bluuebunny/crossref_metadata_2025 huggingface dataset:
True prevalence of twin HDV-HBV infection in Pakistan: a molecular approach
Authors: Khan Asad U , Waqar Muhammad , Akram Madiha , Zaib Mehnaz , Wasim Muhammad , Ahmad Shahzad , Niaz Zeeshan , Ali Sajid , Ali Haider , Idrees Muhammad , Bajwa Mohammad A
Published: September 2011

Hepatitis Delta Virus (HDV) infects only patients that are already infected by hepatitis B virus (HBV) because this is sub satellite virus which depends on and propagate only in the presence of HBV. HDV causes co-infection or super infection with sever complication as compared to only HBV infection. No study on molecular level on HDV is available from this region; therefore, the aim of this study was to found out the molecular epidemiology of HDV (as a co-infection with HBV) in different geographical regions of Pakistan.

Total 228 HBsAg positive samples were received for the study from different geographical regions of the country. Only HBV DNA PCR positive samples were further utilized for the presence of HDV RNA. For this purpose, HDV RNA and HBV DNA was extracted and amplified using reverse transcriptase polymerase chain reaction (RT-PCR), nested PCR and real-time PCR.

Out of the total 228 HBsAg positive samples, HBV DNA was detected in total 190 (83.3%) samples belonged to different patients. Of these 190 patients, HDV RNA was observed in 53 (28%) patients. Of the 53 HDV positive cases, 37 (69.8%) were males and 16 (30.2%) were female patients. The percentage of dual infection was found higher significantly (p < 0.05) in male patients as compared to female patients. Total 41 (26.8%) patients were below 40 years and 13 (31.7%) were above 40 years of age. No significant difference was seen in patients with ages above or below 40 years. In the provinces of Sindh, Khyber Pakhtoonkhaw and Punjab the observed prevalence of HDV was 67%, 6% and 4% respectively.

In conclusion, the HDV infection is not uncommon in Pakistan and its prevalence is higher significantly in the Province of Sindh (p < 0.01) and male six (p < 0.05).
XMRV: usage of receptors and potential co-receptors
Authors: Haleyur Giri Setty Mohan Kumar , Devadas Krishnakumar , Ragupathy Viswanath , Ravichandran Veerasamy , Tang Shixing , Wood Owen , Gaddam Durga Sivacharan , Lee Sherwin , Hewlett Indira K
Published: September 2011

Background

XMRV is a gammaretrovirus first identified in prostate tissues of Prostate Cancer (PC) patients and later in the blood cells of patients with Chronic Fatigue Syndrome (CFS). Although XMRV is thought to use XPR1 for cell entry, it infects A549 cells that do not express XPR1, suggesting usage of other receptors or co-receptors.

Methods

To study the usage of different receptors and co- receptors that could play a role in XMRV infection of lymphoid cells and GHOST (GFP- Human osteosarcoma) cells expressing CD4 along with different chemokine receptors including CCR1, CCR2, etc., were infected with XMRV. Culture supernatants and cells were tested for XMRV replication using real time quantitative PCR.

Results

Infection and replication of XMRV was seen in a variety of GHOST cells, LNCaP, DU145, A549 and Caski cell lines. The levels of XMRV replication varied in different cell lines showing differential replication in different cell lines. However, replication in A549 which lacks XPR1 expression was relatively higher than DU145 but lower than, LNCaP. XMRV replication varied in GHOST cell lines expressing CD4 and each of the co- receptors CCR1-CCR8 and bob. There was significant replication of XMRV in CCR3 and Bonzo although it is much lower when compared to DU145, A549 and LNCaP.

Conclusion

XMRV replication was observed in GHOST cells that express CD4 and each of the chemokine receptors ranging from CCR1- CCR8 and BOB suggesting that infectivity in hematopoietic cells could be mediated by use of these receptors.
Adenoid cystic carcinoma intermingled with ductal carcinoma of the breast: a case report and review of the literature
Authors: Kontos Michael , Karles Dimitrios , Petrou Athanasios , Alexandrou Paraskevi TH
Published: September 2011

Introduction

Adenoid cystic cancer of the breast is a rare condition, and even rarer are the cases where it is histologically mixed with other variants of cancer within a single lesion. In this report, one of the few cases of mixed adenoid cystic breast cancer intermingled with the infiltrating ductal variant is presented. A subsequent review of the relevant literature presents the existing experience in treating mixed breast cancers with adenoid cystic components with regard to diagnosis, treatment, and prognosis.

Case presentation

We describe a case of mixed adenoid cystic cancer of the breast with infiltrating ductal carcinoma in a 67-year-old Caucasian woman who underwent mastectomy with sentinel node biopsy.

Conclusion

Surgery remains the cornerstone of treatment of these patients, and radiotherapy is administered when breast-conserving treatment is undertaken or a large tumor with affected lymph nodes is present. Hormonal treatment does not have a role, as estrogen receptors are always absent from both tumor components. Chemotherapy is nearly always administered on the basis of estrogen receptor and progesterone negativity and the more aggressive potential of the non-adenoid cystic component. The de-differentiation of an indolent type of cancer to a more aggressive one may affect the prognosis.
”I could cry, the amount of shoes I can't get into”:A qualitative exploration of the factors that influence retail footwear selection in women with rheumatoid arthritis
Authors: Naidoo Serena , Anderson Stephanie , Mills Joanna , Parsons Stephanie , Breeden Stephanie , Bevan Emma , Edwards Camilla , Otter Simon
Published: September 2011

Background

Studies have reported that women with rheumatoid arthritis (RA) are not wearing NHS supplied therapeutic footwear; therefore it is likely they are wearing footwear sourced through retailers. Previous research gives limited information (largely associated with cosmesis) on people's perceptions on the relationships that exist between retail footwear, well‐being and quality of life. This study aimed to explore the perceptions of women with RA regarding their choice of retail footwear and identify the factors influencing retail footwear selection.

