bluuebunny/crossref_metadata_2025 huggingface dataset:
Biological and Physicochemical Wastewater Treatment Processes Reduce the Prevalence of Virulent Escherichia coli
Authors: Frigon Dominic , Biswal Basanta Kumar , Mazza Alberto , Masson Luke , Gehr Ronald
Published: November 2012

Effluents discharged from wastewater treatment plants are possible sources of pathogenic bacteria, including
Escherichia coli
, in the freshwater environment, and determining the possible selection of pathogens is important. This study evaluated the impact of activated sludge and physicochemical wastewater treatment processes on the prevalence of potentially virulent
E. coli
. A total of 719
E. coli
isolates collected from four municipal plants in Québec before and after treatment were characterized by using a customized DNA microarray to determine the impact of treatment processes on the frequency of specific pathotypes and virulence genes. The percentages of potentially pathogenic
E. coli
isolates in the plant influents varied between 26 and 51%, and in the effluents, the percentages were 14 to 31%, for a reduction observed at all plants ranging between 14 and 45%. Pathotypes associated with extraintestinal pathogenic
E. coli
(ExPEC) were the most abundant at three of the four plants and represented 24% of all isolates, while intestinal pathogenic
E. coli
pathotypes (IPEC) represented 10% of the isolates. At the plant where ExPEC isolates were not the most abundant, a large number of isolates were classified as both ExPEC and IPEC; overall, 6% of the isolates were classified in both groups, with the majority being from the same plant. The reduction of the proportion of pathogenic
E. coli
could not be explained by the preferential loss of one virulence gene or one type of virulence factor; however, the quinolone resistance gene (
qnrS
) appears to enhance the loss of virulence genes, suggesting a mechanism involving the loss of pathogenicity islands.
Diverse Microhabitats Experienced by Halomonas variabilis on Salt-Secreting Leaves
Authors: Burch Adrien Y. , Finkel Omri M. , Cho Juliana K. , Belkin Shimshon , Lindow Steven E.
Published: November 2012

The leaf surfaces of the salt-excreting treeTamarix aphyllaharbor a wide diversity of halophilic microorganisms, includingHalomonassp., but little is known of the factors that shape community composition in this extreme habitat. We isolated a strain ofHalomonas variabilisfrom the leaf surface ofT. aphyllaand used it to determine the heterogeneity of salt concentrations experienced by bacteria in this environment. This halophilic strain was transformed with aproU::gfpreporter gene fusion, the fluorescence of which was responsive to NaCl concentrations up to 200 g liter−1. These bioreporting cells were applied toT. aphyllaleaves and were subsequently recovered from dew droplets adhering to the leaf surface. Although cells from within a given dew droplet exhibited similar green fluorescent protein fluorescence, the fluorescence intensity varied between droplets and was correlated with the salt concentration measured in each drop. Growth ofH. variabiliswas observed in all droplets, regardless of the salt concentration. However, cells found in desiccated microniches between dew drops were low in abundance and generally dead. Other bacteria recovered fromT. aphylladisplayed higher desiccation tolerance thanH. variabilis, both in culture and on inoculated plants, despite having lower osmotic tolerance. Thus, theTamarixleaf surface can be described as a salty desert with occasional oases where water droplets form under humid conditions. While halotolerant bacteria such asHalomonasgrow in high concentrations of salt in such wet microniches, other organisms are better suited to survive desiccation in sites that are not wetted.
Clinical Specificity of the Enzyme Immunoassay Test for Coccidioidomycosis Varies According to the Reason for Its Performance
Authors: Blair Janis E. , Mendoza Neil , Force Shannon , Chang Yu-Hui H. , Grys Thomas E.
Published: November 2012

The diagnosis of coccidioidomycosis relies heavily on serologic test results in addition to clinical history, physical examination, and radiographic findings. Use of the enzyme immunoassay (EIA) has increased because it is rapidly performed and does not require referral to a reference laboratory, as do complement fixation and immunodiffusion tests. However, interpretation of immunoglobulin M (IgM) reactivity by EIA in the absence of immunoglobulin G (IgG) reactivity has been problematic. We conducted a retrospective medical record review of all patients with such IgM reactivity at our institution to identify situations where the finding was more likely to be clinically specific for coccidioidal infection. From 1 January 2004 through 31 December 2008, a total of 1,117 patients had positive EIA coccidioidal serology or EIA IgM-only reactivity; of these, 102 patients (9%) had EIA IgM-only reactivity. Among the 102 patients with EIA IgM-only reactivity, 60 were tested to evaluate symptomatic illness, 13 for follow-up of previously abnormal serology, and 29 for screening purposes. Of the 102 patients, 80 (78%) had positive serologic findings by other methods or had positive culture or histology. Fifty-four (90%) of the 60 patients whose serology was performed to evaluate symptomatic illness had coccidioidal infection, whereas 13 (45%) of 29 patients whose serology was performed for screening purposes had coccidioidal infection. Of the 102 patients with isolated IgM reactivity by EIA, 12 later seroconverted to IgG and IgM reactivity. The use of EIA for screening in 29 asymptomatic persons was associated with unconfirmable results in 13 (45%). Although the majority of patients in our study with isolated IgM reactivity by EIA had probable or confirmed coccidioidomycosis, this result must be interpreted with caution for asymptomatic patients.
Intracellular Trafficking and Glycobiology of TbPDI2, a Stage-Specific Protein Disulfide Isomerase in Trypanosoma brucei
Authors: Schwartz Kevin J. , Peck Ronald F. , Bangs James D.
Published: November 2012

