bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunization with Oligomannose-Coated Liposome-Entrapped Dense Granule Protein 7 Protects Dams and Offspring fromNeospora caninumInfection in Mice
Authors: Nishikawa Yoshifumi , Zhang Houshuang , Ikehara Yuzuru , Kojima Naoya , Xuan Xuenan , Yokoyama Naoaki 
Published: April 2009

ABSTRACTThe present study demonstrates that the subcutaneous administration ofNeospora caninumdense granule protein 7 (NcGRA7) entrapped in liposomes coated with mannotriose strongly induces the parasite-specific T-helper type 1 immune response and humoral antibody in mice. Although anti-NcGRA7 immunoglobulin G1 antibody production was induced in mice injected with NcGRA7 alone, the dams and offspring were never protected fromN. caninuminfection. The immunization of mice with liposome-entrapped NcGRA7 before pregnancy resulted in increased offspring survival and decreased the infection rates in the brains of dams after parasite infection at 6 to 9 days of gestation. In conclusion, oligomannose-coated liposome-entrapped NcGRA7 can be used as a new type of effective vaccine to control neosporosis.
Real-Time Quantitative PCR To Determine Chlamydial Load in Men and Women in a Community Setting
Authors: Wiggins R. , Graf S. , Low N. , Horner P. J. 
Published: April 2009

ABSTRACT

We used a PCR method to quantify the loads of
Chlamydia trachomatis
organisms in self-collected urine and vulvovaginal swab (VVS) samples from 93 women and 30 men participating in the Chlamydia Screening Studies Project, a community-based study of individuals not seeking health care. For women, self-collected VVS had a higher mean chlamydial load (10,405 organisms/ml; 95% confidence interval [95% CI], 5,167 to 21,163 organisms/ml) than did first-void urines (FVU) (503 organisms/ml; 95% CI, 250 to 1,022 organisms/ml;
P
< 0.001). Chlamydial loads in female and male self-collected FVU specimens were similar (
P
= 0.634). The mean chlamydial load in FVU specimens decreased with increasing age in females and males. There was no strong statistical evidence of differences in chlamydial load in repeat male and female FVU specimens taken when patients attended for treatment a median of 23.5 (range, 14 to 62) and 28 (range, 13 to 132) days later, respectively, or in VVS taken a median of 35 (range, 14 to 217) days later. In this study, chlamydial load values for infected persons in the community who were not seeking treatment were lower than those published in other studies involving symptomatic patients attending clinical settings. This might have implications for estimates of the infectiousness of chlamydia. The results of this study provide a scientific rationale for preferring VVS to FVU specimens from women.
Simultaneous Identification of 14 Genital Microorganisms in Urine by Use of a Multiplex PCR-Based Reverse Line Blot Assay
Authors: Mckechnie Michelle L. , Hillman Richard , Couldwell Deborah , Kong Fanrong , Freedman Eleanor , Wang Hui , Gilbert Gwendolyn L. 
Published: April 2009

ABSTRACT

The aim of this study was to develop and evaluate a sensitive method for the simultaneous identification of 14 urogenital potential pathogens. A multiplex PCR-based reverse line blot (mPCR/RLB) assay was developed to detect 14 urogenital pathogens or putative pathogens, namely
Trichomonas vaginalis
,
Streptococcus pneumoniae
,
Neisseria gonorrhoeae
,
Chlamydia trachomatis
,
Ureaplasma parvum
,
U. urealyticum
,
Gardnerella vaginalis
,
Haemophilus influenzae
, herpes simplex virus type 1 (HSV1) and HSV2,
N. meningitidis
,
Mycoplasma hominis
,
M. genitalium
, and adenovirus, using two species-specific primer pairs and probes for each. The method was validated using a reference strain or a well-characterized clinical isolate of each target organism and was found to be both sensitive and specific. The limits of detection for the mPCR/RLB assay varied among the 14 target organisms from 4.2 × 10
−1
to 7.0 × 10
−11
ng/μl of genomic DNA. There were no cross-reactions among any of the probes. This method was used to test 529 first-voided urine specimens from male patients with and without urethritis attending two Sydney sexual health clinics. One or more target species were detected in 193 (36%) subjects. Of 233 positive results, overall 216 (93%) were concordant between mPCR/RLB and a comparator method (culture and/or species-specific PCR), 9 were positive only by mPCR/RLB, and 8 were positive only by the comparator method. The mPCR/RLB method was an accurate, convenient, and inexpensive method for the detection of multiple potential pathogens in first-voided urine specimens from men.
Distribution of the
Escherichia coli
Common Pilus among Diverse Strains of Human Enterotoxigenic
E. coli
Authors: Blackburn Dana , Husband Amanda , Saldaña Zeus , Nada Rania A. , Klena John , Qadri Firdausi , Girón Jorge A. 
Published: April 2009

