bluuebunny/crossref_metadata_2025 huggingface dataset:
Acellular Dermal Allograft for Sellar Reconstruction after Transsphenoidal Hypophysectomy
Authors: Citardi Martin J. , Cox Artemus J. , Bucholz Richard D.
Published: August 2007

Obliteration of the sphenoid sinus using fat is often used after transsphenoidal hypophysectomy. The morbidity of this approach includes donor site complications, fat necrosis, and delayed mucocele formation. As obliteration with fat is intended to prevent cerebrospinal fluid (CSF) leakage, an alternative for this technique would be techniques used for CSF rhinorrhea repair. Instead of sinus obliteration, these defects are repaired with fascial autografts, which are unfortunately associated with donor site complications. To avoid sinus obliteration and donor site complications, we have reconstructed the sella with acellular dermal allograft in lieu of sinus obliteration. Transsphenoidal hypophysectomy was performed under combined microscopic and endoscopic visualization. For closure, the sellar anterior wall was reconstructed with acellular dermal allograft, septal cartilage/bone autograft, and fibrin glue. The sinus mucosa was then draped over the reconstruction and held in place with microfibrillar collagen hemostat slurry. The sphenoid sinus was not obliterated. Postoperatively, all patients underwent serial nasal endoscopy. Thirteen patients underwent the procedure as described for removal of pituitary adenoma. Postoperative discomfort and pain were minimal. Intraoperative CSF leaks were identified in five patients; none of these patients experienced a postoperative CSF leak. The microfibrillar collagen hemostat was cleared by sphenoid mucociliary clearance. One patient developed acute sphenoid sinusitis several weeks after surgery; this patient did not develop meningitis. One postoperative CSF leak occurred in an obese patient, in whom an intraoperative CSF leak was not identified; this leak resolved with bedrest and delayed lumbar drainage alone. Sellar reconstruction with acellular dermal allograft may eliminate the need for sphenoid sinus obliteration after transsphenoidal hypophysectomy. Acellular dermal allograft sellar reconstruction ultimately provides for an aerated, functioning sphenoid sinus without increased CSF leak risk or potential donor site morbidity.
Osteoplastic Flap versus Modified Endoscopic Lothrop Procedure in Patients with Frontal Sinus Disease
Authors: Ulualp Seckin O. , Carlson Thomas K. , Toohill Robert J.
Published: August 2007

The aim of the present study was to review the clinical results of osteoplastic flap procedure with abdominal fat obliteration and modified endoscopic Lothrop procedure. Charts of patients with frontal sinus disease who underwent osteoplastic flap procedure with abdominal fat obliteration or modified endoscopic Lothrop procedure were retrospectively reviewed. Forty-three patients with frontal sinus disease underwent osteoplastic flap procedure with abdominal fat obliteration. Frontal sinus disease was chronic sinusitis in 21, mucocele in 18, and papilloma in four. None of the patients had recurrence within 3 to 12 years follow-up period. Six patients had decreased forehead sensation, one had a CSF leak, and one had toss of the fat graft. Fifteen patients with chronic frontal sinusitis underwent modified endoscopic Lothrop procedure. The follow-up period ranged from 0.5 to 2.5 years. Two patients had recurrence of disease 2 and 6 months after surgery and required osteoplastic flap. In patients with chronic frontal sinusitis, both procedures achieved good relief of symptoms; however, follow-up time of modified endoscopic Lothrop procedure was smaller than that of osteoplastic flap procedure. In conclusion, osteoplastic flap procedure with abdominal fat obliteration provides successful treatment in patients with frontal chronic sinusitis, mucocele, or papilloma. Modified endoscopic Lothrop procedure achieves the relief of symptoms in patients with chronic frontal sinusitis. With the future availability of long term follow-up results, modified endoscopic Lothrop procedure may reduce the number of osteoplastic flap procedures in patients with chronic frontal sinusitis.
Preoperative Sagittal CT Evaluation of the Frontal Recess
Authors: Jacobs Joseph B. , Lebowitz Richard A. , Sorin Alexander , Hariri Suhail , Holliday Roy
Published: August 2007

Endoscopic surgical approaches for chronic frontal sinusitis require the reestablishment of adequate frontal sinus ventilation and drainage for relief of symptoms. After the resection of anterior ethmoid mucosal disease and cellular structure, the anterior to posterior depth of the nasofrontal beak to the base of skull at the insertion of the ethmoidal bulla (frontal sinus ostium) often represents a critical margin for functional success. However, little information concerning this dimension is available. Depending on intraoperative surgical judgment of this distance, extended endoscopic surgical procedures involving additional bone resection may be indicated. These approaches may be hazardous due to the proximity of the cranial cavity and orbit. In addition, secondary stenosis can result from the subsequent inflammatory response. Improved CT imaging, high resolution sagittal reformatting, and computer workstations provide the ability to obtain direct preoperative measurements of the frontal recess. We used a paramedian sagittal section and recorded the maximal anterior to posterior depth from the nasofrontal beak to the base of skull at the insertion of the ethmoidal bulla in 20 patients, 31 sides, undergoing primary endoscopic frontoethmoidectomy. In addition, we found a positive correlation between this distance and agger nasi air cell size measured in the same 31 sides.
Electron Microscopic and Functional Aspects of the Human Vomeronasal Organ
Authors: Jahnke Volker , Merker Hans-Joachim
Published: August 2007

