bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanisms of Human Adenovirus Inactivation by Sunlight and UVC Light as Examined by Quantitative PCR and Quantitative Proteomics
Authors: Bosshard Franziska , Armand Florence , Hamelin Romain , Kohn Tamar 
Published: December 2012

ABSTRACT
Human adenoviruses (HAdV) are important pathogens in both industrialized and developing nations. HAdV has been shown to be relatively resistant to monochromatic UVC light. Polychromatic UVC light, in contrast, is a more effective means of disinfection, presumably due to the involvement of viral proteins in the inactivation mechanism. Solar disinfection of HAdV, finally, is only poorly understood. In this paper, the kinetics and mechanism of HAdV inactivation by UVC light and direct and indirect solar disinfection are elucidated. PCR and mass spectrometry were employed to quantify the extent of genome and protein degradation and to localize the affected regions in the HAdV proteins. For this purpose, we used for the first time an approach involving stable isotope labeling by amino acids in cell culture (SILAC) of a human virus. Inactivation by UVC light and the full sunlight spectrum were found to efficiently inactivate HAdV, whereas UVA-visible light only caused inactivation in the presence of external sensitizers (indirect solar disinfection). Genome damage was significant for UVC but was less important for solar disinfection. In contrast, indirect solar disinfection exhibited extensive protein degradation. In particular, the fiber protein and the amino acids responsible for host binding within the fiber protein were shown to degrade. In addition, the central domain of the penton protein was damaged, which may inhibit interactions with the fiber protein and lead to a disruption of the initial stages of infection. Damage to the hexon protein, however, appeared to affect only regions not directly involved in the infectious cycle.
Heterologous Expression of a Bioactive β-Hexosyltransferase, an Enzyme Producer of Prebiotics, from Sporobolomyces singularis
Authors: Dagher Suzanne F. , Azcarate-Peril M. Andrea , Bruno-Bárcena José M. 
Published: December 2012

ABSTRACTGalacto-oligosaccharides (GOS) are indigestible dietary fibers that are able to reach the lower gastrointestinal tract to be selectively fermented by health-promoting bacteria. In this report, we describe the heterologous expression of an optimized synthetically produced version of the β-hexosyltransferase gene (Bht) fromSporobolomyces singularis. TheBhtgene encodes a glycosyl hydrolase (EC 3.2.1.21) that acts as galactosyltransferase, able to catalyze a one-step conversion of lactose to GOS. Expression of the enzyme inEscherichia coliyielded an inactive insoluble protein, while the methylotrophic yeastPichia pastorisGS115 produced a bioactive β-hexosyltransferase (rBHT). The enzyme exhibited faster kinetics at pHs between 3.5 and 6 and at temperatures between 40 and 50°C. Enzyme stability improved at temperatures lower than 40°C, and glucose was found to be a competitive inhibitor of enzymatic activity.P. pastorissecreted a fraction of the bioactive rBHT into the fermentation broth, while the majority of the enzyme remained associated with the outer membrane. Both the secreted and the membrane-associated forms were able to efficiently convert lactose to GOS. Additionally, resting cells with membrane-bound enzyme converted 90% of the initial lactose into GOS at 68% yield (g/g) (the maximum theoretical is 75%) with no secondary residual (glucose or galactose) products. This is the first report of a bioactive BHT fromS. singularisthat has been heterologously expressed.
Quantification of IncP-1 Plasmid Prevalence in Environmental Samples
Authors: Jechalke Sven , Dealtry Simone , Smalla Kornelia , Heuer Holger 
Published: December 2012

ABSTRACT

To study the role of broad-host-range IncP-1 plasmids in bacterial adaptability to irregular environmental challenges, a quantitative real-time PCR assay was developed that specifically detects the
korB
gene, which is conserved in all IncP-1 plasmids, in environmental samples. IncP-1 plasmid dynamics in a biopurification system for pesticide wastes were analyzed.
Surface Stress Induces a Conserved Cell Wall Stress Response in the Pathogenic Fungus Candida albicans
Authors: Heilmann Clemens J. , Sorgo Alice G. , Mohammadi Sepehr , Sosinska Grazyna J. , de Koster Chris G. , Brul Stanley , de Koning Leo J. , Klis Frans M. 
Published: December 2012

