bluuebunny/crossref_metadata_2025 huggingface dataset:
Engaging with Armed Groups: A Human Rights Field Perspective from Nepal
Authors: Rawski Frederick
Published: April 2010

Theoretical uncertainty surrounding whether and to what extent non-state actors are obligated to abide by international human rights and humanitarian norms has impacted the policies and practices of human rights field presences, including those of the United Nations. The paper presents the case of Nepal, where international organizations have been forced by circumstance to engage with a range of non-state armed groups, to illustrate how current limitations of human rights law regarding non-state actors impact the work of human rights field monitors. Drawing upon the global administrative law paradigm, it argues for the development of a 'soft law' framework governing human rights aspects of international organizations' engagement with armed groups.
Corporate Location, Concentration and Performance: Large Company Headquarters in the Australian Urban System
Authors: Tonts Matthew , Taylor Michael
Published: April 2010

The paper examines the geography of corporate headquarters in the Australian urban system, giving consideration to their location, control of capital and performance. The paper argues that, while considerable recent attention has been given to global cities in the networks of corporate power, the spatial organisation of company headquarters remains important within national urban systems. In the case of Australia, Sydney and Melbourne dominate the corporate landscape, although the smaller cities of Adelaide, Brisbane and Perth occupy important niches. The paper demonstrates that there are clear variations in the relative performance of companies across the Australian urban system. There is also a pattern of sectoral specialisation within Australia’s cities which, in part, helps to explain the different development trajectories and relative levels of corporate performance. The paper concludes by exploring some of the implications of the study for the understanding of urban systems.
Cosmopolitanism, Geographical Imaginaries and Belonging in North London
Authors: Devadason Ranji
Published: April 2010

Cosmopolitanism has been described as the cultural habitus of globalisation. It is therefore, albeit defined somewhat loosely, often associated with ethnically diverse, global cities. This paper considers the extent to which London engenders cosmopolitan values amongst its residents. It draws on survey data from the LOCAL MULTIDEM study of minorities’ political participation to address these themes. The analysis examines perceptions of respect, belonging and geographical imaginaries—amongst established minorities and the ethnic majority—in north London. It is argued that cosmopolitan ethics are transformative and dialectical and, critically, cannot remain the preserve of the privileged in multi-ethnic neighbourhoods. The analysis presented demonstrates that a sense of belonging and cosmopolitan imaginaries are not evenly accessed by different ethnic groups; notably, that Bangladeshi Londoners who are born and bred in the city are less likely to appropriate these discourses than Caribbean, Indian or White residents.
Soldiers, Scholars, and Spies: Combining Smarts and Secrets
Authors: Miller Bowman H.
Published: April 2010

The worlds of intelligence, the military, and academe reflect very different, at times conflicting, cultures, ethical norms, and priorities. This examination assesses the evolution of the sometimes fractious relationship between scholars and government military and intelligence officials, disputes among them, and cases of convergence and critical collaboration.To be effective in policy and decision making, the U.S. government needs the best information and thinking it can obtain. Much of that expertise resides in academia. For their part, social scientists and members of the healing professions, in particular, must be attentive to the moral, intellectual, and ethical dictates of their professions when participating in support of government employment of coercive military force and extraordinary means of intelligence acquisition. Finding the appropriate balance between helping safeguard national security and upholding the tenets of professional integrity is a joint task of soldiers, scholars, and spies.
Gender Disparity and HIV Risk Among Young Black Women in College: A Literature Review
Authors: Alleyne Binta , Gaston Gina
Published: April 2010

This review article explores the issue of the gender-ratio imbalance among African American college students and its effect on the HIV risk behavior of young Black women in college. The theory of gender and power is used as a conceptual framework. The authors strive to show that there is a significant need for practitioners and researchers to develop and provide holistic gender-specific and culturally competent HIV prevention services for this unique population that reduce power differentials and challenge social norms. The terms African American and Black are used interchangeably to be inclusive of all individuals of African descent.
Graduating Social Work Students’ Perspectives on Domestic Violence
Authors: Black Beverly M. , Weisz Arlene N. , Bennett Larry W.
Published: April 2010

This article reports the findings of a qualitative study that examined 124 social work students’ views on the causes and dynamics of domestic violence and their recommended interventions in a case scenario. Most students graduated from the master of social work (MSW) program with a mental health perspective on domestic violence. Only a small percentage were aware of specific interventions for domestic violence and many continued to attribute domestic violence to mental health and substance abuse problems in the victim and perpetrator. Graduates with domestic violence experience varied little from their peers in suggested domestic violence interventions or comments about the causes of domestic violence. The findings suggest that a deliberate, focused attempt to inform students about domestic violence is needed, rather than a reliance on general MSW courses.

