bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Arabidopsis Calcium Sensor Calcineurin B-Like 3 Inhibits the 5′-Methylthioadenosine Nucleosidase in a Calcium-Dependent Manner
Authors: Oh Seung-Ick , Park Jimyeong , Yoon Sunhee , Kim Yungyeong , Park Soojin , Ryu Migyeong , Nam Min Jung , Ok Sung Han , Kim Jeong-Kook , Shin Jeong-Sheop , Kim Kyung-Nam 
Published: October 2008

Abstract
Calcineurin B-like (CBL) proteins represent a unique family of calcium sensors in plant cells. Sensing the calcium signals elicited by a variety of abiotic stresses, CBLs transmit the information to a group of serine/threonine protein kinases (CBL-interacting protein kinases [CIPKs]), which are currently known as the sole targets of the CBL family. Here, we report that the CBL3 member of this family has a novel interaction partner in addition to the CIPK proteins. Extensive yeast two-hybrid screenings with CBL3 as bait identified an interesting Arabidopsis (Arabidopsis thaliana) cDNA clone (named AtMTAN, for 5′-methylthioadenosine nucleosidase), which encodes a polypeptide similar to EcMTAN from Escherichia coli. Deletion analyses showed that CBL3 utilizes the different structural modules to interact with its distinct target proteins, CIPKs and AtMTAN. In vitro and in vivo analyses verified that CBL3 and AtMTAN physically associate only in the presence of Ca2+. In addition, we empirically demonstrated that the AtMTAN protein indeed possesses the MTAN activity, which can be inhibited specifically by Ca2+-bound CBL3. Overall, these findings suggest that the CBL family members can relay the calcium signals in more diverse ways than previously thought. We also discuss a possible mechanism by which the CBL3-mediated calcium signaling regulates the biosynthesis of ethylene and polyamines, which are involved in plant growth and development as well as various stress responses.
Development of a Nucleocapsid-Based Human Coronavirus Immunoassay and Estimates of Individuals Exposed to Coronavirus in a U.S. Metropolitan Population
Authors: Severance Emily G. , Bossis Ioannis , Dickerson Faith B. , Stallings Cassie R. , Origoni Andrea E. , Sullens Anne , Yolken Robert H. , Viscidi Raphael P. 
Published: October 2008

ABSTRACTCoronaviruses cause respiratory infections ranging from common colds to severe acute respiratory syndrome (SARS) in humans. Estimates for exposure to non-SARS coronaviruses are high, particularly for 229E and OC43; however, less information regarding seroprevalence is available for HKU1 and NL63. To measure exposure rates to these four coronavirus strains (229E, HKU1, NL63, and OC43), we devised an immunoassay based on amino- and carboxy-terminally tagged recombinant coronavirus nucleocapsid antigens. Four human and one feline coronavirus antigen were cloned into baculoviruses expressed in insect cells and recovered proteins bound in the solid phase of an enzyme-linked immunosorbent assay-based system. We screened sera from 10 children and 196 adults and established primary cutoff points based on immunoglobulin G (IgG) antibody levels of the predominantly seronegative children. The proportion of seropositive adults for each coronavirus was as follows: 229E, 91.3%; HKU1, 59.2%; NL63, 91.8%; and OC43, 90.8%. No evidence of a significant serological response to the feline coronavirus was observed. Significant associations of coronavirus seropositivity and antibody levels with age, gender, race, socioeconomic status, smoking status, and season of the blood draw were tested with chi-square and regression analyses. The group II coronaviruses (OC43 and HKU1) were significantly associated with race (P≤ 0.009 andP≤ 0.03, respectively). Elevated OC43 IgG levels were further significantly associated with smoking status (P≤ 0.03), as were high NL63 titers with socioeconomic status (P≤ 0.04). The high-level immunoreactivity of each coronavirus was significantly associated with the summer season (P≤ 0.01 to 0.0001). In summary, high rates of exposure to 229E, NL63, and OC43 and a moderate rate of exposure to HKU1 characterized the seroprevalence among individuals in this population. Demographic factors, such as race, smoking status, and socioeconomic status, may confer an increased risk of susceptibility to these viruses.
Serological Diagnosis of Leprosy in Patients in Vietnam by Enzyme-Linked Immunosorbent Assay with
Mycobacterium leprae
-Derived Major Membrane Protein II
Authors: Kai Masanori , Nguyen Phuc Nhu Ha , Hoang Thi Thuy Huong , Nguyen An Hoang , Fukutomi Yasuo , Maeda Yumi , Miyamoto Yuji , Mukai Tetsu , Fujiwara Tsuyoshi , Nguyen Tan Thanh , Makino Masahiko 
Published: October 2008

