bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of platelet‐reactive alloantibodies and evaluation of cross‐match‐compatible platelets for the management of patients with transfusion refractoriness
Authors: Wang J. , Xia W. , Deng J. , Xu X. , Shao Y. , Ding H. , Chen Y. , Liu J. , Chen D. , Ye X. , Santoso S. 
Published: May 2017

SUMMARYBackgroundCross‐match‐compatible platelets can improve corrected count increments (CCIs) in alloimmunised patients with transfusion refractoriness. However, only a few studies mentioned that the specificities of platelet‐reactive alloantibodies can predict high reactivity in cross‐match assays among these patients.MethodsA total of 204 medical records of patients who were refractory to random single‐donor apheresis platelets between January 2014 and December 2014 were enrolled. Platelet‐reactive antibodies in patients' serum were screened by an enzyme‐linked immunosorbent assay (ELISA).The platelet cross‐match assays were performed by a solid‐phase adherence assay. The specificities of human leukocyte antigen (HLA) class I and human platelet antigens (HPAs) alloantibodies were determined by Luminex Single Antigen and Monoclonal Antibody‐specific Immobilization of Platelet Antigens (MAIPA) assays, respectively.ResultsAnti‐HLA and anti‐HPA alloantibodies were found in 114 of 204 (55.88%) patients, including 110 (96.49%) with anti‐HLA alloantibodies only, 2 (1.75%) with anti‐HPA alloantibodies (anti‐GPIIb/IIIa) only and 2 (1.75%) with both anti‐HLA and anti‐HPA alloantibodies (anti‐HPA‐3a and anti‐HPA‐5b). The most common HLA class I alloantibody phenotypes in cross‐match‐incompatible patients were HLA‐A23 (59.38%), ‐A24 (50.00%), ‐A02 (43.75%), ‐B27 (65.63%), ‐B40 (50.00%), ‐B18 (46.88%) and ‐B07 (43.75%). A total of 480 cross‐matched platelet units were administered in 82 of 114 alloimmunised patients with a mean CCI of 7800 ± 5200, a significant improvement over random platelet units (P < 0.001).ConclusionsNo development of additional platelet alloantibodies was observed during this platelet transfusion regiment. This study showed that transfusion of cross‐match‐compatible platelet units offers effective and safe management of platelet transfusion refractoriness (PTR). The finding of alloantibodies among cross‐match‐incompatible cases can be used as predictors for platelet donor selection.
Hepatic AA amyloidosis in a cat: cytologic and histologic identification of AA amyloid in macrophages
Authors: Neo‐suzuki Sakurako , Mineshige Takayuki , Kamiie Junichi , Kayanuma Hideki , Mochizuki Yukako , Hisasue Masaharu , Tsuchiya Ryo , Shirota Kinji 
Published: May 2017

AbstractA 3‐year‐old, spayed female, Domestic Shorthair cat presented with anorexia, lethargy, vomiting, probable hemoabdomen, and multiple masses on the right lateral liver lobe. Clinicopathologic and imaging abnormalities included anemia, azotemia, icterus, and hepatomegaly with hypoechoic masses. On cytologic evaluation of a fine‐needle aspiration of a liver mass there was abundant extracellular pink‐ to purple‐colored material between hepatocytes. The amorphous material was stained with direct fast scarlet (DFS), and green birefringent areas were observed under polarized light, confirming the presence of amyloid. A unique finding on the cytologic smear were macrophages containing amorphous and fibrillar amyloid‐like protein. Histopathologic examination using H&E and Congo red staining confirmed amyloid deposits within the space of Disse, along the sinusoids, portal tracts, blood vessel walls, and within the cytoplasm of macrophages. Immunohistochemical staining with anti‐AA amyloid antibodies further confirmed the presence of AA amyloid. To the author's knowledge, this is the first report of the cytologic finding of AA amyloid protein within macrophages and DFS stain detection of amyloid on a cytologic smear.
Implementing a fully automated high‐throughput strategy for blood donor genotyping
Authors: Dinardo C. L. , Bianchi J. V. , Dezan M. R. , Oliveira V. B. , Gomes F. C. A. , Gallucci A. J. O. , Levi J. E. , Krieger J. E. , Pereira A. C. , Santos H. C. , Domingues H. , Rocha V. G. , Mendrone‐Júnior A. , Sabino E. C. 
Published: May 2017