This article presents research on social work graduates’ views on the causes of domestic violence and interventions in domestic violence cases. Domestic violence continues to be a multifaceted problem that an estimated 22% of women experience over their lifetimes (Thoennes & Tjaden, 2000). Many people believe that a woman’s behavior can contribute to intimate partner violence, some believe that women want to be abused, and a large majority believe that women can easily leave abusive relationships (Worden & Carlson, 2005). Overall, most people believe that the causes of domestic violence are rooted in the individual and family; few people view domestic violence as a problem with roots in society or culture (Worden & Carlson, 2005).
Polyps, Plankton, and Passages: Mythopoeic Islands and Long-Memoried Seas
Authors: Tynan Maeve
Published: April 2010

Derek Walcott’s poetics balance on the generative lines of tension drawn between the desire to journey away from his island home and the yearning for homecoming. The poet is in many sense of the word a traveler, both physically in his journeys across the globe in search of employment and greater publishing opportunities, and mentally in the much-remarked upon allusiveness of his writing. It is unsurprising therefore, that the trope of the sea operates as a polyvalent figure within his poetic idiom. He reads the sea as a repository of the past, the swash and backwash of recurrent tides bearing witness to the continual inscription and erasure of time and thus possessive of atemporality; it is a palimpsestic record in which successive ages are contained. A blank canvas on which everything or nothing can be read, the sea for Walcott is a shifting narrative that resists the fixing impetus of Atlantic historiography, a concept reaching fullest realization in the poem The Sea Is History. For the writer living on a small island this continual sea presence can promote a feeling of provincialism, of being “castaway” on “islands [that] have drifted from anchorage,” far from the metropolitan centers of publishing houses and readerships. The outward gaze can, conversely instill an anxiety of perceived betrayal of the island home. Walcott’s identification of a “mythopoeic coast” in the poem Origins calls upon the aid of mythological analogy and imagination to fill in the gaps of memory caused by the trauma of the Middle Passage crossing. In the figure of the polyp, the postcolonial poet finds the metaphor for local, self-authorizing creation that conjoins the twin desire for routes and roots.
Shattering Silence: Exploring Barriers to Disclosure for African American Sexual Assault Survivors
Authors: Tillman Shaquita , Bryant-Davis Thema , Smith Kimberly , Marks Alison
Published: April 2010

National-, community-, and college-based studies have documented the high prevalence of sexual assault among African American women. Although African American women experience sexual assault at alarming rates, they are less likely to disclose or seek help in the aftermath of sexual assault. The purpose of this literature review is to provide a critique of the current literature examining the barriers to disclosure for African American women, such as intrapsychic factors, the damaging effect of an unsupportive response to initial disclosure, stigmatization of African American female sexuality, apprehension regarding racism, and racial loyalty. The authors provide a summary of the literature, gaps in current empirical studies, and needs for future study. Culturally relevant intervention recommendations are described. Finally, implications for sexual assault policy are provided.
Clinical outcome of carpal tunnel release with and without opposition transfer
Authors: Uemura T. , Hidaka N. , Nakamura H.
Published: April 2010

We studied the clinical outcome in 24 patients (25 hands) with carpal tunnel syndrome characterized by severe thenar muscle atrophy, who were treated by open carpal tunnel release alone (10 hands) or with simultaneous opposition transfer (15 hands). We also compared the surgical results in patients younger or older than 65 years. Pain and numbness improved after open carpal tunnel release but more so in younger patients. Activities of daily living also improved with and without opposition transfer; however, certain activities such as fastening hooks or buttons, using chopsticks and turning a key in a lock improved more with opposition transfer. Grip strength did not improve, with or without opposition transfer, regardless of the patients’ ages. Pinch strength improved more with opposition transfer.
Proposal of welding consumable of triplex stainless steel for ultrahigh strength steel
Authors: Saida K. , Nishimoto K. , Ogawa K. , Okaguchi S. , Fujiwara K.
Published: April 2010

In order to develop a new welding consumable for the ultrahigh strength HT980 steel, the microstructural characteristics, tensile properties, low temperature toughness and weld cracking susceptibility were evaluated for triplex stainless steel weld metals. Steels of 13Cr–0·5Mo–Ni with varying contents of Ni and C were investigated. The amount of retained austenite increased with increasing Ni and C contents. The tensile strengths of the weld metals decreased while their impact absorbed energies increased with increasing amount of retained austenite. Hot cracking susceptibility of the weld metals minimised in that metal containing 40–50% δ-ferrite at the completion of solidification. The triplex stainless steel weld metals, except for those containing lower amounts of retained austenite, were negligibly susceptible to cold cracking. It may be possible to design a preheat free welding consumable from a triplex stainless steel in the compositional range containing 20–40% of retained austenite.
Study on ac-PMIG welding of AZ31B magnesium alloy
Authors: Song G. , Wang P. , Liu L. M.
Published: April 2010

Welding of AZ31B magnesium alloy is carried out using alternating current pulsed metal inert gas (ac-PMIG) welding with 1·6 mm diameter of filler wire. Typical current waveform is used to make sure arc given an accurate energy input into filler wire. The arc characteristics, metal transfer forms, microstructure and mechanical property of ac-PMIG welding of AZ31B magnesium are investigated. The results show that a stable welding procedure and continuous joints can be obtained easily under a wide range of welding parameters. The most important factors for ac-PMIG welding are negative electrode (EN) ratio and pulse rework current, which give an accurate energy input into filler wire. The grain in fusion zone is much finer and more uniform, and grain size does not grow significantly in the heat affected zone compared with base metal. The average ultimate tensile strength of weld beads is 97·2% of base metal.
Constants for hot deformation constitutive models for recent experimental data
Authors: Tello K. E. , Gerlich A. P. , Mendez P. F.
Published: April 2010

