bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasmid Purification, Therapeutic Applications
Authors: Barbosa Hélder S. C. , Marcos João Carlos
Published: April 2010

The present high demand for pure pDNA to be used in non‐viral vectors is expected to increase in the forthcoming years with the approval of products that are already in phase III clinical trials. To supply this demand adequate methods for manufacturing pDNA at large scale are being developed. Purification is a fundamental step on the overall pDNA manufacturing and could account for up to 80% of the total operational costs.

This article describes the several methods that are available for the purification of pharmaceutical grade pDNA. Starting from cell lysis, the several options of chromatographic and non‐chromatographic methods are described and the advantages and disadvantages of each one highlighted. The different types of chromatography, anion exchange, size exclusion, hydrophobic interaction and affinity, are compared and the specificity of each one described. A special focus is given to non‐chromatographic methods like, precipitation, membrane processes and liquid extraction that are becoming more important on the design of cost‐effective processes. The role of new materials like monoliths to increase the capacity of chromatography, and specific ligands to increase the selectivity of several separation methods is also referred. The integration of different methods to design industrial processes is described and four different cGMP processes are analyzed.
Biocatalytic Synthesis of β‐lactam Antibiotics
Authors: Deaguero Andria L. , Blum Janna K. , Bommarius Andreas S.
Published: April 2010

The penicillins, cephalosporins, carbapenems, and monobactams, make up the β‐lactam family of antibiotics. There are currently more than 25 antibiotics in this family that are approved by the United States Food and Drug Administration and over 50 companies that manufacture them. Their four‐membered β‐lactam ring, which is the part of the antibiotic that is responsible for their bactericidal capabilities, is sensitive to acids, bases, and heat. Therefore, the original chemical synthesis processes utilized to make these compounds required subzero temperatures, hazardous chemicals for protecting groups, and large volumes of organic solvents, rendering them both environmentally harmful and economically taxing. The original syntheses are increasingly being replaced with biocatalytic syntheses which are carried out at ambient temperatures, in aqueous medium, and without the use of auxiliary chemicals, rendering them environmentally benign and economically advantageous processes. The most important enzymes utilized for these processes are penicillin G acylases, penicillin V acylases, cephalosporin acylases, and expandases.
Bioseparation, Magnetic Particle Adsorbents
Authors: Peuker Urs Alexander , Thomas Owen , Hobley Timothy John , Franzreb Matthias , Berensmeier Sonja , SchÄFer Maria , Hickstein Birgit
Published: April 2010

Downstream processing using magnetic adsorbent particles, so called magnetic beads, is a promising technology for future bioseparation challenges for high‐value as well as for mid‐priced products. Magnetic adsorbent based separation currently applies the same ligand technologies as chromatographic separation. After binding, magnetic forces are used for the separation of protein loaded beads from crude bio suspensions. The concept has been proven in many proof of principle examples but few pilot scale processes, due to two main hurdles: Improved, cheaper magnetic adsorbents are required, as are improved magnetic separators. This review examines where magnetic bead based separations can fit into a downstream process before studying several state of the art synthesis processes for magnetic beads and their magnetic and sorptive properties. An overview of magnetic separator technology is given with special focus on bioprocessing. Examples of the separation and purification of proteins applying magnetic beads to a biosuspension is shown.
Smart Biomaterials and Biosensors: Intelligent Properties of Biomolecules and Cells in Hybrid Devices
Authors: Marx Kenneth A.
Published: April 2010

Intelligent properties represent discipline‐independent definitions for complex behaviors of natural (biomacromolecules, cells, organisms), artificial, (algorithms, devices) and hybrid (biosensors) systems. Intelligent properties can be thought of as subproperties of complex adaptive systems that possess: (i) self‐maintenance, (ii) adaptivity, (iii) information preservation, and (iv) spontaneous increase in complexity. The intelligent subproperties includeself‐assembly, self‐organization, self‐repair, self‐replication, self‐maintenance, redundancy, self‐diagnosis, recognition, learning, adaptivity, information storage, signaling, and evolution, to name a few. The properties of biomacromolecules, their complexes, cells, signaling networks, and organism subsystems are discussed within the context of their intelligent properties. Specific biomacromolecules' intelligent properties (e.g. DNA, bacteriorhodopsin) offer a range of application and hybrid device opportunities in creating nanostructures, nanocomputing, microelectronics, optical memory designs, and biosensors. Important biosensor nucleic acid and protein class examples are presented that have basic research and clinical applications. Sophisticated levels of intelligent properties exist within evolved control networks in cells. Therefore, cells have added value integrated into smart biosensors. Cellular networks evolved to control important cellular functions and specific examples of gene regulatory control elements are described. Some naturally occurring smart biomaterials have evolved intelligent properties that play critical roles in maintaining the homeostasis and integrity of an organism. An example is the fibrin gel, critical for blood clotting and wound repair. Our aim has been to better integrate molecular and cellular biochemistry with other scientific disciplines within the context of their applications, interfaces, and shared properties by providing a more universal property language —the intelligent properties—to enrich our understanding of biomolecular systems.
Large‐Scale Protein Purification, Self‐Cleaving Aggregation Tags
Authors: Ghazi Iraj , Wood David W.
Published: April 2010

