bluuebunny/crossref_metadata_2025 huggingface dataset:
The SOAR (Stroke Subtype, Oxford Community Stroke Project Classification, Age, Prestroke Modified Rankin) Score Strongly Predicts Early Outcomes in Acute Stroke
Authors: Myint Phyo Kyaw , Clark Allan B. , Kwok Chun Shing , Davis John , Durairaj Ramesh , Dixit Anand K. , Sharma Anil K. , Ford Gary A. , Potter John F.
Published: July 2013

Background

Previous prognostic scoring systems in predicting stroke mortality are complex, require multiple measures that vary with time and failed to produce a simple scoring system.

Aims/Hypothesis

The study aims to derive and internally validate a stroke prognostic scoring system to predict early mortality and hospital length of stay.

Methods

Data from a UK multicenter stroke register were examined (1997–2010). Using a prior hypothesis based on our and others observations, we selected five patient-related factors (age, gender, stroke subtype, clinical classification, and prestroke disability) as candidate prognostic indicators. An 8-point score was derived based on multiple logistic regression model using four out of five variables. Performance of the model was assessed by plotting the estimated probability of in-hospital death against the actual probability by testing for overfitting (calibration) and area under the curve methods (discrimination).

Results

The total sample consisted of 12 355 acute stroke patients (ischemic stroke 91·0%). The score predicted both in-patient and seven-day mortality. The crude in-patient mortality were 1·57%, 4·02%, 10·65%, 21·41%, 46·60%, 62·72%, and 75·81% for those who scored 0, 1, 2, 3, 4, 5, and 6, respectively. The calibration of the model revealed no evidence of overfitting (estimated overfitting 0·001). The area under the curve values for both in-hospital and seven-day mortality were 0·79. The score predicted length of stay with a higher score was associated with longer median length of stay in those discharged alive and shorter median length of stay in those who died (P for both <0·001).

Conclusions

A simple 8-point clinical score is highly predictive of acute stroke mortality and length of hospital stay. It could be used as prognostic tool in service planning and also to risk-stratify patients to use these outcomes as markers of stroke care quality across institutions.
Assessment of alcohol histories obtained from patients with liver disease: opportunities to improve early intervention
Authors: Fagan K. J. , Irvine K. M. , Kumar S. , Bates A. , Horsfall L. U. , Feeney G. F. , Powell E. E.
Published: July 2013

Background

Alcohol is an important primary and comorbid cause of liver injury in patients referred for investigation and management of liver disease. Early assessment and documentation of alcohol consumption is therefore essential, and recommended in both general practice and hospital settings.

Aims

To determine the extent and accuracy of documentation of alcohol consumption in patients referred for evaluation of liver disease.

Methods

Patients were interviewed using a structured questionnaire. The medical records of all patients interviewed were reviewed to obtain information from the referral letter and the hepatology consultations.

Results

Eighty‐three patients were surveyed. Only 14 referrals had an informative alcohol history, despite 27 patients admitting risky alcohol consumption at the initial hepatology consultation. Ninety per cent of initial consultations had an informative alcohol history documented, whereas only 56% of patients attending a follow‐up appointment had informative documentation. Assessment of alcohol consumption was comparable between the hepatology consultation and the structured questionnaire, but four subjects had substantially different alcohol histories. Alcohol Use Disorders Identification Test identified all patients reporting harmful alcohol consumption on the questionnaire.

Conclusions

Hazardous alcohol use is prevalent in subjects attending hepatology clinics, but informative alcohol histories, which are crucial to patient management, are rarely documented in referrals. Screening tools improve documentation and accuracy of alcohol histories, and their use by general practitioners and hospital clinicians would improve detection rates of hazardous drinking and allow earlier intervention. Systematic use of screening tools in hepatology clinics will provide opportunities for education and reinforce recommendations to reduce hazardous or harmful alcohol consumption.
Developing a national database for metastatic colorectal cancer management: perspectives and challenges
Authors: Field K. , Wong H.‐L. , Shapiro J. , Kosmider S. , Tie J. , Bae S. , Yip D. , McKendrick J. , Nott L. , Desai J. , Harold M. , Lipton L. , Stefanou G. , Lim L. , Parente P. , Gibbs P.
Published: July 2013

Background

The changing treatment landscape for metastatic colorectal cancer creates multiple potential treatment strategies. An Australian‐centric database capturing comprehensive information across a range of treatment locations would create a valuable resource enabling multiple important research questions to be addressed.

Aims

To establish a collection of a consensus dataset capturing treatment and outcomes at multiple public and private hospitals across Australia.

Methods

An electronic database was developed by a panel of clinicians, to capture an agreed dataset for patients with newly diagnosed metastatic colorectal cancer. Of particular interest were clinician decision‐making, the impact of comorbidities and the frequency of major adverse events.

Results

Since July 2009, data collection has been established at six public and eight private hospitals across three Australian states and territories. Successful linkage and analysis, with support from BioGrid Australia, of selected data on the initial 864 patients demonstrates that data can be captured from diverse sites, including public and private practice, that multiple factors impact on treatment delivered and outcomes achieved and that comprehensive data on rare but important adverse events can be captured. As a clinical research tool, the project has been highly successful, generating multiple presentations at national and international conferences related to a diverse range of research questions.

