bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential Distribution of Human Mitochondrial DNA in Somatic Tissues and Hairs
Authors: Lee Hwan Young , Chung Ukhee , Park Myung Jin , Yoo Ji‐Eun , Han Gil‐Ro , Shin Kyoung‐Jin
Published: January 2006

To investigate mitochondrial DNA (mtDNA) distribution within tissues during life, we observed length heteroplasmy in a polycytosine tract of the mitochondrial HV2 region by size‐based separation of PCR products, using a mutagenic primer which was designed to avoid stutter production. Blood, brain, heart, liver, skeletal muscle and hair shaft samples were collected during autopsies of 25 individuals. Here, we demonstrate differences in the level of mtDNA length heteroplasmy both within and between individuals and tissues. We also show that mtDNA is distributed randomly in varying proportions in various somatic tissues during growth, resulting in an imbalance in the composition of mtDNA pools among tissues. This mtDNA distribution appears not to be strictly random, and can be explained by the random somatic segregation of nucleoids. On the other hand, significant qualitative/quantitative mtDNA peak pattern variations in hair shafts are thought to be a result of the different developmental origins of hairs. Each hair shaft may have a restricted or clonal set of mtDNA molecules derived from a discrete group of stem cells.
Transfusion‐associated Chagas disease (American trypanosomiasis) in Mexico: implications for transfusion medicine in the United States
Authors: Kirchhoff Louis V. , Paredes Patricia , Lomelí‐Guerrero Abel , Paredes‐Espinoza Mario , Ron‐Guerrero Carlos S. , Delgado‐Mejía Manuel , Peña‐Muñoz José G.
Published: January 2006

BACKGROUND:  Trypanosoma cruzi, the protozoan cause of Chagas disease, causes life‐long infection and is easily transmitted by blood transfusion. Our goals were to determine the prevalence of Chagas disease among donors in five Mexican blood banks, to look for evidence of transmission of T. cruzi by transfusion, and to evaluate two serologic assays for Chagas disease.

STUDY DESIGN and METHODS:  Blood samples from donors were tested initially with the Abbott Chagas EIA or the Meridian Chagas’ IgG ELISA. Samples giving readings that were at least 50% of the cutoffs were run in a confirmatory radioimmune precipitation assay (RIPA), as were samples from recipients of blood products from RIPA‐positive donors.

RESULTS:  The overall prevalence of Chagas disease was 1/133 (55/7,296; 0.75%). In addition, 4 of 9 surviving recipients of blood products from T. cruzi‐infected donors were in turn infected. Using the manufacturers’ recommended cutoffs, the sensitivity and specificity of the Abbott test were 92.0% (23/25) and 99.8% (2,865/2,872) respectively, and the corresponding values for the Meridian assay were 70.0% (21/30) and 100.0% (4,369/4,369).

CONCLUSIONS:  These findings indicate clearly that transfusion‐associated transmission of T. cruzi is occurring in the study areas. Serologic testing of blood donors for Chagas disease should be performed there and in the rest of Mexico. The two screening assays evaluated may lack the accuracy necessary for blood donor testing when used as suggested by the manufacturers.
Comparison of two sterile connection devices and the effect of sterile connections on blood component quality
Authors: Van Der Meer Pieter F. , Eijzenga Marjan , Pietersz Ruby N.I.
Published: January 2006

BACKGROUND:  Sterile connection devices (SCDs) are used to connect pieces of polyvinylchloride tubing between blood bag systems. After observing a slight decrease in inner diameter of tubing welded with the CompoDock S2 SCD, the effect of welded tubing on storage characteristics of white blood cell (WBC)‐reduced red blood cells (RBCs) and platelet (PLT) concentrates was studied. Welds were made with Terumo SCD (T‐SCD) or CompoDock S2, and unwelded tubing served as reference.

STUDY DESIGN AND METHODS:  Three WBC‐reduced RBC units or 3 PLT concentrates were pooled and divided to prevent donor‐dependent differences. The units were transferred 10 times over (1) tubing with a T‐SCD weld, (2) a CompoDock S2 weld, or (3) unwelded tubing. RBCs were stored for 42 days and free hemoglobin (Hb) was measured; PLT concentrates were stored for 8 days and CD62P expression was measured, as markers for blood component quality (n = 10 paired experiments).

RESULTS:  WBC‐reduced RBC units had similar hemolysis at the end of storage: 0.47 ± 0.28, 0.47 ± 0.35, and 0.49 ± 0.38 percent of total Hb, for tubing with a T‐SCD weld, a CompoDock S2 weld, or no weld, respectively (not significant). CD62P expression of stored WBC‐reduced PLT concentrates was not significantly different between the groups: 20.3 ± 5.1, 19.8 ± 5.1, and 22.3 ± 9.8 percent for tubing with a T‐SCD weld, a CompoDock S2 weld, or no weld, respectively.

CONCLUSION:  The quality of blood components, measured as RBC hemolysis and platelet CD62P expression, is not adversely affected by the presence of a sterile connection in the tubing, made by either the CompoDock S2 or the T‐SCD.
A weak blood group A phenotype caused by a translation‐initiator mutation in the ABO gene
Authors: Seltsam Axel , Das Gupta Christa , Bade‐Doeding Christina , Blasczyk Rainer
Published: January 2006

BACKGROUND:  Weak blood group A and B phenotypes are correlated with ABO glycosyltransferases exhibiting single‐amino‐acid changes and/or C‐terminal modifications.

STUDY DESIGN AND METHODS:  A healthy donor diagnosed as having weak A antigen expression and his two children were subjected to extensive ABO typing. HeLa cells were used to transfect ABO expression plasmids.