ABSTRACTThe human fungal pathogenCandida albicanscan grow at temperatures of up to 45°C. Here, we show that at 42°C substantially less biomass was formed than at 37°C. The cells also became more sensitive to wall-perturbing compounds, and the wall chitin levels increased, changes that are indicative of wall stress. Quantitative mass spectrometry of the wall proteome using15N metabolically labeled wall proteins as internal standards revealed that at 42°C the levels of the β-glucan transglycosylases Phr1 and Phr2, the predicted chitin transglycosylases Crh11 and Utr2, and the wall maintenance protein Ecm33 increased. Consistent with our previous results for fluconazole stress, this suggests that a wall-remodeling response is mounted to relieve wall stress. Thermal stress as well as different wall and membrane stressors led to an increased phosphorylation of the mitogen-activated protein (MAP) kinase Mkc1, suggesting activation of the cell wall integrity (CWI) pathway. Furthermore, all wall and membrane stresses tested resulted in diminished cell separation. This was accompanied by decreased secretion of the major chitinase Cht3 and the endoglucanase Eng1 into the medium. Consistent with this,cht3cells showed a similar phenotype. When treated with exogenous chitinase, cell clusters both from stressed cells and mutant strains were dispersed, underlining the importance of Cht3 for cell separation. We propose that surface stresses lead to a conserved cell wall remodeling response that is mainly governed by Mkc1 and is characterized by chitin reinforcement of the wall and the expression of remedial wall remodeling enzymes.
Translational Control in Plasmodium and Toxoplasma Parasites
Authors: Zhang Min , Joyce Bradley R. , Sullivan William J. , Nussenzweig Victor 
Published: December 2012

ABSTRACT

The life cycles of apicomplexan parasites such as
Plasmodium
spp. and
Toxoplasma gondii
are complex, consisting of proliferative and latent stages within multiple hosts. Dramatic transformations take place during the cycles, and they demand precise control of gene expression at all levels, including translation. This review focuses on the mechanisms that regulate translational control in
Plasmodium
and
Toxoplasma
, with a particular emphasis on the phosphorylation of the α subunit of eukaryotic translation initiation factor 2 (eIF2α). Phosphorylation of eIF2α (eIF2α∼P) is a conserved mechanism that eukaryotic cells use to repress global protein synthesis while enhancing gene-specific translation of a subset of mRNAs. Elevated levels of eIF2α∼P have been observed during latent stages in both
Toxoplasma
and
Plasmodium
, indicating that translational control plays a role in maintaining dormancy. Parasite-specific eIF2α kinases and phosphatases are also required for proper developmental transitions and adaptation to cellular stresses encountered during the life cycle. Identification of small-molecule inhibitors of apicomplexan eIF2α kinases may selectively interfere with parasite translational control and lead to the development of new therapies to treat malaria and toxoplasmosis.
Fierce Competition between Toxoplasma and Chlamydia for Host Cell Structures in Dually Infected Cells
Authors: Romano Julia D. , de Beaumont Catherine , Carrasco Jose A. , Ehrenman Karen , Bavoil Patrik M. , Coppens Isabelle 
Published: December 2012

ABSTRACT

The prokaryote
Chlamydia trachomatis
and the protozoan
Toxoplasma gondii
, two obligate intracellular pathogens of humans, have evolved a similar
modus operandi
to colonize their host cell and salvage nutrients from organelles. In order to gain fundamental knowledge on the pathogenicity of these microorganisms, we have established a cell culture model whereby single fibroblasts are coinfected by
C. trachomatis
and
T. gondii
. We previously reported that the two pathogens compete for the same nutrient pools in coinfected cells and that
Toxoplasma
holds a significant competitive advantage over
Chlamydia
. Here we have expanded our coinfection studies by examining the respective abilities of
Chlamydia
and
Toxoplasma
to co-opt the host cytoskeleton and recruit organelles. We demonstrate that the two pathogen-containing vacuoles migrate independently to the host perinuclear region and rearrange the host microtubular network around each vacuole. However,
Toxoplasma
outcompetes
Chlamydia
to the host microtubule-organizing center to the detriment of the bacterium, which then shifts to a stress-induced persistent state. Solely in cells preinfected with
Chlamydia
, the centrosomes become associated with the chlamydial inclusion, while the
Toxoplasma
parasitophorous vacuole displays growth defects. Both pathogens fragment the host Golgi apparatus and recruit Golgi elements to retrieve sphingolipids. This study demonstrates that the productive infection by both
Chlamydia
and
Toxoplasma
depends on the capability of each pathogen to successfully adhere to a finely tuned developmental program that aims to remodel the host cell for the pathogen's benefit. In particular, this investigation emphasizes the essentiality of host organelle interception by intravacuolar pathogens to facilitate access to nutrients.
A Repurposing Approach Identifies Off-Patent Drugs with Fungicidal Cryptococcal Activity, a Common Structural Chemotype, and Pharmacological Properties Relevant to the Treatment of Cryptococcosis
Authors: Butts Arielle , DiDone Louis , Koselny Kristy , Baxter Bonnie K. , Chabrier-Rosello Yeissa , Wellington Melanie , Krysan Damian J. 
Published: December 2012