Trypanosoma brucei
protein disulfide isomerase 2 (TbPDI2) is a bloodstream stage-specific lumenal endoplasmic reticulum (ER) glycoprotein. ER localization is dependent on the TbPDI2 C-terminal tetrapeptide (KQDL) and is mediated by TbERD2, an orthologue of the yeast ER retrieval receptor. Consistent with this function, TbERD2 localizes prominently to ER exit sites, and RNA interference (RNAi) knockdown results in specific secretion of a surrogate ER retention reporter, BiPN:KQDL. TbPDI2 is highly N-glycosylated and is reactive with tomato lectin, suggesting the presence of poly-
N
-acetyllactosamine modifications, which are common on lyso/endosomal proteins in trypanosomes but are inconsistent with ER localization. However, TbPDI2 is reactive with tomato lectin immediately following biosynthesis—far too rapidly for transport to the Golgi compartment, the site of poly-
N
-acetyllactosamine addition. TbPDI2 also fails to react with
Erythrina cristagalli
lectin, confirming the absence of terminal
N
-acetyllactosamine units. We propose that tomato lectin binds the Manβ1-4GlcNAcβ1-4GlcNAc trisaccharide core of paucimannose glycans on both newly synthesized and mature TbPDI2. Consistent with this proposal, α-mannosidase treatment renders oligomannose
N
-glycans on the
T. brucei
cathepsin L orthologue TbCatL reactive with tomato lectin. These findings resolve contradictory evidence on the location and glycobiology of TbPDI2 and provide a cautionary note on the use of tomato lectin as a poly-
N
-acetyllactosamine-specific reagent.
Promoting physical activity in sedentary elderly Malays with type 2 diabetes: a protocol for randomised controlled trial
Authors: Sazlina Shariff-Ghazali , Browning Colette Joy , Yasin Shajahan
Published: November 2012

Introduction

Like many countries Malaysia is facing an increase in the number of people with type 2 diabetes mellitus diabetes (T2DM) and modifiable lifestyle factors such as sedentary behaviour are important drivers of this increase. The level of physical activity is low among elderly Malay people. In Malaysia, strategies to promote physical activity in elderly Malay people with T2DM are not well documented in the research literature. This paper discusses an intervention to increase physical activity in elderly Malay people with T2DM. The aim of our study was to evaluate the effectiveness of personalised feedback alone and in combination with peer support in promoting and maintaining physical activity in comparison with usual care.

Methods and analysis

A three-arm randomised controlled trial will be conducted among sedentary Malay adults aged 60 years and above with T2DM attending an urban primary healthcare clinic in Malaysia. The participants will be randomised into three groups for a 12-week intervention with a follow-up at 24 and 36 weeks to assess adherence. The primary outcome of this study is pedometer-determined physical activity. Glycaemic and blood pressure control, body composition, cardiorespiratory fitness, balance, lipid profile, health-related quality of life, psychological well-being, social support and self-efficacy for exercise are the secondary measures. Linear mixed models will be used to determine the effect of the intervention over time and between groups.

Ethical and dissemination

The Monash University Human Research Ethics Committee and the Malaysian Ministry of Health's Medical Research Ethics Committee approved this protocol. The findings of this study will be presented at international conferences and published in peer-reviewed journals.

Trial registration

This study protocol has been registered with the Malaysian National Medical Research Registry and with the Current Controlled Trial Ltd (http://www.controlled-trials.com/ISRCTN71447000/).
Participation of natural killer cells in the pathogenesis of bile duct lesions in biliary atresia
Authors: Okamura Atsushi , Harada Kenichi , Nio Masaki , Nakanuma Yasuni
Published: November 2012

Aims

Immunological disturbances including innate immunity after a suspected viral infection are considered important to the pathogenesis of bile duct lesions in cases of biliary atresia (BA). In this study, we tried to evaluate whether natural killer (NK) cells and CX3CL1 (Fractalkine) and its receptor (CX3CR1) are involved in the bile duct injury.

Methods

Using the section of BA (22 cases) and controls, immunohistochemistry for CD56, CD16, CD68, CX3CL1 and CX3CR1 was performed. Moreover, using cultured biliary epithelial cells (BECs) and NK cells, the production of CX3CL1 in BECs and the migration of NK cells were evaluated.

Results

It was found that CD56(−)CD16(+)CD68(−) NK cells were increased around the damaged small and large bile ducts in BA and hepatitis C virus-related chronic hepatitis in comparison with other controls. CX3CL1 was strongly expressed on the damaged bile ducts in BA, while this expression was relatively weak or absent in the bile ducts of normal liver. The results suggest the CD56(−)CD16(+) NK cells to be involved in the development of bile duct injuries in BA. These CD16(+) NK cells were positive for CX3CR1, and attracted by CX3CL1 expressed on bile ducts. Further study revealed that stimulation with poly(I:C) (a synthetic analogue of viral dsRNA) increased the expression of CX3CL1 on cultured BECs followed by increased migrational activity of cultured NK cells.

