bluuebunny/crossref_metadata_2025_split huggingface dataset:
Optimizing Escherichia coli as a protein expression platform to produce Mycobacterium tuberculosis immunogenic proteins
Authors: Piubelli Luciano , Campa Manuela , Temporini Caterina , Binda Elisa , Mangione Francesca , Amicosante Massimo , Terreni Marco , Marinelli Flavia , Pollegioni Loredano 
Published: November 2013

Abstract

Background
A number of valuable candidates as tuberculosis vaccine have been reported, some of which have already entered clinical trials. The new vaccines, especially subunit vaccines, need multiple administrations in order to maintain adequate life-long immune memory: this demands for high production levels and degree of purity.

Results
In this study, TB10.4, Ag85B and a TB10.4-Ag85B chimeric protein (here-after referred as full) - immunodominant antigens of Mycobacterium tuberculosis - were expressed in Escherichia coli and purified to homogeneity. The rational design of expression constructs and optimization of fermentation and purification conditions allowed a marked increase in solubility and yield of the recombinant antigens. Indeed, scaling up of the process guaranteed mass production of all these three antigens (2.5-25 mg of pure protein/L cultivation broth). Quality of produced soluble proteins was evaluated both by mass spectrometry to assess the purity of final preparations, and by circular dichroism spectroscopy to ascertain the protein conformation. Immunological tests of the different protein products demonstrated that when TB10.4 was fused to Ag85B, the chimeric protein was more immunoreactive than either of the immunogenic protein alone.

Conclusions
We reached the goal of purifying large quantities of soluble antigens effective in generating immunological response against M. tuberculosis by a robust, controlled, scalable and economically feasible production process.
Survey of Plasmodium falciparum multidrug resistance-1 and chloroquine resistance transporter alleles in Haiti
Authors: ElBadry Maha A , Existe Alexandre , Victor Yves S , Memnon Gladys , Fukuda Mark , Dame John B , Yowell Charles A , Okech Bernard A 
Published: November 2013

AbstractBackgroundIn Haiti where chloroquine (CQ) is widely used for malaria treatment, reports of resistance are scarce. However, recent identification of CQ resistance genotypes in one site is suggestive of an emerging problem. Additional studies are needed to evaluate genetic mutations associated with CQ resistance, especially in thePlasmodium falciparummulti-drug resistance-1 gene (pfmdr1) while expanding the already available information onP. falciparumCQ transporter gene (pfcrt) in Haiti.MethodsBlood samples were collected on Whatman filter cards (FTA) from eight clinics spread across Haiti. Following the confirmation ofP. falciparumin the samples, PCR protocols were used to amplify regions ofpfmdr1andpfcrtcodons of interest, (86, 184, 1034, 1042, and 1246) and (72-76), respectively. Sequencing and site-specific restriction enzyme digestions were used to analyse these DNA fragments for the presence of single nucleotide polymorphisms (SNPs) known to confer resistance to anti-malarial drugs.ResultsP. falciparuminfection was confirmed in160 samples by amplifying a segment of theP. falciparum18S small subunit ribosomal RNA gene (pfssurrna). The sequence ofpfmdr1in 54 of these samples was determined between codons 86,184 codons 1034, 1042 and 1246. No sequence differences from that of the NF54 clone 3D7 were found among the 54 samples except at codon 184, where a non-silent mutation was found in all samples predicted to alter the amino acid sequence replacing tyrosine with phenylalanine (Y184F). This altered sequence was also confirmed by restriction enzyme digestion. The sequence ofpfmdr1 at codons 86, 184, 1034 and 1042 encoded the NFSN haplotype. The sequence ofpfcrtcodons 72-76 from 79 samples was determined and found to encode CVMNK, consistent with a CQ sensitive genotype.ConclusionThe presence of the Y184F mutation in pfmdr1ofP. falciparumparasites in Haiti may have implications for resistance to antimalarial drugs. The absence of mutation inpfcrtat codon 76 among 79 isolates tested suggests that sensitivity to CQ in Haiti remains common. Wide-spread screening of thepfmdr1andpfcrtespecially among patients experiencing treatment failure may be a useful tool in early detection of the emergence of antimalarial drug resistance in Haiti.
The relevance of the intrinsic subtype to the clinicopathological features and biomarkers in Japanese breast cancer patients
Authors: Tamaki Masako , Kamio Takako , Kameoka Shingo , Kojimahara Noriko , Nishikawa Toshio 
Published: November 2013

Abstract

Background
Breast cancer is a disease rich in diversity, and it can be categorized into the immunohistochemical intrinsic subtypes : ER/PR + and HER2-, ER/PR + and HER2+, HER2 type, basal-like and unclassified.

