bluuebunny/crossref_metadata_2025 huggingface dataset:
Mobilities II
Authors: Cresswell Tim
Published: September 2011

This second report on mobilities considers some key themes in mobilities research by (mostly) geographers over the last two years or so. Following on from some of the themes outlined in the first report, this report explores accounts of historical geographies of mobility in order to put claims to ‘newness’ in perspective. Second, it surveys how mobility research has influenced methodology focusing, in particular, on ‘mobile ethnography’. Third, the report looks at the blossoming arena or research on the forms of waiting, stillness and stuckness that have become an important component of our understanding of mobility. The conclusion reflects on the continuing importance of the politics of mobility and urges greater consideration of the mobility of ideas alongside people and things.
Creativity and Innovation
Authors: Stein Stanley M. , Harper Thomas L.
Published: September 2011

Planning theory based on Habermassian ideas of rationality has been criticized as inhibiting creativity, rendering it incapable of dealing with pressing social problems. This critique implicitly assumes a traditional view of communication and language, one not matching actual life experience. Our “pragmatic-communicative” approach—“dialogical planning”—draws on a nontraditional view: communication as interpretation. This encourages creativity and facilitates innovation through flexibility and divergence, yet seeks convergence by connecting new ideas to our web of existing concepts and beliefs. Planners can better stimulate creativity and innovation if they understand and employ this pragmatic view of communication and language.
Pallidal Deep-Brain Stimulation Associated With Complete Remission of Self-injurious Behaviors in a Patient With Lesch-Nyhan Syndrome
Authors: Deon Laura L. , Kalichman Miriam A. , Booth Cynthia L. , Slavin Konstantin V. , Gaebler-Spira Deborah J.
Published: September 2011

The purpose of this case report is to review the management of a boy with Lesch-Nyhan syndrome with deep-brain stimulation who had remission of self-injurious behaviors as a result. This patient was treated with intrathecal baclofen and, later, with deep-brain stimulation to reduce hypertonia. Goals were to improve wheelchair positioning for school attendance and to reduce the use of restraints for comfort. Intrathecal baclofen was implanted twice and decreased the hypertonia, but both were explanted because of infection. Deep-brain stimulation was initiated 2.5 years ago, and since that time, comfort and function have improved and caregiver burden has decreased. Improvements in dystonia with deep-brain stimulation have also occurred, and self-injurious behaviors have resolved.
Long-Term Outcome in Children With Intractable Epilepsy Showing Bilateral Diffuse Cortical Glucose Hypometabolism Pattern on Positron Emission Tomography
Authors: Shandal Varun , Veenstra Amy L. , Behen Michael , Sundaram Senthil , Chugani Harry
Published: September 2011

The objective of this study is to determine the long-term outcome of children with intractable epilepsy who have diffuse cortical hypometabolism on 2-deoxy-2-( 18 F)fluoro-D-glucose positron emission tomography (FDG-PET) scans. Seventeen children with intractable epilepsy showing bilateral, diffuse cortical hypometabolism on FDG-PET were followed up through telephone interview from 1 year 4 months to 11 years 4 months (mean: 5 years 7 months ± 2 years 1 month) after their PET scans. One child succumbed to Sanfilippo disease at age 20 years. Only 2 children were seizure free. Fifty percent had walking difficulties, 56.25% were not toilet trained, all had speech difficulties, 43.75% had behavioral problems, 37.5% had poor eye contact, 75% had socialization difficulties, and 87.5% attended special schools. Three children were found to have genetic causes, including a 4-MB deletion of the mitochondrial genome, MECP2 duplication, and Lafora disease. In conclusion, the long-term outcome in this patient population is poor, and they tend to suffer from genetic/neurodegenerative diseases.
Cerebellar Gliomatosis in a Toddler
Authors: de Chadarévian Jean-Pierre , Legido Agustin , Halligan Gregory E. , Faerber Eric N. , Piatt Joseph H. , Morrissette Jennifer D. , Ara Jahan , Grant Mitzie L. , Katsetos Christos D.
Published: September 2011

Gliomatosis confined to the cerebellum is most unusual. We report such a case in a 20-month-old male who presented with unsteadiness. Magnetic resonance imaging revealed a diffuse area of abnormal signal intensity within both cerebellar hemispheres, which did not enhance after contrast administration. The patient underwent a biopsy, which revealed a diffuse glioma infiltrating the cerebellum. Overall, the tumor cells had oligodendroglioma-like features and exhibited only focal vimentin immunoreactivity. They were negative for glial fibrillary acidic protein, synaptophysin, βIII-tubulin, and neurofilament protein. Immunofluorescence, performed on primary biopsy explants maintained in cell culture without exposure to growth factors or differentiation-promoting agents, revealed widespread nestin immunoreactivity and immunolabeling of occasional cells with antibodies to platelet-derived growth factor-α and O1/O4, markers of oligodendrocyte precursor-cells and immature oligodendrocytes, respectively. Fluorescent in situ hybridization performed on explants, touch preparations, and paraffin sections failed to reveal loss of heterozygosity for either 1p36 or 19q13. The patient was treated with temozolomide and remains stable, albeit with residual quiescent tumor, more than 3 years after surgery. This report calls attention to an unusual presentation of gliomatosis confined to the cerebellum of a toddler and addresses salient aspects of clinical and radiological differential diagnosis, as well as therapeutic challenges encountered.
Drooling
Authors: Nordgarden Hilde , Østerhus Ingvild , Møystad Anne , Åsten Pamela , Johnsen Ulf L-H , Storhaug Kari , Loven Jens Øyvind
Published: September 2011

