these extremely GC-rich target regions were optimized to give reproducible amplicons that were visualized by capillary electrophoresis relative to size standards. Analysis of the size, shape, and pattern of the resulting signatures enabled strain discrimination. Primary clinical specimens were used to develop the assay and the analysis algorithm. A blinded clinical study of 147 in-state and 51 out-of-state samples, including matched specimen-isolate pairs, was then performed. All primary clinical samples had been collected between 2004 and 2011 for viral diagnosis and previously found to be positive for HSV-1 by real-time PCR. The combined database contained patterns from 264 samples collected from 199 patients with a total of 176 unique signatures, none of which were dominant in the population. Matches between the signatures of the more than 50 specimen-isolate pairs were always seen. Signatures also matched across multiple samples collected from individual patients (six such cases), as well as some additional signature matches where epidemiological links were likely. Results were reproducible on repeat testing of individual specimens, even after months in frozen storage. The protocol has multiple potential clinical and public health uses.
Cardiolipin Synthetase Is Involved in Antagonistic Interaction (Reverse CAMP Phenomenon) of Mycoplasma Species with Staphylococcus aureus Beta-Hemolysis
Authors: Kornspan Jonathan D. , Rottem Shlomo , Nir-Paz Ran 
Published: March 2014

ABSTRACTMycoplasma hyorhinishas been implicated in a variety of swine diseases. However, little is known about the hemolytic capabilities ofMycoplasmaspecies in general orM. hyorhinisin particular. In this study, we show thatM. hyorhinispossesses beta-hemolytic activity which may be involved in the invasion process.M. hyorhinisalso possesses antagonistic cooperativity (reverse CAMP phenomenon) withStaphylococcus aureusbeta-hemolysis, resulting in the protection of erythrocytes from the beta-hemolytic activity ofS. aureus(reverse CAMP). The reversed CAMP phenomenon has been attributed to phospholipase D (PLD) activity.In silicoanalysis of theM. hyorhinisgenome revealed the absence of thepldgene but the presence of theclsgene encoding cardiolipin synthetase, which contains two PLD active domains. The transformation ofMycoplasma gallisepticumthat has neither theclsgene nor the reverse CAMP phenomenon with theclsgene fromM. hyorhinisresulted in the reverse CAMP phenomenon, suggesting for the first time that reverse CAMP can be induced by cardiolipin synthetase.
Genotypes of Klebsiella oxytoca Isolates from Patients with Nosocomial Pneumonia Are Distinct from Those of Isolates from Patients with Antibiotic-Associated Hemorrhagic Colitis
Authors: Herzog Kathrin A. T. , Schneditz Georg , Leitner Eva , Feierl Gebhard , Hoffmann Karl Martin , Zollner-Schwetz Ines , Krause Robert , Gorkiewicz Gregor , Zechner Ellen L. , Högenauer Christoph 
Published: March 2014

ABSTRACT

Klebsiella oxytoca
acts as a pathobiont in the dysbiotic human intestinal microbiota, causing antibiotic-associated hemorrhagic colitis (AAHC), but it also infects other organs, resulting in pneumonia and urinary tract and skin infections. The virulence of
K. oxytoca
is still poorly understood. The production of a specific cytotoxin has been linked to AAHC pathogenesis. To investigate the clonal relationships of
K. oxytoca
with regard to clinical origin and virulence attributes, we established a multilocus sequence typing (MLST) method and analyzed 74 clinical
K. oxytoca
isolates from asymptomatic carriers and patients with AAHC, respiratory infections, and other infections. The isolates were phenotypically characterized, typed, and compared phylogenetically based on the sequences of seven housekeeping genes. MLST analysis yielded 60 sequence types, 12 of which were represented by more than one isolate. The phylogenetic tree distinguished clusters of
K. oxytoca
isolates between patients with AAHC and those with respiratory infections. Toxin-positive and -negative strains were observed within one sequence type. Our findings indicate that AAHC isolates share a genetic background. Interestingly,
K. oxytoca
isolates from nosocomial pneumonia showed a different genetic clustering, suggesting that these strains do not originate from the intestines or that they are specialized for respiratory tract colonization. Our results further indicate a polyphyletic origin and possible horizontal transfer of the genes involved in
K. oxytoca
cytotoxin production. This work provides evidence that
K. oxytoca
isolates colonizing the two main clinically relevant habitats (lower gastrointestinal [GI] tract and respiratory tract) of the human host are genetically distinct. Applications of this MLST analysis should help clarify the sources of nosocomial infections.
Highly Specific Salt Bridges Govern Bacteriophage P22 Icosahedral Capsid Assembly: Identification of the Site in Coat Protein Responsible for Interaction with Scaffolding Protein
Authors: Cortines Juliana R. , Motwani Tina , Vyas Aashay A. , Teschke Carolyn M. 
Published: March 2014

