bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of an Experimental Vaccine for Bovine Respiratory Syncytial Virus
Authors: Hägglund Sara , Hu Kefei , Blodörn Krister , Makabi-Panzu Boby , Gaillard Anne-Laure , Ellencrona Karin , Chevret Didier , Hellman Lars , Bengtsson Karin Lövgren , Riffault Sabine , Taylor Geraldine , Valarcher Jean François , Eléouët Jean-François
Published: May 2014

Bovine respiratory syncytial virus (BRSV) and human respiratory syncytial virus (HRSV) are major causes of respiratory disease in calves and children, respectively, and are priorities for vaccine development. We previously demonstrated that an experimental vaccine, BRSV-immunostimulating complex (ISCOM), is effective in calves with maternal antibodies. The present study focuses on the antigenic characterization of this vaccine for the design of new-generation subunit vaccines. The results of our study confirmed the presence of membrane glycoprotein (G), fusion glycoprotein (F), and nucleoprotein (N) proteins in the ISCOMs, and this knowledge was extended by the identification of matrix (M), M2-1, phosphoprotein (P), small hydrophobic protein (SH) and of cellular membrane proteins, such as the integrins αVβ1, αVβ3, and α3β1. The quantity of the major protein F was 4- to 5-fold greater than that of N (∼77 μg versus ∼17 μg/calf dose), whereas G, M, M2-1, P, and SH were likely present in smaller amounts. The polymerase (L), M2-2, nonstructural 1 (NS1), and NS2 proteins were not detected, suggesting that they are not essential for protection. Sera from the BRSV-ISCOM-immunized calves contained high titers of IgG antibody specific for F, G, N, and SH. Antibody responses against M and P were not detected; however, this does not exclude their role in protective T-cell responses. The absence of immunopathological effects of the cellular proteins, such as integrins, needs to be further confirmed, and their possible contribution to adjuvant functions requires elucidation. This work suggests that a combination of several surface and internal proteins should be included in subunit RSV vaccines and identifies absent proteins as potential candidates for differentiating infected from vaccinated animals.
Molecular Tools for Diagnosis of Visceral Leishmaniasis: Systematic Review and Meta-Analysis of Diagnostic Test Accuracy
Authors: de Ruiter C. M. , van der Veer C. , Leeflang M. M. G. , Deborggraeve S. , Lucas C. , Adams E. R.
Published: May 2014

Molecular methods have been proposed as highly sensitive tools for the detection of
Leishmania
parasites in visceral leishmaniasis (VL) patients. Here, we evaluate the diagnostic accuracy of these tools in a meta-analysis of the published literature. The selection criteria were original studies that evaluate the sensitivities and specificities of molecular tests for diagnosis of VL, adequate classification of study participants, and the absolute numbers of true positives and negatives derivable from the data presented. Forty studies met the selection criteria, including PCR, real-time PCR, nucleic acid sequence-based amplification (NASBA), and loop-mediated isothermal amplification (LAMP). The sensitivities of the individual studies ranged from 29 to 100%, and the specificities ranged from 25 to 100%. The pooled sensitivity of PCR in whole blood was 93.1% (95% confidence interval [CI], 90.0 to 95.2), and the specificity was 95.6% (95% CI, 87.0 to 98.6). The specificity was significantly lower in consecutive studies, at 63.3% (95% CI, 53.9 to 71.8), due either to true-positive patients not being identified by parasitological methods or to the number of asymptomatic carriers in areas of endemicity. PCR for patients with HIV-VL coinfection showed high diagnostic accuracy in buffy coat and bone marrow, ranging from 93.1 to 96.9%. Molecular tools are highly sensitive assays for
Leishmania
detection and may contribute as an additional test in the algorithm, together with a clear clinical case definition. We observed wide variety in reference standards and study designs and now recommend consecutively designed studies.
Genetic Structure of Mycobacterium avium subsp. paratuberculosis Population in Cattle Herds in Quebec as Revealed by Using a Combination of Multilocus Genomic Analyses
Authors: Sohal Jagdip Singh , Arsenault Julie , Labrecque Olivia , Fairbrother Julie-Hélène , Roy Jean-Philippe , Fecteau Gilles , L'Homme Yvan
Published: May 2014

Mycobacterium avium
subsp.
paratuberculosis
is the etiological agent of paratuberculosis, a granulomatous enteritis affecting a wide range of domestic and wild ruminants worldwide. A variety of molecular typing tools are used to distinguish
M. avium
subsp.
paratuberculosis
strains, contributing to a better understanding of
M. avium
subsp.
paratuberculosis
epidemiology. In the present study, PCR-based typing methods, including mycobacterial interspersed repetitive units/variable-number tandem repeats (MIRU-VNTR) and small sequence repeats (SSR) in addition to IS1311 PCR-restriction enzyme analysis (PCR-REA), were used to investigate the genetic heterogeneity of 200
M. avium
subsp.
paratuberculosis
strains from dairy herds located in the province of Quebec, Canada. The majority of strains were of the “cattle type,” or type II, although 3 strains were of the “bison type.” A total of 38 genotypes, including a novel one, were identified using a combination of 17 genetic markers, which generated a Simpson's index of genetic diversity of 0.876. Additional analyses revealed no differences in genetic diversity between environmental and individual strains. Of note, a spatial and spatiotemporal cluster was evidenced regarding the distribution of one of the most common genotypes. The population had an overall homogeneous genetic structure, although a few strains stemmed out of the consensus cluster, including the bison-type strains. The genetic structure of
M. avium
subsp.
paratuberculosis
populations within most herds suggested intraherd dissemination and microevolution, although evidence of interherd contamination was also revealed. The level of genetic diversity obtained by combining MIRU-VNTR and SSR markers shows a promising avenue for molecular epidemiology investigations of
M. avium
subsp.
paratuberculosis
transmission patterns.
Characterization of Enterohemorrhagic Escherichia coli O111 and O157 Strains Isolated from Outbreak Patients in Japan
Authors: Watahiki Masanori , Isobe Junko , Kimata Keiko , Shima Tomoko , Kanatani Jun-ichi , Shimizu Miwako , Nagata Akihiro , Kawakami Keiko , Yamada Mikiko , Izumiya Hidemasa , Iyoda Sunao , Morita-Ishihara Tomoko , Mitobe Jiro , Terajima Jun , Ohnishi Makoto , Sata Tetsutaro
Published: May 2014

