bluuebunny/crossref_metadata_2025 huggingface dataset:
Mammalian Target of Rapamycin Inhibition and Alloantigen-Specific Regulatory T Cells Synergize To Promote Long-Term Graft Survival in Immunocompetent Recipients
Authors: Raimondi Giorgio , Sumpter Tina L. , Matta Benjamin M. , Pillai Mahesh , Corbitt Natasha , Vodovotz Yoram , Wang Zhiliang , Thomson Angus W.
Published: December 2009

Minimization of immunosuppression and donor-specific tolerance to MHC-mismatched organ grafts are important clinical goals. The therapeutic potential of regulatory T cells (Tregs) has been demonstrated, but conditions for optimizing their in vivo function posttransplant in nonlymphocyte-depleted hosts remain undefined. In this study, we address mechanisms through which inhibition of the mammalian target of rapamycin (Rapa) synergizes with alloantigen-specific Treg (AAsTreg) to permit long-term, donor-specific heart graft survival in immunocompetent hosts. Crucially, immature allogeneic dendritic cells allowed AAsTreg selection in vitro, with minimal expansion of unwanted (Th17) cells. The rendered Treg potently inhibited T cell proliferation in an Ag-specific manner. However, these AAsTreg remained unable to control T cells stimulated by allogeneic mature dendritic cells, a phenomenon dependent on the release of proinflammatory cytokines. In vivo, Rapa administration reduced danger-associated IL-6 production, T cell proliferation, and graft infiltration. Based on these observations, AAsTreg were administered posttransplant (day 7) in combination with a short course of Rapa and rendered >80% long-term (>150 d) graft survival, a result superior to that achieved with polyclonal Treg. Moreover, graft protection was alloantigen-specific. Significantly, long-term graft survival was associated with alloreactive T cell anergy. These findings delineate combination of transient mammalian target of Rapa inhibition with appropriate AAsTreg selection as an effective approach to promote long-term organ graft survival.
Dual Control of Antitumor CD8 T Cells through the Programmed Death-1/Programmed Death-Ligand 1 Pathway and Immunosuppressive CD4 T Cells: Regulation and Counterregulation
Authors: Currie Andrew J. , Prosser Amy , McDonnell Alison , Cleaver Amanda L. , Robinson Bruce W. S. , Freeman Gordon J. , van der Most Robbert G.
Published: December 2009

Tumors have evolved multiple mechanisms to evade immune destruction. One of these is expression of T cell inhibitory ligands such as programmed death-ligand 1 (PD-L1; B7-H1). In this study, we show that PD-L1 is highly expressed on mesothelioma tumor cells and within the tumor stroma. However, PD-L1 blockade only marginally affected tumor growth and was associated with the emergence of activated programmed death-1+ ICOS+ CD4 T cells in tumor-draining lymph nodes, whereas few activated CD8 T cells were present. Full activation of antitumor CD8 T cells, characterized as programmed death-1+ ICOS+ Ki-67+ and displaying CTL activity, was only observed when CD4 T cells were depleted, suggesting that a population of suppressive CD4 T cells exists. ICOS+ foxp3+ regulatory T cells were found to be regulated through PD-L1, identifying one potentially suppressive CD4 T cell population. Thus, PD-L1 blockade activates antitumor CD8 T cell most potently in the absence of CD4 T cells. These findings have implications for the development of PD-L1-based therapies.
Inhibition of IL-17A Attenuates Atherosclerotic Lesion Development in ApoE-Deficient Mice
Authors: Erbel Christian , Chen Lili , Bea Florian , Wangler Susanne , Celik Sultan , Lasitschka Felix , Wang Ying , Böckler Dittmar , Katus Hugo A. , Dengler Thomas J.
Published: December 2009

The importance of an (auto)immune response in atherogenesis is becoming increasingly well understood. IL-17A-expressing T cells modulate immune cell trafficking, initiating inflammation and cytokine production in (auto)immune diseases. In human carotid artery plaques, we previously showed the presence of IL-17A-producing T cells and IL-23; however, IL-17A effects on atherogenesis have not been studied. Aortic root sections from 8-wk-old apolipoprotein E-deficient mice fed a standard chow diet were examined after 12 wk for lesion area, plaque composition, cellular infiltration, cytokine expression, and apoptosis. The treatment group (n = 15) received anti-IL-17A Ab and the controls (n = 10) received irrelevant Abs. Inhibition of IL-17A markedly reduced atherosclerotic lesion area (p < 0.001), maximal stenosis (p < 0.001), and vulnerability of the lesion. IL-17A mAb-treated mice showed reduced cellular infiltration, down-regulation of activation markers on endothelium and immune cells (e.g., VCAM-1), and reduced cytokine/chemokine secretion (e.g., IL6, TNFα, CCL5). To investigate possible mechanisms, different atherogenic cell types (e.g., macrophages, dendritic cells, HUVECs, vascular smooth muscle cells) were stimulated with IL-17A in addition to TNF-α, IFN-γ, or LPS to induce cellular activation or apoptosis in vitro. Stimulation with IL-17A induced proinflammatory changes in several atherogenic cell types and apoptotic cell death in murine cells. Functional blockade of IL-17A reduces atherosclerotic lesion development and decreases plaque vulnerability, cellular infiltration, and tissue activation in apolipoprotein E-deficient mice. The present data support a pathogenic role of IL-17A in the development of atherosclerosis by way of its widespread proinflammatory and proapoptotic effects on atherogenic cells.
Crucial Role of Phospholipase Cε in Induction of Local Skin Inflammatory Reactions in the Elicitation Stage of Allergic Contact Hypersensitivity
Authors: Hu Lizhi , Edamatsu Hironori , Takenaka Nobuyuki , Ikuta Shuzo , Kataoka Tohru
Published: December 2009