The vomeronasal organ or Jacobson's organ is essential for pheromone detection and reproductive behavior in most mammals. The purpose of this article is to describe the fine structure of the adult human vomeronasal organ in 14 specimens and to discuss functional aspects. Our studies show a duct-like invagination of the epithelium, surrounded by numerous exocrine glands with short ducts; their fine structure suggests serous secretion. In the depth of the invagination, pseudostratified columnar epithelial cells are seen, with plump processes, kinocilia, and microvilli at the apical cell membrane. There are several cell types that differ regarding their organelles and electron density; the light sensory cells exhibit neurofilaments. Underneath the typical basement membrane, in the very vascular lamina propria, numerous myelinated and unmyelinated axons are present. These morphologic findings, which are unique in the human body, suggest that a chemosensory epithelium corresponding to a vomeronasal organ may exist. Its central connections and the possible functional significance for pheromone detection are unknown. Preservation of the vomeronasal organ in endonasal surgery could become important both clinically and medicolegally, should function be demonstrated in humans.
Maxillary Sinus Involution after Endoscopic Sinus Surgery in a Child: A Case Report
Authors: Levine Steven B. , Mitra Sanchayeeta
Published: August 2007

Studies in animal models have suggested that functional endoscopic sinus surgery may affect facial skeletal growth in children, although reviews of large clinical series do not support this observation. This is a case report of a 12 year old male referred to the senior author (SBL) several months after undergoing bilateral functional endoscopic sinue surgery. The preoperative computed tomograms of the paranasal sinuses were normal, with symmetrical well-developed paranasal sinuses. Postoperative computed tomography revealed nearly total involution of the osseous skeleton of the left maxillary sinus. This is the first clinical report of alterations in the facial skeleton of a child secondary to iatrogenic trauma directed at the osteomeatal complex. The case and related literature are reviewed in detail.
Nontraumatic Nasal Septal Abscesses in the Immunocompromised: Etiology, Recognition, Treatment, and Sequelae
Authors: Shah Saurabh B. , Murr Andrew H. , Lee Kelvin C.
Published: August 2007

Proper management of a nasal septal abscess requires prompt diagnosis, adequate surgical drainage, and antibiotics to prevent the potentially dangerous spread of infection and the development of severe functional and cosmetic sequelae. Most septal abscesses are the result of trauma to the nose with septal hematoma and subsequent infection. We present our experience with nasal septal abscesses in five immunocompromised patients without history of nasal trauma. All patients were treated with surgical drainage and antibiotics. The infections in four patients resolved, whereas in the fifth, the infection led to death. We report these cases to depict alternate etiologies of nasal septal abcess, particularly in the immunocompromised patient. Our review illustrates the wide spectrum of disease presentation, provides treatment strategies, and emphasizes the potentially catastrophic sequelae of this disease when unrecognized. With the growing number of immunocompromised individuals, it is important to recognize the potential for immunocompromise to influence the development of septal abscess.
Marine Natural Products. Key Advances to the Practical Application of this Resource in Drug Development
Authors: Hamann Mark T. , Hill Russell , Roggo Silvio
Published: August 2007

Over a half century has passed since the first discovery of spongothymidine and spongouridine. These sponge-derived natural products initiated a chain of events that has resulted in countless lives saved from viral infections and cancer. In addition these humble marine products inspired
the critical evaluation of marine life for the production of novel chemistry and new drug leads. The resulting natural products chemistry from these efforts is unprecedented in regard to structural complexity and biological activity. The drug leads based on marine natural products have however
created unique challenges in scaleable production and structural optimization to evaluate toxicity and enhance biological activity. In this report we have focused our discussion on the progress toward the development of general methodologies for the production, optimization and development
of rare but promising marine natural product drug leads. Key developments discussed briefly include sourcing, lead optimization, molecular biology and phylogeny, marine microbial culture, and tools for structure assignment.
Burning the Hay to Find the Needle – Data Mining Strategies in Natural Product Dereplication
Authors: Wolf D. , Siems K.
Published: August 2007

The acquisition and use of data from the LC/MS-ELSD analysis of extracts is described. The methodology requires MS spectra to be recorded in the positive/negative ESI mode, as well as the determination of retention time and peak area from ELSD. Subsequent calculation of molecular weight,
referenced retention time, and normalized peak area, results in the creation of a peak library, which can be used for different data mining strategies: i) the dereplication of previously isolated natural products; ii) clustering/ranking of extracts for the creation of highly diverse natural
product libraries; iii) a selection tool for the focused isolation of bioactive natural products and iv) to search for alternative sources of a target natural product.
Pharmacophore-Based Screening for the Successful Identification of Bio-Active Natural Products
Authors: Langer Thierry , Laggner Christian , Rollinger Judith M. , Stuppner Hermann
Published: August 2007