In the mid‐1980s, the incorporation of affinity tags into recombinant proteins by means of genetic engineering created a highly efficient tool for protein purification. A recent advance in this area has been the addition of self‐cleaving protein domains to conventional affinity tags, which allows the tags to self‐cleave and thus eliminates the requirement for proteolytic removal of the tag. In practice, the native target is released from the affinity tag after purification by a simple temperature, chemical, and/or pH change. Although this is a significant advance for affinity‐tag methods, some limitations still arise from the equipment and consumable resin costs associated with these procedures. In this contribution, we describe solutions to these limitations through the development of a new generation of self‐associating and aggregating tags. One of these tags uses the association of phasin proteins to polyhydroxybutyrate granules coexpressed in the host cell, producing a system where the host cell also produces the affinity matrix. The other is based on reversibly precipitating elastin‐like polypeptide tags that can be separated from cellular contaminants by simple heating and centrifugation. Both these tags allow protein purification to take place using simple mechanical methods, thus eliminating the need for conventional affinity apparatus or reagents. Self‐cleaving aggregation‐tag technologies may dramatically reduce recombinant protein purification costs.
Freezing, Biopharmaceutical Products
Authors: Lam Philippe , Moore Jamie
Published: April 2010

During the manufacturing process of biopharmaceutical products, freeze/thaw of protein solutions are routine operations, allowing product shelf‐life to be extended and providing flexibility in the scheduling of the final fill and finish steps. This article discusses the physics of freezing and thawing, the multiple stresses that can affect protein stability during these operations and the strategies that can be used to mediate these problems.
Flexibility in the use of requesting gestures in squirrel monkeys (Saimiri sciureus)
Authors: Anderson James R. , Kuroshima Hika , Hattori Yuko , Fujita Kazuo
Published: April 2010

Three squirrel monkeys, trained to make a requesting gesture, were tested in the presence of a human assistant whose visual attention varied across trials. When food was available in one dish and an empty dish was nearby, the monkeys pointed overwhelmingly toward the former, regardless of where the assistant was looking. Looking at the assistant while pointing (“monitoring”) peaked when she looked at them and when she attempted to engage them in joint attention. When only one dish was present, the monkeys refrained from gesturing if it was empty and if no assistant was present. They gestured more when the assistant made eye contact with them. Furthermore, when the assistant's focus of attention switched from the dish or the ceiling to the monkeys, the latter resumed pointing and increased their monitoring of the assistant. This is the first demonstration of such flexible use of an intentionally communicative requesting gesture in New World monkeys. Am. J. Primatol. 72:707–714, 2010. © 2010 Wiley‐Liss, Inc.
Prolonged effects of short‐term anti‐CD20 B cell depletion therapy in murine systemic lupus erythematosus
Authors: Bekar Kai W. , Owen Teresa , Dunn Robert , Ichikawa Travis , Wang Wensheng , Wang Roger , Barnard Jennifer , Brady Sean , Nevarez Sarah , Goldman Bruce I. , Kehry Marilyn , Anolik Jennifer H.
Published: April 2010

Objective

Although B cells are implicated in the pathogenesis of systemic lupus erythematosus, the role of B cell depletion (BCD) as a treatment is controversial, given the variable benefit in human disease. This study was undertaken to test the effects of BCD therapy in a murine lupus model to better understand the mechanisms, heterogeneity, and effects on disease outcomes.

Methods

(NZB × NZW)F1 female mice with varying degrees of disease severity were treated with an anti‐mouse CD20 (anti‐mCD20) antibody (IgG2a), BR3‐Fc fusion protein (for BAFF blockade), or control anti‐human CD20 monoclonal antibody (∼10 mg/kg each). Tissue samples were harvested and analyzed by flow cytometry. The development and extent of nephritis were assessed by monitoring proteinuria (using a urine dipstick) and by immunohistochemical analysis of the kidneys. Serum immunoglobulin levels were measured by enzyme‐linked immunosorbent assay.

Results

After a single injection of anti‐mCD20, BCD was more efficient in the peripheral blood, lymph nodes, and spleen compared with the bone marrow and peritoneum of normal mice as well as younger mice with lupus. Since depletion of the marginal zone and peritoneal B cells was incomplete and variable, particularly in older mice with established nephritis, a strategy of sequential weekly dosing was subsequently used, which improved the extent of depletion. BAFF blockade further enhanced depletion in the spleen and lymph nodes. Early BCD therapy delayed disease onset, whereas BCD therapy in mice with advanced disease reduced the progression of nephritis. These effects were long‐lasting, even after B cell reconstitution occurred, and were associated with a reduction in T cell activation but no significant change in autoantibody production.

Conclusion

The lasting benefit of a short course of BCD therapy in lupus‐prone mice with an intact immune system and established disease highlights the validity of this treatment approach.
SIGIRR/TIR‐8 is an inhibitor of toll‐like receptor signaling in primary human cells and regulates inflammation in models of rheumatoid arthritis
Authors: Drexler Stefan K. , Kong Philip , Inglis Julia , Williams Richard O. , Garlanda Cecilia , Mantovani Alberto , Yazdi Amir S. , Brennan Fionula , Feldmann Marc , Foxwell Brian M. J.
Published: April 2010

Objective

Single‐immunoglobulin interleukin‐1 receptor–related (SIGIRR), which is also known as Toll/interleukin‐1 receptor 8 (TIR‐8), is a member of the TIR domain–containing family of receptors and was first characterized as an inhibitor of interleukin‐1 receptor (IL‐1R) and Toll‐like receptor (TLR) signaling. In the Dextran sulfate sodium–induced colitis model, SIGIRR−/− mice were shown to have increased inflammation and to be more susceptible to endotoxin challenge. Increasing evidence implicates TLR and IL‐1R signaling in the pathology of rheumatoid arthritis (RA). Therefore, the purpose of this study was to investigate the involvement of SIGIRR in regulating inflammation in disease‐relevant models.