Conclusions

Multistate, project‐specific data collection involving large numbers of patients is achievable. Providing invaluable insight into the routine clinical management of metastatic colorectal cancer in the era of targeted therapies, this also creates a significant resource for research, including many questions not being addressed by clinical trials.
Effectiveness of an acellular synthetic matrix in the treatment of hard‐to‐heal leg ulcers
Authors: Harding Keith , Aldons Pat , Edwards Helen , Stacey Michael , Finlayson Kathleen , Gibb Michelle , Jenkins Liz , Shooter Gary , Lonkhuyzen Derek Van , Lynam Emily , Heinrichs Eva‐Lisa , Upton Zee
Published: July 2013

Hard‐to‐heal leg ulcers are a major cause of morbidity in the elderly population. Despite improvements in wound care, some wounds will not heal and they present a significant challenge for patients and health care providers. A multi‐centre cohort study was conducted to evaluate the effectiveness and safety of a synthetic, extracellular matrix protein as an adjunct to standard care in the treatment of hard‐to‐heal venous or mixed leg ulcers. Primary effectiveness criteria were (i) reduction in wound size evaluated by percentage change in wound area and (ii) healing assessed by number of patients healed by end of the 12 week study. Pain reduction was assessed as a secondary effectiveness criteria using VAS. A total of 45 patients completed the study and no difference was observed between cohorts for treatment frequency. Healing was achieved in 35·6% and wound size decreased in 93·3% of patients. Median wound area percentage reduction was 70·8%. Over 50% of patients reported pain on first visit and 87·0% of these reported no pain at the end of the study. Median time to first reporting of no pain was 14 days after treatment initiation. The authors consider the extracellular synthetic matrix protein an effective and safe adjunct to standard care in the treatment of hard‐to‐heal leg ulcers.
Degradation of MSCRAMM target macromolecules in VLU slough by Lucilia sericata chymotrypsin 1 (ISP) persists in the presence of tissue gelatinase activity
Authors: Pritchard David I , Brown Alan P
Published: July 2013

Venous leg ulcer slough is unpleasant to the patient and difficult to manage clinically. It harbours infection, also preventing wound management materials and dressings from supporting the underlying viable tissues. In other words, slough has significant nuisance value in the tissue viability clinic. In this study, we have sought to increase our knowledge of slough by building upon a previous but limited analysis of this necrotic tissue. In particular, slough has been probed using Western blotting for the presence of proteins with the capacity to engage microbial surface components recognising adhesive matrix macromolecules. Although the samples were difficult to resolve, we detected fibrinogen, fibronectin, IgG, collagen, human serum albumin and matrix metalloproteinase‐9. Furthermore, the effect of a maggot‐derived debridement enzyme, chymotrypsin 1 on macromolecules in slough was confirmed across seven patient samples. The effect of chymotrypsin 1 on slough confirms our thesis that this potential debridement enzyme could be effective in removing slough along with its associated bacteria, given its observed resistance to intrinsic gelatinase activity. In summary, we believe that the data provide scientists and clinicians with further insights into the potential molecular interactions between bacteria, wound tissue and Lucilia sericata in a clinically problematic yet scientifically interesting wound ecosystem.
“Let Every Christian Denomination Cheerfully Unite”: The Origins of Presbyterian Interdenominationalism
Authors: Taylor William Harrison
Published: July 2013

The history of religion during the eighteenth century is, fortunately, a well‐developed and researched field. Despite the strides taken, however, little has been written on denominational attempts at Christian unity. Historians have instead focused on the multitude of conflicts, both social and religious, that marked the period and preoccupied churchgoers. Although this perspective is indispensable for any understanding of the eighteenth century, it is incomplete. The current portrayal of the late colonial religious scene as one of violently opposed denominations presents the well‐known instances of denominational unity, such as the bishopric crisis, the constitutional crisis, and the War for Independence, as products of political or temporal motivations. Overlooked are the religiously motivated attempts between churches to cooperate, such as the interdenominational journey begun by the Presbyterian Church during the French and Indian War. By examining the Presbyterian struggle to establish a stronger spiritual bond between Christian denominations, it sheds new light which calls into question the current understanding of church participation in the pivotal events of the eighteenth century. Harkened by a divine punishment, Presbyterian interdenominationalism reveals not only that ecclesiastical harmony was pursued in an era defined by conflict, but that these unions could also be motivated by religious rather than solely political ideology.
Genetic background, and not ontogenetic effects, affects avian seasonal timing of reproduction
Authors: Gienapp P. , van Noordwijk A. J. , Visser M. E.
Published: July 2013

Avian seasonal timing is a life‐history trait with important fitness consequences and which is currently under directional selection due to climate change. To predict micro‐evolution in this trait, it is crucial to properly estimate its heritability. Heritabilities are often estimated from pedigreed wild populations. As these are observational data, it leaves the possibility that the resemblance between related individuals is not due to shared genes but to ontogenetic effects; when the environment for the offspring provided by early laying pairs differs from that by late pairs and the laying dates of these offspring when they reproduce themselves is affected by this environment, this may lead to inflated heritability estimates. Using simulation studies, we first tested whether and how much such an early environmental effect can inflate heritability estimates from animal models, and we showed that pedigree structure determines by how much early environmental effects inflate heritability estimates. We then used data from a wild population of great tits (Parus major) to compare laying dates of females born early in the season in first broods and from sisters born much later, in second broods. These birds are raised under very different environmental conditions but have the same genetic background. The laying dates of first and second brood offspring do not differ when they reproduce themselves, clearly showing that ontogenetic effects are very small and hence, family resemblance in timing is due to genes. This finding is essential for the interpretation of the heritabilities reported from wild populations and for predicting micro‐evolution in response to climate change.
Flyingfish feeding ecology in the eastern Pacific: prey partitioning within a speciose epipelagic community
Authors: Van Noord J. E. , Lewallen E. A. , Pitman R. L.
Published: July 2013