Methods

Eleven women with RA wearing normal retail footwear were recruited from an out‐patient podiatry clinic in the south east of England. Semi‐structured interviews were carried out and an interpretative phenomenological approach was adopted for data collection and transcript analysis.

Results

Six key themes were revealed from the analysis: (1) the nature of foot complaints and deformities, (2) aesthetic appearance and design of footwear, (3) body image, (4) psychosocial aspects, (5) Perceptions of footwear and (6) the therapeutic value of retail shoes. These contributed to an overarching concept of loss of choice associated with retail footwear. In particular, the areas discussed most frequently throughout were themes (2), (3) and (4), which were notably more ‘emotional’ in nature.

Conclusions

Limitations in retail footwear for these women have impacted on their individuality, linking significantly with their body image. The loss of choice in footwear as a consequence of the disease impacts negatively on emotions, wellbeing and was identified in reduced self‐perceived quality of life.
[3 + 2] Cycloaddition reactions of thioisatin with thiazolidine-2-carboxylic acid: a versatile route to new heterocyclic scaffolds
Authors: Verma Sonali , George Johnson , Singh Saurabh , Pardasani Pushpa , Pardasani Ramchand
Published: September 2011

A facile synthesis of azabicycloadducts is described by 1,3-dipolar cycloaddition reactions of thioisatin with thiazolidine-2-carboxylic acid in the presence of various electron rich and electron deficient dipolarophiles. Theoritical calculations have been performed to study the regioselectivity of products. The geometrical and energetic properties have been analyzed for the different reactants, transition states and cycloadducts formed.
Macroeconomics and Methodology
Authors: Sims Christopher A
Published: September 2011

Probabilistic reasoning is essential to discourse in economics. This is true in any discipline in which, as in economics, data collection is constrained and beliefs about the phenomena being studied are crucial to decisions that cannot be delayed. Some economists have recently turned away from form probabilistic inference, in part because of legitimate discontent with the prescriptions of specialized econometricians. However, this turning away has gone in mutually inconsistent directions and is in the long run unsustainable. It should be more widely recognized that careful applied work in macroeconomics, using steadily advancing probabilistic modeling techniques, has been steadily increasing.
The Empirical Foundations of Calibration
Authors: Hansen Lars Peter , Heckman James J
Published: September 2011

Interest in simulating recently developed dynamic stochastic general equilibrium models of the economy stimulated a demand for parameters. This has given rise to calibration as advocated by Finn E. Kydland and Edward C. Prescott (1982). This paper explores the implicit assumptions underlying their calibration method. The authors question that there is a ready supply of micro estimates available to calibrate macroeconomic models. Measures of parameter uncertainty and specification sensitivity should be routinely reported. They propose a more symbiotic role for calibration as providing signals to microeconomists about important gaps in knowledge, which when filled will solidify the empirical underpinning, improving the credibility of the quantitative output.
Distinguished Lecture on Economics in Government: Big Bills Left on the Sidewalk: Why Some Nations are Rich, and Others Poor
Authors: Olson Mancur
Published: September 2011

Some research presumes that, when rational parties bargain, nothing is left on the table, so that social outcomes are efficient and leave countries on the frontiers of their aggregate production functions. A study of differences in per capita incomes across countries shows that this cannot be the case. Countries’ endowments of natural and human resources do not explain any significant part of the variation in incomes and the mobility of capital assures that it is impartially available to all countries. National differences in the quality of policies and institutions across countries mainly account for differences in per capita incomes.
Privatization Is Transition—Or Is It?
Authors: Brada Josef C
Published: September 2011

Various means of creating a private sector have been used in transition economies, including restitution, privatization of state firms through sales or voucher schemes, and start-ups of new private firms. The privatization of state-owned firms raises questions regarding the viability of future corporate governance but, in combination with economic liberalization, it has induced state-owned firms to react to their new environment. Differences in the extent of privatization among countries reflect largely differences in the privatization of state-owned firms. Qualitative differences reflect differences in who the new owners of these firms are.
Policy Watch: Medicare
Authors: Newhouse Joseph P
Published: September 2011

The forecast growth for Medicare spending has created a highly visible budgetary impasse between the president and Congress. Both favor the growth of health plans that accept risk and would promote them by creating less restrictive options than heretofore. Nonetheless, the conference bill the president vetoed for other reasons did not envision price competition among plans but relied upon administered prices instead. The bill allowed Medical Savings Accounts; because of selection, the Congressional Budget Office estimated they would slightly increase Medicare costs. By eliminating the subsidy at the margin to hiring a resident, the bill's provisions would shift demand for residents down.
Data Watch: The Redesigned Current Population Survey
Authors: Polivka Anne E
Published: September 2011

