bluuebunny/crossref_metadata_2025 huggingface dataset:
Clark's In the Deep Midwinter and Hansen's Atticus: Examples of a Two-fold Literature of Life
Authors: Deavel Catherine Jack , Deavel David Paul
Published: July 2008

John Paul II's “Letter to Artists” identified two ways artists, particularly literary artists, can help reveal the nature of man to himself by showing both 1) the threat to human dignity from humans themselves, as well as 2) the possibility of transcendence and redemption as achievement and divine gift breaking into this life. We offer close readings of two contemporary novels as examples. The first way is illustrated by Robert Clark's In the Deep Midwinter, a novel at whose centre is an illegal abortion in the 1950's. We argue that the novel's portrait of suffering and abiding loss effectively shows the devastating effects of moral evil. The characters are conflicted in their desires and chosen actions, and they defend different positions; however, the plot in particular underscores the harm humans can inflict on themselves and others. The second way is illustrated using Hansen's Atticus. We argue that the character of Atticus serves both as an example of a virtuous Christian everyman and as an allegorical representation of God the Father. Redemption becomes possible for the dissolute son Scott when he turns to Atticus, his loving father, for forgiveness.
In vitro expansion of DNA triplet repeats with bulge binders and different DNA polymerases
Authors: Ouyang Di , Yi Long , Liu Liangliang , Mu Hong‐Tao , Xi Zhen
Published: July 2008

The expansion of DNA repeat sequences is associated with many genetic diseases in humans. Simple bulge DNA structures have been implicated as intermediates in DNA slippage within the DNA repeat regions. To probe the possible role of bulged structures in DNA slippage, we designed and synthesized a pair of simple chiral spirocyclic compounds [Xi Z, Ouyang D & Mu HT (2006) Bioorg Med Chem Lett16, 1180–1184], DDI‐1A and DDI‐1B, which mimic the molecular architecture of the enediyne antitumor antibiotic neocarzinostatin chromophore. Both compounds strongly stimulated slippage in various DNA repeats in vitro. Enhanced slippage synthesis was found to be synchronous for primer and template. CD spectra and UV thermal stability studies supported the idea that DDI‐1A and DDI‐1B exhibited selective binding to the DNA bulge and induced a significant conformational change in bulge DNA. The proposed mechanism for the observed in vitro expansion of long DNA is discussed.
Global shape and pH stability of ovorubin, an oligomeric protein from the eggs of Pomacea canaliculata
Authors: Dreon Marcos S. , Ituarte Santiago , Ceolín Marcelo , Heras Horacio
Published: July 2008

Ovorubin, a 300‐kDa thermostable oligomer, is the major egg protein from the perivitellin fluid that surrounds the embryos of the apple snail Pomacea canaliculata. It plays essential roles in embryo development, including transport and protection of carotenoids, protease inhibition, photoprotection, storage, and nourishment. Here, we report ovorubin dimensions and global shape, and test the role of electrostatic interactions in conformational stability by analyzing the effects of pH, using small‐angle X‐ray scattering (SAXS), transmission electron microscopy, CD, and fluorescence and absorption spectroscopy. Analysis of SAXS data shows that ovorubin is an anisometric particle with a major axis of 130 Å and a minor one varying between 63 and 76 Å. The particle shape was not significantly affected by the absence of the cofactor astaxanthin. The 3D model presented here is the first for an invertebrate egg carotenoprotein. The quaternary structure is stable over a wide pH range (4.5–12.0). At a pH between 2.0 and 4.0, a reduction in the gyration radius and a loss of tertiary structure are observed, although astaxanthin binding is not affected and only minor alterations in secondary structure are observed. In vitro pepsin digestion indicates that ovorubin is resistant to this protease action. The high stability over a considerable pH range and against pepsin, together with the capacity to bear temperatures > 95 °C, reinforces the idea that ovorubin is tailored to withstand a wide variety of conditions in order to play its key role in embryo protection during development.
Australasian emergency physicians: A Learning and educational needs analysis. Part Five: Barriers to CPD experienced by FACEM, and attitudes to the ACEM MOPS programme
Authors: Dent Andrew W , Weiland Tracey J , Paltridge Debbie
Published: July 2008

Object:  To determine the barriers to continuous professional development (CPD) perceived by Fellows of the Australasian College for Emergency Medicine (FACEM), their opinions on the maintenance of professional standards programme (MOPS), and other aspects of education.

Method:  Mailed survey offering 12 factors that could prevent them from participating in CPD. FACEM were asked to rate the items on Likert from 1, no barrier, through to 5, insurmountable barrier. Qualitative comments on barriers, MOPS and other education were grouped.

Results:  Nearly 47% of FACEM nominated lack of time with too many other priorities as significant of insurmountable barriers to CPD whilst 41% cited their ED as being too busy to leave and 35% had family commitments preventing CPD. While commitments, resistance from colleagues or employers (7%), lack of personal motivation or interest, (7.5%) lack of conference leave (8%) or financial restraint (14.5%) were lesser barriers for FACEM. Scheduling at inconvenient times (25%) and geographical factors (19%) were also perceived as significant barriers. Analysis of qualitative responses highlighted staff shortages, family reasons, personal fatigue, lack of interest in available CPD, cost and limited nonclinical time as being important barriers for some FACEM. Less than 50% of FACEM agreed that current allocation of MOPS points was appropriate or that the website spot diagnoses and scenarios kept their knowledge current. Nearly 28% did not find online MOPS easy to comprehend.