BackgroundHigh‐throughput molecular assays are essential tools for the search of rare blood donors.Aims and methodsTo evaluate a fully automated workflow designed for genotyping blood donors’ red blood cell (RBC) and platelets (PTL) antigens using the OpenArray Real‐time PCR system.Results5487 blood donors were genotyped using the proposed strategy in two steps: (1) nucleic acid purification in the QIASymphony® equipment and (2) genotyping of the most important RBC and PTL antigens by a customized assay designed for the OpenArray Real‐time PCR system. 142 662 single nucleotide polymorphisms were genotyped with 99·6% of accuracy in 15 work‐days. Software was created for the purpose of data interfacing and organization of a database to be used in a regular basis to find compatible blood for alloimmunized patients.ConclusionsThe proposed fully automated genotyping strategy is accurate, fast and suitable for meeting the needs of a laboratory of molecular immunohematology.
Metacarpophalangeal joint injury patterns on magnetic resonance imaging: A comparison in racing Standardbreds and Thoroughbreds
Authors: Olive Julien , Serraud Nicolas , Vila Thibault , Germain Jean‐Philippe 
Published: May 2017

AbstractThe metacarpal condyle has received extensive attention as a predominant site of overload arthropathy in racehorses. However, detailed descriptions of MRI lesion patterns for the metacarpophalangeal joint and comparisons between types of racing horses are currently lacking. Aims of this retrospective, cross‐sectional study were to describe and compare standing low‐field magnetic resonance findings in the metacarpophalangeal joints for groups of Standardbreds and Thoroughbreds using systematic semiquantitative scores. Data archives at the Clinique Vétérinaire Equine de Chantilly and Imavet were searched during the period from June 2010 to February 2016. Images were retrieved and reviewed by one reader. A total of 30 Standardbreds and 77 Thoroughbreds were sampled. The Thoroughbreds developed a much higher rate of stress fractures (40.3% vs. 10%) and overall more severe scores of subchondral sclerosis and bone marrow lesions than Standardbreds. The Standardbreds had more sites of subchondral bone defect and resorption, as well as higher synovitis and joint capsule thickening scores. Both Standardbreds and Thoroughbreds frequently developed multifocal subchondral lesions not limited to the distal palmar condyle. The Standardbreds had a higher tendency to develop dorsomedial joint pole subchondral injuries. Both Standardbreds and Thoroughbreds developed a noticeably high rate of periarticular soft tissue injuries (36.7% in Standardbreds and 20.8% in Thoroughbreds), which were rarely considered as the main contributors to the lameness. Findings supported the use of low‐field MRI as a whole‐organ assessment tool for Standardbreds and Thoroughbreds with suspected metacarpophalangeal joint injuries.
Identification of Novel Seroreactive Antigens in Johne's Disease Cattle by Using the Mycobacterium tuberculosis Protein Array
Authors: Bannantine John P. , Campo Joseph J. , Li Lingling , Randall Arlo , Pablo Jozelyn , Praul Craig A. , Raygoza Garay Juan Antonio , Stabel Judith R. , Kapur Vivek 
Published: May 2017

ABSTRACTJohne's disease, a chronic gastrointestinal inflammatory disease caused byMycobacterium aviumsubspeciesparatuberculosis, is endemic in dairy cattle and other ruminants worldwide and remains a challenge to diagnose using traditional serological methods. Given the close phylogenetic relationship betweenM. aviumsubsp.paratuberculosisand the human pathogenMycobacterium tuberculosis, here, we applied a whole-proteomeM. tuberculosisprotein array to identify seroreactive and diagnosticM. aviumsubsp.paratuberculosisantigens. A genome-scale pairwise analysis of amino acid identity levels between orthologous proteins inM. aviumsubsp.paratuberculosisandM. tuberculosisshowed an average of 62% identity, with more than half the orthologous proteins sharing >75% identity. Analysis of theM. tuberculosisprotein array probed with sera fromM. aviumsubsp.paratuberculosis-infected cattle showed antibody binding to 729M. tuberculosisproteins, with 58% of them having ≥70% identity toM. aviumsubsp.paratuberculosisorthologs. The results showed that only 4 of the top 40 seroreactiveM. tuberculosisantigens were orthologs of previously reportedM. aviumsubsp.paratuberculosisantigens, revealing the existence of a large number of previously unrecognized candidate diagnostic antigens. Enzyme-linked immunosorbent assay (ELISA) testing of 20M. aviumsubsp.paratuberculosisrecombinant proteins, representing reactive and nonreactiveM. tuberculosisorthologs, further confirmed that theM. tuberculosisarray has utility as a screening tool for identifying candidate antigens for Johne's disease diagnostics. Additional ELISA testing of field serum samples collected from dairy herds around the United States revealed that MAP2942c had the strongest seroreactivity with Johne's disease-positive samples. Collectively, our studies have considerably expanded the number of candidateM. aviumsubsp.paratuberculosisproteins with potential utility in the next generation of rationally designed Johne's disease diagnostic assays.
Development of a Novel Virus-Like Particle Vaccine Platform That Mimics the Immature Form of Alphavirus
Authors: Urakami Akane , Sakurai Atsuko , Ishikawa Momoko , Yap Moh Lan , Flores-Garcia Yevel , Haseda Yasunari , Aoshi Taiki , Zavala Fidel P. , Rossmann Michael G. , Kuno Sachiko , Ueno Ryuji , Akahata Wataru 
Published: May 2017

