bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evidence for different mechanisms of ‘unhooking’ for melphalan and cisplatin-induced DNA interstrand cross-links in vitroand in clinical acquired resistant tumour samples
Authors: Spanswick Victoria J , Lowe Helen L , Newton Claire , Bingham John P , Bagnobianchi Alessia , Kiakos Konstantinos , Craddock Charles , Ledermann Jonathan A , Hochhauser Daniel , Hartley John A 
Published: October 2012

Abstract

Background
DNA interstrand cross-links (ICLs) are critical lesions produced by several cancer chemotherapy agents including platinum drugs and nitrogen mustards. We have previously shown in haematological (multiple myeloma) and solid tumours (ovarian cancer) that clinical sensitivity to such agents can result from a defect in DNA ICL processing leading to their persistence. Conversely, enhanced repair can result in clinical acquired resistance following chemotherapy. The repair of ICLs is complex but it is assumed that the ‘unhooking’ step is common to all ICLs.

Methods
Using a modification of the single cell gel electrophoresis (Comet) assay we measured the formation and unhooking of melphalan and cisplatin-induced ICLs in cell lines and clinical samples. DNA damage response in the form of γ-H2AX foci formation and the formation of RAD51 foci as a marker of homologous recombination were also determined. Real-time PCR of 84 genes involved in DNA damage signalling pathways was also examined pre- and post-treatment.

Results
Plasma cells from multiple myeloma patients known to be clinically resistant to melphalan showed significant unhooking of melphalan-induced ICLs at 48 hours, but did not unhook cisplatin-induced ICLs. In ovarian cancer cells obtained from patients following platinum-based chemotherapy, unhooking of cisplatin-induced ICLs was observed at 48 hours, but no unhooking of melphalan-induced ICLs. In vitro, A549 cells were proficient at unhooking both melphalan and cisplatin-induced ICLs. γ-H2AX foci formation closely followed the formation of ICLs for both drugs, and rapidly declined following the peak of formation. RPMI8226 cells unhooked melphalan, but not cisplatin-induced ICLs. In these cells, although cross-links form with cisplatin, the γ-H2AX response is weak. In A549 cells, addition of 3nM gemcitabine resulted in complete inhibition of cisplatin-induced ICL unhooking but no effect on repair of melphalan ICLs. The RAD51 foci response was both drug and cell line specific. Real time PCR studies highlighted differences in the damage response to melphalan and cisplatin following equi-ICL forming doses.

Conclusions
These data suggest that the mechanisms by which melphalan and cisplatin-induced ICLs are ‘unhooked’ in vitro are distinct, and the mechanisms of clinical acquired resistance involving repair of ICLs, are drug specific.
A cluster-randomized controlled trial of strategies to increase adolescents’ physical activity and motivation during physical education lessons: the Motivating Active Learning in Physical Education (MALP) trial
Authors: Rosenkranz Richard R , Lubans David R , Peralta Louisa R , Bennie Andrew , Sanders Taren , Lonsdale Chris 
Published: October 2012

Abstract

Background
The physical activity (PA) levels of many children and adolescents in Australia are currently insufficient to promote health benefits. Physical education (PE) programs aim to promote PA and reach nearly all school-aged children, but PA levels within PE lessons are often low. PE teachers may influence children’s motivation to be physically active in PE lessons, but little is known about teacher strategies that effectively motivate children to participate in PA, and few intervention studies have examined motivational strategies in PE. The purpose of this study was to compare the effect of three motivational strategies, each based on Self-Determination Theory (SDT), on PA levels, and their hypothesized antecedents, during year 8 PE lessons.

Methods/design
This study employed a cluster-randomized controlled trial design. Following a familiarization session, PA levels and hypothesized PA antecedents were measured during a baseline lesson and a post-intervention or control lesson. Teachers (n = 16) and their classes from five secondary schools in Sydney, Australia were randomly assigned into four blocks and instructed to provide one of four 20-min lesson teaching strategy conditions: (1) explaining the relevance of activities; (2) providing choice from PA options selected by the teacher; (3) providing equipment and free choice of activities; or (4) usual practice. The primary outcomes were lesson time spent in moderate-to-vigorous PA, and motivation towards the lesson. Secondary outcomes were perceptions of teacher behavior, psychological needs satisfaction, and lesson time spent in sedentary behavior. PA and sedentary behavior were measured during baseline and post-intervention lessons with waist-mounted Actigraph GT3X accelerometers. Teacher behavior, psychological needs satisfaction, and motivation were assessed via questionnaires at the end of each lesson. Linear mixed-model analyses will be run on all outcomes, with students nested within teachers as a random effect.