Conclusion:  These findings may assist in facilitating increased participation in CPD by FACEM.
Review article: Leaving the emergency department without being seen
Authors: Kennedy Marcus , MacBean Catherine E , Brand Caroline , Sundararajan Vijaya , McD Taylor David
Published: July 2008

Patients who leave the ED without being seen (LWBS) are unlikely to be satisfied with the quality of the service provided and might be at risk from conditions that have not been assessed or treated. We therefore examined the available research literature to inform the following questions: (i) In patients who attend for ED care, what factors are associated with the decision to LWBS? (ii) In patients who attend for ED care, are there adverse health outcomes associated with the decision to LWBS? (iii) Which interventions have been used to try to reduce the number of patients who attend for ED care and LWBS? From the available literature, there was insufficient evidence to draw firm conclusions; however, the literature does suggest that patients who LWBS have conditions of lower urgency and lower acuity, are more likely to be male and younger, and are likely to identify prolonged waiting times as a central concern. LWBS patients generally have very low rates of subsequent admission, and reports of serious adverse events are rare. Many LWBS patients go on to seek alternative medical attention, and they might have higher rates of ongoing symptoms at follow‐up. Further research is recommended to include comprehensive cohort or well‐designed case–control studies. These studies should assess a wide range of related factors, including patient, hospital and other relevant factors. They should compare outcomes for groups of LWBS patients with those who wait and should include cross‐sectoral data mapping to truly detect re‐attendance and admission rates.
Re‐presentation and suicide rates in emergency department patients who self‐harm
Authors: Howson Mark A , Yates Kim M , Hatcher Simon
Published: July 2008

Objective:  To investigate adult presentations to North Shore Hospital, Auckland, New Zealand, following deliberate self‐harm, determine re‐presentation and suicide rates and investigate the characteristics of those patients who re‐present.

Methods:  Retrospective review of a cohort of adult patients presenting to North Shore Hospital Emergency Care Centre in Auckland, New Zealand, with deliberate self‐harm between 1 January 2001 and 31 August 2002, using data from the National Minimum Data Set and Hospital Discharge Database. The re‐presentation rate was calculated as the percentage of patients presenting a second time within 1 year of their index presentation. The suicide rate was calculated as the percentage of patients with a coroner's verdict of suicide within 1 year of their index presentation of deliberate self‐harm.

Results:  There were 1055 presentations by 754 people during the presentation period. Of these 754 people, 136 presented for a second time within a year of the index case (136/754 [18.0%, 95% CI 15.5–21.0]). Eight of the seven hundred and fifty‐four people went on to commit suicide within 1 year (8/754 [1.1%, 95% CI 0.5–2.1]). Self‐poisoning was the most common method of self‐harm. Minor tranquillizers were used more commonly in the group who re‐presented.

Conclusion:  Re‐presentation and suicide rates in our study were similar to previously published rates. Further research is needed to identify strategies to reduce re‐presentations and excess mortality in all people who deliberately self‐harm.
Introduction of emergency medicine in China
Authors: Hou Xiang‐Yu , FitzGerald Gerard
Published: July 2008

Over the past 20 years, emergency medicine (EM) in China has been through a period of rapid development. This included the formal establishment of professional association of EM in 1986 and the establishment of ED in all county‐level hospitals across the country in the late 1990s. In line with the rapid economic development of China, ED have been equipped with appropriate ‘hardware’ equipment, but the ‘software’ part of the ED system remains underdeveloped. Doctors do not usually work exclusively in ED, but on a rotational basis, while also working as specialists in their own departments, such as medicine and surgery. EM in China remains underdeveloped, at least partly, for two main reasons: the current financial status of the health‐care system and lack of sufficient numbers of qualified EM specialists. Chinese education and training systems are now beginning to produce high‐quality emergency specialists, although there is not yet consistency across all courses. In Australia, the specialty of EM is well developed and, as such, this country is well placed to contribute to the development of ED in China.
Australasian emergency physicians: A learning and educational needs analysis. Part six: Differences in confidence, exposure to learning and expressed learning needs by subgroups of Australasian emergency physicians
Authors: Dent Andrew W , Paltridge Debbie , Weiland Tracey J
Published: July 2008

Objective:  To determine differences in the confidence and learning needs between subgroups of Fellows of the Australasian College for Emergency Medicine (FACEM), according to sex, type of hospital of principal employment, region of practice in Australasia, year of obtaining the fellowship and percentage of hours worked clinically.

Method:  Mailed survey of all FACEM with multiple Likert scale options.

Results:  Male emergency physicians (EP) more often expressed confidence in 14 of 58 tasks, but female EP more often expressed a strong desire for continuing professional development (CPD) in 8. EP working in private hospitals were more likely to find lack of conference in their contracts a barrier to participation in CPD (28% [CI 14.1–47.8]vs 7.3% [CI 5.2–10.1]; P = 0.003). EP obtaining their fellowship before 1990 were more confident in 13 of 25 non‐clinical tasks than others but had similar confidence in clinical skills. Although EP who spent 30% or less on clinical work expressed confidence more frequently in a range of non‐clinical skills, they were less confident for paediatric emergencies, central lines, orthopaedics and tube thoracostomy. There were few variations between Australian states and territories and New Zealand in confidence of EP. EP in regional and rural centres nominated geographical factors (31.3% [CI 21.2–43.4]vs 17.2% [CI 13.9–21.3]; P = 0.012), other colleagues being at the same conference (46.9% [CI 35.2–58.9]vs 28.3% [CI 24.2–32.9]; P = 0.005) and inability to use their conference leave (48.4% [CI 36.6–60.4]vs 33.1% [CI 28.7–37.8]; P = 0.024) as significant barriers more frequently than their city colleagues.

