bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression ofStMYB1R-1, a Novel Potato Single MYB-Like Domain Transcription Factor, Increases Drought Tolerance
Authors: Shin Dongjin , Moon Seok-Jun , Han Seyoun , Kim Beom-Gi , Park Sang Ryeol , Lee Seong-Kon , Yoon Hye-Jin , Lee Hye Eun , Kwon Hawk-Bin , Baek Dongwon , Yi Bu Young , Byun Myung-Ok
Published: October 2010

Potato (Solanum tuberosum) is relatively vulnerable to abiotic stress conditions such as drought, but the tolerance mechanisms for such stresses in potato are largely unknown. To identify stress-related factors in potato, we previously carried out a genetic screen of potato plants exposed to abiotic environmental stress conditions using reverse northern-blot analysis. A cDNA encoding a putative R1-type MYB-like transcription factor (StMYB1R-1) was identified as a putative stress-response gene. Here, the transcript levels of StMYB1R-1 were enhanced in response to several environmental stresses in addition to drought but were unaffected by biotic stresses. The results of intracellular targeting and quadruple 9-mer protein-binding microarray analysis indicated that StMYB1R-1 localizes to the nucleus and binds to the DNA sequence G/AGATAA. Overexpression of a StMYB1R-1 transgene in potato plants improved plant tolerance to drought stress while having no significant effects on other agricultural traits. Transgenic plants exhibited reduced rates of water loss and more rapid stomatal closing than wild-type plants under drought stress conditions. In addition, overexpression of StMYB1R-1 enhanced the expression of drought-regulated genes such as AtHB  -  7, RD28, ALDH22a1, and ERD1-like. Thus, the expression of StMYB1R-1 in potato enhanced drought tolerance via regulation of water loss. These results indicated that StMYB1R-1 functions as a transcription factor involved in the activation of drought-related genes.
SHORT-ROOT Regulates Primary, Lateral, and Adventitious Root Development in Arabidopsis
Authors: Lucas Mikaël , Swarup Ranjan , Paponov Ivan A. , Swarup Kamal , Casimiro Ilda , Lake David , Peret Benjamin , Zappala Susan , Mairhofer Stefan , Whitworth Morag , Wang Jiehua , Ljung Karin , Marchant Alan , Sandberg Goran , Holdsworth Michael J. , Palme Klaus , Pridmore Tony , Mooney Sacha , Bennett Malcolm J.
Published: October 2010

SHORT-ROOT (SHR) is a well-characterized regulator of radial patterning and indeterminacy of the Arabidopsis (Arabidopsis thaliana) primary root. However, its role during the elaboration of root system architecture remains unclear. We report that the indeterminate wild-type Arabidopsis root system was transformed into a determinate root system in the shr mutant when growing in soil or agar. The root growth behavior of the shr mutant results from its primary root apical meristem failing to initiate cell division following germination. The inability of shr to reactivate mitotic activity in the root apical meristem is associated with the progressive reduction in the abundance of auxin efflux carriers, PIN-FORMED1 (PIN1), PIN2, PIN3, PIN4, and PIN7. The loss of primary root growth in shr is compensated by the activation of anchor root primordia, whose tissues are radially patterned like the wild type. However, SHR function is not restricted to the primary root but is also required for the initiation and patterning of lateral root primordia. In addition, SHR is necessary to maintain the indeterminate growth of lateral and anchor roots. We conclude that SHR regulates a wide array of Arabidopsis root-related developmental processes.
Reverse-Genetic Analysis of the Two Biotin-Containing Subunit Genes of the Heteromeric Acetyl-Coenzyme A Carboxylase in Arabidopsis Indicates a Unidirectional Functional Redundancy
Authors: Li Xu , Ilarslan Hilal , Brachova Libuse , Qian Hui-Rong , Li Ling , Che Ping , Wurtele Eve Syrkin , Nikolau Basil J.
Published: October 2010

The heteromeric acetyl-coenzyme A carboxylase catalyzes the first and committed reaction of de novo fatty acid biosynthesis in plastids. This enzyme is composed of four subunits: biotin carboxyl-carrier protein (BCCP), biotin carboxylase, α-carboxyltransferase, and β-carboxyltransferase. With the exception of BCCP, single-copy genes encode these subunits in Arabidopsis (Arabidopsis thaliana). Reverse-genetic approaches were used to individually investigate the physiological significance of the two paralogous BCCP-coding genes, CAC1A (At5g16390, codes for BCCP1) and CAC1B (At5g15530, codes for BCCP2). Transfer DNA insertional alleles that completely eliminate the accumulation of BCCP2 have no perceptible effect on plant growth, development, and fatty acid accumulation. In contrast, transfer DNA insertional null allele of the CAC1A gene is embryo lethal and deleteriously affects pollen development and germination. During seed development the effect of the cac1a null allele first becomes apparent at 3-d after flowering, when the synchronous development of the endosperm and embryo is disrupted. Characterization of CAC1A antisense plants showed that reducing BCCP1 accumulation to 35% of wild-type levels, decreases fatty acid accumulation and severely affects normal vegetative plant growth. Detailed expression analysis by a suite of approaches including in situ RNA hybridization, promoter:reporter transgene expression, and quantitative western blotting reveal that the expression of CAC1B is limited to a subset of the CAC1A-expressing tissues, and CAC1B expression levels are only about one-fifth of CAC1A expression levels. Therefore, a likely explanation for the observed unidirectional redundancy between these two paralogous genes is that whereas the BCCP1 protein can compensate for the lack of BCCP2, the absence of BCCP1 cannot be tolerated as BCCP2 levels are not sufficient to support heteromeric acetyl-coenzyme A carboxylase activity at a level that is required for normal growth and development.
Role of AtPolζ, AtRev1, and AtPolη in UV Light-Induced Mutagenesis in Arabidopsis
Authors: Nakagawa Mayu , Takahashi Shinya , Tanaka Atsushi , Narumi Issay , Sakamoto Ayako N.
Published: October 2010