RESULTS:  The donor's red blood cells were type AweakB and his serum sample contained weakly reactive anti‐A1 antibodies. A single T>C transition identified at the +2 position of the start codon of an ABO*A101 allele predicted the disruption of this methionine codon. In the transfection studies, a significant reduction of A activity was observed on HeLa cells transfected with a plasmid containing the variant ABO*A allele. Coexpression of the respective antithetical ABO*B101 wild‐type construct further reduced cell surface A antigen expression. Similar expression results were obtained with ABO constructs in which the Met1 start codon and five alternative start sites at codons 20, 26, 43, 53, and 69 had successively been interrupted.

CONCLUSION:  The donor's weak blood group A phenotype most likely resulted from expression of an N‐truncated A transferase triggered by alternative translation start sites in the transmembrane domain or stem region.
Mortality differences among organisms causing septicemia in hemodialysis patients
Authors: DANESE Mark D. , GRIFFITHS Robert I. , DYLAN Michelle , YU Hsing‐Ting , DUBOIS Robert , NISSENSON Allen R.
Published: January 2006

Septicemia is a serious problem in hemodialysis patients because it can lead to life‐threatening complications and a persistently elevated risk of death. Most analyses have not examined whether there are differences in mortality risk among the organisms that cause these episodes of septicemia. This study was a retrospective cohort analysis of first septicemia hospitalizations during the first year of hemodialysis. Time to death (both in‐hospital and within 12 weeks post‐discharge) was compared among the different septicemia‐causing organisms based on discharge diagnoses in Medicare billing data from 1996 to 2001. The effect of various complications on mortality risk was also evaluated. There were 22,130 septicemia hospitalizations identified. The most common organism identified was Staphylococcus aureus (27%), with no other organism having an incidence >10%. The overall unadjusted death rate from admission through 12 weeks of follow‐up was 34%. During the first hospitalization, the death rate was 14%, and during the 12‐week period after the hospitalization it was 20%. In adjusted analyses, S. aureus was associated with a 20% higher risk of death both during the in‐hospital period and the 12‐week post‐discharge period, when compared with all other specified organisms. Hospitalizations complicated by meningitis, stroke, or endocarditis were also associated with increased risk of mortality, independent of the organism causing septicemia. Septicemia hospitalizations are associated with a high mortality rate—both during the initial hospitalization and after discharge. Meningitis, stroke, and endocarditis represent particularly serious complications. Overall, septicemia hospitalizations (especially for S. aureus) are serious events, and patients would benefit from better treatment and prevention.
Extravascular lung water index: A new method to determine dry weight in chronic hemodialysis patients
Authors: KUHN Christian , KUHN Andrea , RYKOW Kai , OSTEN Bernd
Published: January 2006

To assess the dry weight of chronic hemodialysis (HD) patients, the extravascular lung water index (ELWI) as a volume parameter was investigated to identify fluid overload. Forty‐two patients (30 males, 12 females) with a mean age of 55.7±13.0 years who were clinically not overhydrated were connected to the PiCCO system before starting HD treatment. We determined ELWI (normal range 3–7 mL/kg) and the following parameters: global end‐diastolic volume index (GEDI, normal range 680–800 mL/m2) and intrathoracic blood volume index (ITBI, normal range 850–1000 mL/m2) before and after HD to assess the volume status. Brain natriuretic peptide (BNP), aldosterone, and renin as vasoactive hormones were measured at the beginning and at the end of HD treatment as well. In 28 of the 42 patients (67%), elevated values of ELWI were found, indicating interstitial volume overload. There were significant correlations between ELWI and cardiac function index (p=0.003; Pearson's coefficient−0.451), global ejection fraction (p=0.012; Pearson's coefficient −0.389), ITBI (p=0.004; Pearson's coefficient 0.437), and GEDI (p=0.004; Pearson's coefficient 0.437). No significant relations among ELWI and mean arterial pressure (MAP), BNP, aldosterone, and renin were found. In conclusion, the use of ELWI is safe in chronic HD patients and identifies fluid‐overloaded patients, who show no obvious signs of hypervolemia. The determination of ELWI is an excellent method to quantify the exact volume in chronic HD patients.
Radiotherapy of basal cell carcinoma of the face and head: Importance of low dose per fraction on long‐term outcome
Authors: Olschewski Thomas , Bajor Katharina , Lang Birgit , Lang Eugen , Seegenschmiedt Michael H.
Published: January 2006

Background: Radiotherapy plays an important role in the treatment of basal cell carcinoma of the face and head and achieves cure rates of 92‐96 %. Different fractionation concepts of radiotherapy have been described.This study investigates the efficacy, as well as acute and chronic toxicity, of a slightly hypofractionated radiotherapy schedule.

Patients and Methods: 85 patients with 104 tumors underwent radiotherapy for basal cell carcinoma of the face and head. The radiotherapy schedule was 5×3 Gray/week up to a total dose of 57 Gray in 95 % of patients. Acute and late radiotherapy toxicity and cosmetic outcome were evaluated in long‐term follow‐up.

Results: No recurrence was observed. In 87 % of tumors, only low acute toxicity occurred at the end of radiotherapy. Late toxicity, if observed, was low in most patients. “Excellent” or “good” cosmesis was achieved in 94 % of tumors at last follow‐up.