Conclusion:  These findings might assist planning CPD for EP.
Accuracy of current clinical diagnosis in recreational drug‐related attendance to the emergency department
Authors: West Emma , Cameron Peter , O'Reilly Gerard , Drummer Olaf H , Bystrzycki Adam
Published: July 2008

Objective:  To determine the accuracy of current clinical diagnosis in recreational drug‐related attendances to emergency by blood analysis.

Methods:  A prospective convenience sample of 103 patients who attended hospital with suspected recreational drug‐related presentations was collected. Doctors' clinical impression of drugs responsible for presentation was compared with a detailed forensic blood analysis for recreational drugs.

Results:  Among 103 samples, 80 (78%, 95% confidence intervals [CI] 70–86%) were found to have correct clinical suspicion of the recreational drug responsible for clinical presentation confirmed by laboratory analysis. Clinical diagnosis was most accurate for gamma‐hydroxy butyrate (GHB) (sensitivity 97%, specificity 91%) and less accurate for amphetamines (sensitivity 61%, specificity 79%), alcohol (sensitivity 42%, specificity 84%) and opiates (sensitivity 46%, specificity 100%). Multiple drug ingestion was found in 70% (95% CI 61–79%) of samples. Sensitivity and specificity of clinical impression for prediction of multiple drug ingestion presence is 75% (95% CI 66–83%) and 85% (95% CI 78–92%), respectively.

Conclusion:  Clinical diagnosis in recreational drug‐related attendances to the ED was correct in most cases. Drugs, such as GHB, were the most accurately diagnosed. Inaccuracy in recognizing other drugs, like amphetamines, opiates and alcohol, occurs where a coingestant produces a more profound clinical picture. Multiple drug ingestion is a common scenario in recreational drug presentations to emergency.
North Pacific right whale (Eubalaena japonica) seasonal and diel calling patterns from long‐term acoustic recordings in the southeastern Bering Sea, 2000–2006
Authors: Munger Lisa M. , Wiggins Sean M. , Moore Sue E. , Hildebrand John A.
Published: July 2008

Abstract

We assessed North Pacific right whale (Eubalaena japonica) seasonal and daily calling patterns in the southeastern Bering Sea (SEBS) using long‐term hydrophone recordings from October 2000 through January 2006. We detected right whale calls on the SEBS middle shelf (<100 m depth) as early as May, intermittently throughout summer and fall, and as late as December. Calls also were detected on one day in June 2005 on the SEBS slope (>1,000 m), but were not detected near Kodiak Island from April to August 2003. In months with calls, detections occurred on more days in July–October (≥6 d/mo), than from May to June or November to December (≤3 d/mo). Calls were clustered in time and were usually detected on 1–3 consecutive days with a median interval of 6.5 d for calls >1 d apart. Hourly calling rates were significantly higher at night than during the day. These data indicate that right whales occur in the SEBS later in the year than previously known, intermittently pass through the middle‐shelf study region, and usually remain there no longer than a few days. Right whale habitat use in the SEBS may intensify in mid‐summer through early fall based on higher monthly and daily call detection rates.
The English Student Movement: An Evaluation of the Literature
Authors: Hanna Esmee
Published: July 2008

This article explores the state of the field of student movement research. I suggest there could be seen to be stagnation within the field of investigation, and resultant under‐researching of some countries student movements, and I will make specific reference to the student movement of England in the late 1960s/early 1970s as a case in point of this. I argue that there has been unsatisfactory sole‐causalities, such as issues of youth, issues seen as ‘triggers’, and political factors, attributed to the English student movement, and that this fails both to understand fully the significance and individuality of the English student movement, but also assumes a fit with New social movement (NSM) theory. I argue that we cannot automatically equate NSM's and student movements without thorough empirical research, and that we need to look to a synthesis of social movement theories in order to understand fully student movements.
From Iraq to Katrina and Back: Bureaucratic Planning as Strategic Failure, Fiction, and Fantasy
Authors: Bissell William Cunningham
Published: July 2008

Bureaucratic planning is a complex sociocultural process rife with vested interests, contradictory aims, and internal conflicts. In the wake of Hurricane Katrina and the US occupation of Iraq, contemporary media discourses on planning have downplayed this complexity and diverted attention away from a more nuanced understanding of planning as a sociological process. In this article, I show how policy and press debates regarding bureaucratic planning in Iraq and New Orleans are rooted in technocratic and rationalist perspectives, constituting a kind of modernist faith. These contemporary discourses are often one‐dimensional, failing to consider the conflicts, contradictions, and counter‐intuitive outcomes that are inherent in planning in complex situations, including urban reconstruction, military occupation, and disaster response. I show how flawed approaches in the present are deeply linked to the past, connecting Iraq and Katrina to a detailed ethnographic case study on failed urban planning in colonial Zanzibar. Even though the three cases of Iraq, Katrina, and Zanzibar differ across time, context, and conditions, they reveal that bureaucratic planning is best understood as a multidimensional strategic practice that is intrinsically shaped by sociocultural forms. Plans may ostensibly fail in one sense and yet still produce quite enduring effects. They may remain ‘fictions’ at one level but still wield real impact. And even if they are informed by fantasy, bearing little utility from a functional perspective, plans still serve as performative and persuasive acts, informed by political ends.
The Ethical Dilemma of Ethical Committees
Authors: Derbyshire Stuart
Published: July 2008

