bluuebunny/crossref_metadata_2025 huggingface dataset:
Using the Proteomics Identifications Database (PRIDE)
Authors: Martens Lennart , Jones Phil , Côté Richard
Published: March 2008

The Proteomics Identifications Database (PRIDE) is a public data repository designed to store, disseminate, and analyze mass spectrometry based proteomics datasets. The PRIDE database can accommodate any level of detailed metadata about the submitted results, which can be queried, explored, viewed, or downloaded via the PRIDE Web interface. The PRIDE database also provides a simple, yet powerful, access control mechanism that fully supports confidential peer‐reviewing of data related to a manuscript, ensuring that these results remain invisible to the general public while allowing referees and journal editors anonymized access to the data. This unit describes in detail the functionality that PRIDE provides with regards to searching, viewing, and comparing the available data, as well as different options for submitting data to PRIDE. Curr. Protoc. Bioinform. 21:13.8.1‐13.8.19. © 2008 by John Wiley & Sons, Inc.
Using GFS to Identify Encoding Genomic Loci from Protein Mass Spectral Data
Authors: Holmes Mark R. , Giddings Morgan C.
Published: March 2008

Genome‐based peptide fingerprint scanning (GFS) directly maps several types of protein mass spectral (MS) data to the loci in the genome that may have encoded for the protein. This process can be used either for protein identification or for proteogenomic mapping, which is gene‐finding and annotation based on proteomic data. Inputs to the program are one or more mass spectrometry files from peptide mass fingerprinting and/or tandem MS (MS/MS) along with one or more sequences to search them against, and the output is the coordinates of any matches found. This unit describes the use of GFS and subsequent results analysis. Curr. Protoc. Bioinform. 21:13.9.1‐13.9.20. © 2008 by John Wiley & Sons, Inc.
Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells
Authors: Gonzalez Rodolfo , Loring Jeanne F. , Snyder Evan Y.
Published: March 2008

Human embryonic stem cells (hESCs) that are currently distributed under NIH guidelines, as well as many of those that are not on the NIH registry, have been derived and maintained in coculture with growth‐arrested mouse embryonic fibroblasts (MEFs). Using this mouse support system may compromise the therapeutic potential of these hESCs because of the risk of transmitting xenopathogens. Alternatively, to reduce this risk, methods to culture undifferentiated hESCs on autologous hESC‐derived human feeder layers have now been developed. This feeder cell system derived from hESCs successfully prolongs growth of undifferentiated hESCs and eliminates risk factors and concerns about using xenogeneic or unknown allogeneic feeders. In this unit, we provide the necessary protocols for an autogeneic human feeder system that efficiently supports hESC growth and maintenance of pluripotency. Curr. Protoc. Stem Cell Biol. 4:1C.5.1‐1C.5.15. © 2008 by John Wiley & Sons, Inc.
Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
Authors: Crisan Mihaela , Huard Johnny , Zheng Bo , Sun Bin , Yap Solomon , Logar Alison , Giacobino Jean‐Paul , Casteilla Louis , Péault Bruno
Published: March 2008

Multilineage progenitor cells, diversely designated as MSC, MAPC, or MDSC, have been previously extracted from long‐term cultures of fetal and adult organs (e.g., bone marrow, brain, lung, pancreas, muscle, adipose tissue, and several others). The identity and location, within native tissues, of these elusive stem cells are described here. Subsets of endothelial cells and pericytes, which participate in the architecture of human blood vessels, exhibit, following purification to homogeneity, developmental multipotency. The selection from human tissues, by flow cytometry using combinations of positive and negative cell surface markers, of endothelial and perivascular cells is described here. In addition, a rare subset of myoendothelial cells that express markers of both endothelial and myogenic cell lineages and exhibit dramatic myogenic and cardiomyogenic potential has been identified and purified from skeletal muscle. The culture conditions amenable to the long‐term proliferation of these blood vessel–associated stem cells in vitro are also described. Curr. Protoc. Stem Cell Biol. 4:2B.2.1‐2B.2.13. © 2008 by John Wiley & Sons, Inc.
Rural adult education: Current status
Authors: Ritchey Jeffrey A.
Published: March 2008

This chapter provides a brief overview of the term rural and a summary of critical issues important to adult and continuing educators working with rural populations.
Rural adult literacy in a community context: From the margin to the mainstream
Authors: Ziegler Mary F. , Davis Dent C.
Published: March 2008

Legislation over the past two decades has shaped the dominant view of adult literacy as addressing gaps in well‐defined, measurable skills. As rural communities face unprecedented challenges, this one‐size‐fits‐all view of adult literacy does not consider the unique local context of rural communities or involve those communities in defining their own needs and goals.
University and community collaborations in migrant ESL
Authors: McLaughlin John , Rodriguez Maria , Madden Carolyn
Published: March 2008

This chapter describes a university‐based migrant farmworker outreach and education program in Michigan and the difference it has made in the rural community it serves, as well as in the lives of the university students who have participated in it.
Defining rural community(ies): Future considerations for informal and nonformal adult education in rural communities
Authors: Bracken Susan J.
Published: March 2008

This chapter examines ways that rural character or community can be defined and suggests that future rural adult education research should be framed to include more specific considerations of rural context.
Rural adult education: Future directions
Authors: Ritchey Jeffrey A.
Published: March 2008

This chapter looks broadly at the preceding chapters, offering some thematic analysis as well as proposing possible areas for additional research.
A nasal proteosome adjuvant activates microglia and prevents amyloid deposition
Authors: Frenkel Dan , Puckett Lindsay , Petrovic Sanja , Xia Weiming , Chen Guiquan , Vega Jose , Dembinsky‐Vaknin Adi , Shen Jie , Plante Martin , Burt David S. , Weiner Howard L.
Published: March 2008

Objective

We assessed whether peripheral activation of microglia by a nasal proteosome‐based adjuvant (Protollin) that has been given safely to humans can prevent amyloid deposition in young mice and affect amyloid deposition and memory function in old mice with a large amyloid load.

