bluuebunny/crossref_metadata_2025 huggingface dataset:
Validation of the Dutch Version of the DN4 Diagnostic Questionnaire for Neuropathic Pain
Authors: van Seventer Robert , Vos Cornelis , Giezeman Maurice , Meerding Willem‐Jan , Arnould Benoit , Regnault Antoine , van Eerd Maarten , Martin Carola , Huygen Frank
Published: November 2012

Difficulties in diagnosing neuropathic pain in routine clinical practice support the need for validated and easy‐to‐use diagnostic tools. The DN4 neuropathic pain diagnostic questionnaire aims to discriminate neuropathic pain from nociceptive pain, but needs clinical validation. A total of 269 patients with chronic pain in three pain clinics were included in the study of which 248 had analyzable data. The mean duration of pain was 4.9 years. The most frequent etiologies were posttraumatic (36%), (pseudo) radicular (14%), and mechanical back pain (12%). The mean intensity of pain at visit was 5.6 on a 0–10 scale. Hundred and ninety‐six of 248 patients had an identical pain diagnosis from both physicians: 85 had neuropathic pain, 57 had nociceptive pain, and 54 had mixed pain. Among patients with identical diagnoses of neuropathic or nociceptive pain, using a receiver operating characteristic curve analysis, the area under the curve (AUC) was 0.81 for the DN4 7‐item and 0.82 for the 10‐item version. A cutoff point of 5/10 for the full questionnaire resulted in a sensitivity of 75% and a specificity of 79%, while a cutoff point of 4/7 for the partial questionnaire resulted in a sensitivity of 74% and a specificity of 79%. The items “brushing,” “painful cold,” and “numbness” were most discriminating. The DN4 is an easy‐to‐use screening tool that is reliable for discriminating between neuropathic and nociceptive pain conditions in daily practice. Item‐specific scores provide important information in addition to the total score.
The Influence of the Antiandrogen 2‐Hydroxyflutamide on the Androgen Receptor Expression in the Porcine Ovarian Follicles – An In Vitro Study
Authors: Duda M , Wolna A , Knapczyk‐Stwora K , Durlej‐Grzesiak M , Knet M , Tabarowski Z , Slomczynska M
Published: November 2012

Contents

Androgens are one of the most important agents influencing ovarian follicles growth and development. The biological action of androgens is primarily exerted through transcriptional regulation by the androgen receptor (AR), a member of the steroid hormone receptor superfamily. The purpose of this study was to test the role of androgen receptor agonist testosterone (T) or antagonist 2‐hydroxyflutamide (2‐Hf) and in combination on AR expression in cultured porcine granulosa cells (GC) or whole follicles. Granulosa cells isolated from mature pig follicles were cultured for 48 h. During the last 12 and 24 h of culture, they were incubated in the presence of T (10−7 m/ml), 2‐Hf (1.7 × 10−4 m) or both T and 2‐Hf (T + 2‐Hf, at the same concentrations as when added separately). To better imitate in vivo conditions, whole follicles (6–8 mm in diameter) isolated from porcine ovaries have been incubated (for 12 and 24 h) in an organ culture system with the addition of the same factors. Thereafter, cells or sections obtained from cultured follicles were processed for AR detection by immunocytochemistry or immunohistochemistry. Moreover, expression of AR mRNA and protein was determined by real‐time PCR and Western blot analysis. It was shown that the addition of 2‐Hf in the presence of T had a positive effect on AR mRNA and protein expression in porcine GC and ovarian follicles. Moreover, the addition of 2‐Hf influenced AR distribution in GC cultures which is seen as change of its localization from nuclear to perinuclear. Our results suggest that androgens acting through AR could be involved in the control of AR expression in porcine GC in vitro and in vivo.
Decreased activities of daily living is a strong risk factor for liver injury by anti‐tuberculosis drugs
Authors: Horita Nobuyuki , Miyazawa Naoki , Yoshiyama Takashi , Tsukahara Toshinori , Takahashi Ryohei , Tsukiji Jun , Kato Hideaki , Kaneko Takeshi , Ishigatsubo Yoshiaki
Published: November 2012

Background and objective

We evaluated the association between activities of daily living and drug‐induced liver injury by anti‐tuberculosis drugs.