The Nuremberg code, a response to the 1946 Nuremberg Medical Trials, was the first attempt to formally state ethical requirements for medical research. The Code was generally ignored as a response to the peculiarly barbaric Nazi atrocities and an unnecessary fetter on normal research. A series of research scandals, however, led to more successful attempts at regulating medical research and to the introduction of various ethical committees during the 1970s. Since then, ethical committees have expanded their remit to regulate social as well as medical research and operate according to precautionary standards that far exceed what is necessary to protect public safety. Ethical committees block investigations of medical practice even when the intent is to benefit patients directly and they prevent social research entailing even far‐fetched possibilities of ‘stress’. Although purportedly designed to protect patients and civil liberties, modern ethical regulation damages the doctor–patient relationship, undermines professional responsibility, and encourages negative scientific practice.
Colossal Ionic Conductivity at Interfaces of Epitaxial ZrO
2
:Y
2
O
3
/SrTiO
3
Heterostructures
Authors: Garcia-Barriocanal J. , Rivera-Calzada A. , Varela M. , Sefrioui Z. , Iborra E. , Leon C. , Pennycook S. J. , Santamaria J.
Published: July 2008

The search for electrolyte materials with high oxygen conductivities is a key step toward reducing the operation temperature of fuel cells, which is currently above 700°C. We report a high lateral ionic conductivity, showing up to eight orders of magnitude enhancement near room temperature, in yttria-stabilized zirconia (YSZ)/strontium titanate epitaxial heterostructures. The enhancement of the conductivity is observed, along with a YSZ layer thickness–independent conductance, showing that it is an interface process. We propose that the atomic reconstruction at the interface between highly dissimilar structures (such as fluorite and perovskite) provides both a large number of carriers and a high-mobility plane, yielding colossal values of the ionic conductivity.
The Cell and Molecular Basis of Mechanical, Cold, and Inflammatory Pain
Authors: Abrahamsen Bjarke , Zhao Jing , Asante Curtis O. , Cendan Cruz Miguel , Marsh Steve , Martinez-Barbera Juan Pedro , Nassar Mohammed A. , Dickenson Anthony H. , Wood John N.
Published: July 2008

Peripheral pain pathways are activated by a range of stimuli. We used diphtheria toxin to kill all mouse postmitotic sensory neurons expressing the sodium channel Na
v
1.8. Mice showed normal motor activity and low-threshold mechanical and acute noxious heat responses but did not respond to noxious mechanical pressure or cold. They also showed a loss of enhanced pain responses and spontaneous pain behavior upon treatment with inflammatory insults. In contrast, nerve injury led to heightened pain sensitivity to thermal and mechanical stimuli indistinguishable from that seen with normal littermates. Pain behavior correlates well with central input from sensory neurons measured electrophysiologically in vivo. These data demonstrate that Na
v
1.8-expressing neurons are essential for mechanical, cold, and inflammatory pain but not for neuropathic pain or heat sensing.
Pyogenic Bacterial Infections in Humans with MyD88 Deficiency
Authors: von Bernuth Horst , Picard Capucine , Jin Zhongbo , Pankla Rungnapa , Xiao Hui , Ku Cheng-Lung , Chrabieh Maya , Mustapha Imen Ben , Ghandil Pegah , Camcioglu Yildiz , Vasconcelos Júlia , Sirvent Nicolas , Guedes Margarida , Vitor Artur Bonito , Herrero-Mata María José , Aróstegui Juan Ignacio , Rodrigo Carlos , Alsina Laia , Ruiz-Ortiz Estibaliz , Juan Manel , Fortuny Claudia , Yagüe Jordi , Antón Jordi , Pascal Mariona , Chang Huey-Hsuan , Janniere Lucile , Rose Yoann , Garty Ben-Zion , Chapel Helen , Issekutz Andrew , Maródi László , Rodriguez-Gallego Carlos , Banchereau Jacques , Abel Laurent , Li Xiaoxia , Chaussabel Damien , Puel Anne , Casanova Jean-Laurent
Published: July 2008

MyD88 is a key downstream adapter for most Toll-like receptors (TLRs) and interleukin-1 receptors (IL-1Rs). MyD88 deficiency in mice leads to susceptibility to a broad range of pathogens in experimental settings of infection. We describe a distinct situation in a natural setting of human infection. Nine children with autosomal recessive MyD88 deficiency suffered from life-threatening, often recurrent pyogenic bacterial infections, including invasive pneumococcal disease. However, these patients were otherwise healthy, with normal resistance to other microbes. Their clinical status improved with age, but not due to any cellular leakiness in MyD88 deficiency. The MyD88-dependent TLRs and IL-1Rs are therefore essential for protective immunity to a small number of pyogenic bacteria, but redundant for host defense to most natural infections.
Evidence of Global Chlorophyll d
Authors: Kashiyama Y. , Miyashita H. , Ohkubo S. , Ogawa N. O. , Chikaraishi Y. , Takano Y. , Suga H. , Toyofuku T. , Nomaki H. , Kitazato H. , Nagata T. , Ohkouchi N.
Published: July 2008

Although analyses of chlorophyll d (Chl d)‐dominated oxygenic photosystems have been conducted since their discovery 12 years ago, Chl d distribution in the environment and quantitative importance for aquatic photosynthesis remain to be investigated. We analyzed the pigment compositions of surface sediments and detected Chl d and its derivatives from diverse aquatic environments. Our data show that the viable habitat for Chl d‐producing phototrophs extends across salinities of 0 to 50 practical salinity units and temperatures of 1� to 40�C, suggesting that Chl d production can be ubiquitously observed in aquatic environments that receive near-infrared light. The relative abundances of Chl d derivatives over that of Chl a derivatives in the studied samples are up to 4%, further suggesting that Chl d‐based photosynthesis plays a quantitatively important role in the aquatic photosynthesis.
The Global Stoichiometry of Litter Nitrogen Mineralization
Authors: Manzoni Stefano , Jackson Robert B. , Trofymow John A. , Porporato Amilcare
Published: July 2008