There are several treatment options available for drooling; botulinum toxin injections into the major salivary glands are one. There is no consensus as to how many and which glands should be injected. A research project on this topic was terminated because of adverse effects. Individual results and the adverse effects are described and discussed in this article. Six individuals with cerebral palsy were randomly allocated to 2 treatment groups, with five individuals receiving ultrasound-guided injections to parotid and submandibular glands and one receiving injections to the submandibular glands only. Reduction of observed drooling was registered in 3, while 4 patients reported subjective improvement (Visual Analog Scale). Two participants reported adverse effects, including dysphagia, dysarthria, and increased salivary viscosity. Injections with botulinum toxin can be a useful treatment option but there is a risk of adverse effects. Multidisciplinary evaluation and informed discussions with patients/caregivers are important factors in the decision-making process.
Acculturation, Enculturation, Ethnic Identity, and Conscientiousness as Predictors of Latino Boys’ and Girls’ Career Decision Self-Efficacy
Authors: Ojeda Lizette , Piña-Watson Brandy , Castillo Linda G. , Castillo Rosalinda , Khan Noshaba , Leigh Jennifer
Published: September 2011

This study examined the role of culture and personality on the career decision self-efficacy of 338 Latino seventh-grade public middle school students. Specifically, we examined the role of acculturation, enculturation, ethnic identity, and conscientiousness on career decision self-efficacy. Findings indicated Latina girls were more acculturated and enculturated than Latino boys. For Latino boys and Latina girls, respectively, 34% and 25% of the variance in career decision self-efficacy was accounted for by the predictors. Acculturation, ethnic identity, and conscientiousness significantly predicted career decision self-efficacy for girls. For boys, however, only ethnic identity and conscientiousness were unique significant predictors. As expected, enculturation did not significantly predict career decision self-efficacy for either boys or girls. An interaction effect between acculturation and enculturation did not significantly predict career decision self-efficacy for boys or girls. Implications for practice and future research are discussed.
Counterdependence at Work
Authors: Gianakos Irene
Published: September 2011

Although styles of self-reliance theoretically arise from early attachments, other research suggests the influence of attachment operates indirectly through internalized but modifiable beliefs about the self and others. The present study examined the relative contribution of parental attachment, social support perceptions, control beliefs, and self-monitoring in understanding workplace counterdependence, a problematic style of self-reliance reflective of avoidant attachments. Participants were 184 undergraduate students (mean age = 23.2 years) who worked an average of 29.2 hours per week. As expected, the regression analysis identified perceptions about support, explanatory beliefs, and self-monitoring skills as significant predictors of counterdependence, whereas parental attachment factors were not; collectively these significant predictors explained 41.5% of the variance. The results suggest that difficulties with workplace counterdependence may be ameliorated by interventions that target workers' perceived deficiencies in the skill-linked predictors identified in this study.
Prevalence and Incidence Rates of Dementia and Cognitive Impairment No Dementia in the Mexican Population
Authors: Mejia-Arango Silvia , Gutierrez Luis Miguel
Published: September 2011

Objective: To estimate the prevalence and incidence of dementia and cognitive impairment without dementia (CIND) in the Mexican population. Method: The MHAS study is a prospective panel study of health and aging in Mexico with 7,000 elders that represent eight million participants nationally. Using measurements of cognition and activities of daily living of dementia cases and CIND were identified at baseline and follow-up. Overall incidence rates and specific rates for sex, age, and education were calculated. Results: Prevalence was 6.1% and 28.7% for dementia and CIND, respectively. Incidence rates were 27.3 per 1,000 person-years for dementia and 223 per 1,000 persons-year for CIND. Rates of dementia and CIND increased with advancing age and decreased with higher educational level; sex had a differential effect depending on the age strata. Hypertension, diabetes, and depression were risk factors for dementia but not for CIND. Discussion: These data provide estimates of prevalence and incidence of dementia and cognitive impairment in the Mexican population for projection of future burden.
The Metabolic Syndrome, Biomarkers, and the Acculturation–Health Relationship Among Older Mexican Americans
Authors: González Hector M. , Tarraf Wassim , Haan Mary N.
Published: September 2011

Objective: To examine the acculturation–health relationship using metabolic syndrome biomarkers. Method: Cross-sectional sample data. Participants: 1,789 Mexican Americans (60 years and older) from northern California. Main Outcome: Biomarkers (waist circumference, blood pressure, fasting plasma glucose, triglycerides, and high-density lipids) were used to construct the metabolic syndrome indicator using American Heart Association criteria. Main Predictor: Acculturation Rating Scale for Mexican Americans-II scores. Results: Higher acculturation scores were associated with a significantly lower risk for the metabolic syndrome for foreign-born, but not U.S.-born, Mexican Americans. Conclusion: Immigrant health advantages over U.S.-born Mexican Americans are not evident in older adulthood. Higher acculturation was associated with lowered metabolic syndrome risk among older foreign-born Mexican Americans. This suggests that the prevailing acculturative stress hypothesis may not apply to the health of older adults and that any negative relationship between acculturation and health found in younger adults may yield to different developmental health influences in later adulthood.
Links Between Childhood and Adult Social Circumstances and Obesity and Hypertension in the Mexican Population
Authors: Beltrán-Sánchez Hiram , Crimmins Eileen M. , Teruel Graciela M. , Thomas Duncan
Published: September 2011