Methods

Amyloid precursor protein (APP) transgenic (Tg) J20 mice received nasal treatment with Protollin weekly for 8 months beginning at age 5 months. Twenty‐four‐month‐old J20 mice were treated weekly for 6 weeks.

Results

We found reduction in the level of fibrillar amyloid (93%), insoluble β‐amyloid (Aβ; 68%), and soluble Aβ (45%) fragments in 14‐month‐old mice treated with Protollin beginning at age 5 months. Twenty‐four‐month‐old mice treated with nasal Protollin for 6 weeks had decreased soluble and insoluble Aβ (1‐40) and (1‐42) and improved memory function. Activated microglia (CD11b+ cells) colocalized with Aβ fibrils in the 24‐month‐old animals, and microglial activation correlated with the decrease in Aβ. No microglial activation was observed in 14‐month‐old mice, suggesting that once Aβ is cleared, there is downregulation of microglial activation. Both groups had reduction in astrocytosis. Protollin was observed in the nasal cavity and cervical lymph node but not in the brain. Activated CD11b+SRA+ (scavenger receptor A) cells were found in blood and cervical lymph node and increased interleukin‐10 in cervical lymph node. No toxicity was associated with treatment.

Interpretation

Our results demonstrate a novel antibody‐independent immunotherapy for both prevention and treatment of Alzheimer's disease that is mediated by peripheral activation of microglia with no apparent toxicity. Ann Neurol 2008;63:591–601
Fatal familial insomnia: Clinical features and early identification
Authors: Krasnianski Anna , Bartl Mario , Sanchez Juan Pascual J. , Heinemann Uta , Meissner Bettina , Varges Daniela , Schulze‐Sturm Ulf , Kretzschmar Haus A. , Schulz‐Schaeffer Walter J. , Zerr Inga
Published: March 2008

Our aim was to develop a detailed clinical description of fatal familial insomnia in a large patient group with respect to the M129V genotype. Data on 41 German fatal familial insomnia patients were analyzed. Clinical features, 14‐3‐3 proteins in the cerebrospinal fluid, magnetic resonance imaging, positron emission tomography, single‐photon emission computed tomography, polysomnography, and electroencephalography were studied. Age at disease onset, disease duration, and clinical syndrome varied depending on the codon 129 genotype. Because the sensitivity of the most diagnostic tests is low in fatal familial insomnia, detailed clinical investigation is extremely important. Polysomnography may help to support the diagnosis. Ann Neurol 2008
Effects of depth and eutrophication on the zooxanthella clades of Stylophora pistillata from the Gulf of Eilat (Red Sea)
Authors: Lampert‐Karako Sarit , Stambler Noga , Katcoff Don J. , Achituv Yair , Dubinsky Zvy , Simon‐Blecher Noa
Published: March 2008

The symbiotic association between corals and zooxanthellae has been a major contributing factor in the success of reef‐building corals.

The diversity of zooxanthellae harboured in the hermatypic coral Stylophora pistillata in the Gulf of Eilat (Aqaba), northern Red Sea, was investigated on the basis of the length and sequence variation of the 18S small subunit ribosomal DNA. Of several clades found worldwide, only clades A and C were found to be associated with this coral species in the Gulf.

No correlation between the pink and brown colour morphs of the coral S. pistillata and the zooxanthella clade was found. Shallow‐water colonies (5 m) harbour clade A while deeper‐water colonies (17 m) harbour either clade A or C. This coral is a brooding species that associates with different zooxanthellae at different depths.

Eutrophication from fish farms did not affect clade identity associated with Stylophora.

Copyright © 2008 John Wiley & Sons, Ltd.
Spatio‐temporal modeling of nanoparticle delivery to multicellular tumor spheroids
Authors: Goodman Thomas T. , Chen Jingyang , Matveev Konstantin , Pun Suzie H.
Published: March 2008

The inefficiency of nanoparticle penetration in tissues limits the therapeutic efficacy of such formulations for cancer applications. Recent work has indicated that modulation of tissue architecture with enzymes such as collagenase significantly increases macromolecule delivery. In this study we developed a mathematical model of nanoparticle penetration into multicellular spheroids that accounts for radially dependent changes in tumor architecture, as represented by the volume fraction of tissue accessible to nanoparticle diffusion. Parameters such as nanoparticle binding, internalization rate constants, and accessible volume fraction were determined experimentally. Unknown parameters of nanoparticle binding sites per cell in the spheroid and pore shape factor were determined by fitting to experimental data. The model was correlated with experimental studies of the penetration of 40 nm nanoparticles in SiHa multicellular spheroids with and without collagenase treatment and was able to accurately predict concentration profiles of nanoparticles within spheroids. The model was also used to investigate the effects of nanoparticle size. This model contributes toward the understanding of the role of tumor architecture on nanoparticle delivery efficiency. Biotechnol. Bioeng. 2008;101: 388–399. © 2008 Wiley Periodicals, Inc.
Stent fracture, an incidental finding or a significant marker of clinical in‐stent restenosis?
Authors: Shaikh Fareed , Maddikunta Rajesh , Djelmami‐Hani Mohamed , Solis Joaquin , Allaqaband Suhail , Bajwa Tanvir
Published: March 2008

Background:

The predictors and clinical significance for stent fracture (SF) in drug‐eluting stents (DES) remain unknown. We identified procedural factors leading to SF and its clinical consequences in DES.

