bluuebunny/crossref_metadata_2025_split huggingface dataset:
Aggregating Phenotype in
Lactobacillus crispatus
Determines Intestinal Colonization and TLR2 and TLR4 Modulation in Murine Colonic Mucosa
Authors: Voltan Sandra , Castagliuolo Ignazio , Elli Marina , Longo Stefano , Brun Paola , D'Incà Renata , Porzionato Andrea , Macchi Veronica , Palù Giorgio , Sturniolo Giacomo C. , Morelli Lorenzo , Martines Diego 
Published: July 2007

ABSTRACT

The colonic microbiota is a major modulator of the mucosal immune system; therefore, its manipulation through supplementation with probiotics may significantly affect the host's immune responses. Since different probiotics seem to exert various effects in vivo, we tested the relevance of the autoaggregation phenotype on the intestinal persistence of lactobacilli and their ability to modulate the host's innate immune responses. After 14 days of diet supplementation, the aggregating strain
Lactobacillus crispatus
M247 but not aggregation-deficient isogenic mutant MU5 was recovered from the feces and colonic mucosa of mice. This observation was confirmed by strain-specific PCR amplification and by
Lactobacillus
-specific denaturing gradient gel electrophoresis analysis. Indeed,
L. crispatus
M247 increased Toll-like receptor 2 (TLR2) mRNA levels, while it reduced TLR4 mRNA and protein levels in the colonic mucosa, whereas MU5 was ineffective. In colonic epithelial cells (CMT-93 cells)
L. crispatus
M247 but not MU5 induced time-dependent extracellular signal-regulated kinase-1 (ERK1) tyrosine phosphorylation and TLR modulation, which were abolished in the presence of PD98059 (an ERK1 inhibitor). To assess the functional relevance of probiotic-induced TLR modulation, we determined the consequences of
L. crispatus
preexposure on TLR4 (lipopolysaccharide [LPS]) and TLR2 [Pam
3
Cys-Ser-(Lys)
4
] ligand-mediated effects in intestinal epithelial cells. Preexposure to
L. crispatus
M247 blunted LPS-induced interleukin-6 (IL-6) release and inhibition of CMT-93 migration over a wound edge, whereas it enhanced TLR2-mediated IL-10 up-regulation. In summary, the aggregation phenotype is required for
L. crispatus
persistence in the colon and for modulation of TLR2/TLR4 expression through an ERK-dependent pathway. We speculate that the aggregation phenotype in
L. crispatus
M247 is required to temper epithelial cell responsiveness to bacterial endotoxins, which thus affects the evolution of intestinal inflammatory processes.
Immunogenicity and Safety of a Combination of Two Serogroup B Meningococcal Outer Membrane Vesicle Vaccines
Authors: Sandbu Synne , Feiring Berit , Oster Philipp , Helland Oddveig S. , Bakke Hilde S. W. , Næss Lisbeth M. , Aase Audun , Aaberge Ingeborg S. , Kristoffersen Anne-Cathrine , Rydland Kjersti M. , Tilman Sandrine , Nøkleby Hanne , Rosenqvist Einar 
Published: July 2007

ABSTRACT
MenBvac and MeNZB are safe and efficacious vaccines against serogroup B meningococcal disease. MenBvac is prepared from a B:15:P1.7,16 meningococcal strain (strain 44/76), and MeNZB is prepared from a B:4:P1.7-2,4 strain (strain NZ98/254). At 6-week intervals, healthy adults received three doses of MenBvac (25 μg), MeNZB (25 μg), or the MenBvac and MeNZB (doses of 12.5 μg of each vaccine) vaccines combined, followed by a booster 1 year later. Two-thirds of the subjects who received a monovalent vaccine in the primary schedule received the other monovalent vaccine as a booster dose. The immune responses to the combined vaccine were of the same magnitude as the homologous responses to each individual vaccine observed. At 6 weeks after the third dose, 77% and 87% of the subjects in the combined vaccine group achieved serum bactericidal titers of ≥4 against strains 44/76 and NZ98/254, respectively, and 97% and 93% of the subjects achieved a fourfold or greater increase in opsonophagocytic activity against strains 44/76 and NZ98/254, respectively. For both strains, a trend of higher responses after the booster dose was observed in all groups receiving at least one dose of the respective strain-specific vaccine. Local and systemic reactions were common in all vaccine groups. Most reactions were mild or moderate in intensity, and there were no vaccine-related serious adverse events. The safety profile of the combined vaccine was not different from those of the separate monovalent vaccines. In conclusion, use of either of the single vaccines or the combination of MenBvac and MeNZB may have a considerable impact on the serogroup B meningococcal disease situation in many countries.
Immunization of Broiler Chickens against
Clostridium perfringens
-Induced Necrotic Enteritis
Authors: Kulkarni R. R. , Parreira V. R. , Sharif S. , Prescott J. F. 
Published: July 2007