Plant residue decomposition and the nutrient release to the soil play a major role in global carbon and nutrient cycling. Although decomposition rates vary strongly with climate, nitrogen immobilization into litter and its release in mineral forms are mainly controlled by the initial chemical composition of the residues. We used a data set of ∼2800 observations to show that these global nitrogen-release patterns can be explained by fundamental stoichiometric relationships of decomposer activity. We show how litter quality controls the transition from nitrogen accumulation into the litter to release and alters decomposers' respiration patterns. Our results suggest that decomposers lower their carbon-use efficiency to exploit residues with low initial nitrogen concentration, a strategy used broadly by bacteria and consumers across trophic levels.
Structures of Neutral Au
7
, Au
19
, and Au
20
Clusters in the Gas Phase
Authors: Gruene Philipp , Rayner David M. , Redlich Britta , van der Meer Alexander F. G. , Lyon Jonathan T. , Meijer Gerard , Fielicke André
Published: July 2008

The catalytic properties of gold nanoparticles are determined by their electronic and geometric structures. We revealed the geometries of several small neutral gold clusters in the gas phase by using vibrational spectroscopy between 47 and 220 wavenumbers. A two-dimensional structure for neutral Au
7
and a pyramidal structure for neutral Au
20
can be unambiguously assigned. The reduction of the symmetry when a corner atom is cut from the tetrahedral Au
20
cluster is directly reflected in the vibrational spectrum of Au
19
.
Distinctive Features of Surface-Anchored Proteins of
Streptococcus agalactiae
Strains from Zimbabwe Revealed by PCR and Dot Blotting
Authors: Mavenyengwa Rooyen T. , Maeland Johan A. , Moyo Sylvester R.
Published: July 2008

The distribution of capsular polysaccharide (CPS) types and subtypes (serovariants) among 121 group B streptococcus (GBS) strains from Zimbabwe was examined. PCR was used for the detection of both CPS types and the surface-anchored and strain-variable proteins Cα, Cβ, Alp1, Alp2, Alp3, R4/Rib, and Alp4. The R3 protein was detected by an antibody-based method using monoclonal anti-R3 antibody in dot blotting. The CPS types detected, Ia (15.7% of strains), Ib (11.6%), II (8.3%), III (38.8%), V (24.0%), and nontypeable (1.7%), were essentially as expected on the basis of data from Western countries. The type V strains showed distinctive features with respect to protein markers in that Alp3 was detected in only 6.9% of the isolates while R3 occurred in 75.9% and R4/Rib occurred in 37.9% of the isolates. R3 occurred nearly always in combination with one of the alpha-like (Alp) proteins, and it was the third most common of the proteins studied. These results show that type V GBS strains from Zimbabwe differed from type V strains from other geographical areas and also emphasize the importance of the R3 protein in GBS serotyping and its potential importance in the immunobiology of GBS, including a potential role in a future GBS vaccine.
Empyema Necessitatis Due to Methicillin-Resistant
Staphylococcus aureus
: Case Report and Review of the Literature
Authors: Mizell Kelly N. , Patterson Kimberley V. , Carter J. Elliot
Published: July 2008

Empyema necessitatis is a rare complication of empyema in which the pleural infection spreads outside of the pleural space to involve the soft tissues of the chest wall. Most cases of empyema necessitatis are related to
Mycobacterium tuberculosis
and, less commonly, to
Actinomyces
spp. and
Streptococcus
spp.
Staphylococcus aureus
has rarely been reported as the causative agent of empyema necessitatis, with the majority of
S. aureus
isolates being methicillin sensitive. Only two cases of empyema necessitatis due to methicillin-resistant
S. aureus
(MRSA) have been reported in the medical literature. We report the case of a 59-year-old Caucasian male who presented to our institution with complaints of pain in and swelling of his left upper chest of 2-months duration. A computed tomography scan of the chest showed an 8.1- by 6.5-cm lesion which extended from the left upper lobe of the lung into the extrathoracic soft tissues beneath the left upper pectoralis muscle. A wedge resection of the left upper lung lobe revealed lung tissue with an organized pneumonia-like pattern associated with marked acute pleuritis. Blood and urine cultures and cultures of the left chest soft tissue mass grew MRSA. The patient was successfully treated with vancomycin followed by a 10-day outpatient course of ciprofloxacin and trimethoprim-sulfamethoxazole. This case represents an extremely rare manifestation of an increasingly dangerous bacterial pathogen.
Prevalence of Human KI and WU Polyomaviruses in Children with Acute Respiratory Tract Infection in China
Authors: Yuan Xin-hui , Jin Yu , Xie Zhi-ping , Gao Han-chun , Xu Zi-qian , Zheng Li-shu , Zhang Rong-fang , Song Jing-rong , Hou Yun-de , Duan Zhao-jun
Published: July 2008

The KI and WU polyomaviruses were found in 11 (2.7%) and 17 (4.2%) of 406 nasopharyngeal aspirates, respectively, from children with acute respiratory tract infection (ARTI). The phylogenetic analysis indicates that they are all in the same cluster as the prototype strains. Our findings suggest that they are common in children with ARTI in China.
Mutational Evidence for a Structural Model of the Lassa Virus RNA Polymerase Domain and Identification of Two Residues, Gly1394 and Asp1395, That Are Critical for Transcription but Not Replication of the Genome
Authors: Hass Meike , Lelke Michaela , Busch Carola , Becker-Ziaja Beate , Günther Stephan
Published: July 2008