The Current Population Survey (CPS), a national survey of 50,000 households, is a major source of information about the American labor market. In January 1994, the CPS underwent a major redesign both in the wording of the questionnaire and the methodology used to collect the data. This article reviews the motivation for the redesign, compares several key CPS estimates before and after the implementation of the new survey, and explains some of the new data collected in the redesigned CPS.
Motor Chip: A Comparative Genomic Hybridization Microarray for Copy-Number Mutations in 245 Neuromuscular Disorders
Authors: Piluso Giulio , Dionisi Manuela , Del Vecchio Blanco Francesca , Torella Annalaura , Aurino Stefania , Savarese Marco , Giugliano Teresa , Bertini Enrico , Terracciano Alessandra , Vainzof Mariz , Criscuolo Chiara , Politano Luisa , Casali Carlo , Santorelli Filippo Maria , Nigro Vincenzo
Published: September 2011

BACKGROUND

Array-based comparative genomic hybridization (aCGH) is a reference high-throughput technology for detecting large pathogenic or polymorphic copy-number variations in the human genome; however, a number of quantitative monogenic mutations, such as smaller heterozygous deletions or duplications, are usually missed in most disease genes when proper multiplex ligation-dependent probe assays are not performed.

METHODS

We developed the Motor Chip, a customized CGH array with exonic coverage of 245 genes involved in neuromuscular disorders (NMDs), as well as 180 candidate disease genes. We analyzed DNA samples from 26 patients with known deletions or duplications in NMDs, 11 patients with partial molecular diagnoses, and 19 patients with a clinical diagnosis alone.

RESULTS

The Motor Chip efficiently confirmed and refined the copy-number mutations in all of the characterized patients, even when only a single exon was involved. In noncharacterized or partially characterized patients, we found deletions in the SETX (senataxin), SGCG [sarcoglycan, gamma (35kDa dystrophin-associated glycoprotein)], and LAMA2 (laminin, alpha 2) genes, as well as duplications involving LAMA2 and the DYSF [dysferlin, limb girdle muscular dystrophy 2B (autosomal recessive)] locus.

CONCLUSIONS

The combination of exon-specific gene coverage and optimized platform and probe selection makes the Motor Chip a complementary tool for molecular diagnosis and gene investigation in neuromuscular diseases.
Attitudes toward depression among a sample of Muslim adolescents in the Midwestern United States
Authors: Haroun Zubeir , Bokhari Ali , Marko-Holguin Monika , Blomeke Kelsey , Goenka Ajay , Fogel Joshua , Van Voorhees Benjamin W.
Published: September 2011

Background: Little is known about how Muslim youth in non-Muslim countries perceive depression and its treatment and prevention.

Objective: We investigated the barriers and suggest treatment models for depressive disorders in Muslim adolescents and young adults residing in the United States.

Methods: We conducted a thorough literature review to identify previous study on the beliefs of American Muslim adolescents about depression and its treatment. We identified the gaps and developed a survey to ascertain this information from American Muslim adolescents.

Results: The survey was administered to a convenience sample of 125 Muslim subjects (60.0% males) aged 14–21 years. The sample was ethnically diverse with Pakistani (44.8%) encompassing the majority of the sample. Most responders believed that recitation from the Koran relieves mental distress. Multiple linear regression analysis revealed that those who reported strong emotional support from parents or a greater acceptance of taking depression medication prescribed from a physician were more likely to accept a physician’s diagnosis, whereas believing in prayer to heal depression was associated with a lower likelihood of the same. Youth were accepting of Internet and preventive approaches.

Conclusion: Planning of culturally sensitive mental health services is useful to accommodate the needs of Muslim youth served by primary care physicians and mental health practitioners in the United States. Muslim adolescents tend to be more traditional with family, social, and religious values. This value system plays an important role in their likelihood of seeking and accepting professional help for depression.
Conductometric study of sodium dodecyl sulfate - nonionic surfactant (Triton X-100, Tween 20, Tween 60, Tween 80 or Tween 85) mixed micelles in aqueous solution
Authors: Cirin Dejan , Posa Mihalj , Krstonosic Veljko , Milanovic Maja
Published: September 2011

The present study is concerned with the determination of the critical micelle
concentration (cmc) of mixed micelles of sodium dodecyl sulfate with one of
five nonionic surfactants (Triton X-100, Tween 20, Tween 60, Tween 80 or
Tween 85) from conductance measurements. Based on the calculated values of
the ? parameters we have noticed that SDS-nonionic surfactants mostly showed
strong synergistic effect. It was found that nonionic surfactants with
mainly longer and more hydrophobic tail show stronger interactions with
hydrophobic part of SDS, thus expressing stronger synergism. In SDS-Tween 80
binary system the strongest synergistic effect was noticed. SDS-Tween 85
micellar system showed antagonistic effect, most probably because the
presence of the double bond in its three hydrophobic tails (three C18 tails)
makes it sterically rigid.
Fabrication of multi-parametric platforms based on nanocone arrays for determination of cellular response
Authors: Purwaningsih Lindarti , Schoen Tobias , Wolfram Tobias , Pacholski Claudia , Spatz Joachim P
Published: September 2011