ABSTRACT

Necrotic enteritis (NE) in broiler chickens is caused by
Clostridium perfringens
. Currently, no vaccine against NE is available and immunity to NE is not well characterized. Our previous studies showed that immunity to NE followed oral infection by virulent rather than avirulent
C. perfringens
strains and identified immunogenic secreted proteins apparently uniquely produced by virulent
C. perfringens
isolates. These proteins were alpha-toxin, glyceraldehyde-3-phosphate dehydrogenase, pyruvate:ferredoxin oxidoreductase (PFOR), fructose 1,6-biphosphate aldolase, and a hypothetical protein (HP). The current study investigated the role of each of these proteins in conferring protection to broiler chickens against oral infection challenges of different severities with virulent
C. perfringens
. The genes encoding these proteins were cloned and purified as histidine-tagged recombinant proteins from
Escherichia coli
and were used to immunize broiler chickens intramuscularly. Serum and intestinal antibody responses were assessed by enzyme-linked immunosorbent assay. All proteins significantly protected broiler chickens against a relatively mild challenge. In addition, immunization with alpha-toxin, HP, and PFOR also offered significant protection against a more severe challenge. When the birds were primed with alpha-toxoid and boosted with active toxin, birds immunized with alpha-toxin were provided with the greatest protection against a severe challenge. The serum and intestinal washings from protected birds had high antigen-specific antibody titers. Thus, we conclude that there are certain secreted proteins, in addition to alpha-toxin, that are involved in immunity to NE in broiler chickens.
The Binax NOW Test as a Tool for Diagnosis of Severe Acute Otitis Media and Associated Complications
Authors: Gisselsson-Solén Marie , Bylander Anita , Wilhelmsson Christina , Hermansson Ann , Melhus Åsa 
Published: July 2007

ABSTRACT

The diagnosis of acute otitis media (AOM) is often difficult, depending heavily on the experience and skills of the examiner. However, it is important to identify episodes of AOM that involve the risk of complications and to treat these episodes appropriately. The present study was performed in order to evaluate the use of a rapid antigen assay for
Streptococcus pneumoniae
, the Binax NOW test, as a diagnostic tool in patients with severe AOM and associated complications. The study included 70 patients with 74 episodes of AOM, 18 of them with complications. Cultures, Binax NOW tests, and a PCR assay were performed on nasopharyngeal secretions, middle ear fluid, and in some cases mastoid bone, cerebrospinal fluid, and urine. According to culture and PCR of the middle ear fluid, 30 (41%) of the episodes were caused by
S. pneumoniae
. The Binax NOW test was positive in 24 of these episodes (80%). It identified pneumococcal AOM independent of antibiotic treatment, and it was easily adapted to bone tissue. The test yielded sensitivity, specificity, and positive and negative predictive values for middle ear specimens of 85%, 100%, 100%, and 89%, respectively. The corresponding positive and negative values for predicting the bacterial etiology with nasopharyngeal secretions were 51% and 75%. This study showed that the Binax NOW test is a useful diagnostic tool for patients with severe AOM with or without complications.
Molecular Characterization of Vancomycin-Resistant
Enterococcus faecium
Isolates from Mainland China
Authors: Zheng Bo , Tomita Haruyoshi , Xiao Yong Hong , Wang Shan , Li Yun , Ike Yasuyoshi 
Published: July 2007