Methods
In this study, in addition to the clinicopathological features potentially associated with the intrinsic subtypes, protein expression and genetic mutations of key molecules associated with breast cancer prognosis and treatment sensitivity were analyzed. The distribution of subtypes in the patient population and the differences in marker distribution across the subtypes were investigated.

Results
The immunohistochemical features of 471 consecutive surgical cases of women with primary breast cancer, treated in a single institution, were examined. There were 306 patients who were ER/PR + HER2- (65%); 41 who were ER/PR + HER2+ (8.7%); 59 with HER2 type (12.5%); 37 with basal-like (7.9%); and 28 patients whose breast cancer was unclassified (5.9%). There were no significant differences between the subtypes regarding age, menopausal status, disease stage, lymphatic invasion, blood vessel invasion and lymph node metastasis. Statistically significant differences were found for histological type and grade. Regarding protein expression and genetic mutation, significant differences were found in the distribution within each subtype for six out of 12 molecules investigated.

Conclusions
This study revealed that subtypes differ not only in their clinical pathological profiles, such as histological types and histological grades, but also in molecular expression. The molecular expression patterns observed for each intrinsic subtype may help the selection of an optimal treatment strategy.
Association between community garden participation and fruit and vegetable consumption in rural Missouri
Authors: Barnidge Ellen K , Hipp Pamela R , Estlund Amy , Duggan Kathleen , Barnhart Kathryn J , Brownson Ross C 
Published: November 2013

Abstract

Background
Fruit and vegetable consumption reduces chronic disease risk, yet the majority of Americans consume fewer than recommended. Inadequate access to fruits and vegetables is increasingly recognized as a significant contributor to low consumption of healthy foods. Emerging evidence shows the effectiveness of community gardens in increasing access to, and consumption of, fruits and vegetables.

Methods
Two complementary studies explored the association of community garden participation and fruit and vegetable consumption in rural communities in Missouri. The first was with a convenience sample of participants in a rural community garden intervention who completed self-administered surveys. The second was a population-based survey conducted with a random sample of 1,000 residents in the intervention catchment area.

Results
Participation in a community garden was associated with higher fruit and vegetable consumption. The first study found that individuals who worked in a community garden at least once a week were more likely to report eating fruits and vegetables because of their community garden work (X2 (125) = 7.78, p = .0088). Population-based survey results show that 5% of rural residents reported participating in a community garden. Those who reported community garden participation were more likely to report eating fruits 2 or more times per day and vegetables 3 or more times per day than those who did not report community garden participation, even after adjusting for covariates (Odds Ratio [OR] = 2.76, 95% Confidence Interval [CI] = 1.35 to 5.65).

Conclusion
These complementary studies provide evidence that community gardens are a promising strategy for promoting fruit and vegetable consumption in rural communities.
Brouwer fixed point theorem in "Equation missing"
Authors: Drapeau Samuel , Karliczek Martin , Kupper Michael , Streckfuß Martin 
Published: November 2013

Abstract
The classical Brouwer fixed point theorem states that in

R
d

every continuous function from a convex, compact set on itself has a fixed point. For an arbitrary probability space, let

L
0

=

L
0

(
Ω
,
A
,
P
)

be the set of random variables. We consider

(

L
0

)

d

as an

L
0

-module and show that local, sequentially continuous functions on

L
0

-convex, closed and bounded subsets have a fixed point which is measurable by construction.

MSC:47H10, 13C13, 46A19, 60H25.
Human Papillomavirus and Epstein-Barr virus co-infection in Cervical Carcinoma in Algerian women
Authors: Khenchouche Abdelhalim , Sadouki Nabila , Boudriche Arab , Houali Karim , Graba Abdelaziz , Ooka Tadamasa , Bouguermouh Abdelmadjid 
Published: November 2013

Abstract

Background
Despite the fact that the implication of human papillomavirus (HPV) in the carcinogenesis and prognosis of cervical cancer is well established, the impact of a co-infection with high risk HPV (HR-HPV) and Epstein-Barr virus (EBV) is still not fully understood.