ABSTRACT

The
Escherichia coli
common pilus (ECP) is produced by commensal and pathogenic
E. coli
strains. This pilus is unrelated to any of the known colonization factors (CFs) of enterotoxigenic
E. coli
(ETEC). In this study, we investigated the distribution and production of ECP among a collection of 136 human CF-positive and CF-negative ETEC strains of different geographic origins. The major pilus subunit gene,
ecpA
, was found in 109 (80%) of these strains, suggesting that it is widely distributed among ETEC strains. Phenotypic analysis of a subset of 43 strains chosen randomly showed that 58% of them produced ECP independently of the presence or absence of CFs, a percentage even higher than that of the most prevalent CFs. These data suggest an important role for ECP in the biology of ETEC, particularly in CF-negative strains, and in human infection.
Analysis of the Varicella-Zoster Virus IE62 N-Terminal Acidic Transactivating Domain and Its Interaction with the Human Mediator Complex
Authors: Yamamoto Shinobu , Eletsky Alexander , Szyperski Thomas , Hay John , Ruyechan William T. 
Published: April 2009

ABSTRACT
The varicella-zoster virus major transactivator, IE62, contains a potent N-terminal acidic transcriptional activation domain (TAD). Our experiments revealed that the minimal IE62 TAD encompasses amino acids (aa) 19 to 67. We showed that the minimal TAD interacts with the human Mediator complex. Site-specific mutations revealed residues throughout the minimal TAD that are important for both activation and Mediator interaction. The TAD interacts directly with aa 402 to 590 of the MED25 subunit, and site-specific TAD mutations abolished this interaction. Two-dimensional nuclear magnetic resonance spectroscopy revealed that the TAD is intrinsically unstructured. Our studies suggest that transactivation may involve the TAD adopting a defined structure upon binding MED25.
The Herpes Simplex Virus Type 1 Glycoprotein D (gD) Cytoplasmic Terminus and Full-Length gE Are Not Essential and Do Not Function in a Redundant Manner for Cytoplasmic Virion Envelopment and Egress
Authors: Lee Hyun Cheol , Chouljenko Vladimir N. , Chouljenko Dmitry V. , Boudreaux Marc J. , Kousoulas K. G. 
Published: April 2009

ABSTRACT

Herpes simplex virus type 1 (HSV-1) acquires its final envelope by budding into cytoplasmic vesicles thought to be derived from
trans
-Golgi network membranes. This process is facilitated by interactions among the carboxyl termini of viral glycoproteins and tegument proteins. To directly investigate the relative importance of the carboxyl terminus of glycoprotein D (gD) in the presence or absence of gE, a recombinant virus (gDΔct) was constructed to specify a truncated gD lacking the carboxy-terminal 29 amino acids. Furthermore, two additional recombinant viruses were constructed by mutating from ATG to CTG the initiation codons of gE (gEctg) or both gE and gM (gEctg+gMctg), causing lack of expression of gE or both gE and gM, respectively. A fourth mutant virus was constructed to specify the gEctg+gDΔct mutations. The replication properties of these viruses were compared to those of a newly constructed recombinant virus unable to express UL20 due to alteration of the two initiation codons of UL20 (UL20ctgctg). All recombinant viruses were constructed by using the double-Red, site-directed mutagenesis system implemented on the HSV-1(F) genome cloned into a bacterial artificial chromosome. The gEctg, gEctg+gMctg, gDΔct, and gEctg+gDΔct viruses produced viral plaques on African monkey kidney cells (Vero), as well as other cells, that were on average approximately 30 to 50% smaller than those produced by the wild-type virus HSV-1(F). In contrast, the UL20ctgctg virus produced very small plaques containing three to five cells, as reported previously for the ΔUL20 virus lacking the entire UL20 gene. Viral replication kinetics of intracellular and extracellular viruses revealed that all recombinant viruses produced viral titers similar to those produced by the wild-type HSV-1(F) virus intracellularly and extracellularly at late times postinfection, with the exception of the UL20ctgctg and ΔUL20 viruses, which replicated more than two-and-a-half logs less efficiently than HSV-1(F). Electron microscopy confirmed that all viruses, regardless of their different gene mutations, efficiently produced enveloped virions within infected cells, with the exception of the UL20ctgctg and ΔUL20 viruses, which accumulated high levels of unenveloped virions in the cytoplasm. These results show that the carboxyl terminus of gD and the full-length gE, either alone or in a redundant manner, are not essential in cytoplasmic virion envelopment and egress from infected cells. Similarly, gM and gE do not function alone or in a redundant manner in cytoplasmic envelopment and virion egress, confirming previous findings.
Bovine Coronavirus Nonstructural Protein 1 (p28) Is an RNA Binding Protein That Binds Terminal Genomic
cis
-Replication Elements
Authors: Gustin Kortney M. , Guan Bo-Jhih , Dziduszko Agnieszka , Brian David A. 
Published: April 2009