Methods

Primary human monocyte‐derived macrophages and dendritic cells (DCs) were used to overexpress SIGIRR as well as to knock down endogenously expressed SIGIRR using small interfering RNAs. SIGIRR was also overexpressed in synovial cells derived from RA patients. To investigate the role of SIGIRR in vivo, zymosan‐induced arthritis (ZIA) and collagen antibody–induced arthritis (CAIA) were induced in SIGIRR‐knockout mice.

Results

SIGIRR overexpression inhibited TLR‐induced cytokine production in macrophages and DCs, while SIGIRR knockdown resulted in increased cytokine production following TLR stimulation. Moreover, SIGIRR overexpression inhibited the spontaneous release of cytokines by human RA synovial cells. The role of SIGIRR as an inhibitor of inflammation was confirmed in vivo, since SIGIRR−/− mice developed a more severe disease in both the ZIA and CAIA models.

Conclusion

Our study is the first to show the expression pattern and function of SIGIRR in primary human cells. Furthermore, this investigation defines the role of SIGIRR in disease‐relevant cell types and demonstrates that SIGIRR is a potential therapeutic target for RA.
Expression and function of CXCL16 in a novel model of gout
Authors: Ruth Jeffrey H. , Arendt Monica D. , Amin M. Asif , Ahmed Salahuddin , Marotte Hubert , Rabquer Bradley J. , Lesch Charles , Lee Solhee , Koch Alisa E.
Published: April 2010

Objective

To better define the activity of soluble CXCL16 in the recruitment of polymorphonuclear neutrophils (PMNs) in vivo, utilizing a novel animal model of gout involving engraftment of SCID mice with normal human synovial tissue (ST) injected intragraft with gouty human synovial fluid (SF).

Methods

For in vitro studies, a modified Boyden chemotaxis system was used to identify CXCL16 as an active recruitment factor for PMNs in gouty SF. Migration of PMNs could be reduced by neutralization of CXCL16 activity in gouty SF. For in vivo analyses, fluorescent dye–tagged PMNs were injected intravenously into SCID mice while, simultaneously, diluted gouty SF containing CXCL16, or depleted of CXCL16 by antibody blocking, was administered intragraft. In addition, the receptor for CXCL16, CXCR6, was inhibited by incubating PMNs with a neutralizing anti‐CXCR6 antibody prior to injection into the mouse chimeras. Recruitment of PMNs to the gouty SF–injected normal human ST was then examined in this SCID mouse chimera system.

Results

CXCL16 concentrations were highly elevated in gouty SF, and PMNs were observed to migrate in response to CXCL16 in vitro. Normal human ST–SCID mouse chimeras injected intragraft with gouty SF that had been depleted of CXCL16 during PMN transfer showed a significant reduction of 50% in PMN recruitment to engrafted tissue as compared with that after administration of sham‐depleted gouty SF. Similar findings were achieved when PMNs were incubated with a neutralizing anti‐CXCR6 antibody before injection into chimeras.

Conclusion

Overall, the results of this study outline the effectiveness of the human–SCID mouse chimera system as a viable animal model of gout, serving to identify the primary function of CXCL16 as a significant mediator of in vivo recruitment of PMNs to gouty SF.
Modulation of the angiogenic phenotype of normal and systemic sclerosis endothelial cells by gain–loss of function of pentraxin 3 and matrix metalloproteinase 12
Authors: Margheri Francesca , Serratì Simona , Lapucci Andrea , Chillà Anastasia , Bazzichi Laura , Bombardieri Stefano , Kahaleh Bashar , Calorini Lido , Bianchini Francesca , Fibbi Gabriella , Del Rosso Mario
Published: April 2010

Objective

Studies have shown that in systemic sclerosis (SSc) endothelial cells, overproduction of matrix metalloproteinase 12 (MMP‐12) and pentraxin 3 (PTX3) is associated with defective angiogenesis. This study was undertaken to examine whether overexpression of the relevant molecules could inhibit angiogenesis of normal microvascular endothelial cells (MVECs), and whether silencing of these molecules in SSc MVECs could restore the lost angiogenic properties of the cells in vitro and in vivo.

Methods

Transient transfection of MVECs with human MMP12 and PTX3 was performed by electroporation. Silencing of MMP12 and PTX3 was obtained by treatment with small interfering RNA, and treatment effects were validated by Western blotting with specific antibodies and a fluorimetric assay. In vitro cell migration and capillary morphogenesis were studied on Matrigel substrates. In vivo angiogenesis was studied using a Matrigel sponge assay in mice.

Results

Transfection of MMP12 and PTX3 in normal MVECs resulted in loss of proliferation, invasion, and capillary morphogenesis in vitro, attributed to truncation of the urokinase‐type plasminogen activator receptor by MMP12 and to the anti–fibroblast growth factor 2/anti–vascular endothelial growth factor activity of PTX3. These effects were particularly evident in mixed populations of transfected normal MVECs (50% transfected with MMP12 and 50% with PTX3). Silencing of the same molecules in SSc MVECs increased their invasion in Matrigel. Single‐gene silencing did not increase the capillary morphogenesis of SSc MVECs, whereas double‐gene–silenced cells showed a burst of capillary tube formation. Culture medium of silenced SSc MVECs stimulated angiogenesis in assays of Matrigel sponge invasion in mice.