Cellular response to both surface topography and surface chemistry has been studied for several years. However, most of the studies focus on only one of the two parameters and do not consider their possible synergistic effects. Here, we report on a fabrication method for nanostructured surfaces composed of highly ordered arrays of silica nanocones with gold tips. By using a combination of block copolymer nanolithography, electroless deposition, and reactive ion etching several parameters such as structure height and structure distance could easily be adjusted to the desired values. The gold tips allow for easy functionalization of the substrates through a thiol linker system. Improved neural cell adhesion can be obtained and is dependent on the nature of the nanocone surface, thus illustrating the influence of different surface topographies on the nanometer length scale, on a complex cellular behavior such as cell adhesion. Substrate and surface functionality are shown to last over several days, leading to the conclusion that the features of our substrates can also be used for longer term experiments. Finally, initial neural cell adhesion is found to be more prominent on substrates with short intercone distances, which is an important finding for research dealing with the reactions of neuron-like tissue in the immediate moments after direct contact with an implanted surface.
Aspartoacylase Deficiency in the White Matter of Human Immunodeficiency Virus Encephalitis: Novel Mechanism in Axonal Damage
Authors: Surendran Sankar , Rajasankar Srinivasagam
Published: September 2011

Aspartoacylase/aminoacylase II (ASPA/ACY II) is mainly synthesized in oligodendrocytes to contribute in myelin synthesis. Although axonal damage is seen in the brain with human immunodeficiency virus encephalitis (HIVE), ASPA contribution in the pathology is not known. Immunostaining study showed that ASPA protein is reduced in the white matter of patients with HIVE compared to the control. Western blot study further confirmed ASPA deficiency in the HIVE brain compared to the control. This paper suggests that HIVE condition affects ASPA to contribute in myelin loss/axonal damage seen in the disease.
The productivity effect of informal employees: The Tshwane tyre-fitting industry as a case study
Authors: Van Zyl Gerhardus
Published: September 2011

Orientation: The article investigated the application of a suitable estimation model to measure the productivity spill over effects of informal employees.Research purpose: The aim of the article was to estimate the sign and magnitude of productivity spill over effects of informal employees when employee and firm characteristics as well as external factors were considered.Motivation for the study: The researcher thought that empirical research was appropriate because of the belief that, because of low productivity levels in the South African workplace, firms are resorting to greater numbers of informal employees in order to generate positive productivity spill over effects that would improve their levels of competitiveness.Research design, approach and methodology: The researcher constructed real data series on the applicable variables from weekly data that he collected from firms in the sample group over the sample period. He then applied the Nelen, de Grip and Fourage estimation model to determine the sign and magnitude of the spill over effects for the different employee segments.Main findings: The informal employee segment created greater positive productivity spill over effects. This was especially true for older informal employees with longer tenures and lower levels of absenteeism. External factors, like variations in market demand, improved the positive spill over effects that informal employees generated.Practical/managerial implications: Firms might reconsider the composition of their workforce if the informal employee segment consistently generated higher positive productivity spill over effects.Contribution/value-add: The researcher has introduced an estimation procedure that firms can apply to measure the productivity spill over effects of formal and informal employee segments at firm and industry level.
Crenarchaea and phytoplankton coupling in sedimentary archives: Common trigger or metabolic dependence?
Authors: Fietz Susanne , Martínez-Garcia Alfredo , Rueda Gemma , Peck Vicky L. , Huguet Carme , Escala Marina , Rosell-Melé Antoni
Published: September 2011

The concentrations of chlorins (chlorophyll transformation products indicative of phytoplankton production) and crenarchaeol (a marker for Crenarchaea abundance) are significantly positively correlated (Spearman's rank correlation coefficient rs > 0.75) in four core records from freshwater (Lake Baikal) and marine settings (Southern, Atlantic, and Arctic Oceans). This suggests a close relationship between Crenarchaea abundance and phytoplankton production. Degradation and transport mechanisms, as well as a common environmental trigger, may in part account for our observations, but these mechanisms alone cannot fully explain them. Instead our findings point to a metabolic dependence of Crenarchaea on resources released by phytoplankton, such as organic carbon or ammonium.
Depth attenuation of organic matter export associated with jelly falls
Authors: Lebrato Mario , Pahlow Markus , Oschlies Andreas , Pitt Kylie A. , Jones Daniel O. B. , Molinero Juan Carlos , Condon Robert H.
Published: September 2011

We explore the attenuation in the export ratio of jelly‐POM (particulate organic matter) with depth as a function of the decay rate, temperature, and sedimentation rate. Using data from the Vertical Transport In the Global Ocean project, we compare ratios computed with the Martin‐curve, with a particle‐based parameterization, and with sediment‐trap data. Owing to the temperature dependence of the decay rate (Q10 = 4.28), the jelly‐POM export ratio below 500 m is 20–45% larger in subpolar and temperate areas than in the tropics. Vertical migration of gelatinous zooplankton leads to a variable starting depth of a jelly fall (death depth), which governs the start of remineralization, and the fate of the biomass. Owing to the absence of observations, we employ a sinking speed matrix ranging from 100 m d‐1 to 1500 m d‐1 to represent slow‐ and fast‐sinking carcasses. The assumption of a constant decay rate k independent of temperature in other particle‐based models may not be appropriate. These results provide information for including jelly‐POM in global biogeochemical model formulations.
Wind turbines in icing conditions: performance and prediction
Authors: Dierer S. , Oechslin R. , Cattin R.
Published: September 2011