Translesion synthesis (TLS) is a DNA damage tolerance mechanism in which DNA lesions are bypassed by specific polymerases. To investigate the role of TLS activities in ultraviolet light-induced somatic mutations, we analyzed Arabidopsis (Arabidopsis thaliana) disruptants of AtREV3, AtREV1, and/or AtPOLH genes that encode TLS-type polymerases. The mutation frequency in rev3-1 or rev1-1 mutants decreased compared with that in the wild type, suggesting that AtPolζ and AtRev1 perform mutagenic bypass events, whereas the mutation frequency in the polh-1 mutant increased, suggesting that AtPolη performs nonmutagenic bypass events with respect to ultraviolet light-induced lesions. The rev3-1 rev1-1 double mutant showed almost the same mutation frequency as the rev1-1 single mutant. The increased mutation frequency found in polh-1 was completely suppressed in the rev3-1 polh-1 double mutant, indicating that AtPolζ is responsible for the increased mutations found in polh-1. In summary, these results suggest that AtPolζ and AtRev1 are involved in the same (error-prone) TLS pathway that is independent from the other (error-free) TLS pathway mediated by AtPolη.
Differentiation and adaptive radiation of amphibious gobies (Gobiidae: Oxudercinae) in semi‐terrestrial habitats
Authors: Polgar G. , Sacchetti A. , Galli P.
Published: October 2010

During several surveys made in the region of the lower Fly River and delta, Papua New Guinea, nine species of oxudercine gobies (Gobiidae: Oxudercinae) were recorded: Boleophthalmus caeruleomaculatus, Oxuderces wirzi, Periophthalmodon freycineti, Periophthalmus darwini, Periophthalmus novaeguineaensis, Periophthalmus takita, Periophthalmus weberi, Scartelaos histophorus and Zappa confluentus. An exploratory multivariate analysis of their habitat conditions discriminated five guilds, differentially distributed in habitats with different quantities of environmental water and three guilds corresponding to different levels of salinity. A partial correspondence between phylogenetic and ecological categories suggested the presence of parallel adaptive radiations within different genera. In particular, the species found in the most terrestrial habitats (P. weberi) was also found in the widest range of conditions, suggesting that colonization of extreme semi‐terrestrial and freshwater habitats by this species was facilitated by eurytypy. It is proposed that these findings provide insight into convergent adaptations for the vertebrate eco‐evolutionary transition from sea to land.
Telomerase activation without shortening of telomeric 3′‐overhang is a poor prognostic factor in human colorectal cancer
Authors: Kojima Kensaku , Hiyama Eiso , Otani Keiko , Ohtaki Megu , Fukuba Ikuko , Fukuda Emi , Sueda Taijiro , Hiyama Keiko
Published: October 2010

Our previous report demonstrated a good correlation between high telomerase activity of cancer tissues and a poor prognosis of patients with colorectal cancers, except for several cases. To elucidate the additional factors that contribute to patient prognosis, the correlation among the expression levels of telomere binding proteins (TBP), the lengths of telomeres, the lengths of telomere 3′‐overhang (3′‐OH) and telomerase activity in 106 paired colorectal cancer and corresponding noncancerous mucosa (NCM) specimens were examined. The expression levels of eight TBP genes (TRF1, TRF2, TIN2, TANK1, TANK2, POT1, RAP1 and TPP1) were analyzed. Among the 106 cases, 35 cases had shortened telomeres (<7 kb), 15 had shortened 3′‐OH (3′‐OH length ratio of cancer/NCM <0.5) and 88 were classified as telomerase‐activated cancers (activity ratio of cancer/NCM >2). Comparison between NCM and cancer in each case showed that all TBP except for POT1 were downregulated in cancers. A survival analysis using a Cox proportional hazard model showed that the survival rate of the telomerase‐activated cases with shortened 3′‐OH and that of telomerase‐inactivated cases were significantly better than that of telomerase‐activated cases without 3′‐OH shortening, that is, restored or maintained 3′‐OH (P = 0.018). In the telomerase‐activated cancers, the length of 3′‐OH was significantly correlated with the expression levels of POT1. Elongation of telomeric overhang by telomerase, which might be regulated by POT1, may contribute to the increase of malignant potential in colorectal cancers. (Cancer Sci 2011; 102: 330–335)
Modelling the adverse effects associated with ecstasy use
Authors: Fisk John E. , Murphy Philip N. , Montgomery Catharine , Hadjiefthyvoulou Florentia
Published: October 2010

Aims  Ecstasy, the street name for 3,4‐meththylenedioxymethamphetamine, has been associated with a range of psychiatric symptoms and impaired psychological health in both problem and recreational users. The purpose of the present paper is to determine how these impairments are related to the history of polydrug use, and the conditions under which individuals ingest ecstasy.

Design  Associations between the variables of interest were investigated utilizing negative binomial regression.

Setting  Liverpool and Preston in the North West of England.

Participants  A convenience sample of 159 recreational ecstasy/polydrug users (80 males, 79 females). The sample was composed primarily of undergraduates.