Methods

This retrospective cohort study included adult inpatients with newly diagnosed smear‐positive lung tuberculosis treated with standard regimen in two hospitals. (n = 346; 63.6 ± 20.3 years old; 106 (30.6%) females). Activities of daily living was divided into ‘independent’ (Barthel Index (BI) 80–100, 60.4%) and ‘decreased’ (BI 0–75, 39.6%) categories. Liver injury was defined as the withdrawal or change of treatment regimen on the basis of the following criteria: serum transaminase concentrations were more than three times the upper limit of normal range with jaundice and/or hepatitis symptoms, or more than five times the upper limit of the normal range.

Results

Compared with ‘independent’ patients, patients with ‘decreased’ activities of daily living had odds ratios for liver injury of 4.2 (P < 0.001) in univariate analysis and 5.7 (P = 0.002) in logistic regression analysis after adjusting for other risk factors.

Conclusions

Decreased activity of daily living is a strong risk factor for liver injury among adult inpatients with newly diagnosed smear‐positive lung tuberculosis treated using a standard regimen.
Ambulatory oximetry fails to predict survival in chronic obstructive pulmonary disease with mild‐to‐moderate hypoxaemia
Authors: TRAUER JAMES M. , GIELEN CHELSEA A. , AMINAZAD ALI , Steinfort Christopher L.
Published: November 2012

Background and objective

Although long‐term oxygen therapy is one of few treatments known to improve survival in COPD, no patient subgroup likely to derive benefit has been identified in over 30 years. We assessed the prognostic value of point measurement of PaO2 and proportion of ambulatory oximetry with saturations below 90% in this context.

Methods

Subjects were community‐living patients with COPD and PaO2 56–70 mm Hg when stable. Baseline assessments included clinical, physiological and laboratory assessments, as well as 24‐h ambulatory oximetry. Patients were followed to death from any cause, first exacerbation and first admission.

Results

Despite poor survival overall, there was no significant difference in prognosis between mildly hypoxaemic and moderately hypoxaemic patients. There were also no significant differences for secondary end–points (exacerbation and hospital admission). On multivariate analysis, trends were observed towards improved survival, with higher PaO2 and greater proportion of ambulatory oximetry below 90%.

Conclusions

Neither resting PaO2 nor proportion of ambulatory oximetry below 90% saturation effectively predicted survival in COPD.
Sonographic assessment of changes in diaphragmatic kinetics induced by inspiratory resistive loading
Authors: Soilemezi Eleni , Tsagourias Matthew , Talias Michael A. , Soteriades Elpidoforos S. , Makrakis Vasilios , Zakynthinos Epaminondas , Matamis Dimitrios
Published: November 2012

Background and objective

Diaphragmatic breathing patterns under resistive loading remain poorly documented. To our knowledge, this is the first study assessing diaphragmatic motion under conditions of inspiratory resistive loading with the use of sonography.

Methods

We assessed diaphragmatic motion during inspiratory resistive loading in 40 healthy volunteers using M‐mode sonography. In phase I of the study, sonography was performed during normal quiet breathing without respiratory loading. In phase II, sonography was performed after application of a nose clip and connection of the subjects to a pneumotachograph through a mouth piece. In phase III, the participants were assessed while subjected to inspiratory resistive loading of 50 cm H2O/L/s.

Results

Compared with baseline, the application of a mouth piece and nose clip induced a significant increase in diaphragmatic excursion (from 1.7 to 2.3 cm, P < 0.001) and a decrease in respiratory rate (from 13.4 to 12.2, P < 0.01). Inspiratory resistive loading induced a further decrease in respiratory rate (from 12.2 to 8.0, P < 0.01) and a decrease in diaphragmatic velocity contraction (from 1.2 to 0.8 cm/s, P < 0.01), and also an increase in tidal volume (from 795 to 904 mL, P < 0.01); diaphragmatic excursion, however, did not change significantly.

