bluuebunny/crossref_metadata_2025 huggingface dataset:
A new microspore embryogenesis system under low temperature which mimics zygotic embryogenesis initials, expresses auxin and efficiently regenerates doubled-haploid plants in Brassica napus
Authors: Prem Deepak , Solís María-Teresa , Bárány Ivett , Rodríguez-Sanz Héctor , Risueño María C , Testillano Pilar S
Published: August 2012

Background

Microspore embryogenesis represents a unique system of single cell reprogramming in plants wherein a highly specialized cell, the microspore, by specific stress treatment, switches its fate towards an embryogenesis pathway. InBrassica napus,a model species for this phenomenon, incubation of isolated microspores at 32°C is considered to be a pre-requisite for embryogenesis induction.

Results

We have developed a newin vitrosystem at lower temperature (18°C) to efficiently induce microspore embryogenesis throughout two different developmental pathways: one involving the formation of suspensor-like structures (52.4%) and another producing multicellular embryos without suspensor (13.1%); additionally, a small proportion of non-responsive microspores followed a gametophytic-like development (34.4%) leading to mature pollen. The suspensor-like pathway followed at 18°C involved the establishment of asymmetric identities from the first microspore division and an early polarity leading to different cell fates, suspensor and embryo development, which were formed by cells with different organizations and endogenous auxin distribution, similar to zygotic embryogenesis. In addition, a new strategy for germination of microspore derived embryos was developed for achieving more than 90% conversion of embryos to plantlets, with a predominance of spontaneous doubled haploids plants.

Conclusion

The present work reveals a novel mechanism for efficient microspore embryogenesis induction inB. napususing continuous low temperature treatment. Results indicated that low temperature applied for longer periods favours an embryogenesis pathway whose first division originates asymmetric cell identities, early polarity establishment and the formation of suspensor-like structures, mimicking zygotic embryogenesis. This newin vitrosystem provides a convenient tool to analyzein situthe mechanisms underlying different developmental pathways during the microspore reprogramming, breaking or not the cellular symmetry, the establishment of polarity and the developmental embryo patterning, which further produce mature embryos and plants.
Down-regulation of POLYGALACTURONASE1 alters firmness, tensile strength and water loss in apple (Malus x domestica) fruit
Authors: Atkinson Ross G , Sutherland Paul W , Johnston Sarah L , Gunaseelan Kularajathevan , Hallett Ian C , Mitra Deepali , Brummell David A , Schröder Roswitha , Johnston Jason W , Schaffer Robert J
Published: August 2012

Background

While there is now a significant body of research correlating apple (Malusxdomestica) fruit softening with the cell wall hydrolaseENDO-POLYGALACTURONASE1 (PG1), there is currently little knowledge of its physiological effectsin planta. This study examined the effect of down regulation of PG1 expression in ‘Royal Gala’ apples, a cultivar that typically has high levels of PG1, and softens during fruit ripening.

Results

PG1-suppressed ‘Royal Gala’ apples harvested from multiple seasons were firmer than controls after ripening, and intercellular adhesion was higher. Cell wall analyses indicated changes in yield and composition of pectin, and a higher molecular weight distribution of CDTA-soluble pectin. Structural analyses revealed more ruptured cells and free juice in pulled apart sections, suggesting improved integrity of intercellular connections and consequent cell rupture due to failure of the primary cell walls under stress.PG1-suppressed lines also had reduced expansion of cells in the hypodermis of ripe apples, resulting in more densely packed cells in this layer. This change in morphology appears to be linked with reduced transpirational water loss in the fruit.

Conclusions

These findings confirm PG1’s role in apple fruit softening and suggests that this is achieved in part by reducing cellular adhesion. This is consistent with previous studies carried out in strawberry but not with those performed in tomato. In apple PG1 also appears to influence other fruit texture characters such as juiciness and water loss.
Is the APLS formula used to calculate weight-for-age applicable to a Trinidadian population?
Authors: Ali Khalid , Sammy Ian , Nunes Paula
Published: August 2012

Background

In paediatric emergency medicine, estimation of weight in ill children can be performed in a variety of ways. Calculation using the ‘APLS’ formula (weight = [age + 4] × 2) is one very common method. Studies on its validity in developed countries suggest that it tends to under-estimate the weight of children, potentially leading to errors in drug and fluid administration. The formula is not validated in Trinidad and Tobago, where it is routinely used to calculate weight in paediatric resuscitation.

Methods

Over a six-week period in January 2009, all children one to five years old presenting to the Emergency Department were weighed. Their measured weights were compared to their estimated weights as calculated using the APLS formula, the Luscombe and Owens formula and a “best fit” formula derived (then simplified) from linear regression analysis of the measured weights.

Results

The APLS formula underestimated weight in all age groups with a mean difference of −1.4 kg (95% limits of agreement 5.0 to −7.8). The Luscombe and Owens formula was more accurate in predicting weight than the APLS formula, with a mean difference of −0.4 kg (95% limits of agreement 6.9 to −6.1%). Using linear regression analysis, and simplifying the derived equation, the best formula to describe weight and age was (weight = [2.5 x age] + 8). The percentage of children whose actual weight fell within 10% of the calculated weights using any of the three formulae was not significantly different.