Measurements  The dependent variable was the number of reported ecstasy‐related adverse effects. Independent variables included quantitative aspects of ecstasy and other drug use, and the various beliefs and behaviours associated with ecstasy use.

Findings  The number of adverse effects was associated positively with life‐time exposure to ecstasy and negatively with period of abstinence from the drug. Adverse effects were more common among those who consumed ecstasy and alcohol concurrently, but were unrelated to other aspects of polydrug use. They were unaffected by whether the user took precautions when using the drug, and only weakly related to prior beliefs concerning the effects of ecstasy.

Conclusions  Greater life‐time exposure to ecstasy and consuming the drug concurrently with alcohol increase the likelihood of experiencing adverse effects, including paranoia, poor general health, irritability, confusion and moodiness. Adverse effects decrease with the period of abstinence from the drug.
Review article: medical, surgical and radiological management of perianal Crohn’s fistulas
Authors: Tozer P. J. , Burling D. , Gupta A. , Phillips R. K. S. , Hart A. L.
Published: October 2010

Aliment Pharmacol Ther 2011; 33: 5–22

Background  Crohn’s anal fistulas are common and cause considerable morbidity. Their management is often difficult; medical and surgical treatments rarely lead to true healing with frequent recurrence and complications.

Aim  To examine medical treatments previously and currently used, surgical techniques and the important role of optimal imaging.

Methods  We conducted a literature search in the Pub Med database using Crohn’s, Anal Fistula, Surgery, Imaging and Medical Treatment as search terms.

Results  Antibiotics and immunosuppressants have a role, but slow initial response, side effects and relatively low remission rates of up to around a third with frequent recurrence limit their value. Long‐term infliximab produces clinical remission in 36–58% of patients with combined medical and surgical management achieving optimal outcomes. Traditional and newer surgical procedures often have a high rate of recurrence with a significant risk of temporary or, in up to 10% of cases, permanent stomas, incontinence and unhealed or slowly healing wounds in 30%.

Conclusions  Management of Crohn’s anal fistulas remains challenging. Established principles are to drain infection, use setons as required, aggressively manage active proctitis, give antibiotics, immunosuppressants and employ anti‐TNFα therapy, and they demand significant co‐operation between gastroenterologists and surgeons.
Increased proportion of CD16+ NK cells in the colonic lamina propria of inflammatory bowel disease patients, but not after azathioprine treatment
Authors: Steel A. W. , Mela C. M. , Lindsay J. O. , Gazzard B. G. , Goodier M. R.
Published: October 2010

Aliment Pharmacol Ther 2011; 33: 115–126

Background  Distinct functional subsets of natural killer cells potentially contribute to the pathology of inflammatory bowel disease (IBD).

Aim  To report the phenotypic and functional characteristics of natural killer cells in blood and lamina propria of IBD patients, and the effect of azathioprine.

Methods  Natural killer cells from blood and lamina propria of healthy controls or patients with Crohn’s disease, or ulcerative colitis were studied by flow cytometry. Activation, cytokine production, proliferation and apoptosis of natural killer cell subsets were studied in vitro.

Results  CD16+ natural killer cells are increased in frequency in the lamina propria comparing Crohn’s disease or ulcerative colitis with healthy controls. Azathioprine therapy was associated with a reduction in total natural killer cells in blood and lamina propria, preferentially of the CD16+ subset. Azathioprine therapy did not impair natural killer degranulation, but reduced natural and cytokine‐activated cytotoxicity and interferon‐gamma (IFN‐γ) production. Culture of resting peripheral blood mononuclear cells with azathioprine resulted in loss of natural killer cells and inhibition of activation and IFN‐γ production. Azathioprine preferentially inhibited proliferation of CD16+ natural killer cells and induced apoptosis in resting but not in pre‐activated natural killer cells.

Conclusions  Natural killer cells with cytolytic potential are enriched in the colonic lamina propria of individuals with IBD. Azathioprine is associated with a reduction in these cells and a normalization of natural killer cell populations.
Randomised clinical trial: identification of responders to short‐term treatment with esomeprazole for dyspepsia in primary care – a randomised, placebo‐controlled study
Authors: Meineche‐Schmidt V. , Christensen E. , Bytzer P.
Published: October 2010

Aliment Pharmacol Ther 2011; 33: 41–49

Background  Response to proton pump inhibitor (PPI) treatment in dyspepsia is unpredictable.

Aim  To identify symptoms associated with response to esomeprazole in order to target patients for empirical treatment.

Methods  Eight hundred and five uninvestigated, primary care patients with upper GI symptoms that were considered to be acid‐related were randomised to 2 weeks’ treatment with esomeprazole 40 mg or placebo. The study population was divided into a model sample (N = 484) and a validation sample (N = 321). We developed a therapeutic index to predict PPI response from the model sample and tested this in the validation sample.

Results  Response to PPI was found in 68% of patients (44% in placebo arm). Bothersome heartburn and early satiety were associated with increased likelihood of PPI response, whereas dull abdominal pain, pain relieved by bowel movements and nausea in women were associated with a decreased likelihood of PPI response. Patients in the validation sample could be classified as having a ‘very high’ (n = 55), ‘high’ (n = 123), ‘medium’ (n = 78) or ‘low’ (n = 65) probability of PPI response. The therapeutic gains over placebo were 55%, 31%, 20% and 22%, respectively.