ABSTRACTVirus-like particles (VLPs) are noninfectious multiprotein structures that are engineered to self-assemble from viral structural proteins. Here, we developed a novel VLP-based vaccine platform utilizing VLPs from the chikungunya virus. We identified two regions within the envelope protein, a structural component of chikungunya, where foreign antigens can be inserted without compromising VLP structure. Our VLP displays 480 copious copies of an inserted antigen on the VLP surface in a highly symmetric manner and is thus capable of inducing strong immune responses against any inserted antigen. Furthermore, by mimicking the structure of the immature form of the virus, we altered our VLP'sin vivodynamics and enhanced its immunogenicity. We used the circumsporozoite protein (CSP) of thePlasmodium falciparummalaria parasite as an antigen and demonstrated that our VLP-based vaccine elicits strong immune responses against CSP in animals. The sera from immunized monkeys protected mice from malaria infection. Likewise, mice vaccinated withP. yoeliiCSP-containing VLPs were protected from an infectious sporozoite challenge. Hence, our uniquely engineered VLP platform can serve as a blueprint for the development of vaccines against other pathogens and diseases.
A Novel Stool PCR Test for Helicobacter pylori May Predict Clarithromycin Resistance and Eradication of Infection at a High Rate
Authors: Beckman Erin , Saracino Ilaria , Fiorini Giulia , Clark Courtney , Slepnev Vladimir , Patel Denise , Gomez Clarissa , Ponaka Reddy , Elagin Vecheslav , Vaira Dino 
Published: May 2017

ABSTRACT

Clarithromycin-based regimens are commonly used as a first-line therapy for
Helicobacter pylori
-positive patients; however, resistance to clarithromycin has led to treatment failures. The aim of this study was to evaluate the feasibility of using stool samples to detect the presence of
H. pylori
DNA while concurrently detecting mutations associated with resistance to clarithromycin. For this purpose, total DNA was extracted from 294 raw stool specimens from
H. pylori
-positive and -negative patients. TaqMan real-time PCR amplification was used to detect the presence of
H. pylori
as well as to predict the phenotype of the organism and the related outcome for patients treated with clarithromycin. Clarithromycin resistance was determined upon analysis of the PCR result. Patients were also tested by a urea breath test and were subjected to esophagogastroduodenoscopy, followed by histology, culture, and a rapid urease test, in order to obtain a consensus patient infection status. Of 294 total stool samples, 227 were deemed true positive. The sensitivity of
H. pylori
detection by PCR was 93.8%. Of 213 true-positive samples that were sequenced, 36.2% showed point mutations associated with clarithromycin resistance (A2142C, A2142G, A2143G). The final correlation of the mutant genotypes as determined by sequencing with the eradication of infection was 86%. We found that
Helicobacter pylori
DNA can be detected in human stool specimens with high sensitivity and can therefore be used to determine the presence of the bacterium without obtaining a biopsy sample. Moreover, genotypic resistance to clarithromycin can be predicted without obtaining a biopsy sample, facilitating the choice of the right therapeutic approach.
Outbreak of NDM-1-Producing Klebsiella pneumoniae in a Dutch Hospital, with Interspecies Transfer of the Resistance Plasmid and Unexpected Occurrence in Unrelated Health Care Centers
Authors: Bosch Thijs , Lutgens Suzanne P. M. , Hermans Mirjam H. A. , Wever Peter C. , Schneeberger Peter M. , Renders Nicole H. M. , Leenders Alexander C. A. P. , Kluytmans Jan A. J. W. , Schoffelen Annelot , Notermans Daan , Witteveen Sandra , Bathoorn Erik , Schouls Leo M. 
Published: May 2017