Methods
Fifty eight randomly selected cases of squamous cell carcinomas (SCC) of the uterine cervix, 14 normal cervices specimens, 21 CIN-2/3 and 16 CIN-1 cases were examined for EBV and HPV infections. Detection of HR-HPV specific sequences was carried out by PCR amplification using consensus primers of Manos and by Digene Hybrid Capture. The presence of EBV was revealed by amplifying a 660 bp specific EBV sequence of BALF1. mRNA expression of LMP-1 in one hand and protein levels of BARF-1, LMP-1 and EBNA-1 in the other hand were assessed by RT-PCR and immunoblotting and/or immunohischemistry respectively.

Results
HR-HPV infection was found in patients with SCC (88%), low-grade (75%) and high grade (95%) lesions compared to only 14% of normal cervix cases. However, 69%, 12.5%, 38.1%, and 14% of SCC, CIN-1, CIN-2/3 and normal cervix tissues, respectively, were EBV infected. The highest co-infection (HR-HPV and EBV) was found in squamous cell carcinoma cases (67%). The latter cases showed 27% and 29% expression of EBV BARF-1 and LMP-1 oncogenes respectively.

Conclusion
The high rate of HR-HPV and EBV co-infection in SCC suggests that EBV infection is incriminated in cervical cancer progression. This could be taken into account as bad prognosis in this type of cancer. However, the mode of action in dual infection in cervical oncogenesis needs further investigation.
A device for single leaf labelling with CO2 isotopes to study carbon allocation and partitioning in Arabidopsis thaliana
Authors: Kölling Katharina , Müller Antonia , Flütsch Patrick , Zeeman Samuel C 
Published: November 2013

Abstract

Background
Plant biomass consists primarily of carbohydrates derived from photosynthesis. Monitoring the assimilation of carbon via the Calvin-Benson cycle and its subsequent utilisation is fundamental to understanding plant growth. The use of stable and radioactive carbon isotopes, supplied to plants as CO2, allows the measurement of fluxes through the intermediates of primary photosynthetic metabolism, long-distance transport of sugars in the vasculature, and the synthesis of structural and storage components.

Results
Here we describe the design of a system for supplying isotopically labelled CO2 to single leaves of Arabidopsis thaliana. We demonstrate that the system works well using short pulses of 14CO2 and that it can be used to produce robust qualitative and quantitative data about carbon export from source leaves to the sink tissues, such as the developing leaves and the roots. Time course experiments show the dynamics of carbon partitioning between storage as starch, local production of biomass, and export of carbon to sink tissues.

Conclusion
This isotope labelling method is relatively simple to establish and inexpensive to perform. Our use of 14CO2 helps establish the temporal and spatial allocation of assimilated carbon during plant growth, delivering data complementary to those obtained in recent studies using 13CO2 and MS-based metabolomics techniques. However, we emphasise that this labelling device could also be used effectively in combination with 13CO2 and MS-based techniques.
Doppler ultrasound evaluation in preeclampsia
Authors: Lopez-Mendez Maria A , Martinez-Gaytan Victoria , Cortes-Flores Raul , Ramos-Gonzalez Rene M , Ochoa-Torres Mauro A , Garza-Veloz Idalia , Martinez-Acuña Monica I , Badillo-Almaraz Jose I , Martinez-Fierro Margarita L 
Published: November 2013

Abstract

Background
Worldwide preeclampsia (PE) is the leading cause of maternal death and affects 5 to 8% of pregnant women. PE is characterized by elevated blood pressure and proteinuria. Doppler Ultrasound (US) evaluation has been considered a useful method for prediction of PE; however, there is no complete data about the most frequently altered US parameters in the pathology. The aim of this study was to evaluate the uterine, umbilical, and the middle cerebral arteries using Doppler US parameters [resistance index (RI), pulsatility index (PI), notch (N), systolic peak (SP) and their combinations] in pregnant women, in order to make a global evaluation of hemodynamic repercussion caused by the established PE.

Results
A total of 102 pregnant Mexican women (65 PE women and 37 normotensive women) were recruited in a cases and controls study. Blood velocity waveforms from uterine, umbilical, and middle cerebral arteries, in pregnancies from 24 to 37 weeks of gestation were recorded by trans-abdominal examination with a Toshiba Ultrasound Power Vision 6000 SSA-370A, with a 3.5 MHz convex transducer. Abnormal general Doppler US profile showed a positive association with PE [odds ratio (OR) = 2.93, 95% confidence interval (CI) = 1.2 - 7.3, P = 0.021)], and a specificity and predictive positive value of 89.2% and 88.6%, respectively. Other parameters like N presence, RI and PI of umbilical artery, as well as the PI of middle cerebral artery, showed differences between groups (P values < 0.05).