Objectives: This study examines links between early life circumstances and adult socioeconomic status and obesity and hypertension in the adult Mexican population. Method: We use data from the Mexican Family Life Survey (MxFLS) collected in 2002 for people aged 20 or older ( N = 14,280). Results: We found that men with low education and women with more education have significantly lower obesity. Women with higher education also have significantly less hypertension. Obesity triples the likelihood of hypertension among both men and women. Better childhood experiences are associated with less hypertension among women, but more hypertension among men in rural areas. Discussion: Recent changes in income, nutrition, and infection in Mexico may be responsible for the observed high prevalence of overweight and obesity and the extremely high odds of hypertension among obese young adults.
Mothers’ Process of Decision Making for Gastrostomy Placement
Authors: Brotherton Ailsa , Abbott Janice
Published: September 2011

In this article we present the findings of an exploration of mothers’ discourses on decision making for gastrostomy placement for their child. Exploring in-depth interviews of a purposive sample, we analyzed the mothers’ discourses of the decision-making process to understand how their experiences of the process influenced their subsequent constructions of decision making. Mothers negotiated decision making by reflecting on their personal experiences of feeding their child, either orally or via a tube, and interwove their background experiences with the communications from members of the health care team until a decision was reached. Decision making was often fraught with difficulty, resulting in anxiety and guilt. Experiences of decision making ranged from perceived coercion to true choice, which encompasses a truly child-centered decision. The resulting impact of the decision-making process on the mothers was profound. We conclude with an exploration of the implications for clinical practice and describe how health care professionals can support mothers to ensure that decision-making processes for gastrostomy placement in children are significantly improved.
Canadian Care Providers’ and Pregnant Women’s Approaches to Managing Birth
Authors: Hall Wendy A. , Tomkinson Jocelyn , Klein Michael C.
Published: September 2011

We employed grounded theory to explain how Canadian pregnant women and care providers manage birth. The sample comprised 9 pregnant women and 56 intrapartum care providers (family doctors, midwives, nurses, obstetricians, and doulas [individuals providing labor support]). We collected data from 2008 to 2009, using focus groups that included care providers and pregnant women. Using concurrent data collection and analysis, we generated the core category: minimizing risk while maximizing integrity. Women and providers used strategies to minimize risk and maximize integrity, which included accepting or resisting recommendations for surveillance and recommendations for interventions, and plotting courses vs. letting events unfold. Strategies were influenced by evidence, relationships, and local health cultures, and led to feelings of weakness or strength, confidence or uncertainty, and differing power- and responsibility-sharing arrangements. The findings highlight difficulties resisting surveillance and interventions in a risk-adverse culture, and the need for attention to processes of giving birth.
Explaining Ups and Downs in the Public’s Environmental Concern in Sweden
Authors: Harring Niklas , Jagers Sverker C. , Martinsson Johan
Published: September 2011

In this article, the authors search for explanations to ups and downs in the Swedish public’s environmental concern since the 1980s. In line with previous research, this study examines the effects of economic cycles and media coverage. In addition, the authors hypothesize that the economy will affect environmental concern less over time because of the entry of ecological modernization into elite discourse. Using time series regression analysis and a unique data set, we study Swedish public opinion during more than 20 years. Economic cycles affect the public’s environmental concern but to a diminishing degree. Public environmental concern is also affected by the amount of media coverage. In accordance with earlier observations, it is concluded that both the economy and media content have an independent effect on public environmental concern. However, the previously observed conflict between economic cycles and public environmental concern is weakened, potentially because of the elite group embracement of an ecological modernization discourse.
Physical activity patterns in Greenland: A country in transition
Authors: Dahl-Petersen Inger K. , Jørgensen Marit E. , Bjerregaard Peter
Published: September 2011

Aims: To examine differences in physical activity patterns among Inuit in Greenland in relation to social transition. The Inuit in Greenland are an indigenous population in the circumpolar north who are experiencing rapid social transition. Methods: Physical activity patterns were assessed by the International Physical Activity Questionnaire (long version). The population was divided into six groups according to different stages of social change, measured on the basis of education, current residence and occupation. Data were collected in a country-wide cross-sectional population survey among adult Inuit in Greenland from 2005 to 2009. Results: Men with long vocational or academic education living in towns (latest stage of social change) spent significantly less time on occupational physical activity (p = 0.001) compared with hunters and fishermen in villages (earliest stage of social change) (trend test p = 0.01). Women in the latest stage of change spent significantly less time on domestic physical activity (p < 0.001) (trend test p = 0.06) compared with women in the earliest stage of social change. This was also found for physical activity during transportation (p = 0.02 and p = 0.01 for men and women, respectively). No significant difference was found for leisuretime physical activity. Men and women in the latest stage of social change spent more time on sedentary activity (p < 0.001). Conclusions: Differences in physical activity patterns among Inuit in Greenland included decreasing time spent on domestic and occupational physical activity and increasing time spent on sedentary activities along with social change. Knowledge of changes in physical activity patterns in relation to social transition is important in prevention of obesity, type 2 diabetes and lifestyle diseases.
Walk a mile in my shoes: Life as a mother of a child with Asperger's Syndrome
Authors: Gill Jessica , Liamputtong Pranee
Published: September 2011