Abstract. Icing on structures is an important issue for wind energy developments in many regions of the world. Unfortunately, information about icing conditions is mostly rare due to a lack of measurements. Additionally, there is not much known about the operation of wind turbines in icing conditions. It is the aim of the current study to investigate the effect of icing on power production and to evaluate the potential of icing forecasts to help optimizing wind turbine operation. A test site with two Enercon E-82 turbines was set up in the Jura region in Switzerland in order to study the turbines' behaviour in icing conditions. Icing forecasts were performed by using an accretion model driven by results of the mesoscale weather forecast model WRF. The icing frequency at the test site is determined from pictures of a camera looking at the measurement sensors on the nacelle. The results show that the site is affected by frequent icing: 11.5 days/year of meteorological icing and 41.5 days/year of instrumental icing were observed corresponding to a factor of about four. The comparison of power production with and without blade heating shows that blade heating results in a 3.5% loss and operation without blade heating results in a 10% loss of the annual power production due to icing. Icing forecasts are performed for winter 2009/2010. Simulated and observed icing events agree well and also coincide with periods of power drop. Thus, the results suggest that icing forecasts can help to optimize the operation of wind parks in icing conditions.
Links Patrocinados como Ferramenta para Captação de Clientes na Hotelaria
Authors: Mondo Tiago Savi , Pereira Daniel Augustin , Costa Jane Iara Pereira da
Published: September 2011

Os links patrocinados desempenham a funo de aumentar a velocidade e a qualidade das respostas de buscas realizadas em sites de pesquisa na internet. Nesse contexto, os meios de hospedagem, se enquadram como integrantes do processo, j que muitas das buscas de informaes sobre destinos e empreendimentos tursticos so feitas por meio da internet. Assim, o objetivo central desse artigo identificar a ocorrncia de links patrocinados de meios de hospedagem de Santa Catarina nos buscadores Google, Terra e Uol. Esta pesquisa caracteriza-se por ser exploratria,descritiva, de carter quantitativo e do tipo documental. A amostra foi composta por 2713 links patrocinados, provenientes de buscas realizadas com os termos hotis e pousadas e ainda os termos geogrficos Florianpolis, Joinville, Blumenau, Bombinhas e Balnerio Cambori. As buscas foram realizadas do dia 10 ao dia 15 de Maro de 2010, resultando em 450 buscas. Os principais resultados demonstram que os meios de hospedagem poderiam utilizar melhor a ferramenta e que boa parte da sua utilizao de links patrocinados feita por classificados, conglomerados de sites hoteleiros e agncias de viagem online. Alm disso, levantou-se que a ferramenta mais utilizada nos fins de semana e durante os perodos vespertino e noturno. Conclui-se que a ferramenta indispensvel atualmente para o segmento turstico, servindo como auxilio distribuio dos servios do hotel e, consequentemente, ao aumento da ocupao hoteleira. DOI: 10.5585/remark.v10i1.2185
The Fentanyl/Etomidate‐Anesthetized Beagle (FEAB) Model in Safety Pharmacology Assessment
Authors: van der Linde Henk J. , Deuren Bruno Van , Somers Yves , Teisman Ard , Gallacher David J.
Published: September 2011

This unit describes a procedure for performing safety studies in the anesthetized beagle dog. Detailed are the anesthetic regime, the surgical procedure, and all materials needed to perform cardiovascular, central nervous system, and respiratory safety studies in these animals. An overview of all parameters that can be measured and calculated is provided, as are experimental protocols. Endpoints discussed include hemodynamic, electrocardiological, respiratory, arterial blood, and electroencephalogical parameters. Also presented are a formula to correct QT interval for changes in core body temperature and an overview of changes in ECG, MAP, and EEG traces that may occur during safety studies. The information provided yields a multiparametric model for performing reliable safety studies in anesthetized dogs. Curr. Protoc. Pharmacol. 54:10.13.1‐10.13.21. © 2011 by John Wiley & Sons, Inc.
Biochemical Evaluation of HCV NS3 Protease Inhibitors
Authors: Schultz Brian , Yang Huiling , Delaney William E.
Published: September 2011

This unit describes assays for characterizing the potency and mechanism of action of NS3 protease inhibitors. Determination of IC50values is described using in vitro expressed and purified NS3 protease. This assay can also be used for the rapid exploration of structure‐activity relationships. Another protocol describes using the full‐length NS3/4A complexes expressed in HCV replicon cell lines for a rapid alternative method for assessing protease activity without requiring conventional protein expression and purification. A method is then provided for determination of inhibitorKi, which more accurately assesses the potency of inhibitors compared to the IC50assay, particularly for potent inhibitors that reach the sensitivity limit for the basic IC50assay. The final protocol describes how to determine the reversibility of inhibitor binding to the enzyme, an important parameter that can affect the pharmacodynamic properties of a compound.Curr. Protoc. Pharmacol. 54:13B.7.1‐13B.7.22. © 2011 by John Wiley & Sons, Inc.
Orthotopic Models of Esophageal Carcinoma and Their Use in Drug Discovery
Authors: Gros Stephanie J.
Published: September 2011