Conclusions

The APLS formula slightly underestimates the weights of children in Trinidad, although this is less than in similar studies in developed countries. Both the Luscombe and Owens formula and the formula derived from the results of this study give a better estimate of the measured weight of children in Trinidad. However, the accuracy and precision of all three formulae were not significantly different from each other. It is recommended that the APLS formula should continue to be used to estimate the weight of children in resuscitation situations in Trinidad, as it is well known, easy to calculate and widely taught in this setting.
Immunophenotype classification and therapeutic outcomes of Chinese primary gastrointestinal diffuse large B-cell lymphoma
Authors: Zhang Zizhen , Shen Yanying , Shen Danping , Ni Xingzhi
Published: August 2012

Background

Recent studies showed that diffuse large B-cell lymphoma (DLBCL) could be classified into germinal centre B cell-like (GCB) and non-germinal centre B cell-like (non-GCB) phenotypes according to CD10,Bcl-6 and MUM1 expression. But primary gastrointestinal DLBCL has rarely been studied. This study was aimed to investigate the relationship between immunophenotypic classification, therapeutic outcomes and the prognosis of patients with primary gastrointestinal DLBCL.

Methods

Between 1998 and 2010, there were 151 patients studied at Shanghai Renji Hospital with a histopathological diagnosis of primary gastrointestinal DLBCL. Immunohistochemistry was performed using EnVision methods for CD10, BCL-6 and MUM1. The clinicopathologic features and follow-up data were analyzed by the Kaplan-Meier method, log-rank test and χ2 test.

Results

According to the expression of CD10, BCL-6 and MUM1, 31.8 % (48/151) of the cases belonged to the GCB subtype and 68.2 % (103/151) belonged to the non-GCB subtype. There was a significant difference of local lymph node metastasis between the GCB and non-GCB groups (P < 0.05). Patients in the GCB group had a better survival rate than those in the non-GCB group (5-year survival rate, 65.2 % vs 36.4 %, P < 0.05). In the GCB group, there was no significant difference in survival rates in patients receiving R-CHOP and CHOP therapy (P > 0.05). In the non-GCB group, the survival rate in patients treated with R-CHOP therapy was significantly longer than those treated with CHOP therapy (5-year survival rate, 62.8 % vs 30.8 %, P < 0.05).

Conclusions

The immunophenotype classification of gastrointestinal DLBCL, which is closely related to local lymph node metastasis, is found to have prognostic significance. Immunophenotype classification is also useful in selecting the chemotherapy protocol.
Evaluation of common genetic variants in 82 candidate genes as risk factors for neural tube defects
Authors: Pangilinan Faith , Molloy Anne M , Mills James L , Troendle James F , Parle-McDermott Anne , Signore Caroline , O’Leary Valerie B , Chines Peter , Seay Jessica M , Geiler-Samerotte Kerry , Mitchell Adam , VanderMeer Julia E , Krebs Kristine M , Sanchez Angelica , Cornman-Homonoff Joshua , Stone Nicole , Conley Mary , Kirke Peadar N , Shane Barry , Scott John M , Brody Lawrence C
Published: August 2012

Background

Neural tube defects (NTDs) are common birth defects (~1 in 1000 pregnancies in the US and Europe) that have complex origins, including environmental and genetic factors. A low level of maternal folate is one well-established risk factor, with maternal periconceptional folic acid supplementation reducing the occurrence of NTD pregnancies by 50-70%. Gene variants in the folate metabolic pathway (e.g., MTHFR rs1801133 (677 C > T) and MTHFD1 rs2236225 (R653Q)) have been found to increase NTD risk. We hypothesized that variants in additional folate/B12 pathway genes contribute to NTD risk.

Methods

A tagSNP approach was used to screen common variation in 82 candidate genes selected from the folate/B12 pathway and NTD mouse models. We initially genotyped polymorphisms in 320 Irish triads (NTD cases and their parents), including 301 cases and 341 Irish controls to perform case–control and family based association tests. Significantly associated polymorphisms were genotyped in a secondary set of 250 families that included 229 cases and 658 controls. The combined results for 1441 SNPs were used in a joint analysis to test for case and maternal effects.

Results

Nearly 70 SNPs in 30 genes were found to be associated with NTDs at the p < 0.01 level. The ten strongest association signals (p-value range: 0.0003–0.0023) were found in nine genes (MFTC, CDKN2A, ADA, PEMT, CUBN, GART, DNMT3A, MTHFD1 and T (Brachyury)) and included the known NTD risk factor MTHFD1 R653Q (rs2236225). The single strongest signal was observed in a new candidate, MFTC rs17803441 (OR = 1.61 [1.23-2.08], p = 0.0003 for the minor allele). Though nominally significant, these associations did not remain significant after correction for multiple hypothesis testing.

Conclusions

To our knowledge, with respect to sample size and scope of evaluation of candidate polymorphisms, this is the largest NTD genetic association study reported to date. The scale of the study and the stringency of correction are likely to have contributed to real associations failing to survive correction. We have produced a ranked list of variants with the strongest association signals. Variants in the highest rank of associations are likely to include true associations and should be high priority candidates for further study of NTD risk.
Deletion of a single-copy DAAM1 gene in congenital heart defect: a case report
Authors: Bao Bihui , Zhang Liang , Hu Hua , Yin Shuxin , Liang Zhiqing
Published: August 2012

Background

With an increasing incidence of congenital heart defects (CHDs) in recent years, genotype-phenotype correlation and array-based methods have contributed to the genome-wide analysis and understanding of genetic variations in the CHD population. Here, we report a copy number deletion of chromosomal 14q23.1 in a female fetus with complex congenital heart defects. This is the first description of DAAM1 gene deletion associated with congenital heart anomalies.