Phospholipase Cε (PLCε) is an effector of Ras/Rap small GTPases. We previously demonstrated that PLCε plays a crucial role in development of phorbor ester-induced skin inflammation, which is intimately involved in the promotion of skin carcinogenesis. In this study, we have examined its role in local skin inflammatory reactions during development of contact hypersensitivity toward a hapten 2,4-dinitrofluorobenzene (DNFB). PLCε+/+ and PLCε−/− mice were sensitized with DNFB, followed by a DNFB challenge on the ears. PLCε−/− mice exhibited substantially attenuated inflammatory reactions compared with PLCε+/+ mice as shown by suppression of ear swelling, neutrophil infiltration, and proinflammatory cytokine production. In contrast, the extent and kinetics of CD4+ T cell infiltration showed no difference depending on the PLCε background. Adoptive transfer of CD4+ T cells from the sensitized mice to naive mice between PLCε+/+ and PLCε−/− backgrounds indicated that PLCε exerts its function in cells other than CD4+ T cells, presumably fibroblasts or keratinocytes of the skin, to augment inflammatory reactions during the elicitation stage of contact hypersensitivity. Moreover, dermal fibroblasts and epidermal keratinocytes cultured from the skin expressed proinflammatory cytokines in a PLCε-dependent manner on stimulation with T cell-derived cytokines such as IL-17, IFN-γ, TNF-α, and IL-4. These results indicate that PLCε plays a crucial role in induction of proinflammatory cytokine expression in fibroblasts and keratinocytes at the challenged sites, where infiltrated CD4+ T cells produce their intrinsic cytokines, thereby augmenting the local inflammatory reactions.
Cell Surface Externalization of Annexin A1 as a Failsafe Mechanism Preventing Inflammatory Responses during Secondary Necrosis
Authors: Blume Karin E. , Soeroes Szabolcs , Waibel Michaela , Keppeler Hildegard , Wesselborg Sebastian , Herrmann Martin , Schulze-Osthoff Klaus , Lauber Kirsten
Published: December 2009

The engulfment of apoptotic cells is of crucial importance for tissue homeostasis in multicellular organisms. A failure of this process results in secondary necrosis triggering proinflammatory cytokine production and autoimmune disease. In the present study, we investigated the role of annexin A1, an intracellular protein that has been implicated in the efficient removal of apoptotic cells. Consistent with its function as bridging protein in the phagocyte synapse, opsonization of apoptotic cells with purified annexin A1 strongly enhanced their phagocytic uptake. A detailed analysis, however, surprisingly revealed that annexin A1 was hardly exposed to the cell surface of primary apoptotic cells, but was strongly externalized only on secondary necrotic cells. Interestingly, while the exposure of annexin A1 failed to promote the uptake of these late secondary necrotic cells, it efficiently prevented induction of cytokine production in macrophages during engulfment of secondary necrotic cells. Our results therefore suggest that annexin A1 exposure during secondary necrosis provides an important failsafe mechanism counteracting inflammatory responses, even when the timely clearance of apoptotic cells has failed.
Asynchronous RAG-1 Expression during B Lymphopoiesis
Authors: Welner Robert S. , Esplin Brandt L. , Garrett Karla P. , Pelayo Rosana , Luche Hervé , Fehling Hans Jörg , Kincade Paul W.
Published: December 2009

Changes in cell surface markers and patterns of gene expression are commonly used to construct sequences of events in hematopoiesis. However, the order may not be as rigid as once thought and it is unclear which changes represent the best milestones of differentiation. We developed a fate-mapping model where cells with a history of RAG-1 expression are permanently marked by red fluorescence. This approach is valuable for appreciating lymphoid-lineage relationships without need for irradiation and transplantation. Hematopoietic stem cells (HSC) as well as myeloid and dendritic cell progenitors were unlabeled. Also as expected, most previously identified RAG-1+ early lymphoid progenitors in bone marrow and all lymphoid-affiliated cells were marked. Of particular interest, there was heterogeneity among canonical common lymphoid progenitors (CLP) in bone marrow. Labeled CLP expressed slightly higher levels of IL-7Rα, displayed somewhat less c-Kit, and generated CD19+ lymphocytes faster than the unlabeled CLP. Furthermore, CLP with a history of RAG-1 expression were much less likely to generate dendritic and NK cells. The RAG-1-marked CLP were lineage stable even when exposed to LPS, while unlabeled CLP were redirected to become dendritic cells in response to this TLR4 ligand. These findings indicate that essential events in B lymphopoiesis are not tightly synchronized. Some progenitors with increased probability of becoming lymphocytes express RAG-1 while still part of the lineage marker-negative Sca-1+c-Kithigh (LSK) fraction. Other progenitors first activate this locus after c-Kit levels have diminished and cell surface IL-7 receptors are detectable.
The Kinase Inhibitors Sunitinib and Sorafenib Differentially Affect NK Cell Antitumor Reactivity In Vitro
Authors: Krusch Matthias , Salih Julia , Schlicke Manuela , Baessler Tina , Kampa Kerstin Maria , Mayer Frank , Salih Helmut Rainer
Published: December 2009