This paper presents previously unavailable constants for the Sellars and Tegart constitutive model for hot metalworking. The materials considered are aluminium alloys 2024, 5083, 6061, 7050, 7075 and 356, carbon steel 1018, stainless steel 304, titanium alloy 6Al–4V, and magnesium alloys AZ31 and AZ61. These materials and their mechanical properties at high temperature are of great interest for latest generation manufacturing processes involving deformation to accomplish solid state joining, such as friction stir welding, cold spray and magnetic impulse welding. The results are also useful to model established processes, such as hot rolling, forging and creep. The methodology used to obtain the constants consists on non-linear regressions based on partial data sets as it was conducted previously. The input data were obtained from published values for hot compression experiments. All regressions presented here have a coefficient of determination R2>0·95. When possible, the results obtained were compared to previous published regressions.
Static and fatigue performance of weld bonded dual phase steel sheets
Authors: Sam S. , Shome M.
Published: April 2010

Lap joints of dual phase steel sheets of 1·0 mm were prepared by adhesive bonding, spot welding and weld bonding processes using a one component epoxy base structural adhesive. Mechanical properties of the joints were evaluated by tensile shear and fatigue tests. The size of the weld nugget for both spot weld and weld bond was measured for different welding parameters (current, time) and compared. For identical welding parameters, weld bonded nuggets exhibit higher nugget diameter. Tensile shear strength of weld bonded joints is 40 and 58% higher than spot welded joints and 15 and 39% higher than adhesive bonded joints and for DP590 and DP780 steels respectively. Considering 106 cycles, the endurance limit of weld bonded joint is much higher than spot welded joint but smaller than adhesive bonded joints. Overall the performance of weld bonded joints is superior to those of resistance spot welding.
Phase 2 and pharmacodynamic study of oral forodesine in patients with advanced, fludarabine-treated chronic lymphocytic leukemia
Authors: Balakrishnan Kumudha , Verma Dushyant , O'Brien Susan , Kilpatrick John Michael , Chen Yuling , Tyler Brenita F. , Bickel Susan , Bantia Shanta , Keating Michael J. , Kantarjian Hagop , Gandhi Varsha , Ravandi Farhad
Published: April 2010

Forodesine is a new and potent purine nucleoside phosphorylase (PNP) inhibitor. Patients with chronic lymphocytic leukemia (CLL) with primary resistance to fludarabine-based therapy or with progressive disease were eligible for oral forodesine (200 mg/d) for up to 24 weeks. Eight patients with median lymphocyte count of 35.9 × 109/L and median serum β2 microglobulin level of 6.45 mg/L were treated. Six had Rai stage III to IV and were previously heavily treated (median prior therapy = 5). Two had transient decrease in lymphocyte count to normal, whereas in 5, disease progressed. Adverse events were mild. Steady-state level of forodesine ranged from 200 to 1300nM and did not reach desired 2μM level. PNP inhibition ranged from 57% to 89% and steady-state 2′-deoxyguanosine (dGuo) concentration median was 1.8μM. Intracellular deoxyguanosine triphosphate (dGTP) increase was very modest, from median of 6μM to 10μM. Compared with in vivo, in vitro incubations of CLL lymphocytes with 10 or 20μM dGuo and forodesine (2μM) resulted in accumulation of higher levels of dGTP (40-250μM) which resulted in increase in apoptosis. Forodesine has biologic activity in CLL; pharmacodynamic parameters suggest that an alternate dosing schedule and/or higher doses to achieve greater intracellular dGTP may be beneficial in this patient population. This study is registered at www.clinicaltrials.gov as #NCT00289549.
TassDB2 - A comprehensive database of subtle alternative splicing events
Authors: Sinha Rileen , Lenser Thorsten , Jahn Niels , Gausmann Ulrike , Friedel Swetlana , Szafranski Karol , Huse Klaus , Rosenstiel Philip , Hampe Jochen , Schuster Stefan , Hiller Michael , Backofen Rolf , Platzer Matthias
Published: April 2010

Background

Subtle alternative splicing events involving tandem splice sites separated by a short (2-12 nucleotides) distance are frequent and evolutionarily widespread in eukaryotes, and a major contributor to the complexity of transcriptomes and proteomes. However, these events have been either omitted altogether in databases on alternative splicing, or only the cases of experimentally confirmed alternative splicing have been reported. Thus, a database which covers all confirmed cases of subtle alternative splicing as well as the numerous putative tandem splice sites (which might be confirmed once more transcript data becomes available), and allows to search for tandem splice sites with specific features and download the results, is a valuable resource for targeted experimental studies and large-scale bioinformatics analyses of tandem splice sites. Towards this goal we recently set up TassDB (Tandem Splice Site DataBase, version 1), which stores data about alternative splicing events at tandem splice sites separated by 3 nt in eight species.

Description

We have substantially revised and extended TassDB. The currently available version 2 contains extensive information about tandem splice sites separated by 2-12 nt for the human and mouse transcriptomes including data on the conservation of the tandem motifs in five vertebrates. TassDB2 offers a user-friendly interface to search for specific genes or for genes containing tandem splice sites with specific features as well as the possibility to download result datasets. For example, users can search for cases of alternative splicing where the proportion of EST/mRNA evidence supporting the minor isoform exceeds a specific threshold, or where the difference in splice site scores is specified by the user. The predicted impact of each event on the protein is also reported, along with information about being a putative target for the nonsense-mediated decay (NMD) pathway. Links are provided to the UCSC genome browser and other external resources.