To test the hypothesis that prey partitioning contributes to community stability in flyingfish, the gut contents of 359 flyingfish specimens (representing five genera and eight species within Exocoetidae and Hemiramphidae) were collected at 50 dip‐net stations during hour‐long night‐time fishing in oceanic waters of the eastern tropical Pacific Ocean between August and November 2007. Analyses using multidimensional scaling, and analysis of similarity revealed significant dietary differences among species, and similarity percentages tests helped identify the specific prey taxa responsible for these differences. Six species specialized on copepods (58·3–96·9% by number), but targeted different families. Specifically, the barbel flyingfish Exocoetus monocirrhus (n = 205) focused on euchaetids (51·6%), the banded flyingfish Hirundichthys marginatus (n = 24) fed on pontellids (21·8%), while the tropical two‐wing flyingfish Exocoetus volitans (n = 11) and the bigwing halfbeak Oxyporhamphus micropterus (n = 34) ingested calanoids (54·6 and 17·0%). In contrast, the whitetip flyingfish Cheilopogon xenopterus (n = 73) and the mirrorwing flyingfish Hirundichthys speculiger (n = 4) had generalized diets comprising similar proportions of amphipod, copepod, mollusc and larval fish prey. Distinct differences in mean fullness, highly digested material, per cent empty guts and mean numbers of prey per gut were also synthesized, and uncovered a pattern of asynchronous feeding. Altogether, these findings provide valuable descriptive data on the diets of an understudied group of epipelagic teleosts, and, by extension, suggest that prey partitioning (taxa and feeding times) may influence flyingfish feeding ecology by reducing interspecific competition.
Candidaemia in a Polish tertiary paediatric hospital, 2000 to 2010
Authors: Dzierzanowska‐Fangrat Katarzyna , Romanowska Ewa , Gryniewicz‐Kwiatkowska Olga , Migdał Marek , Witulska Katarzyna , Ryżko Józef , Kaliciński Piotr , Książyk Janusz , Nadkowska Paulina , Dzierzanowska Danuta
Published: July 2013

Data on the epidemiology of invasive Candida infections in paediatric patients in Europe are still limited. The aim of this retrospective study was to analyse the epidemiology of candidaemia in a tertiary paediatric hospital in Poland from 2000 to 2010. Using microbiological records, a total of 118 episodes of candidaemia were identified in 114 children, with an annual incidence of 0.35 episodes/1000 discharges. The highest incidences were found in the medical intensive care unit (5.28), and in neonatal intensive care (1.47). The mortality rate was 8.5%. Candida albicans and C. parapsilosis were the most prevalent species (39.8% and 35.6% respectively). The prevalence of non‐albicans species increased from 12.5% in 2000 to 70% in 2010. No differences were found between C. albicans and C. non‐albicans episodes in terms of demographics, risk factors or mortality. The highest resistance rates (overall 7.6%) were observed for fluconazole (4.3% in C. albicans, 7.1% in C. parapsilosis and 13.8% in other Candida species). Resistance to amphotericin B (2.5%) was limited to non‐albicans isolates. The dynamic changes in species distribution and increasing resistance of fungal pathogens confirm the importance of epidemiological surveillance.
Mitochondrial polymorphisms and dysfunction related to aggressive periodontitis: a pilot study
Authors: Wang X , Luan Q , Chen Q , Zhao L , Guo Y
Published: July 2013

Aim

To investigate whether aggressive periodontitis (AgP) is associated with specific mtDNA polymorphisms or point mutations and furthermore whether mitochondrial dysfunction occurs in gingival fibroblasts of AgP patients.

Materials and methods

The mitochondrial DNA coding regions were amplified and sequenced in 22 overlapped fragments in 34 patients with AgP and 28 healthy controls for initial screening. We selected eleven SNPs for detailed investigation in the rest 30 AgP patients and 26 healthy controls, because all other variants occurred at relatively low frequencies or had no difference between two groups. Logistic regression models were used to analyze the association between mtDNA variants and AgP. Gingival fibroblasts were cultured from four patients with AgP and four healthy controls, and then mitochondrial membrane potential, reactive oxygen species production and cell proliferation were analyzed.

Results

Significant association was observed between aggressive periodontitis and eight mitochondrial polymorphisms: ‘8701A‐9540T‐10400C‐10873T‐14783T‐15043G‐15301G’ (OR = 3.471 (1.610–7.483), P = 0.001) and 10398A (OR = 3.238 (1.481–7.078), P = 0.003). Compared with the controls, patients with aggressive periodontitis had a decrease in mitochondrial membrane potential and an increase in reactive oxygen species production.