Sunitinib and Sorafenib are protein kinase inhibitors (PKI) approved for treatment of patients with advanced renal cell cancer (RCC). However, long-term remissions of advanced RCC have only been observed after IL-2 treatment, which underlines the importance of antitumor immune responses in RCC patients. Because PKI, besides affecting tumor cells, also may inhibit signaling in immune effector cells, we determined how Sunitinib and Sorafenib influence antitumor immunity. We found that cytotoxicity and cytokine production of resting and IL-2-activated PBMC are inhibited by pharmacological concentrations of Sorafenib but not Sunitinib. Analysis of granule-mobilization within PBMC revealed that this was due to impaired reactivity of NK cells, which substantially contribute to antitumor immunity by directly killing target cells and shaping adaptive immune responses by secreting cytokines like IFN-γ. Analyses with resting and IL-2-activated NK cells revealed that both PKI concentration dependently inhibit cytotoxicity and IFN-γ production of NK cells in response to tumor targets. This was due to impaired PI3K and ERK phosphorylation which directly controls NK cell reactivity. However, while Sorafenib inhibited NK cell effector functions and signaling at levels achieved upon recommended dosing, pharmacological concentrations of Sunitinib had no effect, and this was observed upon stimulation of NK cell reactivity by tumor target cells and upon IL-2 treatment. In light of the important role of NK cells in antitumor immunity, and because multiple approaches presently aim to combine PKI treatment with immunotherapeutic strategies, our data demonstrate that choice and dosing of the most suitable PKI in cancer treatment requires careful consideration.
Inflammasome-Associated Nucleotide-Binding Domain, Leucine-Rich Repeat Proteins and Inflammatory Diseases
Authors: Jha Sushmita , Ting Jenny P.-Y.
Published: December 2009

The nucleotide-binding domain, leucine-rich repeat (NLR) proteins are a recently discovered family of intracellular pathogen and danger signal sensors. NLRs have emerged as important contributors to innate immunity in animals. The physiological impact of these genes is increasingly evident, underscored by the genetic association of variant family members with an array of inflammatory diseases. The association of mutations in NLR genes with autoinflammatory diseases indicates an important function of these genes in inflammation in vivo. This review summarizes the role of the inflammasome NLR proteins in innate immunity and inflammatory diseases and explores the possible utility of some of these NLRs as pharmacological targets.
Allele *1 of HS1.2 Enhancer Associates with Selective IgA Deficiency and IgM Concentration
Authors: Giambra Vincenzo , Cianci Rossella , Lolli Serena , Mattioli Claudia , Tampella Giacomo , Cattalini Marco , Kilic Sebnem S. , Pandolfi Franco , Plebani Alessandro , Frezza Domenico
Published: December 2009

Selective IgA deficiency (IGAD) is the most common primary immunodeficiency, yet its pathogenesis is elusive. The IG (heavy) H chain human 3′ Regulatory Region harbors three enhancers and has an important role in Ig synthesis. HS1.2 is the only polymorphic enhancer of the 3′RRs. We therefore evaluated HS1.2 allelic frequencies in 88 IGAD patients and 101 controls. Our data show that IGAD patients have a highly significant increase of homozygousity of the allele *1 (39% in the IGAD patients and 15% in controls), with an increase of 2.6-fold. Allele *4 has a similar trend of allele *2, both showing a significant decrease of frequency in IGAD. No relationship was observed between allele *1 frequencies and serum levels of IgG. However, allele *1 was associated in IGAD patients with relatively low IgM levels (within the 30th lowest percentile of patients). The HS1.2 polymorphism influences Ig seric production, but not IgG switch, in fact 30th lowest or highest percentile of IgG in patients did not associate to different frequencies of HS1.2 alleles. The control on normal healthy subjects did not correlate high or low levels of IgM or IgG with HS1.2 allelic frequence variation. Overall our candidate gene approach confirms that the study of polymorphisms in human diseases is a valid tool to investigate the function of these Regulatory Regions that confers multiple immune features.
Involvement of the NLRP3 Inflammasome in Innate and Humoral Adaptive Immune Responses to Fungal β-Glucan
Authors: Kumar Himanshu , Kumagai Yutaro , Tsuchida Tetsuo , Koenig Paul A. , Satoh Takashi , Guo Zijin , Jang Myoung Ho , Saitoh Tatsuya , Akira Shizuo , Kawai Taro
Published: December 2009

Fungal β-glucan, such as curdlan, triggers antifungal innate immune responses as well as shaping adaptive immune responses. In this study, we identified a key pathway that couples curdlan to immune responses. Curdlan promoted the production of the proinflammatory cytokine IL-1β by dendritic cells and macrophages through the NLRP3 inflammasome. Stimulation with Candida albicans and Saccharomyces cerevisiae also triggered the NLRP3 inflammasome-mediated IL-1β production. In vivo, NLRP3 was required for efficient Ag-specific Ab production when curdlan was used as an adjuvant, whereas it was dispensable for the induction of Th1 and Th17 cell differentiation. Furthermore, stimulation of purified B cells with curdlan-induced CD69 up-regulation and IgM production while stimulation with other NLRP3 inflammasome activators, such as silica and aluminum salt, did not. Notably, this induction required NLRP3 but was independent of Toll-like receptor and IL-1 receptor family signaling, suggesting the presence of NLRP3-dependent and IL-1 receptor family independent mechanisms in B cells responsible for Ab responses. Collectively, these findings reveal a critical role for the NLRP3 inflammasome in the regulation of antifungal innate immune responses as well as B cell activation.
Dynamics of the Interaction of Human IgG Subtype Immune Complexes with Cells Expressing R and H Allelic Forms of a Low-Affinity Fcγ Receptor CD32A
Authors: Shashidharamurthy Rangaiah , Zhang Fang , Amano Aaron , Kamat Aparna , Panchanathan Ravichandran , Ezekwudo Daniel , Zhu Cheng , Selvaraj Periasamy
Published: December 2009