Conclusions

Inspiratory resistive loading induced significant changes in diaphragmatic contraction pattern, which mainly consisted of decreased velocity of diaphragmatic displacement with no change in diaphragmatic excursion. Tidal volume, increased significantly; the increase in tidal volume, along with the unchanged diaphragmatic excursion, provides sonographic evidence of increased recruitment of extradiaphragmatic muscles under inspiratory resistive loading.
Self‐transcendence, nurse–patient interaction and the outcome of multidimensional well‐being in cognitively intact nursing home patients
Authors: Haugan Gørill , Hanssen Brith , Moksnes Unni K.
Published: November 2012

Aims and objectives

The aim of this study was to investigate the associations between age, gender, self‐transcendence, nurse–patient interaction and multidimensional well‐being as the outcome among cognitively intact nursing home patients.

Background

Self‐transcendence is considered to be a vital resource of well‐being in vulnerable populations and at the end of life. Moreover, the quality of care and the nurse–patient interaction is found to influence self‐transcendence and well‐being in nursing home patients.

Design and method

A cross‐sectional design employing the Self‐Transcendence Scale, the Nurse–Patient Interaction Scale, the FACT‐G Quality of Life and the FACIT‐Sp Spiritual Well‐Being questionnaires was adopted. A sample of 202 cognitively intact nursing home patients from 44 nursing homes in central Norway was selected. A previous documented two‐factor construct of self‐transcendence was applied. The statistical analyses were carried out by means of independent sample t‐test, correlation and regression analyses.

Results

Multiple linear regression analyses revealed significant relationships between interpersonal self‐transcendence and social, functional and spiritual well‐being, whereas intrapersonal self‐transcendence significantly related to emotional, social, functional and spiritual well‐being. Nurse–patient interaction related to physical, emotional and functional well‐being. Age and gender were not significant predictors for well‐being, except for functional and spiritual well‐being where women scored higher than men.

Conclusion

Nurse–patient interaction and self‐transcendence are vital resources for promoting well‐being physically, emotionally, functionally, socially and spiritually among cognitively intact nursing home patients.

Relevance to clinical practice

Nurse–patient interaction signifies vital and ultimate nursing qualities promoting self‐transcendence and multidimensional well‐being. These findings are important for clinical nursing intending to increase patients’ well‐being.
Male swimmers cross theEnglishChannel faster than female swimmers
Authors: Fischer G. , Knechtle B. , Rüst C. A. , Rosemann T.
Published: November 2012

We examined the gender difference in performance of open‐water ultra‐swimmers crossing theEnglishChannel between 1875 and 2011. A total of 1606 swimmers (1120 males and 486 females) crossed theEnglishChannel within a mean time of 809.6 ± 175.6 min. The overall female swim time of 796.3 ± 188.7 min was not different compared with the overall male swim time of 815.4 ± 169.4 min (P > 0.05). The fastest male swim time ever of 417 min was 6.7% faster than the fastest female swim time ever with 445 min. The gender difference in performance of the top three times ever was 8.9 ± 2.3%. Over the last 36 years, the performance of the annual top three swimmers showed no changes for both females and males. The top three males (564.3 ± 63.8 min) were significantly faster than the top three females (602.1 ± 58.7 min;P < 0.01). The gender difference remained unchanged at 12.5 ± 9.6% over the years. To summarize, the top three male swimmers in theEnglishChannel were ∼12% faster than the females in the last 36 years. It seems unlikely that female open‐water ultra‐swimmers will overtop males in the near future in theEnglishChannel.
Sex Differences in the Relationship between Harsh Discipline and Conduct Problems
Authors: Lysenko Laura J. , Barker Edward D. , Jaffee Sara R.
Published: November 2012