Conclusions: Our radiotherapy schedule achieves a very high local control rate and very good cosmetic and functional results.This fractionation can be recommended as a standardized radiotherapy treatment for basal cell carcinoma of the face and head.
A Readable Earlier Renaissance: Small Adjustments, Large Changes
Authors: Coldiron A. E. B.
Published: January 2006

“Readability” of several kinds is a problem in Renaissance or early modern literature. One way to enhance the readability of early modern texts is to adjust the (admittedly artificial) period boundary that begins the traditional “Renaissance.” Unlike the historically‐based boundaries of regnal years (1485, 1509), a slightly earlier date, 1476, implies a conception of “Renaissance” that takes as its pivotal starting point the establishment of the printing press in England. The earlier date thus refocuses our analysis of the period to systems of production, distribution, and reception that directly create and shape Renaissance literature. Such an adjustment also accomplishes a wider category challenge, since the literature of the early print period is qualitatively different from the more familiar literature of the Elizabethan and Jacobean periods. This essay explains that to shift the starting point of inquiry about “Renaissance” is to challenge the whole period concept and to invite new critical narratives about the period that will make it more “readable” in several senses (legible, historically comprehensible, lisible, literally accessible, etc.). The essay also reviews similar, recent, challenges to other critical concepts in early modern studies: to authorship, genre, theme, canon, method. Such challenges have been generative of greater “readability” and growth in the field.
Identification and characterization of 1‐Cys peroxiredoxin from Sulfolobus solfataricus and its involvement in the response to oxidative stress
Authors: Limauro Danila , Pedone Emilia , Pirone Luciano , Bartolucci Simonetta
Published: January 2006

Bcp2 was identified as a putative peroxiredoxin (Prx) in the genome database of the aerobic hyperthermophilic archaeon Sulfolobus solfataricus. Its role in oxidative stress was investigated by transcriptional analysis of RNA isolated from cultures that had been stressed with various oxidant agents. Its specific involvement was confirmed by a considerable increase in the bcp2 transcript following induction with H2O2. The 5′ end of the transcript was mapped by primer extension analysis and the promoter region was characterized. bcp2 was cloned and expressed in Escherichia coli, the recombinant enzyme was purified and the predicted molecular mass was confirmed. Using dithiothreitol as an electron donor, this enzyme acts as a catalyst in H2O2 reduction and protects plasmid DNA from nicking by the metal‐catalysed oxidation system. Western blot analysis revealed that the Bpc2 expression was induced as a cellular adaptation in response to the addition of exogenous stressors. The results obtained indicate that Bcp2 plays an important role in the peroxide‐scavaging system in S. solfataricus. Mutagenesis studies have shown that the only cysteine, Cys49, present in the Bcp2 sequence, is involved in the catalysis. Lastly, the presence of this Cys in the sequence confirms that Bcp2 is the first archaeal 1‐Cysteine peroxiredoxin (1‐Cys Prx) so far identified.
Proton‐decoupled 15N and 31P solid‐state NMR investigations of the Pf3 coat protein in oriented phospholipid bilayers
Authors: Aisenbrey Christopher , Harzer Ulrike , Bauer‐Manz Gabriele , Bär Gerda , Chotimah Irma N. Husnal , Bertani Philippe , Sizun Christina , Kuhn Andreas , Bechinger Burkhard
Published: January 2006

The coat proteins of filamentous phage are first synthesized as transmembrane proteins and then assembled onto the extruding viral particles. We investigated the transmembrane conformation of the Pseudomonas aeruginosa Pf3 phage coat protein using proton‐decoupled 15N and 31P solid‐state NMR spectroscopy. The protein was either biochemically purified and uniformly labelled with 15N or synthesized chemically and labelled at specific sites. The proteins were then reconstituted into oriented phospholipid bilayers and the resulting samples analysed. The data suggest a model in which the protein adopts a tilted helix with an angle of ≈ 30° and an N‐terminal ‘swinging arm’ at the membrane surface.
How we do it: The Owa (Addenbrookes) grommet
Authors: Bray D. , Owa A.
Published: January 2006

Keypoints

• Grommet insertion is one of the most common procedures performed under general anaesthesia.

• The authors describe an innovative grommet that, through an anatomically contoured and user friendly design has been designed to minimize anaesthetic time by facilitating and expediting its placement and/or removal.
A randomized trial of Rapid Rhino Riemann and Telfa nasal packs following endoscopic sinus surgery
Authors: Cruise A.S. , Amonoo‐Kuofi K. , Srouji I. , Kanagalingam J. , Georgalas C. , Patel N.N. , Badia L. , Lund V.J.
Published: January 2006

Objectives:  To compare Telfa with the Rapid Rhino Riemann nasal pack for use following endoscopic sinus surgery.

Design:  Prospective, randomized, double‐blind, paired trial.

Setting:  Tertiary otolaryngology hospital.

Participants:  Forty‐five adult patients undergoing bilateral endoscopic sinus surgery for either chronic rhinosinusitis or nasal polyps.

Main outcome measures:  A visual analogue scale was used to assess discomfort caused by the presence of the packs in the nose and by their removal. The amount of bleeding was noted with the packs in place and following their removal. Crusting and adhesions were assessed 2 and 6 weeks following surgery.

Results:  Both packs performed well giving good haemostasis and causing little bleeding on removal. Both packs caused only mild discomfort while in the nose. On the visual analogue scale of 0–10 cm the mean visual analogue score for Rapid Rhino Riemann pack was 1.7 and for Telfa 2.0 (P = 0.371). The Rapid Rhino Riemann pack caused significantly less pain on removal compared with the Telfa pack with a mean visual analogue score of 2.0 in comparison with 3.7 for Telfa (P = 0.001). There were less adhesions with the Rapid Rhino Riemann than Telfa pack but this was not statistically significant (P = 0.102).