In this study, we explore the experiences of mothers with children who have Asperger's Syndrome. A qualitative inquiry was adopted in our study. Fifteen women participated in in-depth interviews, six of whom produced a solicited diary that offered a more in-depth insight into their lives. Thematic analysis was used to examine themes from the transcripts and diary entries. A number of themes emerged from the interview and diary data. First, the women articulated their perceptions of motherhood which included perceptions of an ideal mother, the mother role, the physical and emotional health of mothers and the mask of motherhood. Second, the data revealed the moral career of motherhood which discussed mother blame as a main issue. The findings of this study have implications for health and social care provided to women who are mothers of children with Asperger's Syndrome.
Regulation of macrophage migration by a novel plasminogen receptor Plg-RKT
Authors: Lighvani Shahrzad , Baik Nagyung , Diggs Jenna E. , Khaldoyanidi Sophia , Parmer Robert J. , Miles Lindsey A.
Published: September 2011

Localization of plasmin on macrophages and activation of pro–MMP-9 play key roles in macrophage recruitment in the inflammatory response. These functions are promoted by plasminogen receptors exposing C-terminal basic residues on the macrophage surface. Recently, we identified a novel transmembrane plasminogen receptor, Plg-RKT, which exposes a C-terminal lysine on the cell surface. In the present study, we investigated the role of Plg-RKT in macrophage invasion, chemotactic migration, and recruitment. Plg-RKT was prominently expressed in membranes of human peripheral blood monocytes and monocytoid cells. Plasminogen activation by urokinase-type plasminogen activator (uPA) was markedly inhibited (by 39%) by treatment with anti–Plg-RKT mAb. Treatment of monocytes with anti–Plg-RKT mAb substantially inhibited invasion through the representative matrix, Matrigel, in response to MCP-1 (by 54% compared with isotype control). Furthermore, chemotactic migration was also inhibited by treatment with anti–Plg-RKT mAb (by 64%). In a mouse model of thioglycollate-induced peritonitis, anti–Plg-RKT mAb markedly inhibited macrophage recruitment (by 58%), concomitant with a reduction in pro–MMP-9 activation in the inflamed peritoneum. Treatment with anti–Plg-RKT mAb did not further reduce the low level of macrophage recruitment in plasminogen-null mice. We conclude that Plg-RKT plays a key role in the plasminogen-dependent regulation of macrophage invasion, chemotactic migration, and recruitment in the inflammatory response.
Iron regulatory protein-1 and -2: transcriptome-wide definition of binding mRNAs and shaping of the cellular proteome by iron regulatory proteins
Authors: Sanchez Mayka , Galy Bruno , Schwanhaeusser Bjoern , Blake Jonathon , Bähr-Ivacevic Tomi , Benes Vladimir , Selbach Matthias , Muckenthaler Martina U. , Hentze Matthias W.
Published: September 2011

Iron regulatory proteins (IRPs) 1 and 2 are RNA-binding proteins that control cellular iron metabolism by binding to conserved RNA motifs called iron-responsive elements (IREs). The currently known IRP-binding mRNAs encode proteins involved in iron uptake, storage, and release as well as heme synthesis. To systematically define the IRE/IRP regulatory network on a transcriptome-wide scale, IRP1/IRE and IRP2/IRE messenger ribonucleoprotein complexes were immunoselected, and the mRNA composition was determined using microarrays. We identify 35 novel mRNAs that bind both IRP1 and IRP2, and we also report for the first time cellular mRNAs with exclusive specificity for IRP1 or IRP2. To further explore cellular iron metabolism at a system-wide level, we undertook proteomic analysis by pulsed stable isotope labeling by amino acids in cell culture in an iron-modulated mouse hepatic cell line and in bone marrow-derived macrophages from IRP1- and IRP2-deficient mice. This work investigates cellular iron metabolism in unprecedented depth and defines a wide network of mRNAs and proteins with iron-dependent regulation, IRP-dependent regulation, or both.
Coevolution of amino acid residues in the key photosynthetic enzyme Rubisco
Authors: Wang Mingcong , Kapralov Maxim V , Anisimova Maria
Published: September 2011

Background

One of the key forces shaping proteins is coevolution of amino acid residues. Knowing which residues coevolve in a particular protein may facilitate our understanding of protein evolution, structure and function, and help to identify substitutions that may lead to desired changes in enzyme kinetics. Rubisco, the most abundant enzyme in biosphere, plays an essential role in the process of carbon fixation through photosynthesis, thus facilitating life on Earth. This makes Rubisco an important model system for studying the dynamics of protein fitness optimization on the evolutionary landscape. In this study we investigated the selective and coevolutionary forces acting on large subunit of land plants Rubisco using Markov models of codon substitution and clustering approaches applied to amino acid substitution histories.

Results

We found that both selection and coevolution shape Rubisco, and that positively selected and coevolving residues have their specifically favored amino acid composition and pairing preference. The mapping of these residues on the known Rubisco tertiary structures showed that the coevolving residues tend to be in closer proximity with each other compared to the background, while positively selected residues tend to be further away from each other. This study also reveals that the residues under positive selection or coevolutionary force are located within functionally important regions and that some residues are targets of both positive selection and coevolution at the same time.