Methods:

Percutaneous coronary interventions were performed on 3,920 patients with DES over 12 months. In‐stent restenosis (ISR) of DES was observed in 188 cases with 121 cases (64.4%) receiving a sirolimus‐eluting stent (SES) and 67 (35.6%) a paclitaxol‐eluting stent (PES).

Results:

SF was identified in 35 (18.6%) of the 188 cases. The 35 cases were then compared with 153 cases of ISR without angiographic evidence of SF. SF was identified in 29 (23.9%) SES compared with 6 (9.0%) in PES (P < 0.05). With univariate analysis, additional factors associated with SF included longer mean stented segment length, male gender, overlapping stents, vessel segment angulation >75°, and more stents (all P < 0.05). With multivariate adjustment, three factors, i.e., stenting on a bend >75° (OR = 13.8, 95%CI 3.7 to 51; P < 0.001), SES (OR = 4.1, 95%CI 1.3 to 13.4; P < 0.018) and overlapping stented segments (OR = 3.9, 95%CI 1.1 to 14.1; P < 0.041) were statistically significant independent predictors of SF while larger stent diameter was protective (OR = 0.14, 95%CI 0.04 to 0.70; P < 0.017).

Conclusion:

SF proved to be associated with angiographically‐documented clinical ISR. Although the exact mechanism is unknown, factors that appear to play a negative role in SF include vessel tortuosity, use of SES and overlapping stents. Larger stent diameter was protective. Further studies are needed to better define the factors important in the mechanism of SF. © 2008 Wiley‐Liss, Inc.
Survival advantage in Medicare patients receiving drug‐eluting stents compared with bare metal stents: Real or artefactual?
Authors: Wang Fen Wei , Uretsky Barry F. , Freeman Jean L. , Zhang Dong , Giordano Sharon H. , Goodwin James S.
Published: March 2008

Background:

Concerns have been raised regarding late mortality, particularly from late stent thrombosis, from drug‐eluting stents (DES). Randomized clinical trials have shown that DES decrease restenosis but do not decrease mortality compared with bare metal stents (BMS). These studies utilized well‐defined clinical and angiographic subsets. In the “real world” drug‐eluting stents are used in a much broader crosssection of patients. We evaluated mortality in the first year after implantation of DES, specifically the sirolimus‐eluting stent (SES), Cypher vs. BMS in “real world” older patients using the Medicare claims database.

Methods and Results:

Data for the years 2002 (n = 6,890; pre‐DES) and 2003 (n = 7,566; first year of DES use) (May through December of each year) were analyzed. BMS and DES groups had similar baseline characteristics except for small but significant differences with BMS patients being somewhat older, having more males and African Americans, and a higher percentage of peripheral artery disease and heart failure while DES patients had a higher percentage of diabetics and patients with prior revascularization procedures. A significant improvement in mortality using both unadjusted and adjusted analyses was observed for DES (6.0% vs. 11.4%, P < 0.0001; hazard ratio 1.98, 95% CI 1.68–2.34). Controlling for comorbidity, extent of disease, and other chararacteristics by multivariable analysis or by propensity analysis had little impact on these results. On the other hand, there was no change in overall mortality in all stented patients in 2003 compared with all stented patients in 2002.

Conclusion:

An observed mortality benefit for DES compared with BMS in 2003 was observed, demonstrating the safety of DES, and suggesting the possibility of superiority in outcome in older patients with DES vs. BMS. However, the lack of improved survival from 2002 to 2003 in all stented patients suggests that the mortality advantage with DES finding may be due to unidentified selection biases. Our data suggest that DES in the Medicare population is as safe as, and possibly superior, to BMS for survival over the first year after implantation. © 2008 Wiley‐Liss, Inc.
The impact of interruption of anomalous systemic arterial supply on scimitar syndrome presenting during infancy
Authors: Uthaman Babu , Abushaban Lulu , Al‐Qbandi Mustafa , Rathinasamy Jebaraj
Published: March 2008

Objectives:

We sought to evaluate the impact of interruption of anomalous systemic arterial supply (ASAS) on clinical course and management outcome of scimitar syndrome (SS) presenting during infancy.

Background:

No systematic study has been reported so far on this subject, although there are conflicting sporadic reports indicating variable effect.

Methods:

Out of 23 children diagnosed to have SS during the past 25 years, 16 symptomatic infants had ASAS. After interrupting ASAS (coil embolization ‐14, surgical ligation ‐2), they were prospectively followed up to define their clinical course and management outcome.

Results:

All 16 infants had sizable ASAS and 9 had variable scimitar vein (SV) stenosis. Fifteen (94%) had pulmonary hypertension and significant left to right shunt. Post intervention, there was variable reduction of shunt in 14 and pulmonary artery pressure in 15 cases. All showed varying clinical improvement. One died of septicemia shortly afterwards. Definitive surgery was deferred for optimal results in seven children for a mean period of 8 months (range 1 month to 3 years). Remaining eight children did not require definitive surgery. One among them had closure of stenosed partial SV by Amplatzer duct occluder (“physiologic correction”). Two children on short term and six children on long term follow‐up (mean 5.2 years; range 3.3–10.3 years) are doing well.