ABSTRACT

New, more accessible therapies for cryptococcosis represent an unmet clinical need of global importance. We took a repurposing approach to identify previously developed drugs with fungicidal activity toward
Cryptococcus neoformans
, using a high-throughput screening assay designed to detect drugs that directly kill fungi. From a set of 1,120 off-patent medications and bioactive molecules, we identified 31 drugs/molecules with fungicidal activity, including 15 drugs for which direct antifungal activity had not previously been reported. A significant portion of the drugs are orally bioavailable and cross the blood-brain barrier, features key to the development of a widely applicable anticryptococcal agent. Structural analysis of this set revealed a common chemotype consisting of a hydrophobic moiety linked to a basic amine, features that are common to drugs that cross the blood-brain barrier and access the phagolysosome, two important niches of
C. neoformans
. Consistent with their fungicidal activity, the set contains eight drugs that are either additive or synergistic in combination with fluconazole. Importantly, we identified two drugs, amiodarone and thioridazine, with activity against intraphagocytic
C. neoformans
. Finally, the set of drugs is also enriched for molecules that inhibit calmodulin, and we have confirmed that seven drugs directly bind
C. neoformans
calmodulin, providing a molecular target that may contribute to the mechanism of antifungal activity. Taken together, these studies provide a foundation for the optimization of the antifungal properties of a set of pharmacologically attractive scaffolds for the development of novel anticryptococcal therapies.
Using Moxibustion in Primary Healthcare to Correct Non-Vertex Presentation: A Multicentre Randomised Controlled Trial
Authors: Vas Jorge , Aranda-Regules José Manuel , Modesto Manuela , Ramos-Monserrat María , Barón Mercedes , Aguilar Inmaculada , Benítez-Parejo Nicolás , Ramírez-Carmona Carmen , Rivas-Ruiz Francisco 
Published: December 2012

Objective To compare the effectiveness of additional moxibustion at point BL67 with moxibustion at a non-specific acupuncture point and with usual care alone to correct non-vertex presentation. Methods This was a multicentre randomised controlled trial in which 406 low-risk pregnant women with a fetus in ultrasound breech presentation, with a gestational age of 33–35 weeks, were assigned to (1) true moxibustion at point BL67 plus usual care; (2) moxibustion at SP1, a non-specific acupuncture point (sham moxibustion) plus usual care; or (3) usual care alone. The primary outcome was cephalic presentation at birth. Women were recruited at health centres in primary healthcare. Results In the true moxibustion group, 58.1% of the full-term presentations were cephalic compared with 43.4% in the sham moxibustion group (RR 1.34, 95% CI 1.05 to 1.70) and 44.8% of those in the usual care group (RR 1.29, 95% CI 1.02 to 1.64). The reduction in RR of the primary outcome in women allocated to the true moxibustion group compared with the usual care group was 29.7% (95% CI 3.1% to 55.2%) and the number needed to treat was 8 (95% CI 4 to 72). There were no severe adverse effects during the treatment. Conclusions Moxibustion at acupuncture point BL67 is effective and safe to correct non-vertex presentation when used between 33 and 35 weeks of gestation. We believe that moxibustion represents a treatment option that should be considered to achieve version of the non-vertex fetus. Trial Registration Current Controlled Trials ISRCTN10634508.
Knee function and knee muscle strength in middle-aged patients with degenerative meniscal tears eligible for arthroscopic partial meniscectomy
Authors: Stensrud Silje , Risberg May Arna , Roos Ewa M 
Published: December 2012