Conclusion

Our results demonstrate that coevolution of residues is common in Rubisco of land plants and that there is an overlap between coevolving and positively selected residues. Knowledge of which Rubisco residues are coevolving and positively selected could be used for further work on structural modeling and identification of substitutions that may be changed in order to improve efficiency of this important enzyme in crops.
Transcriptome profiling of immune responses to cardiomyopathy syndrome (CMS) in Atlantic salmon
Authors: Timmerhaus Gerrit , Krasnov Aleksei , Nilsen Pål , Alarcon Marta , Afanasyev Sergey , Rode Marit , Takle Harald , Jørgensen Sven Martin
Published: September 2011

Background

Cardiomyopathy syndrome (CMS) is a disease associated with severe myocarditis primarily in adult farmed Atlantic salmon (Salmo salarL.), caused by a double-stranded RNA virus named piscine myocarditis virus (PMCV) with structural similarities to theTotiviridaefamily. Here we present the first characterisation of host immune responses to CMS assessed by microarray transcriptome profiling.

Results

Unvaccinated farmed Atlantic salmon post-smolts were infected by intraperitoneal injection of PMCV and developed cardiac pathology consistent with CMS. From analysis of heart samples at several time points and different tissues at early and clinical stages by oligonucleotide microarrays (SIQ2.0 chip), six gene sets representing a broad range of immune responses were identified, showing significant temporal and spatial regulation. Histopathological examination of cardiac tissue showed myocardial lesions from 6 weeks post infection (wpi) that peaked at 8-9 wpi and was followed by a recovery. Viral RNA was detected in all organs from 4 wpi suggesting a broad tissue tropism. High correlation between viral load and cardiac histopathology score suggested that cytopathic effect of infection was a major determinant of the myocardial changes. Strong and systemic induction of antiviral and IFN-dependent genes from 2 wpi that levelled off during infection, was followed by a biphasic activation of pathways for B cells and MHC antigen presentation, both peaking at clinical pathology. This was preceded by a distinct cardiac activation of complement at 6 wpi, suggesting a complement-dependent activation of humoral Ab-responses. Peak of cardiac pathology and viral load coincided with cardiac-specific upregulation of T cell response genes and splenic induction of complement genes. Preceding the reduction in viral load and pathology, these responses were probably important for viral clearance and recovery.

Conclusions

By comparative analysis of gene expression, histology and viral load, the temporal and spatial regulation of immune responses were characterised and novel immune genes identified, ultimately leading to a more complete understanding of host-virus responses and pathology and protection in Atlantic salmon during CMS.
The Aspergillus giganteus antifungal protein AFPNN5353activates the cell wall integrity pathway and perturbs calcium homeostasis
Authors: Binder Ulrike , Bencina Mojca , Eigentler Andrea , Meyer Vera , Marx Florentine
Published: September 2011

Background

The antifungal protein AFPNN5353 is a defensin-like protein of Aspergillus giganteus. It belongs to a group of secretory proteins with low molecular mass, cationic character and a high content of cysteine residues. The protein inhibits the germination and growth of filamentous ascomycetes, including important human and plant pathogens and the model organsims Aspergillus nidulans and Aspergillus niger.

Results

We determined an AFPNN5353 hypersensitive phenotype of non-functional A. nidulans mutants in the protein kinase C (Pkc)/mitogen-activated protein kinase (Mpk) signalling pathway and the induction of the α-glucan synthase A (agsA) promoter in a transgenic A. niger strain which point at the activation of the cell wall integrity pathway (CWIP) and the remodelling of the cell wall in response to AFPNN5353. The activation of the CWIP by AFPNN5353, however, operates independently from RhoA which is the central regulator of CWIP signal transduction in fungi.

Furthermore, we provide evidence that calcium (Ca2+) signalling plays an important role in the mechanistic function of this antifungal protein. AFPNN5353 increased about 2-fold the cytosolic free Ca2+ ([Ca2+]c) of a transgenic A. niger strain expressing codon optimized aequorin. Supplementation of the growth medium with CaCl2 counteracted AFPNN5353 toxicity, ameliorated the perturbation of the [Ca2+]c resting level and prevented protein uptake into Aspergillus sp. cells.

Conclusions

The present study contributes new insights into the molecular mechanisms of action of the A. giganteus antifungal protein AFPNN5353. We identified its antifungal activity, initiated the investigation of pathways that determine protein toxicity, namely the CWIP and the Ca2+ signalling cascade, and studied in detail the cellular uptake mechanism in sensitive target fungi. This knowledge contributes to define new potential targets for the development of novel antifungal strategies to prevent and combat infections of filamentous fungi which have severe negative impact in medicine and agriculture.
Extracellular ATP activates NFAT-dependent gene expression in neuronal PC12 cells via P2X receptors
Authors: Prasai Prabin , Stefos Georgios C , Becker Walter
Published: September 2011

Background

Treatment of neuronal PC12 cells with ATP induces depolarisation and increases intracellular calcium levels via purinergic receptors. In many cell types, sustained elevation of intracellular calcium levels cause changes in gene expression via activation of the transcription factor NFAT (nuclear factor of activated T cells). We have therefore characterised the signalling pathway by which ATP regulates NFAT-dependent gene expression in PC12 cells.