ABSTRACT

Icosahedral virus assembly requires a series of concerted and highly specific protein-protein interactions to produce a proper capsid. In bacteriophage P22, only coat protein (gp5) and scaffolding protein (gp8) are needed to assemble a procapsid-like particle, both
in vivo
and
in vitro
. In scaffolding protein's coat binding domain, residue R293 is required for procapsid assembly, while residue K296 is important but not essential. Here, we investigate the interaction of scaffolding protein with acidic residues in the N-arm of coat protein, since this interaction has been shown to be electrostatic. Through site-directed mutagenesis of genes 5 and 8, we show that changing coat protein N-arm residue 14 from aspartic acid to alanine causes a lethal phenotype. Coat protein residue D14 is shown by cross-linking to interact with scaffolding protein residue R293 and, thus, is intimately involved in proper procapsid assembly. To a lesser extent, coat protein N-arm residue E18 is also implicated in the interaction with scaffolding protein and is involved in capsid size determination, since a cysteine mutation at this site generated petite capsids. The final acidic residue in the N-arm that was tested, E15, is shown to only weakly interact with scaffolding protein's coat binding domain. This work supports growing evidence that surface charge density may be the driving force of virus capsid protein interactions.

IMPORTANCE
Bacteriophage P22 infects
Salmonella enterica
serovar Typhimurium and is a model for icosahedral viral capsid assembly. In this system, coat protein interacts with an internal scaffolding protein, triggering the assembly of an intermediate called a procapsid. Previously, we determined that there is a single amino acid in scaffolding protein required for P22 procapsid assembly, although others modulate affinity. Here, we identify partners in coat protein. We show experimentally that relatively weak interactions between coat and scaffolding proteins are capable of driving correctly shaped and sized procapsids and that the lack of these proper protein-protein interfaces leads to aberrant structures. The present work represents an important contribution supporting the hypothesis that virus capsid assembly is governed by seemingly simple interactions. The highly specific nature of the subunit interfaces suggests that these could be good targets for antivirals.
Depletion of the cdk Inhibitor p16
INK4a
Differentially Affects Proliferation of Established Cervical Carcinoma Cells
Authors: Pauck Alexander , Lener Barbara , Hoell Monika , Kaiser Andreas , Kaufmann Andreas M. , Zwerschke Werner , Jansen-Dürr Pidder 
Published: March 2014

ABSTRACT

Infections with high-risk human papillomaviruses (hrHPV) contribute to cervical carcinoma. The cdk inhibitor and tumor suppressor p16
INK4A
is consistently upregulated in cervical carcinoma cells for reasons that are poorly understood. We report here that downregulation of p16
INK4A
gene expression in three different cervical carcinoma cell lines reduced expression of the E7 oncogene, suggesting a positive feedback loop involving E7 and p16
INK4A
. p16
INK4A
depletion induced cellular senescence in HeLa but not CaSki and MS-751 cervical carcinoma cells.

IMPORTANCE
This study demonstrates that the cdk inhibitor p16
INK4A
, frequently used as surrogate marker for transforming infections by human papillomaviruses of the high-risk group, is required for high-level expression of the E7 oncoproteins of HPV-16, HPV-18, and HPV-45 in cervical carcinoma cells. It is also demonstrated that depletion of p16
INK4A
induces senescence in HeLa but not CaSki or MS-751 cervical carcinoma cells.
Mental disorders and the desire for death in patients receiving palliative care for cancer
Authors: Wilson Keith G , Dalgleish Tracy L , Chochinov Harvey Max , Chary Srini , Gagnon Pierre R , Macmillan Karen , De Luca Marina , O'Shea Fiona , Kuhl David , Fainsinger Robin L 
Published: March 2014