The protocol detailed in this unit is for the establishment of an orthotopic model of human esophageal adenocarcinoma in NMRI/nu mice. The resultant tumor has high metastatic potential, spreading readily to liver, lungs, and lymph nodes. This model is useful for studying primary esophageal carcinoma, tumor biology, pathogenesis, tumor progression, metastatic homing, and the efficacy of therapeutic approaches for treating this condition. The practical use of this preclinical model for drug discovery is illustrated with data from a study on the chemotherapeutic effects of HER2‐targeted therapy. Curr. Protoc. Pharmacol. 54:14.20.1‐14.20.15. © 2011 by John Wiley & Sons, Inc.
Pre‐Clinical Mouse Models of Primary and Metastatic Pleural Cancers of the Lung and Breast and the Use of Bioluminescent Imaging to Monitor Pleural Tumor Burden
Authors: Servais Elliot L. , Colovos Christos , Kachala Stefan S. , Adusumilli Prasad S.
Published: September 2011

Malignant pleural disease (MPD) results in an estimated 150,000 cases of malignant pleural effusions (MPE) annually. The most common malignancies associated with MPD are primary malignant pleural mesothelioma (MPM) and metastatic lung cancer, breast cancer, and lymphoma. MPM is a rare, regionally aggressive malignancy whose incidence is increasing secondarily to the latency of disease progression. MPD is characteristic of advanced‐stage pleural disease and portends a grave clinical prognosis with a median survival between 3 and 12 months. Preclinical investigations conducted in flank and intraperitoneal tumor models do not fully recapitulate the pleural tumor microenvironment, and the results are not directly translatable to the clinical setting. The protocol described herein provides a mouse model of MPM and MPD from nonhematogenous tumors, resulting in reproducible tumor location, tumor progression, animal survival, and histopathology. Pleural tumor growth in this model resembles the regionally aggressive clinical course and tumor microenvironment of human pleural cancers and provides an optimal animal model to investigate MPD biology and therapies. Curr. Protoc. Pharmacol. 54:14.21.1‐14.21.18. © 2011 by John Wiley & Sons, Inc.
The Colorimetric Detection and Quantitation of Total Protein
Authors: Krohn Randall I.
Published: September 2011

Protein quantification is an important step for handling protein samples for isolation and characterization; it is a prerequisite step before submitting proteins for chromatographic, electrophoretic, or immunochemical analysis and separation. Colorimetric methods are fast, simple, and not laborious. This unit describes a number of assays able to detect protein concentrations in the low microgram to milligram per milliliter ranges in a variety of formats. Curr. Protoc. Cell Biol. 52:A.3H.1‐A.3H.28. © 2011 by John Wiley & Sons, Inc.
Janus Kinase V617F mutation in cigarette smokers
Authors: Weinberg Ido , Borohovitz Ariel , Krichevsky Svetlana , Perlman Riki , Ben‐Yehuda Arie , Ben‐Yehuda Dina
Published: September 2011

The JAK2 V617F mutation is responsible for the constitutive activation of the erythropoietin receptor signaling pathway in most cases of polycythemia vera (PV). The mutation has also been described in healthy people. As smoking may result in secondary polycythemia, the goal of this trial was to examine the effect of smoking on the prevalence of the JAK2 mutation and its correlation to erythrocytosis. The study was case–control. Hospitalized smokers (n = 81) and nonsmokers (n = 61) were recruited. Serum was drawn for complete blood count, erythropoietin, ferritin and venous blood gases. JAK2 mutation was analyzed by highly sensitive allele‐specific Quantitative Real Time PCR. The JAK2 mutation was found in 29/81 (35.8%) of smokers in comparison to only 9/61 (14.8%) of the control group (P = 0.007). The frequency of the mutation among smokers who were positive for the JAK2 mutation had a mean of 6.78 × 10−4 ± 1.08 × 10−3 vs. 1.51 × 10−4 ± 2.04 × 10−4 among nonsmokers (P = 0.027). Both frequencies are much lower than those found in PV. There was a medium correlation between older age and mutation frequency in nonsmokers (r= 0.67, P = 0.043). Hematocrit was higher in smokers (47.8 ± 6 vs. 41.7 ± 4.7, P < 0.0001), but no correlation was found to JAK2 mutation. In a cohort of hospitalized smokers and nonsmokers, JAK2 mutation was more prevalent and found in higher frequencies among smokers than nonsmokers. We suggest that accelerated erythropoiesis renders the cells susceptible to JAK2 mutation. Am. J. Hematol., 2011. © 2011 Wiley Periodicals, Inc.
Seasonal versatility in the feeding ecology of a group of titis (Callicebus coimbrai) in the northern Brazilian Atlantic Forest
Authors: Souza‐Alves João Pedro , Fontes Isadora P. , Chagas Renata R.D. , Ferrari Stephen F.
Published: September 2011

The feeding behavior of a group of titis (Callicebus coimbrai) was monitored over an annual cycle at a site in northeastern Brazil. Behavioral data were collected in scan samples (1‐min scan at 5‐min intervals), and complementary data on fruit availability and new leaf cover were collected. Feeding time accounted for 28.9% of daily activity. Fruit was the principal item of the diet (61.2% of records) and the primary category in all months except September, when it was surpassed by leaves. Young leaves were the second most important category (20.0%). The consumption of seeds and insects was prominent in November and December. Fifty‐two plant species were exploited, and the Elaeocarpaceae, Myrtaceae, Sapotaceae, and Passifloraceae provided the vast majority (86.0%) of plant feeding records. The phenological record did not provide a good measure of fruit availability, but a strong correlation (rs=0.902,P<0.0001,n=12) was found between the consumption of leaves and the exploitation of lianas each month. Lianas accounted for 28.2% of plant feeding records, and predominated between August and December. This suggests that lianas may represent a key factor in the ability of the species to tolerate the intense habitat fragmentation found throughout its geographic range. Am. J. Primatol. 73:1199–1209, 2011. © 2011 Wiley Periodicals, Inc.
Sex ratio affects sex‐specific innovation and learning in captive ruffed lemurs (Varecia variegataandVarecia rubra)
Authors: Dean Lewis G. , Hoppitt William , Laland Kevin N. , Kendal Rachel L.
Published: September 2011