The RNA-dependent RNA polymerase (RdRp) of arenaviruses is an integral part of the L protein, a 200-kDa multifunctional and multidomain protein. In view of the paucity of structural data, we recently proposed a model for the RdRp domain of arenaviruses based on the folding of RdRps of plus-strand viruses (S. Vieth et al., Virology 318:153-168, 2004). In the present study, we have chosen a large-scale mutagenesis approach to gain insight into the structure and function of the Lassa virus RdRp domain. A total of 180 different mutants of the domain were generated by using a novel PCR-based mutagenesis technique and tested in the context of the Lassa virus replicon system. Nearly all residues, which were essential for function, clustered in the center of the three-dimensional model including the catalytic site, while residues that were less important for function mapped to the periphery of the model. The combined bioinformatics and mutagenesis data allowed deducing candidate residues for ligand interaction. Mutation of two adjacent residues in the putative palm-thumb subdomain junction, G1394 and D1395 (strain AV), led to a defect in mRNA synthesis but did not affect antigenomic RNA synthesis. In conclusion, the data provide circumstantial evidence for the existence of an RdRp domain between residues 1040 and 1540 of the Lassa virus L protein and the folding model of the domain. A functional element within the RdRp was identified, which is important for transcription but not replication of the genome.
The DNA Damage Sensors Ataxia-Telangiectasia Mutated Kinase and Checkpoint Kinase 2 Are Required for Hepatitis C Virus RNA Replication
Authors: Ariumi Yasuo , Kuroki Misao , Dansako Hiromichi , Abe Ken-Ichi , Ikeda Masanori , Wakita Takaji , Kato Nobuyuki
Published: July 2008

Cellular responses to DNA damage are crucial for maintaining genome integrity, virus infection, and preventing the development of cancer. Hepatitis C virus (HCV) infection and the expression of the HCV nonstructural protein NS3 and core protein have been proposed as factors involved in the induction of double-stranded DNA breaks and enhancement of the mutation frequency of cellular genes. Since DNA damage sensors, such as the ataxia-telangiectasia mutated kinase (ATM), ATM- and Rad3-related kinase (ATR), poly(ADP-ribose) polymerase 1 (PARP-1), and checkpoint kinase 2 (Chk2), play central roles in the response to genotoxic stress, we hypothesized that these sensors might affect HCV replication. To test this hypothesis, we examined the level of HCV RNA in HuH-7-derived cells stably expressing short hairpin RNA targeted to ATM, ATR, PARP-1, or Chk2. Consequently, we found that replication of both genome-length HCV RNA (HCV-O, genotype 1b) and the subgenomic replicon RNA were notably suppressed in ATM- or Chk2-knockdown cells. In addition, the RNA replication of HCV-JFH1 (genotype 2a) and the release of core protein into the culture supernatants were suppressed in these knockdown cells after inoculation of the cell culture-generated HCV. Consistent with these observations, ATM kinase inhibitor could suppress the HCV RNA replication. Furthermore, we observed that HCV NS3-NS4A interacted with ATM and that HCV NS5B interacted with both ATM and Chk2. Taken together, these results suggest that the ATM signaling pathway is critical for HCV RNA replication and may represent a novel target for the clinical treatment of patients with chronic hepatitis C.
The Kinetics of Hepatitis C Virus-Specific CD8 T-Cell Responses in the Blood Mirror Those in the Liver in Acute Hepatitis C Virus Infection
Authors: Shin Eui-Cheol , Capone Stefania , Cortese Riccardo , Colloca Stefano , Nicosia Alfredo , Folgori Antonella , Rehermann Barbara
Published: July 2008

Peripheral blood T-cell responses are used as biomarkers in hepatitis C virus (HCV) vaccine trials. However, it is not clear how T-cell responses in the blood correlate with those in the liver, the infection site. By studying serial liver and blood samples of five vaccinated and five mock-vaccinated control chimpanzees during acute HCV infection, we demonstrate a correlation between HCV-specific CD8 T-cell responses in the blood and molecular and functional markers of T-cell responses in the liver. Thus, HCV-specific CD8 T-cell responses in the blood are valid markers for intrahepatic T-cell activity.
Scrapie-Induced Defects in Learning and Memory of Transgenic Mice Expressing Anchorless Prion Protein Are Associated with Alterations in the Gamma Aminobutyric Acid-Ergic Pathway
Authors: Trifilo Matthew J. , Sanchez-Alavez Manuel , Solforosi Laura , Bernard-Trifilo Joie , Kunz Stefan , McGavern Dorian , Oldstone Michael B. A.
Published: July 2008