Conclusions

CD56(−)CD16(+) NK cells with reduced NK activity may be involved in the bile duct damage in BA, and CD16(+) NK cells expressing CX3CR1 may be attracted by and interact with bile ducts expressing CX3CL1.
Challenges and proposed solutions for more accurate serological diagnosis of equine infectious anaemia
Authors: Issel C. J. , Scicluna M. T. , Cook S. J. , Cook R. F. , Caprioli A. , Ricci I. , Rosone F. , Craigo J. K. , Montelaro R. C. , Autorino G. L.
Published: November 2012

Serological diagnosis of equine infectious anaemia virus (EIAV) infections has depended mainly on the agar gel immunodiffusion test (AGIDT). This study documents the presence of EIAV genetic sequences in a number of persistently infected horses and mules whose serums were interpreted as negative/equivocal on AGIDT, but positive on more than one ELISA test and in immunoblot tests. Strategies designed to take advantage of the combined strengths of the ELISA and AGIDT are shown effective in a national surveillance program for EIA in Italy where 17 per cent (25/149) of the equids considered to be infected with EIAV on combined/comparative serological data had reactions in the AGIDT that were interpreted as negative or equivocal. These data document the benefits of using a three‐tiered laboratory system for the diagnosis of EIA. Although the ELISA‐first strategy introduces some confusing results, the discovery of up to 20 per cent more cases of EIA makes it compelling. In our opinion, it is better and more defensible to find two samples in 1000 with resolvable but falsely positive ELISA tests for EIA than to release two to three horses in 10,000 with falsely negative test results for EIA (the rates seen in the Italian surveillance presented here).
Tuberculin manufacturing source and breakdown incidence rate of bovine tuberculosis in British cattle, 2005–2009
Authors: Downs S. H. , Clifton‐Hadley R. S. , Upton P. A. , Milne I. C. , Ely E. R. , Gopal R. , Goodchild A. V. , Sayers A. R.
Published: November 2012

The single intradermal comparative cervical tuberculin (SICCT) test is the primary test used for surveillance for bovine tuberculosis (bTB) in cattle in Great Britain (GB). The tuberculin used can, with other factors, influence test accuracy. In this analysis, the detection of infected cattle in GB 2005–2009 was compared between SICCT tests using tuberculins manufacturered by different manufacturers. Higher rates of reactors (adjusted rate 209 vs 186 per 100,000 tests, P = 0.003) and herd bTB incidents (adjusted total breakdown rate 5.1 vs 4.5 per 100 herd‐years at risk, P < 0.001) were detected using tuberculin manufactured at Weybridge compared with Lelystad. However, confirmation of infection in reactors by postmortem evidence was higher with Lelystad tuberculin (adjusted percent 44.1 vs 47.1, P = 0.018). The findings, overall, suggest slightly higher test sensitivity and lower test specificity associated with Weybridge tuberculin compared with Lelystad. Assuming effective adjustment for confounding, the overall impact of tuberculin manufacturing source (2007–2009), was calculated to range somewhere between 315 false positive breakdowns, and 1086 bTB breakdowns missed (624 confirmed) as a result of using Weybridge and Lelystad tuberculin, respectively. However, animals that tested negative to the SICCT were not slaughtered at the time of the tests, so definitive conclusions are not possible.
Particle creation in flat Friedmann–Robertson–Walker (FRW) universe in the framework of f(T) gravity
Authors: Setare M.R. , Houndjo M.J.S.
Published: November 2012

We study particle production in a flat Friedmann–Robertson–Walker universe in the framework of f(T) gravity. An exact power-law solution is obtained by solving the Friedmann equations and assuming that matter is minimally coupled with gravitation. The torsion scalar, T, appears to plays the same role as the curvature (Ricci scalar) in general relativity (GR) and its modified theories, f(R). Particularly, in the phantom phase, we observe that the vacuum state corresponds to a vanishing torsion scalar and particle production becomes important as the torsion scalar diverges. This aspect not only provides the equivalence between teleparallel gravity and GR, but also between their respective modified versions, f(T) and f(R), in the view of massless particle production phenomenon when matter is minimally coupled with gravity. However, when the gravitational and scalar fields are not minimally coupled, it appears that this similarity between the teleparallel gravity and GR may break down, because the torsion scalar no longer has the same time-dependent expression as the Ricci scalar.
Comparative evolutionary genomics of medaka and three-spined sticklebackfabp2aandfabp2bgenes withfabp2of zebrafish
Authors: Parmar Manoj B. , Wright Jonathan M.
Published: November 2012

Here we describe the evolutionary relationship of the duplicated intestinal fatty acid binding protein genes fabp2a and fabp2b from medaka and three-spined stickleback by comparing them to the well-studied fabp2 gene from zebrafish. The duplicated fabp2 genes from medaka and three-spined stickleback consist of four exons separated by three introns, which code for a polypeptide of 132 amino acids. Fabp2a and Fabp2b of medaka and three-spined stickleback share highest sequence identity with zebrafish Fabp2. All Fabp2/FABP2 sequences from vertebrates form a distinct clade in a neighbor-joining phylogenetic tree with a robust 100% bootstrap value, which indicates that the medaka and three-spined stickleback fabp2a and fabp2b are orthologs of zebrafish fabp2. The syntenic genes of fabp2a and fabp2b from medaka and three-spined stickleback were shown to be conserved with the syntenic genes of fabp2/FABP2 from zebrafish and human, evidence that the duplicated fabp2 genes from medaka and three-spined stickleback most likely arose from the teleost-specific whole-genome duplication. The tissue-specific distribution of medaka and three-spined stickleback fabp2a and fabp2b transcripts, and zebrafish fabp2 transcripts, assayed by RT-qPCR suggests the acquisition of new function(s) by the medaka fabp2a, and the distinct evolution of fabp2b compared with fabp2a in the medaka and three-spined stickleback genomes.
Smad7 inhibits autocrine expression of TGF-β2in intestinal epithelial cells in baboon necrotizing enterocolitis
Authors: Namachivayam Kopperuncholan , Blanco Cynthia L. , MohanKumar Krishnan , Jagadeeswaran Ramasamy , Vasquez Margarita , McGill-Vargas Lisa , Garzon Steven A. , Jain Sunil K. , Gill Ravinder K. , Freitag Nancy E. , Weitkamp Jörn-Hendrik , Seidner Steven R. , Maheshwari Akhil
Published: November 2012