Case Presentation

Compared with the control population, one CHD fetus showed a unique copy number deletion of 14q23.1, a region that harbored DAAM1 and KIAA0666 genes.

Conclusions

Results suggest that the copy number deletion on chromosome 14q23.1 may be critical for cardiogenesis. However, the exact relationship and mechanism of how DAAM1 and KIAA0666 deletion contributes to the onset of CHD is yet to be determined.
Investigating the relationship between UMODL1 gene polymorphisms and high myopia: a case–control study in Chinese
Authors: Zhu Miao-miao , Yap Maurice KH , Ho Daniel WH , Fung Wai Yan , Ng Po Wah , Gu Yang-shun , Yip Shea Ping
Published: August 2012

Background

The UMODL1 gene was found to be associated with high myopia in Japanese. This study aimed to investigate this gene for association with high myopia in Chinese.

Methods

Two groups of unrelated Han Chinese from Hong Kong were recruited using the same criteria: Sample Set 1 comprising 356 controls (spherical equivalent, SE, within ±1 diopter or D) and 356 cases (SE ≤ −8D), and Sample Set 2 comprising 394 controls and 526 cases. Fifty-nine tag single nucleotide polymorphisms (SNPs) were selected and genotyped for Sample Set 1. Four SNPs were followed up with Sample Set 2. Both single-marker and haplotype analyses were performed with cases defined by different SE thresholds. Secondary phenotypes were also analyzed for association with genotypes.

Results

Data filtering left 57 SNPs for analysis. Single-marker analysis did not reveal any significant differences between cases and controls in the initial study. However, haplotype GCT for markers rs220168-rs220170-rs11911271 showed marginal significance (empirical P = 0.076; SE ≤ −12D for cases), but could not be replicated in the follow-up study. In contrast, non-synonymous SNP rs3819142 was associated with high myopia (SE ≤ −10D) in the follow-up study, but could not be confirmed using Sample Set 1. The SNP rs2839471, positive in the original Japanese study, gave negative results in all our analyses. Exploratory analysis of secondary phenotypes indicated that allele C of rs220120 was associated with anterior chamber depth (adjusted P = 0.0460).

Conclusions

Common UMODL1 polymorphisms were unlikely to be important in the genetic susceptibility to high myopia in Han Chinese.
Expression of S100A4, ephrin-A1 and osteopontin in non-small cell lung cancer
Authors: Kongsgaard Ane , Lund-Iversen Marius , Berge Gisle , Brustugun Odd Terje , Solberg Steinar K , Mælandsmo Gunhild M , Boye Kjetil
Published: August 2012

Background

The metastasis-promoting protein S100A4 induces expression of ephrin-A1 and osteopontin in osteosarcoma cell lines. The aim of this study was to investigate S100A4-mediated stimulation of ephrin-A1 and osteopontin in non-small cell lung cancer (NSCLC) cell lines, and to characterize the expression of these biomarkers in primary tumor tissue from NSCLC patients.

Methods

Four NSCLC cell lines were treated with extracellular S100A4, and ephrin-A1 and osteopontin expression was analyzed by real time RT-PCR and Western blotting. Immunohistochemical staining for S100A4, ephrin-A1 and osteopontin was performed on tissue microarrays containing primary tumor samples from a cohort of 217 prospectively recruited NSCLC patients, and associations with clinicopathological parameters were investigated.

Results

S100A4 induced ephrin-A1 mRNA and protein expression in adenocarcinoma, but not in squamous carcinoma cell lines, whereas the level of osteopontin was unaffected by S100A4 treatment. In primary tumors, moderate or strong immunoreactivity was observed in 57% of cases for cytoplasmic S100A4, 46% for nuclear S100A4, 86% for ephrin-A1 and 77% for osteopontin. Interestingly, S100A4 expression was associated with ephrin-A1 also in vivo, but there was no association between S100A4 and osteopontin. Expression levels of S100A4 and ephrin-A1 were significantly higher in adenocarcinomas compared to other histological subtypes, and S100A4-positive tumors were smaller and more differentiated than tumors without expression.

Conclusions

Our findings suggest that S100A4, ephrin-A1 and osteopontin are involved in the biology of NSCLC, and further investigation of their potential use as biomarkers in NSCLC is warranted.
An association between hOGG1Ser326Cys polymorphism and the risk of bladder cancer in non-smokers: a meta-analysis
Authors: Ji Changwei , Liu Zhao , Chen Huimei , Guo Hongqian , Liu Changjian
Published: August 2012

Background

Bladder cancer results from complex interactions between many genetic and environment factors. The polymorphism Ser326Cys in hOGG1 gene has been reported to be associated with bladder cancer in some studies, though the results remain inconclusive. To explore this relationship of hOGG1 polymorphism and the susceptibility for bladder cancer and the impact of smoking exposures, a cumulative meta-analysis was performed in this study.

Methods

We extracted the data from the Pubmed database up to January 9, 2012 using the search phrases “hOGG1, Ser326Cys polymorphism and bladder cancer”. Seven case–control studies were identified, including 2474 patients and 2408 controls. Four of them provided the analysis of smoking effects, with 1372 smokers and 947 non-smokers. The odds ratios (ORs) and associated 95 % confidence intervals (CIs) were calculated using fixed- or random- effects models.