In April and May 2011, there was a serious food-poisoning outbreak in Japan caused by enterohemorrhagic
Escherichia coli
(EHEC) strains O111:H8 and O157:H7 from raw beef dishes at branches of a barbecue restaurant. This outbreak involved 181 infected patients, including 34 hemolytic-uremic syndrome (HUS) cases (19%). Among the 34 HUS patients, 21 developed acute encephalopathy (AE) and 5 died. Patient stool specimens yielded
E. coli
O111 and O157 strains. We also detected both EHEC O111
stx
2
and
stx
-negative
E. coli
O111 strains in a stock of meat block from the restaurant. Pulsed-field gel electrophoresis (PFGE) and multilocus variable-number tandem-repeat analysis (MLVA) showed that the
stx
-negative
E. coli
O111 isolates were closely related to EHEC O111
stx
2
isolates. Although the EHEC O157 strains had diverse
stx
gene profiles (
stx
1
,
stx
2
, and
stx
1
stx
2
), the PFGE and MLVA analyses indicated that these isolates originated from a single clone. Deletion of the Stx2-converting prophage from the EHEC O111
stx
2
isolates was frequently observed during
in vitro
growth, suggesting that strain conversion from an EHEC O111
stx
2
to an
stx
-negative strain may have occurred during infection.
First Report of Nocardia beijingensis Infection in an Immunocompetent Host in the United States
Authors: Crozier Jennifer A. , Andhavarapu Swati , Brumble Lisa M. , Sher Taimur
Published: May 2014

Here we describe the first reported case of
Nocardia beijingensis
infection in the United States, made rarer by its presence in an immunocompetent patient.
Paravertebral Mushroom: Identification of a Novel Species of Phellinus as a Human Pathogen in Chronic Granulomatous Disease
Authors: De Ravin Suk See , Parta Mark , Sutton Deanna A. , Wickes Brian L. , Thompson Elizabeth H. , Wiederhold Nathan P. , Nakasone Karen K. , Alimchandani Meghna , OConnell Amy , Notarangelo Luigi , Kang Elizabeth , Malech Harry L. , Zelazny Adrian M.
Published: May 2014

We describe a case of paravertebral abscess caused by a
Phellinus
sp. in a boy with chronic granulomatous disease. Sequence-based identification of this mold, a new agent of disease, suggests a close relation to
Phellinus umbrinellus
.
Prevalence and Detection of Mixed-Population Enterococcal Bacteremia
Authors: Cárdenas Ana María , Andreacchio Kathleen A. , Edelstein Paul H.
Published: May 2014

Mixed-population (heterogeneous) enterococcal bacteremia (MEB) is rarely reported. Based on one occasion in which Vitek2 missed a vancomycin-resistant subpopulation isolated from a patient, we developed a simple method to detect this subpopulation and determined MEB frequency. The four patients presented here had either
Enterococcus faecium
or
Enterococcus faecalis
bacteremia caused by both vancomycin-resistant enterococci (VRE) and vancomycin-susceptible enterococci (VSE). No prior common antibiotic therapy was observed, and bacteremia resolved with daptomycin, gentamicin, and/or linezolid treatment. In two cases, VRE presence was missed by Vitek2. To detect the VRE subpopulation, tryptic soy broth was inoculated from positive blood cultures and a saline suspension was inoculated to a vancomycin (6-μg/ml) (V6) plate. Two isolates from each patient were studied further. Relatedness was assessed by multilocus sequence typing, fitness was evaluated by growth curve and competition assays, and
vanA
presence was determined by PCR. MEB represented ∼5% of all enterococcal bacteremias. All VRE subpopulations grew on V6 plates but were missed in two instances by Vitek2. VRE and VSE isolates from each patient were closely related and did not differ in overall fitness. All four VRE isolates and 2/4 VSE isolates were
vanA
positive. MEBs occur regardless of prior antimicrobial therapy, are relatively common in our hospital, and are important to detect. As far as we know, this study is the first to report heterogeneous
E. faecalis
bacteremia. There is a simple method to detect VRE subpopulations that may be missed by Vitek2.
Evolution of Recombinant Lymphocytic Choriomeningitis Virus/Lassa Virus
In Vivo
Highlights the Importance of the GPC Cytosolic Tail in Viral Fitness
Authors: Sommerstein Rami , Ramos da Palma Joel , Ölschläger Stephan , Bergthaler Andreas , Barba Leticia , Lee Boris P.-L. , Pasquato Antonella , Flatz Lukas
Published: May 2014

A key characteristic of arenaviruses is their ability to establish persistent infection in their natural host. Different factors like host age, viral dose strain, and route of infection may contribute to the establishment of persistence. However, the molecular mechanisms governing persistence are not fully understood. Here, we describe gain-of-function mutations of lymphocytic choriomeningitis virus (LCMV) expressing Lassa virus (LASV) GP, which can prolong viremia in mice depending on the sequences in the GP-2 cytoplasmic tail. The initial mutant variant (rLCMV/LASV mut GP) carried a point mutation in the cytosolic tail of the LASV glycoprotein GP corresponding to a K461G substitution. Unlike what occurred with the original rLCMV/LASV wild-type (wt) GP, infection of C57BL/6 mice with the mutated recombinant virus led to a detectable viremia of 2 weeks' duration. Further replacement of the entire sequence of the cytosolic tail from LASV to LCMV GP resulted in increased viral titers and delayed clearance of the viruses. Biosynthesis and cell surface localization of LASV wt and mut GPs were comparable.