Conclusions  In patients with uninvestigated dyspepsia, PPI responders can be reliably identified by a simple pocket chart using symptoms and patient characteristics (ClinicalTrials.gov NCT00318968).
Trends in co‐prescribing of angiotensin converting enzyme inhibitors and angiotensin receptor blockers in Ireland
Authors: Wan Md Adnan Wan A. H. , Zaharan Nur L. , Bennett Kathleen , Wall Catherine A.
Published: October 2010

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Renin‐angiotensin‐aldosterone system inhibition confers cardio‐renal protection and may be achieved by monotherapy with an angiotensin enzyme converting inhibitor (ACEI) or an angiotensin‐II receptor blocker (ARB).

• Dual ACEI and ARB therapy has been examined in major clinical trials with conflicting results.

• The prescribing pattern of dual therapy in general primary care population is not known.

WHAT THIS STUDY ADDS

• An increase in the co‐prescribing of ACEIs and ARBs was observed in the Irish primary care population.

• Co‐prescribing of ACEIs and ARBs in the primary care population did not appear to be influenced by results from major clinical trials.

• ACEIs and ARBs were more likely to be co‐prescribed in special cohorts of the population with diabetes, hypertension and heart failure.

AIMS

(i) To examine the trends in co‐prescribing of angiotensin converting enzyme inhibitor (ACEI) and angiotensin‐II receptor blocker (ARB) therapy and (ii) to examine the influence of major clinical trials (CALM, COOPERATE, VALIANT and ONTARGET) on co‐prescribing.

METHODS

The Irish HSE‐Primary Care Reimbursement Services database was used to identify patients ≥16 years old co‐prescribed ACEIs and ARBs between January 2000 and April 2009 (n= 266 554 prescriptions). The rate of prescribing per 1000 general medical services (GMS) scheme population was calculated for each month. Patients with diabetes, hypertension, heart failure and ischaemic heart disease were also identified by prescribing of certain medications. A linear trend test was used to examine prescribing trends. Logistic regression was used to examine prescribing according to patient characteristics. The effects of the major trials on prescribing were examined using segmented regression analysis for 12 months pre‐ and post‐trials.

RESULTS

There was a significant linear trend in overall ACEI and ARB co‐prescribing over the study period (P < 0.001). Rate of co‐prescribing in January 2000 and April 2009 was 0.16 and 5.72, per 1000 eligible population, respectively. Those 45–64 years old (OR = 2.88, 95% confidence interval (CI) 2.71, 3.06) and ≥65 years (OR = 2.52, 95% CI 2.36, 2.68) were more likely to receive dual therapy compared with those <45 years old. Those with hypertension (OR = 8.85, 95% CI 8.45, 9.27), diabetes (OR = 4.10, 95% CI 3.97, 4.23) and heart failure (OR = 1.78, 95% CI 1.72, 1.84) were more likely to receive dual therapy compared with the general population. Significant increases in prescribing were observed only after the CALM (P= 0.03) and VALIANT (P= 0.007) trials.

CONCLUSION

Increased co‐prescribing of ACEIs and ARBs was observed in Ireland during 2000–09. Prescribing patterns did not appear to be affected by results from major trials.
Improving the quality of drug research or simply increasing its cost? An evidence‐based study of the cost for data monitoring in clinical trials
Authors: Pronker Esther , Geerts Bart F. , Cohen Adam , Pieterse Herman
Published: October 2010

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Current knowledge is based on practical experience. This paper presents the first novel, quantitative and qualitative view on the efficacy of the sponsor query system widely applied in the pharmaceutical industry, used for quality control purposes by data management for clinical data.

WHAT THIS STUDY ADDS

• This study presents a structured view on the process of clinical data management and sponsor queries, the efficacy and cost effectiveness of sponsor queries and dual data entry and the conclusion to advocate a more evidence‐based approach to clinical data validation.

AIM

Procedures for verification of data from clinical studies are intended to maintain reliability for clinical trial results. Guidelines or legislations relating to clinical data management are of limited value and no study has yet demonstrated its effectiveness.

METHOD

Sponsor queries and dual entry procedures from one CRO on three different phase I trials are analysed on content, impact and cost.

RESULT

In this study, sponsor queries and dual entry procedures proved time and cost inefficient in detecting data discrepancies.

CONCLUSION

We advocate a more evidence‐based approach for enhancing data integrity throughout the process of clinical data management.
A point prevalence survey of antibiotic prescriptions: benchmarking and patterns of use
Authors: Aldeyab Mamoon A. , Kearney Mary P. , McElnay James C. , Magee Fidelma A. , Conlon Geraldine , Gill Dianne , Davey Peter , Muller Arno , Goossens Herman , Scott Michael G. ,   on behalf of the ESAC Hospital Care Subproject Group
Published: October 2010

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Inappropriate antimicrobial use has been associated with increased morbidity and hospital costs.

• Antibiotic policies aim to improve patient outcomes whilst reducing adverse effects associated with antimicrobial use.

• More insight into the actual implementation of antibiotic policies is needed in order to explore patterns of antibiotic prescribing.

WHAT THIS STUDY ADDS

• This study assessed the current patterns of antibiotic prescribing and the impact of a hospital antibiotic policy on these practices.

• It demonstrated the value of point prevalence surveys in informing antibiotic stewardship and identifying targets for quality improvements.

• The study emphasized the importance of participating in international networks, such as the European Surveillance of Antimicrobial Consumption (ESAC), in supporting optimal antibiotic use.

AIM The aim of the study was to assess current patterns of antibiotic prescribing and the impact of a hospital antibiotic policy on these practices.

METHODS The study involved collecting information regarding hospitalized patients utilizing the ESAC audit tool.

RESULTS In the study site hospital, the use of the restricted agents was low whilst the use of the non‐restricted agents was high. Compliance with the hospital antibiotic guidelines was 70%.