ABSTRACT

In the Netherlands, the number of cases of infection with New Delhi metallo-beta-lactamase (NDM)-positive
Enterobacteriaceae
is low. Here, we report an outbreak of NDM-1-producing
Klebsiella pneumoniae
infection in a Dutch hospital with interspecies transfer of the resistance plasmid and unexpected occurrence in other unrelated health care centers (HCCs). Next-generation sequencing was performed on 250 carbapenemase-producing
Enterobacteriaceae
isolates, including 42 NDM-positive isolates obtained from 29 persons at the outbreak site. Most outbreak isolates were
K. pneumoniae
(
n
= 26) and
Escherichia coli
(
n
= 11), but 5 isolates comprising three other
Enterobacteriaceae
species were also cultured. The 26
K. pneumoniae
isolates had sequence type 873 (ST873), as did 7 unrelated
K. pneumoniae
isolates originating from five geographically dispersed HCCs. The 33 ST873 isolates that clustered closely together using whole-genome multilocus sequence typing (wgMLST) carried the same plasmids and had limited differences in the resistome. The 11
E. coli
outbreak isolates showed great variety in STs, did not cluster using wgMLST, and showed considerable diversity in resistome and plasmid profiles. The
bla
NDM-1
gene-carrying plasmid present in the ST873
K. pneumoniae
isolates was found in all the other
Enterobacteriaceae
species cultured at the outbreak location and in a single
E. coli
isolate from another HCC. We describe a hospital outbreak with an NDM-1-producing
K. pneumoniae
strain from an unknown source that was also found in patients from five other Dutch HCCs in the same time frame without an epidemiological link. Interspecies transfer of the resistance plasmid was observed in other
Enterobacteriaceae
species isolated at the outbreak location and in another HCC.
Reactive Oxygen Species-Mediated c-Jun NH
2
-Terminal Kinase Activation Contributes to Hepatitis B Virus X Protein-Induced Autophagy via Regulation of the Beclin-1/Bcl-2 Interaction
Authors: Zhong Linmao , Shu Wangqin , Dai Wangbin , Gao Bo , Xiong Sidong 
Published: May 2017

ABSTRACT
Autophagy is closely associated with the regulation of hepatitis B virus (HBV) replication. HBV X protein (HBx), a multifunctional regulator in HBV-associated biological processes, has been demonstrated to be crucial for autophagy induction by HBV. However, the molecular mechanisms of autophagy induction by HBx, especially the signaling pathways involved, remain elusive. In the present investigation, we demonstrated that HBx induced autophagosome formation independently of the class I phosphatidylinositol 3-kinase (PI3K)/AKT/mTOR signaling pathway. In contrast, the class III PI3K(VPS34)/beclin-1 pathway was revealed to be critical for HBx-induced autophagosome formation. Further study showed that HBx did not affect the level of VPS34 and beclin-1 expression but inhibited beclin-1/Bcl-2 association, and c-Jun NH2-terminal kinase (JNK) signaling was found to be important for this process. Moreover, it was found that HBx treatment led to the generation of reactive oxygen species (ROS), and inhibition of ROS activity abrogated both JNK activation and autophagosome formation. Of importance, ROS-JNK signaling was also revealed to play an important role in HBV-induced autophagosome formation and subsequent HBV replication. These data may provide deeper insight into the mechanisms of autophagy induction by HBx and help in the design of new therapeutic strategies against HBV infection.

IMPORTANCE
HBx plays a key role in diverse HBV-associated biological processes, including autophagy induction. However, the molecular mechanisms of autophagy induction by HBx, especially the signaling pathways involved, remain elusive. In the present investigation, we found that HBx induced autophagy independently of the class I PI3K/AKT/mTOR signaling pathway, while the class III PI3K(VPS34)/beclin-1 pathway was revealed to be crucial for this process. Further data showed that ROS-JNK activation by HBx resulted in the release of beclin-1 from its association with Bcl-2 to form a complex with VPS34, thus enhancing autophagosome formation. Of importance, ROS-JNK signaling was also demonstrated to be critical for HBV replication via regulation of autophagy induction. These data help to elucidate the molecular mechanisms of autophagy induction by HBx/HBV and might be useful for designing novel therapeutic approaches to HBV infection.
Virus-Like Vesicles of Kaposi's Sarcoma-Associated Herpesvirus Activate Lytic Replication by Triggering Differentiation Signaling
Authors: Gong Danyang , Dai Xinghong , Xiao Yuchen , Du Yushen , Chapa Travis J. , Johnson Jeffrey R. , Li Xinmin , Krogan Nevan J. , Deng Hongyu , Wu Ting-Ting , Sun Ren 
Published: May 2017