Conclusions:  Both Telfa and Rapid Rhino Riemann packs can be recommended as packs that control postoperative haemorrhage, do not cause bleeding on removal and cause little discomfort while in the nose. The Rapid Rhino Riemann pack has the advantage of causing significantly less pain on removal.
Do chronic changes in nasal airflow have any physiological or pathological effect on the nose and paranasal sinuses? A systematic review
Authors: Boyce J. , Eccles R.
Published: January 2006

Keypoints

• A reduction in nasal airflow associated with anatomical defects of the nose such as nasal septal deviation has been proposed to cause nasal pathology.

• The majority of animal experiments where one nasal passage is surgically closed over several months report only minor changes in the histology of the nasal epithelium and no rhinitis or sinusitis.

• Complete abolition of nasal airflow associated with laryngectomy or the treatment of atrophic rhinitis is not associated with the development of rhinitis or sinusitis.

• Radiological studies have shown a lack of association between the degree of nasal septal deviation and evidence of rhinosinusitis.

• Such studies provide evidence that reduced nasal airflow causes no significant nasal disease.

• There is no convincing evidence that a reduction in nasal airflow is a causative factor for rhinitis or sinusitis.
Topical anaesthesia before nasendoscopy: a randomized controlled trial of co‐phenylcaine compared with lignocaine
Authors: Douglas R. , Hawke L. , Wormald P.‐J.
Published: January 2006

Objective:  To evaluate the relative effectiveness of co‐phenylcaine (lignocaine 5% with phenylephrine) and lignocaine 5% sprays when administered prior to rigid nasendoscopy.

Design:  Randomized, double blind controlled study.

Setting:  Teaching hospital otolaryngology unit.

Participants:  Thirty patients requiring routine outpatient rigid nasendoscopy were administered five puffs of either co‐phenylcaine or lignocaine 5% spray which had been randomly assigned to either the first or the second visit. Ten minutes later nasendoscopy was performed. Immediately after nasendoscopy the ease of performance of the procedure and the quality of the view achieved was rated on a visual analogue scale by the endoscopist and the patients recorded the level of pain experienced on a visual analogue scale. Two weeks later, the patients returned for a repeat nasendoscopy, receiving the alternate spray.

Main outcomes measures:  Ease of performance and quality of view of achieved by endoscopists and pain experienced by patients, both measured with visual analogue scales.

Results:  The ease of passage of the endoscope and quality of the view obtained was found to be greater after the administration of co‐phenylcaine [visual analogue scores 84 (95% CI: 80–89) than after lignocaine and 77 (95% CI: 73–81) (P < 0.01)]. The two sprays produced similar levels of topical anaesthesia.

Conclusions:  Nasendoscopy can be performed with minimal discomfort after the administration of either co‐phenylcaine or lignocaine 5% sprays. The vasoconstricting action of co‐phenylcaine increases the ease of passage of the endoscope and quality of the view obtained by the endoscopist.
Acoustic parameters of snoring sound to compare natural snores with snores during ‘steady‐state’ propofol sedation
Authors: Jones T.M. , Ho M.S. , Earis J.E. , Swift A.C. , Charters P.
Published: January 2006

Objectives:  To investigate the acoustic similarity between natural and sedation‐induced snores.

Design:  Prospective observational study.

Setting:  University Hospital Aintree, Liverpool, UK.

Participants:  Twenty‐one patients, who had already had overnight snore recordings, completed a pre‐operative sleep nasendoscopic examination. Endoscopic examination of the upper aero‐digestive tract was performed at sequentially increasing, steady‐state sedation levels, using intravenous propofol administered according to a weight/time‐based algorithm to predict blood and effect site (tissue) concentrations. At each sedation level at which snoring occurred, snoring sound was recorded. From these samples, snore files, comprising the inspiratory sound of each snore were created. Similarly, from natural snores recorded pre‐operatively, snore files, comprising the inspiratory sounds of the first 100 snores with the patient sleeping in a supine position, were also created.

Main outcome measures:  Snore duration (s), loudness (dBA), periodicity (%) and energy ratios for the frequency sub‐bands 0–200, 0–250 and 0–400 Hz.

Results:  Snore loudness increased significantly (P < 0.0001), whilst energy ratios for frequency bands 0–200, 0–250 and 0–400 Hz all decreased significantly as sedation level increased (P < 0.001). A significant difference between natural snoring and snoring induced at the lowest sedation level was shown (P < 0.0001). Endoscopic examination was not tolerated at this sedation level.

Conclusions:  The acoustic characteristics of sedation‐induced and natural snores are sufficiently different to recommend the need for further research to determine whether the technique of sleep nasendoscopy is, in fact, a valid predictor of outcome of snoring surgery.
How we do it: Septoplasties under local anaesthetic are suitable for short stay surgery; the clinical outcomes
Authors: Hytönen M. , Blomgren K. , Lilja M. , Mäkitie A.A.
Published: January 2006

Keypoints

• Septoplasties can be performed under local anaesthesia. However, careful prevention and treatment of pain during the operation is essential.

• Septoplasties are suitable operations for short‐stay surgery.

• Two‐years post‐operative, septoplasty relieved the symptoms well or excellently in 55% and moderately in 27% of the cases.

• Specialists prescribe more prophylactic antibiotics than residents (47%versus 29%P = 0.02).

• Post‐operative antibiotics do not appear to prevent the development of post‐operative infections.
Use of intravenous steroids at induction of anaesthesia for adult tonsillectomy to reduce post‐operative nausea and vomiting and pain: a double‐blind randomized controlled trial
Authors: McKean S. , Kochilas X. , Kelleher R. , Dockery M.
Published: January 2006

Objective:  To assess the effectiveness of intravenous steroids at induction of anaesthetic to reduce post‐operative nausea and vomiting and pain after adult tonsillectomy.