Preterm infants may be at risk of necrotizing enterocolitis (NEC) due to deficiency of transforming growth factor-β 2 (TGF-β2) in the developing intestine. We hypothesized that low epithelial TGF-β2expression in preterm intestine and during NEC results from diminished autocrine induction of TGF-β2in these cells. Premature baboons delivered at 67% gestation were treated per current norms for human preterm infants. NEC was diagnosed by clinical and radiological findings. Inflammatory cytokines, TGF-β2, Smad7, Ski, and strawberry notch N (SnoN)/Ski-like oncoprotein (SKIL) was measured using quantitative reverse transcriptase-polymerase chain reaction, immunoblots, and immunohistochemistry. Smad7 effects were examined in transfected IEC6 intestinal epithelial cells in vitro. Findings were validated in archived human tissue samples of NEC. NEC was recorded in seven premature baboons. Consistent with existing human data, premature baboon intestine expressed less TGF-β2than term intestine. TGF-β2expression was regulated in epithelial cells in an autocrine fashion, which was interrupted in the premature intestine and during NEC due to increased expression of Smad7. LPS increased Smad7 binding to the TGF-β2promoter and was associated with dimethylation of the lysine H3K9, a marker of transcriptional silencing, on the nucleosome of TGF-β2. Increased Smad7 expression in preterm intestine was correlated with the deficiency of SnoN/SKIL, a repressor of the Smad7 promoter. Smad7 inhibits autocrine expression of TGF-β2in intestinal epithelial cells in the normal premature intestine and during NEC. Increased Smad7 expression in the developing intestine may be due to a developmental deficiency of the SnoN/SKIL oncoprotein.
Gastrokines: stomach-specific proteins with putative homeostatic and tumor suppressor roles
Authors: Menheniott Trevelyan R. , Kurklu Bayzar , Giraud Andrew S.
Published: November 2012

During the past decade, a new family of stomach-specific proteins has been recognized. Known as “gastrokines” (GKNs), these secreted proteins are products of gastric mucus-producing cell lineages. GKNs are highly conserved in physical structure, and emerging data point to convergent functions in the modulation of gastric mucosal homeostasis and inflammation. While GKNs are highly prevalent in the normal stomach, frequent loss of GKN expression in gastric cancers, coupled with established antiproliferative activity, suggests putative tumor suppressor roles. Conversely, ectopic expression of GKNs in reparative lesions of Crohn's disease alludes to additional activity in epithelial wound healing and/or repair. Modes of action remain unsolved, but the recent demonstration of a GKN2-trefoil factor 1 heterodimer implicates functional interplay with trefoil factors. This review aims to provide a historical account of GKN biology and encapsulate the rapidly accumulating evidence supporting roles in gastric epithelial homeostasis and tumor suppression.
Streptozotocin-induced diabetes differentially affects sympathetic innervation and control of plantar metatarsal and mesenteric arteries in the rat
Authors: Johansen Niloufer J. , Tripovic Diana , Brock James A.
Published: November 2012

In humans neural control of arterial vessels supplying skin in the extremities is particularly vulnerable to the effects of diabetes. Here the streptozotocin (STZ) rat model of type 1 diabetes was used to compare effects on neurovascular function in plantar metatarsal arteries (PMAs), which supply blood to skin of hind paw digits, with those in mesenteric arteries (MAs). Twelve weeks after STZ (60 mg/kg ip), wire myography was used to assess vascular function. In PMAs, lumen dimensions were unchanged but both nerve-evoked contractions and sensitivity to α1 (phenylephrine, methoxamine)- and α2 (clonidine)-adrenoceptor agonists were reduced. The density of perivascular nerve fibers was also reduced by ∼25%. These changes were not observed in PMAs from STZ-treated rats receiving either a low dose of insulin that did not greatly reduce blood glucose levels or a high dose of insulin that markedly reduced blood glucose levels. In MAs from STZ-treated rats, nerve-evoked increases in force did not differ from control but, because lumen dimensions were ∼20% larger, nerve-evoked increases in effective transmural pressure were smaller. Increases in effective transmural pressure produced by phenylephrine or α,β-methylene ATP in MAs from STZ-treated rats were not smaller than control, but the density of perivascular nerve fibers was reduced by ∼10%. In MAs, the increase in vascular dimensions is primarily responsible for reducing effectiveness of nerve-evoked constrictions. By contrast, in PMAs decreases in both the density of perivascular nerve fibers and the reactivity of the vascular muscle appear to explain impairment of neurovascular transmission.
ADAM15 deficiency attenuates pulmonary hyperpermeability and acute lung injury in lipopolysaccharide-treated mice
Authors: Sun Chongxiu , Beard Richard S. , McLean Danielle L. , Rigor Robert R. , Konia Thomas , Wu Mack H. , Yuan Sarah Y.
Published: November 2012