Conclusion:

Interruption of ASAS helps to avoid or defer definitive surgery for SS during infancy. Therefore, we recommend coil embolization of ASAS as initial palliation, and long term surveillance to assess need for further intervention. © 2008 Wiley‐Liss, Inc.
Assessment of operability of congenital cardiac shunts with increased pulmonary vascular resistance
Authors: Viswanathan Sangeetha , Kumar R. Krishna
Published: March 2008

Patients with large left to right shunts as a result of congenital heart disease can develop changes in pulmonary vasculature that are initially reversible. It is critical in these patients to determine whether closure of the defect would reverse some of the changes in the pulmonary vasculature. A comprehensive clinical and noninvasive evaluation often allows classification in the extremes of the spectrum, but for borderline situations, cardiac catheterization is traditionally undertaken. It is important to obtain invasive data meticulously and efficiently recognizing that the numbers only offer a snapshot and may not be representative of the usual physiologic state of the patient. There are, in addition, several caveats that need to be considered while calculating flows and resistances in these patients. Currently a holistic approach that combines clinical, noninvasive, and invasive data may be the only realistic way of making a decision regarding operability in this challenging group of patients with shunt lesions and elevated pulmonary vascular resistance. © 2008 Wiley‐Liss, Inc.
Percutaneous implantation of the first repositionable aortic valve prosthesis in a patient with severe aortic stenosis
Authors: Buellesfeld Lutz , Gerckens Ulrich , Grube Eberhard
Published: March 2008

Objectives and Background:

Percutaneous aortic valve replacement is a new less‐invasive alternative for high‐risk surgical candidates with aortic stenosis. However, the clinical experience is still limited, and the currently available ‘first‐generation devices’ revealed technical shortcomings, such as lack of repositionability and presence of paravalvular leakages. We report the first‐in‐man experience with the new self‐expanding Lotus™ Valve prosthesis composed of a nitinol frame with implemented bovine pericardial leaflets which is designed to address these issues, being repositionable and covered by a flexible membrane to seal paravalvular gaps. We implanted this prosthesis in a 93‐year old patient presenting with severe symptomatic aortic stenosis (valve area: 0.6 cm2). Surgical valve replacement had been declined due to comorbidities.

Methods and Results:

We used a retrograde approach for insertion of the 21‐French Lotus catheter loaded with the valve prosthesis via surgical cut‐down to the external iliac artery. Positioning of the valve was guided by transesophageal echo and supra‐aortic angiograms. The prosthesis was successfully inserted and deployed within the calcified native valve. Echocardiography immediately after device deployment showed a significant reduction of the transaortic mean pressure gradient (32 to 9 mmHg; final valve area 1.7 cm2) without evidence of residual aortic regurgitation. The postprocedural clinical status improved from NYHA‐IV to NYHA‐II. These results remained unchanged up to the 3 month follow‐up.

Conclusions:

Successful percutaneous aortic valve replacement can be performed using the new self‐expanding and repositionable Lotus valve for treatment of high‐risk patients with aortic valve stenosis. Further studies are mandatory to assess device safety and efficacy in larger patient populations. © 2008 Wiley‐Liss, Inc.
Carotid artery dissection: Endovascular treatment. Report of 12 patients
Authors: Fava Mario , Meneses Luis , Loyola Soledad , Tevah Jose , Bertoni Hernan , Huete Isidro , Mellado Patricio
Published: March 2008

Objective:

The purpose of this article is to report our experience with endovascular treatment of internal carotid artery (ICA) dissection with the use of stents.

Background:

Carotid Dissection is an important cause of ischemic stroke in young and middle‐aged patients. There are some patients in whom invasive management is recommended.

Methods:

Twelve patients (eight females, four males), mean age of 50 years (range 35–80 years) with ICA dissection, were treated with the endovascular approach during a 24‐month period. Patients included in this study underwent magnetic resonance (MR) and digital subtraction angiography (DSA) for diagnosis. Over this time period, 162 patients with ICA dissections were seen at our institution. Indications for endovascular treatment were: recurrent ischemic events despite adequate anticoagulant treatment in seven cases, contraindication to anticoagulation in four cases, and one case with significant mismatch between diffusion and perfusion weighted MR. Eleven dissections were spontaneous and one was traumatic. The follow‐up was performed clinically with Doppler ultrasound (US) and MR at 6, 12, and 24 months.

Results:

Stent deployment was successful in all cases. Acute symptoms were resolved in 66.7% of patients. No patients deteriorated their neurological status. There were no new clinical events, stent stenosis or occlusion on 24 months follow‐up.

Conclusion:

Our results showed an excellent clinical outcome of the treated patients. This suggests promising results with the use of endovascular treatment in selected patients. © 2008 Wiley‐Liss, Inc.
State of the art: Management of iliac artery aneurysmal disease
Authors: Michael Bacharach J. , Slovut David P.
Published: March 2008

Although relatively uncommon, isolated iliac artery aneurysms are associated with significant risk of rupture and death. Clinical presentation can be confusing and ultrasound or CT imaging is paramount in establishing the diagnosis and anatomical extent of disease. Important considerations prior to intervention include determination of proximal neck, involvement of the internal iliac artery, and status of the contralateral internal iliac artery. Endovascular repair has evolved as the first choice treatment option for patients with anatomically suitable iliac artery aneurysms. In uncommon circumstances when endovascular treatment may result in significant pelvic ischemia or the primary symptoms are related to extrinsic compression of adjacent structures, surgical repair may be the preferred option. © 2008 Wiley‐Liss, Inc.
Optimal treatment of renal artery in‐stent restenosis: Repeat stent placement versus angioplasty alone
Authors: N'Dandu Zola M. , Badawi Ramy A. , White Christopher J. , Grise Mark A. , Reilly John P. , Jenkins J. Stephen , Collins Tyrone J. , Ramee Stephen R.
Published: March 2008

Objective:

We investigated whether repeat renal artery stent placement compared with treatment with balloon angioplasty alone results in better patency in patients presenting with renal artery in‐stent restenosis (ISR).

Background:

Although stent placement for renal artery stenosis has been demonstrated to be superior to balloon angioplasty for “de novo” renal artery lesions, the optimal therapy for ISR remains unclear.