Design:  Prospective, double‐blind, randomized, placebo controlled trial, with ethical approval, following Consolidated Standards of Reporting Trials guidelines.

Setting:  District General Hospital in Scotland, UK.

Participants:  Seventy‐two adults between 16 and 70 years, American Association of Anaethetists (ASA) 1, listed for elective tonsillectomy.

Intervention:  Single dose of either 10 mg of dexamethasone or 2 mL of saline after induction with a consistent anaesthetic technique.

Main outcome measures:  Patients filled in a visual analogue scale relating to pain and post‐operative nausea and vomiting for the day of operation and 7 days after operation. The time to first ingestion of food and drink after operation was also noted.

Results:  Data completion rate of 64% (46 of 72 patients enrolled). Statistically significant relative decrease (62%P = 0.001) in the incidence of post‐operative nausea and vomiting was seen in those treated with dexamethasone. Statistically significant relative decrease (23%P = 0.016) in post‐operative pain scores for the day of operation was seen in those treated with dexamethasone. Significant decrease (17.5%, P < 0.001) in mean pain score for seven post‐operative days was seen in those treated with dexamethasone. No adverse effects were seen.

Conclusions:  Dexamethasone given as a single dose of 10 mg at induction of anaesthesia for adult tonsillectomy is an effective, safe and inexpensive method for reducing morbidity in adult tonsillectomy.
The Path Forward for Biofuels and Biomaterials
Authors: Ragauskas Arthur J. , Williams Charlotte K. , Davison Brian H. , Britovsek George , Cairney John , Eckert Charles A. , Frederick William J. , Hallett Jason P. , Leak David J. , Liotta Charles L. , Mielenz Jonathan R. , Murphy Richard , Templer Richard , Tschaplinski Timothy
Published: January 2006

Biomass represents an abundant carbon-neutral renewable resource for the production of bioenergy and biomaterials, and its enhanced use would address several societal needs. Advances in genetics, biotechnology, process chemistry, and engineering are leading to a new manufacturing concept for converting renewable biomass to valuable fuels and products, generally referred to as the biorefinery. The integration of agroenergy crops and biorefinery manufacturing technologies offers the potential for the development of sustainable biopower and biomaterials that will lead to a new manufacturing paradigm.
An Architectural Framework That May Lie at the Core of the Postsynaptic Density
Authors: Baron Marisa K. , Boeckers Tobias M. , Vaida Bianca , Faham Salem , Gingery Mari , Sawaya Michael R. , Salyer Danielle , Gundelfinger Eckart D. , Bowie James U.
Published: January 2006

The postsynaptic density (PSD) is a complex assembly of proteins associated with the postsynaptic membrane that organizes neurotransmitter receptors, signaling pathways, and regulatory elements within a cytoskeletal matrix. Here we show that the sterile alpha motif domain of rat Shank3/ProSAP2, a master scaffolding protein located deep within the PSD, can form large sheets composed of helical fibers stacked side by side. Zn
2+
, which is found in high concentrations in the PSD, binds tightly to Shank3 and may regulate assembly. Sheets of the Shank protein could form a platform for the construction of the PSD complex.
Rapid Uplift of the Altiplano Revealed Through
13
C-
18
O Bonds in Paleosol Carbonates
Authors: Ghosh Prosenjit , Garzione Carmala N. , Eiler John M.
Published: January 2006

The elevation of Earth's surface is among the most difficult environmental variables to reconstruct from the geological record. Here we describe an approach to paleoaltimetry based on independent and simultaneous determinations of soil temperatures and the oxygen isotope compositions of soil waters, constrained by measurements of abundances of
13
C-
18
O bonds in soil carbonates. We use this approach to show that the Altiplano plateau in the Bolivian Andes rose at an average rate of 1.03 ± 0.12 millimeters per year between ∼10.3 and ∼6.7 million years ago. This rate is consistent with the removal of dense lower crust and/or lithospheric mantle as the cause of elevation gain.
Comment on "Reconstructing the Origin of Andaman Islanders"
Authors: Palanichamy Malliya gounder , Agrawal Suraksha , Yao Yong-Gang , Kong Qing-Peng , Sun Chang , Khan Faisal , Chaudhuri Tapas Kumar , Zhang Ya-Ping
Published: January 2006

On the basis of mitochondrial DNA sequence analyses, Thangaraj
et al
. (Brevia, 13 May 2005, p. 996) proposed that Andaman islanders descended from the first humans to migrate out of Africa. We identified mitochondrial DNA from two northeast Indian Rajbanshi individuals that shares three specific mutations with the M31a lineage observed in the Great Andamanese, which suggests that the predecessor of haplogroup M31 originated on the Indian subcontinent.
Community Genomics Among Stratified Microbial Assemblages in the Ocean's Interior
Authors: DeLong Edward F. , Preston Christina M. , Mincer Tracy , Rich Virginia , Hallam Steven J. , Frigaard Niels-Ulrik , Martinez Asuncion , Sullivan Matthew B. , Edwards Robert , Brito Beltran Rodriguez , Chisholm Sallie W. , Karl David M.
Published: January 2006

Microbial life predominates in the ocean, yet little is known about its genomic variability, especially along the depth continuum. We report here genomic analyses of planktonic microbial communities in the North Pacific Subtropical Gyre, from the ocean's surface to near–sea floor depths. Sequence variation in microbial community genes reflected vertical zonation of taxonomic groups, functional gene repertoires, and metabolic potential. The distributional patterns of microbial genes suggested depth-variable community trends in carbon and energy metabolism, attachment and motility, gene mobility, and host-viral interactions. Comparative genomic analyses of stratified microbial communities have the potential to provide significant insight into higher-order community organization and dynamics.
Believing in Belief
Authors: Shermer Michael
Published: January 2006

Breaking the Spell
. Religion as a Natural Phenomenon.
By Daniel C. Dennett
. Viking, New York, 2006. 464 pp. $25.95, C$36. ISBN 0-670-03472-X.