DISCUSSION The findings identified monitoring non‐restricted antibiotics and compliance with guidelines as targets for quality improvements in our hospital. Point prevalence surveys may offer a simple method of monitoring antibiotic policies, thus, informing antibiotic stewardship.
Ion channels in volume regulation of clonal kidney cells
Authors: Da Silva M. B. , Costa V. M. A. , Pereira V. R. A. , De Albertim G. J. B. , De Melo E. B. B. , Bezerra D. P. , Da Silva R. P. , Rodrigues C. G. , Carneiro C. M. M. , Yuldasheva L. N. , Krasilnikov O. V.
Published: October 2010

Objectives:  Clonal kidney cells (Vero cells) are extensively utilized in the manufacture of biological preparations for disease diagnostics and therapeutics and also in preparation of vaccines. In all cells, regulation of volume is an essential function coupled to a variety of physiological processes and is a topic of interest. The objective here was to investigate involvement of ion channels in the process of volume regulation of Vero cells.

Methods:  Involvement of ion channels in cell volume regulation was studied using video‐microscopy and flow cytometry. Pharmacologically unaltered cells of different sizes, which are presumably at different phases of the cell cycle, were used.

Results:  Ion transport inhibitors altered all phases of regulatory volume decrease (RVD) of Vero cells, rate of initial cell swelling, Vmax and volume recovery. Effects were dependent on type of inhibitor and on cell size (cell cycle phase). Participation of aquaporins in RVD was suggested. Inhibitors decelerated growth, arresting Vero cells at the G0/G1 phase boundary. Electrophysiological study confirmed presence of volume‐activated Cl− channels and K+ channels in plasmatic membranes of the cells.

Conclusion:  Vero cells of all sizes maintained the ability to recover from osmotic swelling. Activity of ion channels was one of the key factors that controlled volume regulation and proliferation of the cells.
Coupled mathematical model of tumorigenesis and angiogenesis in vascular tumours
Authors: Cooper M. D. , Tanaka M. L. , Puri I. K.
Published: October 2010

Objectives: Mathematical models are useful for studying vascular and avascular tumours, because these allow for more logical experimental design and provide valuable insights into the underlying mechanisms of their growth and development. The processes of avascular tumour growth and the development of capillary networks through tumour‐induced angiogenesis have already been extensively investigated, albeit separately. Despite the clinical significance of vascular tumours, few studies have combined these approaches to develop a single comprehensive growth and development model.

Materials and methods: We develop a continuum‐based mathematical model of vascular tumour growth. In the model, angiogenesis is initiated through the release of angiogenic growth factors (AGFs) by cells in the hypoxic regions of the tumour. The nutrient concentration within the tumour reflects the influence of capillary growth and invasion induced by AGF.

Results and conclusions: Parametric and sensitivity studies were performed to evaluate the influence of different model parameters on tumour growth and to identify the parameters with the most influence, which include the rates of proliferation, apoptosis and necrosis, as well as the diffusion of sprout tips and the size of the region affected by angiogenesis. An optimization was performed for values of the model parameters that resulted in the best agreement with published experimental data. The resulting model solution matched the experimental data with a high degree of correlation (r = 0.85).
Improved procedure for the measurement of telomere length in whole cells by PNA probe and flow cytometry
Authors: Carbonari M. , Mancaniello D. , Cibati M. , Catizone A. , Fiorilli M.
Published: October 2010

Objectives:  Peptide nucleic acid (PNA) probes hybridize to denatured telomeric sequences in cells permeabilized in hot formamide. In reported protocols, the hybridization was conducted in solutions with high formamide concentrations to avoid the DNA renaturation that can hamper binding of the oligo‐PNA probe to specific sequences. We postulated that telomeric DNA, confined in the nuclear microvolume, is not able to properly renature after hot formamide denaturation. Therefore, to improve hybridization conditions between the probe and the target sequences, it might be possible to add probe to sample after the complete removal of formamide.

Materials and methods:  After telomeric DNA denaturation in hot formamide solution and several washes to remove the ionic solvent, cells were hybridized overnight at room temperature with human telomere‐specific PNA probe conjugated with Cy5 fluorochrome, Cy5‐OO‐(CCCTAA)3. After stringency washes and staining with ethidium bromide, the cells were analysed by flow cytometry and by using a confocal microscope.

Results:  Using three continuous cell lines, different in DNA content and telomere length, and resting human peripheral blood T and B lymphocytes, we demonstrated that the oligo‐PNA probe hybridized to telomeric sequences after complete removal of formamide and that in the preserved nucleus, telomeric sequence denaturation is irreversible.

Conclusion:  According to our experience, oligo‐PNA binding results is efficient, specific and proportional to telomere length. These, our original findings, can form the technological basis of actual in situ hybridization on preserved whole cells.
Assessment of cell proliferation and muscular structure following surgical tongue volume reduction in pigs
Authors: Ye W. , Abu A. F. , Liu Z. J.
Published: October 2010

Objectives:  Tongue volume reduction is an adjunct treatment in several orofacial orthopaedic procedures for various craniofacial deformities; it may affect structural reconstitution and functional recovery as a result of the repair process. The aim of this study was to investigate myogenic regeneration and structural alteration of the tongue following surgical tongue volume reduction.

Materials and methods:  Five 12‐week‐old sibling pairs of Yucatan minipigs (three males and two females) were used. Midline uniform glossectomy was performed on one of each pair (reduction); siblings had identical incisions without tissue removal (sham). All pigs were raised for a further 4 weeks and received 5‐bromo‐2‐deoxyuridine (BrdU) injection intravenously 1 day before killing. Tissue sections of tongues were stained with anti‐BrdU antibody to evaluate numbers of replicating cells. Haematoxylin and eosin plus trichrome staining were performed to assess muscular structure.