Methods:

Between January 1997 and August 2006, 34 consecutive patients (41 renal arteries) with ISR were treated at the discretion of the operator with balloon angioplasty or repeat stent placement. Quantitative angiography was performed before and immediately after intervention and at follow‐up. Angiographic follow‐up was obtained for clinical indications in 75% of lesions and routine noninvasive follow‐up imaging was obtained in 95% of lesions.

Results:

Repeat renal artery stent placement demonstrated improved patency compared with balloon angioplasty alone with a 58% reduction in recurrent ISR (29.4% vs. 71.4%, P = 0.02) and a 30% reduction in follow‐up diameter stenosis (41% vs. 58.2%, P = 0.03). The repeat stent group also had better secondary patency (P = 0.05) and a greater freedom from repeat ISR (P = 0.01) when compared with balloon angioplasty alone. There was a trend favoring repeat stent placement for cumulative freedom from target vessel revascularization (TVR) (P = 0.08).

Conclusions:

Repeat stent placement appears to result in superior patency compared with balloon angioplasty alone for the treatment of renal ISR. © 2008 Wiley‐Liss, Inc.
Synthesis of Antiproliferative Cephalotaxus Esters and Their Evaluation against Several Human Hematopoietic and Solid Tumor Cell Lines: Uncovering Differential Susceptibilities to Multidrug Resistance
Authors: Eckelbarger Joseph D. , Wilmot Jeremy T. , Epperson Matthew T. , Thakur Chandar S. , Shum David , Antczak Christophe , Tarassishin Leonid , Djaballah Hakim , Gin David Y.
Published: March 2008

Deoxyharringtonine (2), homoharringtonine (3), homodeoxyharringtonine (4), and anhydroharringtonine (5) are reported to be among the most potent members of the antileukemia alkaloids isolated from the Cephalotaxus genus. Convergent syntheses of these four natural products are described, each involving novel synthetic methods and strategies. These syntheses enabled evaluation of several advanced natural and non‐natural compounds against an array of human hematopoietic and solid tumor cells. Potent cytotoxicity was observed in several cell lines previously not challenged with these alkaloids. Variations in the structure of the ester chain within this family of alkaloids confer differing activity profiles against vincristine‐resistant HL‐60/RV+, signalling new avenues for molecular design of these natural products to combat multi‐drug resistance.
Bcl‐xL antisense oligonucleotide and cisplatin combination therapy extends survival in SCID mice with established mesothelioma xenografts
Authors: Littlejohn James E. , Cao Xiaobo , Miller Steven D. , Ozvaran Mustafa K. , Jupiter Daniel , Zhang Lidong , Rodarte Charles , Smythe W. Roy
Published: March 2008

Bcl‐xL functions as a dominant regulator of apoptotic cell death and is implicated in chemotherapeutic resistance of malignant pleural mesothelioma (MPM). Mesothelioma cell lines demonstrate increasing levels of Bcl‐xL as resistant clones are selected invitro. Moreover, upon introduction of antisense oligonucleotides specific to Bcl‐xL mRNA, MPM cells are sensitized to chemotherapeutic agents. Here we describe the therapeutic effects of a novel combination therapy, Bcl‐xL antisense oligonucleotide (ASO 15999) and cisplatin, on mesothelioma cell lines in vitro and invivo; in addition, efficacy of ASO 15999 in decreasing tumor load as well as its effect on survival in an animal model. Finally, we initiated preliminary toxicity studies involved with intraperitoneal (IP) injections of ASO 15999 into mice. This novel combination, with doses of cisplatin four times below established IC50 levels, significantly decreased viability of MPM cell lines after 48 hr. The growth of established mouse flank human tumor xenografts was reduced with intra‐tumor administration of ASO 15999. Local spread and development of IP xenografts was reduced with treatments of ASO alone, and survival of mice afflicted with these xenografts was prolonged after administration of ASO alone and ASO 15999 + cisplatin combination therapy. These findings suggest that ASO 15999 sensitizes MPM cell lines to the toxic effects of cisplatin. ASO 15999 induced reduction of Bcl‐xL is effective in slowing the progression of human mesothelioma cell lines both in vitro and in vivo. More notably, the combination of Bcl‐xL ASO and cisplatin extends survival in an orthotopic tumor xenograft model. © 2008 Wiley‐Liss, Inc.
How to explore timing of intervention for students at risk of departure
Authors: Ishitani Terry T.
Published: March 2008

Many existing studies have failed to address issues related to the timing of student departure from college. This chapter proposes event history modeling to assess when students depart.
Biomechanical and immunohistochemical properties of meniscal cartilage after high hydrostatic pressure treatment
Authors: Naal Florian D. , Schauwecker Johannes , Steinhauser Erwin , Milz Stefan , von Knoch Fabian , Mittelmeier Wolfram , Diehl Peter
Published: March 2008

Meniscal allograft processing procedures, in particular gamma irradiation, deteriorate the biomechanical and biological properties of the transplanted tissue. High hydrostatic pressure (HHP) treatment, widely used in food technology to inactivate microorganisms while preserving natural compounds, might serve as a gentle alternative to gamma irradiation in the processing of meniscal allografts. We therefore investigated the effects of HHP treatment on the biomechanical and immunohistochemical properties of meniscal cartilage. Specimens of bovine menisci were treated with HHP for 10 min (20°C) at 300 MPa and 600 MPa. Untreated control samples were left at room temperature and ambient pressure. We performed repetitive cycling indentation‐tests to assess the biomechanical properties—in particular the viscoelastic behavior—of HHP treated and untreated meniscal specimens. Immunohistochemical analysis for collagens type I, II, and III and for the proteoglycans versican, aggrecan and for link‐protein was performed by immunolabeling cross‐sections of untreated and at 600 MPa HHP treated specimens. Comparing untreated and HHP treated meniscal specimens there were no significant differences for all tested biomechanical parameters. All cross‐sections of untreated and HHP treated specimens stained positive for the collagens and proteoglycans. We demonstrated that meniscal cartilage can be treated by HHP at levels as high as 600 MPa without affection of the biomechanical and immunochistochemical properties. Therefore, HHP treatment might serve as a gentle alternative to gamma irradiation in the processing of meniscal allografts. Further research is necessary to verificate the present results in vivo. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008
Plasma surface modification of poly(D,L‐lactic acid) as a tool to enhance protein adsorption and the attachment of different cell types
Authors: Alves C. M. , Yang Y. , Marton D. , Carnes D. L. , Ong J. L. , Sylvia V. L. , Dean D. D. , Reis R. L. , Agrawal C. M.
Published: March 2008