ABSTRACT

Little is known about vancomycin-resistant enterococci in China. Thirteen pulsed-field gel electrophoresis-confirmed heterogeneous VanA-type vancomycin-resistant
Enterococcus faecium
(VRE) isolates were obtained from five Chinese hospitals from 2001 to 2005. The isolates were typed by multilocus sequence typing into nine different sequence types (STs), including five new STs (ST18, ST25, ST78, ST203, ST320, ST321, ST322, ST323, and ST335). Vancomycin resistance in each isolate was encoded on conjugative plasmids; two of the plasmids, pZB18 (67 kbp) and pZB22 (200 kbp), were highly conjugative and were able to transfer at high frequencies of around 10
−4
and 10
−7
per donor cell in broth mating, respectively. None of the plasmids identified in these isolates carried
traA
, which is usually conserved in the pMG1-like highly conjugative plasmid for
E. faecium
, implying that pZB18 and pZB22 were novel types of a highly conjugative plasmid in enterococci. Thirteen Tn
1546
-like elements encoding VanA-type VRE on the conjugative plasmids were classified into six types (types I to VI), and most of them contained both IS
1216V
and IS
1542
insertions. The isolates carrying the type II element were predominant. The six type elements were different from that of a VanA-type
Enterococcus faecalis
strain isolated from Chinese chicken meat. The results suggested that the disseminations of VRE in these areas were by Tn
1546
-like elements being acquired by the conjugative plasmids and transferred among
E. faecium
strains.
Impact of the COBAS AmpliPrep/COBAS AMPLICOR HIV-1 MONITOR Test, Version 1.5, on Clinical Laboratory Operations
Authors: Germer Jeffrey J. , Bendel Jordan L. , Dolenc Craig A. , Nelson Sarah R. , Masters Amanda L. , Gerads Tara M. , Mandrekar Jayawant N. , Mitchell P. Shawn , Yao Joseph D. C. 
Published: July 2007

ABSTRACT
The COBAS AmpliPrep/COBAS AMPLICOR HIV-1 MONITOR Test, version 1.5 (CAP/CA), and the COBAS AMPLICOR HIV-1 MONITOR Test, version 1.5, were compared. CAP/CA reduced and consolidated labor while modestly increasing assay throughput without increased failure rates or direct costs, regardless of batch size and assay format.
Molecular Study of Microsporidiosis Due toEnterocytozoon bieneusiandEncephalitozoon intestinalisamong Human Immunodeficiency Virus-Infected Patients from Two Geographical Areas: Niamey, Niger, and Hanoi, Vietnam
Authors: Espern Anne , Morio Florent , Miegeville Michel , Illa Hachimou , Abdoulaye Moustapha , Meyssonnier Vanina , Adehossi Eric , Lejeune Anne , Cam Phung Dac , Besse Bernard , Gay-Andrieu Françoise 
Published: July 2007

ABSTRACTMicrosporidiosis cases due toEnterocytozoon bieneusiandEncephalitozoon intestinalisare emerging opportunistic infections associated with a wide range of clinical syndromes in humans. The aim of this study was to specify microsporidial epidemiology in two different geographical areas. From November 2004 to August 2005, 228 and 42 stool samples were collected in Niamey, Niger, and Hanoi, Vietnam, respectively. Screening for microsporidia was performed using UV-light microscopy. Detection was confirmed by molecular biology using two methods specific forE. bieneusiandE. intestinalis. All samples positive forE. bieneusiwere subjected to genotyping. In this study, we found high prevalences of microsporidiosis among human immunodeficiency virus-infected patients, 10.5% and 9.5%, respectively, in Niamey and Hanoi. These levels of prevalence are similar to those recorded in European countries before highly active antiretroviral therapy was introduced. In the samples positive forE. bieneusi, we found seven distinct genotypes, including two genotypes not previously described. TheE. bieneusigenotype distributions in the two geographical areas suggest different routes of infection transmission, person-to-person in Niger and zoonotic in Vietnam.
Molecular Epidemiological and Antibiotic Susceptibility Characterization of
Brucella
Isolates from Humans in Sicily, Italy
Authors: Marianelli Cinzia , Graziani Caterina , Santangelo Carmela , Xibilia Maria Teresa , Imbriani Alida , Amato Rosa , Neri Domenico , Cuccia Mario , Rinnone Sebastiano , Di Marco Vincenzo , Ciuchini Franco 
Published: July 2007