Research on sex differences in antisocial behaviour may shed light on the causes of childhood antisocial behaviour. Using a longitudinal design, we tested whether there were sex differences in the amount of harsh discipline children received or in the effect of harsh discipline and whether this accounted for sex differences in later conduct problems. Our sample was a representative, longitudinal sample of 13 830 twins born in England and Wales between 1994 and 1996. Results showed that boys experienced more harsh discipline than girls and that the sex difference in harsh discipline accounted for 10 percent to 20 percent of the sex difference in conduct problems. We found no evidence that harsh discipline had a greater effect on boys vs. girls. We also found evidence of a bidirectional relationship between harsh discipline and child conduct problems. These findings were replicated within families, automatically controlling for between‐families confounding factors.
Nuclear Import of Chromatin Remodeler Isw1 Is Mediated by Atypical Bipartite cNLS and Classical Import Pathway
Authors: Vasicova Pavla , Stradalova Vendula , Halada Petr , Hasek Jiri , Malcova Ivana
Published: November 2012

The protein Isw1 of Saccharomyces cerevisiae is an imitation‐switch chromatin‐remodeling factor. We studied the mechanisms of its nuclear import and found that the nuclear localization signal (NLS) mediating the transport of Isw1 into the nucleus is located at the end of the C‐terminus of the protein (aa1079–1105). We show that it is an atypical bipartite signal with an unconventional linker of 19 aa (KRIR X19 KKAK) and the only nuclear targeting signal within the Isw1 molecule. The efficiency of Isw1 nuclear import was found to be modulated by changes to the amino acid composition in the vicinity of the KRIR motif, but not by the linker length. Live‐cell imaging of various karyopherin mutants and in vitro binding assays of Isw1NLS to importin‐α revealed that the nuclear translocation of Isw1 is mediated by the classical import pathway. Analogous motifs to Isw1NLS are highly conserved in Isw1 homologues of other yeast species, and putative bipartite cNLS were identified in silico at the end of the C‐termini of imitation switch (ISWI) proteins from higher eukaryotes. We suggest that the C‐termini of the ISWI family proteins play an important role in their nuclear import.
Actinomyces bowdenii ulcerative keratitis in a dog
Authors: Sherman Amanda , Daniels Joshua B. , Wilkie David A. , Lutz Elizabeth
Published: November 2012

A 5‐year‐old spayed female diabetic mixed‐breed dog underwent phacoemulsification and intraocular lens implantation to correct bilateral hypermature cataracts. Two months postsurgery, the patient presented with ulcerative keratitis and multifocal stromal abscessation OD, which was controlled, but never resolved, with topical fluoroquinolone therapy. The patient re‐presented 2 months later with a new, raised, white gritty corneal opacity associated with hyperemia, chemosis, and blepharospasm OD. Cytology of the right cornea revealed filamentous bacteria, suggestive of Actinomyces spp. Actinomyces bowdenii was subsequently isolated in pure culture and identified via 16s rDNA sequencing. Actinomyces bowdenii has never before been described as a cause of ocular infection. An immunosuppressed corneal environment likely contributed to this opportunistic Actinomycosis. The infection was not controlled with fluoroquinolone therapy, and the isolate, in vitro, was resistant to three fluoroquinolones (ciprofloxacin, ofloxacin, and levofloxacin), which also has not been previously reported for this species of Actinomyces. A superficial keratectomy with conjunctival graft was employed to successfully manage the infection.
Conjunctival squamous cell carcinoma in a reindeer (Rangifer tarandus tarandus)
Authors: Gonzalez‐Alonso‐Alegre Elisa M. , Rodriguez‐Alvaro Alfonso , Martinez‐Nevado Eva , Martinez‐de‐Merlo Elena M. , Sanchez‐Maldonado Belen
Published: November 2012

An 8‐year‐old female adult reindeer (Rangifer tarandus tarandus) was referred to the Veterinary Hospital of Madrid for evaluation of a conjunctival mass on the left eye which had been present for about 2 months. A surgical excision was performed and biopsy material submitted for light microscopic evaluation which confirmed the diagnosis of conjuctival squamous cell carcinoma. Nuclear p53 immunolabeling was found in 52% of the neoplastic cells. Follow‐up examination at 12 months postsurgery did not reveal recurrence of this neoplasm. Conjunctival squamous cell carcinoma has not been reported previously in reindeer and seems to have similar characteristics to the one existing in bovine species.
Schwarzian differential equations and Hecke eigenforms on Shimura curves
Authors: Yang Yifan
Published: November 2012