IMPORTANCE
Starting from an emerging virus in a wild-type mouse, we engineered a panel of chimeric Lassa/lymphocytic choriomeningitis viruses. Mutants carrying a viral envelope with the cytosolic tail from the closely related mouse-adapted LCMV were able to achieve a productive viral infection lasting up to 27 days in wild-type mice. Biochemical assays showed a comparable biosynthesis and cell surface localization of LASV wt and mut GPs. These recombinant chimeric viruses could allow the study of immune responses and antivirals targeting the LASV GP.
A Genome-Wide Small Interfering RNA Screen Identifies Host Factors Required for Vesicular Stomatitis Virus Infection
Authors: Lee Amy Si-Ying , Burdeinick-Kerr Rebeca , Whelan Sean P. J.
Published: May 2014

Viruses are dependent on their host cells for replication and thus have evolved in intimate association with them. The identification of host factors required for viral infection has led to advances in both viral and cellular biology. Vesicular stomatitis virus (VSV), a negative-sense RNA virus, replicates in all eukaryotic cells in culture, suggesting that the host requirements for its replication are ubiquitous. In this study, we performed a genome-wide small interfering RNA screen of human cells in culture and identified multiple cellular genes that influence the entry and replication of VSV. From a list of >300 genes, we selected the most promising candidates to perform further analysis to assign their functions to either the entry or intracellular replication step of infection. We implicate 3 new factors in VSV entry and 20 new factors in viral gene expression. These proteins have diverse cellular roles, including
S
-adenosylmethionine synthesis, respiration, and host translation machinery, underscoring the intimate relationship between VSV and the host cell. Together, these results provide a curated list of genes required for VSV replication.

IMPORTANCE
Replication of vesicular stomatitis virus (VSV) has long served as a model for understanding host-virus interactions and neuropathogenesis. We performed a genome-wide analysis of host factors and revealed genes critical for viral replication, including some involved in vesicular trafficking, cell cycling, and protein modification. Our results provide an enriched list of host factors that are required for specific stages of VSV entry and gene expression. This study may also potentially expand the repertoire of targets for antiviral therapy against negative-strand RNA viruses.
Paramyxovirus V Protein Interaction with the Antiviral Sensor LGP2 Disrupts MDA5 Signaling Enhancement but Is Not Relevant to LGP2-Mediated RLR Signaling Inhibition
Authors: Rodriguez Kenny R. , Horvath Curt M.
Published: May 2014

The interferon antiviral system is a primary barrier to virus replication triggered upon recognition of nonself RNAs by the cytoplasmic sensors encoded by retinoic acid-inducible gene I (RIG-I), melanoma differentiation-associated gene 5 (MDA5), and laboratory of genetics and physiology gene 2 (LGP2). Paramyxovirus V proteins are interferon antagonists that can selectively interact with MDA5 and LGP2 through contact with a discrete helicase domain region. Interaction with MDA5, an activator of antiviral signaling, disrupts interferon gene expression and antiviral responses. LGP2 has more diverse reported roles as both a coactivator of MDA5 and a negative regulator of both RIG-I and MDA5. This functional dichotomy, along with the concurrent interference with both cellular targets, has made it difficult to assess the unique consequences of V protein interaction with LGP2. To directly evaluate the impact of LGP2 interference, MDA5 and LGP2 variants unable to be recognized by measles virus and parainfluenza virus 5 (PIV5) V proteins were tested in signaling assays. Results indicate that interaction with LGP2 specifically prevents coactivation of MDA5 signaling and that LGP2's negative regulatory capacity was not affected. V proteins only partially antagonize RIG-I at high concentrations, and their expression had no additive effects on LGP2-mediated negative regulation. However, conversion of RIG-I to a direct V protein target was accomplished by only two amino acid substitutions that allowed both V protein interaction and efficient interference. These results clarify the unique consequences of MDA5 and LGP2 interference by paramyxovirus V proteins and help resolve the distinct roles of LGP2 in both activation and inhibition of antiviral signal transduction.

IMPORTANCE
Paramyxovirus V proteins interact with two innate immune receptors, MDA5 and LGP2, but not RIG-I. V proteins prevent MDA5 from signaling to the beta interferon promoter, but the consequences of LGP2 targeting are poorly understood. As the V protein targets MDA5 and LGP2 simultaneously, and LGP2 is both a positive and negative regulator of both MDA5 and RIG-I, it has been difficult to evaluate the specific advantages conferred by LGP2 targeting. Experiments with V-insensitive proteins revealed that the primary outcome of LGP2 interference is suppression of its ability to synergize with MDA5. LGP2's negative regulation of MDA5 and RIG-I remains intact irrespective of V protein interaction. Complementary experiments demonstrate that RIG-I can be converted to V protein sensitivity by two amino acid substitutions. These findings clarify the functions of LGP2 as a positive regulator of MDA5 signaling, demonstrate the basis for V-mediated LGP2 targeting, and broaden our understanding of paramyxovirus-host interactions.
Glutamic Acid at Residue 125 of the prM Helix Domain Interacts with Positively Charged Amino Acids in E Protein Domain II for Japanese Encephalitis Virus-Like-Particle Production
Authors: Peng Jia-Guan , Wu Suh-Chin
Published: May 2014