ABSTRACT
A serological diagnostic test using phenolic glycolipid-I (PGL-I) developed in the 1980s is commercially available, but the method is still inefficient in detecting all forms of leprosy. Therefore, more-specific and -reliable serological methods have been sought. We have characterized major membrane protein II (MMP-II) as a candidate protein for a new serological antigen. In this study, we evaluated the effectiveness of the enzyme-linked immunosorbent assay (ELISA) using the MMP-II antigen (MMP-II ELISA) for detecting antibodies in leprosy patients and patients' contacts in the mid-region of Vietnam and compared to the results to those for the PGL-I method (PGL-I ELISA). The results showed that 85% of multibacillary patients and 48% of paucibacillary patients were positive by MMP-II ELISA. Comparison between the serological tests showed that positivity rates for leprosy patients were higher with MMP-II ELISA than with PGL-I ELISA. Household contacts (HHCs) showed low positivity rates, but medical staff members showed comparatively high positivity rates, with MMP-II ELISA. Furthermore, monitoring of results for leprosy patients and HHCs showed that MMP-II is a better index marker than PGL-I. Overall, the epidemiological study conducted in Vietnam suggests that serological testing with MMP-II would be beneficial in detecting leprosy.
Downregulation of CD40 Ligand Response in Monocytes from Sepsis Patients
Authors: Sinistro Anna , Almerighi Cristiana , Ciaprini Chiara , Natoli Silvia , Sussarello Emanuele , Di Fino Sara , Calò-Carducci Francesca , Rocchi Giovanni , Bergamini Alberto 
Published: October 2008

ABSTRACTIt has been suggested that a defective adaptive immune response contributes to septic immunosuppression. Here, the response of monocytes to CD40 ligand (CD40L) for patients with sepsis due to infection with gram-negative organisms has been analyzed. Compared to cells from controls, monocytes from septic patients showed significantly reduced production of tumor necrosis factor alpha, interleukin-1β (IL-1β), and IL-12 and were unable to acquire high levels of CD80 and CD86 molecules. These alterations were observed at the onset of sepsis and persisted at day 7. However, the ability of monocytes to respond to CD40L stimulation was partially but significantly restored in cells from patients who recovered from sepsis. In addition, costimulation of autologous CD4+T lymphocytes by CD40L-activated monocytes from septic patients failed to induce cell proliferation and gamma interferon production. Finally, the ability of CD40L to rescue monocytes from apoptosis was severely impaired. We conclude that downregulation of the CD40L response may be an appropriate model for the monocyte alteration observed during septic immunosuppression and may help in the development of novel therapeutic strategies.
Evaluation of the Recombinant VlsE-Based Liaison Chemiluminescence Immunoassay for Detection ofBorrelia burgdorferiand Diagnosis of Lyme Disease
Authors: Ledue Thomas B. , Collins Marilyn F. , Young John , Schriefer Martin E. 
Published: October 2008