ADAM15 is a disintegrin and metalloprotease recently implicated in cancer and chronic immune disorders. We have recently characterized ADAM15 as a mediator of endothelial barrier dysfunction. Whether this molecule contributes to acute inflammation has not been evaluated. The purpose of this study was to investigate the role of ADAM15 in mediating pulmonary microvascular leakage during acute inflammatory injury. Immunofluorescent staining and Western blotting revealed that the endothelium was the main source of ADAM15 in lung tissue. In a mouse model of acute lung injury induced by lipopolysaccharide (LPS), upregulation of ADAM15 was observed in association with pulmonary edema and neutrophil infiltration. The LPS-induced inflammatory injury, as demonstrated by bronchoalveolar lavage neutrophil count, lung wet-to-dry weight ratio, and myeloperoxidase activity, was significantly attenuated in Adam15 −/− mice. Studies with primary cell culture demonstrated abundant ADAM15 expression in endothelial cells (ECs) of mouse lung but not in neutrophils. Deficiency of ADAM15 in ECs had no obvious effect on basal permeability but significantly attenuated hyperpermeability response to LPS as evidenced by albumin flux assay and measurements of transendothelial electrical resistance, respectively. ADAM15 deficiency also reduced neutrophil chemotactic transmigration across endothelial barriers in the presence or absence of formyl-methionyl-leucyl-phenylalanine (fMLP). Rescue expression of ADAM15 in Adam15 −/− ECs restored neutrophil transendothelial migration. These data indicate that ADAM15 upregulation contributes to inflammatory lung injury by promoting endothelial hyperpermeability and neutrophil transmigration.
Automated measurement of blood flow velocity and direction and hemoglobin oxygen saturation in the rat lung using intravital microscopy
Authors: Hanna Gabi , Fontanella Andrew , Palmer Gregory , Shan Siqing , Radiloff Daniel R. , Zhao Yulin , Irwin David , Hamilton Karyn , Boico Alina , Piantadosi Claude A. , Blueschke Gert , Dewhirst Mark , McMahon Timothy , Schroeder Thies
Published: November 2012

Intravital microscopy of the pulmonary microcirculation in research animals is of great scientific interest for its utility in identifying regional changes in pulmonary microcirculatory blood flow. Although feasibility studies have been reported, the pulmonary window can be further refined into a practical tool for pharmaceutical research and drug development. We have established a method to visualize and quantify dynamic changes in three key features of lung function: microvascular red blood cell velocity, flow direction, and hemoglobin saturation. These physiological parameters were measured in an acute closed-chest pulmonary window, which allows real-time images to be captured by fluorescence and multispectral absorption microscopy; images were subsequently quantified using computerized analysis. We validated the model by quantifying changes in microcirculatory blood flow and hemoglobin saturation in two ways: 1) after changes in inspired oxygen content and 2) after pharmacological reduction of pulmonary blood flow via treatment with the β1 adrenergic receptor blocker metoprolol. This robust and relatively simple system facilitates pulmonary intravital microscopy in laboratory rats for pharmacological and physiological research.
TGF-β-Smad3 signaling in emphysema and pulmonary fibrosis: an epigenetic aberration of normal development?
Authors: Warburton David , Shi Wei , Xu Bing
Published: November 2012

It is well accepted that TGF-β signaling has critical functional roles in lung development, injury, and repair. We showed previously that null mutation of Smad3, a critical node in the TGF-β pathway, protects mice against fibrosis induced by bleomycin. However, more recently we noticed that abnormal alveolarization also occurs in Smad3-deficient mice and that this is followed by progressive emphysema-like alveolar wall destruction mediated by MMP9. We now know that Smad3 cooperates with c-Jun to synergistically regulate a protein deacetylase SIRT1, by binding to an AP-1 site in the SIRT1 promoter. Consistently, Smad3 knockout lung at postnatal day 28 had reduced SIRT1 expression, which in turn resulted in increased histone acetylation at the binding sites of the transcription factors AP-1, NF-κB, and Pea3 on the MMP9 promoter, as well as increased acetylation of NF-κB. Thus, upon TGF-β activation, phosphorylated Smad3 can be translocated into the nucleus with Smad4, whereat Smad3 in turn collaborates with c-Jun to activate SIRT1 transcription. SIRT1 can deacetylate NF-κB at lysine 30, as well as histones adjacent to the transcription factor AP-1, NF-κB, and Pea3 binding sites of the MMP9 promoter, thereby suppressing MMP9 transcription, hence fixing MMP9 in the OFF mode. Conversely, when Smad3 is missing, this regulatory pathway is inactivated so that MMP9 is epigenetically turned ON. We postulate that these developmental epigenetic mechanisms by which Smad3 regulates MMP9 transcription cell autonomously may be important in modulating both emphysema and pulmonary fibrosis and that this could explain why both pathologies can appear within the same lung specimen.
Attitudes to Mathematics in Primary School Children
Authors: Dowker Ann , Bennett Karina , Smith Louise
Published: November 2012

44 Grade 3 children and 45 Grade 5 children from English primary schools were given the British abilities scales basic number skills subtest, and a Mathematics Attitude and Anxiety Questionnaire, using pictorial rating scales to record their Self-rating for maths, Liking for maths, Anxiety about maths, and Unhappiness about poor performance in mathematics. There were few year group differences in attitudes. Boys rated themselves higher than girls, but did not differ significantly in actual performance. Overall, Anxiety was not related to actual performance, but Self-rating was. This relationship between Self-rating and actual performance seemed to develop between Grade 3 and Grade 5. Implications of the findings are discussed.
Precipitation of Phase Using General Diffusion Equation with Comparison to Vitek Diffusion Model in Dissimilar Stainless Steels
Authors: Hsieh Chih-Chun , Wu Weite
Published: November 2012

This study performs a precipitation examination of the phase using the general diffusion equation with comparison to the Vitek model in dissimilar stainless steels during multipass welding. Experimental results demonstrate that the diffusivities (, , and ) of Cr, Ni, and Si are higher in -ferrite than (, , and ) in the phase, and that they facilitate the precipitation of the σ phase in the third pass fusion zone. The Vitek diffusion equation can be modified as follows: .
Fuzzy Support System for Total Hip Arthroplasty Stem by Ultrasonic Intraoperative Measurement
Authors: Yagi Naomi , Hata Yutaka , Shibanuma Nao
Published: November 2012