We report on the impact of pharmacophore-based virtual screening in the field of drug discovery from natural products. Confronted by an increasing number of secondary metabolites and an increasing number of biomolecular targets relevant in the therapy of human disorders, there is clearly
the need for efficient data management. Filtering of compounds by virtual screening experiments already showed great effect when dealing with large libraries of potential bioactive molecules. Feature-based 3D pharmacophores have been successfully utilized for database mining in order to retrieve
potentially bio-active molecules. However, for the discovery of natural lead candidates, the application of such in silico tools has been so far almost neglected. There seems to be several reasons for this. One concerns the scarce availability of natural product (NP) 3D databases in
contrast to synthetic/combinatorial compound libraries; another reason might be the problematic compatibility of NPs with modern robotized high-throughput-screening (HTS) technologies. Additionally, the incalculable availability of pure natural compounds and their often too complex chemistry
is claimed to hamper such an approach. Thus, research in this field appears time-consuming, highly complex, expensive, and ineffective. Nevertheless, naturally derived compounds are still among the most favorable sources of new drug candidates. A more rational and economic search for new lead
structures originating from nature must therefore be a priority in order to overcome these problems.
Cheminformatic Analysis of Natural Products and their Chemical Space
Authors: Wetzel Stefan , Schuffenhauer Ansgar , Roggo Silvio , Ertl Peter , Waldmann Herbert
Published: August 2007

Cheminformatic methods allow the detailed characterization of particular and characteristic properties of natural products (NPs) and comparison with related characteristics of drugs and other compounds. An overview of the most important properties of natural products and analogues and
their difference with respect to drugs and synthetic compounds is presented. Moreover, different approaches to charting the chemical space populated by natural products are reviewed and their underlying principles are delineated. Some insights about NP chemical space are described together
with possible applications of methods charting chemical space. Strengths and weaknesses of the different approaches will be discussed with respect to possible applications in compound collection design.
P1,P3 Truncated Analogs of Oscillarin and their Inhibitory Activity against Blood Coagulation Factors
Authors: Hanessiana Stephen , Guillemette Sébastien , Ersmark Karolina
Published: August 2007

Based on modeling and available X-ray co-crystal structure data of oscillarin with the enzyme thrombin, a series of P1,P3 truncated analogs were prepared using the azabicyclic octahydroindole carboxamide core as a scaffold. The P1 subunit of the original
natural product was replaced by a 4-amidinobenzamide group, and the P3 subunit was simulated by N-benzylsulfonyl glycine amide or an N-acetyl D-phenylalanine amide. Single digit micromolar inhibition was found against trypsin, thrombin, Factor Xa, and Factor XIa for some analogs.
UZH Hosts Second Dorothy Crowfoot Hodgkin Symposium and Awards First Hoffmann Centennial Medal
Authors: Finney Nathaniel S.
Published: August 2007

On Friday, April 27th, the Organic Chemistry Institute (OCI) hosted the second Dorothy Crowfoot Hodgkin Symposium at the University of Zurich, as well as the presentation of the Hoffmann Centennial Medal for the Study of Natural Products. Organized by Prof. Kim Baldridge, the annual
DCH symposium commemorates the remarkable scientific and social legacy of Dorothy Crowfoot Hodgkin (See http://www.oci.unizh.ch/diversa/dch/2007/ for further information.)
The How, Whom, and Why of Parents’ Involvement in Children’s Academic Lives: More Is Not Always Better
Authors: Pomerantz Eva M. , Moorman Elizabeth A. , Litwack Scott D.
Published: August 2007

A key goal of much educational policy is to help parents become involved in children’s academic lives. The focus of such efforts, as well as much of the extant research, has generally been on increasing the extent of parents’ involvement. However, factors beyond the extent of parents’ involvement may be of import. In this article, the case is made that consideration of the how, whom, and why of parents’ involvement in children’s academic lives is critical to maximizing its benefits. Evidence is reviewed indicating that how parents become involved determines in large part the success of their involvement. It is argued as well that parents’ involvement may matter more for some children than for others. The issue of why parents should become involved is also considered. Implications for future research and interventions are discussed.
The Role of Nonlinear Pedagogy in Physical Education
Authors: Chow Jia Yi , Davids Keith , Button Chris , Shuttleworth Rick , Renshaw Ian , Araújo Duarte
Published: August 2007

In physical education, the Teaching Games for Understanding (TGfU) pedagogical strategy has attracted significant attention from theoreticians and educators for allowing the development of game education through a tactic-to-skill approach involving the use of modified games. However, some have proposed that as an educational framework, it lacks adequate theoretical grounding from a motor learning perspective to empirically augment its perceived effectiveness. The authors examine the literature base providing the theoretical underpinning for TGfU and explore the potential of a nonlinear pedagogical framework, based on dynamical systems theory, as a suitable explanation for TGfU’s effectiveness in physical education. Nonlinear pedagogy involves manipulating key task constraints on learners to facilitate the emergence of functional movement patterns and decision-making behaviors. The authors explain how interpreting motor learning processes from a nonlinear pedagogical framework can underpin the educational principles of TGfU and provide a theoretical rationale for guiding the implementation of learning progressions in physical education.
Doxorubicin and β-Lapachone Release and Interaction with Micellar Core Materials: Experiment and Modeling
Authors: Sutton Damon , Wang Shihu , Nasongkla Norased , Gao Jinming , Dormidontova Elena E.
Published: August 2007