Conclusion

TassDB2, available via http://www.tassdb.info, provides comprehensive resources for researchers interested in both targeted experimental studies and large-scale bioinformatics analyses of short distance tandem splice sites.
Adaptive coarse-grained Monte Carlo simulation of reaction and diffusion dynamics in heterogeneous plasma membranes
Authors: Collins Stuart , Stamatakis Michail , Vlachos Dionisios G
Published: April 2010

Background

An adaptive coarse-grained (kinetic) Monte Carlo (ACGMC) simulation framework is applied to reaction and diffusion dynamics in inhomogeneous domains. The presented model is relevant to the diffusion and dimerization dynamics of epidermal growth factor receptor (EGFR) in the presence of plasma membrane heterogeneity and specifically receptor clustering. We perform simulations representing EGFR cluster dissipation in heterogeneous plasma membranes consisting of higher density clusters of receptors surrounded by low population areas using the ACGMC method. We further investigate the effect of key parameters on the cluster lifetime.

Results

Coarse-graining of dimerization, rather than of diffusion, may lead to computational error. It is shown that the ACGMC method is an effective technique to minimize error in diffusion-reaction processes and is superior to the microscopic kinetic Monte Carlo simulation in terms of computational cost while retaining accuracy. The low computational cost enables sensitivity analysis calculations. Sensitivity analysis indicates that it may be possible to retain clusters of receptors over the time scale of minutes under suitable conditions and the cluster lifetime may depend on both receptor density and cluster size.

Conclusions

The ACGMC method is an ideal platform to resolve large length and time scales in heterogeneous biological systems well beyond the plasma membrane and the EGFR system studied here. Our results demonstrate that cluster size must be considered in conjunction with receptor density, as they synergistically affect EGFR cluster lifetime. Further, the cluster lifetime being of the order of several seconds suggests that any mechanisms responsible for EGFR aggregation must operate on shorter timescales (at most a fraction of a second), to overcome dissipation and produce stable clusters observed experimentally.
WaveletQuant, an improved quantification software based on wavelet signal threshold de-noising for labeled quantitative proteomic analysis
Authors: Mo Fan , Mo Qun , Chen Yuanyuan , Goodlett David R , Hood Leroy , Omenn Gilbert S , Li Song , Lin Biaoyang
Published: April 2010

Background

Quantitative proteomics technologies have been developed to comprehensively identify and quantify proteins in two or more complex samples. Quantitative proteomics based on differential stable isotope labeling is one of the proteomics quantification technologies. Mass spectrometric data generated for peptide quantification are often noisy, and peak detection and definition require various smoothing filters to remove noise in order to achieve accurate peptide quantification. Many traditional smoothing filters, such as the moving average filter, Savitzky-Golay filter and Gaussian filter, have been used to reduce noise in MS peaks. However, limitations of these filtering approaches often result in inaccurate peptide quantification. Here we present the WaveletQuant program, based on wavelet theory, for better or alternative MS-based proteomic quantification.

Results

We developed a novel discrete wavelet transform (DWT) and a 'Spatial Adaptive Algorithm' to remove noise and to identify true peaks. We programmed and compiled WaveletQuant using Visual C++ 2005 Express Edition. We then incorporated the WaveletQuant program in the Trans-Proteomic Pipeline (TPP), a commonly used open source proteomics analysis pipeline.

Conclusions

We showed that WaveletQuant was able to quantify more proteins and to quantify them more accurately than the ASAPRatio, a program that performs quantification in the TPP pipeline, first using known mixed ratios of yeast extracts and then using a data set from ovarian cancer cell lysates. The program and its documentation can be downloaded from our website at http://systemsbiozju.org/data/WaveletQuant.
Divergence of imprinted genes during mammalian evolution
Authors: Hutter Barbara , Bieg Matthias , Helms Volkhard , Paulsen Martina
Published: April 2010

Background

In contrast to the majority of mammalian genes, imprinted genes are monoallelically expressed with the choice of the active allele depending on its parental origin. Due to their special inheritance patterns, maternally and paternally expressed genes might be under different evolutionary pressure. Here, we aimed at assessing the evolutionary history of imprinted genes.

Results

In this study, we investigated the conservation of imprinted genes in vertebrate genomes and their exposition to natural selection. In a genome-wide comparison, orthologs of imprinted genes show a stronger divergence on cDNA and protein level in mammals. This pattern is most pronounced for maternally expressed genes in rodents in comparison to their non-rodent orthologs. The divergence is not attributable to increased mutation of CpG positions. It is contrasted by strong conservation of paternally expressed genes in mouse and rat. Interestingly, we found that the early divergence of imprinted genes was accompanied by an unusually strict conservation of their paralogs.

Conclusions

The apparent degeneration of maternally expressed genes may reflect a relaxation of selective pressure due to counteracting effects on maternal and embryonic fitness. Functional redundancy provided by the presence of highly conserved (non-imprinted) paralogs may have facilitated the divergence. Moreover, intensification of imprinting in modern rodents seems to have shifted the evolutionary fate of imprinted genes towards strong purifying selection.
Syphilis epidemiology in Norway, 1992-2008: resurgence among men who have sex with men
Authors: Jakopanec Irena , Grjibovski Andrej M , Nilsen Øivind , Aavitsland Preben
Published: April 2010

Background

In recent years, the number of syphilis cases has stabilised in many countries of Western Europe, however several countries have reported increases among men who have sex with men (MSM). The aim of this article was to describe the epidemiology of early syphilis in Norway in 1992-2008.

Methods

Cases of early syphilis and congenital syphilis reported to the Norwegian Surveillance System for Communicable Diseases (MSIS) 1992-2008 were described by route of transmission, gender, age, birthplace, stage of disease, HIV co-infection, source partner and place of infection.