Conclusion
General Doppler US result, as well as N from uterine vessel, RI from umbilical artery, and PI from umbilical and middle cerebral arteries in their individual form, may be considered as tools to determine hemodynamic repercussion caused by PE.
Parasite epigenetics and immune evasion: lessons from budding yeast
Authors: Wyse Brandon A , Oshidari Roxanne , Jeffery Daniel CB , Yankulov Krassimir Y 
Published: November 2013

AbstractThe remarkable ability of many parasites to evade host immunity is the key to their success and pervasiveness. The immune evasion is directly linked to the silencing of the members of extended families of genes that encode for major parasite antigens. At any time only one of these genes is active. Infrequent switches to other members of the gene family help the parasites elude the immune system and cause prolonged maladies. For most pathogens, the detailed mechanisms of gene silencing and switching are poorly understood. On the other hand, studies in the budding yeastSaccharomyces cerevisiaehave revealed similar mechanisms of gene repression and switching and have provided significant insights into the molecular basis of these phenomena. This information is becoming increasingly relevant to the genetics of the parasites. Here we summarize recent advances in parasite epigenetics and emphasize the similarities betweenS. cerevisiaeand pathogens such asPlasmodium,Trypanosoma,Candida, andPneumocystis. We also outline current challenges in the control and the treatment of the diseases caused by these parasites and link them to epigenetics and the wealth of knowledge acquired from budding yeast.
Will vaccination against rotavirus infection with RIX4414 be cost-saving in Germany?
Authors: Knoll Stefanie , Mair Christoph , Benter Ursula , Vouk Katja , Standaert Baudouin 
Published: November 2013

AbstractBackgroundRotavirus gastroenteritis (RVGE) is a frequent disease in young children. The recommended German paediatric immunisation schedule does not currently include rotavirus vaccination. A lack of economic data on the impact of routine vaccination is stated as one of the reasons. As a result, the current coverage rate is low, around 26%. This study investigated whether rotavirus vaccination using the two-dose rotavirus vaccine RIX4414 (Rotarix®, GlaxoSmithKline Vaccines) would be a cost-saving intervention from the perspective of the statutory health insurance (SHI) in Germany.ObjectiveThe objective of the study was to analyse health outcomes (number of RVGE cases and hospitalisations prevented) and the associated cost to the SHI when comparing 100% rotavirus vaccination with no vaccination in Germany.MethodsA Markov cohort model simulated the number of RVGE events and related costs in a German birth cohort over the first 60 months of life with current disease management. The model compared an unvaccinated cohort with a fully vaccinated cohort. Vaccine efficacy data from international clinical trials were combined with German-specific epidemiological and cost data. Results were tested using extensive sensitivity analyses.ResultsFull vaccination of a birth cohort against rotavirus disease would be expected to prevent 82% of RVGE cases, reducing RVGE frequency from 28 to 5 events per 100 children in the birth cohort up to age 5 years. The estimated cost reduction with vaccination for that period is predicted to be €9.2 million with 100% coverage (€6.9 million with 75% coverage), mainly due to reductions in SHI reimbursement for productivity losses, hospital stays and visits to office-based physicians due to the vaccine’s efficacy against severe disease.ConclusionsRoutine rotavirus vaccination in Germany would reduce the number of hospitalised and outpatient cases. The associated investment could be fully offset by costs avoided in hospital stays, physician visits and SHI reimbursement of productivity losses. Sensitivity analysis indicated that vaccination would be cost-saving in 95% of simulations. Incremental cost was observed only under extreme conditions, especially when the time spent at home due to rotavirus disease was low or when vaccine efficacy against severe disease was heavily decreased.
Experimental measurement of surface strains and local lattice rotations combined with 3D microstructure reconstruction from deformed polycrystalline ensembles at the micro-scale
Authors: Shade Paul A , Groeber Michael A , Schuren Jay C , Uchic Michael D 
Published: November 2013