BackgroundFunctional limitations exist postmeniscectomy, but preoperative data are scarce.PurposeTo examine knee function, knee muscle strength and performance in middle-aged patients with degenerative meniscal tears, eligible for arthroscopic partial meniscectomy.Study designCross-sectional study.MethodsEighty-two participants with MRI verified degenerative meniscal tear (35% women, mean age 49 years) answered the Knee injury and Osteoarthritis Outcome Score (KOOS) and were tested for isokinetic knee muscle strength and lower extremity performance (one-leg hop for distance, 6 m timed hop and maximum number of knee-bends in 30 s). Limb Symmetry Index (LSI) was used to express side-to-side differences in per cent using the non-injured leg as the control. An LSI ≥90% was considered normal.ResultsMean scores of the five subscales of the KOOS were from 13 to 36 points lower compared with a population-based reference group and similar to patients prior to anterior cruciate ligament reconstruction. Quadriceps strength and lower-extremity performance were impaired for the injured leg compared with the non-injured leg (p<0.001), with a mean difference of 13% in quadriceps strength and between 8% and 13% in lower-extremity performance. Between 41% and 52% of the participants had abnormal LSI values in quadriceps muscle strength and lower-extremity performance.ConclusionMiddle-aged patients with a symptomatic degenerative meniscal tear experience functional knee problems when eligible for meniscectomy. Included participants reported difficulty with knee pain, symptoms, function and quality of life and quadriceps strength and lower-extremity performance were impaired. Approximately 50% of the study group had clinically relevant impairments in quadriceps strength and lower-extremity performance, defined as >10% differences between the injured and the non-injured leg.
Exposure-adjusted incidence rates and severity of competition injuries in Australian amateur taekwondo athletes: a 2-year prospective study
Authors: Lystad Reidar P , Graham Petra L , Poulos Roslyn G 
Published: December 2012

Background/aimThe main purposes of this study were to determine the injury incidence and severity in Australian amateur taekwondo athletes, and to investigate potential risk factors for injury in competition taekwondo.MethodsData were collected at New South Wales State Championships in 2010 and 2011. Injuries were diagnosed by onsite sports medicine personnel and the actual number of days lost from full participation was used to determine injury severity. Injury incidence rates were calculated per 1000 athlete-exposures (injury incidence rate (IIRAE)) and per 1000 min of exposure (IIRME) and presented with 95% CI.ResultsThe overall IIRAEand IIRMEwere 59.93 (95% CI 51.16 to 69.77) and 16.32 (95% CI 13.93 to 19.00), respectively. Children under 10 years had significantly lower IIRAEcompared with older age groups and black belts had significantly higher IIRAEcompared with yellow belts, however, after accounting for the exposure time it was revealed that 10-year-olds to 14-year-olds and red belts incurred higher IIRME. This study highlights the importance of including IIRs that account for exposure-time. In contrast with previous estimates, the current data indicated that one-third of injuries were moderate to severe. Relative to other body regions the upper limb had a higher proportion of moderate-to-severe injuries, and compared with the lower limb there was a disproportionate number of upper limb injuries resulting in fractures.ConclusionsThe findings suggest that the impact of injury on taekwondo athletes is significant, and should serve as an impetus to stakeholders to develop and implement injury prevention activities within the sport.
Sitting-time and 9-year all-cause mortality in older women
Authors: Pavey Toby G , Peeters GMEE (Geeske) , Brown Wendy J 
Published: December 2012