Results

Regarding the overall association between the hOGG1 326Cys allele and bladder cancer risk, the meta-analysis did not reveal a significant effect in the additive model (OR: 1.06, 95 % CI: 0.96-1.26; p = 0.49), the recessive genetic model (OR: 1.05, 95 % CI: 0.65-1.70; p = 0.85) or the dominant genetic model (OR: 1.07, 95 % CI: 0.87-1.32; p = 0.53). Similarly, no significant relationship was observed in the stratified analysis by ethnicity, study design and Hardy-Weinberg equilibrium (all p > 0.05). In the non-smokers, however, hOGG1 326Cys allele significantly increased the risk for bladder cancer and the ORs in the additive model, homozygote contrast and recessive genetic model were 1.59 (p = 0.02), 2.53(p = 0.003) and 2.41(p = 0.0005), respectively. Nevertheless, in the smoker subgroup, similar findings could not be found in all genetic models (all p > 0.05).

Conclusions

The association between the hOGG1 326Cys allele and bladder cancer was significant in non-smoker population, while was non-detectable in common or smoker populations. This meta-analysis suggests that the hOGG1 Ser326Cys polymorphism may be a risk factor for bladder cancer without exposure to smoking. Further functional studies are needed to elucidate the gene polymorphism-bladder cancer relationship and gene-environment interactions.
Snail transcription factor negatively regulates maspin tumor suppressor in human prostate cancer cells
Authors: Neal Corey L , Henderson Veronica , Smith Bethany N , McKeithen Danielle , Graham Tisheeka , Vo Baohan T , Odero-Marah Valerie A
Published: August 2012

Background

Maspin, a putative tumor suppressor that is down-regulated in breast and prostate cancer, has been associated with decreased cell motility. Snail transcription factor is a zinc finger protein that is increased in breast cancer and is associated with increased tumor motility and invasion by induction of epithelial-mesenchymal transition (EMT). We investigated the molecular mechanisms by which Snail increases tumor motility and invasion utilizing prostate cancer cells.

Methods

Expression levels were analyzed by RT-PCR and western blot analyses. Cell motility and invasion assays were performed, while Snail regulation and binding to maspin promoter was analyzed by luciferase reporter and chromatin immunoprecipitation (ChIP) assays.

Results

Snail protein expression was higher in different prostate cancer cells lines as compared to normal prostate epithelial cells, which correlated inversely with maspin expression. Snail overexpression in 22Rv1 prostate cancer cells inhibited maspin expression and led to increased migration and invasion. Knockdown of Snail in DU145 and C4-2 cancer cells resulted in up-regulation of maspin expression, concomitant with decreased migration. Transfection of Snail into 22Rv1 or LNCaP cells inhibited maspin promoter activity, while stable knockdown of Snail in C4-2 cells increased promoter activity. ChIP analysis showed that Snail is recruited to the maspin promoter in 22Rv1 cells.

Conclusions

Overall, this is the first report showing that Snail can negatively regulate maspin expression by directly repressing maspin promoter activity, leading to increased cell migration and invasion. Therefore, therapeutic targeting of Snail may be useful to re-induce expression of maspin tumor suppressor and prevent prostate cancer tumor progression.
The development of the Be Active & Relax “Vitality in Practice” (VIP) project and design of an RCT to reduce the need for recovery in office employees
Authors: Coffeng Jennifer K , Hendriksen Ingrid JM , Duijts Saskia F , Proper Karin I , van Mechelen Willem , Boot Cécile R L
Published: August 2012

Background

There is strong evidence to suggest that multiple work-related health problems are preceded by a higher need for recovery. Physical activity and relaxation are helpful in decreasing the need for recovery. This article aims to describe (1) the development and (2) the design of the evaluation of a daily physical activity and relaxation intervention to reduce the need for recovery in office employees.

Methods/Design

The study population will consist of employees of a Dutch financial service provider. The intervention was systematically developed, based on parts of the Intervention Mapping (IM) protocol. Assessment of employees needs was done by combining results of face-to-face interviews, a questionnaire and focus group interviews. A set of theoretical methods and practical strategies were selected which resulted in an intervention program consisting of Group Motivational Interviewing (GMI) supported by a social media platform, and environmental modifications. The Be Active & Relax program will be evaluated in a modified 2 X 2 factorial design. The environmental modifications will be pre-stratified and GMI will be randomised on department level. The program will be evaluated, using 4 arms: (1) GMI and environmental modifications; (2) environmental modifications; (3) GMI; (4) no intervention (control group). Questionnaire data on the primary outcome (need for recovery) and secondary outcomes (daily physical activity, sedentary behaviour, relaxation/detachment, work- and health-related factors) will be gathered at baseline (T0), at 6 months (T1), and at 12 months (T2) follow-up. In addition, an economic and a process evaluation will be performed.

Discussion

Reducing the need for recovery is hypothesized to be beneficial for employees, employers and society. It is assumed that there will be a reduction in need for recovery after 6 months and 12 months in the intervention group, compared to the control group. Results are expected in 2013.

Trial registration

Netherlands Trial Register (NTR): NTR2553
Sexually transmitted infections associated syndromes assisted in the primary health care in Northeast, Brazil
Authors: Cavalcante Elani Graça Ferreira , Araújo Maria Alix Leite , Galvão Marli Teresinha Gimeniz , de Moura Heber José , Gondim Ana Paula Soares , da Silva Raimunda Magalhães
Published: August 2012

Background

The lack of information on the care for sexually transmitted infections (STI) associated syndromes may contribute for its non-inclusion as prevention and control strategy for STI in Brazil. This study aims to analyze the cases of STI – Associated Syndromes assisted in primary health care center in a city in Northeast Brazil associating them with socio-demographic and behavioral variables.