Conclusion

Overexpression of either MMP12 or PTX3 in normal MVECs blunts their angiogenic properties. Loss of function of MMP12 and PTX3 in SSc MVECs restores the ability of the cells to produce capillaries in vitro and induces vascularization in vivo on a Matrigel sponge.
Intervertebral disc degeneration: New insights based on “skipped” level disc pathology
Authors: Cheung Kenneth M. C. , Samartzis Dino , Karppinen Jaro , Mok Florence P. S. , Ho Daniel W. H. , Fong Daniel Y. T. , Luk Keith D. K.
Published: April 2010

Objective

Typically, age and abnormal physical loading (“wear and tear”) have been associated with the development of intervertebral disc degeneration. In the past decade, various additional etiologic factors for disc degeneration have been sporadically reported in the literature; however, many investigators continue to place tremendous emphasis on the effects of age and biomechanics associated with disc degeneration. The aim of this study was to provide additional insight into the notion that age and biomechanics are key factors in the development of disc degeneration. To this end, we addressed the prevalence of and risk factors associated with a unique pattern of disc degeneration of the lumbar spine, “skipped” level (nonconsecutive) disc degeneration (SLDD).

Methods

As part of a large genetics‐based study in southern Chinese individuals (n = 1,989), a cross‐sectional analysis was performed in subjects exhibiting disc degeneration in ≥2 levels (n = 838) who were then categorized as having SLDD (n = 174) or non‐SLDD (contiguous, multilevel; n = 664). Various radiographic parameters were assessed based on T2‐weighted magnetic resonance imaging (MRI). Subject demographics were assessed, and univariate and multivariate logistic regression analyses were performed.

Results

Overall, 8.7% of the whole population (n = 1,989) had SLDD, while it was present in 20.8% of subjects with multilevel disc degeneration (n = 838). SLDD was more prevalent in male subjects (adjusted odds ratio [OR] 1.48, 95% confidence interval [95% CI] 1.04–2.10, P = 0.028). SLDD was significantly associated with the presence of Schmorl's nodes (adjusted OR 2.72, 95% CI 1.78–4.15, P < 0.001), which also presented in levels with no disc degeneration. A history of disc bulge/extrusion (P = 0.004) and/or a history of back injury (P = 0.010) was significantly associated with non‐SLDD, and a greater degree of overall severity of disc degeneration was also associated with non‐SLDD. Other demographic and MRI findings did not significantly differ between groups.

Conclusion

To our knowledge, this report is the first to describe the prevalence and risk factors associated with SLDD. Our study challenges the paradigm that age and biomechanics are the key factors associated with the development of disc degeneration. Although age and biomechanical factors may play a role in the manifestation of disc degeneration, our novel findings of SLDD patterns provide further awareness of and support for the notion that additional etiologic factors may play a role in the development of disc degeneration. Such factors warrant further investigation to shed light on the cause of disc degeneration.
Ofatumumab, a human anti‐CD20 monoclonal antibody, for treatment of rheumatoid arthritis with an inadequate response to one or more disease‐modifying antirheumatic drugs: Results of a randomized, double‐blind, placebo‐controlled, phase I/II study
Authors: Østergaard Mikkel , Baslund Bo , Rigby William , Rojkovich Bernadette , Jorgensen Christian , Dawes Peter T. , Wiell Charlotte , Wallace Daniel J. , Tamer Søren C. , Kastberg Helle , Petersen Jørgen , Sierakowski Stanislaw
Published: April 2010

Objective

To investigate the safety and efficacy of ofatumumab, a novel human anti‐CD20 monoclonal antibody (mAb), in patients with active rheumatoid arthritis (RA) whose disease did not respond to ≥1 disease‐modifying antirheumatic drug.

Methods

This combined phase I/II study investigated the safety and efficacy of 3 doses of ofatumumab. In part A (phase I), 39 patients received 2 intravenous (IV) infusions of ofatumumab (300 mg, 700 mg, or 1,000 mg) or placebo in a 4:1 ratio 2 weeks apart, using a specified premedication and infusion regimen. In part B (phase II), 225 patients received study treatment as per phase I in a 1:1:1:1 ratio. Safety was assessed by adverse events (AEs) and laboratory parameters. Efficacy was assessed by the American College of Rheumatology 20% criteria for improvement (ACR20), the Disease Activity Score in 28 joints, and the European League Against Rheumatism (EULAR) response criteria. B cell pharmacodynamics were also investigated.

Results

AEs were predominantly reported at the first infusion and were mostly mild to moderate in intensity. Rapid and sustained peripheral B cell depletion was observed in all dose groups. In phase II, patients in all ofatumumab dose groups had significantly higher ACR20 response rates (40%, 49%, and 44% for the 300 mg, 700 mg, and 1,000 mg doses, respectively) than did patients receiving placebo (11%) at week 24 (P < 0.001). Overall, 70% of patients receiving ofatumumab had a moderate or good response according to the EULAR criteria at week 24.