BackgroundStudies of mid-aged adults provide evidence of a relationship between sitting-time and all-cause mortality, but evidence in older adults is limited. The aim is to examine the relationship between total sitting-time and all-cause mortality in older women.MethodsThe prospective cohort design involved 6656 participants in the Australian Longitudinal Study on Women's Health who were followed for up to 9 years (2002, age 76–81, to 2011, age 85–90). Self-reported total sitting-time was linked to all-cause mortality data from the National Death Index from 2002 to 2011. Cox proportional hazard models were used to examine the relationship between sitting-time and all-cause mortality, with adjustment for potential sociodemographic, behavioural and health confounders.ResultsThere were 2003 (30.1%) deaths during a median follow-up of 6 years. Compared with participants who sat <4 h/day, those who sat 8–11 h/day had a 1.45 times higher risk of death and those who sat ≥11 h/day had a 1.65 times higher risk of death. These risks remained after adding sociodemographic and behavioural covariates, but were attenuated after adjustment for health covariates. A significant interaction (p=0.02) was found between sitting-time and physical activity (PA), with increased mortality risk for prolonged sitting only among participants not meeting PA guidelines (HR for sitting ≥8 h/day: 1.31, 95% CI 1.07 to 1.61); HR for sitting ≥11 h/day: 1.47, CI 1.15 to 1.93).ConclusionsProlonged sitting-time was positively associated with all-cause mortality. Women who reported sitting for more than 8 h/day and did not meet PA guidelines had an increased risk of dying within the next 9 years.
Development and validation of a questionnaire to measure the severity of functional limitations and reduction of sports ability in German-speaking patients with exercise-induced leg pain
Authors: Nauck Tanja , Lohrer Heinz , Padhiar Nat , King John B 
Published: December 2012

BackgroundCurrently, there is no generally agreed measure available to quantify a subject's perceived severity of exercise-induced leg pain symptoms. The aim of this study was to develop and validate a questionnaire that measures the severity of symptoms that impact on function and sports ability in patients with exercise-induced leg pain.MethodsThe exercise-induced leg pain questionnaire for German-speaking patients (EILP-G) was developed in five steps: (1) initial item generation, (2) item reduction, (3) pretesting, (4) expert meeting and (5) validation. The resulting EILP-G was tested for reliability, validity and internal consistency in 20 patients with exercise-induced leg pain, 20 asymptomatic track and field athletes serving as a population at risk and 33 asymptomatic sport students.ResultsThe patient group scored the EILP-G questionnaire significantly lower than both control groups (each p<0.001). Test–retest demonstrates an excellent reliability in all tested groups (Intraclass Correlation Coefficient, ICC=0.861–0.987). Concurrent validity of the EILP-G questionnaire showed a substantial agreement when correlated with the chronic exertional compartment syndrome classification system of Schepsis (r=−0.743; p<0.001). Internal consistency for the EILP-G questionnaire was 0.924.ConclusionsEILP-G questionnaire is a valid and reliable self-administered and disease-related outcome tool to measure the severity of symptoms that impact on function and sports ability in patients with exercise-induced leg pain. It can be recommended as a robust tool for measuring the subjectively perceived severity in German-speaking patients with exercise-induced leg pain.
Are people who do yoga any better at a motor imagery task than those who do not?
Authors: Wallwork Sarah B , Butler David S , Wilson Dianne J , Moseley G Lorimer 
Published: December 2012

BackgroundYoga is a popular recreational activity in Western society and there is an abundance of literature suggesting that yoga may be beneficial for people with a chronic pain disorder. Despite consistently positive results in the literature, the mechanisms of effect are unclear. On the grounds that chronic pain is associated with disruptions of brain-grounded maps of the body, a possible mechanism of yoga is to refine these brain-grounded maps. A left/right body part judgement task is an established way of interrogating these brain-grounded maps of the body.ObjectiveTo determine if people who do regular yoga practice perform better at a left/right judgement task than people who do not.MethodsPreviously collected, cross-sectional data were used. Using a case–control design, participants who reported taking part in regular yoga were selected against age, gender, neck pain and arm pain-matched controls. Participants viewed 40 photographs of a model with their head turned to the left or right, and were asked to judge the direction of neck rotation. They then completed a left/right-hand judgement task.ResultsOf the 1737 participants, 86 of them reported regularly taking part in yoga. From the remaining participants, 86 matched controls were randomly selected from all matched controls. There was no difference between Groups (yoga and no yoga) for either response time (p=0.109) or accuracy (p=0.964). There was a difference between Tasks; people were faster (p<0.001) and more accurate (p=0.001) at making left/right neck rotation judgements than they were at making left/right-hand judgements, regardless of group.ConclusionsPeople who do regular yoga perform no differently in a left/right judgement task than people who do not.