After infection with RML murine scrapie agent, transgenic (tg) mice expressing prion protein (PrP) without its glycophosphatidylinositol (GPI) membrane anchor (GPI
−/−
PrP tg mice) continue to make abundant amounts of the abnormally folded disease-associated PrPres but have a normal life span. In contrast, all age-, sex-, and genetically matched mice with a GPI-anchored PrP become moribund and die due to a chronic progressive neurodegenerative disease by 160 days after RML scrapie agent infection. We report here that infected GPI
−/−
PrP tg mice, although free from progressive neurodegenerative disease of the cerebellum and extrapyramidal and pyramidal systems, nevertheless suffer defects in learning and memory, long-term potentiation, and neuronal excitability. Such dysfunction increases over time and is associated with an increase in gamma aminobutyric acid (GABA) inhibition but not loss of excitatory glutamate/
N
-methyl-
d
-aspartic acid. Enhanced deposition of abnormally folded infectious PrP (PrPsc or PrPres) in the central nervous system (CNS) localizes with GABAA receptors. This occurs with minimal evidence of CNS spongiosis or apoptosis of neurons. The use of monoclonal antibodies reveals an association of PrPres with GABAA receptors. Thus, the clinical defects of learning and memory loss in vivo in GPI
−/−
PrP tg mice infected with scrapie agent may likely involve the GABAergic pathway.
Mutations in Human Immunodeficiency Virus Type 1 Nucleocapsid Protein Zinc Fingers Cause Premature Reverse Transcription
Authors: Thomas James A. , Bosche William J. , Shatzer Teresa L. , Johnson Donald G. , Gorelick Robert J.
Published: July 2008

Human immunodeficiency virus type 1 (HIV-1) requires that its genome be reverse transcribed into double-stranded DNA for productive infection of cells. This process requires not only reverse transcriptase but also the nucleocapsid protein (NC), which functions as a nucleic acid chaperone. Reverse transcription generally begins once the core of the virion enters the cytoplasm of a newly infected cell. However, some groups have reported the presence of low levels of viral DNA (vDNA) within particles prior to infection, the significance and function of which is controversial. We report here that several HIV-1 NC mutants, which we previously identified as being replication defective, contain abnormally high levels of intravirion DNA. These findings were further reinforced by the inability of these NC mutants to perform endogenous reverse transcription (ERT), in contrast to the readily measurable ERT activity in wild-type HIV-1. When either of the NC mutations is combined with a mutation that inactivates the viral protease, we observed a significant reduction in the amount of intravirion DNA. Interestingly, we also observed high levels of intravirion DNA in the context of wild-type NC when we delayed budding by means of a PTAP
(−)
(Pro-Thr-Ala-Pro) mutation. Premature reverse transcription is most probably occurring before these mutant virions bud from producer cells, but we fail to see any evidence that the NC mutations alter the timing of Pr55
Gag
processing. Critically, our results also suggest that the presence of intravirion vDNA could serve as a diagnostic for identifying replication-defective HIV-1.
Herpes Simplex Virus Downregulates Secretory Leukocyte Protease Inhibitor: a Novel Immune Evasion Mechanism
Authors: Fakioglu Esra , Wilson Sarah S. , Mesquita Pedro M. M. , Hazrati Ehsan , Cheshenko Natalia , Blaho John A. , Herold Betsy C.
Published: July 2008

Secretory leukocyte protease inhibitor (SLPI), an anti-inflammatory mediator of mucosal immunity, inhibits human immunodeficiency virus (HIV) and herpes simplex virus (HSV) in cell culture. Epidemiological studies demonstrate that higher concentrations of SLPI in mucosal secretions are associated with a reduced risk of HIV transmission. The current studies were designed to test the hypothesis that HSV triggers a loss of SLPI to evade innate immunity and that this response may contribute to the increased risk of HIV infection in the setting of HSV infection. Exposure of human cervical epithelial cells to HSV-1 or HSV-2, but not HIV or vesicular stomatitis virus, triggered a significant and sustained reduction in SLPI levels. The reduction persisted when cells were infected in the presence of acyclovir but not following infection with UV-inactivated virus, indicating that viral gene expression, but not replication, is required. Reverse transcriptase PCR studies demonstrated that the loss of SLPI is mediated by downregulation of gene expression. SLPI downregulation was associated with activation of NF-κB signaling pathways and upregulation of proinflammatory cytokines, consistent with the known inhibitor effects of SLPI on NF-κB pathways. The downregulation mapped to viral early-gene expression, as variants impaired in expression of the ICP4 or ICP0 immediate-early gene failed to downregulate SLPI or activate NF-κB. Together, these results identify a novel role for HSV immediate-early-gene expression in regulating mucosal immune responses.
Potent Antibody-Mediated Neutralization and Evolution of Antigenic Escape Variants of Simian Immunodeficiency Virus Strain SIVmac239 In Vivo
Authors: Sato Shuji , Yuste Eloisa , Lauer William A. , Chang Eun Hyuk , Morgan Jennifer S. , Bixby Jacqueline G. , Lifson Jeffrey D. , Desrosiers Ronald C. , Johnson Welkin E.
Published: July 2008

Here, we describe the evolution of antigenic escape variants in a rhesus macaque that developed unusually high neutralizing antibody titers to SIVmac239. By 42 weeks postinfection, 50% neutralization of SIVmac239 was achieved with plasma dilutions of 1:1,000. Testing of purified immunoglobulin confirmed that the neutralizing activity was antibody mediated. Despite the potency of the neutralizing antibody response, the animal displayed a typical viral load profile and progressed to terminal AIDS with a normal time course. Viral envelope sequences from week 16 and week 42 plasma contained an excess of nonsynonymous substitutions, predominantly in V1 and V4, including individual sites with ratios of nonsynonymous to synonymous substitution rates (dN/dS) highly suggestive of strong positive selection. Recombinant viruses encoding envelope sequences isolated from these time points remained resistant to neutralization by all longitudinal plasma samples, revealing the failure of the animal to mount secondary responses to the escaped variants. Substitutions at two sites with significantdN/dSvalues, one in V1 and one in V4, were independently sufficient to confer nearly complete resistance to neutralization. Substitutions at three additional sites, one in V4 and two in gp41, conferred moderate to high levels of resistance when tested individually. All the amino acid changes leading to escape resulted from single nucleotide substitutions. The observation that antigenic escape resulted from individual, single amino acid replacements at sites well separated in current structural models of Env indicates that the virus can utilize multiple independent pathways to rapidly achieve similar levels of resistance.
Reduced Protection from Simian Immunodeficiency Virus SIV
mac251
Infection Afforded by Memory CD8
+
T Cells Induced by Vaccination during CD4
+
T-Cell Deficiency
Authors: Vaccari Monica , Mattapallil Joseph , Song Kaimei , Tsai Wen-Po , Hryniewicz Anna , Venzon David , Zanetti Maurizio , Reimann Keith A. , Roederer Mario , Franchini Genoveffa
Published: July 2008