Conclusion

In conclusion, we propose that mitochondrial dysfunction and ‘8701A‐9540T‐10400C‐10873T‐14783T‐15043G‐15301G, 10398A’ are associated with and may increase the susceptibility to AgP in the Han Chinese population.
Mongolian Spots—A Prospective Study
Authors: Gupta Divya , Thappa Devinder M.
Published: July 2013

To determine the frequency and clinical presentation of Mongolian spots (MS) and assess their evolution with age, this study was conducted in three phases. The first phase examined 2,313 babies born at Jawaharlal Institute of Postgraduate Medical Education and Research between October and December 2010 for the number, size, shape, color, and distribution of MS. Babies with MS were followed up at 6 months and 1 year, in the second and third phases, respectively, to note the evolution of the patches. Of 2,313 babies, 1,524 (65.9%) had MS. The majority had a single patch (n = 790), measuring less than 5 cm (n = 932), with an irregular shape (n = 981) and a blue‐green color (n = 577). The most common site was sacral (n = 1,203), and the most common extrasacral site was a lower extremity (n = 156). A single case of superimposed MS was recorded. Male sex and prematurity were significantly associated with MS (p < 0.05). At 6 months, 73 of 634 babies (11.5%) showed fading and 83 (13.1%) showed complete disappearance. At 1 year, 90 (14.2%) showed fading and 268 (42.3%) showed complete disappearance. Multiple MS (p < 0.05), extrasacral position (p < 0.05), size larger than 10 cm (p < 0.05), and dark‐colored lesions (blue/blue‐black) (p < 0.05) were significantly associated with persistence beyond 1 year. Seven hundred ninety babies (51.8%) had a single MS. More than 40% of MS disappeared by 1 year. Multiple patches, extrasacral position, size larger than 10 cm, and dark‐colored lesions were markers of persistence beyond 1 year.
Failure of bortezomib to cure acute antibody‐mediated rejection in a non‐compliant renal transplant patient
Authors: Ryckewaert A. , Allain‐Launay E. , Moreau A. , Blancho G. , Cesbron A. , Blin N. , Roussey G.
Published: July 2013

Bortezomib has appeared recently as a potential active treatment for acute AMR for few years. We reported a patient who received two courses of bortezomib for the treatment of an acute AMR associated with de novo HLA DSA that occurred 18 months after renal transplantation because of non‐compliance. Graft biopsy revealed features of acute humoral rejection with plasmocyte infiltration and C4d staining. Bortezomib was associated with corticosteroid pulses, IVIgs, and PP. Despite this rapid management, the patient lost his graft and carried on dialysis. Bortezomib therapy in addition to current therapy of AMR is not always effective in the treatment for late acute AMR in renal transplantation. We discuss on the place of such a treatment and other therapeutic strategies in this indication.
Top‐down control of audiovisual search by bimodal search templates
Authors: Matusz Pawel J. , Eimer Martin
Published: July 2013

To test whether the attentional selection of targets defined by a combination of visual and auditory features is guided in a modality‐specific fashion or by control processes that are integrated across modalities, we measured attentional capture by visual stimuli during unimodal visual and audiovisual search. Search arrays were preceded by spatially uninformative visual singleton cues that matched the current target‐defining visual feature. Participants searched for targets defined by a visual feature, or by a combination of visual and auditory features (e.g., red targets accompanied by high‐pitch tones). Spatial cueing effects indicative of attentional capture were reduced during audiovisual search, and cue‐triggered N2pc components were attenuated and delayed. This reduction of cue‐induced attentional capture effects during audiovisual search provides new evidence for the multimodal control of selective attention.
Efficient Estimation of Non-Linear Finite Population Parameters by Using Non-Parametrics
Authors: Goga Camelia , Ruiz-Gazen Anne
Published: July 2013

Currently, high precision estimation of non-linear parameters such as Gini indices, low income proportions or other measures of inequality is particularly crucial. We propose a general class of estimators for such parameters that take into account univariate auxiliary information assumed to be known for every unit in the population. Through a non-parametric model-assisted approach, we construct a unique system of survey weights that can be used to estimate any non-linear parameter that is associated with any study variable of the survey, using a plug-in principle. Based on a rigorous functional approach and a linearization principle, the asymptotic variance of the estimators proposed is derived, and variance estimators are shown to be consistent under mild assumptions. The theory is fully detailed for penalized B-spline estimators together with suggestions for practical implementation and guidelines for choosing the smoothing parameters. The validity of the method is demonstrated on data extracted from the French Labour Force Survey. Point and confidence interval estimation for the Gini index and the low income proportion are derived. Theoretical and empirical results highlight our interest in using a non-parametric versus a parametric approach when estimating non-linear parameters in the presence of auxiliary information.
The Role of Perceived Friendship Self‐efficacy as a Protective Factor against the Negative Effects of Social Victimization
Authors: Fitzpatrick Sally , Bussey Kay
Published: July 2013

This study examined perceived friendship self‐efficacy as a protective factor against the negative effects associated with social victimization in adolescents. The sample consisted of 1218 participants (557 males, age range 12–17 years). Perceived friendship self‐efficacy was associated with lower internalizing scores irrespective of adolescents' social victimization level and with lower externalizing scores at low, but not high, levels of social victimization. Furthermore, the relationship between perceived friendship self‐efficacy and all forms of adjustment did not differ between boys and girls, or between adolescents in both reciprocated and unilateral very best friendships. The role of perceived friendship self‐efficacy as a protective factor amenable to intervention in social bullying at school is discussed.
Improved detection of electrical activity with a voltage probe based on a voltage‐sensing phosphatase
Authors: Tsutsui Hidekazu , Jinno Yuka , Tomita Akiko , Niino Yusuke , Yamada Yoshiyuki , Mikoshiba Katsuhiko , Miyawaki Atsushi , Okamura Yasushi
Published: July 2013

Key points

The use of genetically encoded voltage probes has been expected to enable sensitive detection of spatiotemporal electrical activities in excitable cells.