Results

The incidence of reported syphilis ranged from 0.05 (1992) to 1.50 (2002) per 100 000 person-years. Of 562 cases reported to MSIS during the study period, 62% were men infected by another man. The proportion of those, infected homosexually increased from 0 (1992-1994) to 77% (2008). Most of them were Norwegians (83%). The proportion of HIV co-infection among homosexually infected increased over time and reached 39% in 2008. The majority reported being infected by a casual partner (73%) and in the municipality of Oslo (72%). Of 152 heterosexually infected men 64% were Norwegians; 51% were infected by casual contacts and 20% by commercial sex workers; 73% were infected abroad. Among 56 women, 57% were Norwegians, 57% were infected by a steady partner and 40% were infected abroad. Almost half (46%) were diagnosed in the early latent stage. Four cases had congenital syphilis, two of whom were adopted from abroad.

Conclusions

Syphilis is rare in Norway, but MSM represent almost two thirds of cases. The increase of HIV co-infected cases among MSM may enhance transmission of both infections. We recommend sexually active MSM to be tested for syphilis 2-4 times a year. Due to its variable clinical course, syphilis might be difficult to recognise at an early stage among women in a low-prevalence population. We estimate current practice of prenatal screening in Norway as sufficient.
Low aerobic mitochondrial energy metabolism in poorly- or undifferentiated neuroblastoma
Authors: Feichtinger Rene' G , Zimmermann Franz , Mayr Johannes A , Neureiter Daniel , Hauser-Kronberger Cornelia , Schilling Freimut H , Jones Neil , Sperl Wolfgang , Kofler Barbara
Published: April 2010

Background

Succinate dehydrogenase (SDH) has been associated with carcinogenesis in pheochromocytoma and paraganglioma. In the present study we investigated components of the oxidative phosphorylation system in human neuroblastoma tissue samples.

Methods

Spectrophotometric measurements, immunohistochemical analysis and Western blot analysis were used to characterize the aerobic mitochondrial energy metabolism in neuroblastomas (NB).

Results

Compared to mitochondrial citrate synthase, SDH activity was severely reduced in NB (n = 14) versus kidney tissue. However no pathogenic mutations could be identified in any of the four subunits of SDH. Furthermore, no genetic alterations could be identified in the two novel SDH assembly factors SDHAF1 and SDH5. Alterations in genes encoding nfs-1, frataxin and isd-11 that could lead to a diminished SDH activity have not been detected in NB.

Conclusion

Because downregulation of other complexes of the oxidative phosphorylation system was also observed, a more generalized reduction of mitochondrial respiration seems to be present in neuroblastoma in contrast to the single enzyme defect found in hereditary pheochromocytomas.
Association between TGFBR1*6A and osteosarcoma: A Chinese case-control study
Authors: Hu Yun-Sheng , Pan Yong , Li Wen-Hai , Zhang Yong , Li Jun , Ma Bao-An
Published: April 2010

Background

TGFBR1*6A is a common hypomorphic variant of transforming growth factor β receptor 1 (TGFBR1). TGFBR1*6A is associated with an increased cancer risk, but the association of this polymorphism with osteosarcoma remains unknown. We have measured the frequency of TGFBR1*6A variants in osteosarcoma cases and controls.

Methods

Our case-control study is based on 168 osteosarcoma patients and 168 age- and gender-matched controls. Blood samples were obtained and the TGFBR1*6A variant determined by PCR amplification and DNA sequencing. The odds ratio (OR) and 95% confidence interval (95% CI) for the TGFBR1*6A polymorphism were calculated by unconditional logistic regression, adjusted for both age and gender. Three models - dominant, additive and recessive - were used to analyze the contribution of the TGFBR1*6A variant to osteosarcoma susceptibility.

Results

Heterozygotic and homozygotic TGFBR1*6A variants represented 50.4% and 6.0% of the 168 cases, whereas the controls had 18. 5% and 1.3%, respectively. ORs for homozygosity and heterozygosity of the TGFBR1*6A allele were 4.6 [95% CI, 2.33-7.97] and 2.9 [95% CI, 1.59-5.34] in the additive model. There were significant increases in the TGFBR1*6A variants in osteosarcoma cases compared to control in all 3 models. Further analysis showed that TGFBR1*6A genotypes were not associated with gender, age, or tumor location. However, TGFBR1*6A was significantly associated with less metastasis.

Conclusions

TGFBR1*6A, a dominant polymorphism of TGFBR1, is associated with increased susceptibility and metastasis spread of osteosarcoma.
Enhanced fluorescent properties of an OmpT site deleted mutant of Green Fluorescent Protein
Authors: Salunkhe Shardul S , Raiker Veena A , Rewanwar Sachin , Kotwal Prakash , Kumar Avijeet , Padmanabhan Sriram
Published: April 2010

Background

The green fluorescent protein has revolutionized many areas of cell biology and biotechnology since it is widely used in determining gene expression and for localization of protein expression. Expression of recombinant GFP in E. coli K12 host from pBAD24M-GFP construct upon arabinose induction was significantly lower than that seen in E. coli B cells with higher expression at 30°C as compared to 37°C in E. coli K12 hosts. Since OmpT levels are higher at 37°C than at 30°C, it prompted us to modify the OmpT proteolytic sites of GFP and examine such an effect on GFP expression and fluorescence. Upon modification of one of the two putative OmpT cleavage sites of GFP, we observed several folds enhanced fluorescence of GFP as compared to unmodified GFPuv (Wild Type-WT). The western blot studies of the WT and the SDM II GFP mutant using anti-GFP antibody showed prominent degradation of GFP with negligible degradation in case of SDM II GFP mutant while no such degradation of GFP was seen for both the clones when expressed in BL21 cells. The SDM II GFP mutant also showed enhanced GFP fluorescence in other E. coli K12 OmpT hosts like E. coli JM109 and LE 392 in comparison to WT GFPuv. Inclusion of an OmpT inhibitor, like zinc with WT GFP lysate expressed from an E. coli K12 host was found to reduce degradation of GFP fluorescence by two fold.