The author draws on philosophy, psychology, and history to consider the source and purpose of human devotion to gods.
Envelope Determinants for Dual-Receptor Specificity in Feline Leukemia Virus Subgroup A and T Variants
Authors: Cheng Heather H. , Anderson Maria M. , Hankenson F. Claire , Johnston Lily , Kotwaliwale Chitra V. , Overbaugh Julie
Published: January 2006

Gammaretroviruses, including the subgroups A, B, and C of feline leukemia virus (FeLV), use a multiple-membrane-spanning transport protein as a receptor. In some cases, such as FeLV-T, a nonclassical receptor that includes both a transport protein (Pit1) and a soluble cofactor (FeLIX) is required for entry. To define which regions confer specificity to classical versus nonclassical receptor pathways, we engineered mutations found in either FeLV-A/T or FeLV-T, individually and in combination, into the backbone of the transmissible form of the virus, FeLV-A. The receptor specificities of these viruses were tested by measuring infection and binding to cells expressing the FeLV-A receptor or the FeLV-T receptors. FeLV-A receptor specificity was maintained when changes at amino acid position 6, 7, or 8 of the mature envelope glycoprotein were introduced, although differences in infection efficiency were observed. When these N-terminal mutations were introduced together with a C-terminal 4-amino-acid insertion and an adjacent amino acid change, the resulting viruses acquired FeLV-T receptor specificity. Additionally, a W→L change at amino acid position 378, although not required, enhanced infectivity for some viruses. Thus, we have found that determinants in the N and C termini of the envelope surface unit can direct entry via the nonclassical FeLV-T receptor pathway. The region that has been defined as the receptor binding domain of gammaretroviral envelope proteins determined entry via the FeLV-A receptor independently of the presence of the N- and C-terminal FeLV-T receptor determinants.
Structure-Function Analysis of the Epitope for 4E10, a Broadly Neutralizing Human Immunodeficiency Virus Type 1 Antibody
Authors: Brunel Florence M. , Zwick Michael B. , Cardoso Rosa M. F. , Nelson Josh D. , Wilson Ian A. , Burton Dennis R. , Dawson Philip E.
Published: January 2006

The human immunodeficiency virus type 1 (HIV-1) neutralizing antibody 4E10 binds to a linear, highly conserved epitope within the membrane-proximal external region of the HIV-1 envelope glycoprotein gp41. We have delineated the peptide epitope of the broadly neutralizing 4E10 antibody to gp41 residues 671 to 683, using peptides with different lengths encompassing the previously suggested core epitope (NWFDIT). Peptide binding to the 4E10 antibody was assessed by competition enzyme-linked immunosorbent assay, and theKdvalues of selected peptides were determined using surface plasmon resonance. An Ala scan of the epitope indicated that several residues, W672, F673, and T676, are essential (>1,000-fold decrease in binding upon replacement with alanine) for 4E10 recognition. In addition, five other residues, N671, D674, I675, W680, and L679, make significant contributions to 4E10 binding. In general, the Ala scan results agree well with the recently reported crystal structure of 4E10 in complex with a 13-mer peptide and with our circular dichroism analyses. Neutralization competition assays confirmed that the peptide NWFDITNWLWYIKKKK-NH2could effectively inhibit 4E10 neutralization. Finally, to limit the conformational flexibility of the peptides, helix-promoting 2-aminoisobutyric acid residues and helix-inducing tethers were incorporated. Several peptides have significantly improved affinity (>1,000-fold) over the starting peptide and, when used as immunogens, may be more likely to elicit 4E10-like neutralizing antibodies. Hence, this study represents the first stage toward iterative development of a vaccine based on the 4E10 epitope.
Comprehensive Mapping of Receptor-Functioning Domains in Feline Leukemia Virus Subgroup C Receptor FLVCR1
Authors: Brown Jennifer K. , Fung Claire , Tailor Chetankumar S.
Published: January 2006

Infection of cells by the highly anemogenic feline leukemia virus subgroup C (FeLV-C) is mediated by the heme exporter FLVCR1, a cell surface protein containing 12 potential transmembrane segments with six presumptive extracellular loops (ECLs). To identify FLVCR1 residues critical for mediating FeLV-C infection, we first independently isolated a human cDNA encoding the FLVCR2 protein that shares 52% identity to human FLVCR1, and we show that FLVCR2 does not function as a receptor for FeLV-C. Then, by generating specific hybrids between FLVCR1 and FLVCR2 and testing susceptibility of mouse cells expressing these hybrids to β-galactosidase encoding FeLV-C, we identify FLVCR1 ECLs 1 and 6 as critical for mediating FeLV-C infection. Mouse cells expressing a hybrid protein containing FLVCR2 backbone with the ECL6 sequence from FLVCR1 were highly susceptible to FeLV-C infection. Using site-directed mutagenesis, we show that a single mutation of Asn463 in FLVCR2 ECL6 to an acidic Asp residue (a residue present in the corresponding position 487 in FLVCR1 ECL6) is sufficient to render FLVCR2 functional as an FeLV-C receptor. However, an Asp487Asn mutation in FLVCR1 ECL6 or substitution of the entire FLVCR1 ECL6 sequence for FLVCR2 ECL6 sequence does not disrupt receptor function. Subsequent substitutions show that residues within FLVCR1 ECL1 also contribute to mediating FeLV-C infection. Furthermore, our results suggest that FLVCR1 regions that mediate FeLV-C surface unit binding are distinct from ECL1 and ECL6. Our results are consistent with previous conclusions that infection of cells by gammaretroviruses involves interaction of virus with multiple receptor regions.
Rectal Immunization with Rotavirus Virus-Like Particles Induces Systemic and Mucosal Humoral Immune Responses and Protects Mice against Rotavirus Infection
Authors: Parez Nathalie , Fourgeux Cynthia , Mohamed Ali , Dubuquoy Catherine , Pillot Mathieu , Dehee Axelle , Charpilienne Annie , Poncet Didier , Schwartz-Cornil Isabelle , Garbarg-Chenon Antoine
Published: January 2006