However, existing probes suffer from low signal amplitude and/or kinetics too slow to detect fast electrical activity.

We have developed an improved voltage probe named Mermaid2.

Mermaid2 provides ratiometric readouts of electrical activity with fast kinetics and great sensitivity, and was able to detect single‐event electrical activity both in vitro and in vivo.

Mermaid2 will expand our chances to analyse electrical events that have been less accessible by using other techniques.

Abstract  One of the most awaited techniques in modern physiology is the sensitive detection of spatiotemporal electrical activity in a complex network of excitable cells. The use of genetically encoded voltage probes has been expected to enable such analysis. However, in spite of recent progress, existing probes still suffer from low signal amplitude and/or kinetics too slow to detect fast electrical activity. Here, we have developed an improved voltage probe named Mermaid2, which is based on the voltage‐sensor domain of the voltage‐sensing phosphatase from Ciona intestinalis and Förster energy transfer between a pair of fluorescent proteins. In mammalian cells, Mermaid2 permits ratiometric readouts of fractional changes of more than 50% over a physiologically relevant voltage range with fast kinetics, and it was used to follow a train of action potentials at frequencies of up to 150 Hz. Mermaid2 was also able to detect single action potentials and subthreshold voltage responses in hippocampal neurons in vitro, in addition to cortical electrical activity evoked by sound stimuli in single trials in living mice.
On the Wave Propagation of Disturbances in Homogeneous Electromechanical Systems
Authors: Dasgupta Kalyan , Kulkarni Anil M. , Soman S. A.
Published: July 2013

In this paper, we make an attempt to describe the phenomenon of wave propagation when a disturbance is introduced in an electromechanical system having a lumped parameter representation. We initially discuss mechanical waves in homogeneous spring mass systems and then focus on electromechanical wave propagation in power systems. We primarily discuss ring and open end systems. Eigenvalue analysis of the system is done to find the behavior of the orthogonal modes as a function of time and space. We then derive an expression for velocity of propagation of the disturbance wave and the transport delay associated with it. Effects of system parameters, like generator inertia and transmission line resistance, are also discussed. Although the theory was developed considering homogeneous systems (identical values of inertia/mass, line parameters/spring constant, etc.), an implementation on a nonhomogeneous system is also presented in this paper. Numerical simulations were done and compared with the analytical results derived in this paper.
Regulators of Calcium Homeostasis Identified by Inference of Kinetic Model Parameters from Live Single Cells Perturbed by siRNA
Authors: Bandara Samuel , Malmersjö Seth , Meyer Tobias
Published: July 2013

Estimation of parameters in a calcium regulatory network from single-cell data illustrates how this approach can reveal protein function.
Immunotherapy Packs a One-Two Punch
Authors: Mueller Kristen L.
Published: July 2013

CD47 antagonists augment the effectiveness of antibody-mediated tumor immunotherapy in mice.
Potassium Controls the Inflammasome
Authors: Foley John F.
Published: July 2013

The common link among the diverse stimuli that activate the NLRP3 inflammasome is the efflux of intracellular potassium ions.
The Polymorphic Linker Domain of
pfmdr1
Is Associated with Resistance-Conferring Mutations in Plasmodium falciparum Populations from East and West Africa
Authors: Okombo John , Zongo Issaka , Gadalla Nahla , Bousema Teun , Beshir Khalid B. , Roper Cally , Hallett Rachel , Ochola-Oyier Lynette Isabella , Sutherland Colin J.
Published: July 2013

Sequence variation in the asparagine/aspartate-rich domain of
pfmdr1
in 215 isolates of
Plasmodium falciparum
from three African countries was compared with published data. The role of this domain in modulating antimalarial sensitivity has not been established. The
pfmdr1
86Y allele was significantly associated with different configurations of the Asn/Asp-rich domain in West and East Africa. In Kenya, a specific form of the Asn/Asp-rich domain was significantly linked to the 86Y, 184Y, and 1246Y haplotype of
pfmdr1
.
Induction of P-Glycoprotein by Antiretroviral Drugs in Human Brain Microvessel Endothelial Cells
Authors: Chan Gary N. Y. , Patel Rucha , Cummins Carolyn L. , Bendayan Reina
Published: July 2013