Results

We describe the construction of two GFP variants with modified putative OmpT proteolytic sites by site directed mutagenesis (SDM). Such modified genes upon arabinose induction exhibited varied degrees of GFP fluorescence. While the mutation of K79G/R80A (SDM I) resulted in dramatic loss of fluorescence activity, the modification of K214A/R215A (SDM II) resulted in four fold enhanced fluorescence of GFP.

Conclusions

This is the first report on effect of OmpT protease site modification on GFP fluorescence. The wild type and the GFP variants showed similar growth profile in bioreactor studies with similar amounts of recombinant GFP expressed in the soluble fraction of the cell. Our observations on higher levels of fluorescence of SDM II GFP mutant over native GFPuv in an OmpT+ host like DH5α, JM109 and LE392 at 37°C reiterates the role played by host OmpT in determining differences in fluorescent property of the expressed GFP. Both the WT GFP and the SDM II GFP plasmids in E. coli BL21 cells showed similar expression levels and similar GFP fluorescent activity at 37°C. This result substantiates our hypothesis that OmpT protease could be a possible factor responsible for reducing the expression of GFP at 37°C for WT GFP clone in K12 hosts like DH5α, JM109, LE 392 since the levels of GFP expression of SDM II clone in such cells at 37°C is higher than that seen with WT GFP clone at the same temperature.
Differential alterations of the concentrations of endocannabinoids and related lipids in the subcutaneous adipose tissue of obese diabetic patients
Authors: Annuzzi Giovanni , Piscitelli Fabiana , Di Marino Lucrezia , Patti Lidia , Giacco Rosalba , Costabile Giuseppina , Bozzetto Lutgarda , Riccardi Gabriele , Verde Roberta , Petrosino Stefania , Rivellese Angela A , Di Marzo Vincenzo
Published: April 2010

Background

The endocannabinoids, anandamide and 2-AG, are produced by adipocytes, where they stimulate lipogenesis via cannabinoid CB1 receptors and are under the negative control of leptin and insulin. Endocannabinoid levels are elevated in the blood of obese individuals and nonobese type 2 diabetes patients. To date, no study has evaluated endocannabinoid levels in subcutaneous adipose tissue (SAT) of subjects with both obesity and type 2 diabetes (OBT2D), characterised by similar adiposity and whole body insulin resistance and lower plasma leptin levels as compared to non-diabetic obese subjects (OB).

Design and Methods

The levels of anandamide and 2-AG, and of the anandamide-related PPARα ligands, oleoylethanolamide (OEA) and palmitoylethanolamide (PEA), in the SAT obtained by abdominal needle biopsy in 10 OBT2D, 11 OB, and 8 non-diabetic normal-weight (NW) subjects, were measured by liquid chromatography-mass spectrometry. All subjects underwent a hyperinsulinaemic euglycaemic clamp.

Results

As compared to NW, anandamide, OEA and PEA levels in the SAT were 2-4.4-fold elevated (p < 0.05), and 2-AG levels 2.3-fold reduced (p < .05), in OBT2D but not in OB subjects. Anandamide, OEA and PEA correlated positively (p < .05) with SAT leptin mRNA and free fatty acid during hyperinsulinaemic clamp, and negatively with SAT LPL activity and plasma HDL-cholesterol, which were all specifically altered in OBT2D subjects.

Conclusions

The observed alterations emphasize, for the first time in humans, the potential different role and regulation of adipose tissue anandamide (and its congeners) and 2-AG in obesity and type 2 diabetes.
Eosinophilic pneumonia associated with bleomycin in a patient with mediastinal seminoma: a case report
Authors: Hapani Sanjaykumar , Chu David , Wu Shenhong
Published: April 2010

Introduction

Lung toxicities resulting from the chemotherapeutic agent bleomycin encompass a variety of pathological changes, including bronchiolitis obliterans organizing pneumonia, interstitial pneumonitis and progressive interstitial fibrosis. We report a rare case of eosinophilic pneumonia associated with bleomycin.

Case presentation

A 44-year-old Hispanic man with a primary mediastinal seminoma complicated by superior vena cava syndrome underwent treatment with four cycles of bleomycin, etoposide and cisplatin. He had a complete positive response to the chemotherapy. However, three months after treatment he presented with shortness of breath and severe hypoxemia associated with peripheral eosinophilia. Computed tomography showed bilateral diffuse interstitial infiltrates that were refractory to antibiotic treatment. A lung biopsy showed eosinophilic pneumonia. He was subsequently treated with high-dose prednisone, resulting in a complete resolution of his symptoms and lung infiltrates.