Methods

This is a retrospective study that analyzed 5148 consultation forms and medical records of patients assisted in a primary health care center who presented at least one genital syndrome from 1999 to 2008. Was considered as dependent variables the genital syndromes and serologies for syphilis and HIV and as independent variables the socio-demographic and behavioral aspects. It was used Pearson’s chi-square test to analyze the differences between the categorical variables, with a significance level of 5%. It was performed a multivariate analysis through the multivariate logistic regression model with the variables with p <0.05. We used odds ratio with a confidence interval of 95%.

Results

The most frequent syndromes were vaginal discharge and/or cervicitis (44%) and genital wart (42.2%). Most people were between 20 and 39 years old (70%) and women (74.2%). Genital ulcer was most prevalent among men (OR = 2.67; CI 95% 1.99-3.58) and people who studied more than eight years (OR = 1.33; CI 95% 1.00-1.75) and wart prevailed among men (OR = 3.92; IC 95% 3.36-4.57), people under 29 years old (OR = 1.81; CI 95% 1.59-2.07) and who studied more than eight years (OR = 1.75; CI 95% 1.54-1.99). The Venereal Disease Research Laboratory (VDRL) was positive in 7.3% of men and in 7.1% of women and the Anti-HIV in 3.1% of men and 0.7% of women.

Conclusion

Vaginal discharge was the most frequent syndrome assisted in primary health care, followed by genital wart. The high prevalence of genital wart justifies the greater effort for the proper follow-up of these cases. Men presented more genital wart and ulcer and reported having more sexual partners, showing their need for a greater access and inclusion in health activities developed in primary health care in Brazil.
The gender gap in mobility: A global cross-sectional study
Authors: Mechakra-Tahiri Samia Djemâa , Freeman Ellen E , Haddad Slim , Samson Elodie , Zunzunegui Maria Victoria
Published: August 2012

Background

Several studies have demonstrated that women have greater mobility disability than men. The goals of this research were: 1) to assess the gender gap in mobility difficulty in 70 countries; 2) to determine whether the gender gap is explained by sociodemographic and health factors; 3) to determine whether the gender gap differs across 6 regions of the world with different degrees of gender equality according to United Nations data.

Methods

Population-based data were used from the World Health Survey (WHS) conducted in 70 countries throughout the world. 276,647 adults aged 18 years and over were recruited from 6 world regions. Mobility was measured by asking the level of difficulty people had moving around in the last 30 days and then creating a dichotomous measure (no difficulty, difficulty). The human development index and the gender-related development index for each country were obtained from the United Nations Development Program website. Poisson regression with Taylor series linearized variance estimation was used.

Results

Women were more likely than men to report mobility difficulty (38% versus 27%, P < 0.0001). The age-adjusted prevalence rate ratio for female gender was 1.35 (95% CI 1.31–1.38). The addition of education, marital status, and urban versus rural setting reduced the prevalence rate ratio to 1.30 (95% CI 1.26–1.33). The addition of the presence of back pain, arthritis, angina, depressive symptoms, and cognitive difficulties further reduced the prevalence rate ratio to 1.12 (95% CI 1.09–1.15). There was statistical interaction on the multiplicative scale between female gender and region (P < 0.01). The Eastern Mediterranean region, which had the greatest loss of human development due to gender inequality, showed the largest gender gap in mobility difficulty, while the Western Pacific region, with the smallest loss of human development due to gender inequality, had the smallest gender gap in mobility difficulty.

Conclusions

These are the first world-wide data to examine the gender gap in mobility. Differences in chronic diseases are the main reasons for this gender gap. The gender gap seems to be greater in regions with the largest loss of human development due to gender inequality.
Development of a transient expression assay for detecting environmental oestrogens in zebrafish and medaka embryos
Authors: Lee Okhyun , Tyler Charles R , Kudoh Tetsuhiro
Published: August 2012

Background

Oestrogenic contaminants are widespread in the aquatic environment and have been shown to induce adverse effects in both wildlife (most notably in fish) and humans, raising international concern. Available detecting and testing systems are limited in their capacity to elucidate oestrogen signalling pathways and physiological impacts. Here we developed a transient expression assay to investigate the effects of oestrogenic chemicals in fish early life stages and to identify target organs for oestrogenic effects. To enhance the response sensitivity to oestrogen, we adopted the use of multiple tandem oestrogen responsive elements (EREc38) in a Tol2 transposon mediated Gal4ff-UAS system. The plasmid constructed (pTol2_ERE-TATA-Gal4ff), contains three copies of oestrogen response elements (3ERE) that on exposure to oestrogen induces expression of Gal4ff which this in turn binds Gal4-responsive Upstream Activated Sequence (UAS) elements, driving the expression of a second reporter gene, EGFP (Enhanced Green Fluorescent Protein).