AbstractThis article describes a new approach to characterize the deformation response of polycrystalline metals using a combination of novel micro-scale experimental methodologies. An in-situ scanning electron microscope (SEM)-based tension testing system was used to deform micro-scale polycrystalline samples to modest and moderate plastic strains. These tests included measurement of the local displacement field with nm-scale resolution at the sample surface. After testing, focused ion beam serial sectioning experiments that incorporated electron backscatter diffraction mapping were performed to characterize both the internal 3D grain structure and local lattice rotations that developed within the deformed micro-scale test samples. This combination of experiments enables the local surface displacements and internal lattice rotations to be directly correlated with the underlying 3D polycrystalline microstructure, and such information can be used to validate and guide further development of modeling and simulation methods that predict the local plastic deformation response of polycrystalline ensembles.
The inclusion of N-terminal pro-brain natriuretic peptide in a sensitive screening strategy for systemic sclerosis-related pulmonary arterial hypertension: a cohort study
Authors: Thakkar Vivek , Stevens Wendy , Prior David , Youssef Peter , Liew Danny , Gabbay Eli , Roddy Janet , Walker Jennifer G , Zochling Jane , Sahhar Joanne , Nash Peter , Lester Susan , Rischmueller Maureen , Proudman Susanna M , Nikpour Mandana 
Published: November 2013

Abstract
Introduction
Pulmonary arterial hypertension (PAH) is a major cause of mortality in systemic sclerosis (SSc). Screening guidelines for PAH recommend multiple investigations, including annual echocardiography, which together have low specificity and may not be cost-effective. We sought to evaluate the predictive accuracy of serum N-terminal pro-brain natriuretic peptide (NT-proBNP) in combination with pulmonary function tests (PFT) (‘proposed’ algorithm) in a screening algorithm for SSc-PAH.

Methods
We evaluated our proposed algorithm (PFT with NT-proBNP) on 49 consecutive SSc patients with suspected pulmonary hypertension undergoing right heart catherisation (RHC). The predictive accuracy of the proposed algorithm was compared with existing screening recommendations, and is presented as sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV).

Results
Overall, 27 patients were found to have pulmonary hypertension (PH) at RHC, while 22 had no PH. The sensitivity, specificity, PPV and NPV of the proposed algorithm for PAH was 94.1%, 54.5%, 61.5% and 92.3%, respectively; current European Society of Cardiology (ESC)/European Respiratory Society (ERS) guidelines achieved a sensitivity, specificity, PPV and NPV of 94.1%, 31.8%, 51.6% and 87.5%, respectively. In an alternate case scenario analysis, estimating a PAH prevalence of 10%, the proposed algorithm achieved a sensitivity, specificity, PPV and NPV for PAH of 94.1%, 54.5%, 18.7% and 98.8%, respectively.

Conclusions
The combination of NT-proBNP with PFT is a sensitive, yet simple and non-invasive, screening strategy for SSc-PAH. Patients with a positive screening result can be referred for echocardiography, and further confirmatory testing for PAH. In this way, it may be possible to shift the burden of routine screening away from echocardiography. The findings of this study should be confirmed in larger studies.
SerpinB1 regulates homeostatic expansion of IL-17+ γδ and CD4+ Th17 cells
Authors: Zhao Picheng , Hou Lifei , Farley Kalamo , Sundrud Mark S , Remold-O'Donnell Eileen 
Published: November 2013

ABSTRACTSerpinB1 is an endogenous inhibitor of serine proteases recognized for its anti-inflammatory and host-protective properties. Although loss of serpinB1 in mice does not result in gross immune deregulation, serpinb1a−/− mice display increased mortality and inflammation-associated morbidity upon challenge with influenza virus. Here, we show that IL-17A+ γδ and CD4+ Th17 cells are already expanded in the lungs of serpinb1a−/− mice at steady-state. Both γδ and αβ+ CD4+ CCR6+ T cells isolated from the lungs of naive serpinb1a−/− mice displayed a skewed transcriptional profile relative to WT cells, including increased Th17 signature transcripts [Il17a, l17f, and Rorc (RORγt)] and decreased Th1 signature transcripts [Ifng, Cxcr3, and Tbx21 (T-bet)] in γδ T cells. In addition to the lung, IL-17A+ γδ and CD4+ Th17 cells were increased in the spleen of naive serpinb1a−/− mice, despite normal αβ and γδ T cell development in the thymus. Within the γδ T cell compartment, loss of serpinb1a prompted selective expansion of Vγ4+ and Vγ6/Vδ1+ cells, which also displayed elevated expression of the proliferating cell nuclear antigen, Ki-67, and IL-17A. Given that serpinb1a is preferentially expressed in WT IL-17A+ γδ and CD4+ Th17 cell subsets vis-à-vis other T cell lineages, our findings reveal a novel function of serpinB1 in limiting untoward expansion of lymphocytes with a Th17 phenotype.