CD32A, the major phagocytic FcγR in humans, exhibits a polymorphism in the ligand binding domain. Individuals homozygous for the R allelic form of CD32A (CD32AR allele) are more susceptible to bacterial infections and autoimmune diseases as compared with H allelic CD32A (CD32AH) homozygous and CD32AR/H heterozygous individuals. To understand the mechanisms behind this differential susceptibility, we have investigated the dynamics of the interaction of these allelic forms of CD32A when they are simultaneously exposed to immune complexes (IC). Binding studies using Ig fusion proteins of CD32A alleles showed that the R allele has significantly lower binding not only to human IgG2, but also to IgG1 and IgG3 subtypes. Competition assays using purified molecules demonstrated that CD32AH-Ig outcompetes CD32AR-Ig for IC binding when both alleles simultaneously compete for the same ligand. CD32AH-Ig blocked the IC binding mediated by both the allelic forms of cell surface CD32A, whereas CD32AR-Ig blocked only CD32AR and was unable to cross-block IC binding mediated by CD32AH. Two-dimensional affinity measurements also demonstrated that CD32AR has significantly lower affinity toward all three subtypes as compared with CD32AH. Our data suggest that the lower binding of CD32AR not only to IgG2 but also to IgG1 and IgG3 might be responsible for the lack of clearance of IC leading to increased susceptibility to bacterial infections and autoimmune diseases. Our data further suggests that in humans, inflammatory cells from CD32AR/H heterozygous individuals may predominantly use the H allele to mediate Ab-coated target cell binding during phagocytosis and Ab-dependent cellular cytotoxicity, resulting in a phenotype similar to CD32AH homozygous individuals.
Autoantibodies against C1q in Systemic Lupus Erythematosus Are Antigen-Driven
Authors: Schaller Monica , Bigler Cornelia , Danner Doris , Ditzel Henrik J. , Trendelenburg Marten
Published: December 2009

Autoantibodies against complement C1q (anti-C1q Abs) were shown to strongly correlate with the occurrence of severe nephritis in patients with systemic lupus erythematosus (SLE), suggesting a potential pathogenic role by interfering with the complement cascade. To analyze the humoral immune response against C1q at the molecular level, we screened a bone marrow-derived IgGκ/IgGλ Fab phage display library from a SLE patient with high anti-C1q Ab titer against purified human C1q. Six Fabs that exhibited strong binding to C1q in ELISA were isolated. The anti-C1q Fabs recognized neoepitopes that were only exposed on bound C1q and not present on soluble C1q mapping to different regions of the collagen-like region of C1q. Analysis of the genes encoding the variable H and L chains of the IgG-derived anti-C1q Fab revealed that all the variable H and L chain regions were highly mutated, with nucleotide and amino acid homologies to the closest germline in the range of 71–97% (average 85 ± 4) and 72–92% (average 88 ± 6), respectively. In addition, the variable region of the Fabs exhibited high replacement to silent ratios. The six anti-C1q Fabs were shown to be of high affinity, with a Kd ranging from of 8.4 × 10−8 M to 1.4 × 10−7 M, comparable to an antiviral immune response. Our data underlines the notion that the development of anti-C1q Abs in SLE is the consequence of an Ag-driven, affinity-matured immune response. Those anti-C1q Fabs are unique tools to address how complement C1q is implicated in the pathogenesis of SLE.
Differential Responses to Smith D Autoantigen by Mice with HLA-DR and HLA-DQ Transgenes: Dominant Responses by HLA-DR3 Transgenic Mice with Diversification of Autoantibodies to Small Nuclear Ribonucleoprotein, Double-Stranded DNA, and Nuclear Antigens
Authors: Jiang Chao , Deshmukh Umesh S. , Gaskin Felicia , Bagavant Harini , Hanson Julie , David Chella S. , Fu Shu Man
Published: December 2009

Anti-Smith (Sm) D autoantibodies are specific for systemic lupus erythematosus. In this investigation, the influence of HLA-D genes on immune responses to SmD was investigated. Mice with HLA-DR3, HLA-DR4, HLA-DQ0601, HLA-DQ0604, or HLA-DQ8 transgenes were immunized with recombinant SmD1, and their Ab responses were analyzed. Analysis by ELISA showed that all strains responded well to SmD. However, when synthetic SmD peptides were used as substrate, DR3 mice had the highest Ab response followed by DQ8, DQ0604, DQ0601, and DR4. A similar trend was observed in Western blot analysis using WEHI 7.1 cell lysate as the substrate, with the exception that DR4 mice did not generate detectable amounts of Abs. Only sera from DR3 and DQ0604 mice immunoprecipitated A-ribonucleoprotein (RNP), SmB, and SmD. Intermolecular epitope spreading to A-RNP and SmB was evident in DR3 and DQ0604 mice, as sera depleted of anti-SmD Abs were reactive with these proteins. DR3 mice also generated an immune response to C-RNP. Anti-nuclear Abs were detected in the majority of the DR3 mice, whereas moderate reactivities were seen in DQ0604 and DQ8 mice. Interestingly, only DR3 mice mounted an anti-dsDNA Ab response. Approximately half of the anti-dsDNA Abs were cross-reactive with SmD. Ab responses correlated with the strength of the T cell responses. Thus, HLA-DR3 appears to be the dominant HLA-D gene that determines the magnitude and quality of the anti-SmD immune response. In addition, our findings provide insights into the origin of the anti-dsDNA Abs often detected in patients with systemic lupus erythematosus.
Acute neurological problems: frequency, consultation patterns and the uses of a rapid access neurology clinic
Authors: Chapman FA , Pope AE , Sorensen D , Knight RSG , Al-Shahi Salman R
Published: December 2009