This paper describes a fuzzy system of stem implantation on total hip arthroplasty by an ultrasonic device. The system can perform automatic and accurate assessment in the surgery. In this system, we employ a single ultrasonic probe whose center frequency is 1,000 Hz. We detect the acoustic signals when knocking the inserted stem with a hammer. We then have a correlation between the degree of tightening and the attenuation time of acoustic signal. That is, the higher tightened degree implies shorter attenuation period. The support system selects the most suitable stem size by fuzzy inference with respect to the attenuation time and its difference time from correct stem to one larger size stem which dynamically adapts to each patient. As a result, we successfully determined the suitable stem in comparison to the results of the practical surgery.
A Long-Term Follow-Up Programme for Maintenance of Motor Function after Stroke: Protocol of the life after Stroke—The LAST Study
Authors: Askim Torunn , Langhammer Birgitta , Ihle-Hansen Hege , Magnussen Jon , Engstad Torgeir , Indredavik Bent
Published: November 2012

Background. There are no evidence-based strategies that have been shown to be superior in maintaining motor function for months to years after the stroke. The LAST study therefore intends to assess the effect of a long-term follow-up program for stroke patients compared to standard care on function, disability and health.Design. This is a prospective, multi-site randomised controlled trial, with blinded assessment 18 months after inclusion. A total of 390 patients will be recruited and randomised to a control group, receiving usual care, or to an intervention group 10 to 16 weeks after onset of stroke. Patients will be stratified according to stroke severity, age above 80, and recruitment site. The intervention group will receive monthly coaching on physical activity by a physiotherapist for 18 consecutive months after inclusion.Outcomes. The primary outcome is motor function (Motor Assessment Scale) 18 months after inclusion. Secondary outcomes are: dependency, balance, endurance, health-related quality of life, fatigue, anxiety and depression, cognitive function, burden on caregivers, and health costs. Adverse events and compliance to the intervention will be registered consecutively during follow-up.
An Unusual Developmental Profile of Salla Disease in a Patient with the SallaFIN Mutation
Authors: Paavola Liisa E. , Remes Anne M. , Sonninen Pirkko H. , Kiviniemi Vesa V. , Korhonen Tapio T. , Majamaa Kari
Published: November 2012

Salla disease (SD) is a disorder caused by defective storage of free sialic acid and results from mutations in theSLC17A5gene. Early developmental delay of motor functions, and later cognitive skills, is typical. We describe a developmental profile of an unusual homozygous patient, who harboured the SallaFIN (p.R39C) mutation gene. The study involved neurological examination, neuropsychological investigation, and brain imaging. The neurocognitive findings were atypical in comparison with other patients with the SallaFIN mutation. Interestingly, there was no deterioration in the patient's neurological condition during adulthood. Her neurocognitive skills were remarkably higher than those of other patients with a conventional phenotype of SD. Our results suggest that the phenotype of SD is broad. Unidentified genetic or environmental variation might explain the unique SD type of this case.
A Qualitative Study of Georgian Youth Who Are on the Street or Institutionalized
Authors: Murray Laura K. , Singh Namrita S. , Surkan Pamela J. , Semrau Katherine , Bass Judy , Bolton Paul
Published: November 2012

Street children, or children who live and/or spend time on the streets, are a vulnerable group of considerable concern to the global public health community. This paper describes the results of two linked qualitative studies conducted with children living or spending time on the street and in orphanages in and around urban areas in the Republic of Georgia between 2005 and 2006. The studies examined perceived causes of children going to the street, as well as indicators of healthy functioning and psychosocial problems among these children. Results on causes indicated a range of “push” factors leading children to the street and “pull” factors that keep children living on the street. Findings also showed a range of internalizing and externalizing mental health symptoms among children on the street and within orphanages. Some differences in responses were found between children living on the street and in institutions. It is important to understand the perspectives of these vulnerable populations to guide decisions on appropriate interventions that address their primary problems.
N‐Formylation of Anilines with Silica Sulfuric Acid under Solvent‐Free Conditions
Authors: Habibi Davood , Rahmani Payam , Akbaripanah Ziba
Published: November 2012

N‐formylation of primary and secondary amines was carried out with formic acid in the presence of silica sulfuric acid under solvent‐free conditions to give the corresponding formamides in excellent yield and short reaction times.
Augmenting Antitumor T-Cell Responses to Mimotope Vaccination by Boosting with Native Tumor Antigens
Authors: Buhrman Jonathan D. , Jordan Kimberly R. , U'Ren Lance , Sprague Jonathan , Kemmler Charles B. , Slansky Jill E.
Published: November 2012