Conclusion

Our findings indicate that ofatumumab, administered as 2 IV infusions of doses up to 1,000 mg, is clinically effective in patients with active RA.
Mapping the geography of science: Distribution patterns and networks of relations among cities and institutes
Authors: Leydesdorff Loet , Persson Olle
Published: April 2010

Using Google Earth, Google Maps, and/or network visualization programs such as Pajek, one can overlay the network of relations among addresses in scientific publications onto the geographic map. The authors discuss the pros and cons of various options, and provide software (freeware) for bridging existing gaps between the Science Citation Indices (Thomson Reuters) and Scopus (Elsevier), on the one hand, and these various visualization tools on the other. At the level of city names, the global map can be drawn reliably on the basis of the available address information. At the level of the names of organizations and institutes, there are problems of unification both in the ISI databases and with Scopus. Pajek enables a combination of visualization and statistical analysis, whereas the Google Maps and its derivatives provide superior tools on the Internet.
Robust l2−l∞ state feedback control for uncertain discrete‐time switched systems with mode‐dependent time‐varying delays
Authors: Zong Guangdeng , Hou Linlin , Xu Shengyuan
Published: April 2010

This paper deals with the problem of robust l2−l∞ state feedback control for uncertain discrete‐time switched systems with mode‐dependent time‐varying delays. Attention is focused on the design of a switched state feedback controller, which guarantees the asymptotical stability of the closed‐loop system and reduces the effect of the disturbance input on the controlled output to a prescribed level for all admissible uncertainties. By resorting to a descriptor system approach, delay‐dependent sufficient conditions are presented in terms of linear matrix inequalities (LMIs). A numerical example is provided to demonstrate the effectiveness of the proposed algorithms.

Copyright © 2010 John Wiley and Sons Asia Pte Ltd and Chinese Automatic Control Society
Diagnosis and treatment of small follicular thyroid carcinomas
Authors: Clerici T , Kolb W , Beutner U , Bareck E , Dotzenrath C , Kull C , Niederle B
Published: April 2010

Background

Follicular thyroid microcarcinomas (mFTCs) of 10 mm or less in size rarely manifest clinically and their clinical significance is controversial. This study assessed their characteristics and incidence, and analysed treatment modalities used for mFTC.

Methods

Members of the German Association of Endocrine Surgeons were asked to review patients with mFTC operated on between 1990 and 2005.

Results

Data for 90 patients from 26 institutions were reported. Histopathological slides were available for re-evaluation in 35 patients. Most initial diagnoses had to be revised because of incorrect size assessment or incorrect diagnosis (benign adenoma, papillary thyroid carcinoma (PTC), follicular variant of PTC). The diagnosis of mFTC was confirmed in only four patients. As a result of the incorrect histopathological diagnosis, unnecessary completion thyroidectomy and radioiodine ablation were performed in 17 and 20 patients respectively. The incidence of mFTC was calculated to be 0·12 per million population per year.

Conclusion

mFTC is exceptionally rare. Such tumours are overdiagnosed, resulting in unnecessary treatment associated with avoidable morbidity. Histopathological re-evaluation by an experienced pathologist is recommended before embarking on further treatments when a diagnosis of mFTC is made.
Patterns of recurrence in patients with acute diverticulitis
Authors: Eglinton T , Nguyen T , Raniga S , Dixon L , Dobbs B , Frizelle F A
Published: April 2010

Background

The natural history of acute diverticulitis remains unclear, with the role of prophylactic surgery following conservatively managed diverticulitis increasingly controversial. This study investigated recurrence rates, patterns and complications after conservatively managed diverticulitis.

Methods

This was a retrospective chart review of all patients admitted with diverticulitis between June 1997 and June 2002. Demographic data, management, recurrence rates, complications and subsequent surgery were recorded.

Results

Some 502 patients were identified, 337 with uncomplicated and 165 with complicated diverticulitis. Median follow-up was 101 (range 60–124) months. Of 320 patients with uncomplicated diverticulitis managed conservatively, 60 (18·8 per cent) had one episode of recurrence, whereas 15 (4·7 per cent) had two or more episodes. After an initial attack of uncomplicated diverticulitis, only 5·0 per cent developed complicated disease. Complicated disease recurred in 24 per cent, compared with a recurrence rate of 23·4 per cent in those with uncomplicated diverticulitis (P = 0·622). When recurrence occurred, it usually did so within 12 months of the initial episode.

Conclusion

Acute diverticulitis has a low recurrence rate and rarely progresses to complications. Any recurrence is usually early, in a pattern more consistent with failure of the index episode to settle. Subsequent elective surgery to prevent recurrence and the development of complications should be used sparingly.
A national propensity‐adjusted analysis of adjuvant radiotherapy in the treatment of resected pancreatic adenocarcinoma
Authors: McDade Theodore P. , Hill Joshua S. , Simons Jessica P. , Piperdi Bilal , Ng Sing Chau , Zhou Zheng , Kadish Sidney P. , Fitzgerald Thomas J. , Tseng Jennifer F.
Published: April 2010

BACKGROUND:

The benefit of adjuvant radiotherapy (RT) for resected pancreatic adenocarcinoma remains controversial after randomized clinical trials. In this national‐level US study, a propensity score (conditional probability of receiving RT) was used to adjust for potential confounding in nonrandomized designs from treatment group differences.

METHODS:

Patients were identified from the Surveillance, Epidemiology, and End Results (SEER) registry (1988‐2005 dataset). Multivariate analyses to determine the effect of RT on overall survival were performed using propensity‐adjusted Cox proportional hazards and Kaplan‐Meier analyses.