ABSTRACTRecent efforts to improve the serologic diagnosis of Lyme disease have included the use of a synthetic peptide (C6) that reproduces the sequence of invariable region 6 of VlsE, the variable surface antigen ofBorrelia burgdorferi. In the present study, the diagnostic performance of DiaSorin's recombinant VlsE-based chemiluminescence immunoassay in 1,947 human serum samples was evaluated. Sensitivity was determined using two serum panels from the CDC. For panel I, we observed sensitivities of 68.4% and 75.6% for subjects with early, localized (n= 19) or disseminated (n= 41) disease, respectively. For panel II, we observed sensitivities of 61.5% and 100% for subjects with early (n= 26) or late-stage (n= 11) disease, respectively. We observed a specificity of 99.5% for healthy donors (n= 600) living either in regions of the United States where the disease is endemic or in regions where it is not endemic. Overall, specificity among 207 potentially cross-reactive sera from subjects who had other spirochetal infections, nonspirochetal infections including bacterial and viral infections, or autoimmune or neurologic disease; who were positive for rheumatoid factor or anti-mouse antibodies; or who had been previously vaccinated for Lyme disease was 93.7%. In a direct comparison of 1,038 prospectively collected samples for Lyme disease testing we observed a relative sensitivity of 70%, a relative specificity of 99.1%, and an overall agreement of 97.1% between the DiaSorin recombinant VlsE chemiluminescence immunoassay and the Immunetics peptide-based C6 enzyme-linked immunosorbent assay.
Improved Sensitivity of Diagnosis of Tuberculosis in Patients in Korea via a Cocktail Enzyme-Linked Immunosorbent Assay Containing the Abundantly Expressed Antigens of the K Strain ofMycobacterium tuberculosis
Authors: Shin A-Rum , Shin Sung Jae , Lee Kil-Soo , Eom Sun-Ho , Lee Seung-Sub , Lee Byung-Soo , Lee Ji-Sook , Cho Sang Nae , Kim Hwa-Jung 
Published: October 2008

ABSTRACTTuberculosis (TB) is the leading cause of death from a single infectious agent in Korea. In this study, we compared the proteins present in culture filtrates fromMycobacterium tuberculosisstrain K, which is the dominant clinical isolate in Korea, with those present in culture filtrates fromM. tuberculosisH37Rv. Several differences in expression were detected between the two strains for those proteins with a molecular mass of <20 kDa. ESAT-6, HSP-X, and CFP-10 were found to be abundantly expressed in the strain K culture filtrates by liquid chromatography-electrospray ionization-time of flight mass spectrometry. The serodiagnostic potentials of recombinant antigens rESAT-6, rHSP-X, and rCFP-10 and two native antigens (Ag85 and PstS1) were evaluated by Western blot analysis and enzyme-linked immunosorbent assay (ELISA) using sera collected from 46 TB patients with active disease and 46 healthy controls. As for our ELISA results, HSP-X was superior to the other antigens in terms of sensitivity when a single antigen was employed. The results of a receiver operator characteristic analysis revealed that a cocktail ELISA using all five antigens was significantly more sensitive (77.8%) than the use of a single antigen and offered equivalent specificity; moreover, it produced the largest area under the curve (0.91 versus 0.55 to 0.87). Therefore, a cocktail ELISA containing abundantly expressed antigens enhances the sensitivity of a single antigen and can be a useful diagnostic tool for the detection of active TB.
Real-Time In Vivo Green Fluorescent Protein Imaging of a Murine Leishmaniasis Model as a New Tool for
Leishmania
Vaccine and Drug Discovery
Authors: Mehta Sanjay R. , Huang Robert , Yang Meng , Zhang Xing-Quan , Kolli Bala , Chang Kwang-Poo , Hoffman Robert M. , Goto Yasuyuki , Badaro Roberto , Schooley Robert T. 
Published: October 2008