Discussion
Study findings will inform the development of effective SDT-based teaching strategy interventions to enhance students’ psychological needs satisfaction, motivation, and PA levels. More effective teaching strategies may be identified that are capable of improving adolescents’ PA levels, and thereby provide beneficial population health outcomes.

Trial registration
This trial is registered with Current Controlled Trials and is traceable as ISRCTN07038258.
Homeopathy for mental fatigue: lessons from a randomized, triple blind, placebo-controlled cross-over clinical trial
Authors: Dean Michael Emmans , Karsandas Raj , Bland J Martin , Gooch Debbie , MacPherson Hugh 
Published: October 2012

Abstract

Background
Difficulty in controlling attention can lead to mental fatigue in the healthy population. We identified one trial reporting a benefit in patients’ attention using a homeopathic formula preparation. One component of the preparation was potassium phosphate, widely available off the shelf as Kali phos 6x for cognitive problems. The aim of this exploratory trial was to assess the effectiveness of Kali phos 6x for attention problems associated with mental fatigue.

Methods
We recruited student and staff volunteers (University of York) with self-reported mental fatigue, excluding any using homeopathy or prescribed stimulants, or with a diagnosis of chronic fatigue syndrome. In a triple blind, cross-over, placebo-controlled clinical trial, 86 volunteers were randomized to receive Kali phos 6x or identical placebo 10 minutes before taking a psychological test of attention (Stroop Colour-Word Test). One week later they were crossed over and took the other preparation before repeating the test.

Results
We found no evidence of a treatment effect in a comparison of Kali phos 6x with placebo (Kali phos minus placebo = −1.1 (95% CI −3.0 to 0.9, P = 0.3) Stroop score units, Cohen effect size = −0.17) even when allowing for a weak period effect with accuracy scores in the second period being higher than those in the first (P = 0.05). We observed a ceiling effect in the Stroop test which undermined our ability to interpret this result.

Conclusions

Kali phos 6x was not found to be effective in reducing mental fatigue. A ceiling effect in our primary outcome measure meant that we could not rule out a type II error. Thorough piloting of an adequate outcome measure could have led to an unequivocal result.

Current Controlled Trials
ISRCTN16521161
Frequency of subtype B and F1 dual infection in HIV-1 positive, Brazilian men who have sex with men
Authors: Soares de Oliveira Ana Carolina , Pessôa de Farias Rodrigo , da Costa Antonio Charlys , Sauer Mariana Melillo , Bassichetto Katia Cristina , Oliveira Solange Maria Santos , Costa Priscilla Ramos , Tomiyama Claudia , Tomiyama Helena Tomoko Iwashita , Sabino Ester Cerdeira , Kallas Esper Georges , Sanabani Sabri Saeed 
Published: October 2012

Abstract

Background
Because various HIV vaccination studies are in progress, it is important to understand how often inter- and intra-subtype co/superinfection occurs in different HIV-infected high-risk groups. This knowledge would aid in the development of future prevention programs. In this cross-sectional study, we report the frequency of subtype B and F1 co-infection in a clinical group of 41 recently HIV-1 infected men who have sex with men (MSM) in São Paulo, Brazil.

Methodology
Proviral HIV-1 DNA was isolated from subject's peripheral blood polymorphonuclear leukocytes that were obtained at the time of enrollment. Each subject was known to be infected with a subtype B virus as determined in a previous study. A small fragment of the integrase gene (nucleotide 4255–4478 of HXB2) was amplified by nested polymerase chain reaction (PCR) using subclade F1 specific primers. The PCR results were further confirmed by phylogenetic analysis. Viral load (VL) data were extrapolated from the medical records of each patient.

Results
For the 41 samples from MSM who were recently infected with subtype B virus, it was possible to detect subclade F1 proviral DNA in five patients, which represents a co-infection rate of 12.2%. In subjects with dual infection, the median VL was 5.3 × 104 copies/ML, whereas in MSM that were infected with only subtype B virus the median VL was 3.8 × 104 copies/ML (p > 0.8).