Polymer micelles with two different core-forming blocks, poly(d,l -lactide) (PLA) and poly(ε-caprolactone) (PCL), but the same coronal material, poly(ethylene glycol) (PEG), were investigated in this study as nanoscopic drug carriers. The release of two different drugs, doxorubicin (DOX) and β-lapachone (β-lap), from PEG(5k)-b-PCL(5k) and PEG(5k)-b-PLA(5k) micelles was studied at pH 5.0 and 7.4. Mathematical solutions of both Higuchi’s model and Fickian diffusion equations were utilized to elucidate the differences between the micelle core materials for the two drugs. The neutral and smaller of the two drugs tested, β-lap, demonstrated faster, pH-independent release, suggesting that no substantial changes occurred in either micelle core at lower pH. In contrast, the release rate of DOX was found to noticeably increase at lower pH with a larger cumulative amount of drug released. Different core materials were shown to have considerable influence on the release kinetics of both drugs: in both cases, the more hydrophobic PCL core showed slower drug release rates compared with the less hydrophobic PLA core.
Measurement of Soluble Epoxide Hydrolase (sEH) Activity
Authors: Morisseau Christophe , Hammock Bruce D.
Published: August 2007

The human soluble epoxide hydrolase (sEH; EC 3.3.3.2) is the product of the EXPH2 gene. The sEH catalyzes the addition of a water molecule to an epoxide, resulting in the corresponding diol. Early work suggested a role of sEH in detoxifying a wide array of xenobiotic epoxides; however, recent findings clearly implicate the sEH in the regulation of blood pressure, pain, and inflammation through the hydrolysis of endogenous epoxy fatty acids such as epoxyeicosatrienoic acids (EETs). Both expression and activity of sEH are influenced by a wide array of xenobiotics, underlying how environmental contaminants could influence human health through sEH. This unit describes radiometric, fluorimetric, and mass spectrometric assays for measuring the activity of sEH and its inhibition. Curr. Protoc. Toxicol. 33:4.23.1‐4.23.18. © 2007 by John Wiley & Sons, Inc.
In Vitro Evaluation of Metabolic Drug‐Drug Interactions: A Descriptive and Critical Commentary
Authors: Li Albert P.
Published: August 2007

Adverse drug‐drug interactions represent a major challenge for the pharmaceutical industry. Recently, in vitro approaches for the evaluation of metabolism‐related drug‐drug interactions have been developed. These in vitro approaches are found to be useful in the assessment of clinical drug‐drug interaction potential of new chemical entities and to aid the understanding of clinically significant drug‐drug interactions observed with existing drugs. The general methods for the evaluation of drug‐drug interactions using in vitro, human‐based experimental systems are described and critically reviewed.Curr. Protoc. Toxicol. 33:4.25.1‐4.25.11. © 2007 by John Wiley & Sons, Inc.
Fluorescence Microscopy and Flow Cytometric Analysis of Peyer's Patches and Intestinal Immune Cells
Authors: López María C.
Published: August 2007

In recent years researchers have become more aware of the importance of the gut‐associated immune system since it is in direct interaction with the entry site for virus, bacteria, and all type of food contaminants, including numerous toxins that can alter mucosal immunity. Peyer's patches (PP) are considered the inductive site for protein antigen presentation in the gut as well as the starting point for IgA B‐cell differentiation. The IgA found in feces comes from IgA secreted by IgA lamina propria lymphocytes (LPL), and its presence is a sign of normal physiology, in that IgA plays a role in absorption and immune defense against gut‐associated pathogens. Methods presented in this unit are intended to analyze PP and intestinal intraepithelial and LPL to determine whether the complexity of the mucosal‐associated lymphoid tissue and its components have been altered by any form of external damage. The protocols explain how to isolate and culture isolated cells; how to stain and analyze; and also how to cryopreserve the gut. Curr. Protoc. Toxicol. 33:18.13.1‐18.13.20. © 2007 by John Wiley & Sons, Inc.
Purification and Characterization of Gingipains
Authors: Potempa Jan , Nguyen Ky‐Anh
Published: August 2007

Gingipains are cysteine proteases produced in large quantities by Porphyromonas gingivalis which together constitute important virulence factors in the pathogenesis of periodontal disease by that organism. Described is this unit is an efficient procedure for the purification of gingipains from the growth medium of P. gingivalis strain HG66, along with detailed protocols for growth of the organism and basic characterization of the purified proteases. The purification procedure consists of acetone precipitation followed by gel filtration to separate high‐molecular‐mass gingipains (Kgp and HRgpA) from RgpB. Kgp and HRgpA are further separated on Arg‐Sepharose by the virtue of differential elution from the affinity matrix with lysine (Kgp) and arginine (HRgpA) eluant. Obtained from these procedures, the gingipains are stable and can be stored at −80°C for years without loss of activity. Curr. Protoc. Protein Sci. 49:21.20.1‐21.20.27. © 2007 by John Wiley & Sons, Inc.
Microplate Assay For Cathepsin Detection in Viable Cells Using Derivatives of 4‐Methoxy‐β‐Naphthylamide
Authors: Ruettger Anke , Wiederanders Bernd
Published: August 2007

This unit describes an assay for the direct and selective detection of the four cathepsins B, H, K, and L in adherently growing cells. Cells are incubated with substrates that are peptidic derivatives of 4‐methoxy‐β‐naphthylamine partially selective for each cathepsin, together with 5‐nitrosalicylaldehyde. The protease reaction is performed in microtiter plates and the fluorescent hydrolysis products are detected using a plate reader. The selectivity of detection is enhanced by parallel assays containing inhibitors that are also partially selective for each of the cathepsins. Individual cathepsin activities can then be calculated by the difference between the uninhibited and the inhibited assays. Detection of cathepsin H activity differs from the other assays in that other nonlysosomal aminopeptidases are inhibited by bestatin. The most common application of these assays is to compare directly cells treated with different substances, e.g., pharmaceutically interesting cathepsin inhibitors. Curr. Protoc. Protein Sci. 49:21.21.1‐21.21.8. © 2007 by John Wiley & Sons, Inc.
Detection of Intracellular Cytokines by Flow Cytometry
Authors: Foster Barbara , Prussin Calman , Liu Fei , Whitmire Jason K. , Whitton J. Lindsay
Published: August 2007