Results

The response of our construct to oestrogen exposure in zebrafish embryos was examined using a transient expression assay. The two plasmids were injected into 1–2 cell staged zebrafish embryos, and the embryos were exposed to various oestrogens including the natural steroid oestrogen 17ß-oestradiol (E2), the synthetic oestrogen 17α- ethinyloestradiol (EE2), and the relatively weak environmental oestrogen nonylphenol (NP), and GFP expression was examined in the subsequent embryos using fluorescent microscopy. There was no GFP expression detected in unexposed embryos, but specific and mosaic expression of GFP was detected in the liver, heart, somite muscle and some other tissue cells for exposures to steroid oestrogen treatments (EE2; 10 ng/L, E2; 100 ng/L, after 72 h exposures). For the NP exposures, GFP expression was observed at 10 μg NP/L after 72 h (100 μg NP/L was toxic to the fish). We also demonstrate that our construct works in medaka, another model fish test species, suggesting the transient assay is applicable for testing oestrogenic chemicals in fish generally.

Conclusion

Our results indicate that the transient expression assay system can be used as a rapid integrated testing system for environmental oestrogens and to detect the oestrogenic target sites in developing fish embryos.
Profiling the clinical presentation of diagnostic characteristics of a sample of symptomatic TMD patients
Authors: e Silva Machado Luciana Pimenta , de Macedo Nery Marianita Batista , de Góis Nery Cláudio , Leles Cláudio Rodrigues
Published: August 2012

Background

Temporomandibular disorder (TMD) patients might present a number of concurrent clinical diagnoses that may be clustered according to their similarity. Profiling patients’ clinical presentations can be useful for better understanding the behavior of TMD and for providing appropriate treatment planning. The aim of this study was to simultaneously classify symptomatic patients diagnosed with a variety of subtypes of TMD into homogenous groups based on their clinical presentation and occurrence of comorbidities.

Methods

Clinical records of 357 consecutive TMD patients seeking treatment in a private specialized clinic were included in the study sample. Patients presenting multiple subtypes of TMD diagnosed simultaneously were categorized according to the AAOP criteria. Descriptive statistics and two-step cluster analysis were used to characterize the clinical presentation of these patients based on the primary and secondary clinical diagnoses.

Results

The most common diagnoses were localized masticatory muscle pain (n = 125) and disc displacement without reduction (n = 104). Comorbidity was identified in 288 patients. The automatic selection of an optimal number of clusters included 100% of cases, generating an initial 6-cluster solution and a final 4-cluster solution. The interpretation of within-group ranking of the importance of variables in the clustering solutions resulted in the following characterization of clusters: chronic facial pain (n = 36), acute muscle pain (n = 125), acute articular pain (n = 75) and chronic articular impairment (n = 121).

Conclusion

Subgroups of acute and chronic TMD patients seeking treatment can be identified using clustering methods to provide a better understanding of the clinical presentation of TMD when multiple diagnosis are present. Classifying patients into identifiable symptomatic profiles would help clinicians to estimate how common a disorder is within a population of TMD patients and understand the probability of certain pattern of clinical complaints.
Reduced Newcastle disease virus-induced oncolysis in a subpopulation of cisplatin-resistant MCF7 cells is associated with survivin stabilization
Authors: Jamal Mohd-Hafifi , Ch’ng Wei-Choong , Yusoff Khatijah , Shafee Norazizah
Published: August 2012

Background

Cisplatin resistance is a serious problem in cancer treatment. To overcome it, alternative approaches including virotherapy are being pursued. One of the candidates for anticancer virotherapy is the Newcastle disease virus (NDV). Even though NDV's oncolytic properties in various cancer cells have been widely reported, information regarding its effects on cisplatin resistant cancer cells is still limited. Therefore, we tested the oncolytic efficacy of a strain of NDV, designated as AF2240, in a cisplatin-resistant breast cancer cell line.

Methods

Cisplatin-resistant cell line (MCF7-CR) was developed from the MCF7 human breast adenocarcinoma cell line by performing a seven-cyclic exposure to cisplatin. Following NDV infection, fluorescence-activated cell sorting (FACS) analysis and immunoblotting were used to measure cell viability and viral protein expression, respectively. Production of virus progeny was then assessed by using the plaque assay technique.

Results

Infection of a mass population of the MCF7-CR with NDV resulted in 50% killing in the first 12 hours post-infection (hpi), comparable to the parental MCF7. From 12 hpi onwards, the remaining MCF7-CR became less susceptible to NDV killing. This reduced susceptibility led to increased viral protein synthesis and virus progeny production. The reduction was also associated with a prolonged cell survival via stabilization of the survivin protein.

Conclusions

Our findings showed for the first time, the involvement of survivin in the reduction of NDV-induced oncolysis in a subpopulation of cisplatin-resistant cells. This information will be important towards improving the efficacy of NDV as an anticancer agent in drug resistant cancers.
Clinical tolerability of artesunate-amodiaquine versus comparator treatments for uncomplicated falciparum malaria: an individual-patient analysis of eight randomized controlled trials in sub-Saharan Africa
Authors: Zwang Julien , Dorsey Grant , Djimdé Abdoulaye , Karema Corine , Mårtensson Andreas , Ndiaye Jean-Louis , Sirima Sodiomon B , Olliaro Piero
Published: August 2012

Background

The widespread use of artesunate-amodiaquine (ASAQ) for treating uncomplicated malaria makes it important to gather and analyse information on its tolerability.

Methods

An individual-patient tolerability analysis was conducted using data from eight randomized controlled clinical trials conducted at 17 sites in nine sub-Saharan countries comparing ASAQ to other anti-malarial treatments. All patients who received at least one dose of the study drug were included in the analysis. Differences in adverse event (AE) and treatment emergent adverse event (TEAE) were analysed by Day 28.