Interaction between E and prM proteins in flavivirus-infected cells is a major factor for virus-like particle (VLP) production. The prM helical (prM-H) domain is topologically close to and may interact with domain II of the E protein (EDII). In this study, we investigated prM-H domain amino acid residues facing Japanese encephalitis virus EDII using site-directed mutagenesis to determine their roles in prM-E interaction and VLP production. Our results indicate that negatively charged prM-E125 residue at the prM-H domain affected VLP production via one or more interactions with positively charged E-K93 and E-H246 residues at EDII. Exchanges of oppositely charged residue side chains at prM-E125/E-K93 and prM-E125/E-H246 are recoverable for VLP production. The prM-E125 and E-H246 residues are conserved and that the positive charge of the E-K93 residue is preserved in different flavivirus groups. These findings suggest that the electrostatic attractions of prM-E125, E-K93, and E-H246 residues are important to flavivirus VLP production and that inhibiting these interactions is a potential strategy for blocking flavivirus infections.

IMPORTANCE
Molecular interaction between E and prM proteins of Japanese encephalitis virus is a major driving force for virus-like particle (VLP) production. The current high-resolution structures available for prM-E complexes do not include the membrane proximal stem region of prM. The prM stem region contains an N-terminal loop and a helix domain (prM-H). Since the prM-H domain is topologically close to domain II of the E protein (EDII), this study was to determine molecular interactions between the prM-H domain and EDII. We found that the molecular interactions between prM-E125 residue and positively charged E-K93 and E-H246 residues at EDII are critical for VLP production. More importantly, the prM-E125 and E-H246 residues are conserved and the positive charge of the E-K93 residue is preserved in different flavivirus groups. Our findings help refine the structure and molecular interactions on the flavivirus surface and reveal a potential strategy for blocking flavivirus infections by inhibiting these electrostatic interactions.
Endoplasmic Reticulum Stress Links Hepatitis C Virus RNA Replication to Wild-Type PGC-1α/Liver-Specific PGC-1α Upregulation
Authors: Yao Wenxia , Cai Hua , Li Xinlei , Li Ting , Hu Longbo , Peng Tao
Published: May 2014

Hepatitis C virus (HCV) causes not only severe liver problems but also extrahepatic manifestations, such as insulin resistance (IR). Wild-type peroxisome proliferator-activated receptor gamma coactivator 1 alpha (WT-PGC-1α) is essential in hepatic gluconeogenesis and has recently been demonstrated to link HCV infection to hepatic insulin resistance (IR). A recent study has characterized a novel human liver-specific PGC-1α (L-PGC-1α) transcript, which is proposed to reflect human adaption to more complex pathways. However, the effect of HCV infection on L-PGC-1α expression and the mechanism by which HCV modulates WT-PGC-1α/L-PGC-1α remain unclear. In this study, we showed that HCV infection upregulated both WT-PGC-1α and L-PGC-1α, which further promoted HCV production. The upregulation of both PGC-1α isoforms depended on HCV RNA replication. By using promoter-luciferase reporters, kinase inhibitors, and dominant negative mutants, we further observed that the HCV-induced upregulation of WT-PGC-1α was mediated by the phosphorylation of cyclic AMP (cAMP)-responsive element-binding protein (CREB), whereas that of L-PGC-1α was mediated by CREB phosphorylation and forkhead box O1 dephosphorylation. Moreover, HCV infection induced endoplasmic reticulum (ER) stress, and pharmacological induction of ER stress upregulated WT-PGC-1α/L-PGC-1α and phosphorylated CREB. In contrast, pharmacological inhibition of HCV-induced ER stress impaired WT-PGC-1α/L-PGC-1α upregulation along with decreased phosphorylated CREB. The correlation of hepatic mPGC-1α with ER stress was further confirmed in mice. Overall, HCV infection upregulates both WT-PGC-1α and L-PGC-1α through an ER stress-mediated, phosphorylated CREB-dependent pathway, and both PGC-1α isoforms promote HCV production in turn.

IMPORTANCE
HCV causes not only severe liver problems but also extrahepatic manifestations, such as insulin resistance (IR). As a key regulator in energy metabolism, wild-type PGC-1α (WT-PGC-1α), has recently been demonstrated to link HCV infection to hepatic IR. A recent study has characterized a novel human liver-specific PGC-1α (L-PGC-1α), which reflects human adaption to more complex pathways. However, the effect of HCV infection on L-PGC-1α expression and the mechanism by which HCV regulates WT-PGC-1α/L-PGC-1α remain unclear. In this study, we showed that HCV infection upregulated both WT-PGC-1α and L-PGC-1α, which further promoted HCV production. WT-PGC-1α upregulation was mediated by CREB phosphorylation, whereas L-PGC-1α upregulation was mediated by CREB phosphorylation and FoxO1 dephosphorylation. HCV-induced ER stress mediated WT-PGC-1α/L-PGC-1α upregulation and CREB phosphorylation. Overall, this study provides new insights into the mechanism by which HCV upregulates WT-PGC-1α/L-PGC-1α and highlights the novel intervention of HCV-ER stress-PGC-1α signaling for HCV therapy and HCV-induced IR therapy.
Faster Replication and Higher Expression Levels of Viral Glycoproteins Give the Vesicular Stomatitis Virus/Measles Virus Hybrid VSV-FH a Growth Advantage over Measles Virus
Authors: Ayala-Breton Camilo , Russell Luke O. J. , Russell Stephen J. , Peng Kah-Whye
Published: May 2014