Intracellular cytokine detection by flow cytometry has emerged as the premier technique for studying cytokine production at the single‐cell level. Multiparameter flow cytometry permits simultaneous detection of two or more cytokines within a single cell, allowing direct TH1 versus TH2 determination. This capability, combined with the high throughput inherent in the instrumentation, gives intracellular cytokine staining an enormous advantage over existing single‐cell techniques such as ELISPOT, limiting dilution, and T cell cloning. The unit describes intracellular staining of cells that have already been stimulated in vitro and fixed. Methods for in vitro activation by PMA and ionomycin or antigens, fixation of cell suspensions, and cell‐surface staining are also described. Because of the greater level of nonspecific binding inherent in fixed, permeabilized cells, greater care must be taken in designing specificity controls. A protocol in which brefeldin A is used to identify cytokine‐producing T cells activated in response to in vivo contact with microbial antigens is also described. Curr. Protoc. Immunol. 78:6.24.1‐6.24.21. © 2007 by John Wiley & Sons, Inc.
Measurement of Interleukin‐21
Authors: Zeng Rong , Spolski Rosanne , Leonard Warren J.
Published: August 2007

This unit describes three procedures for measurement of interleukin‐21 (IL‐21). The first employs the use of an antibody sandwich ELISA. An alternative procedure measures proliferative responses of T cells to a combination of IL‐21 and IL‐15 using CFSE. Finally, a method to assess IL‐21‐induced tyrosine phosphorylation of Stat3 in splenic CD8+ T cells using a flow cytometry–based analysis is described. Curr. Protoc. Immunol. 78:6.30.1‐6.30.8. © 2007 by John Wiley & Sons, Inc.
Single‐Cell Phospho‐Protein Analysis by Flow Cytometry
Authors: Schulz Kenneth R. , Danna Erika A. , Krutzik Peter O. , Nolan Garry P.
Published: August 2007

This protocol describes methods for monitoring intracellular phosphorylation‐dependent signaling events on a single‐cell basis. This approach measures cell signaling by treating cells with exogenous stimuli, fixing cells with formaldehyde, permeabilizing with methanol, and then staining with phospho‐specific antibodies. Thus, cell signaling states can be determined as a measure of how cells interact with their environment. This method has applications in clinical research as well as mechanistic studies of basic biology. In clinical research, diagnostic or drug efficacy information can be retrieved by discovering how a disease affects the ability of cells to respond to growth factors. Basic scientists can use this technique to analyze signaling events in cell lines and human or murine primary cells, including rare populations, like B1 cells or stem cells. This technique has broad applications to take standard biochemical analysis into primary cells to garner valuable information about signaling events in physiologic settings. Curr. Protoc. Immunol. 78:8.17.1‐8.17.20. © 2007 by John Wiley & Sons, Inc.
Phenotypic Analysis of Human Embryonic Stem Cells
Authors: Ungrin Mark , O'Connor Michael , Eaves Connie , Zandstra Peter W.
Published: August 2007

Human embryonic stem cells (hESCs) are an important tool for the study of developmental biology and may one day serve as a source of cells for regenerative medicine. As no definitive assay for hESC pluripotency is available, surrogate assays that measure markers or properties that have been correlated with hESC developmental potential are used to measure the effects of test conditions on their propagation and differentiation. This unit presents a range of protocols, including visual inspection, flow cytometry, immunofluorescence, quantitative real‐time reverse‐transcriptase PCR, and a colony‐forming assay, as tools to measure the undifferentiated hESC state. The authors discuss the advantages and limitations of the various protocols, and present expected results and discuss potential problems. The development of quantitative assays of hESC developmental potential are critical for our understanding of hESC biology. Curr. Protoc. Stem Cell Biol. 2:1B.3.1‐1B.3.25. © 2007 by John Wiley & Sons, Inc.
Pancreas Differentiation of Mouse ES Cells
Authors: Micallef Suzanne J. , Li Xueling , Elefanty Andrew G. , Stanley Edouard G.
Published: August 2007

This unit describes the derivation of pancreatic cells from mouse embryonic stem cells (ESCs). Mouse ESCs are pluripotent immortal cells derived from the inner cell mass of pre‐implantation blastocyst‐stage embryos that possess the ability to differentiate into any cell type within the adult animal. In vitro, ESCs can be differentiated into a variety of cell types representing derivatives of the three embryonic germ layers, mesoderm, endoderm, and ectoderm. Successfully differentiating ES cells to pancreatic cells has the potential to provide an alternative to cadaver‐derived cells for treatment of type I diabetes. This unit outlines a method for the differentiation of ESCs toward pancreatic endoderm in serum‐free medium from embryoid bodies (EBs) formed in suspension or spin EBs. In addition there is a protocol for maintaining ESC. Curr. Protoc. Stem Cell Biol. 2:1G.2.1‐1G.2.8. © 2007 by John Wiley & Sons, Inc.
Growing Oral Biofilms in a Constant Depth Film Fermentor (CDFF)
Authors: Pratten Jonathan
Published: August 2007