The membrane-associated drug transporter P-glycoprotein (P-gp) plays an essential role in drug efflux from the brain. Induction of this protein at the blood-brain barrier (BBB) could further affect the ability of a drug to enter the brain. At present, P-gp induction mediated by antiretroviral drugs at the BBB has not been fully investigated. Since P-gp expression is regulated by ligand-activated nuclear receptors, i.e., human pregnane X receptor (hPXR) and human constitutive androstane receptor (hCAR), these receptors could represent potential pathways involved in P-gp induction by antiretroviral drugs. The aims of this study were (i) to determine whether antiretroviral drugs currently used in HIV pharmacotherapy are ligands for hPXR or hCAR and (ii) to examine P-gp function and expression in human brain microvessel endothelial cells treated with antiretroviral drugs identified as ligands of hPXR and/or hCAR. Luciferase reporter gene assays were performed to examine the activation of hPXR and hCAR by antiretroviral drugs. The hCMEC/D3 cell line, which is known to display several morphological and biochemical properties of the BBB in humans, was used to examine P-gp induction following 72 h of exposure to these agents. Amprenavir, atazanavir, darunavir, efavirenz, ritonavir, and lopinavir were found to activate hPXR, whereas abacavir, efavirenz, and nevirapine were found to activate hCAR. P-gp expression and function were significantly induced in hCMEC/D3 cells treated with these drugs at clinical concentrations in plasma. Together, our data suggest that P-gp induction could occur at the BBB during chronic treatment with antiretroviral drugs identified as ligands of hPXR and/or hCAR.
IMP-43 and IMP-44 Metallo-β-Lactamases with Increased Carbapenemase Activities in Multidrug-Resistant Pseudomonas aeruginosa
Authors: Tada Tatsuya , Miyoshi-Akiyama Tohru , Shimada Kayo , Shimojima Masahiro , Kirikae Teruo
Published: July 2013

Two novel IMP-type metallo-β-lactamase variants, IMP-43 and IMP-44, were identified in multidrug-resistant
Pseudomonas aeruginosa
isolates obtained in medical settings in Japan. Analysis of their predicted amino acid sequences revealed that IMP-43 had an amino acid substitution (Val67Phe) compared with IMP-7 and that IMP-44 had two substitutions (Val67Phe and Phe87Ser) compared with IMP-11. The amino acid residue at position 67 is located at the end of a loop close to the active site, consisting of residues 60 to 66 in IMP-1, and the amino acid residue at position 87 forms a hydrophobic patch close to the active site with other amino acids. An
Escherichia coli
strain expressing
bla
IMP-43
was more resistant to doripenem and meropenem but not to imipenem than one expressing
bla
IMP-7
. An
E. coli
strain expressing
bla
IMP-44
was more resistant to doripenem, imipenem and meropenem than one expressing
bla
IMP-11
. IMP-43 had more efficient catalytic activities against all three carbapenems than IMP-7, indicating that the Val67Phe substitution contributed to increased catalytic activities against carbapenems. IMP-44 had more efficient catalytic activities against all carbapenems tested than IMP-11, as well as increased activities compared with IMP-43, indicating that both the Val67Phe and Phe87Ser substitutions contributed to increased catalytic activities against carbapenems.
Disposition of Mefloquine and Enpiroline Is Highly Influenced by a Chronic Schistosoma mansoni Infection
Authors: Ingram Katrin , Duthaler Urs , Vargas Mireille , Ellis William , Keiser Jennifer
Published: July 2013

Chronic
Schistosoma mansoni
infections lead to severe tissue destruction of the gut wall and liver and can influence drug disposition. This study aimed to investigate the impact of a chronic
S. mansoni
infection on the pharmacokinetic (PK) parameters of two promising antischistosomal lead candidates (mefloquine and enpiroline) in mice. Studies were conducted in two different mouse cohorts (
S. mansoni
-infected and uninfected mice) for both drugs. Plasma samples were collected at various time points after oral treatment (200 mg/kg of body weight) with study drugs. A high-performance liquid chromatography (HPLC) method was validated to analyze enpiroline and mefloquine in plasma. Livers and intestines were collected from infected animals to determine the onset of action, hepatic shift, and worm burden reduction. Following mefloquine administration, hepatic shifting and significant worm burden reductions (79.2%) were observed after 72 h. At 1 week posttreatment with enpiroline, the majority of worms had migrated to the liver and significant worm burden reductions were observed (93.1%). The HPLC method was selective, accurate (87.8 to 111.4%), and precise (<10%) for the analysis of both drugs in plasma samples. The PK profiles revealed increased values for half-life (
t
1/2
) and area under the concentration-time curve (AUC) for both drugs in infected animals compared to the
t
1/2
and AUC values in uninfected animals. Considerable changes were observed for mefloquine, with a 5-fold increase of
t
1/2
(182.7 h versus 33.6 h) and 2-fold increase of AUC (1,116,517.8 ng · h/ml versus 522,409.1 ng · h/ml).
S. mansoni
infections in mice influence the PK profiles of enpiroline and mefloquine, leading to delayed clearance. Our data confirm that drug disposition should be carefully studied in schistosomiasis patients.
Screening for Glycosylphosphatidylinositol-Modified Cell Wall Proteins in Pichia pastoris and Their Recombinant Expression on the Cell Surface
Authors: Zhang Li , Liang Shuli , Zhou Xinying , Jin Zi , Jiang Fengchun , Han Shuangyan , Zheng Suiping , Lin Ying
Published: July 2013

Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have various intrinsic functions in yeasts and different uses
in vitro
. In the present study, the genome of
Pichia pastoris
GS115 was screened for potential GPI-modified cell wall proteins. Fifty putative GPI-anchored proteins were selected on the basis of (i) the presence of a C-terminal GPI attachment signal sequence, (ii) the presence of an N-terminal signal sequence for secretion, and (iii) the absence of transmembrane domains in mature protein. The predicted GPI-anchored proteins were fused to an alpha-factor secretion signal as a substitute for their own N-terminal signal peptides and tagged with the chimeric reporters FLAG tag and mature
Candida antarctica
lipase B (CALB). The expression of fusion proteins on the cell surface of
P. pastoris
GS115 was determined by whole-cell flow cytometry and immunoblotting analysis of the cell wall extracts obtained by β-1,3-glucanase digestion. CALB displayed on the cell surface of
P. pastoris
GS115 with the predicted GPI-anchored proteins was examined on the basis of potential hydrolysis of
p
-nitrophenyl butyrate. Finally, 13 proteins were confirmed to be GPI-modified cell wall proteins in
P. pastoris
GS115, which can be used to display heterologous proteins on the yeast cell surface.
Duodenal-Mucosal Bacteria Associated with Celiac Disease in Children
Authors: Sánchez Ester , Donat Ester , Ribes-Koninckx Carmen , Fernández-Murga Maria Leonor , Sanz Yolanda
Published: July 2013

Celiac disease (CD) is an immune-mediated enteropathy triggered by the ingestion of cereal gluten proteins. This disorder is associated with imbalances in the gut microbiota composition that could be involved in the pathogenesis of CD. The aim of this study was to characterize the composition and diversity of the cultivable duodenal mucosa-associated bacteria of CD patients and control children. Duodenal biopsy specimens from patients with active disease on a gluten-containing diet (n= 32), patients with nonactive disease after adherence to a gluten-free diet (n= 17), and controls (n= 8) were homogenized and plated on plate count agar, Wilkins-Chalgren agar, brain heart agar, or yeast, Casitone, and fatty acid agar. The isolates were identified by partial 16S rRNA gene sequencing. Renyi diversity profiles showed the highest diversity values for active CD patients, followed by nonactive CD patients and control individuals. Members of the phylumProteobacteriawere more abundant in patients with active CD than in the other child groups, while those of the phylumFirmicuteswere less abundant. Members of the familiesEnterobacteriaceaeandStaphylococcaceae, particularly the speciesKlebsiella oxytoca,Staphylococcus epidermidis, andStaphylococcus pasteuri, were more abundant in patients with active disease than in controls. In contrast, members of the familyStreptococcaceaewere less abundant in patients with active CD than in controls. Furthermore, isolates of theStreptococcus anginosusandStreptococcus mutansgroups were more abundant in controls than in both CD patient groups, regardless of inflammatory status. The findings indicated that the disease is associated with the overgrowth of possible pathobionts that exclude symbionts or commensals that are characteristic of the healthy small intestinal microbiota.
Occurrence of Waterborne Pathogens and Escherichia coli at Offshore Drinking Water Intakes in Lake Ontario
Authors: Edge T. A. , Khan I. U. H. , Bouchard R. , Guo J. , Hill S. , Locas A. , Moore L. , Neumann N. , Nowak E. , Payment P. , Yang R. , Yerubandi R. , Watson S.
Published: July 2013

The occurrence of waterborne pathogens was investigated at three drinking water intakes located about 2 km offshore in Lake Ontario. Water sampling was conducted over 3 years for
Campylobacter
spp.,
Cryptosporidium
spp.,
Giardia
spp., cultivable enteric viruses, and water quality parameters. All pathogens were detected in the offshore source water for each water treatment plant (WTP1 to WTP3), although at relatively low frequencies and concentrations.
Giardia
was the most common pathogen, occurring in 36% of water samples from the influent of WTP1 (
n
= 46), and with a maximum concentration of 0.70 cysts/liter in this influent.
Cryptosporidium
occurred as frequently as 15% in the WTP2 influent (
n
= 35), with a maximum concentration of 0.40 oocysts/liter in the WTP1 influent. The human
Bacteroidales
HF183 DNA marker was most common in the WTP1 influent (19%), and this was the only WTP where the
Cryptosporidium hominis
genotype was detected. No water quality parameter was predictive of pathogen occurrence across all three WTP influents.
Escherichia coli
was often below detection when pathogens were detected, and spikes in
E. coli
concentrations often did not coincide with pathogen occurrence. After summer rain events, river plumes had
E. coli
concentrations as high as 222 CFU/100 ml in surface waters 2 km offshore, without impacting drinking water intakes below the thermocline on the lake bottom. At times, prechlorination to control mussels at offshore intake cribs compromised the use of
E. coli
for “raw” water quality assessment, particularly for chlorine-resistant
Cryptosporidium. E. coli
measured by standard methods did not reliably predict pathogen occurrence at drinking water intakes in offshore ecosystems.
Diversity, Community Composition, and Dynamics of Nonpigmented and Late-Pigmenting Rapidly Growing Mycobacteria in an Urban Tap Water Production and Distribution System
Authors: Dubrou S. , Konjek J. , Macheras E. , Welté B. , Guidicelli L. , Chignon E. , Joyeux M. , Gaillard J. L. , Heym B. , Tully T. , Sapriel G.
Published: July 2013