To evaluate whether the rectal route of immunization may be used to provide appropriate protection against enteric pathogens such as rotaviruses (RV), we studied the antibody response and the protection induced by rectal immunization of mice with RV virus-like particles (VLP). For this purpose, 6-week-old BALBc mice were rectally immunized twice with RV 8-2/6/7-VLP derived from the bovine RV RF81 strain either alone or combined with various adjuvants including four toxins [cholera toxin (CT) and three attenuated
Escherichia coli
-derived heat-labile toxins (LTs), LT(R192G), LT(R72), and LT(K63)] and two Toll-like receptor-targeting adjuvants (CpG and resiquimod). Six weeks after the second immunization, mice were challenged with murine RV strain ECw. RV VLP administered alone were not immunogenic and did not protect mice against RV challenge. By contrast, RV VLP combined with any of the toxin adjuvants were immunogenic (mice developed significant titers of anti-RV immunoglobulin A [IgA] in both serum and feces and of anti-RV IgG in serum) and either efficiently induced complete protection of the mice (no detectable fecal virus shedding) or, for LT(K63), reduced the amount of fecal virus shedding after RV challenge. When combined with RV VLP, CpG and resiquimod failed to achieve protection, although CpG efficiently induced an antibody response to RV. These results support the consideration of the rectal route for the development of new immunization strategies against RV infection. Rectal delivery of a VLP-based vaccine might allow the use of adjuvants less toxic than, but as efficient as, CT.
Transcriptome Signature of Virulent and Attenuated Pseudorabies Virus-Infected Rodent Brain
Authors: Paulus Christina , Sollars Patricia J. , Pickard Gary E. , Enquist Lynn W.
Published: January 2006

Mammalian alphaherpesviruses normally establish latent infections in ganglia of the peripheral nervous system in their natural hosts. Occasionally, however, these viruses spread to the central nervous system (CNS), where they cause damaging, often fatal, infections. Attenuated alphaherpesvirus derivatives have been used extensively as neuronal circuit tracers in a variety of animal models. Their circuit-specific spread provides a unique paradigm to study the local and global CNS response to infection. Thus, we systematically analyzed the host gene expression profile after acute pseudorabies virus (PRV) infection of the CNS using Affymetrix GeneChip technology. Rats were injected intraocularly with one of three selected virulent and attenuated PRV strains. Relative levels of cellular transcripts were quantified from hypothalamic and cerebellar tissues at various times postinfection. The number of cellular genes responding to infection correlated with the extent of virus dissemination and relative virulence of the PRV strains. A total of 245 out of 8,799 probe sets, corresponding to 182 unique cellular genes, displayed increased expression ranging from 2- to more than 100-fold higher than in uninfected tissue. Over 60% thereof were categorized as immune, proinflammatory, and other cellular defense genes. Additionally, a large fraction of infection-induced transcripts represented cellular stress responses, including glucocorticoid- and redox-related pathways. This is the first comprehensive in vivo analysis of the global transcriptional response of the mammalian CNS to acute alphaherpesvirus infection. The differentially regulated genes reported here are likely to include potential diagnostic and therapeutic targets for viral encephalitides and other neurodegenerative or neuroinflammatory diseases.
Hyperphosphorylation of the Rotavirus NSP5 Protein Is Independent of Serine 67 or NSP2, and the Intrinsic Insolubility of NSP5 Is Regulated by Cellular Phosphatases
Authors: Sen Adrish , Agresti Darin , Mackow Erich R.
Published: January 2006

The NSP5 protein is required for viroplasm formation during rotavirus infection and is hyperphosphorylated into 32- to 35-kDa isoforms. Earlier studies reported that NSP5 is not hyperphosphorylated without NSP2 coexpression or deleting the NSP5 N terminus and that serine 67 is essential for NSP5 hyperphosphorylation. In this report, we show that full-length NSP5 is hyperphosphorylated in the absence of NSP2 or serine 67 and demonstrate that hyperphosphorylated NSP5 is predominantly present in previously unrecognized cellular fractions that are insoluble in 0.2% sodium dodecyl sulfate. The last 68 residues of NSP5 are sufficient to direct green fluorescent protein into insoluble fractions and cause green fluorescent protein localization into viroplasm-like structures; however, NSP5 insolubility was intrinsic and did not require NSP5 hyperphosphorylation. When we mutated serine 67 to alanine we found that the NSP5 mutant was both hyperphosphorylated and insoluble, identical to unmodified NSP5, and as a result serine 67 is not required for NSP5 phosphorylation. Interestingly, treating cells with the phosphatase inhibitor calyculin A permitted the accumulation of soluble hyperphosphorylated NSP5 isoforms. This suggests that soluble NSP5 is constitutively dephosphorylated by cellular phosphatases and demonstrates that hyperphosphorylation does not direct NSP5 insolubility. Collectively these findings indicate that NSP5 hyperphosphorylation and insolubility are completely independent parameters and that analyzing insoluble NSP5 is essential for studies assessing NSP5 phosphorylation. Our results also demonstrate the involvement of cellular phosphatases in regulating NSP5 phosphorylation and indicate that in the absence of other rotavirus proteins, domains on soluble and insoluble NSP5 recruit cellular kinases and phosphatases that coordinate NSP5 hyperphosphorylation.
Core Protein of Pestiviruses Is Processed at the C Terminus by Signal Peptide Peptidase
Authors: Heimann Manuela , Sosa Gleyder Roman , Martoglio Bruno , Thiel Heinz-Jürgen , Rümenapf Till
Published: January 2006