Results

The activation of NFAT transcriptional activity by extracellular ATP was characterised with the help of reporter gene assays. Treatment of PC12 cells with ATP elicited a dose-dependent increase in luciferase activity (EC50 = 78 μM). UTP, 4-benzoylbenzoyl ATP and α,β-methylene ATP did not mimic the effect of ATP, which was abolished by treatment with the P2X receptor antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS). This pharmacological characterisation provides evidence for a critical role of ionotropic P2X receptors. Blockade of L-type voltage-dependent calcium channels by nifedipine reduced the response of NFAT to ATP, indicating that a depolarisation-mediated calcium influx was required for maximal NFAT activation. Inhibition of store-operated calcium entry by the pyrazole derivative BTP2 also diminished ATP-dependent NFAT activation. Furthermore, ATP-induced NFAT activation was associated with the activation of the mitogen-activated protein kinases ERK1/2. Finally, treatment with ATP increased the levels of the NFAT target transcripts, RCAN1-4 (regulator of calcineurin) and BDNF (brain derived neurotrophic factor).

Conclusion

The present data show that ATP induces NFAT-dependent changes in gene expression in PC12 cells by acting on P2X receptors. Maximal NFAT activation depends on both depolarisation-induced calcium influx and store-operated calcium entry and requires the activity of the protein phosphatase calcineurin and the mitogen-activated protein kinase cascade.
Directed PCR-free engineering of highly repetitive DNA sequences
Authors: Scior Annika , Preissler Steffen , Koch Miriam , Deuerling Elke
Published: September 2011

Background

Highly repetitive nucleotide sequences are commonly found in nature e.g. in telomeres, microsatellite DNA, polyadenine (poly(A)) tails of eukaryotic messenger RNA as well as in several inherited human disorders linked to trinucleotide repeat expansions in the genome. Therefore, studying repetitive sequences is of biological, biotechnological and medical relevance. However, cloning of such repetitive DNA sequences is challenging because specific PCR-based amplification is hampered by the lack of unique primer binding sites resulting in unspecific products.

Results

For the PCR-free generation of repetitive DNA sequences we used antiparallel oligonucleotides flanked by restriction sites of Type IIS endonucleases. The arrangement of recognition sites allowed for stepwise and seamless elongation of repetitive sequences. This facilitated the assembly of repetitive DNA segments and open reading frames encoding polypeptides with periodic amino acid sequences of any desired length. By this strategy we cloned a series of polyglutamine encoding sequences as well as highly repetitive polyadenine tracts. Such repetitive sequences can be used for diverse biotechnological applications. As an example, the polyglutamine sequences were expressed as His6-SUMO fusion proteins in Escherichia coli cells to study their aggregation behavior in vitro. The His6-SUMO moiety enabled affinity purification of the polyglutamine proteins, increased their solubility, and allowed controlled induction of the aggregation process. We successfully purified the fusions proteins and provide an example for their applicability in filter retardation assays.

Conclusion

Our seamless cloning strategy is PCR-free and allows the directed and efficient generation of highly repetitive DNA sequences of defined lengths by simple standard cloning procedures.
Adherence to prescribed artemisinin-based combination therapy in Garissa and Bunyala districts, Kenya
Authors: Lawford Harriet , Zurovac Dejan , O'Reilly Laura , Hoibak Sarah , Cowley Alice , Munga Stephen , Vulule John , Juma Elizabeth , Snow Robert W , Allan Richard
Published: September 2011

Background

Following the development of resistance to anti-malarial mono-therapies, malaria endemic countries in Africa now use artemisinin-based combination therapy (ACT) as recommended first-line treatment for uncomplicated malaria. Patients' adherence to ACT is an important factor to ensure treatment efficacy, as well as to reduce the likelihood of parasite resistance to these drugs. This study reports adherence to a specific ACT, artemether-lumefantrine (AL), under conditions of routine clinical practice in Kenya.

Method

The study was undertaken in Garissa and Bunyala districts among outpatients of five government health facilities. Patients treated with AL were visited at home four days after having been prescribed the drug. Respondents (patients ≥ 15 years and caregivers of patients < 15 years) were interviewed using a standardized questionnaire, AL blister packs were physically inspected and the adherence status of patients was then recorded. Multivariate logistic regression modelling was used to determine predictors of adherence.

Results

Of the 918 patients included in the study, 588 (64.1%) were 'probably adherent', 291 (31.7%) were 'definitely non-adherent' and 39 (4.2%) were 'probably non-adherent'. Six factors were found to be significant predictors of adherence: patient knowledge of the ACT dosing regimen (odds ratio (OR) = 1.76; 95% CI = 1.32-2.35), patient age (OR = 1.65; 95% CI = 1.02-1.85), respondent age (OR = 1.37; 95% CI = 1.10-2.48), whether a respondent had seen AL before (OR = 1.46; 95% CI = 1.08-1.98), whether a patient had reported dislikes to AL (OR = 0.62 95% CI = 0.47-0.82) and whether a respondent had waited more than 24 hours to seek treatment (OR = 0.73; 95% CI = 0.54-0.99).