ABSTRACT

Nonstructural protein 1 (nsp1), a 28-kDa protein in the bovine coronavirus (BCoV) and closely related mouse hepatitis coronavirus, is the first protein cleaved from the open reading frame 1 (ORF 1) polyprotein product of genome translation. Recently, a 30-nucleotide (nt)
cis
-replication stem-loop VI (SLVI) has been mapped at nt 101 to 130 within a 288-nt 5′-terminal segment of the 738-nt nsp1 cistron in a BCoV defective interfering (DI) RNA. Since a similar nsp1 coding region appears in all characterized groups 1 and 2 coronavirus DI RNAs and must be translated in
cis
for BCoV DI RNA replication, we hypothesized that nsp1 might regulate ORF 1 expression by binding this intra-nsp1 cistronic element. Here, we (i) establish by mutation analysis that the 72-nt intracistronic SLV immediately upstream of SLVI is also a DI RNA
cis
-replication signal, (ii) show by gel shift and UV-cross-linking analyses that cellular proteins of ∼60 and 100 kDa, but not viral proteins, bind SLV and SLVI, (SLV-VI) and (iii) demonstrate by gel shift analysis that nsp1 purified from
Escherichia coli
does not bind SLV-VI but does bind three 5′ untranslated region (UTR)- and one 3′ UTR-located
cis
-replication SLs. Notably, nsp1 specifically binds SLIII and its flanking sequences in the 5′ UTR with ∼2.5 μM affinity. Additionally, under conditions enabling expression of nsp1 from DI RNA-encoded subgenomic mRNA, DI RNA levels were greatly reduced, but there was only a slight transient reduction in viral RNA levels. These results together indicate that nsp1 is an RNA-binding protein that may function to regulate viral genome translation or replication but not by binding SLV-VI within its own coding region.
Human Metapneumovirus Establishes Persistent Infection in the Lungs of Mice and Is Reactivated by Glucocorticoid Treatment
Authors: Liu Yuru , Haas Debra L. , Poore Spencer , Isakovic Sanjin , Gahan Michelle , Mahalingam Suresh , Fu Zhen F. , Tripp Ralph A. 
Published: April 2009

ABSTRACT
Human metapneumovirus (HMPV) has been identified as a worldwide agent of serious upper and lower respiratory tract infections in infants and young children. HMPV is second only to respiratory syncytial virus (RSV) as a leading cause of bronchiolitis, and, like RSV, consists of two major genotypes that cocirculate and vary among communities year to year. Children who have experienced acute HMPV infection may develop sequelae of wheezing and asthma; however, the features contributing to this pathology remain unknown. A possible mechanism for postbronchiolitis disease is that HMPV might persist in the lung providing a stimulus that could contribute to wheezing and asthma. Using immunohistochemistry to identify HMPV-infected cells in the lungs of mice, we show that HMPV mediates biphasic replication in respiratory epithelial cells then infection migrates to neuronal processes that innervate the lungs where the virus persists with no detectable infection in epithelial cells. After glucocorticoid treatment, the virus is reactivated from neural fibers and reinfects epithelial cells. The findings show that HMPV persists in neural fibers and suggest a mechanism for disease chronicity that has important implications for HMPV disease intervention strategies.
A Dominant-Negative Mutant of rab5 Inhibits Infection of Cells by Foot-and-Mouth Disease Virus: Implications for Virus Entry
Authors: Johns Helen L. , Berryman Stephen , Monaghan Paul , Belsham Graham J. , Jackson Terry 
Published: April 2009