ABSTRACT

Leishmania
species are obligate intracellular protozoan parasites that cause a broad spectrum of clinical diseases in mammalian hosts. The most frequently used approach to quantify parasites in murine model systems is based on thickness measurements of the footpad or ear after experimental infection. To overcome the limitations of this method, we used a
Leishmania
mutant episomally transfected with enhanced green fluorescent protein, enabling in vivo real-time whole-body fluorescence imaging, to follow the progression of
Leishmania
infection in parasitized tissues. Fluorescence correlated with the number of
Leishmania
parasites in the tissue and demonstrated the real-time efficacy of a therapeutic vaccine. This approach provides several substantial advantages over currently available animal model systems for the in vivo study of immunopathogenesis, prevention, and therapy of leishmaniasis. These include improvements in sensitivity and the ability to acquire real-time data on progression and spread of the infection.
Evaluation of Antigen-Specific T-Cell Responses with a Miniaturized and Automated Method
Authors: Li Pira Giuseppina , Ivaldi Federico , Dentone Chiara , Righi Elda , Del Bono Valerio , Viscoli Claudio , Koopman Gerrit , Manca Fabrizio 
Published: October 2008

ABSTRACTThe evaluation of antigen-specific T-cell responses is helpful for both research and clinical settings. Several techniques can enumerate antigen-responsive T cells or measure their products, but they require remarkable amounts of peripheral blood mononuclear cells (PBMCs). Since screening numerous antigens or testing samples from pediatric or lymphopenic patients is hampered in clinical practice, we refined a miniaturized, high-throughput assay for T-cell immunity. Antigens and cells in 10-μl volumes were dispensed into 1,536-well culture plates precoated with anti-gamma interferon (anti-IFN-γ) antibodies. After being cultured, the wells were developed by enzyme-linked immunosorbent assay for bound cytokine. Miniaturization and automation allowed quantitation of antigen-specific responses on 104PBMCs. This method was applied for epitope mapping of mycobacterial antigens and was used in the clinic to evaluate T-cell immunity to relevant opportunistic pathogens by using small blood samples. A comparison with conventional methods showed similar sensitivity. Therefore, current flow cytometric methods that provide information on frequency and phenotype of specific T cells can be complemented by this assay that provides extensive information on cytokine concentrations and profiles and requires 20- to 50-fold fewer PBMCs than other analytical methods.
Characterization of Extensively Drug-Resistant
Mycobacterium tuberculosis
Clinical Isolates in China
Authors: Sun Zhaogang , Chao Yanjie , Zhang Xuxia , Zhang Jianyuan , Li Yan , Qiu Yunqing , Liu Yi , Nie Lihui , Guo Aizhen , Li Chuanyou 
Published: October 2008

ABSTRACT
Thirteen extensively drug-resistant tuberculosis isolates which were highly resistant to a broad spectrum of antituberculosis drugs were identified from 1,926 clinical isolates in China. They had highly diverse mycobacterial interspersed repetitive-unit-variable-number tandem-repeat patterns. Most, but not all, of the drug target genes had mutations contributing to resistance to the corresponding drug.
Molecular Epidemiology and Phylogenetic Distribution of the
Escherichia coli pks
Genomic Island
Authors: Johnson James R. , Johnston Brian , Kuskowski Michael A. , Nougayrede Jean-Philippe , Oswald Eric 
Published: October 2008