In order to grow organisms in such a manner as to mimic their physiological growth state in vivo, it is often desirable to grow them as biofilms in the laboratory. There are numerous systems available to accomplish this; however, some are more suited to the growth of oral biofilms (dental plaque) than others. The operating parameters of one such model, the constant depth film fermentor (CDFF), are given in this unit. This model is particularly suited to studying the varied biofilms which exist in the oral cavity because environmental factors such as the substratum, nutrient source, and gas flow can be altered. Curr. Protoc. Microbiol. 6:1B.5.1‐1B.5.18. © 2007 by John Wiley & Sons, Inc.
Analyses of Mycobacterium tuberculosis Proteins
Authors: Larsen Michelle H. , Biermann Karolin , Jacobs, William R.
Published: August 2007

This unit includes protocols for the isolation of proteins from Mycobacterium tuberculosis. Considerations for working with M. tuberculosis at Biosafety Level 3 containment are also discussed. Curr. Protoc. Microbiol. 6:10A.4.1‐10A.4.5. © 2007 by John Wiley & Sons, Inc.
Laboratory Maintenance of Flavobacterium psychrophilum and Flavobacterium columnare
Authors: Cain Kenneth D. , LaFrentz Benjamin R.
Published: August 2007

Bacteria in the genus Flavobacterium are highly heterogeneous and are comprised of pathogenic and non‐pathogenic species. They are found in a wide variety of environments that include fresh and marine water, soil, and ocean sediments. This unit focuses on laboratory maintenance of two important fish pathogens within this genus. Flavobacterium psychrophilum and F. columnare are fastidious, slow‐growing organisms that require specialized medium for optimal growth in the laboratory. Methods are described for the preparation of media, broth and agar culture, cryopreservation, and enumeration of viable cells for these bacteria. In addition, the diseases caused by these pathogens are described along with methods of diagnosis and treatments. This unit serves as a general guide for the cultivation of F. psychrophilum and F. columnare and discusses some of the difficulties that have been encountered for each. Curr. Protoc. Microbiol. 6:13B.1.1‐13B.1.12. © 2007 by John Wiley & Sons, Inc.
Epstein‐Barr Virus (EBV): Infection, Propagation, Quantitation, and Storage
Authors: Lan Ke , Verma Subhash C. , Murakami Masanao , Bajaj Bharat , Robertson Erle S.
Published: August 2007

Epstein‐Barr virus (EBV) was first reported as the etiological agent of Burkitt's lymphoma in 1964. Since then, EBV has also been associated with nasopharyngeal carcinoma, which is highly prevalent in Southeast Asia, as well as infectious mononucleosis, complications of AIDS, and transplant‐related B cell lymphomas. This virus has further been linked with T cell lymphomas and Hodgkin's disease, establishing the concept of a wide spectrum of EBV‐associated malignant disorders. So far, there are a number of EBV‐infected cell lines established that can be induced for production of infectious viral progeny and that facilitate the study of the mechanism of EBV‐related infection, transformation, and oncogenesis. This unit describes procedures for the preparation of EBV virion particles and in vitro infection of cells with EBV. In addition, procedures for quantitation and storage of the virus are provided. Curr. Protoc. Microbiol. 6:14E.2.1‐14E.2.21. © 2007 by John Wiley & Sons, Inc.
Reverse Transcription‐Polymerase Chain Reaction‐Based Detection of Plant Viruses
Authors: Li Ruhui , Hartung John S.
Published: August 2007

A one‐step reverse transcription–polymerase chain reaction (RT‐PCR) is used to detect two cherry flexiviruses, Cherry green ring mottle virus (CGRMV) and Cherry necrotic rusty mottle virus (CNRMV), in Prunus species. This unit presents procedures for collection of plant samples, preparation of total nucleic acids, viral RNA‐rich or total RNA extracts from plant tissues, and subsequent amplification of the viral targets by one‐step RT‐PCR using a pair of consensus primers. The PCR amplicons are visualized by electrophoresis in a 1% agarose gel containing ethidium bromide in TAE buffer and viewed under ultraviolet light. This procedure is rapid, sensitive, reliable, and cost‐effective and is generally useful on a wide variety of plant/virus systems. The use of a semi‐automatic homogenizer for sample preparation and one‐tube RT‐PCR for virus detection makes this approach ideal for screening large numbers of samples. Curr. Protoc. Microbiol. 6:16C.1.1‐16C.1.9. © 2007 by John Wiley & Sons, Inc.
Extracting Viral RNAs from Plant Protoplasts
Authors: Fabian Marc R. , Andrew White K.
Published: August 2007

The analysis of viral RNA is a fundamental aspect of plant RNA virus research. Studies that focus on viral RNAs often involve virus infections of plant protoplasts (see UNITS 16D.1‐16D.4). Protoplast offer the advantage of simultaneous initiation of infections, which allows for superior temporal and quantitative analyses of viral RNAs. The efficient isolation of intact viral RNA is key to any such investigations. This unit describes two basic protocols for extracting viral RNAs from plant protoplasts. An approach for preparing double‐stranded viral RNA from total RNA pools is also provided. The viral RNA prepared by using these techniques can be used for further analyses such as primer extension, reverse transcription–PCR, and northern blotting. Curr. Protoc. Microbiol. 6:16E.1.1‐16E.1.6. © 2007 by John Wiley & Sons, Inc.
Isolation of Human Peripheral Blood Mononuclear Cells (PBMCs)
Authors: Lan Ke , Verma Subhash C. , Murakami Masanao , Bajaj Bharat , Robertson Erle S.
Published: August 2007