ABSTRACT
Virus-like vesicles (VLVs) are membrane-enclosed vesicles that resemble native enveloped viruses in organization but lack the viral capsid and genome. During the productive infection of tumor-associated gammaherpesviruses, both virions and VLVs are produced and are released into the extracellular space. However, studies of gammaherpesvirus-associated VLVs have been largely restricted by the technical difficulty of separating VLVs from mature virions. Here we report a strategy of selectively isolating VLVs by using a Kaposi's sarcoma-associated herpesvirus (KSHV) mutant that is defective in small capsid protein and is unable to produce mature virions. Using mass spectrometry analysis, we found that VLVs contained viral glycoproteins required for cellular entry, as well as tegument proteins involved in regulating lytic replication, but lacked capsid proteins. Functional analysis showed that VLVs induced the expression of the viral lytic activator RTA, initiating KSHV lytic gene expression. Furthermore, employing RNA sequencing, we performed a genomewide analysis of cellular responses triggered by VLVs and found that PRDM1, a master regulator in cell differentiation, was significantly upregulated. In the context of KSHV replication, we demonstrated that VLV-induced upregulation of PRDM1 was necessary and sufficient to reactivate KSHV by activating its RTA promoter. In sum, our study systematically examined the composition of VLVs and demonstrated their biological roles in manipulating host cell responses and facilitating KSHV lytic replication.

IMPORTANCE
Cells lytically infected with tumor-associated herpesviruses produce a high proportion of virus-like vesicles (VLVs). The composition and function of VLVs have not been well defined, largely due to the inability to efficiently isolate VLVs that are free of virions. Using a cell system capable of establishing latent KSHV infection and robust reactivation, we successfully isolated VLVs from a KSHV mutant defective in the small capsid protein. We quantitatively analyzed proteins and microRNAs in VLVs and characterized the roles of VLVs in manipulating host cells and facilitating viral infection. More importantly, we demonstrated that by upregulating PRDM1 expression, VLVs triggered differentiation signaling in targeted cells and facilitated viral lytic infection via activation of the RTA promoter. Our study not only demonstrates a new strategy for isolating VLVs but also shows the important roles of KSHV-associated VLVs in intercellular communication and the viral life cycle.
Hepatitis C Virus Lipoviroparticles Assemble in the Endoplasmic Reticulum (ER) and Bud off from the ER to the Golgi Compartment in COPII Vesicles
Authors: Syed Gulam H. , Khan Mohsin , Yang Song , Siddiqui Aleem 
Published: May 2017

ABSTRACT

Hepatitis C virus (HCV) exists as a lipoprotein-virus hybrid lipoviroparticle (LVP).
In vitro
studies have demonstrated the importance of apolipoproteins in HCV secretion and infectivity, leading to the notion that HCV coopts the secretion of very-low-density lipoprotein (VLDL) for its egress. However, the mechanisms involved in virus particle assembly and egress are still elusive. The biogenesis of VLDL particles occurs in the endoplasmic reticulum (ER), followed by subsequent lipidation in the ER and Golgi compartment. The secretion of mature VLDL particles occurs through the Golgi secretory pathway. HCV virions are believed to latch onto or fuse with the nascent VLDL particle in either the ER or the Golgi compartment, resulting in the generation of LVPs. In our attempt to unravel the collaboration between HCV and VLDL secretion, we studied HCV particles budding from the ER
en route
to the Golgi compartment in COPII vesicles. Biophysical characterization of COPII vesicles fractionated on an iodixanol gradient revealed that HCV RNA is enriched in the highly buoyant COPII vesicle fractions and cofractionates with apolipoprotein B (ApoB), ApoE, and the HCV core and envelope proteins. Electron microscopy of immunogold-labeled microsections revealed that the HCV envelope and core proteins colocalize with apolipoproteins and HCV RNA in Sec31-coated COPII vesicles. Ultrastructural analysis also revealed the presence of HCV structural proteins, RNA, and apolipoproteins in the Golgi stacks. These findings support the hypothesis that HCV LVPs assemble in the ER and are transported to the Golgi compartment in COPII vesicles to embark on the Golgi secretory route.

IMPORTANCE
HCV assembly and release accompany the formation of LVPs that circulate in the sera of HCV patients and are also produced in an
in vitro
culture system. The pathway of HCV morphogenesis and secretion has not been fully understood. This study investigates the exact site where the association of HCV virions with host lipoproteins occurs. Using immunoprecipitation of COPII vesicles and immunogold electron microscopy (EM), we characterize the existence of LVPs that cofractionate with lipoproteins, viral proteins, RNA, and vesicular components. Our results show that this assembly occurs in the ER, and LVPs thus formed are carried through the Golgi network by vesicular transport. This work provides a unique insight into the HCV LVP assembly process within infected cells and offers opportunities for designing antiviral therapeutic cellular targets.