ABSTRACT

Epidemiological and phylogenetic associations of the
pks
genomic island of extraintestinal pathogenic
Escherichia coli
(ExPEC), which encodes the genotoxin colibactin, are incompletely defined.
clbB
and
clbN
(as markers for the 5′ and 3′ regions of the
pks
island, respectively),
clbA
and
clbQ
(as supplemental
pks
island markers), and 12 other putative ExPEC virulence genes were newly sought by PCR among 131 published
E. coli
isolates from hospitalized veterans (62 blood isolates and 69 fecal isolates). Blood and fecal isolates and
clbB
-positive and -negative isolates were compared for 66 newly and previously assessed traits. Among the 14 newly sought traits,
clbB
and
clbN
(colibactin polyketide synthesis system),
hra
(heat-resistant agglutinin), and
vat
(vacuolating toxin) were significantly associated with bacteremia.
clbB
and
clbN
identified a subset within phylogenetic group B2 with extremely high virulence scores and a high proportion of blood isolates. However, by multivariable analysis, other traits were more predictive of blood source than
clbB
and
clbN
were; indeed, among the newly sought traits, only
pic
significantly predicted bacteremia (negative association). By correspondence analysis,
clbB
and
clbN
were closely associated with group B2 and multiple B2-associated traits; by principal coordinate analysis,
clbB
and
clbN
partitioned the data set better than did blood versus fecal source. Thus, the
pks
island was significantly associated with bacteremia, multiple ExPEC-associated virulence genes, and group B2, and within group B2, it identified an especially high-virulence subset. This extends previous work regarding the
pks
island and supports investigation of the colibactin system as a potential therapeutic target.
Multiple-Host Sharing, Long-Term Persistence, and Virulence of
Escherichia coli
Clones from Human and Animal Household Members
Authors: Johnson James R. , Clabots Connie , Kuskowski Michael A. 
Published: October 2008

ABSTRACT

During a 3-year surveillance, six household members (five humans and the family dog) yielded 14
Escherichia coli
clones. Virulence genes, group B2, and having caused cystitis (in the mother or dog) corresponded to colonization endpoints (number of samples, colonies, hosts, and dates). The dog's cystitis clone was the most extensively recovered clone.
Development and Evaluation of a Molecular Assay for Detection of Nontuberculous Mycobacteria by Use of the Cobas Amplicor Platform
Authors: Peter-Getzlaff S. , Lüthy J. , Böddinghaus B. , Böttger E. C. , Springer B. 
Published: October 2008

ABSTRACT

We have developed and evaluated a semiautomated assay for detection of nontuberculous mycobacteria (NTM) from clinical samples based on the Cobas Amplicor
Mycobacterium tuberculosis
test (Roche Diagnostics, Switzerland). A capture probe, specific for mycobacteria at the genus level, was linked to magnetic beads and used for the detection of amplification products obtained by the Cobas Amplicor
M. tuberculosis
assay. We demonstrate that the analytical sensitivity of the genus assay is similar to that of Cobas Amplicor
M. tuberculosis
detection. Four hundred sixteen clinical specimens were evaluated for the presence of NTM DNA. Sensitivities for smear-positive and smear-negative specimens were found to be 100% and 47.9%, respectively. Specificity was 97.7%, the positive predictive value 84.6%, and the negative predictive value 93.1%. The genus assay is easy to perform, produces reliable results, and was found to be a valuable diagnostic tool for rapid diagnosis of infections with NTM. The genus assay has the potential to detect NTM not routinely recovered by culture and to discover new mycobacterial species.
Comparative Study for Determination of
Mycobacterium tuberculosis
Susceptibility to First- and Second-Line Antituberculosis Drugs by the Etest Using 7H11, Blood, and Chocolate Agar
Authors: Coban Ahmet Yilmaz , Bilgin Kemal , Uzun Meltem , Akgunes Alper , Yusof Anne , Durupinar Belma 
Published: October 2008

ABSTRACT

We investigated the performance of blood and chocolate agar as alternatives to Middlebrook 7H11 agar for testing the susceptibility of
Mycobacterium tuberculosis
to first-and second-line drugs by the Etest method. A total of 39 strains of
M. tuberculosis
including 22 multidrug-resistant
M. tuberculosis
strains and 17 susceptible strains were tested. In conclusion, our results showed that chocolate agar gave insufficient growth, needing up to 21 days of incubation, while results on blood agar were comparable to those on Middlebrook 7H11 agar and can be further explored as an alternative for Etest-based susceptibility testing of
M. tuberculosis
.
Rapid Detection of
Mycobacterium tuberculosis
Resistance to Second-Line Drugs by Use of the Manual Mycobacterium Growth Indicator Tube System
Authors: Martin Anandi , von Groll Andrea , Fissette Krista , Palomino Juan Carlos , Varaine Francis , Portaels Françoise 
Published: October 2008