Vaccination with antigens expressed by tumors is one strategy for stimulating enhanced T-cell responses against tumors. However, these peptide vaccines rarely result in efficient expansion of tumor-specific T cells or responses that protect against tumor growth. Mimotopes, or peptide mimics of tumor antigens, elicit increased numbers of T cells that crossreact with the native tumor antigen, resulting in potent antitumor responses. Unfortunately, mimotopes may also elicit cells that do not crossreact or have low affinity for tumor antigen. We previously showed that one such mimotope of the dominant MHC class I tumor antigen of a mouse colon carcinoma cell line stimulates a tumor-specific T-cell clone and elicits antigen-specific cells in vivo, yet protects poorly against tumor growth. We hypothesized that boosting the mimotope vaccine with the native tumor antigen would focus the T-cell response elicited by the mimotope toward high affinity, tumor-specific T cells. We show that priming T cells with the mimotope, followed by a native tumor-antigen boost, improves tumor immunity compared with T cells elicited by the same prime with a mimotope boost. Our data suggest that the improved tumor immunity results from the expansion of mimotope-elicited tumor-specific T cells that have increased avidity for the tumor antigen. The enhanced T cells are phenotypically distinct and enriched for T-cell receptors previously correlated with improved antitumor immunity. These results suggest that incorporation of native antigen into clinical mimotope vaccine regimens may improve the efficacy of antitumor T-cell responses. Cancer Res; 73(1); 74–85. ©2012 AACR.
Alcohol and Acetaldehyde in African Fermented Milk Mursik—A Possible Etiologic Factor for High Incidence of Esophageal Cancer in Western Kenya
Authors: Nieminen Mikko T. , Novak-Frazer Lily , Collins Rebecca , Dawsey Sonja P. , Dawsey Sanford M. , Abnet Christian C. , White Russell E. , Freedman Neal D. , Mwachiro Michael , Bowyer Paul , Salaspuro Mikko , Rautemaa Riina
Published: November 2012

Background: Esophageal cancer is unusually frequent in Western Kenya, despite the low prevalence of classical risk factors such as heavy drinking and tobacco smoking. Among Kenyans consumption of fermented milk is an old tradition. Our hypothesis is that alcohol and acetaldehyde are produced during the fermentation process and that their carcinogenic potential contributes to the high incidence of esophageal cancer.

Methods: Eight samples of mursik milk starter cultures were collected from different Kalenjin families in the Rift Valley province, Western Kenya. A protocol provided by the families was used for milk fermentation. Ethanol and acetaldehyde levels were measured by gas chromatography. The microbial flora in starter cultures was identified by 16S and 18S sequencing.

Results: 7/8 starter cultures produced mutagenic (>100 μmol/L) levels of acetaldehyde and 4/8 starter cultures produced more than 1,000 μmol/L of acetaldehyde. The highest alcohol levels (mean 79.4 mmol/L) were detected in the four fermented milks with highest acetaldehyde production. The mean number of microbial species in the starter cultures was 5 (range 2–8). Yeasts were identified in all starter cultures (mean 1.5 species/milk) but their proportion of the total microbial count varied markedly (mean 35%, range 7%–90%). A combination of yeast and lactobacilli, especially Candida krusei with Lactobacillus kefiri, with the exclusion of other species, seemed to correlate with higher acetaldehyde and ethanol levels.

Conclusions: Significant levels of ethanol and acetaldehyde were produced during mursik fermentation.

Impact: When ingested several times daily the repeated exposure to carcinogenic levels of acetaldehyde may contribute to esophageal carcinogenesis. Cancer Epidemiol Biomarkers Prev; 22(1); 69–75. ©2012 AACR.
A Comparison of Human Papillomavirus Genotype-Specific DNA and E6/E7 mRNA Detection to Identify Anal Precancer among HIV-Infected Men Who Have Sex with Men
Authors: Castle Philip E. , Follansbee Stephen , Borgonovo Sylvia , Tokugawa Diane , Schwartz Lauren M. , Lorey Thomas S. , LaMere Brandon , Gage Julia C. , Fetterman Barbara , Darragh Teresa M. , Rodriguez Ana Cecilia , Wentzensen Nicolas
Published: November 2012

Background: Human papillomavirus (HPV) RNA detection is reportedly more specific for the detection of anogenital precancer than HPV DNA but it is unknown whether this is due to detection of RNA or due to HPV genotype restriction.

Methods: A total of 363 human immunodeficiency virus (HIV)–positive men who have sex with men had two anal cytology samples taken and were evaluated using high-resolution anoscopy and biopsies of visible lesions. Anal specimens were tested for E6/E7 RNA for five carcinogenic HPV genotypes (HPV16, 18, 31, 33, and 45) and tested for the DNA of 13 carcinogenic HPV genotypes.

Results: DNA testing was more likely to be positive than RNA testing (53% vs. 48%; P = 0.02) for the same five HPV genotypes in aggregate. When restricted to five HPV genotypes targeted by the RNA test, the sensitivity to detect anal precancer was the same for DNA and RNA (81%), whereas RNA was more specific than DNA (65% vs. 58%; P = 0.007). In comparison, DNA detection of all 13 carcinogenic HPV genotypes was more sensitive (96% vs. 81%; P = 0.001) but much less specific (65% vs. 33%; P < 0.001) as compared with RNA detection of the five HPV genotypes.

Conclusion: After controlling for HPV genotypes, RNA was only slightly more specific than DNA detection for anal precancer.

Impact: DNA or RNA testing for a subset of the most carcinogenic HPV genotypes may be useful for distinguishing between those HPV-positive men at higher and lower risk of anal precancer and cancer. Cancer Epidemiol Biomarkers Prev; 22(1); 42–9. ©2012 AACR.
Regulation of CXCR4-Mediated Invasion by DARPP-32 in Gastric Cancer Cells
Authors: Zhu Shoumin , Hong Jun , Tripathi Manish K , Sehdev Vikas , Belkhiri Abbes , El-Rifai Wael
Published: November 2012