Results:  Reduction tongues contained significantly more BrdU+ cells compared to sham tongues (P < 0.01). However, these BrdU+ cells were mostly identified in reparative connective tissues (fibroblasts) rather than in regenerating muscle tissue (myoblasts). Trichrome‐stained sections showed disorganized collagen fibres linked to few intermittent muscle fibres in the reduction tongues. These myofibres presented signs of atrophy with reduced perimysium and endomysium. Matrix between reduced perimysium and endomysium was filled with fibrous tissue.

Conclusions:  Fibrosis without predominant myogenic regeneration was the major histological consequence of surgical tongue volume reduction.
Identification of tyrosine‐nitrated proteins in HT22 hippocampal cells during glutamate‐induced oxidative stress
Authors: Yoon S.‐W. , Kang S. , Ryu S.‐E. , Poo H.
Published: October 2010

Objectives:  Nitration of tyrosine residues in protein is a post‐translational modification, which occurs under oxidative stress, and is associated with several neurodegenerative diseases. To understand the role of nitrated proteins in oxidative stress‐induced cell death, we identified nitrated proteins and checked correlation of their nitration in glutamate‐induced HT22 cell death.

Materials and methods:  Nitrated proteins were detected by western blotting using an anti‐nitrotyrosine antibody, extracted from matching reference 2‐dimensional electrophoresis gels, and identified with matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry.

Results:  Glutamate treatment induced apoptosis in HT22 cells, while reactive oxygen species (ROS) inhibitor or neuronal nitric oxide synthase (nNOS) inhibitor blocked glutamate‐induced HT22 cell death. Nitration levels of 13 proteins were increased after glutamate stimulation; six of them were involved in regulation of energy production and two were related to apoptosis. The other nitrated proteins were associated with calcium signal modulation, ER dysfunction, or were of unknown function.

Conclusions:  The 13 tyrosine‐nitrated proteins were detected in these glutamate‐treated HT22 cells. Results demonstrated that cell death, ROS accumulation and nNOS expression were related to nitration of protein tyrosine in the glutamate‐stimulated cells.
Genistein and daidzein repress adipogenic differentiation of human adipose tissue‐derived mesenchymal stem cells via Wnt/β‐catenin signalling or lipolysis
Authors: Kim M.‐H. , Park J.‐S. , Seo M.‐S. , Jung J.‐W. , Lee Y.‐S. , Kang K.‐S.
Published: October 2010

Objectives:  One aspect of the effects of isoflavones against fat deposition might be at least associated with the mechanism by which Wnt/β‐catenin signalling inhibits adipocyte differentiation. However, it remains completely unknown as to whether isoflavones might influence Wnt signalling during commitment of pluripotent mesenchymal stem cells (MSCs) to adipose lineages. In the present study, we have investigated the mechanisms underlying effects of genistein and daidzein, the major soy isoflavones, on anti‐adipogenic Wnt/β‐catenin signalling.

Materials and methods:  Adipose tissue‐derived (AD) MSCs were exposed continuously to genistein and daidzein (0.01–100 μm) during adipogenic differentiation (21 days). An oestrogen antagonist, ICI 182,780, was used to determine whether or not the isoflavones activated Wnt signalling via oestrogen receptors (ERs).

Results:  Genistein and daidzein suppressed adipogenic differentiation of AD‐MSCs in a dose‐dependent manner and inhibited expression of adipogenic markers, PPARγ, SREBP‐1c and Glut 4, from mid‐phase differentiation. Microarrays showed that anti‐adipogenic effects of genistein were principally attributable to activation of Wnt signalling via ERs‐dependent pathway, such as Erk/JNK signalling and LEF/TCF4 co‐activators. These findings were supported by evidence that the effects of genistein were offset by ICI182,780. Unlike genistein, daidzein inhibited adipogenesis through stimulation of lipolysis, with for example, PKA‐mediated hormone sensitive lipase. This is consistent with the increase in glycerol released from AD‐MSCs. In conclusion, understanding that different sets of mechanisms of the two isoflavones on adipogenesis will help the design of novel strategies to prevent observed current epidemic levels of obesity, using isoflavones.
Schwann‐like cell differentiation of rat adipose‐derived stem cells by indirect co‐culture with Schwann cells in vitro
Authors: Wei Y. , Gong K. , Zheng Z. , Liu L. , Wang A. , Zhang L. , Ao Q. , Gong Y. , Zhang X.
Published: October 2010

Objectives:  Schwann cell (SC) transplantation is a promising therapy for peripheral nerve transaction, however, clinical use of SCs is limited due to their very limited availability. Adipose‐derived stem cells (ADSCs) have been identified as an alternative source of adult stem cells in recent years. The aim of this study was to evaluate the feasibility of using ADSCs as a source of stem cells for differentiation into Schwann‐like cells by an indirect co‐culture approach, in vitro.

Materials and methods:  Multilineage differentiation potential of the obtained ADSCs was assayed by testing their ability to differentiate into osteoblasts and adipocytes. The ADSCs were co‐cultured with SCs to be induced into Schwann‐like cells through proximity, using a Millicell system. Expression of typical SC markers S‐100, GFAP and P75NTR of the treated ADSCs was determined by immunocytochemical staining, western blotting and RT‐PCR. Myelination capacity of the differentiated ADSCs (dADSCs) was evaluated in dADSC/dorsal root ganglia neuron (DRGN) co‐cultures.