ABSTRACT

The objective of this study was to evaluate the manual mycobacterium growth indicator tube (MGIT) system for the testing of
Mycobacterium tuberculosis
susceptibility to second-line drugs compared to the proportion method. One hundred eighty-eight
M. tuberculosis
isolates were tested for susceptibility to ofloxacin, kanamycin, ethionamide, and capreomycin by the manual MGIT, and results were compared to those obtained with the proportion method on 7H11 agar, considered a reference method. Results for ofloxacin and capreomycin were excellent, with 100% accuracy, and a result of 99.4% accuracy was achieved for kanamycin. For ethionamide, accuracy was lower, with a result of 86.7% compared to that of the proportion method. We proposed the following critical concentrations for the drugs: for ofloxacin, 2.0 μg/ml; for kanamycin, 2.5 μg/ml; for ethionamide, 5 μg/ml; and for capreomycin, 2.5 μg/ml. The time required to obtain results was an average of 8 days by the manual MGIT and 3 weeks by the reference method. Our results show that the manual MGIT is an accurate method for the rapid susceptibility testing of
M. tuberculosis
to second-line drugs. There is no need for a machine when using the manual MGIT, and results can be read with a simple UV lamp or with a semiquantitative reader, which considerably reduces the cost of the method.
Genotypic Antiretroviral Resistance Testing for Human Immunodeficiency Virus Type 1 Integrase Inhibitors by Use of the TruGene Sequencing System
Authors: Paar Christian , Palmetshofer Christina , Flieger Karin , Geit Maria , Kaiser Rolf , Stekel Herbert , Berg Jörg 
Published: October 2008

ABSTRACTA sequencing assay for detection of mutations conferring resistance to human immunodeficiency virus type 1 (HIV-1) integrase inhibitors raltegravir and elvitegravir was developed using the automated TruGene sequencing system. The assay returned clear sequencing results for samples with ≥500 RNA copies/ml for mutation detection and HIV-1 subtyping across a spectrum of HIV-1 subtypes.
High Prevalence of Human Parechovirus (HPeV) Genotypes in the Amsterdam Region and Identification of Specific HPeV Variants by Direct Genotyping of Stool Samples
Authors: Benschop K. , Thomas X. , Serpenti C. , Molenkamp R. , Wolthers K. 
Published: October 2008

ABSTRACT

Human parechoviruses (HPeV) are widespread pathogens belonging to the
Picornavirus
family. Six genotypes, which have predominantly been isolated from children, are known. Data on prevalence of HPeV genotypes are generally based on cell culture, which may underestimate the prevalence of certain HPeV strains that are difficult to grow. We studied 1,824 stool samples from 1,379 children (<5 years old) sent to the Academic Medical Center, Amsterdam, The Netherlands, between 2004 and 2006. Samples were screened using specific human enterovirus (HEV) and HPeV real-time PCRs based on the 5′ untranslated region. A high percentage of HPeV infections (16.3%), comparable to the percentage of HEV infections (18.4%), were found by PCR in stool samples. HPeV-positive stool samples were directly genotyped based on the VP1 region for the first time to avoid a culture bias. HPeV1 was found to be the most prevalent type. The majority of the HPeV1 strains clustered separately from the prototype strain, Harris, which has not been reported to circulate lately. However, we could identify three strains as HPeV1 Harris. HPeV3 was identified as the second most predominant type during 2004 and 2006 but was not found in 2005. HPeV4 to -6 were found in smaller numbers. One strain could not be associated with a known HPeV type (VP1 gene nucleotide similarity: 71%), possibly indicating a new genotype. Also, we report the first identification of three HPeV5 strains and one HPeV1 strain with a different motif at the C-terminal end of VP1, where the arginine-glycine-aspartic acid (RGD) motif is normally located.