VSV-FH is a hybrid vesicular stomatitis virus (VSV) with a deletion of its G glycoprotein and encoding the measles virus (MV) fusion (F) and hemagglutinin (H) envelope glycoproteins. VSV-FH infects cells expressing MV receptors and is fusogenic and effective against myeloma xenografts in mice. We evaluated the fusogenic activities of MV and VSV-FH in relationship to the density of receptor on the target cell surface and the kinetics of F and H expression in infected cells. Using a panel of cells expressing increasing numbers of the MV receptor CD46, we evaluated syncytium size in MV- or VSV-FH-infected cells. VSV-FH is not fusogenic at low CD46 density but requires less CD46 for syncytium formation than MV. The size of each syncytium is larger in VSV-FH-infected cells at a specific CD46 density. While syncytium size reached a plateau and did not increase further in MV-infected CHO cells expressing ≥4,620 CD46 copies/cell, there was a corresponding increase in syncytium size with increases in CD46 levels in VSV-FH-infected CD46-expressing CHO (CHO-CD46) cells. Further analysis in VSV-FH-infected cell lines shows earlier and higher expression of F and H mRNAs and protein. However, VSV-FH cytotoxic activity was reduced by pretreatment of the cells with type I interferon. In contrast, the cytopathic effects are not affected in MV-infected cells. In summary, VSV-FH has significant advantages over MV as an oncolytic virus due to its higher viral yield, faster replication kinetics, and larger fusogenic capabilities but should be used in cancer types with defective interferon signaling pathways.

IMPORTANCE
We studied the cytotoxic activity of a vesicular stomatitis/measles hybrid virus (VSV-FH), which is superior to that of measles virus (MV), in different cancer cell lines. We determined that viral RNA and protein were produced faster and in higher quantities in VSV-FH-infected cells. This resulted in the formation of larger syncytia, higher production of infectious particles, and a more potent cytopathic effect in permissive cells. Importantly, VSV-FH, similar to MV, can discriminate between low- and high-expressing CD46 cells, a phenotype important for cancer therapy as the virus will be able to preferentially infect cancer cells that overexpress CD46 over low-CD46-expressing normal cells.
Increased Mucosal CD4
+
T Cell Activation in Rhesus Macaques following Vaccination with an Adenoviral Vector
Authors: Bukh Irene , Calcedo Roberto , Roy Soumitra , Carnathan Diane G. , Grant Rebecca , Qin Qiuyue , Boyd Surina , Ratcliffe Sarah J. , Veeder Christin L. , Bellamy Scarlett L. , Betts Michael R. , Wilson James M.
Published: May 2014

The possibility that vaccination with adenovirus (AdV) vectors increased mucosal T cell activation remains a central hypothesis to explain the potential enhancement of HIV acquisition within the Step trial. Modeling this within rhesus macaques is complicated because human adenoviruses, including human adenovirus type 5 (HAdV-5), are not endogenous to macaques. Here, we tested whether vaccination with a rhesus macaque-derived adenoviral vector (simian adenovirus 7 [SAdV-7]) enhances mucosal T cell activation within rhesus macaques. Following intramuscular SAdV-7 vaccination, we observed a pronounced increase in SAdV-7-specific CD4
+
T cell responses in peripheral blood and, more dramatically, in rectal mucosa tissue. Vaccination also induced a significant increase in the frequency of activated memory CD4
+
T cells in SAdV-7- and HAdV-5-vaccinated animals in the rectal mucosa but not in peripheral blood. These fluctuations within the rectal mucosa were also associated with a pronounced decrease in the relative frequency of naive resting CD4
+
T cells. Together, these results indicate that peripheral vaccination with an AdV vector can increase the activation of mucosal CD4
+
T cells, potentially providing an experimental model to further evaluate the role of host-vector interactions in increased HIV acquisition after AdV vector vaccination.

IMPORTANCE
The possibility that vaccination with a human adenovirus 5 vector increased mucosal T cell activation remains a central hypothesis to explain the potential enhancement of human immunodeficiency virus (HIV) acquisition within the Step trial. In this study, we tested whether vaccination with a rhesus macaque-derived adenoviral vector in rhesus macaques enhances mucosal CD4
+
T cell activation, the main cell target of simian immunodeficiency virus (SIV)/HIV. The results showed that vaccination with an adenoviral vector indeed increases activation of mucosal CD4
+
T cells and potentially increases susceptibility to SIV infection.
Severe flaccid paraparesis following spinal anaesthesia: asine materiaoccurrence
Authors: Vyshka Gentian , Vacchiano Giuseppe
Published: May 2014

We report the case of a 45-year-old female patient who underwent spinal anaesthesia for a minor urological intervention, trying to correct her stress urinary incontinence. Hyperbaric bupivacaine was intrathecally injected and the surgical intervention ended uneventfully. Six hours after the operation a severe motor deficit of lower limbs was noted. The situation persisted and neurologists repeatedly checked on the patient; imaging of the lumbar spine was made but no pathological signal was registered. Somatosensory evoked potentials of the lower limbs showed bilaterally delayed latencies of the cortical components, with prolonging of interpeak latencies, suggestive of a suffering of the long somatosensory spinal tracts. Unable initially to walk or stand in erect position with almost no movements in both lower limbs, the patient was admitted in a rehabilitation facility and treated there for 4 months, with her condition substantially improving during this period.
Infant case of lysosomal acid lipase deficiency: Wolman's disease
Authors: Sadhukhan Meghmala , Saha Amit , Vara Roshni , Bhaduri Bim
Published: May 2014