In secondary care, some patients with acute neurological symptoms are never seen by a neurologist. Rapid access neurology clinics could provide patients with timely access to neurology services. We analysed a retrospective cohort of 12,024 consecutive patients attending the ‘immediate care’ area of the emergency department or the acute medical admissions unit of the Royal Infirmary of Edinburgh. A total of 1,036 patients (9%) presented with a neurological complaint, of whom 680 (66%) did not have any contact with neurology services. The most common problems were epileptic seizure, cerebrovascular diseases and headache. Of the patients with epileptic seizure or headache who were not seen by a neurologist, about 40% might have benefited from neurological assessment. Following the introduction of a weekly rapid access neurology clinic, the most common problems seen were headache, symptoms that turned out to be medically unexplained and epileptic seizure.
Serum potassium, calcium and magnesium in patients receiving ESHAP chemotherapy for relapsed lymphomas
Authors: Zekri J , Cheah NLC , Evans L , Hancock B
Published: December 2009

Etoposide, methylprednisolone, cytarabine and cisplatin (ESHAP) is one of the mostly widely used chemotherapy regimens for patients with relapsed lymphomas. Cisplatin administration is commonly associated with electrolyte imbalance. Careful monitoring of renal function and serum electrolytes is therefore essential in this setting.

Aims: To review the practice of electrolyte monitoring – potassium (K), calcium (Ca) and magnesium (Mg) – in patients receiving ESHAP and the frequency and severity of abnormalities and their management.

Methods: Twenty-one consecutive patients received ESHAP. The medical records of 16 patients were retrievable and reviewed retrospectively. Results of serum K, Ca and Mg were collected prior to and after cycles 1, 2 and 3 of ESHAP, if measured.

Results: Serum K levels prior to every cycle did not show any noticeable change. The means were 4.42, 4.34 and 4.43 mmol/l before cycles 1, 2 and 3, respectively. In one patient hypokalaemia was severe, refractory and symptomatic when preceded by hypomagnesaemia. Serum-adjusted calcium levels showed only minimal reduction. The means were 2.46, 2.40 and 2.38 mmol/l before cycles 1, 2 and 3 respectively. Mean serum Mg levels prior to every cycle showed progressive reduction; 0.84, 0.75 and 0.67 mmol/l before cycles 1, 2 and 3, respectively. This was associated with a progressive increase in the amount of required Mg supplementation. Serum K, Ca and Mg was measured prior to 100%, 67% and 35% of administered cycles, respectively.

Conclusion: In patients receiving ESHAP, hypokalaemia can occasionally be seen, especially if preceded by hypomagnesaemia. Mild cumulative hypocalcaemia is recognised. Hypomagnesaemia is a progressive complication and physicians need be aware of its importance.
Neuropsychiatric features of Parkinson’s disease
Authors: Turnbull C , Fitzsimmons P
Published: December 2009

Parkinson’s disease is a chronic, progressive, neurodegenerative condition that damages central nervous system pathways. In addition to the wellrecognised motor features of the disease, patients commonly experience nonmotor symptoms including neuropsychiatric complications. These symptoms may precede motor symptoms and have a very significant impact on quality of life for patients and their carers.
Hormone replacement therapy: the risks and benefits of treatment
Authors: Hardie C , Bain C , Walters M
Published: December 2009

Several recent large studies have provoked concern among both health professionals and the general public regarding the safety of hormone replacement therapy (HRT). This article provides a review of the current literature surrounding the risks and benefits of HRT in postmenopausal women, and how the data can be applied safely in everyday clinical practice.
The future of genetics in everyday medicine
Authors: Thalayasingam N
Published: December 2009

Genetics is a developing field that increasingly influences medical practice in a large number of specialties. This symposium highlighted how current research is providing insights into the genetic causes of disease, leading to a deeper understanding of the pathophysiology and identification of potential novel treatment strategies. We were provided with an overview of developments in monogenic conditions such as Marfan’s syndrome and Duchenne muscular dystrophy and complex traits such as atopic dermatitis. The practicalities of providing genetic testing for more common conditions such as familial hypercholesterolaemia were discussed, highlighting the more prominent role clinical genetics may come to play in mainstream medicine.
Care of the elderly symposium report
Authors: McGeough EC
Published: December 2009

This symposium covered a wide range of conditions of interest to the geriatrician, the general physician and the general practitioner, including demographic shift, the epidemiology of ageing, diabetes in older people, investigation and management of falls, an update on stroke (including the role of neurovascular clinics, stroke thrombolysis and rehabilitation) and the management of coronary heart disease in the elderly.
Nonenzymatic Oligomerization of Activated Nucleotides on Hairpin Templates
Authors: Kim Eun‐Kyong , Switzer Christopher
Published: December 2009

This unit describes a protocol for nonenzymatic oligomerization of activated ribonucleotides on DNA hairpins appended by templates containing threofuranosyl nucleic acid (TNA). TNA‐cytidylate templates effectively promote oligomerization of 2‐MeImpG, and give 3′,5′‐linked oligomerization products predominantly, with good base‐pairing fidelity. Although the rates of oligomerization depend on TNA content, after 3 days of incubation, oligomerization products are apparent, and full‐length products are present after 10 days. Characterization of product phosphodiester bond regiochemistry is accomplished by digestion with RNase T1. Additionally, exposure of oligomerization products to calf intestinal alkaline phosphatase enables detection of any endcapping due to pyrophosphate formation. Base‐pairing fidelity is assessed by challenging the template to oligomerize 2‐MeImpA. The protocols described for nonenzymatic, template‐directed synthesis in this unit are applicable to oligomerization of activated monomers on templates of different compositions, with respect to both base identity and polymer backbone. Curr. Protoc. Nucleic Acid Chem. 39:3.18.1‐3.18.13. © 2009 by John Wiley & Sons, Inc.
Fluorescence Correlation Spectroscopy in Living Cells: A Practical Approach
Authors: Altan‐Bonnet Nihal , Altan‐Bonnet Grégoire
Published: December 2009