RESULTS:

In total, 5676 patients with resected pancreatic adenocarcinoma were identified, and 40.8% of those patients had received adjuvant RT. Univariate predictors of survival included age, race, marital status, disease stage, tumor size, tumor extension, tumor grade, lymph node status, year of diagnosis, type of resection, and receipt of RT (all P < .002). In a Cox model, independent predictors of improved survival included white race, married status, earlier stage, smaller tumors, well differentiated tumors, negative lymph node (N0) status, recent diagnosis, and receipt of RT (all P < .05). In a propensity‐adjusted proportional hazards regression, the benefit of adjuvant treatment that included RT remained significant after adjusting for the likelihood of receiving RT (hazard ratio, 0.773; 95% confidence interval, 0.714‐0.836; P < .0001). Within all 5 propensity strata, Kaplan‐Meier survival differed significantly (P < .0001 [lowest and highest probability strata] and P = .0165 [middle stratum with a “pseudorandom” probability of RT]).

CONCLUSIONS:

Adjuvant RT for resected pancreatic adenocarcinoma was associated with a significant survival advantage in a large national database, even after using propensity score methods to adjust for differences between treatment groups. The authors concluded that adjuvant RT should be considered for all appropriate patients who have resected pancreatic adenocarcinoma. Cancer 2010. © 2010 American Cancer Society.
Signaling of ERBB receptor tyrosine kinases promotes neuroblastoma growth in vitro and in vivo
Authors: Richards Kristen N. , Zweidler‐McKay Patrick A. , Van Roy Nadine , Speleman Frank , Trevino Jesus , Zage Peter E. , Hughes Dennis P. M.
Published: April 2010

BACKGROUND:

ERBB receptor tyrosine kinases can mediate proliferation, migration, adhesion, differentiation, and survival in many types of cells and play critical roles in many malignancies. Recent reports suggest a role for EGFR signaling in proliferation and survival of neuroblastoma, a common form of pediatric cancer that often has an extremely poor outcome.

METHODS:

The authors examined ERBB family expression in neuroblastoma cell lines and patient samples by flow cytometry, western blot, and quantitative real time polymerase chain reaction (Q‐PCR). Response to ERBB inhibition was assessed in vitro by cell‐cycle analysis and western blot and in vivo by serial tumor‐size measurements.

RESULTS:

A panel of neuroblastoma cell lines and primary patient tumors expressed EGFR, HER‐3, and HER‐4, with HER‐2 in some tumors.HER‐4mRNA was expressed predominantly in cleavable isoforms. Whereas EGFR inhibition with erlotinib and pan‐ERBB inhibition with CI‐1033 inhibited EGF‐induced phosphorylation of EGFR, AKT, and ERK1/2, only CI‐1033 induced growth inhibition and dose‐dependent apoptosis in vitro. Both CI‐1033 and erlotinib treatment of neuroblastoma xenograft tumors resulted in decreased tumor growth in vivo, although CI‐1033 was more effective. In vivo expression of EGFR was observed predominantly in vascular endothelial cells.

CONCLUSIONS:

Pan‐ERBB inhibition is required for ERBB‐related neuroblastoma apoptosis in vitro, although EGFR contributes indirectly to tumor growth in vivo. Inhibition of EGFR in endothelial cells may be an important aspect of erlotinib's impact on neuroblastoma growth in vivo. Our results suggest that non‐EGFR ERBB family members contribute directly to neuroblastoma growth and survival, and pan‐ERBB inhibition represents a potential therapeutic target for treating neuroblastoma. Cancer 2010. © 2010 American Cancer Society.
Cloaking via change of variables for the Helmholtz equation
Authors: Kohn Robert V. , Onofrei Daniel , Vogelius Michael S. , Weinstein Michael I.
Published: April 2010

The transformation optics approach to cloaking uses a singular change of coordinates, which blows up a point to the region being cloaked. This paper examines a natural regularization, obtained by (1) blowing up a ball of radius ρ rather than a point, and (2) including a well‐chosen lossy layer at the inner edge of the cloak. We assess the performance of the resulting near‐cloak as the regularization parameter ρ tends to 0, in the context of (Dirichlet and Neumann) boundary measurements for the time‐harmonic Helmholtz equation. Since the goal is to achieve cloaking regardless of the content of the cloaked region, we focus on estimates that are uniform with respect to the physical properties of this region. In three space dimensions our regularized construction performs relatively well: the deviation from perfect cloaking is of order ρ. In two space dimensions it does much worse: the deviation is of order 1/|log ρ|. In addition to proving these estimates, we give numerical examples demonstrating their sharpness. Some authors have argued that perfect cloaking can be achieved without losses by using the singular change‐of‐variable‐based construction. In our regularized setting the analogous statement is false: without the lossy layer, there are certain resonant inclusions (depending in general on ρ) that have a huge effect on the boundary measurements. © 2010 Wiley Periodicals, Inc.
CD4+CD25+ regulatory T cells utilize FasL as a mechanism to restrict DC priming functions in cutaneous immune responses
Authors: Gorbachev Anton V. , Fairchild Robert L.
Published: April 2010