Nonpigmented and late-pigmenting rapidly growing mycobacteria (RGM) have been reported to commonly colonize water production and distribution systems. However, there is little information about the nature and distribution of RGM species within the different parts of such complex networks or about their clustering into specific RGM species communities. We conducted a large-scale survey between 2007 and 2009 in the Parisian urban tap water production and distribution system. We analyzed 1,418 water samples from 36 sites, covering all production units, water storage tanks, and distribution units; RGM isolates were identified by usingrpoBgene sequencing. We detected 18 RGM species and putative new species, with most isolates beingMycobacterium chelonaeandMycobacterium llatzerense. Using hierarchical clustering and principal-component analysis, we found that RGM were organized into various communities correlating with water origin (groundwater or surface water) and location within the distribution network. Water treatment plants were more specifically associated with species of theMycobacterium septicumgroup. On average,M. chelonaedominated network sites fed by surface water, andM. llatzerensedominated those fed by groundwater. Overall, theM. chelonaeprevalence index increased along the distribution network and was associated with a correlative decrease in the prevalence index ofM. llatzerense, suggesting competitive or niche exclusion between these two dominant species. Our data describe the great diversity and complexity of RGM species living in the interconnected environments that constitute the water production and distribution system of a large city and highlight the prevalence index of the potentially pathogenic speciesM. chelonaein the distribution network.
Deletion of
creB
in Aspergillus oryzae Increases Secreted Hydrolytic Enzyme Activity
Authors: Hunter A. J. , Morris T. A. , Jin B. , Saint C. P. , Kelly J. M.
Published: July 2013

Aspergillus oryzae
has been used in the food and beverage industry for centuries, and industrial strains have been produced by multiple rounds of selection. Targeted gene deletion technology is particularly useful for strain improvement in such strains, particularly when they do not have a well-characterized meiotic cycle. Phenotypes of an
Aspergillus nidulans
strain null for the CreB deubiquitinating enzyme include effects on growth and repression, including increased activity levels of various enzymes. We show that
Aspergillus oryzae
contains a functional homologue of the CreB deubiquitinating enzyme and that a null strain shows increased activity levels of industrially important secreted enzymes, including cellulases, xylanases, amylases, and proteases, as well as alleviated inhibition of spore germination on glucose medium. Reverse transcription-quantitative PCR (RT-qPCR) analysis showed that the increased levels of enzyme activity in both
Aspergillus nidulans
and
Aspergillus oryzae
are mirrored at the transcript level, indicating transcriptional regulation. We report that
Aspergillus oryzae
DAR3699, originally isolated from soy fermentation, has a similar phenotype to that of a
creB
deletion mutant of the RIB40 strain, and it contains a mutation in the
creB
gene. Collectively, the results for
Aspergillus oryzae
,
Aspergillus nidulans
,
Trichoderma reesei
, and
Penicillium decumbens
show that deletion of
creB
may be broadly useful in diverse fungi for increasing production of a variety of enzymes.
Structure-Function Relationships in Hydrophobins: Probing the Role of Charged Side Chains
Authors: Lienemann Michael , Gandier Julie-Anne , Joensuu Jussi J. , Iwanaga Atsushi , Takatsuji Yoshiyuki , Haruyama Tetsuya , Master Emma , Tenkanen Maija , Linder Markus B.
Published: July 2013

Hydrophobins are small fungal proteins that are amphiphilic and have a strong tendency to assemble at interfaces. By taking advantage of this property, hydrophobins have been used for a number of applications: as affinity tags in protein purification, for protein immobilization, such as in foam stabilizers, and as dispersion agents for insoluble drug molecules. Here, we used site-directed mutagenesis to gain an understanding of the molecular basis of their properties. We especially focused on the role of charged amino acids in the structure of hydrophobins. For this purpose, fusion proteins consisting of
Trichoderma reesei
hydrophobin I (HFBI) and the green fluorescent protein (GFP) that contained various combinations of substitutions of charged amino acids (D30, K32, D40, D43, R45, K50) in the HFBI structure were produced. The effects of the introduced mutations on binding, oligomerization, and partitioning were characterized in an aqueous two-phase system. It was found that some substitutions caused better surface binding and reduced oligomerization, while some showed the opposite effects. However, all mutations decreased partitioning in surfactant systems, indicating that the different functions are not directly correlated and that partitioning is dependent on finely tuned properties of hydrophobins. This work shows that not all functions in self-assembly are connected in a predictable way and that a simple surfactant model for hydrophobin function is insufficient.
Augmenting the Genetic Toolbox for Sulfolobus islandicus with a Stringent Positive Selectable Marker for Agmatine Prototrophy
Authors: Zhang Changyi , Cooper Tara E. , Krause David J. , Whitaker Rachel J.
Published: July 2013

Sulfolobus
species have become the model organisms for studying the unique biology of the crenarchaeal division of the archaeal domain. In particular,
Sulfolobus islandicus
provides a powerful opportunity to explore natural variation via experimental functional genomics. To support these efforts, we further expanded genetic tools for
S. islandicus
by developing a stringent positive selection for agmatine prototrophs in strains in which the
argD
gene, encoding arginine decarboxylase, has been deleted. Strains with deletions in
argD
were shown to be auxotrophic for agmatine even in nutrient-rich medium, but growth could be restored by either supplementation of exogenous agmatine or reintroduction of a functional copy of the
argD
gene from
S. solfataricus
P2 into the Δ
argD
host. Using this stringent selection, a robust targeted gene knockout system was established via an improved next generation of the MID (
m
arker
i
nsertion and unmarked target gene
d
eletion) method. Application of this novel system was validated by targeted knockout of the
upsEF
genes involved in UV-inducible cell aggregation formation.