Conclusion

This case illustrates that eosinophilic pneumonia may be a late sequela of bleomycin toxicity, and may respond dramatically to steroid treatment.
Iron content of ferritin modulates its uptake by intestinal epithelium: implications for co-transport of prions
Authors: Bhupanapadu Sunkesula Solomon Raju , Luo Xiu , Das Dola , Singh Ajay , Singh Neena
Published: April 2010

The spread of Chronic Wasting Disease (CWD) in the deer and elk population has caused serious public health concerns due to its potential to infect farm animals and humans. Like other prion disorders such a sporadic Creutzfeldt-Jakob-disease of humans and Mad Cow Disease of cattle, CWD is caused by PrP-scrapie (PrPSc), a β-sheet rich isoform of a normal cell surface glycoprotein, the prion protein (PrPC). Since PrPSc is sufficient to cause infection and neurotoxicity if ingested by a susceptible host, it is important to understand the mechanism by which it crosses the stringent epithelial cell barrier of the small intestine. Possible mechanisms include co-transport with ferritin in ingested food and uptake by dendritic cells. Since ferritin is ubiquitously expressed and shares considerable homology among species, co-transport of PrPSc with ferritin can result in cross-species spread with deleterious consequences. We have used a combination of in vitro and in vivo models of intestinal epithelial cell barrier to understand the role of ferritin in mediating PrPSc uptake and transport. In this report, we demonstrate that PrPSc and ferritin from CWD affected deer and elk brains and scrapie from sheep resist degradation by digestive enzymes, and are transcytosed across a tight monolayer of human epithelial cells with significant efficiency. Likewise, ferritin from hamster brains is taken up by mouse intestinal epithelial cells in vivo, indicating that uptake of ferritin is not limited by species differences as described for prions. More importantly, the iron content of ferritin determines its efficiency of uptake and transport by Caco-2 cells and mouse models, providing insight into the mechanism(s) of ferritin and PrPSc uptake by intestinal epithelial cells.
Helicobacter pylori: a poor man's gut pathogen?
Authors: Khalifa Mohammed Mahdy , Sharaf Radwa Raed , Aziz Ramy Karam
Published: April 2010

Helicobacter pylori is one of the human pathogens with highest prevalence around the world; yet, its principal mode of transmission remains largely unknown. The role of H. pylori in gastric disease and cancer has not been established until the end of the 20th century. Since then, its epidemiology has been extensively studied, and an accruing body of literature suggests that not all humans are equally at risk of infection by this gut pathogen. Here, we briefly review the different epidemiological aspects of H. pylori infection with emphasis on those factors related to human poverty. The epidemiology of H. pylori infection is characterized by marked differences between developing and developed countries, notably among children. In addition, congruent lines of evidence point out to socioeconomic factors and living standards as main determinants of the age-dependent acquisition rate of H. pylori, and consequently its prevalence. These data are alarming in the light of the changing global climate and birth rate, which are expected to change the demography of our planet, putting more children at risk of H. pylori and its complications for years to come.
4G/5G polymorphism of PAI-1 gene is associated with multiple organ dysfunction and septic shock in pneumonia induced severe sepsis: prospective, observational, genetic study
Authors: Madách Krisztina , Aladzsity István , Szilágyi Ágnes , Fust George , Gál János , Pénzes István , Prohászka Zoltán
Published: April 2010

Introduction

Activation of inflammation and coagulation are closely related and mutually interdependent in sepsis. The acute-phase protein, plasminogen activator inhibitor-1 (PAI-1) is a key element in the inhibition of fibrinolysis. Elevated levels of PAI-1 have been related to worse outcome in pneumonia. We aimed to evaluate the effect of functionally relevant 4G/5G polymorphism of PAI-1 gene in pneumonia induced sepsis.

Methods

We enrolled 208 Caucasian patients with severe sepsis due to pneumonia admitted to an intensive care unit (ICU). Patients were followed up until ICU discharge or death. Clinical data were collected prospectively and the PAI-1 4G/5G polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism technique. Patients were stratified according to the occurrence of multiple organ dysfunction syndrome, septic shock or death.

Results

We found that carriers of the PAI-1 4G/4G and 4G/5G genotypes have a 2.74-fold higher risk for multiple organ dysfunction syndrome (odds ratio [OR] 95% confidence interval [CI] = 1.335 - 5.604; p = 0.006) and a 2.57-fold higher risk for septic shock (OR 95%CI = 1.180 - 5.615; p = 0.018) than 5G/5G carriers. The multivariate logistic regression analysis adjusted for independent predictors, such as age, nosocomial pneumonia and positive microbiological culture also supported that carriers of the 4G allele have a higher prevalence of multiple organ dysfunction syndrome (adjusted odds ratio [aOR] = 2.957; 95%CI = 1.306 -6.698; p = 0.009) and septic shock (aOR = 2.603; 95%CI = 1.137 - 5.959; p = 0.024). However, genotype and allele analyses have not shown any significant difference regarding mortality in models non-adjusted or adjusted for acute physiology and chronic health evaluation (APACHE) II. Patients bearing the 4G allele had higher disseminated intravascular coagulation (DIC) score at admission (p = 0.007) than 5G/5G carriers. Moreover, in 4G allele carriers the length of ICU stay of non-survivors was longer (p = 0.091), fewer ventilation-free days (p = 0.008) and days without septic shock (p = 0.095) were observed during the first 28 days.