Recent years have witnessed extensive research into problem solving and innovation in primates, yet lemurs have not been subjected to the same level of attention as apes and monkeys, and the social context in which novel behavior appears has rarely been considered. We gave novel foraging puzzlebox devices to seven groups of ruffed lemurs (Varecia variegataandVarecia rubra) to examine the factors affecting rates of innovation and social learning. We found, across a range of group sex ratios, that animals of the less‐represented sex were more likely to contact and solve the puzzlebox sooner than those of the more‐represented sex. We established that while some individuals were able to solve the puzzleboxes there was no evidence of social learning. Our findings are consistent with previously reported male deference as a sexual strategy, but we conclude that the need for male deference diminishes when, within a group, males are rare. Am. J. Primatol. 73:1210–1221, 2011. © 2011 Wiley Periodicals, Inc.
Protozoan parasites in group‐living primates: testing the biological island hypothesis
Authors: Chapman Colin A. , Bowman Dwight D. , Ghai Ria R. , Gogarten Jan F. , Goldberg Tony L. , Rothman Jessica M. , Twinomugisha Dennis , Walsh Chesley
Published: September 2011

A series of articles by W.J. Freeland published in the 1970s proposed that social organization and behavioral processes were heavily influenced by parasitic infections, which led to a number of intriguing hypotheses concerning how natural selection might act on social factors because of the benefits of avoiding parasite infections. For example, Freeland [1979] showed that all individuals within a given group harbored identical gastrointestinal protozoan faunas, which led him to postulate that social groups were akin to “biological islands” and suggest how this isolation could select specific types of ranging and dispersal patterns. Here, we reexamine the biological island hypothesis by quantifying the protozoan faunas of the same primate species examined by Freeland in the same location; our results do not support this hypothesis. In contrast, we quantified two general changes in protozoan parasite community of primates in the study area of Kibale National Park, Uganda, over the nearly 35 years between sample collections: (1) the colobines found free of parasites in the early 1970s are now infected with numerous intestinal protozoan parasites and (2) groups are no longer biological islands in terms of their protozoan parasites. Whatever the ultimate explanation for these changes, our findings have implications for studies proposing selective forces shaping primate behavior and social organization. Am. J. Primatol. 74:510–517, 2012. © 2011 Wiley Periodicals, Inc.
Cyclin‐dependent kinase inhibitor p21, via its C‐terminal domain, is essential for resolution of murine inflammatory arthritis
Authors: Mavers Melissa , Cuda Carla M. , Misharin Alexander V. , Gierut Angelica K. , Agrawal Hemant , Weber Evan , Novack Deborah Veis , Haines G. Kenneth , Balomenos Dimitrios , Perlman Harris
Published: September 2011

Objective

The mechanism responsible for persistent synovial inflammation in rheumatoid arthritis (RA) is unknown. Previously, we demonstrated that expression of the cyclin‐dependent kinase inhibitor p21 is reduced in synovial tissue from RA patients compared to osteoarthritis patients and that p21 is a novel suppressor of the inflammatory response in macrophages. The present study was undertaken to investigate the role and mechanism of p21‐mediated suppression of experimental inflammatory arthritis.

Methods

Experimental arthritis was induced in wild‐type or p21−/− (C57BL/6) mice, using the K/BxN serum–transfer model. Mice were administered p21 peptide mimetics as a prophylactic for arthritis development. Lipopolysaccharide‐induced cytokine and signal transduction pathways in macrophages that were treated with p21 peptide mimetics were examined by Luminex‐based assay, flow cytometry, or enzyme‐linked immunosorbent assay.

Results

Enhanced and sustained development of experimental inflammatory arthritis, associated with markedly increased numbers of macrophages and severe articular destruction, was observed in p21−/− mice. Administration of a p21 peptide mimetic suppressed activation of macrophages and reduced the severity of experimental arthritis in p21‐intact mice only. Mechanistically, treatment with the p21 peptide mimetic led to activation of the serine/threonine kinase Akt and subsequent reduction of the activated isoform of p38 MAPK in macrophages.

Conclusion

These are the first reported data to reveal that p21 has a key role in limiting the activation response of macrophages in an inflammatory disease such as RA. Thus, targeting p21 in macrophages may be crucial for suppressing the development and persistence of RA.
Iron‐Catalyzed CH Bond Activation for the ortho‐Arylation of Aryl Pyridines and Imines with Grignard Reagents
Authors: Yoshikai Naohiko , Asako Sobi , Yamakawa Takeshi , Ilies Laurean , Nakamura Eiichi
Published: September 2011