Lysosomal acid lipase (LAL) deficiency is a rare autosomal recessive disorder which causes two distinct clinical phenotypes: Wolman's disease and cholesterol ester storage disease. LAL hydrolyses LDL-derived triglycerides and cholesterol esters to glycerol or cholesterol and free fatty acids. Its deficiency leads to accumulation of intracellular triglycerides and/or cholesterol esters. In early onset LAL deficiency, clinical manifestations start in the first few weeks of life with persistent vomiting, failure to thrive, hepatosplenomegaly, liver dysfunction and hepatic failure. Adrenal calcification is a striking feature but is present in only about 50% of cases. We report a case of an infant presenting with vomiting, diarrhoea, hepatosplenomegaly and poor weight gain that was subsequently diagnosed as Wolman's disease. He was entered into a clinical trial for LAL replacement therapy. This case reinforces that early onset LAL deficiency should be considered in a baby presenting with failure to thrive, gastrointestinal symptoms and hepatosplenomegaly.
Antenatal mild hydronephrosis with subsequent polyp of the upper ureter in a child presenting with recurrent Dietl's crisis
Authors: Patel Ramnik V , Johal Navroop , Evans Kathryn , Mushtaq Imran
Published: May 2014

We report a 7-year-old boy who had antenatally diagnosed and postnatally confirmed asymptomatic right congenital hydronephrosis secondary to ureteropelvic junction obstruction with the anteroposterior diameter ranging from 7 to 15 mm on serial ultrasound scans till the age of 6 years. He then presented with recurrent attacks of Dietl's crisis almost every month in the seventh year of his life and eluded diagnosis for over a year at various national hospitals, and ended up with an international referral to us. Investigations of the acute attacks clinched the diagnosis of ureteral polyp causing intermittent crisis and he underwent segmental resection and reconstruction in the form of dismembered Anderson-Hyne pyeloplasty with good recovery. Our case revealed that prenatally detected hydronephrosis may worsen after spontaneous postnatal improvement and a polyp acting as a flip valve may produce intermittent hydronephrosis and symptoms later in life. The child should undergo urgent investigations during acute symptoms.
Pentology of Fallot's presenting with complete heart block
Authors: Sethi Rishi , Khare Rashi , Pradhan Akshyaya , Narain Varun Shankar
Published: May 2014

A 17-year-old male patient presented with cyanosis, repeated squatting since childhood and haemoptysis since the past 1 month. He had central cyanosis with clubbing. Cardiovasular examination revealed ejection systolic murmur in the pulmonary area with single S2. ECG showed right ventricular hypertrophy (RVH) with right atrial enlargement and first-degree heart block. Two-dimensional echo showed ventricular septal defect, overriding aorta, RVH, right ventricular enlargement (RVE) and right atrial enlargement with infundibular and valvular pulmonary stenosis and 1.9 cm ostium secondum atrial septal defect. There was no evidence of atrioventricular canal defect. The patient was diagnosed with pentology of Fallots. Follow-up ECG showed complete heart block (CHB) that again reverted to first-degree heart block. A diagnosis of pentology of Fallot with intermittent CHB was made with an awake heart rate of 50/min. This case report shows association of CHB with tetralogy of Fallot.
Improving Patient Safety of Acute Care Lumbar Punctures
Authors: Ormerod Victoria
Published: May 2014

Lumbar puncture (LP) is a common invasive procedure in the acute medical setting but is not without its risks and complications, making best clinical practice and correct documentation important for patient safety. Previous audit revealed poor levels of consistency in technique and documentation in the acute medical setting, highlighting it as an area for improvement.

This project aims to identify current levels of documentation and improve upon these through the departmental education and the introduction of a documentation tool to create a safer clinical environment for LPs. Gold standards in clinical practice were identified through literature review and national guidelines, establishing 15 key parameters as essential areas for documentation.

Patient notes were retrospectively analysed after LP over a two month period to identify levels of documentation in these areas, and the clinical technique used. Results of this initial audit were presented to the department along with an education session regarding current evidence based best practice for LPs and the important aspects relating to patient safety. A documentation tool was also introduced. A re-audit was then performed of the same parameters and assessing the use of the documentation tool.

Results showed a significant increase in overall documentation from 44% up to 95% after intervention, with 85% of cases using the new proforma. We can conclude that the introduction of a documentation tool and departmental education has significantly improved upon LP documentation in the acute medical setting. This is important for both the protection of medical professionals, as well as patient safety and quality of care, and could be implemented in other clinical environments.
Socioeconomic position and the association between anticipated and actual survival in older English adults
Authors: Adams Jean , Stamp Elaine , Nettle Daniel , Milne Eugene M G , Jagger Carol
Published: May 2014

Background

Socioeconomic disadvantage may cause individuals to have lower expectations of longevity and not engage in healthy behaviours because they judge the long-term health benefits of these to be minimal.

We explored demographic, health behaviour, health and socioeconomic correlates of subjectively estimated lifespan (‘anticipated survival’); the ability of anticipated survival to predict actual survival; and whether the predictive ability of anticipated survival differed by other variables, particularly socioeconomic position.

Methods

Data were from wave 1 of the English Longitudinal Study of Ageing. Anticipated survival for up to 25 years was measured on a scale of 0–100. Actual survival was measured over a mean of 6 years, and socioeconomic position using education, household income, occupational class and area deprivation.