ABSTRACT
Foot-and-mouth disease virus (FMDV) can use a number of different integrins (αvβ1, αvβ3, αvβ6, and αvβ8) as receptors to initiate infection. Infection mediated by αvβ6 is known to occur by clathrin-mediated endocytosis and is dependent on the acidic pH within endosomes. On internalization, virus is detected rapidly in early endosomes (EE) and subsequently in perinuclear recycling endosomes (PNRE), but not in late endosomal compartments. Due to the extreme sensitivity of FMDV to acidic pH, it is thought that EE can provide a pH low enough for infection to occur; however, definitive proof that infection takes place from within these compartments is still lacking. Here we have investigated the intracellular transport steps required for FMDV infection of IBRS-2 cells, which express αvβ8 as their FMDV receptor. These experiments confirmed that FMDV infection mediated by αvβ8 is also dependent on clathrin-mediate endocytosis and an acidic pH within endosomes. Also, the effect on FMDV infection of dominant-negative (DN) mutants of cellular rab proteins that regulate endosomal traffic was examined. Expression of DN rab5 reduced the number of FMDV-infected cells by 80%, while expression of DN rab4 or DN rab7 had virtually no effect on infection. Expression of DN rab11 inhibited infection by FMDV, albeit to a small extent (∼35%). These results demonstrate that FMDV infection takes place predominantly from within EE and does not require virus trafficking to the late endosomal compartments. However, our results suggest that infection may not be exclusive to EE and that a small amount of infection could occur from within PNRE.
Primary Human Immunodeficiency Virus Type 1-Specific CD8+T-Cell Responses Induced by Myeloid Dendritic Cells
Authors: Colleton Bonnie A. , Huang Xiao-Li , Melhem Nada M. , Fan Zheng , Borowski Luann , Rappocciolo Giovanna , Rinaldo Charles R. 
Published: April 2009

ABSTRACTInduction of an antigenically broad and vigorous primary T-cell immune response by myeloid dendritic cells (DC) in blood and tissues could be important for an effective prophylactic or therapeutic vaccine to human immunodeficiency virus type 1 (HIV-1). Here we show that a primary CD8+T-cell response can be induced by HIV-1 peptide-loaded DC derived from blood monocytes of HIV-1-negative adults and neonates (moDC) and by Langerhans cells (LC) and interstitial, dermal-intestinal DC (idDC) derived from CD34+stem cells of neonatal cord blood. Optimal priming of single-cell gamma interferon (IFN-γ) production by CD8+T cells required CD4+T cells and was broadly directed to multiple regions of Gag, Env, and Nef that corresponded to known and predicted major histocompatibility complex class I epitopes. Polyfunctional CD8+T-cell responses, defined as single-cell production of more than one cytokine (IFN-γ, interleukin 2, or tumor necrosis factor alpha), chemokine (macrophage inhibitory factor 1β), or cytotoxic degranulation marker CD107a, were primed by moDC, LC, and idDC to HIV-1 Gag and reverse transcriptase epitopes, as well as to Epstein-Barr virus and influenza A virus epitopes. Thus, three major types of blood and tissue myeloid DC targeted by HIV-1, i.e., moDC, LC, and idDC, can prime multispecific, polyfunctional CD8+T-cell responses to HIV-1 and other viral antigens.
Identification of a Novel Determinant for Membrane Association in Hepatitis C Virus Nonstructural Protein 4B
Authors: Gouttenoire Jérôme , Castet Valérie , Montserret Roland , Arora Naveen , Raussens Vincent , Ruysschaert Jean-Marie , Diesis Eric , Blum Hubert E. , Penin François , Moradpour Darius 
Published: April 2009

ABSTRACT
Nonstructural protein 4B (NS4B) plays an essential role in the formation of the hepatitis C virus (HCV) replication complex. It is a relatively poorly characterized integral membrane protein predicted to comprise four transmembrane segments in its central portion. Here, we describe a novel determinant for membrane association represented by amino acids (aa) 40 to 69 in the N-terminal portion of NS4B. This segment was sufficient to target and tightly anchor the green fluorescent protein to cellular membranes, as assessed by fluorescence microscopy as well as membrane extraction and flotation analyses. Circular dichroism and nuclear magnetic resonance structural analyses showed that this segment comprises an amphipathic α-helix extending from aa 42 to 66. Attenuated total reflection infrared spectroscopy and glycosylation acceptor site tagging revealed that this amphipathic α-helix has the potential to traverse the phospholipid bilayer as a transmembrane segment, likely upon oligomerization. Alanine substitution of the fully conserved aromatic residues on the hydrophobic helix side abrogated membrane association of the segment comprising aa 40 to 69 and disrupted the formation of a functional replication complex. These results provide the first atomic resolution structure of an essential membrane-associated determinant of HCV NS4B.
Comparison of the Effectiveness of Six and Two Acupuncture Point Regimens in Osteoarthritis of the Knee: A Randomised Trial
Authors: Taechaarpornkul Wirat , Suvapan Daranee , Theppanom Chaniya , Chanthipwaree Chantima , Chirawatkul Aroon 
Published: April 2009