Let X be a Shimura curve of genus zero. In this paper, we first characterize the spaces of automorphic forms on X in terms of Schwarzian differential equations. We then devise a method to compute Hecke operators on these spaces. An interesting by-product of our analysis is the evaluation and other similar identities.
Proof of a positivity conjecture of M. Kontsevich on non-commutative cluster variables
Authors: Lee Kyungyong , Schiffler Ralf
Published: November 2012

We prove a conjecture of Kontsevich, which asserts that the iterations of the non-commutative rational map Fr:(x,y)→(xyx−1,(1+yr)x−1) are given by non-commutative Laurent polynomials with non-negative integer coefficients.
Representations of metaplectic groups I: epsilon dichotomy and local Langlands correspondence
Authors: Gan Wee Teck , Savin Gordan
Published: November 2012

Using theta correspondence, we classify the irreducible representations ofMp2nin terms of the irreducible representations ofSO2n+1and determine many properties of this classification. This is a local Shimura correspondence which extends the well-known results of Waldspurger forn=1.
Complete addition laws on abelian varieties
Authors: Arene Christophe , Kohel David , Ritzenthaler Christophe
Published: November 2012

We prove that under any projective embedding of an abelian variety A of dimension g, a complete set of addition laws has cardinality at least g+1, generalizing a result of Bosma and Lenstra for the Weierstrass model of an elliptic curve in ℙ2. In contrast, we prove, moreover, that if k is any field with infinite absolute Galois group, then there exists for every abelian variety A/k a projective embedding and an addition law defined for every pair of k-rational points. For an abelian variety of dimension 1 or 2, we show that this embedding can be the classical Weierstrass model or the embedding in ℙ15, respectively, up to a finite number of counterexamples for ∣k∣≤5 .
Secondary Flow Loss Reduction Through Blowing for a High-Lift Front-Loaded Low Pressure Turbine Cascade
Authors: Benton Stuart I. , Bons Jeffrey P. , Sondergaard Rolf
Published: November 2012

Efforts to increase individual blade loading in the low pressure turbine have resulted in blade geometries optimized for midspan performance. Many researchers have shown that increased blade loading and a front-loaded pressure distribution each separately contribute to increased losses in the endwall region. A detailed investigation of the baseline endwall flow of the L2F profile, which is a high-lift front loaded profile, is performed. In-plane velocity vectors and total pressure loss maps are obtained in five planes oriented normal to the blade surface for three Reynolds numbers. A row of pitched and skewed jets are introduced near the endwall on the suction surface of the blade. The flow control method is evaluated for four momentum coefficients at the high Reynolds number, with a maximum reduction of 42% in the area averaged total pressure loss coefficient. The same blade is also fitted with midspan vortex-generator jets and is tested at a Reynolds number of 20,000, resulting in a 21% reduction in the area averaged total pressure loss.
Marine Microbes See a Sea of Gradients
Authors: Stocker Roman
Published: November 2012

Ocean Monarchs

It is hard to grasp that the unseen microorganisms of the oceans are the most productive on the planet, at orders of magnitude greater than sharks and whales or even terrestrial forests. The plankton is thus a major contributor to the geochemical cycles that are currently under pressure from climate change.
Stocker
(p.
628
) reviews the state of knowledge of the web of myriad ephemeral microenvironments within the ocean's bulk and how microorganisms respond to the ever-shifting chemical spectrum. To this end,
Taylor and Stocker
(p.
675
) report experiments on the effects of turbulence on nutrient uptake by chemotactic marine bacteria. They propose that turbulence favors motile bacteria that adopt an optimal foraging strategy, which trades off the relative high cost of motility to gain the benefits of plumes of nutrients by zipping between them at optimized speeds. Scaled up, such apparently “micro” behavior will influence the rate of remineralization of dissolved organic matter and in turn will feed into global patterns of geochemical cycling.
Quantum Entanglement of High Angular Momenta
Authors: Fickler Robert , Lapkiewicz Radek , Plick William N. , Krenn Mario , Schaeff Christoph , Ramelow Sven , Zeilinger Anton
Published: November 2012

Twist and Entangle

Entanglement is a key feature in quantum information science and plays an important role in various applications of quantum mechanics.