Fluorescence correlation spectroscopy (FCS) is a single‐molecule fluorescence technique used to monitor molecular dynamics. It can be applied to living cells expressing fluorescently labeled proteins and lipids to determine the diffusion timescales and the total number of diffusing fluorescent molecules in the cell. A practical step‐by‐step approach to performing FCS with a commercial spectroscopy/microscopy system, the Zeiss Confocor 3, how to set up live‐cell FCS experiments, acquire reliable data, and finally how to analyze the data acquired, are all described in this unit. Curr. Protoc. Cell Biol. 45:4.24.1‐4.24.14. © 2009 by John Wiley & Sons, Inc.
Generation of Micropatterned Substrates Using Micro Photopatterning
Authors: Doyle Andrew D.
Published: December 2009

Micro photopatterning (µPP) has been developed to rapidly test and generate different patterns for extracellular matrix adsorption without being hindered by the process of making physical stamps through nanolithography techniques. It uses two‐photon excitation guided through a point‐scanning confocal microscope to locally photoablate poly(vinyl) alcohol (PVA) thin films in user‐defined computer‐controlled patterns. PVA thin films are ideal for surface blocking, being hydrophilic substrates that deter protein adsorption and cell attachment. Because gold substrates are not used during µPP, all live‐cell fluorescent‐imaging techniques including total internal reflection fluorescence microscopy of GFP–linked proteins can be performed with minimal loss of fluorescence signal. Furthermore, because µPP does not require physical stamps for pattern generation, multiple ECMs or other proteins can be localized within microns of each other. This unit details the setup of µPP. It also provides troubleshooting techniques. Curr. Protoc. Cell Biol. 45:10.15.1‐10.15.35. © 2009 by John Wiley & Sons, Inc.
Microstructural behaviors of polyaniline/CB Composites by SAXS
Authors: Siddaramaiah  , Souza Fernando G. , Soares Bluma G. , Parameswara P. , Somashekar R.
Published: December 2009

Polyaniline doped with dodecylbenzene sulfonic acid and conductive carbon black (PAn.DBSA/CB) hybrid materials have been prepared by in situ polymerization. The electrical resistivity of the PAn/CB was measured as a function of CB. The minimum resistivity was noticed for the composites with 25 wt % of CB as compared to other systems. Small angle x‐ray diffraction data was used to characterize the molecular arrangements of these composites. The refinement parameters such as periodicity (L), phase lengths ( and ), phase ratio (/), and probability distribution of phases [γZ (crystalline), γY (amorphous)] for PAn, CB, and PAn/CB composites have been measured by SAXS data. It was observed that there is decrease in the crystallinity with increase in CB content in the composites. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Synthesis and characterization of polyaniline/silica doped with camphorsulfonic acid and dodecylbenzylsulfonic acid
Authors: Lee Hsun‐Tsing , Yang Shung‐Jim
Published: December 2009

In this study, the effect of the dopant structure on the morphology, structure, optical absorption, conductivity, and thermal properties of conductive polyaniline (PAn)/silica (SiO2) hybrids from a sol–gel process was first investigated. Camphorsulfonic acid (CSA) and dodecylbenzylsulfonic acid (DBSA) were used as the model acid dopants. The polaron absorption peaks of the CSA‐doped PAn/SiO2 (CSA–PAn/SiO2) hybrids were redshifted compared with those of the DBSA–PAn/SiO2 hybrids. This indicated that the doped PAn subchains of the CSA–PAn/SiO2 hybrids had a more rigid conformation and longer conjugation length than those of DBSA–PAn/SiO2 hybrids. Therefore, the CSA–PAn/SiO2 hybrids were more conductive than the DBSA–PAn/SiO2 hybrids with same SiO2 contents. Moreover, the CSA–PAn/SiO2 hybrids exhibited higher SiO2 condensation degrees and thermal resistance than the DBSA–PAn/SiO2 hybrids with same SiO2 contents. In addition, the CSA–PAn/SiO2 films exhibited a nonuniform globular morphology, whereas the DBSA–PAn/SiO2 films showed an interconnected and rugged fibrillar one. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010
Semantic annotation of biosystematics literature without training examples
Authors: Cui Hong , Boufford David , Selden Paul
Published: December 2009

This article presents an unsupervised algorithm for semantic annotation of morphological descriptions of whole organisms. The algorithm is able to annotate plain text descriptions with high accuracy at the clause level by exploiting the corpus itself. In other words, the algorithm does not need lexicons, syntactic parsers, training examples, or annotation templates. The evaluation on two real‐life description collections in botany and paleontology shows that the algorithm has the following desirable features: (a) reduces/eliminates manual labor required to compile dictionaries and prepare source documents; (b) improves annotation coverage: the algorithm annotates what appears in documents and is not limited by predefined and often incomplete templates; (c) learns clean and reusable concepts: the algorithm learns organ names and character states that can be used to construct reusable domain lexicons, as opposed to collection‐dependent patterns whose applicability is often limited to a particular collection; (d) insensitive to collection size; and (e) runs in linear time with respect to the number of clauses to be annotated.
Analysis of phenolic compounds in health care products by low‐pressure liquid‐chromatography with monolithic column and chemiluminescent detection
Authors: Ballesta‐Claver J. , Valencia M. C. , Capitán‐Vallvey L. F.
Published: December 2009