Results

Of 10 768 participants, 2255 (21%) died during follow-up. Anticipated survival was positively associated with socioeconomic position, and was greater in women, younger individuals, non-smokers and those who were not widowed, consumed more alcohol, were more physically active, and reported better physical and mental health. After full adjustment, anticipated survival remained positively associated with actual survival. Those reporting low anticipated survival were more likely to die over time than those reporting moderate anticipated survival (HR (95% CIs 1.11 (1.00 to 1.23). The relationship differed significantly by income and age, being strongest in younger individuals and those with higher household income.

Conclusions

Anticipated survival varied with other variables as expected and reflected actual survival. Younger individuals and those with higher household income were better able to identify subtle differences associated with actual survival.
Analysis of hemodynamics and wall mechanics at sites of cerebral aneurysm rupture
Authors: Cebral Juan R , Vazquez Mariano , Sforza Daniel M , Houzeaux Guillaume , Tateshima Satoshi , Scrivano Esteban , Bleise Carlos , Lylyk Pedro , Putman Christopher M
Published: May 2014

Background

It is thought that aneurysms evolve as the result of progressive degradation of the wall in response to abnormal hemodynamics characterized by either high or low wall shear stress (WSS).

Objective

To investigate the effects of these two different hemodynamic pathways in a series of cerebral aneurysms with known rupture sites.

Methods

Nine aneurysms in which the rupture site could be identified in three-dimensional images were analyzed. The WSS distribution was obtained from computational fluid dynamics (CFD) simulations. Internal wall stresses were computed using structural wall models under hemodynamic loads determined by the CFD models. Wall properties (thickness and stiffness) were modulated with the WSS distribution (increased or decreased in regions of high or low WSS) to test possible wall degradation pathways. Rupture probability indices (RPI) were calculated to compare different wall models.

Results

Most rupture sites aligned with the intrasaccular flow stream and downstream of the primary impaction zone. The model that best explained the rupture site (produced higher RPI) in eight of the nine aneurysms (89%) had thinner and stiffer walls in regions of abnormally high WSS. The remaining case (11%) was best explained by a model with thinner and stiffer walls in regions of abnormally low WSS.

Conclusions

Aneurysm rupture seems to be caused by localized degradation and weakening of the wall in response to abnormal hemodynamics. Image-based computational models assuming wall thinning and stiffening in regions of abnormally high WSS were able to explain most of the observed rupture sites.
Mechanism of a novel missense mutation, p.V174M, of the human connexin31 (GJB3) in causing nonsyndromic hearing loss
Authors: Li Tung-Cheng , Kuan Yu-Hsiang , Ko Tzu-Yu , Li Chuan , Yang Jiann-Jou
Published: May 2014

Hearing loss is the most common sensory disorder, worldwide. In a recent study, we have identified a missense mutation, p.V174M, in the connexin 31 encoded by the GJB3 gene, in a patient with nonsyndromic hearing loss. However, the functional change in the CX31V174M mutant remains unknown. This study compared the intracellular distribution and assembly of the mutant CX31V174M with that of the wild-type (WT) CX31 in HeLa cells, and it examined the effect that the mutant protein had on those cells. A fluorescent localization assay of WT CX31 showed the typical punctuate pattern of a gap junction channel between the neighboring expression cells. Conversely, the p.V174M missense mutation resulted in the accumulation of the mutant protein in the lysosomes rather than in the cytoplasmic membrane. Moreover, dye transfer experiments have also demonstrated that the CX31V174M mutant did not form functional gap junction channels, probably due to the incorrect assembly or the altered properties of the CX31 channels. In addition, we found that CX31V174M-transfection can cause cell death by MTT assay. CX31V174M co-expressed with either CX31WT or CX26WT studies, suggested the impairment of the ability of CX26WT proteins to intracellular trafficking and targeting to the plasma membrane, but did not influence the trafficking of CX31WT. Based on these findings, we suggest that the CX31V174M mutant may have an effect on the formation and function of the gap junction, and CX31V174M has a trans-dominant negative effect on the function of wild types CX26. These results provide a novel molecular explanation for the role that GJB3 plays in hearing loss.
Consistency and universality in odd and even dimensional space–time QFT perturbative calculations
Authors: Battistel O. A. , Dallabona G.
Published: May 2014

The questions related to the consistent interpretation of QFT perturbative amplitudes are considered in light of a novel procedure, alternative to the traditional ones based on regularization prescriptions. A detailed discussion about the aspects associated to the space–time dimension is performed. For this purpose, it is considered a simple model having a fermionic vector current, coupled to a vector field, as well as a fermionic scalar current, coupled to a scalar field, both of them composed by different species of massive fermions. The referred currents are related in a precise way, which is reflected in the Ward identities for the perturbative physical amplitudes. The double vector two-point fermionic function, related to the vacuum polarization tensor of QED, as well as the amplitudes related to such quantity through relations among Green functions are explicit evaluated in space–time dimensions d = 2, 3, 4, 5 and 6. In the adopted procedure the perturbative amplitudes are not modified in intermediary steps of the calculations, as occurs in regularization procedures. Divergent Feynman integrals are not really solved. They appear only in standard objects, conveniently defined, where no physical parameter is present. Only very general properties for such quantities are assumed. For the finite parts, a set of functions is introduced which allows universal forms for the results. We show that scale independent, ambiguity free amplitudes are automatically obtained in a regularization independent way. As a consequence, interesting and, in certain way, surprising aspects are revealed in a clear and transparent way when the Ward identities and low-energy limits are verified for the simple amplitudes considered in the presently reported investigation. The obtained results suggest that the procedure can be considered as an advantageous tool to handle with the problem of divergences in perturbative solutions of QFT's, relative to the traditional regularization techniques, since the obtained results are so consistent as desirable and there are no limitations of applicability. In particular, the method can be applied in odd and even space–time dimensions having extra dimensions, which is not possible within the context of traditional regularization.
Non-weight bearing-induced muscle weakness: the role of myosin quantity and quality in MHC type II fibers
Authors: Kim Jong-Hee , Thompson LaDora V.
Published: May 2014