Conclusion

Overall, adherence to AL was found to be low in both Garissa and Bunyala districts, with patient knowledge of the AL dosing regimen found to be the strongest predictor of adherence. Interventions aimed at increasing community awareness of the AL dosing regimen, use of child friendly formulations and improving health workers' prescribing practices are likely to ensure higher adherence to AL and eventual treatment success.
BRAF and RAS oncogenes regulate Rho GTPase pathways to mediate migration and invasion properties in human colon cancer cells: a comparative study
Authors: Makrodouli Eleni , Oikonomou Eftychia , Koc Michal , Andera Ladislav , Sasazuki Takehiko , Shirasawa Senji , Pintzas Alexander
Published: September 2011

Background

Colorectal cancer is a common disease that involves genetic alterations, such as inactivation of tumour suppressor genes and activation of oncogenes. Among them are RAS and BRAF mutations, which rarely coexist in the same tumour. Individual members of the Rho (Ras homology) GTPases contribute with distinct roles in tumour cell morphology, invasion and metastasis. The aim of this study is to dissect cell migration and invasion pathways that are utilised by BRAFV600Eas compared to KRASG12Vand HRASG12Voncoproteins. In particular, the role of RhoA (Ras homolog gene family, member A), Rac1 (Ras-related C3 botulinum toxin substrate 1) and Cdc42 (cell division cycle 42) in cancer progression induced by each of the three oncogenes is described.

Methods

Colon adenocarcinoma cells with endogenous as well as ectopically expressed or silenced oncogenic mutations of BRAFV600E, KRASG12Vand HRASG12Vwere employed. Signalling pathways and Rho GTPases were inhibited with specific kinase inhibitors and siRNAs. Cell motility and invasion properties were correlated with cytoskeletal properties and Rho GTPase activities.

Results

Evidence presented here indicate that BRAFV600Esignificantly induces cell migration and invasion propertiesin vitroin colon cancer cells, at least in part through activation of RhoA GTPase. The relationship established between BRAFV600Eand RhoA activation is mediated by the MEK-ERK pathway. In parallel, KRASG12Venhances the ability of colon adenocarcinoma cells Caco-2 to migrate and invade through filopodia formation and PI3K-dependent Cdc42 activation. Ultimately increased cell migration and invasion, mediated by Rac1, along with the mesenchymal morphology obtained through the Epithelial-Mesenchymal Transition (EMT) were the main characteristics rendered by HRASG12Vin Caco-2 cells. Moreover, BRAF and KRAS oncogenes are shown to cooperate with the TGFβ-1 pathway to provide cells with additional transforming properties.

Conclusion

This study discriminates oncogene-specific cell migration and invasion pathways mediated by Rho GTPases in colon cancer cells and reveals potential new oncogene-specific characteristics for targeted therapeutics.
A palatable hyperlipidic diet causes obesity and affects brain glucose metabolism in rats
Authors: Estadella Debora , Oyama Lila M , Bueno Allain A , Habitante Carlos A , Souza Gabriel I , Ribeiro Eliane B , Motoyama Caio SM , Oller do Nascimento Claudia M
Published: September 2011

Background

We have previously shown that either the continuous intake of a palatable hyperlipidic diet (H) or the alternation of chow (C) and an H diet (CH regimen) induced obesity in rats. Here, we investigated whether the time of the start and duration of these feeding regimens are relevant and whether they affect brain glucose metabolism.

Methods

Male Wistar rats received C, H, or CH diets during various periods of their life spans: days 30-60, days 30-90, or days 60-90. Experiments were performed the 60th or the 90th day of life. Rats were killed by decapitation. The glucose, insulin, leptin plasma concentration, and lipid content of the carcasses were determined. The brain was sliced and incubated with or without insulin for the analysis of glucose uptake, oxidation, and the conversion of [1-14C]-glucose to lipids.

Results

The relative carcass lipid content increased in all of the H and CH groups, and the H30-60 and H30-90 groups had the highest levels. Groups H30-60, H30-90, CH30-60, and CH30-90 exhibited a higher serum glucose level. Serum leptin increased in all H groups and in the CH60-90 and CH30-90 groups. Serum insulin was elevated in the H30-60, H60-90, CH60-90, CH30-90 groups. Basal brain glucose consumption and hypothalamic insulin receptor density were lower only in the CH30-60 group. The rate of brain lipogenesis was increased in the H30-90 and CH30-90 groups.

Conclusion

These findings indicate that both H and CH diet regimens increased body adiposity independent treatment and the age at which treatment was started, whereas these diets caused hyperglycemia and affected brain metabolism when started at an early age.
On the solvability of a boundary value problem on the real line
Authors: Cupini Giovanni , Marcelli Cristina , Papalini Francesca
Published: September 2011

We investigate the existence of heteroclinic solutions to a class of nonlinear differential equations

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a

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)

Φ

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=
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)

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.
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governed by a nonlinear differential operator Φ extending the classical p- Laplacian, with right-hand side f having the critical rate of decay -1 as |t| → +∞, that is

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≈

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. We prove general existence and non-existence results, as well as some simple criteria useful for right-hand side having the product structure f(t, x, x') = b(t, x)c(x, x').

Mathematical subject classification: Primary: 34B40; 34C37; Secondary: 34B15; 34L30.
Chotosan (Diaoteng San)-induced improvement of cognitive deficits in senescence-accelerated mouse (SAMP8) involves the amelioration of angiogenic/neurotrophic factors and neuroplasticity systems in the brain
Authors: Zhao Qi , Yokozawa Takako , Tsuneyama Koichi , Tanaka Ken , Miyata Takeshi , Shibahara Notoshi , Matsumoto Kinzo
Published: September 2011

Background

Chotosan (CTS, Diaoteng San), a Kampo medicine (ie Chinese medicine) formula, is reportedly effective in the treatment of patients with cerebral ischemic insults. This study aims to evaluate the therapeutic potential of CTS in cognitive deficits and investigates the effects and molecular mechanism(s) of CTS on learning and memory deficits and emotional abnormality in an animal aging model, namely 20-week-old senescence-accelerated prone mice (SAMP8), with and without a transient ischemic insult (T2VO).