Direct arylation of the ortho‐CH bond of an aryl pyridine or an aryl imine with an aryl Grignard reagent has been achieved by using an iron‐diamine catalyst and a dichloroalkane as an oxidant in a short reaction time (e.g., 5 min) under mild conditions (0 °C). The use of an aromatic co‐solvent, such as chlorobenzene and benzene, and slow addition of the Grignard reagent are essential for the high efficiency of the reaction. The present arylation reaction has distinct merits over the previously developed reaction that used an arylzinc reagent, such as its reaction rate and atom economy. Selective CH bond activation occurs in the presence of a leaving group, such as a tosyloxy, chloro, and bromo group. Studies on a stoichiometric reaction and kinetic isotope effects shed light on the reaction intermediate and the CH bond‐activation step.
Cell line specific control of polyethylenimine‐mediated transient transfection optimized with “Design of experiments” methodology
Authors: Thompson Ben C. , Segarra Camille R. J. , Mozley Olivia L. , Daramola Olalekan , Field Ray , Levison Peter R. , James David C.
Published: September 2011

We describe a design of experiments (DoE) response surface modeling strategy to optimize the concentration of basal variables underpinning polyethylenimine (PEI) mediated transfection of different CHO‐K1 derived parental cell populations in a chemically defined medium, specifically the relative concentration of linear 25 kD PEI, host CHO cells and plasmid DNA. Using recombinant secreted alkaline phosphatase (SEAP) reporter activity as the modeled response, a discrete simple maximum was predicted for each CHO host cell population. Differences between the modeled optima derived from host cell specific differences in PEI cytotoxicity, such that the PEI:cell interaction effectively limited PEI‐DNA polyplex load at a relatively constant PEI:DNA ratio. However, across the three CHO host cell populations, SEAP reporter production was not proportional to plasmid DNA input at the host cell specific predicted basal variable optima. A 10‐fold variation in SEAP reporter output per mass of plasmid DNA delivered was observed. To determine the cellular basis of this difference in transient productivity, host CHO cells were transfected with fluorescently labeled polyplexes followed by flow cytometric analysis. Each CHO host cell population exhibited a distinct functional phenotype, varying in the extent of PEI‐DNA polyplex binding to the cell surface and degree of polyplex internalization. SEAP production was directly proportional to the level of polyplex internalization and heparan sulfate proteoglycan level. Taken together, these data show that choice of host CHO cell line is a critical parameter, which should rationally precede cell line specific transient production platform design using DoE methodology. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Modifying Cage Structures in Metal–Organic Polyhedral Frameworks for H2 Storage
Authors: Yan Yong , Blake Alexander J. , Lewis William , Barnett Sarah A. , Dailly Anne , Champness Neil R. , Schröder Martin
Published: September 2011

Three isostructural metal–organic polyhedral cage based frameworks (denoted NOTT‐113, NOTT‐114 and NOTT‐115) with (3,24)‐connected topology have been synthesised by combining hexacarboxylate isophthalate linkers with {Cu2(RCOO)4} paddlewheels. All three frameworks have the same cuboctahedral cage structure constructed from 24 isophthalates from the ligands and 12 {Cu2(RCOO)4} paddlewheel moieties. The frameworks differ only in the functionality of the central core of the hexacarboxylate ligands with trimethylphenyl, phenylamine and triphenylamine moieties in NOTT‐113, NOTT‐114 and NOTT‐115, respectively. Exchange of pore solvent with acetone followed by heating affords the corresponding desolvated framework materials, which show high BET surface areas of 2970, 3424 and 3394 m2 g−1 for NOTT‐113, NOTT‐114 and NOTT‐115, respectively. Desolvated NOTT‐113 and NOTT‐114 show high total H2 adsorption capacities of 6.7 and 6.8 wt %, respectively, at 77 K and 60 bar. Desolvated NOTT‐115 has a significantly higher total H2 uptake of 7.5 wt % under the same conditions. Analysis of the heats of adsorption (Qst) for H2 reveals that with a triphenylamine moiety in the cage wall, desolvated NOTT‐115 shows the highest value of Qst for these three materials, indicating that functionalisation of the cage walls with more aromatic rings can enhance the H2/framework interactions. In contrast, measurement of Qst reveals that the amine‐substituted trisalkynylbenzene core used in NOTT‐114 gives a notably lower H2/framework binding energy.
New Energetic Polynitro Cyclic Esters: Ammonium, Hydrazinium, and Hydroxylammonim Salts of Polynitramines
Authors: Singh Rajendra P. , Shreeve Jean'ne M.
Published: September 2011

Reaction of 2,2‐dinitro‐1,3‐propanediol (1) with oxalyl dichloride or malonyl dichloride in refluxing ether led to the formation of cyclic dinitro‐containing esters (2,3) in very good yields. Compounds2and3were also isolated in similar yields by the treatment of1with oxalic acid or malonic acid in trifluoroacetic anhydride at room temperature. Nitration of3with fuming nitric acid resulted in the corresponding trinitro cyclic ester4in 70 % isolated yield. Treatment of1with a large excess of methanolic ammonia gave impure 2,2‐dinitro‐1,3‐diaminopropane (5). Polynitraamines,7and11, were treated with aqueous ammonia, hydrazine monohydrate or hydroxylamine in methanol at room temperature to obtain their corresponding salts8–10and12–14, respectively, in excellent isolated yields. All new compounds were characterized by IR, NMR spectroscopy (1H,13C,15N), DSC, and elemental analysis. Their energetic properties, such as impact sensitivity, detonation velocity, and detonation pressure were also determined and compared with existing energetic compounds, such as PETN (pentaerythritol tetranitrate), RDX (1,3,5‐trinitro‐1,3,5‐triazacyclohexane), and TNT (trinitrotoluene).