Fickler
et al.

(p.
640
) present a method for converting the polarization state of photons into information encoded into spatial modes of a single photon. From this, superposition states and entangled photons with very high orbital angular momentum quantum numbers were generated.
Optimizing Investments in Malaria Treatment and Diagnosis
Authors: Cohen Justin M. , Woolsey Aaron M. , Sabot Oliver J. , Gething Peter W. , Tatem Andrew J. , Moonen Bruno
Published: November 2012

Better targeting of antimalarials to people who need them will maximize the impact of interventions in the private sector.
Templating a Molecular Tug-of-War
Authors: Diehl Michael R.
Published: November 2012

Tying cytoskeletal motors together on a DNA rope reveals how opposing motors compete to determine the direction of cargo transport.
Obama and the Promotion of International Science
Authors: Bollyky Thomas J. , Bollyky Paul L.
Published: November 2012

Has the U.S. president's record on promoting international science fulfilled the promise of his early rhetoric?
Excellence,
Ja
, Elitism,
Non
Authors: Macilwain Colin
Published: November 2012

France and Germany are pursuing parallel initiatives to bolster their best universities. Do they go too far or not far enough?
Caspofungin Kills Candida albicans by Causing both Cellular Apoptosis and Necrosis
Authors: Hao Binghua , Cheng Shaoji , Clancy Cornelius J. , Nguyen M. Hong
Published: November 2012

Caspofungin exerts candidacidal activity by inhibiting cell wall (1,3)-β-
d
-glucan synthesis. We investigated the physiologic mechanisms of caspofungin-induced
Candida albicans
cell death. Apoptosis (programmed cell death) and necrosis were studied after
C. albicans
SC5314 cells were exposed to caspofungin at 0.06, 0.125, and 0.5 μg/ml (0.5×, 1×, and 4× the MIC, respectively) for 3 h. Caspofungin at 0.125 and 0.5 μg/ml reduced cellular viability by >50%, as measured by colony counts and methylene blue exclusion. Apoptosis and necrosis were demonstrated by annexin V and propidium iodide staining for phosphatidylserine externalization and loss of membrane integrity, respectively. At all concentrations of caspofungin, 20 to 25% and 5 to 7% of
C. albicans
cells exhibited early apoptosis and late apoptosis/necrosis, respectively (
P
value was not significant [NS]). Necrosis, on the other hand, was significantly greater at 0.125 (43%) and 0.5 (48%) μg/ml than at 0.06 μg/ml (26%) (
P
values of 0.003 and 0.003, respectively). The induction of apoptosis at concentrations less than or equal to the MIC was corroborated by dihydrorhodamine 123 (DHR-123) and dihydroethidium (DHE) staining (reactive oxygen species production), JC-1 staining (mitochondrial membrane potential dissipation), and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) staining (DNA damage and nuclear fragmentation). Moreover, electron microscopy of cells exposed to 0.125 μg/ml of caspofungin showed hallmark apoptotic features like chromatin margination and condensation and nuclear blebs. Apoptosis was associated with metacaspase 1 activation, as demonstrated by D2R staining. Caspofungin exerts activity against
C. albicans
by directly killing cells (resulting in necrosis) and causing others to undergo programmed cell death (apoptosis). Apoptosis is initiated at subinhibitory concentrations, suggesting that strategies to target this process may augment the benefits of antifungal agents.
Benznidazole-Related Adverse Drug Reactions and Their Relationship to Serum Drug Concentrations in Patients with Chronic Chagas Disease
Authors: Pinazo María-Jesús , Guerrero Laura , Posada Elizabeth , Rodríguez Elena , Soy Dolors , Gascon Joaquim
Published: November 2012