We have studied the influence of oxygen radio frequency glow discharge (RfGD) on the surface and bulk properties of poly(D,L‐lactic acid) (PDLLA) and the effect of this surface modification on both protein adsorption and bone cell behavior. PDLLA films were characterized before and after plasma surface modification by water contact angle, surface energy, and adhesion tension of water as well as by scanning electron microscopy (SEM), X‐ray electron spectroscopy (XPS), and Fourier transform infra‐red (FTIR) spectroscopy. RfGD‐films showed an increase in hydrophilicity and surface energy when compared with untreated films. Surface morphological changes were observed by SEM. Chemical analysis indicated significant differences in both atomic percentages and oxygen functional group. Protein adsorption was evaluated by combining solute depletion and spectroscopic techniques. Bovine serum albumin (BSA), fibronectin (FN), vitronectin (VN), and fetal bovine serum (FBS) were used in this study. RfGD‐treated surfaces adsorbed more BSA and FN from single specie solutions than FBS that is a more complex, multi‐specie solution. MG63 osteoblast‐like cells and primary cultures of fetal rat calvarial (FRC) cells were used to assess both the effect of RfGD treatment and protein adsorption on cell attachment and proliferation. In the absence of preadsorbed proteins, cells could not distinguish between treated and untreated surfaces, with the exception of MG63 cells cultured for longer periods of time. In contrast, the adsorption of proteins increased the cells' preference for treated surfaces, thus indicating a crucial role for adsorbed proteins in mediating the response of osteogenic cells to the RfGD‐treated PDLLA surface. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008
Occult hepatitis C virus infection during an outbreak in a hemodialysis unit in Thailand
Authors: Thongsawat Satawat , Maneekarn Niwat , Kuniholm Mark H. , Pantip Chansom , Thungsuputi Amornrat , Lumlertkul Dusit , Bannachak Derek , Nelson Kenrad E.
Published: March 2008

Control of hepatitis C virus (HCV) in hemodialysis populations is a major public health priority, but the preferred methods to prevent and rapidly detect HCV outbreaks in these populations remains subject to debate. We enrolled 231 hemodialysis patients at three dialysis centers in Chiang Mai, Thailand. Patients were followed every 6 months for 3 years and tested for the presence of serum HCV antibody and HCV RNA at each visit. We additionally isolated and tested peripheral blood mononuclear cells (PBMCs) for HCV RNA collected at the 30‐month follow‐up visit. Fifty‐one study participants negative for anti‐HCV at the baseline enrollment visit seroconverted over the course of the 3‐year follow‐up period. Of 11 individuals who transiently lost detectable serum HCV viremia, we were able to detect HCV RNA from the PBMCs of two individuals. Our results suggest that occult HCV infection may be common among hemodialysis patients, and serum HCV RNA testing may be supplemented with PBMC testing to maximize diagnostic sensitivity and aid in outbreak containment. Further work on the diagnostic implications of HCV compartmentalization in hemodialysis and other settings is urgently needed. J. Med. Virol. 80:808–815, 2008. © 2008 Wiley‐Liss, Inc.
Detection of a broad range of human adenoviruses in respiratory tract samples using a sensitive multiplex real‐time PCR assay
Authors: Wong Sallene , Pabbaraju Kanti , Pang Xiaoli L. , Lee Bonita E. , Fox Julie D.
Published: March 2008

Human adenoviruses (hAdVs) are associated with acute respiratory tract infections in pediatric populations and have been identified as a cause of outbreaks in institutional settings. Rapid diagnosis of hAdV infection is critical for appropriate and timely management. This study reports the design and validation of a sensitive and specific multiplex real‐time PCR for the detection of a broad range of hAdV serotypes in respiratory samples. The assay targets the conserved region of the hAdV hexon gene and utilizes hydrolysis probes for the detection of amplified products. The assay was evaluated using retrospectively (n = 864) and prospectively (n = 11,451) collected samples from November 2005 to July 2006. Seasonality studies and analysis of outbreaks was conducted over a 2‐year period from January 2005 to December 2006 (n = 33,067 samples). The assay gave a hAdV positive rate of 7.1% (n = 811) for specimens tested prospectively and was able to detect a broad range of hAdV serotypes with good sensitivity and specificity. A high rate of co‐infection was noted (21.7%). Adenovirus infections were more prevalent in the young with a median age of 24 months for positive patients. Sequence analysis of hAdV positives showed that serotype 7 was the most prevalent followed by serotypes 2 and 3. Association of hAdVs with respiratory outbreaks was low at 2.3% (6 of 266 outbreaks tested) and no seasonal variation was observed for hAdV infections during the 2‐year study period. This assay can improve the detection of hAdVs in respiratory samples and can be used to provide valuable epidemiological information. J. Med. Virol. 80:856–865, 2008. © 2008 Wiley‐Liss, Inc.
Cross‐protection of hepatitis E virus genotypes 1 and 4 in rhesus macaques
Authors: Huang Weijin , Zhang Huayuan , Harrison Tim J. , Lang Shuhui , Huang Guoyong , Wang Youchun
Published: March 2008