Results

Of the 6,179 patients enrolled (74% <5 years of age), 50% (n = 3,113) received ASAQ, 20% (n = 1,217) another ACT, and 30% (n = 1,849) a non-ACT (combination or single-agent) treatment. Overall, 8,542 AEs were recorded. The proportion of patients experiencing at least one gastro-intestinal AE on ASAQ was 43% (and higher than that with artemether-lumefantrine and dihydroartemisinin-piperaquine at two sites), and was 23% for any other AEs (not different from other treatments). Specifically, the risk of diarrhoea, vomiting, cough and weakness was lower with artemether-lumefantrine; artemether-lumefantrine and dihydroartemisinin-piperaquine carried a higher risk of pruritus, chloroquine-SP had a higher risk of nausea. Parasitological recurrence increased the risk of occurrence of any AE. No other difference was detected. Comparing AE to TEAE in patients who had pre-treatment occurrence and grades of intensity recorded, AEs were significantly more related to the pre-treatment prevalence of the symptom (p = 0.001, Fisher test); AEs overestimated TEAEs by a factor ranging from none to five-fold. The overall incidence of serious AEs (SAEs) with ASAQ was nine per 1,000 (29/3,113) and mortality was one per 1,000 (three deaths, none drug-related); both were similar to other treatments.

Conclusion

ASAQ was comparatively well-tolerated. Safety information is important, and must be collected and analysed in a standardized way. TEAEs are a more objective measure of treatment-induced toxicity.
ATP driven clathrin dependent entry of carbon nanospheres prefer cells with glucose receptors
Authors: Selvi Ruthrotha B , Chatterjee Snehajyoti , Jagadeesan Dinesh , Chaturbedy Piyush , Suma Bangalore Srinivas , Eswaramoorthy Muthusamy , Kundu Tapas K
Published: August 2012

Background

Intrinsically fluorescent glucose derived carbon nanospheres (CSP) efficiently enter mammalian cells and also cross the blood brain barrier (BBB). However, the mechanistic details of CSP entry inside mammalian cells and its specificity are not known.

Results

In this report, the biochemical and cellular mechanism of CSP entry into the living cell have been investigated. By employing confocal imaging we show that CSP entry into the mammalian cells is an ATP-dependent clathrin mediated endocytosis process. Zeta potential studies suggest that it has a strong preference for cells which possess high levels of glucose transporters such as the glial cells, thereby enabling it to target individual organs/tissues such as the brain with increased specificity.

Conclusion

The endocytosis of Glucose derived CSP into mammalian cells is an ATP dependent process mediated by clathrin coated pits. CSPs utilize the surface functional groups to target cells containing glucose transporters on its membrane thereby implicating a potential application for specific targeting of the brain or cancer cells.
A scoping review about conference objectives and evaluative practices: how do we get more out of them?
Authors: Neves Justin , Lavis John N , Ranson M Kent
Published: August 2012

Large multi-day conferences have often been criticized as ineffective ways to improve social outcomes and to influence policy or practice. Unfortunately, many conference evaluations have also been inadequate in determining the impact of a conference on its associated social sector, with little evidence gathered or analyzed to substantiate or refute these criticisms. The aim of this scoping review is to investigate and report stakeholders’ objectives for planning or participating in large multi-day conferences and how these objectives are being evaluated. We conducted a scoping review supplemented by a small number of key informant interviews. Eight bibliographic databases were systematically searched to identify papers describing conference objectives and/or evaluations. We developed a conference evaluation framework based on theoretical models and empirical findings, which structured the descriptive synthesis of the data. We identified 3,073 potential papers for review, of which 44 were included in this study. Our evaluation framework connects five key elements in planning a conference and its evaluation (number in brackets refers to number of themes identified): conference objectives (8), purpose of evaluation (7), evaluation methods (5), indicators of success (9) and theories/models (8). Further analysis of indicators of success identified three categories of indicators with differing scopes (i.e. immediate, prospective or follow-up) as well as empirical links between the purpose of evaluations and these indicators. Conference objectives and evaluations were largely correlated with the type of conference (i.e. academic, political/governmental or business) but diverse overall. While much can be done to improve the quality and usefulness of conference evaluations, there are innovative assessments that are currently being utilized by some conferences and warrant further investigation. This review provides conference evaluators and organizers a simple resource to improve their own assessments by highlighting and categorizing potential objectives and evaluation strategies.
Serum microRNAs as biomarkers for recurrence in melanoma
Authors: Friedman Erica B , Shang Shulian , de Miera Eleazar Vega-Saenz , Fog Jacob Ulrik , Teilum Maria Wrang , Ma Michelle W , Berman Russell S , Shapiro Richard L , Pavlick Anna C , Hernando Eva , Baker Adam , Shao Yongzhao , Osman Iman
Published: August 2012

Background

Identification of melanoma patients at high risk for recurrence and monitoring for recurrence are critical for informed management decisions. We hypothesized that serum microRNAs (miRNAs) could provide prognostic information at the time of diagnosis unaccounted for by the current staging system and could be useful in detecting recurrence after resection.