The core protein of pestiviruses is released from the polyprotein by viral and cellular proteinases. Here we report on an additional intramembrane proteolytic step that generates the C terminus of the core protein. C-terminal processing of the core protein of classical swine fever virus (CSFV) was blocked by the inhibitor (Z-LL)
2
-ketone, which is specific for signal peptide peptidase (SPP). The same effect was obtained by overexpression of the dominant-negative SPP D
265
A mutant. The presence of (Z-LL)
2
-ketone reduced the viability of CSFV almost 100-fold in a concentration-dependent manner. Reduction of virus viability was also observed in infection experiments using a cell line that inducibly expressed SPP D
265
A. The position of SPP cleavage was determined by C-terminal sequencing of core protein purified from virions. The C terminus of CSFV core protein is alanine
255
and is located in the hydrophobic center of the signal peptide. The intramembrane generation of the C terminus of the CSFV core protein is almost identical to the processing scheme of the core protein of hepatitis C viruses.
Human Immunodeficiency Virus Type 1 Incorporated with Fusion Proteins Consisting of Integrase and the Designed Polydactyl Zinc Finger Protein E2C Can Bias Integration of Viral DNA into a Predetermined Chromosomal Region in Human Cells
Authors: Tan Wenjie , Dong Zheng , Wilkinson Thomas A. , Barbas Carlos F. , Chow Samson A.
Published: January 2006

In vitro studies using fusion proteins consisting of human immunodeficiency virus type 1 integrase (IN) and a synthetic polydactyl zinc finger protein E2C, a sequence-specific DNA-binding protein, showed that integration of retroviral DNA can be biased towards a contiguous 18-bp E2C-recognition site. To determine whether the fusion protein strategy can achieve site-specific integration in vivo, viruses were prepared by cotransfection and various IN-E2C fusion proteins were packaged in
trans
into virions. The resulting viruses incorporated with the IN-E2C fusion proteins were functional and capable of performing integration at a level ranging from 1 to 24% of that of viruses containing wild-type (WT) IN. Two of the more infectious viruses, which contained E2C fused to either the N (E2C/IN) or to the C (IN/E2C) terminus of IN, were tested for their ability to direct integration into a unique E2C-binding site present within the 5′ untranslated region of
erbB-2
gene on human chromosome 17. The copy number of proviral DNA was measured using a quantitative real-time nested-PCR assay, and the specificity of directed integration was determined by comparing the number of proviruses within the vicinity of the E2C-binding site to that in the whole genome. Viruses containing IN/E2C fusion proteins had sevenfold higher preference for integrating near the E2C-binding site than those viruses containing WT IN, whereas viruses containing E2C/IN had 10-fold higher preference. The results indicated that the IN-E2C fusion protein strategy is capable of directing integration of retroviral DNA into a predetermined chromosomal region in the human genome.
Protection of Mice and Poultry from Lethal H5N1 Avian Influenza Virus through Adenovirus-Based Immunization
Authors: Gao Wentao , Soloff Adam C. , Lu Xiuhua , Montecalvo Angela , Nguyen Doan C. , Matsuoka Yumi , Robbins Paul D. , Swayne David E. , Donis Ruben O. , Katz Jacqueline M. , Barratt-Boyes Simon M. , Gambotto Andrea
Published: January 2006

The recent emergence of highly pathogenic avian influenza virus (HPAI) strains in poultry and their subsequent transmission to humans in Southeast Asia have raised concerns about the potential pandemic spread of lethal disease. In this paper we describe the development and testing of an adenovirus-based influenza A virus vaccine directed against the hemagglutinin (HA) protein of the A/Vietnam/1203/2004 (H5N1) (VN/1203/04) strain isolated during the lethal human outbreak in Vietnam from 2003 to 2005. We expressed different portions of HA from a recombinant replication-incompetent adenoviral vector, achieving vaccine production within 36 days of acquiring the virus sequence. BALB/c mice were immunized with a prime-boost vaccine and exposed to a lethal intranasal dose of VN/1203/04 H5N1 virus 70 days later. Vaccination induced both HA-specific antibodies and cellular immunity likely to provide heterotypic immunity. Mice vaccinated with full-length HA were fully protected from challenge with VN/1203/04. We next evaluated the efficacy of adenovirus-based vaccination in domestic chickens, given the critical role of fowl species in the spread of HPAI worldwide. A single subcutaneous immunization completely protected chickens from an intranasal challenge 21 days later with VN/1203/04, which proved lethal to all control-vaccinated chickens within 2 days. These data indicate that the rapid production and subsequent administration of recombinant adenovirus-based vaccines to both birds and high-risk individuals in the face of an outbreak may serve to control the pandemic spread of lethal avian influenza.