ObjectivesThe desire for death in terminally ill patients is associated with depression and anxiety, but not all patients who report it meet criteria for mental disorders. We examined the characteristics of subgroups of palliative cancer patients who expressed a desire for death that occurred either with or without a concurrent depressive or anxiety disorder.DesignCross-sectional survey.SettingEight Canadian palliative care programs.Participants377 patients with cancer.Main outcome measuresDesire for Death Rating Scale; Structured Interview of Symptoms and Concerns.ResultsMost participants (69.5%) had no desire for death. Of the remainder, 69 (18.3%) acknowledged occasional transient thoughts, and 46 (12.2%) reported an apparently genuine desire to die. In the latter group, 24 individuals (52.2%) were diagnosed with a mental disorder and 22 (44.8%) were not. Individuals with no serious desire for death and no mental disorder reported the least distress in physical, social, existential, and psychological symptoms and concerns; those with a mental disorder and a significant desire for death reported the most. The subgroup of patients with a serious desire for death but no concurrent mental disorders still reported increased distress due to physical symptoms and social concerns, as well as a higher prevalence of global suffering.ConclusionsThe expression of a desire for death by a terminally ill patient should raise a suspicion about mental health problems, but is not in itself clearly indicative of one. Nevertheless, it may serve as a catalyst to review the individual's physical symptom management and interpersonal concerns, and overall sense of suffering.
miR-30-5p Functions as a Tumor Suppressor and Novel Therapeutic Tool by Targeting the Oncogenic Wnt/β-Catenin/BCL9 Pathway
Authors: Zhao Jian-Jun , Lin Jianhong , Zhu Di , Wang Xujun , Brooks Daniel , Chen Ming , Chu Zhang-Bo , Takada Kohichi , Ciccarelli Bryan , Admin Samir , Tao Jianguo , Tai Yu-Tzu , Treon Steven , Pinkus Geraldine , Kuo Winston Patrick , Hideshima Teru , Bouxsein Mary , Munshi Nikhil , Anderson Kenneth , Carrasco Ruben 
Published: March 2014

Abstract
Wnt/β-catenin signaling underlies the pathogenesis of a broad range of human cancers, including the deadly plasma cell cancer multiple myeloma. In this study, we report that downregulation of the tumor suppressor microRNA miR-30-5p is a frequent pathogenetic event in multiple myeloma. Evidence was developed that miR-30-5p downregulation occurs as a result of interaction between multiple myeloma cells and bone marrow stromal cells, which in turn enhances expression of BCL9, a transcriptional coactivator of the Wnt signaling pathway known to promote multiple myeloma cell proliferation, survival, migration, drug resistance, and formation of multiple myeloma cancer stem cells. The potential for clinical translation of strategies to re-express miR-30-5p as a therapeutic approach was further encouraged by the capacity of miR-30c and miR-30 mix to reduce tumor burden and metastatic potential in vivo in three murine xenograft models of human multiple myeloma without adversely affecting associated bone disease. Together, our findings offer a preclinical rationale to explore miR-30-5p delivery as an effective therapeutic strategy to eradicate multiple myeloma cells in vivo. Cancer Res; 74(6); 1801–13. ©2014 AACR.
Id1-Induced IGF-II and Its Autocrine/Endocrine Promotion of Esophageal Cancer Progression and Chemoresistance—Implications for IGF-II and IGF-IR–Targeted Therapy
Authors: Li Bin , Tsao Sai Wah , Chan Kwok Wah , Ludwig Dale L. , Novosyadlyy Ruslan , Li Yuk Yin , He Qing Yu , Cheung Annie L.M. 
Published: March 2014

Abstract
Purpose: To investigate the autocrine/endocrine role of Id1-induced insulin-like growth factor-II (IGF-II) in esophageal cancer, and evaluate the potential of IGF-II- and IGF-type I receptor (IGF-IR)-targeted therapies.
Experimental Design: Antibody array-based screening was used to identify differentially secreted growth factors from Id1-overexpressing esophageal cancer cells. In vitro and in vivo assays were performed to confirm the induction of IGF-II by Id1, and to study the autocrine and endocrine effects of IGF-II in promoting esophageal cancer progression. Human esophageal cancer tissue microarray was analyzed for overexpression of IGF-II and its correlation with that of Id1 and phosphorylated AKT (p-AKT). The efficacy of intratumorally injected IGF-II antibody and intraperitoneally injected cixutumumab (fully human monoclonal IGF-IR antibody) was evaluated using in vivo tumor xenograft and experimental metastasis models.
Results: Id1 overexpression induced IGF-II secretion, which promoted cancer cell proliferation, survival, and invasion by activating AKT in an autocrine manner. Overexpression of IGF-II was found in 21 of 35 (60%) esophageal cancer tissues and was associated with upregulation of Id1 and p-AKT. IGF-II secreted by Id1-overexpressing esophageal cancer xenograft could instigate the growth of distant esophageal tumors, as well as promote metastasis of circulating cancer cells. Targeting IGF-II and IGF-IR had significant suppressive effects on tumor growth and metastasis in mice. Cixutumumab treatment enhanced the chemosensitivity of tumor xenografts to fluorouracil and cisplatin.
Conclusions: The Id1–IGF-II–IGF-IR–AKT signaling cascade plays an important role in esophageal cancer progression. Blockade of IGF-II/IGF-IR signaling has therapeutic potential in the management of esophageal cancer. Clin Cancer Res; 20(10); 2651–62. ©2014 AACR.
von Willebrand disease type 2A phenotypes IIC, IID and IIE: A day in the life of shear-stressed mutant von Willebrand factor
Authors: Huck Volker , Aponte-Santamaría Camilo , Obser Tobias , Grässle Sandra , Oyen Florian , Budde Ulrich , Schneppenheim Sonja , Baldauf Carsten , Gräter Frauke , Schneider Stefan W. , Schneppenheim Reinhard , Brehm Maria A. 
Published: March 2014