Adaptive CD4
+
and CD8
+
T-cell responses have been associated with control of human immunodeficiency virus/simian immunodeficiency virus (HIV/SIV) replication. Here, we have designed a study with Indian rhesus macaques to more directly assess the role of CD8 SIV-specific responses in control of viral replication. Macaques were immunized with a DNA prime-modified vaccinia virus Ankara (MVA)-SIV boost regimen under normal conditions or under conditions of antibody-induced CD4
+
T-cell deficiency. Depletion of CD4
+
cells was performed in the immunized macaques at the peak of SIV-specific CD4
+
T-cell responses following the DNA prime dose. A group of naïve macaques was also treated with the anti-CD4 depleting antibody as a control, and an additional group of macaques immunized under normal conditions was depleted of CD8
+
T cells prior to challenge exposure to SIV
mac251
. Analysis of the quality and quantity of vaccine-induced CD8
+
T cells demonstrated that SIV-specific CD8
+
T cells generated under conditions of CD4
+
T-cell deficiency expressed low levels of Bcl-2 and interleukin-2 (IL-2), and plasma virus levels increased over time. Depletion of CD8
+
T cells prior to challenge exposure abrogated vaccine-induced protection as previously shown. These data support the notion that adaptive CD4
+
T cells are critical for the generation of effective CD8
+
T-cell responses to SIV that, in turn, contribute to protection from AIDS. Importantly, they also suggest that long-term protection from disease will be afforded only by T-cell vaccines for HIV that provide a balanced induction of CD4
+
and CD8
+
T-cell responses and protect against early depletion of CD4
+
T cells postinfection.
Apolipoprotein C1 Association with Hepatitis C Virus
Authors: Meunier Jean-Christophe , Russell Rodney S. , Engle Ronald E. , Faulk Kristina N. , Purcell Robert H. , Emerson Suzanne U.
Published: July 2008

Accumulating evidence suggests that cellular lipoprotein components are involved in hepatitis C virus (HCV) morphogenesis, but the precise contribution of these components remains unclear. We investigated the involvement of apolipoprotein C1 (ApoC1) in HCV infection in the HCV pseudotyped particle system (HCVpp), in the recently developed cell culture infection model (HCVcc), and in authentic HCV isolated from viremic chimpanzees. Viral genomes associated with HCVcc or authentic HCV were efficiently immunoprecipitated by anti-ApoC1, demonstrating that ApoC1 was a normal component of HCV. The infectivities of HCVpp that had been mixed with ApoC1 and, more importantly, untreated HCVcc collected from lysates or media of infected Huh7.5 cells were directly neutralized by anti-ApoC1. Indeed, convalescent anti-HCV immunoglobulin G and anti-ApoC1 each neutralized over 75% of infectious HCVcc particles, indicating that many, if not all, infectious particles were recognized by both antibodies. Moreover, peptides corresponding to the C-terminal region of ApoC1 blocked infectivity of both HCVpp and HCVcc. Altogether, these results suggest that ApoC1 associates intracellularly via its C-terminal region with surface components of virions during viral morphogenesis and may play a major role in the replication cycle of HCV.
Caveolin-1-Dependent Infectious Entry of Human Papillomavirus Type 31 in Human Keratinocytes Proceeds to the Endosomal Pathway for pH-Dependent Uncoating
Authors: Smith Jessica L. , Campos Samuel K. , Wandinger-Ness Angela , Ozbun Michelle A.
Published: July 2008

High-risk human papillomaviruses (HPVs) are small nonenveloped DNA viruses with a strict tropism for squamous epithelium. The viruses are causative agents of cervical cancer and some head and neck cancers, but their differentiation-dependent life cycles have made them difficult to study in simple cell culture. Thus, many aspects of early HPV infection remain mysterious. We recently showed the high-risk HPV type 31 (HPV31) enters its natural host cell type via caveola-dependent endocytosis, a distinct mechanism from that of the closely related HPV16 (Smith et al., J. Virol. 81:9922-9931, 2007). Here, we determined the downstream trafficking events after caveolar entry of HPV31 into human keratinocytes. After initial plasma membrane binding, HPV31 associates with caveolin-1 and transiently localizes to the caveosome before trafficking to the early endosome and proceeding through the endosomal pathway. Caveosome-to-endosome transport was found to be Rab5 GTPase dependent. Although HPV31 capsids were observed in the lysosome, Rab7 GTPase was dispensable for HPV31 infection, suggesting that viral genomes escape from the endosomal pathway prior to Rab7-mediated capsid transport. Consistent with this, the acidic pH encountered by HPV31 within the early endosomal pathway induces a conformational change in the capsid resulting in increased DNase susceptibility of the viral genome, which likely aids in uncoating and/or endosomal escape. The entry and trafficking route of HPV31 into human keratinocytes represents a unique viral pathway by which the virions use caveolar entry to eventually access a low-pH site that appears to facilitate endosomal escape of genomes.