ABSTRACT

Brucellosis is a serious problem in Sicily.
Brucella melitensis
was identified as the species most frequently isolated in humans in Italy. No data, however, are available about the molecular epidemiological characterization of
Brucella
isolates from humans. We have conducted this study to molecularly characterize clinical isolates of
Brucella
spp. and to evaluate their antimicrobial susceptibilities. Twenty
Brucella
isolates were studied. Differential growth characteristics and DNA polymorphisms such as the restriction patterns of the PCR-amplified
omp2a
and
omp2b
genes,
rpoB
nucleotide sequencing, and multiple-locus variable-number tandem repeat analysis of 16 loci (MLVA-16) were used to characterize the strains. In vitro antibiotic susceptibility was determined by the E-test method on two different agar media, and the results were compared. All isolates were identified as
B. melitensis
biovar 3.
rpoB
nucleotide sequence analysis allowed the identification of two different genotypes of
B. melitensis
biovar 3. On the other hand, the MLVA-16 typing assay recognized 17 distinct genotypes. All isolates were sensitive to all tested antibiotics (rifampin, doxycycline, ciprofloxacin, ceftriaxone, and trimethoprim-sulfamethoxazole), and the Mueller-Hinton agar plate is recommended for antibiotic susceptibility testing by the E-test method. Our findings identify
B. melitensis
biovar 3 as the etiological agent isolated in Sicily and encourage the use of both molecular methods, and in particular of the MLVA-16 assay, in epidemiological trace-back analysis. This study represents the first epidemiological data from molecular typing of
Brucella
strains circulating in Italy and, in particular, in eastern Sicily.
Bacterial Peritonitis Caused byKingella kingae
Authors: Bofinger Jason J. , Fekete Thomas , Samuel Rafik 
Published: July 2007

ABSTRACTKingella kingaeis a commensal of the upper respiratory tract that occasionally causes skeletal infections in children and endocarditis in children and adults. We report a case of a 55-year-old man with liver disease and tense ascites who performed a paracentesis on himself and developedK. kingaeperitonitis and bacteremia.
The ORF49 Protein of Murine Gammaherpesvirus 68 Cooperates with RTA in Regulating Virus Replication
Authors: Lee Sangmi , Cho Hye-Jeong , Park Jung-Jin , Kim Yong-Sun , Hwang Seungmin , Sun Ren , Song Moon Jung 
Published: July 2007

ABSTRACT

Our functional mapping study of murine gammaherpesvirus 68 (MHV-68, or γHV-68) revealed that a mutant harboring a transposon at the ORF49 locus (ORF49
null
) evidenced a highly attenuated in vitro growth. ORF49 resides adjacent to and in an opposite direction from RTA, the primary switch of the gammaherpesvirus life cycle. A FLAG-tagged ORF49 protein was able to transcomplement ORF49
null
, and a revertant of ORF49
null
restored its attenuated growth to a level comparable to that of the wild type. The FLAG-tagged ORF49 protein promoted the ability of RTA to activate downstream target promoters and enhanced virus replication from the ORF50
null
virus in the presence of RTA. Furthermore, ORF49 enhanced wild-type virus replication by increasing the RTA transcript levels. Our data indicate that ORF49 may perform an important function in MHV-68 replication in cooperation with RTA.
A Tyrosine-Based Signal Plays a Critical Role in the Targeting and Function of Adenovirus RIDα Protein
Authors: Cianciola Nicholas L. , Crooks Denise , Shah Ankur H. , Carlin Cathleen 
Published: July 2007

ABSTRACT

Early region 3 genes of human adenoviruses contribute to the virus life cycle by altering the trafficking of cellular proteins involved in adaptive immunity and inflammatory responses. The ability of early region 3 genes to target specific molecules suggests that they could be used to curtail pathological processes associated with these molecules and treat human disease. However, this approach requires genetic dissection of the multiple functions attributed to early region 3 genes. The purpose of this study was to determine the role of targeting on the ability of the early region 3-encoded protein RIDα to downregulate the EGF receptor. A fusion protein between the RIDα cytoplasmic tail and glutathione
S
-transferase was used to isolate clathrin-associated adaptor 1 and adaptor 2 protein complexes from mammalian cells. Deletion and site-directed mutagenesis studies showed that residues 71-AYLRH of RIDα are necessary for in vitro binding to both adaptor complexes and that Tyr72 has an important role in these interactions. In addition, RIDα containing a Y72A point mutation accumulates in the
trans
-Golgi network and fails to downregulate the EGF receptor when it is introduced into mammalian cells as a transgene. Altogether, our data suggest a model where RIDα is trafficked directly from the
trans
-Golgi network to an endosomal compartment, where it intercepts EGF receptors undergoing constitutive recycling to the plasma membrane and redirects them to lysosomes.
Modification of Small RNAs Associated with Suppression of RNA Silencing by Tobamovirus Replicase Protein
Authors: Vogler Hannes , Akbergenov Rashid , Shivaprasad Padubidri V. , Dang Vy , Fasler Monika , Kwon Myoung-Ok , Zhanybekova Saule , Hohn Thomas , Heinlein Manfred 
Published: July 2007