The purpose of this study was to determine cross‐protection between HEV genotypes 1 and 4, which are prevalent in China. Fecal suspensions of genotypes 1 and 4 from patients, as well as genotype 4 from swine, were inoculated intravenously into rhesus macaques. Each inoculum contained 5 × 104 genome equivalents of HEV. After infection, serum and fecal samples were collected serially and the levels of alanine aminotransferase (ALT) and anti‐HEV IgG and IgM in sera, and HEV RNA in fecal samples, were measured. Liver biopsies were carried out. All the infected monkeys (12/12) developed anti‐HEV IgG and exhibited fecal shedding of virus. IgM was detected in 11 of 12, and ALT elevation occurred about 2–6 weeks post‐inoculation in 10 of 12, infected monkeys. Hepatic histopathology was consistent with acute viral hepatitis and the ORF2 antigen of HEV was detected in the granular cytoplasm of hepatocytes by immunohistochemistry. After recovery from their initial HEV infection, the monkeys were challenged with a heterologous genotype or heterologous source of HEV and monitored for hepatitis and fecal shedding. Previous infection with HEV completely or partially protected against subsequent challenge with a heterologous virus, because 7 of 11 monkeys did not develop HEV infection or shed virus in the feces, and none of them developed hepatitis or exhibited ALT elevation or liver biopsy findings of hepatitis. In conclusion, previous HEV infection may give rise to cross‐genotype and cross‐host‐species protection. J. Med. Virol. 80:824–832, 2008. © 2008 Wiley‐Liss, Inc.
Dual infection with hepatitis A and E viruses in outbreaks and in sporadic clinical cases: Cuba 1998–2003
Authors: Rodríguez Lay Licel de los Angeles , Quintana Ariel , Villalba María Caridad Montalvo , Lemos Gilda , Corredor Marité Bello , Moreno Aidonis Gutiérrez , Prieto Pablo Aguiar , Guzmán María G. , Anderson David
Published: March 2008

Viral hepatitis ranks as the fifth cause of morbidity for infectious diseases in Cuba. Epidemics are observed frequently in the population, the hepatitis A virus being the main agent responsible for such epidemics. Previous reports also confirmed the circulation of the hepatitis E virus. From 1998 to 2003, 258 serum samples were collected by the Reference Laboratory on Viral Hepatitis during 33 outbreaks of acute viral hepatitis as well as from 39 sporadic clinical cases. Sera were tested for anti‐HAV and anti‐HEV IgM by EIA. Overall of the 33 outbreaks studied sera from 12 (36.4%) were positive for anti‐HAV IgM only, from 7 (21.2%) were positive for anti‐HEV IgM only, and from 14 (42.4%) were positive for antibodies to both viruses. Individually of the 258 sera collected, 137 (53.1%) were positives for anti‐HAV IgM, 20 (7.8%) were positives for anti‐HEV IgM, 33 (12.8%) were positives for both markers and 68 (26.4%) were negative to both. Of the clinical cases, 4 (10.3%) were positives for anti‐HAV IgM, 13 (33.3%) were positives for anti‐HEV IgM and 5 (12.8%) were positives for both markers. Seventeen (43.6%) sera were negatives for all viral hepatitis markers available (A–E). A high positivity for HEV was found in outbreaks tested with the kit produced by CIGB. In particular HEV seems to infect individuals of all ages. The results demonstrate the co‐circulation of and co‐infection with two enterically transmitted viruses; however a higher positivity was observed for anti‐HAV than to anti‐HEV (53.1% vs. 7.8%) in outbreaks. J. Med. Virol. 80:798–802, 2008. © 2008 Wiley‐Liss, Inc.
Recombinant norovirus implicated in gastroenteritis outbreaks in Hiroshima prefecture, Japan
Authors: Fukuda Shinji , Sasaki Yukie , Takao Shinichi , Seno Masato
Published: March 2008

Norovirus (NoV) is a major etiological agent of acute gastroenteritis outbreaks worldwide. A total of 314 fecal specimens collected from patients of 39 NoV gastroenteritis outbreaks in Hiroshima Prefecture, Japan, between December 2001 and April 2006 were tested for the occurrence of recombinant NoVs. Sixteen genotypes (GI/1, GI/2, GI/4, GI/7, GI/8, GI/11, GI/14, GII/2, GII/3, GII/4, GII/5, GII/6, GII/8, GII/12, GII/14, and GII/untypeable) were detected in the 39 outbreaks based on capsid sequences and GII/4 was predominant recently. Twelve strains detected in 11 (28.2%) of the 39 outbreaks were suspected to be recombinants by using Simplot and Recco analyses and five recombinant genotypes, GII/4‐GII/12 (five strains), GIIb‐GII/3 (four strains), GII/4‐GII/2 (one strain), GII/4‐GII/14 (one strain), and GI/2‐GI/8 (one strain), were identified based on RNA‐dependent RNA polymerase and capsid sequences. None of the strains genotyped as GII/4 based on the capsid sequence was identified as a recombinant. The putative recombination points in the recombinant strains were placed either upstream or downstream of the open reading frame (ORF) 1 and ORF2 overlap. The present study indicates the following: (a) recombination among ORFs is common in nature, (b) the involvement of recombinant NoVs in gastroenteritis outbreaks is extensive even in a local area such as Hiroshima Prefecture, Japan, and (c) the conserved region (ORF1 and ORF2 overlap) has a meaningful function against the recombination event. J. Med. Virol. 80:921–928, 2008. © 2008 Wiley‐Liss, Inc.
Expression levels of MDR1, MRP1, MRP4, and MRP5 in peripheral blood mononuclear cells from HIV infected patients failing antiretroviral therapy
Authors: Turriziani Ombretta , Gianotti Nicola , Falasca Francesca , Boni Arianna , Vestri Anna Rita , Zoccoli Alice , Lazzarin Adriano , Antonelli Guido
Published: March 2008