Conclusions

In Caucasian patients with severe sepsis due to pneumonia carriers of the 4G allele of PAI-1 polymorphism have higher risk for multiple organ dysfunction syndrome and septic shock and in agreement they showed more fulminant disease progression based on continuous clinical variables.
Identification of Nesfatin-1 in Human and Murine Adipose Tissue: A Novel Depot-Specific Adipokine with Increased Levels in Obesity
Authors: Ramanjaneya Manjunath , Chen Jing , Brown James E. , Tripathi Gyanendra , Hallschmid Manfred , Patel Suketu , Kern Werner , Hillhouse Edward W. , Lehnert Hendrik , Tan Bee K. , Randeva Harpal S.
Published: April 2010

Nesfatin-1 is a recently identified anorexigenic peptide derived from its precursor protein, nonesterified fatty acid/nucleobindin 2 (NUCB2). Although the hypothalamus is pivotal for the maintenance of energy homeostasis, adipose tissue plays an important role in the integration of metabolic activity and energy balance by communicating with peripheral organs and the brain via adipokines. Currently no data exist on nesfatin-1 expression, regulation, and secretion in adipose tissue. We therefore investigated NUCB2/nesfatin-1 gene and protein expression in human and murine adipose tissue depots. Additionally, the effects of insulin, dexamethasone, and inflammatory cytokines and the impact of food deprivation and obesity on nesfatin-1 expression were studied by quantitative RT-PCR and Western blotting. We present data showing NUCB2 mRNA (P < 0.001), nesfatin-1 intracellular protein (P < 0.001), and secretion (P < 0.01) were significantly higher in sc adipose tissue compared with other depots. Also, nesfatin-1 protein expression was significantly increased in high-fat-fed mice (P < 0.01) and reduced under food deprivation (P < 0.01) compared with controls. Stimulation of sc adipose tissue explants with inflammatory cytokines (TNFα and IL-6), insulin, and dexamethasone resulted in a marked increase in intracellular nesfatin-1 levels. Furthermore, we present evidence that the secretion of nesfatin-1 into the culture media was dramatically increased during the differentiation of 3T3-L1 preadipocytes into adipocytes (P < 0.001) and after treatments with TNF-α, IL-6, insulin, and dexamethasone (P < 0.01). In addition, circulating nesfatin-1 levels were higher in high-fat-fed mice (P < 0.05) and showed positive correlation with body mass index in human. We report that nesfatin-1 is a novel depot specific adipokine preferentially produced by sc tissue, with obesity- and food deprivation-regulated expression.
Myocyte Androgen Receptors Increase Metabolic Rate and Improve Body Composition by Reducing Fat Mass
Authors: Fernando Shannon M. , Rao Pengcheng , Niel Lee , Chatterjee Diptendu , Stagljar Marijana , Monks D. Ashley
Published: April 2010

Testosterone and other androgens are thought to increase lean body mass and reduce fat body mass in men by activating the androgen receptor. However, the clinical potential of androgens for improving body composition is hampered by our limited understanding of the tissues and cells that promote such changes. Here we show that selective overexpression of androgen receptor in muscle cells (myocytes) of transgenic male rats both increases lean mass percentage and reduces fat mass. Similar changes in body composition are observed in human skeletal actin promoter driving expression of androgen receptor (HSA-AR) transgenic mice and result from acute testosterone treatment of transgenic female HSA-AR rats. These shifts in body composition in HSA-AR transgenic male rats are associated with hypertrophy of type IIb myofibers and decreased size of adipocytes. Metabolic analyses of transgenic males show higher activity of mitochondrial enzymes in skeletal muscle and increased O2 consumption by the rats. These results indicate that androgen signaling in myocytes not only increases muscle mass but also reduces fat body mass, likely via increases in oxidative metabolism.
Role of the TSC1-TSC2 Complex in the Integration of Insulin and Glucose Signaling Involved in Pancreatic β-Cell Proliferation
Authors: Bartolomé Alberto , Guillén Carlos , Benito Manuel
Published: April 2010

Tuberous sclerosis complex proteins 1–2 (TSC1-TSC2) complex integrates both nutrient and hormonal signaling and is a critical negative regulator of mammalian target of rapamycin (mTOR) complex 1. The use of different β-cell lines expressing or not the insulin receptor (IR+/+ and IR−/−) or with a reconstituted expression of IR isoform A or B (Rec A and Rec B) revealed that both phosphatidylinositol 3-kinase/Akt/TSC/mTOR complex 1 and MAPK kinase/ERK pathways mediate insulin signaling in IR+/+-, IRA-, or IRB-expressing cells. However, glucose signaling was mediated by MAPK kinase/ERK and AMP-activated protein kinase pathways as assessed in IR−/− cells. The effect of insulin on Akt phosphorylation was completely inhibited by the use of the phosphatidylinositol 3-kinase inhibitor wortmannin in IR+/+ and Rec B cells, a partial inhibitory effect being observed in Rec A cell line. The knockdown of TSC2 expression up-regulated the downstream basal phosphorylation of 70-kDa ribosomal protein S6 kinase (p70S6K) and mTOR. More importantly, upregulation of p70S6K signaling impaired insulin-stimulated phosphorylation of Akt Ser473 and p70S6K in IR+/+ and Rec B but not in Rec A cell lines. In fact, insulin receptor substrate-1 Ser307 phosphorylation signal in Rec B was stronger than in Rec A cell line during insulin action. Rec A cells induced a higher proliferation rate compared with Rec B or IR+/+ during serum stimulation. Thus, we propose that the regulation of TSC2 phosphorylation by insulin or glucose independently integrates β-cell proliferation signaling, the relative expression of IRA or IRB isoforms in pancreatic β cells playing a major role.