This paper presents a new application for monolithic columns with low‐pressure chromatographic separation using an flow injection analysis configuration with chemiluminescent detection for the determination of a mixture of phenolic compounds: phloroglucinol, 2,4‐dihydroxybenzoic acid, salicylic acid, methyl paraben and n‐propyl gallate. The procedure consists of the separation of these compounds on a reverse‐phase ultra‐short monolithic column with pH 3.0 acetate buffer and 5% acetonitrile as carrier phase. The detection is based on a chemiluminescence measurement coming from Ce(IV)–Rhodamine 6G chemistry with the incorporation of two different chemiluminescent chemical conditions in the chromatographic setup in order to enhance the sensitivity for the different phenolic compounds. All separation and detection variables were optimized to propose a determination method. The analysis is performed in 280?s, with the sampling frequency being some 13 h−1. The calibration function is a double reciprocal function obtaining good results within two orders of magnitude. The limits of detection were 8.8 × 10 −8 m (phloroglucinol), 2.7 × 10 −8 m (2,4‐dihydroxybenzoic acid); 2.3 × 10 −8 m (salicylic acid); 5.2 × 10 −8 m (methyl paraben) and 4.1 × 10 −6 m (n‐propyl gallate), and the relative standard deviations at a medium level of the linear range were 4.4% (phloroglucinol), 2.8% (2,4‐dihydroxybenzoic acid), 5.2% (salicylic acid), 3.6% (methyl paraben) and 6.8% (n‐propyl gallate). The method was applied and validated satisfactorily for the determination of these compounds in healthcare products, comparing the results against an HPLC reference method. Copyright © 2009 John Wiley & Sons, Ltd.
Meta-analysis of randomized controlled trials comparing laparoscopic with open mesh repair of recurrent inguinal hernia
Authors: Karthikesalingam A , Markar S R , Holt P J E , Praseedom R K
Published: December 2009

Background

Although there is plentiful evidence regarding the use of laparoscopic surgery for primary inguinal hernia, there is a paucity of literature concerning its role after recurrence. There has been no quantitative review of the evidence, despite suggestions that pooled analysis of existing data is required.

Methods

Medline, Embase, trial registries, conference proceedings and reference lists were searched for controlled trials of laparoscopic versus conventional open surgery for mesh repair of recurrent hernia. The primary outcomes were recurrence and chronic pain. Secondary outcomes were operating time, visual analogue pain score, superficial wound infection, haematoma or seroma formation, time to return to normal activities and serious complications requiring operation. Pooled odds ratios were calculated for categorical outcomes and weighted mean differences for continuous outcomes.

Results

Four trials were included in the analysis. There was no effect on recurrence or chronic pain. Laparoscopic surgery was associated with significantly less postoperative pain, a quicker return to normal activities and fewer wound infections, at the cost of a longer operating time. There was no difference in haematoma formation or the need for additional operations.

Conclusion

Careful patient selection and surgeons' experience are important in the selection of technique for recurrent inguinal hernia repair.
Influence of Gold Nanoparticles of Varying Size in Improving the Lipase Activity within Cationic Reverse Micelles
Authors: Maiti Subhabrata , Das Dibyendu , Shome Anshupriya , Das Prasanta Kumar
Published: December 2009

Herein, we report the effect of gold nanoparticles (GNPs) in enhancing lipase activity in reverse micelles of cetyltrimethylammonium bromide (CTAB)/water/isooctane/n‐hexanol. The size and concentration of the nanoparticles were varied and their specific roles were assessed in detail. An overall enhancement of activity was observed in the GNP‐doped CTAB reverse micelles. The improvement in activity becomes more prominent with increasing concentration and size of the GNPs (0–52 μM and ca. 3–30 nm, respectively). The observed highest lipase activity (k2=1070±12 cm3 g−1 s−1) in GNP‐doped CTAB reverse micelles ([GNP]: 52 μm, ca. 20 nm) is 2.5‐fold higher than in CTAB reverse micelles without GNPs. Improvement in the lipase activity is only specific to the GNP‐doped reverse micellar media, whereas GNP deactivates and structurally deforms the enzyme in aqueous media. The reason for this activation is probably due to the formation of larger‐sized reverse micelles in which the GNP acts as a polar core and the surfactants aggregate around the nanoparticle (‘GNP pool’) instead of only water. Lipase at the augmented interface of the GNP‐doped reverse micelle showed improved activity because of enhancement in both the substrate and enzyme concentrations and increased flexibility in the lipase conformation. The extent of the activation is greater in the case of the larger‐sized GNPs. A correlation has been established between the activity of lipase and its secondary structure by using circular dichroism and FTIR spectroscopic analysis. The generalized influence of GNP is verified in the reverse micelles of another surfactant, namely, cetyltripropylammonium bromide (CTPAB). TEM, dynamic light scattering (DLS), and UV/Vis spectroscopic analysis were utilized to characterize the GNPs and the organized aggregates. For the first time, CTAB‐based reverse micelles have been found to be an excellent host for lipase simply by doping with appropriately sized GNPs.
Chiral Auxiliaries as Emerging Tools for the Asymmetric Synthesis of Octahedral Metal Complexes
Authors: Meggers Eric
Published: December 2009