SummaryThe bleeding disorder von Willebrand disease (VWD) is caused by mutations of von Willebrand factor (VWF), a multimeric glycoprotein essential for platelet-dependent primary haemostasis. VWD type 2A–associated mutations each disrupt VWF biosynthesis and function at different stages, depending on the VWF domain altered by the mutation. These effects cause considerable heterogeneity in phenotypes and symptoms. To characterise the molecular mechanisms underlying the specific VWF deficiencies in VWD 2A/IIC, IID and IIE, we investigated VWF variants with patient-derived mutations either in the VWF pro-peptide or in domains D3 or CK. Additionally to static assays and molecular dynamics (MD) simulations we used microfluidic approaches to perform a detailed investigation of the shear-dependent function of VWD 2A mutants. For each group, we found distinct characteristics in their intracellular localisation visualising specific defects in biosynthesis which are correlated to respective multimer patterns. Using microfluidic assays we further determined shear flow-dependent characteristics in polymer-platelet-aggregate formation, platelet binding and string formation for all mutants. The phenotypes observed under flow conditions were not related to the mutated VWF domain. By MD simulations we further investigated how VWD 2A/IID mutations might alter the ability of VWF to form carboxy-terminal dimers. In conclusion, our study offers a comprehensive picture of shear-dependent and shear-independent dysfunction of VWD type 2A mutants. Furthermore, our microfluidic assay might open new possibilities for diagnosis of new VWD phenotypes and treatment choice for VWD patients with shear-dependent VWF dysfunctions that are currently not detectable by static tests.
Experimentally Increasing Titin Compliance in a Novel Mouse Model Attenuates the Frank-Starling Mechanism But Has a Beneficial Effect on Diastole
Authors: Methawasin Mei , Hutchinson Kirk R. , Lee Eun-Jeong , Smith John E. , Saripalli Chandra , Hidalgo Carlos G. , Ottenheijm Coen A.C. , Granzier Henk 
Published: March 2014

Background—
Experimentally upregulating compliant titins has been suggested as a therapeutic for lowering pathological diastolic stiffness levels. However, how increasing titin compliance impacts global cardiac function requires in-depth study. We investigate the effect of upregulating compliant titins in a novel mouse model with a genetically altered titin splicing factor; integrative approaches were used from intact cardiomyocyte mechanics to pressure-volume analysis and Doppler echocardiography.

Methods and Results—

Compliant titins were upregulated through deletion of the RNA Recognition Motif of the splicing factor RBM20 (
Rbm20

ΔRRM

mice). A genome-wide exon expression analysis and a candidate approach revealed that the phenotype is likely to be dominated by greatly increased lengths of titin’s spring elements. At both cardiomyocyte and left ventricular chamber levels, diastolic stiffness was reduced in heterozygous (+/−)
Rbm20

ΔRRM

mice with a further reduction in homozygous (−/−) mice at only the intact myocyte level. Fibrosis was present in only −/−
Rbm20

ΔRRM

hearts. The Frank-Starling Mechanism was reduced in a graded fashion in
Rbm20

ΔRRM

mice, at both the cardiomyocyte and left ventricular chamber levels. Exercise tests revealed an increase in exercise capacity in +/− mice.

Conclusions—
Titin is not only important in diastolic but also in systolic cardiac function. Upregulating compliant titins reduces diastolic chamber stiffness owing to the increased compliance of myocytes, but it depresses end-systolic elastance; under conditions of exercise, the beneficial effects on diastolic function dominate. Therapeutic manipulation of the RBM20-based splicing system might be able to minimize effects on fibrosis and systolic function while improving the diastolic function in patients with heart failure.
Focal bone lesions in hiv-positive patient treated with tenofovir
Authors: Mangioni Davide , Bandera Alessandra , Muscatello Antonio , Squillace Nicola , Crivellaro Cinzia , Guerra Luca , Messa Cristina , Gori Andrea 
Published: March 2014

Abstract

Background
Tenofovir is a widely used antiviral drug for the treatment of HIV and HBV infection. Although its side effects on renal function and bone metabolism are well known, there are no reports on focal bone lesions caused by this drug. Our case suggests this new, unusual but important scenario.

Case presentation
We report on a 46-year-old HIV-positive man treated with an antiretroviral regimen containing tenofovir who suddenly developed localized inflammatory bone lesions. The examinations performed ruled out all the disorders commonly associated with this clinical pattern, and the patient’s conditions improved only after the suspension of tenofovir.

Conclusions
The case study suggests a rare but severe adverse event, which should be taken into account when physicians treat HIV-positive patients with focal inflammatory bone lesions