Background Although substantial data have supported the effectiveness of acupuncture for treating knee osteoarthritis (OA), the number of points used has varied. The objective of this study was to compare the effectiveness of six and two acupuncture points in the treatment of knee OA. Methods A randomised trial of knee OA patients was conducted. Patients were randomly allocated into two groups of 35. The “six point group” received treatment at six acupuncture points, ST35, EX-LE4 ( Neixiyan), ST36, SP9, SP10 and ST34, while the “two point group” received treatment at just the first pair of points, ST35 and EX-LE4. Both groups received twice weekly electroacupuncture on 10 occasions. Electrical stimulation was carried out at low-frequency of 3 Hz to all points, with the intensity as high as tolerable. Both groups were allowed to take a 200 mg celecoxib capsule per day for intolerable pain. Patients were assessed at baseline, week 5, week 9 and week 13, using a Thai language version of the Western Ontario and McMaster Osteoarthritis Index (WOMAC). Global assessment of change after 10 treatments was also recorded. Results Acupuncture at both six and two acupuncture points was associated with significant improvement. Mean total WOMAC score at weeks 5 and 13 of patients in both groups showed no significant difference statistically (p = 0.75 and p = 0.51). Moreover, the number of celecoxib capsules taken, global assessment of global change and body weight change of both groups also showed no statistical difference. Conclusion This evidence suggests that electroacupuncture to two local points may be sufficient to treat knee OA, but in view of some limitations to this study further research is necessary before this can be stated conclusively.
Gatekeepers and the Gateway – a Mixed-Methods Inquiry into Practitioners' Referral Behaviour to the Gateway Clinic
Authors: Unwin Jason , Peters David 
Published: April 2009

Background The Gateway Clinic is a specialist NHS service in South London providing acupuncture and Chinese medicine. The clinic receives a diverse range of referrals from an expanding population of local general practitioners (GPs). Objectives This study explores the referral behaviour of GPs and other primary care users of the Gateway Clinic. Methods A pragmatic mixed-methods approach was used, combining mapping of GPs’ usage of the Gateway as taken from the clinic's database between the years 2000 and 2005, a questionnaire survey of the referring primary care practitioners in 2005, and data, including a thematic analysis of the interview transcripts from semi-structured interviews of a purposive sample of those GPs who refer. Results Doctors in the community refer a wide range of health problems to the Gateway Clinic. The most common referrals were musculoskeletal conditions (38–44%), general and unspecified conditions (11–14%), blood and immune system conditions (6–12%), psychological conditions (8–13%), neurological conditions (6–9%), gynaecological conditions (5–6%) and digestive conditions (5–6%). The health problems referred by local healthcare practitioners correlate strongly with the patient profile seen in private acupuncture practice. Between 2000 and 2005 numbers of healthcare practitioners increased by 218% resulting in a 300% increase in patient referrals made to the clinic; the mean frequency of referral per practitioner increased by 71%. Analysis of the qualitative data suggests that positive clinical experience encourages GPs to increase the range of conditions they refer leading to the development of informal referral guidelines. Conclusion The Gateway Clinic has become an increasingly popular referral resource. The influences that drive referral to the clinic are multiple and follow “tacit guidelines”. GPs select patients on the basis of their individual clinical experience, informed by positive patient feedback and often only after more conventional medical treatment options have been exhausted.
Pilot Study to Assess the Credibility of Acupuncture in Acute Exacerbations of Chronic Obstructive Pulmonary Disease
Authors: Whale Claudia A , MacLaran Sarah J A , Whale Christopher I , Barnett Mandy 
Published: April 2009

Background Exacerbation of chronic obstructive pulmonary disease (COPD) is a common reason for hospital admission, and adjunctive non-pharmacological treatments would be welcomed. A pilot study was undertaken to assess the feasibility of conducting a study of acupuncture during an acute exacerbation of COPD. We also examined the credibility of a sham device in this setting and assessed the effect of acupuncture on breathlessness and anxiety. Methods A prospective, randomised, patient- and assessor-blinded, sham controlled study was conducted on three consecutive days in a district general hospital. Credibility of both acupuncture and the Park sham device were assessed using the Borkovec and Nau questionnaire. Dyspnoea was measured on the modified Borg score and a 10 cm visual analogue scale, while anxiety was measured on a 10 cm visual analogue scale. Results 11 patients were recruited and nine completed the study. There were no adverse events with either intervention. Acupuncture was well tolerated and credibility scores were similar before and after real and sham acupuncture. Symptoms improved after both treatments, with no significant difference between groups. Conclusion In this pilot study acupuncture was well tolerated by subjects experiencing an acute exacerbation of COPD. Acupuncture treatment and the Park sham device were both credible. Although recruitment was slow, a further trial with a larger sample size is feasible and recommended.