Conclusions
This study indicated that subtype B and F1 co-infection occurs frequently within the HIV-positive MSM population as suggested by large number of BF1 recombinant viruses reported in Brazil. This finding will help us track the epidemic and provide support for the development of immunization strategies against the HIV.
Identification of a conserved linear B-cell epitope in the M protein of porcine epidemic diarrhea virus
Authors: Zhang Zhibang , Chen Jianfei , Shi Hongyan , Chen Xiaojin , Shi Da , Feng Li , Yang Bin 
Published: October 2012

Abstract

Background
The major structural protein of coronaviruses, the membrane (M) protein, can elicit the formation of protective antibodies, but little information is available about the M protein of porcine epidemic diarrhea virus (PEDV). Identification of epitopes on the PEDV M protein will be helpful in the elucidation of the antigenic properties of this protein.

Results
One hybridoma cell line secreting anti-M protein monoclonal antibody (McAb) was generated and designated 4D4. To map the epitopes on the PEDV M protein, a total of 17 partially overlapping fragments covering the C-terminus of M protein were expressed as fusion proteins with a 6×His tag or a GST tag. A linear motif, 193TGWAFYVR200, was identified by enzyme-linked immunosorbent assay (ELISA) and western blot (WB) analysis using McAb 4D4. The motif 195WAFYVR200 was the minimal requirement for reactivity, as demonstrated by removing amino acids individually from both ends of the motif 193TGWAFYVR200. The result of WB analysis showed that the 4D4-defined epitope could be recognized by PEDV-positive serum, but not transmissible gastroenteritis virus (TGEV)-positive serum. Furthermore, this epitope was highly conserved among different PEDV strains, as shown by alignment and comparison of sequences.

Conclusion
A McAb, 4D4, directed against the M protein of PEDV, was obtained, and the 4D4-defined minimal epitope sequence was 195WAFYVR200. The McAb could serve as a candidate for development of a McAb-based antigen capture ELISA for detection of PEDV. The epitope identified provides a basis for the development of epitope-based differential diagnostic techniques and may be useful in the design of epitope-based vaccines.
Incidental intraoperative discovery of a pancreatic neuroendocrine tumor associated with chronic pancreatitis
Authors: Surlin Valeriu , Ramboiu Sandu , Ghilusi Mirela , Plesea Iancu Emil 
Published: October 2012

Abstract
Pancreatic neuroendocrine tumors are a rare entity with an incidence between 2 per million to 5 per 100 000. Association with pancreatitis (acute or chronic) is rare and is considered to be determined by the tumoral obstruction of pancreatic ducts, but sometimes occurs without any apparent relationship between them. Non-functional neuroendocrine pancreatic tumors are usually diagnosed when either very large or metastatic. Small ones are occasionally diagnosed when imagery is performed for other diagnostic reasons. Intraoperative discovery is even rarer and poses problems of differential diagnosis with other pancreatic tumors. Association with chronic pancreatitis is rare and usually due to pancreatic duct obstruction by the tumor. We describe the case of a patient with a small non-functioning neuroendocrine tumor in the pancreatic tail accidentally discovered during surgery for delayed traumatic splenic rupture associated with chronic alcoholic pancreatitis. The tumor of 1.5cm size was well differentiated and confined to the pancreas, and was resected by a distal splenopancreatectomy.

Conclusions

Surgeons should be well aware of the rare possibility of a non-functional neuroendocrine tumor in the pancreas, associated with chronic pancreatitis, surgical resection being the optimal treatment for cure. Histopathology is of utmost importance to establish the correct diagnosis, grade of differentiation, malignancy and prognosis.