In this appendix, several basic methods are described for preparation of primary B lymphocyte. Curr. Protoc. Microbiol. 6:A.4C.1‐A.4C.9. © 2007 by John Wiley & Sons, Inc.
Individual differences in use of external visualisations to perform an internal visualisation task
Authors: Cohen Cheryl A. , Hegarty Mary
Published: August 2007

Thirty participants performed a novel spatial inference task, which required them to imagine and draw the cross section of a three‐dimensional (3‐D) object. While performing the task, participants could interactively control two computer visualisations (animations) of the object. There were large individual differences in how frequently participants used the computer visualisations, which were related to spatial ability. Use of the interactive visualisations was highly predictive of performance on the cross‐section task and mediated the correlation between spatial ability and performance. These findings suggest that interactive computer visualisations can aid performance on spatial inference tasks, but that they do so only for a subset of individuals who can discover how to best use the additional information that they provide. Copyright © 2007 John Wiley & Sons, Ltd.
Learner control, cognitive load and instructional animation
Authors: Hasler Béatrice Susanne , Kersten Bernd , Sweller John
Published: August 2007

In order to investigate the influence of learner‐controlled pacing in educational animation on instructional efficiency, three versions of an audio‐visual computer animation and a narration‐only presentation were used to teach primary school students the determinants of day and night. The animations were either system‐paced using a continuous animation, learner‐paced using discrete segments or learner paced using ‘stop’ and ‘play’ buttons. The two learner‐paced groups showed higher test performance with relatively lower cognitive load compared to the two system‐paced groups, despite the fact that the ‘stop’ and ‘play’ buttons were rarely used. The significant group differences regarding test performance were obtained only for more difficult, high element interactivity questions but not for low element interactivity questions. Copyright © 2007 John Wiley & Sons, Ltd.
Can the cognitive load approach make instructional animations more effective?
Authors: Ayres Paul , Paas Fred
Published: August 2007

The papers in this themed issue have investigated methods to make animations more effective. The purpose of this paper is to discuss each of the seven empirical papers. The discussion focuses on how they dealt with cognitive load during instruction and problem solving. Critical observations are made on each paper and avenues for future research are proposed. From the overall collection of papers a number of key results are identified and used as a basis for recommending principles for developing effective instructional animations. Lastly, a number of design issues are discussed in the context of enhancing future research in this field. Copyright © 2007 John Wiley & Sons, Ltd.
Diversity and relationship between Iranian ethnic groups: Human dopamine transporter gene (DAT1) VNTR genotyping
Authors: Banoei Mohammad Mehdi , Chaleshtori Morteza Hashemzadeh , Sanati Mohammah Hossein , Panahi Mehdi Shafa Sharait , Majidizadeh Tayebeh , Rostami Maryam , Manshadi Massoumeh Dehghan , Golalipour Masoud
Published: August 2007

The 40‐bp VNTR polymorphism in the 3′ untranslated region of the human DAT1 (dopamine transporter 1) was analyzed in the Iranian ethnic groups in order to examine the influence of geographical and linguistic affiliation on the genetic affinities among the Iranian population. A total of 449 subjects belonging to nine ethnic groups from the Iranian population were included in the study. The screening of 898 chromosomes showed five alleles (6, 7, 8, 9, 10, and 11), of which allele 10 revealed the highest frequency in most regions. Allele 8 was predominant in one ethnicity and occurred more frequently in the center of Iran. This study shows that the DAT1 distribution in Iran has a different pattern from those in other studies, which can contribute to an understanding of differentiation and diversity of Iranian ethnic groups. This polymorphism could represent the genetic diversity among the various ethnic groups of Iran. Am. J. Hum. Biol., 2007. © 2007 Wiley‐Liss, Inc.
Fractionation technology for the petroleum refining industry
Authors: Hug Albert , Faessler Peter W. , Kolmetz Karl , Ng Wai Kiong , Watari Kazuo
Published: August 2007

Distillation Research and Development is moving rapidly with the utilization of computational fluid dynamic (CFD) modeling. Previously, a potential fractionation application would first be developed and fabricated. Then many hours of pilot plant testing would be required to finalize the design. The time line for the development of new distillation equipment has been reduced, leading to advances in fractionation equipment.