For treating Chagas disease (CD), a current worldwide health problem, only benznidazole and nifurtimox have been approved to be used. In both cases, unwanted drug-related adverse events (ADRs) are frequent when these drugs are used in adults in the chronic stage. The main objective of this study was to establish benznidazole ADRs and their relationship to serum concentrations in patients with chronic
Trypanosoma cruzi
infection in order to perform more accurate dosages to minimize ADRs. A total of 54 patients were recruited over 12 months. Of these 54 patients, 53 (98%) experienced at least one ADR during follow-up, and the overall average ADR incidence was 2.4 episodes/patient/month. Benznidazole treatment was discontinued in 11 patients, 7 among them due to severe adverse effects. The mean duration of treatment before withdrawal was 11 days. Benznidazole serum concentrations were recorded on days 15, 30, 45, and 60 of follow-up and evaluated according to clinical and epidemiological variables and ADR severity. No relationship was found between the benznidazole serum concentration and the ADRs. The mean (standard deviation) trough serum benznidazole concentrations (all below 20 mcg/ml) on days 15, 30, 45, and 60 were 6.4 (1.9), 6.1 (1.8), 6.2 (2.2), and 5.7 (1.7) μg/ml, respectively. Benznidazole serum concentrations do not appear to be related to the appearance of serious ADRs. Further, well-controlled studies are necessary to establish the optimal regimen for benznidazole in adults with chronic CD.
Highly Variable Plasma Concentrations of Voriconazole in Pediatric Hematopoietic Stem Cell Transplantation Patients
Authors: Bartelink Imke H. , Wolfs Tom , Jonker Martine , de Waal Marjolein , Egberts Toine C. G. , Ververs Tessa T. , Boelens Jaap J. , Bierings Marc
Published: November 2012

Invasive fungal infections are of great concern in pediatric hematopoietic stem cell transplantation (HSCT) recipients. Voriconazole is usually the drug of first choice for treating or preventing invasive aspergillosis. Optimum trough levels (
C
trough
s) are between 1 and 5 mg/liter. It is unclear whether these levels are reached with currently advised pediatric dosing schedules. Between 2007 and 2011, 11 patients <2 years of age, 31 between 2 and 12 years, and 20 between 12 and 20 years were (prophylactically or therapeutically) treated with voriconazole in the HSCT unit of UMC Utrecht. For children <2 years of age, the dosage recommended for 2 to 12 years was used. In 34% of children who started with the recommended dose, an adequate
C
trough
was reached irrespective of age or administration route. After therapeutic drug monitoring (TDM)-based dose adjustments, adequate
C
trough
s were reached in 80% of the patients at median doses of 31.5 (age, <2 years), 16 (age, 2 to 12 years), and 9.4 mg/kg of body weight/day (age, >12 years) (
P
= 0.034). The intrapatient variability in
C
trough
ranged between 1 and 238%. Voriconazole was discontinued in six patients due to toxicity. These patients had a median
C
trough
of 0.5 mg/liter at the initial dose (ranging from 0.5 to 2.6 mg/liter), and a medium maximal concentration of 4 mg/liter was reached. Inter- and intrapatient variability is a major concern in voriconazole treatment and necessitates therapeutic drug monitoring of dosing, especially in young children.
An Improved Small-Molecule Inhibitor of FtsZ with Superior
In Vitro
Potency, Drug-Like Properties, and
In Vivo
Efficacy
Authors: Stokes Neil R. , Baker Nicola , Bennett James M. , Berry Joanne , Collins Ian , Czaplewski Lloyd G. , Logan Alastair , Macdonald Rebecca , MacLeod Leanne , Peasley Hilary , Mitchell Jeffrey P. , Nayal Narendra , Yadav Anju , Srivastava Anil , Haydon David J.
Published: November 2012