Virtual slides

The virtual slide(s) for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/2114470176676003.
Reduction of peak plantar pressure in people with diabetes‐related peripheral neuropathy: an evaluation of the DH Pressure Relief Shoe™
Authors: Raspovic Anita , Landorf Karl B , Gazarek Jana , Stark Megan 
Published: October 2012

AbstractBackgroundOffloading plantar pressure is a key strategy for the prevention or healing of neuropathic plantar ulcers in diabetes. Non‐removable walking casts, such as total contact casts, are currently considered the gold‐standard for offloading this type of wound. However, alternative methods for offloading that are more cost effective and easier to use are continually being sought. The aim of this study was to evaluate the capacity of the DH Pressure Relief Shoe™ to offload high pressure areas under the neuropathic foot in diabetes.MethodsA within‐subjects, repeated measures design was used. Sixteen participants with diabetic peripheral neuropathy were recruited and three footwear conditions were evaluated in a randomised order: a canvas shoe (the control), the participants' own standard shoe, and the DH Pressure Relief Shoe™. The primary outcome was peak plantar pressure, measured using the pedar‐X® mobile in‐shoe system between the three conditions.ResultsData analysis was conducted on 14 out of the 16 participants because two participants could not complete data collection. The mean peak pressure values in kPa (±SD) for each condition were: control shoe 315.9 (±140.7), participants' standard shoe 273.0 (±127.1) and DH Pressure Relief Shoe™ 155.4 (±89.9). There was a statistically significant difference in peak plantar pressure between the DH Pressure Relief Shoe™ compared to both the control shoe (p = 0.002) and participants' standard shoe (p = 0.001). The DH Pressure Relief Shoe™ decreased plantar pressures by 51% compared to the control shoe and by 43% compared to participants' standard shoe. Importantly, for a couple of study participants, the DH Pressure Relief Shoe™ appeared unsuitable for day‐to‐day wearing.ConclusionsThe DH Pressure Relief Shoe™ reduced plantar pressures more than the other two shoe conditions. The DH Pressure Relief Shoe™ may be a useful alternative to current offloading modalities used in clinical management of diabetic foot ulceration. However, clinical trials are needed to test their effectiveness for ulcer healing and to ensure they are useable and safe for patients in everyday activities.
A positive serum basophil histamine release assay is a marker for ciclosporin‐responsiveness in patients with chronic spontaneous urticaria
Authors: Iqbal Kamran , Bhargava Kapil , Skov Per Stahl , Falkencrone Sidsel , Grattan Clive EH 
Published: October 2012

AbstractThe electronic records of 398 patients with chronic spontaneous urticaria (CSU) who had had a serum basophil histamine release assay (BHRA) performed as a marker of functional autoantibodies were audited. The BHRA was positive in 105 patients (26.4%). Fifty eight were treated with ciclosporin because they were H1 anti‐histamine unresponsive. CSU patients with a positive BHRA were more likely to respond clinically (P<0.001) and to have raised thyroid autoantibodies (P<0.02) than those with a negative BHRA. The BHRA offers a useful predictive biomarker for a good response of H1 antihistamine‐unresponsive CSU patients to ciclosporin.
Founder mutations and genotype-phenotype correlations in Meckel-Gruber syndrome and associated ciliopathies
Authors: Szymanska Katarzyna , Berry Ian , Logan Clare V , Cousins Simon RR , Lindsay Helen , Jafri Hussain , Raashid Yasmin , Malik-Sharif Saghira , Castle Bruce , Ahmed Mushtag , Bennett Chris , Carlton Ruth , Johnson Colin A 
Published: October 2012

Abstract

Background
Meckel-Gruber syndrome (MKS) is an autosomal recessive lethal condition that is a ciliopathy. MKS has marked phenotypic variability and genetic heterogeneity, with mutations in nine genes identified as causative to date.

Methods
Families diagnosed with Meckel-Gruber syndrome were recruited for research studies following informed consent. DNA samples were analyzed by microsatellite genotyping and direct Sanger sequencing.

Results
We now report the genetic analyses of 87 individuals from 49 consanguineous and 19 non-consanguineous families in an unselected cohort with reported MKS, or an associated severe ciliopathy in a kindred. Linkage and/or direct sequencing were prioritized for seven MKS genes (MKS1, TMEM216, TMEM67/MKS3, RPGRIP1L, CC2D2A, CEP290 and TMEM237) selected on the basis of reported frequency of mutations or ease of analysis. We have identified biallelic mutations in 39 individuals, of which 13 mutations are novel and previously unreported. We also confirm general genotype-phenotype correlations.