Although Dopamine and cAMP-regulated phosphoprotein, Mr 32000 (DARPP-32) is overexpressed in two-thirds of gastric cancers, its impact on molecular functions has not been fully characterized. In this study, we examined the role of DARPP-32 in gastric cancer cell invasion. Using matrigel-coated Boyden chamber invasion assay, DARPP-32-overexpressing AGS cells showed a three-fold increase in invasion relative to the vector control (P < 0.01). We also tested the transendothelial cell invasion as a measure of cell aggressiveness using the impedance-based human umbilical vein endothelial cells invasion assay and obtained similar results (P < 0.001). Western blot analysis indicated that overexpression of DARPP-32 mediated an increase in the membrane-type 1 matrix metalloproteinase (MT1-MMP) and CXCR4 protein levels. Consistent with the role of MT1-MMP in cleaving extracellular matrix proteins initiating the activation of soluble MMPs, we detected a robust increase in MMP-2 activity in DARPP-32-overexpressing cells. The knockdown of endogenous DARPP-32 in the MKN-45 cells reversed these signaling events and decreased cell invasive activity. We tested whether the invasive activity mediated by DARPP-32 might involve sustained signaling via CXCR4-dependent activation of the MT1-MMP/MMP-2 pathway. The small-molecule CXCR4 antagonist (AMD3100) and CXCR4-siRNA blocked DARPP-32-induced cell invasion. We further examined our hypothesis that DARPP-32 could interact with CXCR4 and stabilize its levels following stimulation with its ligand, CXCL12. Using reciprocal coimmunoprecipitation and immunofluorescence experiments, we found that DARPP-32 and CXCR4 coexist in the same protein complex. DARPP-32 prolonged the CXCR4 protein half-life and reduced ubiquitination of the CXCR4 protein, following treatment with its ligand, CXCL12. In conclusion, these findings show a novel mechanism by which DARPP-32 promotes cell invasion by regulating CXCR4-mediated activation of the MT1-MMP/MMP-2 pathway. Mol Cancer Res; 11(1); 86–94. ©2012 AACR.
Interleukin-33 Drives a Proinflammatory Endothelial Activation That Selectively Targets Nonquiescent Cells
Authors: Pollheimer Jürgen , Bodin Johanna , Sundnes Olav , Edelmann Reidunn J. , Skånland Sigrid S. , Sponheim Jon , Brox Mari Johanna , Sundlisæter Eirik , Loos Tamara , Vatn Morten , Kasprzycka Monika , Wang Junbai , Küchler Axel M. , Taskén Kjetil , Haraldsen Guttorm , Hol Johanna
Published: November 2012

Objective—

Interleukin (IL)-33 is a nuclear protein that is released from stressed or damaged cells to act as an alarmin. We investigated the effects of IL-33 on endothelial cells, using the prototype IL-1 family member, IL-1β, as a reference.

Methods and Results—

Human umbilical vein endothelial cells were stimulated with IL-33 or IL-1β, showing highly similar phosphorylation of signaling molecules, induction of adhesion molecules, and transcription profiles. However, intradermally injected IL-33 elicited significantly less proinflammatory endothelial activation when compared with IL-1β and led us to observe that quiescent endothelial cells (ppRb
low
p27
high
) were strikingly resistant to IL-33. Accordingly, the IL-33 receptor was preferentially expressed in nonquiescent cells of low-density cultures, corresponding to selective induction of adhesion molecules and chemokines. Multiparameter phosphoflow cytometry confirmed that signaling driven by IL-33 was stronger in nonquiescent cells. Manipulation of nuclear IL-33 expression by siRNA or adenoviral transduction revealed no functional link between nuclear, endogenous IL-33, and exogenous IL-33 responsiveness.

Conclusion—

In contrast to other inflammatory cytokines, IL-33 selectively targets nonquiescent endothelial cells. By this novel concept, quiescent cells may remain nonresponsive to a proinflammatory stimulus that concomitantly triggers a powerful response in cells that have been released from contact inhibition.
Macrophage Activation by Heparanase Is Mediated by TLR-2 and TLR-4 and Associates With Plaque Progression
Authors: Blich Miry , Golan Amnon , Arvatz Gil , Sebbag Anat , Shafat Itay , Sabo Edmond , Cohen-Kaplan Victoria , Petcherski Sirouch , Avniel-Polak Shani , Eitan Amnon , Hammerman Haim , Aronson Doron , Axelman Elena , Ilan Neta , Nussbaum Gabriel , Vlodavsky Israel
Published: November 2012

Objective—

Factors and mechanisms that activate macrophages in atherosclerotic plaques are incompletely understood. We examined the capacity of heparanase to activate macrophages.

Methods and Results—

Highly purified heparanase was added to mouse peritoneal macrophages and macrophage-like J774 cells, and the levels of tumor necrosis factor-α, matrix metalloproteinase-9, interlukin-1, and monocyte chemotactic protein-1 were evaluated by ELISA. Gene expression was determined by RT-PCR. Cells collected from Toll-like receptor-2 and Toll-like receptor-4 knockout mice were evaluated similarly. Heparanase levels in the plasma of patients with acute myocardial infarction, stable angina, and healthy subjects were determined by ELISA. Immunohistochemistry was applied to detect the expression of heparanase in control specimens and specimens of patients with stable angina or acute myocardial infarction. Addition or overexpression of heparanase variants resulted in marked increase in tumor necrosis factor-α, matrix metalloproteinase-9, interlukin-1, and monocyte chemotactic protein-1 levels. Mouse peritoneal macrophages harvested from Toll-like receptor-2 or Toll-like receptor-4 knockout mice were not activated by heparanase. Plasma heparanase level was higher in patients with acute myocardial infarction, compared with patients with stable angina and healthy subjects. Pathologic coronary specimens obtained from vulnerable plaques showed increased heparanase staining compared with specimens of stable plaque and controls.

Conclusion—

Heparanase activates macrophages, resulting in marked induction of cytokine expression associated with plaque progression toward vulnerability.