New methods for the stereocontrolled synthesis of octahedral metal complexes are needed in order to fully exploit the stereochemical richness of the octahedron in the fields of catalysis, materials sciences, and life sciences. Whereas a large body of work exists regarding the diastereoselective coordination chemistry with chiral ligands, such efforts are restricted to certain carefully designed chiral ligands which remain in the coordination sphere. The emerging strategy of chiral‐auxiliary‐mediated asymmetric synthesis holds promise to solve the problem of controlling relative and absolute configuration in octahedral metal complexes in a general fashion, thus hopefully in the future providing access to any desired optical active octahedral metal complex without the need for chiral separations. This short review will summarize reported examples of chiral auxiliaries applied to the asymmetric synthesis of octahedral metal complexes.
CB1 receptor inverse agonist pharmacotherapy for metabolic disorders
Authors: Fong Tung M. , Addy Carol , Erondu Ngozi , Heymsfield Steven B.
Published: December 2009

The cannabinoid‐1 receptor (CB1R) has been implicated in the control of energy balance based on its anatomical localization, the interaction among the endocannabinoid system and other transmitter systems, and the appetite‐enhancing effect of tetrahydrocannabinol. Recent development of CB1R inverse agonists indicated that these agents inhibit food intake, increase energy expenditure, and reduce fat mass. This article reviews the preclinical and clinical efficacy of several CB1R inverse agonists and provides a perspective on the use of CB1R inverse agonists in studies of obesity and metabolic disorders. The compounds reviewed here are no longer in clinical development for obesity. Drug Dev Res 70:566–576, 2009. © 2009 Wiley‐Liss, Inc.
Recent CB1 cannabinoid receptor antagonists and inverse agonists
Authors: Seltzman Herbert H.
Published: December 2009

Antagonists or inverse agonists of the cannabinoid CB1 receptor (CB1R) reported from the 2006–2007 period are reviewed. The compounds are either variations of the archetypical cannabinoid CB1 inverse agonist rimonabant (SR141716) or are other structures less obviously related to the archetype. Many of the former class replace the core pyrazole ring of rimonabant with other ring systems to display/arrange the appended binding/activity elements. A structure‐activity relationship evolves from bioassays of these compounds that also provides a more detailed understanding of the molecular mechanism of the binding to and stabilization of the active and inactive states of the constitutively active CB1R. In addition to diverse variations of the core ring and appended substituents, conformationally constrained analogs with high affinity were revealed, a number of which do not fit a one‐to‐one atom correspondence with the putatively active conformations of the rimonabant template and other active analogs. Diphenylmethyl analogs were a repeated motif in structures where core rings were either absent or played less restrictive roles in structures pursuing higher conformational freedom. Bicyclic nitrogen heterocyclic ring systems as the central core structures were reported, as were linear structures including phenyl ureas and taranabant analogs. Finally, peptides in the hemopressin class were presented as CB1 antagonists. Drug Dev Res 70:601–615, 2009. © 2009 Wiley‐Liss, Inc.
Analysis of non‐ketoic steroids 17α‐methylepithiostanol and desoxymethyl‐ testosterone in dietary supplements
Authors: Okano Masato , Sato Mitsuhiko , Ikekita Ayako , Kageyama Shinji
Published: December 2009

Dietary supplements containing 17α‐methyl‐2,3‐epithio‐5α‐androstane‐17β‐ol (17α‐methylepithiostanol), which is a 17‐methylated analogue of epithiostanol or a prodrug of desoxymethyltestosterone (17α‐methyl‐5α‐androst‐2‐en‐17β‐ol), have recently appeared on the Internet. 17α‐Methylepithiostanol and desoxymethyltestosterone are classified as prohibited substances on the World Anti‐Doping Agency (WADA) list. Two preparations, EPISTANE™ and P‐PLEX™, were obtained from the Internet so that their contents could be investigated. This study involved gas chromatography/mass spectrometry (GC/MS) analysis after trimethylsilyl (TMS) derivatization, liquid chromatography/mass spectrometry (LC/MS) in atmospheric pressure photoionization (APPI) mode and nuclear magnetic resonance (NMR) spectroscopy. Analysis using LC/MS in APPI mode would be a useful tool for detecting heat‐labile and non‐polar steroids.

Although the labelling of EPISTANE™ indicates that it contains 17α‐methyl‐2α, 3α‐epithio‐5α‐androstane‐17β‐ol only, 17α‐methyl‐2β,3β‐epithio‐5α‐androstane‐17β‐ol and desoxymethyltestosterone were identified in the supplement. The results showed that P‐PLEX™ contained desoxymethyltestosterone and its isomer 17α‐methyl‐5α‐androst‐3‐en‐17β‐ol. Urine samples can be screened after EPISTANE™ or P‐PLEX™ administration using the normal screening procedure for anabolic steroids with GC/MS. Copyright © 2009 John Wiley & Sons, Ltd.
Coupled electrothermal analysis of a micro flow sensor with control circuit using spice
Authors: Kamiunten Shoji , Nagayo Hidetomo , Motosuke Masahiro , Honami Shinji
Published: December 2009

This paper reports an electrothermal analysis of a two‐element micro flow sensor (MFS). The measurement principle is based on temperature difference between the upstream and downstream parts of electrically connected heating‐sensing elements. The two‐heater MFS, the thermal fluid flow around MFS, and the electric circuit were modeled together using a commercial general‐purpose circuit simulator based on SPICE (Simulation Program with Integrated Circuit Emphasis). The model has a partial three‐dimensional lumped thermal network model involved in a two‐dimensional one. Two‐dimensional laminar Poiseuille channel flow was assumed as the flow field in the model, and the forced convective heat transfer was calculated by energy balance at each node in the fluid flow. The tendency on the sensor characteristics was in good agreement between the simulation and experiment. © 2009 Wiley Periodicals, Inc. Electron Comm Jpn, 93(1): 58–64, 2010; Published online in Wiley InterScience (www.interscience. wiley.com). DOI 10.1002/ecj.10095