Methods

We screened 355 miRNAs in sera from 80 melanoma patients at primary diagnosis (discovery cohort) using a unique quantitative reverse transcription-PCR (qRT-PCR) panel. Cox proportional hazard models and Kaplan-Meier recurrence-free survival (RFS) curves were used to identify a miRNA signature with prognostic potential adjusting for stage. We then tested the miRNA signature in an independent cohort of 50 primary melanoma patients (validation cohort). Logistic regression analysis was performed to determine if the miRNA signature can determine risk of recurrence in both cohorts. Selected miRNAs were measured longitudinally in subsets of patients pre-/post-operatively and pre-/post-recurrence.

Results

A signature of 5 miRNAs successfully classified melanoma patients into high and low recurrence risk groups with significant separation of RFS in both discovery and validation cohorts (p = 0.0036, p = 0.0093, respectively). Significant separation of RFS was maintained when a logistic model containing the same signature set was used to predict recurrence risk in both discovery and validation cohorts (p < 0.0001, p = 0.033, respectively). Longitudinal expression of 4 miRNAs in a subset of patients was dynamic, suggesting miRNAs can be associated with tumor burden.

Conclusion

Our data demonstrate that serum miRNAs can improve accuracy in identifying primary melanoma patients with high recurrence risk and in monitoring melanoma tumor burden over time.
Spectroscopic study on the interaction of pristine C60 and serum albumins in solution
Authors: Liu Shufang , Sui Yu , Guo Kai , Yin Zhijuan , Gao Xibao
Published: August 2012

The interaction of nanomaterials with biological macromolecules is an important foundation of the design and the biological safety assessments of nanomaterials. This work aims to investigate the interaction between pristine C60 and serum albumins (human serum albumin and bovine serum albumin) in solution. Stable aqueous dispersion of C60 was prepared by simple direct ultrasonic method and characterized by UV–vis spectrophotometry, transmission electronic microscopy and dynamic light scattering techniques, and spectroscopic methods (fluorescence spectroscopy, synchronous fluorescence spectroscopy and circular dichroism spectroscopy) were utilized for the investigation. It was found that the fluorescence of serum albumins could be quenched by C60 nanoparticles in a substantially similar way. Slight changes of the surrounding microenvironment of amino residues were observed, while little effects on the protein secondary structure occurred. The different effects of dispersion methods on the interaction of C60 nanoparticles with serum protein were also compared and discussed.
Municipal solid waste landfill leachate treatment by fenton, photo-fenton and fenton-like processes: Effect of some variables
Authors: Zazouli Mohammad Ali , Yousefi Zabihollah , Eslami Akbar , Ardebilian Maryam Bagheri
Published: August 2012

Advanced oxidation processes like Fenton and photo-Fenton have been effectively applied to oxidize the persistent organic compounds in solid waste leachate and convert them to unharmful materials and products. However, there are limited data about application of Fenton-like process in leachate treatment. Therefore, this study was designed with the objective of treating municipal landfill leachate by Fenton, Fenton-like and photo–Fenton processes to determine the effect of different variables, by setting up a pilot system. The used leachate was collected from a municipal unsanitary landfill in Qaem-Shahr in the north of Iran. Fenton and Fenton-like processes were conducted by Jar-test method. Photo-Fenton process was performed in a glass photo-reactor. In all processes, H2O2was used as the oxidant. FeSO4.7H2O and FeCl3.6H2O were used as reagents. All parameters were measured based on standard methods. The results showed that the optimum concentration of H2O2was equal to 5 g/L for the Fenton-like process and 3 g/L for the Fenton and photo-Fenton processes. The optimum ratio of H2O2: Fe+2/Fe+3were equal to 8:1 in all processes. At optimum conditions, the amount of COD removal was 69.6%, 65.9% and 83.2% in Fenton, Fenton-like and photo–Fenton processes, respectively. In addition, optimum pH were 3, 5 and 3 and the optimum contact time were 150, 90 and 120 minutes, for Fenton, Fenton-like and photo–Fenton processes, respectively. After all processes, the biodegradability (BOD5/COD ratio) of the treated leachate was increased compared to that of the raw leachate and the highest increase in BOD5/COD ratio was observed in the photo-Fenton process. The efficiency of the Fenton-like process was overally less than Fenton and photo-Fenton processes, meanwhile the Fenton-like process was at higher pH and did not show problems.
Sludge reduction by lumbriculus variegatusin Ahvas wastewater treatment plant
Authors: Basim Yalda , Farzadkia Mahdi , Jaafarzadeh Nematollah , Hendrickx Tim
Published: August 2012

Sludge production is an avoidable problem arising from the treatment of wastewater. The sludge remained after municipal wastewater treatment contains considerable amounts of various contaminants and if is not properly handled and disposed, it may produce extensive health hazards. Application of aquatic worm is an approach to decrease the amount of biological waste sludge produced in wastewater treatment plants. In the present research reduction of the amount of waste sludge from Ahvaz wastewater treatment plant was studied with the aquatic worm Lumbriculus variegatus in a reactor concept. The sludge reduction in the reactor with worm was compared to sludge reduction in a blank reactor (without worm). The effects of changes in dissolved oxygen (DO) concentration up to 3 mg/L (run 1) and up to 6 mg/L (run 2) were studied in the worm and blank reactors. No meaningful relationship was found between DO concentration and the rate of total suspended solids reduction. The average sludge reductions were obtained as 32% (run 1) and 33% (run 2) in worm reactor and 16% (run 1) and 12% (run 2) in the blank reactor. These results showed that the worm reactors may reduce the waste sludge between 2 and 2.75 times higher than in the blank conditions. The obtained results showed that the worm reactor has a high potential for use in large-scale sludge processing.