ABSTRACT

Plant viruses act as triggers and targets of RNA silencing and have evolved proteins to suppress this plant defense response during infection. Although
Tobacco mosaic tobamovirus
(TMV) triggers the production of virus-specific small interfering RNAs (siRNAs), this does not lead to efficient silencing of TMV nor is a TMV-green fluorescent protein (GFP) hybrid able to induce silencing of a GFP-transgene in
Nicotiana benthamiana
, indicating that a TMV silencing suppressor is active and acts downstream of siRNA production. On the other hand, TMV-GFP is unable to spread into cells in which GFP silencing is established, suggesting that the viral silencing suppressor cannot revert silencing that is already established. Although previous evidence indicates that the tobamovirus silencing suppressing activity resides in the viral 126-kDa small replicase subunit, the mechanism of silencing suppression by this virus family is not known. Here, we connect the silencing suppressing activity of this protein with our previous finding that
Oilseed rape mosaic tobamovirus
infection leads to interference with HEN1-mediated methylation of siRNA and micro-RNA (miRNA). We demonstrate that TMV infection similarly leads to interference with HEN1-mediated methylation of small RNAs and that this interference and the formation of virus-induced disease symptoms are linked to the silencing suppressor activity of the 126-kDa protein. Moreover, we show that also
Turnip crinkle virus
interferes with the methylation of siRNA but, in contrast to tobamoviruses, not with the methylation of miRNA.
Epstein-Barr Virus Latent Nuclear Antigens Can Induce Metastasis in a Nude Mouse Model
Authors: Kaul Rajeev , Murakami Masanao , Choudhuri Tathagata , Robertson Erle S. 
Published: July 2007

ABSTRACT
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus associated with the development of both lymphoid and epithelial tumors. The EBV critical latent antigens EBNA1 and EBNA3C interact with Nm23-H1, a known suppressor of cell migration and tumor metastasis. This interaction is critical for the regulation of downstream cellular genes involved in tumorigenesis and cell migration. The significance of these interactions was determined in nude mice using cancer cells expressing both EBV antigens and Nm23-H1. The EBV antigens promoted the growth of transformed cells in vivo, but their expression was less critical during the later stage of tumor development. The expression of Nm23-H1 affected the growth of cancer cells and suppressed their metastatic potential. This effect was effectively rescued by the expression of both EBV antigens. Interestingly, the prometastatic potential of EBNA3C was greater than that of EBNA1, which triggered a dramatic immune response, as indicated by increased spleen size and development of ascites in the mice. These studies now bridge the expression of the EBV antigens with tumorigenesis and metastasis and widen the range of potential targets for development of therapies for EBV-associated malignancies.
Murine Gammaherpesvirus 68
ORF52
Encodes a Tegument Protein Required for Virion Morphogenesis in the Cytoplasm
Authors: Bortz Eric , Wang Lili , Jia Qingmei , Wu Ting-Ting , Whitelegge Julian P. , Deng Hongyu , Zhou Z. Hong , Sun Ren 
Published: July 2007

ABSTRACT

The tegument, a semiordered matrix of proteins overlying the nucleocapsid and underlying the virion envelope, in viruses in the gamma subfamily of
Herpesviridae
is poorly understood. Murine gammaherpesvirus 68 (MHV-68) is a robust model for studying gammaherpesvirus virion structure, assembly, and composition, as MHV-68 efficiently completes the lytic phase and productively infects cultured cells. We have found that MHV-68
ORF52
encodes an abundant tegument protein conserved among gammaherpesviruses. Detergent sensitivity experiments revealed that the MHV-68 ORF52 protein is more tightly bound to the virion nucleocapsid than the ORF45 tegument protein but could be dissociated from particles that retained the ORF65 small capsomer protein. ORF52, tagged with enhanced green fluorescent protein or FLAG epitope, localized to the cytoplasm. A recombinant MHV-68 bacterial artificial chromosome mutant with a nonsense mutation incorporated into
ORF52
exhibited viral DNA replication, expression of late lytic genes, and capsid assembly and packaging at levels near those of the wild type. However, the MHV-68
ORF52
-null virus was deficient in the assembly and release of infectious virion particles. Instead, partially tegumented capsids produced by the
ORF52
-null mutant accumulated in the cytoplasm, containing conserved capsid proteins, the ORF64 and ORF67 tegument proteins, but virtually no ORF45 tegument protein. Thus, ORF52 is essential for the tegumentation and egress of infectious MHV-68 particles in the cytoplasm, suggesting an important conserved function in gammaherpesvirus virion morphogenesis.