We tested the hypothesis that non-weight bearing-induced muscle weakness (i.e., specific force) results from decreases in myosin protein quantity (i.e., myosin content per half-sarcomere and the ratio of myosin to actin) and quality (i.e., force per half-sarcomere and population of myosin heads in the strong-binding state during muscle contraction) in single myosin heavy chain (MHC) type II fibers. Fisher-344 rats were assigned to weight-bearing control (Con) or non-weight bearing (NWB). The NWB rats were hindlimb unloaded for 2 wk. Diameter, force, and MHC content were determined in permeabilized single fibers from the semimembranosus muscle. MHC isoform and the ratio of MHC to actin in each fiber were determined by gel electrophoresis and silver staining techniques. The structural distribution of myosin from spin-labeled fiber bundles during maximal isometric contraction was evaluated using electron paramagnetic resonance spectroscopy. Specific force (peak force per cross-sectional area) in MHC type IIB and IIXB fibers from NWB was significantly reduced by 38% and 18%, respectively. MHC content per half-sarcomere was significantly reduced by 21%. Two weeks of hindlimb unloading resulted in a reduced force per half-sarcomere of 52% and fraction of myosin strong-binding during contraction of 34%. The results suggest that reduced myosin and actin content (quantity) and myosin quality concomitantly contribute to non-weight bearing-related muscle weakness.
Vagal afferents are not necessary for the satiety effect of the gut lipid messenger oleoylethanolamide
Authors: Karimian Azari Elnaz , Ramachandran Deepti , Weibel Sandra , Arnold Myrtha , Romano Adele , Gaetani Silvana , Langhans Wolfgang , Mansouri Abdelhak
Published: May 2014

The endogenous lipid messenger oleoylethanolamide (OEA) inhibits eating and modulates fat metabolism supposedly through the activation of peroxisome proliferator-activated receptor-α (PPARα) and vagal sensory fibers. We tested in adult male rats whether OEA stimulates fatty acid oxidation (FAO) and ketogenesis and whether it increases plasma levels of the satiating gut peptides glucagon-like peptide-1 (GLP-1) and peptide YY (PYY). We also explored whether OEA still inhibits eating after subdiaphragmatic vagal deafferentation (SDA). We found that intraperitoneally injected OEA (10 mg/kg body wt) reduced ( P < 0.05) food intake mainly by increasing meal latency and that this effect was stronger in rats fed a 60% high-fat diet (HFD) than in chow-fed rats. OEA increased ( P < 0.05) postprandial plasma nonesterified fatty acids and β-hydroxybutyrate (BHB) in the hepatic portal vein (HPV) and vena cava (VC) 30 min after injection, which was more pronounced in HFD- than in chow-fed rats. OEA also increased the protein expression of the key ketogenetic enzyme, mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase, in the jejunum of HFD-fed rats, but not in the liver or duodenum of either diet group. Furthermore, OEA decreased GLP-1 and PYY concentrations ( P < 0.05) in the HPV and VC 30 min after administration. Finally, OEA reduced food intake in SDA and sham-operated rats similarly. Our findings indicate that neither intact abdominal vagal afferents nor prandial increases in GLP-1 or PYY are necessary for the satiety effect of OEA. The enhanced FAO and ketogenesis raise the possibility of an involvement of intestine-derived BHB in OEA's satiety effect under certain conditions.
A role of nesfatin-1/NucB2 in dehydration-induced anorexia
Authors: Yoshimura Mitsuhiro , Matsuura Takanori , Ohkubo Junichi , Maruyama Takashi , Ishikura Toru , Hashimoto Hirofumi , Kakuma Tetsuya , Mori Masatomo , Ueta Yoichi
Published: May 2014

Nesfatin-1/NucB2, an anorexigenic molecule, is expressed mainly in the hypothalamus, particularly in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN). Nesfatin-1/NucB2 is also expressed in the subfornical organ (SFO). Because the SON and PVN are involved in body fluid regulation, nesfatin-1/NucB2 may be involved in dehydration-induced anorexia. To clarify the effects of endogenous nesfatin-1/NucB2, we studied changes in nesfatin-1/NucB2 mRNA levels in the SFO, SON, and PVN in adult male Wistar rats after exposure to osmotic stimuli by using in situ hybridization histochemistry. Significant increases in nesfatin-1/NucB2 mRNA levels, ∼2- to 3-fold compared with control, were observed in the SFO, SON, and PVN following water deprivation for 48 h, consumption of 2% NaCl hypertonic saline in drinking water for 5 days, and polyethylene glycol-induced hypovolemia. In addition, nesfatin-1/NucB2 expression was increased in response to water deprivation in a time-dependent manner. These changes in nesfatin-1/NucB2 mRNA expression were positively correlated with plasma sodium concentration, plasma osmolality, and total protein levels in all of the examined nuclei. Immunohistochemistry for nesfatin-1/NucB2 revealed that nesfatin-1/NucB2 protein levels were also increased after 48 h of dehydration and attenuated by 24 h of rehydration. Moreover, intracerebroventricular administration of nesfatin-1/NucB2-neutralizing antibody after 48 h of water deprivation resulted in a significant increase in food intake compared with administration of vehicle alone. These results suggested that nesfatin-1/NucB2 is a crucial peptide in dehydration-induced anorexia.