Conclusions

TMEM67 was the most frequently mutated gene in this cohort, and we confirm two founder splice-site mutations (c.1546 + 1 G > A and c.870-2A > G) in families of Pakistani ethnic origin. In these families, we have also identified two separate founder mutations for RPGRIP1L (c. 1945 C > T p.R649X) and CC2D2A (c. 3540delA p.R1180SfsX6). Two missense mutations in TMEM67 (c. 755 T > C p.M252T, and c. 1392 C > T p.R441C) are also probable founder mutations. These findings will contribute to improved genetic diagnosis and carrier testing for affected families, and imply the existence of further genetic heterogeneity in this syndrome.
How effective are slurry storage, cover or catch crops, woodland creation, controlled trafficking or break-up of compacted layers, and buffer strips as on-farm mitigation measures for delivering an improved water environment?
Authors: Randall Nicola P , Donnison Louise M , Lewis Paul J 
Published: October 2012

Abstract
Background
Agriculture has intensified over the last 50 years resulting in increased usage of fertilizers and agrochemicals, changes in cropping practices, land drainage and increased stocking rates. In Europe, this has resulted in declines in the quality of soils and waters due to increased run off and water pollution. Fifty percent of nitrates in European rivers are derived from agricultural sources in the UK this value is as high as 70%, where agriculture also contributes to approximately 28% of phosphates and 76% of sediments recorded in rivers. Catchments dominated by agricultural land use have increased levels of pesticides and bacterial pathogens.
European member states have a policy commitment to tackle water pollution through the Water Framework Directive. An analysis of the effectiveness of water pollution mitigation measures should enable decision makers and delivery agencies to better facilitate catchment planning.
The aim of this systematic review is to assess the effectiveness of slurry storage, cover/catch crops, woodland creation, controlled trafficking/break-up of compacted layers and buffer strips, as on farm mitigation measures, for delivering an improved water environment.

Methods
The systematic review will consist of a searchable systematic map database for all the named interventions. Where possible, quantitative analysis will be used to assess the effectiveness of interventions.
Electronic databases, the internet, and organisational websites will be searched, and stakeholders will be contacted for studies that investigate the impact of the on-farm mitigation measures on water quality. All studies found will be assessed for suitability for inclusion in the next stage. Inclusion criteria will be based on subject, intervention, comparator and outcome. The details of included studies will be incorporated into the systematic map database, and studies scored for effectiveness of intervention and study design. Where there is suitable data available, meta-analysis will be carried out to test the effectiveness of individual mitigation measures. A report will summarise the evidence, highlight any gaps in the available research, and provide recommendations for future research.
Decreased cervical epithelial sensitivity to nonoxynol-9 (N-9) after four daily applications in a murine model of topical vaginal microbicide safety
Authors: Lozenski Karissa , Ownbey Robert , Wigdahl Brian , Kish-Catalone Tina , Krebs Fred C 
Published: October 2012

Abstract

Background
The disappointing clinical failures of five topical vaginal microbicides have provided new insights into factors that impact microbicide safety and efficacy. Specifically, the greater risk for human immunodeficiency virus type 1 (HIV-1) acquisition associated with multiple uses of a nonoxynol-9 (N-9)-containing product has highlighted the importance of application frequency as a variable during pre-clinical microbicide development, particularly in animal model studies.

Methods
To evaluate an association between application frequency and N-9 toxicity, experiments were performed using a mouse model of cervicovaginal microbicide safety. In this model system, changes in cervical and vaginal epithelial integrity, cytokine release, and immune cell infiltration were assessed after single and multiple exposures to N-9.

Results
After the initial application of N-9 (aqueous, 1%), considerable damage to the cervical epithelium (but not the vaginal epithelium) was observed as early as 10 min post-exposure and up to 8 h post-exposure. Subsequent daily exposures (up to 4 days) were characterized by diminished cervical toxicity relative to single exposures of like duration. Levels of pro-inflammatory cytokines released into the cervicovaginal lumen and the degree of CD14-positive immune cell infiltration proximal to the cervical epithelium were also dependent on the number of N-9 exposures.

Conclusions
Rather than causing cumulative cervical epithelial damage, repeated applications of N-9 were characterized by decreased sensitivity to N-9-associated toxicity and lower levels of immune cell recruitment. These results provide new insights into the failure of N-9-based microbicides and illustrate the importance of considering multiple exposure protocols in pre-clinical microbicide development strategies.