Recent studies have suggested Fas‐mediated elimination of antigen‐presenting cells as an important mechanism down‐regulating the induction of autoimmune responses. It remains unknown whether this mechanism restricts the magnitude of immune responses to non‐self antigens. We used a mouse model of a cutaneous CD8+ T‐cell‐mediated immune response (contact hypersensitivity, CHS) to test if CD4+CD25+ T cells expressing FasL regulate hapten‐specific effector CD8+ T cell expansion through the elimination of Fas‐expressing hapten‐presenting DC. In WT mice, attenuation of CD4+CD25+ T regulatory cell activity by anti‐CD25 mAb increased hapten‐presenting DC numbers in skin‐draining LN, which led to increased effector CD8+ T‐cell priming for CHS responses. In contrast, CD4+CD25+ T cells did not regulate hapten‐specific CD8+ T‐cell priming and CHS responses initiated by Fas‐defective (lpr) DC. Thus, restricting DC priming functions through Fas–FasL interactions is a potent mechanism employed by CD4+CD25+ regulatory cells to restrict CD8+ T‐cell‐mediated allergic immune responses in the skin.
Steady‐state antigen‐expressing dendritic cells terminate CD4+ memory T‐cell responses
Authors: Nasreen Mariam , Waldie Tanya M. , Dixon Chantelle M. , Steptoe Raymond J.
Published: April 2010

CD4+ T cells are important effectors of inflammation and tissue destruction in many diseases of immune dysregulation. As memory T cells develop early during the preclinical stages of autoimmune and inflammatory diseases, immunotherapeutic approaches to treatment of these diseases, once established, must include the means to terminate memory T‐cell responses. Traditionally, it has been considered that, due to their terminally differentiated nature, memory T cells are resistant to tolerance induction, although emerging evidence indicates that some immunotherapeutic approaches can terminate memory T‐cell responses. Here, we demonstrate that CD4+ memory T‐cell responses can be terminated when cognate antigen is transgenically expressed in steady‐state DC. Transfer of in‐vitro‐generated CD4+ memory T cells establishes, in nontransgenic recipients, a stable and readily recalled memory response to cognate antigen. In contrast, upon transfer to mice expressing cognate antigen targeted to DC, memory CD4+ T cells undergo a phase of limited proliferation followed by substantial deletion, and recall responses are effectively silenced. This finding is important in understanding how to effectively apply immunotherapy to ongoing T‐cell‐mediated autoimmune and inflammatory diseases.
Helminths and dendritic cells: Sensing and regulating via pattern recognition receptors, Th2 and Treg responses
Authors: Everts Bart , Smits Hermelijn H. , Hokke Cornelis H. , Yazdanbakhsh Maria
Published: April 2010

The classical reaction of the host to helminth infections is the induction of Th2 immune responses with a regulatory component. DC, as central players in the induction and maintenance of immune responses, play a prominent role in both these processes, and in recent years considerable progress has been made in elucidating the mechanisms behind the interplay between DC and helminths. It is becoming increasingly clear that helminths modulate DC function not only via direct interactions but also indirectly via host‐derived cues. Furthermore, while studies have until recently focused on receptor signaling‐mediated DC modulation by helminths, evidence is emerging that DC may also respond to helminth infections by sensing stress signals or tissue damage inflicted by the worms or their products. Here, we will discuss these new insights and will link them to the origin and importance of Th2 and regulatory immune responses with respect to the survival of both parasite and host.
Production and applications of trehalose lipid biosurfactants
Authors: Franzetti Andrea , Gandolfi Isabella , Bestetti Giuseppina , Smyth Thomas J. P. , Banat Ibrahim M.
Published: April 2010

Different types of trehalose containing glycolipids are known to be produced by several microorganisms belonging to the mycolates group, such as Mycobacterium, Rhodococcus, Arthrobacter, Nocardia and Gordonia. Different structures have been elucidated particularly in Rhodococcus genus. Trehalolipids have gained increased interest for their potential applications in a number of fields due to their ability to lower interfacial tension and increase pseudosolubility of hydrophobic compounds. The most widespread application is in bioremediation technologies as such compounds are known to enhance bioavailability of hydrocarbons. In comparison to other microbial glycolipids, trehalolipids have generally showed contrasting results and achievements with both cases of inhibition and enhancement of biodegradation rates. One of the important challenges regarding potential use of trehalose lipids in a variety of applications is the optimisation of their production and downstream processing. In fact, the purification of the target biological compounds by downstream processing can account for over half the production cost in many biotechnology applications. This is especially true in the case of the Rhodococcal glycolipids, which are often bound to cellular envelopes and are usually produced along with other surface active lipids. In this review, we highlight the current knowledge of trehalolipid biosurfactant applications and the latest successful strategies employed to reduce the cost of their production.
Peanut oil: Compositional data
Authors: Carrín María E. , Carelli Amalia A.
Published: April 2010

The major fatty acids of peanut oil acylglycerols are palmitic (C16:0), oleic (C18:1), and linoleic (C18:2) acids, and only a trace amount of linolenic fatty acid (C18:3) is present. Thus they have a very convenient oxidative stability and have been considered premium cooking and frying oils. This paper provides information about compositional data of peanut oil taking into account major (triacylglycerols and their fatty acids) and minor (free fatty acids, diacylglycerols, phospholipids, sterols, tocopherols, tocotrienols, triterpenic and aliphatic alcohols, waxes, pigments, phenolic compounds, volatiles, and metals) compounds. Moreover, the influence of genotype, seed maturity, climatic conditions, and growth location on peanut oil chemical composition is considered in the present report. In addition, peanut oils from wild species found in South America as well as from peanut lines developed through conventional breeding are also compared.
Supplementation of tocotrienol‐rich fraction increases interferon‐gamma production in ovalbumin‐immunized mice
Authors: Nesaretnam Kalanithi , Mahalingam Dashayini , Radhakrishnan Ammu Kutty , Premier Robert
Published: April 2010