The aim of the study was to evaluate the mRNA expression of four relevant ABC‐transporter genes [MDR1 (P‐glycoprotein; Pgp), MRP1, MRP4, and MRP5] in HIV‐positive individuals failing treatment and analyze the association between the levels of their expression and viral load, CD4 cell count, and therapeutic history. Ninety‐eight HIV‐positive samples and 20 samples from healthy donors were analyzed, retrospectively. Peripheral blood mononuclear cells (PBMCs) from HIV1‐positive individuals were collected at the time of virological failure. Expression of mRNA of Pgp, MRP1, MRP4, and MRP5 in PBMCs was evaluated by real‐time PCR. A high inter‐individual variability was observed in both HIV‐positive individuals and healthy donors but the expression levels of all mRNA analyzed were significantly higher in the HIV‐infected group (P < 0.05). A weak but significant inverse correlation was observed between CD4 cell counts and expression levels of MRP4 and MRP5. Comparison of mRNA expression between individuals with different therapeutic histories showed that expression of MRP4 and MRP5 genes in patients who were both protease inhibitor (PI) and non‐nucleoside reverse transcriptase inhibitor (NNRTI)‐experienced was significantly higher than in patients who were PI experienced but NNRTI‐naïve. In conclusion, the mRNA expression of Pgp, MRP1, MRP4, and MRP5 varies among HIV‐infected patients and healthy donors but is significantly higher in HIV‐positive patients than in donors. The expression of MRP4 and MRP5 seems to correlate with CD4 cell counts. The same protein seems to be overexpressed in patients receiving NNRTIs. J. Med. Virol. 80:766–771, 2008. © 2008 Wiley‐Liss, Inc.
Structural effects of amino acid variations between B and CRF02‐AG HIV‐1 integrases
Authors: Malet Isabelle , Soulie Cathia , Tchertanov Luba , Derache Anne , Amellal Bahia , Traore Ousmane , Simon Anne , Katlama Christine , Mouscadet Jean‐François , Calvez Vincent , Marcelin Anne‐Geneviève
Published: March 2008

HIV‐1 integrase is one of the three essential enzyme required for viral replication and has a great potential as a novel target for anti‐HIV drugs. The sequence variability of the entire integrase (IN) was examined in HIV‐1 subtype B and CRF02‐AG antiretroviral naïve infected patients for the presence of naturally occurring polymorphisms IN gene sequences and protein structures from both subtypes were compared. The phylogenetic analysis showed a total concordance between the 3 pol gene sequences for patients identified as subtype B whereas 3% of patients identified as CRF02‐AG showed a mixture of subtypes. The analysis of IN aa sequences showed that 13 positions (K/R14, V/I31, L/I101, T/V112, T/A124, T/A125, G/N134, I/V135, K/T136, V/I201, T/S206, L/I234, and S/G283) differed between subtypes B and CRF02‐AG. As observed in the 3D model of the preintegration complex, these differences may impact the functional property of IN. The fact that most variations were grouped suggests that some of them are linked together through compensatory mechanisms. This comparison allowed us to identify several variations of amino acids in HIV‐1 IN subtype CRF02‐AG that could have a putative impact on anti‐integrase sensitivity. In particular, the region formed by Thr125, Thr124, Val31 contains at least one residue, T125, which variation has been involved in eliciting resistance to the naphtyridine carboxamide L870,810 IN inhibitor. In conclusion, virological response to anti‐integrase should be studied carefully, according to the subtype, in clinical trials. J. Med. Virol. 80:754–761, 2008. © 2008 Wiley‐Liss, Inc.
Noninvasive blood flow imaging for real‐time feedback during laser therapy of port wine stain birthmarks
Authors: Huang Yu‐Chih , Ringold Tyson L. , Nelson J. Stuart , Choi Bernard
Published: March 2008

Background and Objectives

During laser therapy of port wine stain (PWS) birthmarks, regions of persistent perfusion may exist. Immediate retreatment of such regions may improve PWS laser therapeutic outcome. To address this need, we propose use of laser speckle imaging (LSI) to provide real‐time, quantitative feedback during laser surgery. Herein, we present in vitro and in vivo data collected with a clinic‐based LSI instrument.

Study Design/Materials and Methods

Prior to clinical implementation, we first investigated three aspects of LSI deemed important for clinical imaging: (1) instrument depth of field (DOF); (2) effects of laser irradiance on speckle flow index (SFI) values; and (3) measurement repeatability. Clinical measurements were acquired from the lesions of PWS patients immediately prior to and after laser therapy at the Beckman Laser Institute.

Results

Our preclinical data suggest the following: (1) instrument DOF was ∼1 cm; (2) quantitative flow characterization with LSI was practically unaffected at normalized irradiance values between 0.06 and 0.5; and (3) our LSI instrument was capable of highly reproducible SFI values. From our clinical measurements, we found that the relative difference between blood perfusion in PWS lesions and adjacent normal skin was highly variable. Based on SFI images, the perfusion of PWS skin is sometimes indistinguishable from that of adjacent normal skin. With laser therapy, we measured a global decrease in blood perfusion, and we frequently observed distinct regions of persistent perfusion.

Conclusions

Our results demonstrate the potential role of image‐guided laser therapy of PWS birthmarks. LSI is a promising tool for noninvasive blood flow characterization during laser therapy due to its relative simplicity and low cost. Laser Surg. Med. 40:167–173, 2008. © 2008 Wiley‐Liss, Inc.