Results:  The treated ADSCs adopted a spindle shaped‐like morphology after co‐cultured with SCs for 6 days. All results of immunocytochemical staining, western blotting and RT‐PCR showed that the treated cells expressed S‐100, GFAP and P75NTR, indications of differentiation. dADSCs could form Schwann‐like cell myelin in co‐culture with DRGNs. Undifferentiated ADSCs (uADSCs) did not form myelin compared to DRGNs cultured alone, but could produce neurite extension.

Conclusions:  These results demonstrate that this indirect co‐culture microenvironment could induce ADSCs to differentiate into Schwann‐like cells in vitro, which may be beneficial for treatment of peripheral nerve injuries in the near future.
Chronic colitis induces expression of β-defensins in murine intestinal epithelial cells
Authors: Rahman A , Fahlgren A , Sundstedt C , Hammarström S , Danielsson Å , Hammarström M-L
Published: October 2010

Anti-microbial peptides are important effectors in innate immunity. In the gut they defend against pathogens, shape the commensal microbiota and probably control intestinal homeostasis. Ulcerative colitis (UC), but not Crohn's disease, shows increased expression of inducible β-defensins (hBD-2, hBD-3 and hBD-4) in colonic epithelial cells. Does inducible defensin production precede the chronic intestinal inflammation characteristic of UC, or is it a consequence of the T cell-driven chronic inflammation? The aim was to analyse defensin mRNA and protein expression in colonic epithelial cells in two colitis mouse models resembling UC, the interleukin (IL)-2−/− mouse and the dextran sulphate sodium (DSS)-induced colitis mouse. Defensin mRNA was assayed by in situ hybridization and quantitative real-time reverse transcription–polymerase chain reaction (RT–PCR). Defensin peptide was assayed by immunohistochemistry. Mouse β-defensin 3 (mBD-3, orthologue to hBD-2) was up-regulated strongly in colonic epithelium of 15-week-old IL-2−/− mice and DSS-induced colitis mice with chronic bowel inflammation, but not in apparently healthy IL-2−/− 5-week-old mice, IL-2+/− 15-week-old mice or in acute stage DSS mice. Up-regulation was seen both at the mRNA- and at the protein level (only mBD-3 investigated). IL-17, but not several other cytokines, including interferon (IFN)-γ, induced mBD-3 mRNA expression in mouse colon carcinoma cells. The mRNA expression level of the constitutively expressed α-defensin, cryptdin-4, was up-regulated marginally in acute stage DSS-colitis mice and in IL-2−/− mice before signs of colitis. Inducible β-defensin expression in colonic epithelium is the consequence of the chronic bowel inflammation caused by activated T cells releasing cytokines including IL-17.
Predictors of Chinook salmon extirpation in California’s Central Valley
Authors: ZEUG S. C. , ALBERTSON L. K. , LENIHAN H. , HARDY J. , CARDINALE B.
Published: October 2010

Abstract  Chinook salmon, Oncorhynchus tshawytscha (Walbaum), populations have declined rapidly along the western coast of North America since the year 2000, possibly because of factors such as habitat loss, altered hydrology and barriers to migration. However, few analyses have rigorously examined which of these factors actually explain historical patterns of extirpation. Data were compiled on flow regimes, habitat loss and migration barriers for 27 streams that historically supported autumn run salmon and 22 streams that supported spring runs. The probability of extirpation in streams supporting autumn run was predicted solely by migration barriers. All other factors were >105 times less likely to explain existing variation. By contrast, models for spring run salmon suggest that habitat loss and altered flow regimes were also predictors of extirpation. These results suggest that regional extirpation of Chinook salmon has been driven by multiple forms of environmental change, and restoration efforts must address a multitude of bottlenecks that now impact spring and autumn run populations.
Genetic variation and community change – selection, evolution, and feedbacks
Authors: Genung Mark A. , Schweitzer Jennifer A. , Úbeda Francisco , Fitzpatrick Benjamin M. , Pregitzer Clara C. , Felker‐Quinn Emmi , Bailey Joseph K.
Published: October 2010

1. There is relatively little information on how evolutionary processes that alter genetic variation in a focal species may affect interactions with other species, impact the structure or function of the community and ecosystem, and affect evolutionary feedbacks among interacting species through time.

2. Because evolution can occur at ecological time‐scales, it is important to understand how major selective events, such as climatic changes, can impact the community of interacting species and ecosystem processes by changing intraspecific genetic variation.

3. The evidence linking genetic variation and evolution to community change and feedbacks has arisen from several different approaches whose results have not been synthesized into one conceptual framework, and whose commonalities may not be fully understood.

4. This review synthesizes several different experimental approaches on how evolution may impact communities and ecosystems and focuses on five main issues: (i) the genetic basis to communities and ecosystems; (ii) the community and ecosystem consequences of among‐population genetic differentiation; (iii) the role of local adaptation and co‐evolution; (iv) the effects of trans‐generational feedbacks and the eco‐evo dynamic and; (v) the integration of community and ecosystem genetics and multi‐level selection.