Palm oil is a rich source of vitamin E. The tocotrienol‐rich fraction (TRF) extracted from palm oil contain 70% tocotrienols and 30% tocopherols. The effect of TRF supplementation on the immune modulation was evaluated in 6‐wk‐old female BALB/c mice immunized with ovalbumin (OVA) adjuvanted with alum. Mice in control and experimental groups were immunized subcutaneously (s.c.) on days 14 and 28 with a single dose of 50 µg OVA. The mice in the experimental group were orally gavaged daily with 1 mg of TRF from palm oil while those in the control group received carrier oil. The results show that mice in the experimental group produced significantly (p<0.05) higher levels of interferon‐gamma (IFN‐γ) compared to the control group. There was no significant (p>0.05) difference in the levels of interleukin‐4 (IL‐4) produced between the control and experimental animals. Lymphocyte proliferation in response to mitogen or OVA stimulation was significantly (p<0.05) higher in splenocytes derived from the TRF supplemented mice compared to control mice. These findings show that daily supplementation of palm TRF can induce a strong cell‐mediated immune response, i.e., T‐helper‐1 (Th1) response, which would be beneficial to fight viral infections and cancer.
Yarrowia lipolytica: A model microorganism used for the production of tailor‐made lipids
Authors: Papanikolaou Seraphim , Aggelis George
Published: April 2010

This paper presents a review of studies related with the production of specialty lipids by the yeastYarrowia lipolyticaduring growth on various fatty agro‐industrial residues utilized as substrates. In a first approach, the potential for growth on stearin (a low‐cost industrial derivative of tallow composed of saturated free fatty acids) was assessed; significant biomass production was obtained, accompanied by notable intracellular accumulation of lipid which occurred as a primary anabolic activity regardless of the extracellular nitrogen availability in the medium.Y. lipolyticaconsumed its own storage lipids in the stationary growth phase. The secretion of extracellular lipase in non‐negligible quantities was also observed in cultures performed on stearin. AlthoughY. lipolyticatended to increase the intracellular level of stearic acid, its reserve lipid could be used as a cocoa butter substitute, provided that an oleic acid donor, such as chemically hydrolyzed rapeseed oil, is used as co‐substrate together with stearin. Accordingly, predetermined lipids with composition similarities to cocoa butter were synthesized using low‐cost substrates. Other strategies related with the production of microbial cocoa butter substitutes are also presented and comprehensively discussed.
Association of Apo E gene polymorphism with HDL level in Tehranian population
Authors: Daneshpour Maryam S. , Hedayati Mehdi , Eshraghi Parisa , Azizi Fereidoun
Published: April 2010

The prevalence of cardiovascular risk factors, especially low level of high density lipoprotein cholesterol (HDL‐C), is very high in Iran. The associations of apolipoprotein E (Apo E) polymorphism with lipid profile, especially high density lipoprotein (HDL) subfractions, were examined in Iranian population. A cross‐sectional study of 1030 subjects (452 men and 578 women) from the Tehran Lipid and Glucose Study (TLGS) was performed. Serum triacylglycerol, cholesterol, FBS, HDL‐C levels and its subfractions, Apo B and Apo A1 were determined and body mass index and blood pressure were measured. A segment of the mentioned gene was amplified by PCR and the polymorphisms were revealed by RFLP using HhaI restriction enzyme. Allele frequencies obtained for APOE*2, APOE*3, and APOE*4 were 5.77, 85.92, and 8.3%, respectively. The presence of the ε2 allele was significantly associated with increased serum HDL‐C levels and its subfractions both in men and women except HDL3 in men. The LDL/HDL ratio was significantly lower in female. The relations were significant even after adjustment for age, sex, and BMI but hypertension, smoking, and diabetes status decreased the effect of Apo E2 on HDL‐C and HDL subfractions. The observed genotype and allele frequencies were similar to those reported for other Caucasians samples. Apo E2 increased the level of HDL‐C, HDL subfractions, and decreased the LDL/HDL ratio. These findings highlight the important effect of variation in this gene on lipid levels.
Composition and physical properties of arugula, shepherd's purse, and upland cress oils
Authors: Moser Bryan R. , Winkler‐Moser Jill K. , Shah Shailesh N. , Vaughn Steven F.
Published: April 2010

The fatty acid, tocopherol, and phytosterol profiles of arugula (AO; Eruca vesicaria (L.) Cav. subsp. sativa (Mill.) Thell.), upland cress (UCO; Barbarea verna (Mill.) Asch.), and shepherd's purse (SPO; Capsella bursa‐pastoris (L.) Medik.) oils are reported, along with their physical properties. The tocopherol content of SPO (770 mg/kg) was higher than detected in AO (656 mg/kg) and UCO (430 mg/kg). UCO contained a higher concentration of phytosterols (8.47 mg/g) than SPO (7.19 mg/g) and AO (6.60 mg/g). The oil contents of AO, SPO, and UCO were 27.0, 26.6, and 24.2 wt%. The acid values of AO (0.54 mg KOH/g) and UCO (0.32 mg KOH/g) were lower than obtained for SPO (3.19 mg KOH/g). The principle fatty acid in AO and UCO was erucic acid (41.7 and 36.8 wt%), whereas SPO contained primarily linolenic (32.4 wt%) and linoleic (20.5 wt%) acids. AO and UCO were more stable to oxidation than SPO, as indicated by induction periods (110°C; EN 14 112) of 16.5, 7.9, and 2.2 h. SPO exhibited lower kinematic viscosity and higher viscosity index values than AO and UCO. UCO displayed the lowest pour point (−29°C), followed by AO (−22°C) and SPO (−14°C).