Methods

Age-matched senescence-resistant inbred strain mice (SAMR1) were used as control. SAMP8 received T2VO (T2VO-SAMP8) or sham operation (sham-SAMP8) at day 0. These SAMP8 groups were administered CTS (750 mg/kg, p.o.) or water daily for three weeks from day 3.

Results

Compared with the control group, both sham-SAMP8 and T2VO-SAMP8 groups exhibited cognitive deficits in the object discrimination and water maze tests and emotional abnormality in the elevated plus maze test. T2VO significantly exacerbated spatial cognitive deficits of SAMP8 elucidated by the water maze test. CTS administration ameliorated the cognitive deficits and emotional abnormality of sham- and T2VO-SAMP8 groups. Western blotting and immunohistochemical studies revealed a marked decrease in the levels of phosphorylated forms of neuroplasticity-related proteins, N-methyl-D-aspartate receptor 1 (NMDAR1), Ca2+/calmodulin-dependent protein kinase II (CaMKII), cyclic AMP responsive element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) in the frontal cortices of sham-SAMP8 and T2VO-SAMP8. Moreover, these animal groups showed significantly reduced levels of vasculogenesis/angiogenesis factors, vascular endothelial growth factor (VEGF), VEGF receptor type 2 (VEGFR2), platelet-derived growth factor-A (PDGF-A) and PDGF receptor α (PDGFRα). CTS treatment reversed the expression levels of these factors down-regulated in the brains of sham- and T2VO-SAMP8.

Conclusion

Recovery of impaired neuroplasticity system and VEGF/PDGF systems may play a role in the ameliorative effects of CTS on cognitive dysfunction caused by aging and ischemic insult.
The physiologic responses to epinephrine during cooling and after rewarming in vivo
Authors: Tveita Torkjel , Sieck Gary C
Published: September 2011

Introduction

The purpose of our study was to determine whether hypothermia has any effects on physiological hemodynamic responses to epinephrine (Epi), and whether rewarming reverses these effects.

Methods

Sprague-Dawley rats were instrumented to measure mean arterial pressure (MAP), and left ventricular (LV) pressure-volume changes were recorded by using a Millar pressure-volume conductance catheter. Core temperature was reduced from 37°C to 28°C and returned to 37°C by using both internal and external heat exchangers. Two groups of rats were infused with either saline (n = 7), or Epi 0.125 μg/min continuously (n = 7). At 33°C, 30°C, and 28°C, the Epi infusion was temporarily increased from 0.125 to 1.25 μg/min.

Results

Before cooling, Epi infusion in both groups resulted in a significant, dose-dependent increase in heart rate (HR), stroke volume (SV), cardiac output (CO), LV dP/dtmax (maximum derivative of systolic pressure over time), but only Epi infusion at 1.25 μg/min caused elevation of MAP. During cooling to 30°C, Epi infusion at 0.125 μg/min caused a significant elevation of central hemodynamic variables, whereas MAP remained unchanged. In contrast, Epi infusions at 1.25 μg/min caused a significant elevation of MAP during cooling to 28°C but no increases in central hemodynamics. After rewarming, all hemodynamic variables returned to baseline in both groups, but only the saline-treated animals displayed the prehypothermic hemodynamic dose responses to Epi infusions.

Conclusions

This study shows that hypothermia causes a change in the physiological hemodynamic response to Epi, which is not reversed by rewarming.
Cost utility of behavioural activation delivered by the non-specialist
Authors: Ekers David , Godfrey Christine , Gilbody Simon , Parrott Steve , Richards David A. , Hammond Danielle , Hayes Adele
Published: September 2011

Behavioural activation by non-specialists appears effective in the treatment of depression. We examined incremental cost-effectiveness of behavioural activation (n= 24)v.treatment as usual (n= 23) in a randomised controlled trial. Intention-to-treat analyses indicated a quality-adjusted life-year (QALY) difference in favour of behavioural activation of 0.20 (95% CI 0.01–0.39,P= 0.042), incremental cost-effectiveness ratio of £5756 per QALY and a 97% probability that behavioural activation is more cost-effective at a threshold value of £20 000. Results are promising for dissemination of behavioural activation but require replication in a larger study.
Dying well with dementia: Qualitative examination of end-of-life care
Authors: Lawrence Vanessa , Samsi Kritika , Murray Joanna , Harari Danielle , Banerjee Sube
Published: September 2011

Background

People with dementia often die badly, receiving end-of-life care of poorer quality than that given to those who are cognitively intact.

Aims

To define good end-of-life care for people with dementia and identify how it can be delivered across care settings in the UK.

Method

In-depth interviews were conducted with 27 bereaved family carers and 23 care professionals recruited from the community, care homes, general hospitals and continuing care units. Data were analysed using the constant comparison method.

Results

The data highlighted the challenge and imperative of ‘dementia-proofing’ end-of-life care for people with dementia. This requires using dementia expertise to meet physical care needs, going beyond task-focused care and prioritising planning and communication with families.

Conclusions

The quality of end-of-life care exists on a continuum across care settings. Together, the data reveal key elements of good end-of-life care and that staff education, supervision and specialist input can enable its provision.