5. Evolution can alter intraspecific genetic variation to affect indirect genetic effects and feedbacks. Future studies should investigate how communities and ecosystems are affected when evolution causes the strength of feedbacks to change.
Role of SIRPα in regulation of mucosal immunity in the intestine
Authors: Kanazawa Yoshitake , Saito Yasuyuki , Supriatna Yana , Tezuka Hiroyuki , Kotani Takenori , Murata Yoji , Okazawa Hideki , Ohnishi Hiroshi , Kinouchi Yoshitaka , Nojima Yoshihisa , Ohteki Toshiaki , Shimosegawa Tooru , Matozaki Takashi
Published: October 2010

Mononuclear phagocytes such as dendritic cells (DCs) and macrophages in the lamina propria (LP) are thought to be important for both induction of inflammatory responses and maintenance of immunologic tolerance in the mammalian intestine. The molecular mechanisms by which these cells regulate intestinal immunity have remained poorly understood, however. Signal regulatory protein α (SIRPα) is a transmembrane protein that is specifically expressed in DCs, macrophages and neutrophils. Here, we show that SIRPα is abundant in CD11c+ CD11b+ LP cells of the mouse intestine. Whereas SIRPα did not appear to be important for the steady‐state homeostasis of mucosal immunity in the intestine, the flagellin‐stimulated production of IL‐17 or interferon (IFN)‐γ by LP cells of SIRPα mutant (MT) mice that lack the cytoplasmic region of the protein was markedly decreased compared with that observed with wild‐type cells. Moreover, the flagellin‐induced production of IL‐6 by LP cells from SIRPα MT mice was also greatly reduced. SIRPα MT mice were also resistant to the development of colitis induced by IL‐10 deficiency. Our data thus suggest that SIRPα expressed on CD11c+ LP cells is important for the production of IL‐17 or IFN‐γ in the LP as well as for the development of colitis induced by IL‐10 deficiency.
Diversity in immunological synapse structure
Authors: Thauland Timothy J. , Parker David C.
Published: October 2010

Immunological synapses (ISs) are formed at the T cell–antigen‐presenting cell (APC) interface during antigen recognition, and play a central role in T‐cell activation and in the delivery of effector functions. ISs were originally described as a peripheral ring of adhesion molecules surrounding a central accumulation of T‐cell receptor (TCR)–peptide major histocompatibility complex (pMHC) interactions. Although the structure of these ‘classical’ ISs has been the subject of intense study, non‐classical ISs have also been observed under a variety of conditions. Multifocal ISs, characterized by adhesion molecules dispersed among numerous small accumulations of TCR–pMHC, and motile ‘immunological kinapses’ have both been described. In this review, we discuss the conditions under which non‐classical ISs are formed. Specifically, we explore the profound effect that the phenotypes of both T cells and APCs have on IS structure. We also comment on the role that IS structure may play in T‐cell function.
Evaluating the impact of a new pay system on nurses in the UK
Authors: Buchan James , Ball Jane
Published: October 2010

Aims and objectives.  This study examines the impact of implementing a new pay system (Agenda for Change) on nursing staff in the National Health Service (NHS) in the UK. This new pay system covered approximately 400,000 nursing staff. Its objectives were to improve the delivery of patient care as well as staff recruitment, retention and motivation.

Background.  The new system aimed to provide a simplified approach to pay determination, with a more systematic use of agreed job descriptions and job evaluation to ‘price’ individual jobs, linked to a new career development framework.

Design.  Secondary analysis of survey data.

Methods.  Analysis of results of large‐scale surveys of members of the Royal College of Nursing of the United Kingdom (RCN) to assess the response of nurses to questions about the implementation process itself and their attitude to pay levels.

Results.  The results demonstrated that there was some positive change after implementation of Agenda for Change in 2006, mainly some time after implementation, and that the process of implementation itself raised expectations that were not fully met for all nurses.

Conclusions.  There were clear indications of differential impact and reported experiences, with some categories of nurse being less satisfied with the process of implementation. The overall message is that a national pay system has strengths and weaknesses compared to the local systems used in other countries and that these benefits can only be maximised by effective communication, adequate funding and consistent management of the system.

Relevance to clinical practice.  How nurses’ pay is determined and delivered can be a major satisfier and incentive to nurses if the process is well managed and can be a factor in supporting clinical practice, performance and innovation. This study highlights that a large‐scale national exercise to reform the pay system for nurses is a major undertaking, carries risk and will take significant time to implement effectively.
Relationships among sexual self‐concept, sexual risk cognition and sexual communication in adolescents: a structural equation model
Authors: Lou Jiunn‐Horng , Chen Sheng‐Hwang , Li Ren‐Hau , Yu Hsing‐Yi
Published: October 2010

Aim.  The purpose of this study was to test a model of sexual self‐concept and sexual risk cognition affecting sexual communication in Taiwanese adolescents.

Background.  Parent–adolescent sexual communication has been shown to influence adolescent sexual behaviour. Self‐concept is an important predictor of human behaviour, especially sexual behaviour. Few researchers have assessed sexual self‐concept in adolescents, despite its clear relevance to understanding adolescent sexual behaviour.

Design.  A cross‐sectional survey with convenience sampling was used in this study.

Method.  In 2009, data were collected by questionnaire from 748 adolescent students at a junior college in Taiwan.

Results.  The results revealed that the postulated model fits the data from this study well. Sexual self‐concept significantly predicts sexual risk cognition and sexual communication. Sexual risk cognition significantly predicts sexual communication and has an intervening effect on the relationship between sexual self‐concept and sexual communication.

Conclusions.  Sexual risk cognition is important in explaining sexuality in adolescents. Sexual self‐concept has both direct and indirect effects on sexual communication. Our findings provide concrete directions for school educators in developing sexual health programmes to increase adolescent sexual self‐concept and sexual communication with their parents.

Relevance to clinical practice.  Future sexual health programmes about sexual self‐concept and sexual risk cognition must add for increasing adolescent’s sexual communication with their parents.