Current applications of trays, packings, distributors and feed inlets can be optimized utilizing CFD modeling. CFD Modeling has improved the current generation of fractionation equipment. This modeling is also particularly important in maximizing capacity in tower revamps where the diameter of the tower is fixed. This paper will review the current‐generation fractionation equipment and the utilization of the same in the petroleum refining industry. Copyright © 2007 Curtin University of Technology and John Wiley & Sons, Ltd.
Cannabis, Pain, and Sleep: Lessons from Therapeutic Clinical Trials of Sativex®, a Cannabis‐Based Medicine
Authors: Russo Ethan B. , Guy Geoffrey W. , Robson Philip J.
Published: August 2007

Cannabis sativa L. has been utilized for treatment of pain and sleep disorders since ancient times. This review examines modern studies on effects of Δ9‐tetrahydrocannabinol (THC) and cannabidiol (CBD) on sleep. It goes on to report new information on the effects on sleep in the context of medical treatment of neuropathic pain and symptoms of multiple sclerosis, employing standardized oromucosal cannabis‐based medicines containing primarily THC, CBD, or a 1 : 1 combination of the two (Sativex®). Sleep‐laboratory results indicate a mild activating effect of CBD, and slight residual sedation with THC‐predominant extracts. Experience to date with Sativex in numerous Phase I–III studies in 2000 subjects with 1000 patient years of exposure demonstrate marked improvement in subjective sleep parameters in patients with a wide variety of pain conditions including multiple sclerosis, peripheral neuropathic pain, intractable cancer pain, and rheumatoid arthritis, with an acceptable adverse event profile. No tolerance to the benefit of Sativex on pain or sleep, nor need for dosage increases have been noted in safety extension studies of up to four years, wherein 40–50% of subjects attained good or very good sleep quality, a key source of disability in chronic pain syndromes that may contribute to patients' quality of life.
The File Mover: high‐performance data transfer for the grid
Authors: Anglano Cosimo , Canonico Massimo
Published: August 2007

The exploration in many scientific disciplines (e.g. High‐Energy Physics, Climate Modeling, and Life Sciences) involves the production and the analysis of massive data collections, whose archival, retrieval, and analysis require the coordinated usage of high‐capacity computing, network, and storage resources. To obtain satisfactory performance, these applications require the availability of a high‐performance, reliable data transfer mechanisms, able to minimize the data transfer time that often dominates their execution time. In this paper we present the File Mover, an efficient data transfer system specifically tailored to the needs of data‐intensive applications, that exploits the overlay networks paradigm to provide superior performance with respect to conventional file transfer systems. An extensive experimental evaluation, carried out by means of a proof‐of‐concept implementation of the File Mover for a variety of network scenarios, shows the ability of the File Mover to outperform alternative data transfer systems. Copyright © 2007 John Wiley & Sons, Ltd.
Exact BER analysis of M‐ary orthogonal signaling with MRC over Ricean fading channels
Authors: Paris J. F. , Martos‐Naya E. , Fernández‐Plazaola U.
Published: August 2007

A new approach for analyzing the performance of coherent M‐ary orthogonal signaling with maximal ratio combining over Ricean fading channels is presented here. A new series representation for the bit‐error rate is derived, making it possible to compute error rates with guaranteed accuracy. Copyright © 2007 John Wiley & Sons, Ltd.
Functionalized 8 nm Long Aryleneethynylene Molecular Wire with Alkyne Termini
Authors: Zeng Xianshun , Wang Changsheng , Bryce Martin R. , Batsanov Andrei S. , Sirichantaropass Skon , García‐Suárez Víctor M. , Lambert Colin J. , Sage Ian
Published: August 2007

The synthesis of new conjugated aryleneethynylene derivatives of up to ca. 8 nm molecular length (compound 16) with terminal alkyne substituents and 9,9‐dihexylfluorene units in the backbone is described. Key synthetic steps are Pd‐mediated Sonogashira coupling methodology combined with regioselective removal from the terminal alkyne units of 2‐hydroxy‐2‐propyl protecting groups in the presence of trimethylsilyl groups. The structural and electronic properties of 16 were obtained from DFT calculations: the intramolecular terminal C···C′ distance in its relaxed conformation was found to be 7.8 nm. The calculated distribution of HOMO and LUMO orbitals and the strong blue fluorescence observed for 16 (λmax = 420, 443 nm in CHCl3 solution) are consistent with a highly conjugated penta[(9,9‐dihexyl‐2,7‐fluorenylene)ethynylene] structure. Molecule 16 possesses multifunctionality and is of interest for future molecular electronic device applications.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
Seismic strengthening of an under‐designed RC structure with FRP
Authors: Di Ludovico M. , Prota A. , Manfredi G. , Cosenza E.
Published: August 2007

The opportunities provided by the use of fiber‐reinforced polymer (FRP) for the seismic retrofit of existing reinforced concrete (RC) structures were assessed on a full‐scale three‐story framed structure. The structure, designed only for gravity loads, was subjected to a bi‐directional pseudo‐dynamic (PsD) test at peak ground acceleration (PGA) equal to 0.20g at the ELSA Laboratory of the Joint Research Centre.

The seismic deficiencies exhibited by the structure after the test were confirmed by post‐test assessment of structural seismic capacity performed by nonlinear static pushover analysis implemented on the lumped plasticity model of the structure. In order to allow the structure to withstand 0.30g PGA seismic actions, a retrofit using glass fiber‐reinforced polymer (GFRP) laminates was designed. The retrofit design was targeted to achieve a more ductile
and energy dissipating global performance of the structure by increasing the ductility of columns and preventing brittle failure modes. Design assumptions and criteria along with nonlinear static pushover analysis to assess the overall capacity of the FRP‐retrofitted structure are presented and discussed. After the retrofit execution, a new series of PsD tests at both 0.20g and 0.30g PGA level were carried out. Theoretical predictions are compared with the main experimental outcomes to assess the effectiveness of the proposed retrofit technique and validate the adopted design procedures. Copyright © 2007 John Wiley & Sons, Ltd.