The bacterial cell division protein FtsZ is an attractive target for small-molecule antibacterial drug discovery. Derivatives of 3-methoxybenzamide, including compound PC190723, have been reported to be potent and selective antistaphylococcal agents which exert their effects through the disruption of intracellular FtsZ function. Here, we report the further optimization of 3-methoxybenzamide derivatives towards a drug candidate. The
in vitro
and
in vivo
characterization of a more advanced lead compound, designated compound 1, is described. Compound 1 was potently antibacterial, with an average MIC of 0.12 μg/ml against all staphylococcal species, including methicillin- and multidrug-resistant
Staphylococcus aureus
and
Staphylococcus epidermidis
. Compound 1 inhibited an
S. aureus
strain carrying the G196A mutation in FtsZ, which confers resistance to PC190723. Like PC190723, compound 1 acted on whole bacterial cells by blocking cytokinesis. No interactions between compound 1 and a diverse panel of antibiotics were measured in checkerboard experiments. Compound 1 displayed suitable
in vitro
pharmaceutical properties and a favorable
in vivo
pharmacokinetic profile following intravenous and oral administration, with a calculated bioavailability of 82.0% in mice. Compound 1 demonstrated efficacy in a murine model of systemic
S. aureus
infection and caused a significant decrease in the bacterial load in the thigh infection model. A greater reduction in the number of
S. aureus
cells recovered from infected thighs, equivalent to 3.68 log units, than in those recovered from controls was achieved using a succinate prodrug of compound 1, which was designated compound 2. In summary, optimized derivatives of 3-methoxybenzamide may yield a first-in-class FtsZ inhibitor for the treatment of antibiotic-resistant staphylococcal infections.
Molecular Identification, Antifungal Susceptibility Profile, and Biofilm Formation of Clinical and Environmental Rhodotorula Species Isolates
Authors: Nunes Jorge Meneses , Bizerra Fernando César , Ferreira Renata Carmona e , Colombo Arnaldo Lopes
Published: November 2012

Rhodotorula
species are emergent fungal pathogens capable of causing invasive infections, primarily fungemia. They are particularly problematic in immunosuppressed patients when using a central venous catheter. In this study, we evaluated the species distribution of 51 clinical and 8 environmental
Rhodotorula
species isolates using the ID32C system and internal transcribed spacer (ITS) sequencing. Antifungal susceptibility testing and biofilm formation capability using a crystal violet staining assay were performed. Using ITS sequencing as the gold standard, the clinical isolates were identified as follows: 44
R. mucilaginosa
isolates, 2
R. glutinis
isolates, 2
R. minuta
isolates, 2
R. dairenensis
isolates, and 1
Rhodosporidium fluviale
isolate. The environmental isolates included 7
R. mucilaginosa
isolates and 1
R. slooffiae
isolate. Using the ID32C system, along with a nitrate assimilation test, only 90.3% of the isolates tested were correctly identified. In the biofilm formation assay,
R. mucilaginosa
and
R. minuta
exhibited greater biofilm formation ability compared to the other
Rhodotorula
species; the clinical isolates of
R. mucilaginosa
showed greater biofilm formation compared to the environmental isolates (
P
= 0.04). Amphotericin B showed good
in vitro
activity (MIC ≤ 1 μg/ml) against planktonic cells, whereas voriconazole and posaconazole showed poor activity (MIC
50
/MIC
90
, 2/4 μg/ml). Caspofungin and fluconazole MICs were consistently high for all isolates tested (≥64 μg/ml and ≥ 4 μg/ml, respectively). In this study, we emphasized the importance of molecular methods to correctly identify
Rhodotorula
species isolates and non-
R. mucilaginosa
species in particular. The antifungal susceptibility profile reinforces amphotericin B as the antifungal drug of choice for the treatment of
Rhodotorula
infections. To our knowledge, this is the first study evaluating putative differences in the ability of biofilm formation among different
Rhodotorula
species.
Complete Nucleotide Sequences of
bla
KPC-4
- and
bla
KPC-5
-Harboring IncN and IncX Plasmids from Klebsiella pneumoniae Strains Isolated in New Jersey
Authors: Chen Liang , Chavda Kalyan D. , Fraimow Henry S. , Mediavilla José R. , Melano Roberto G. , Jacobs Michael R. , Bonomo Robert A. , Kreiswirth Barry N.
Published: November 2012

Klebsiella pneumoniae
carbapenemase (KPC)-producing
Enterobacteriaceae
have emerged as major nosocomial pathogens.
bla
KPC
, commonly located on Tn
4401
, is found in Gram-negative bacterial strains, with the two most common variants,
bla
KPC-2
and
bla
KPC-3
, identified in plasmids with diverse genetic backgrounds. In this study, we examined
bla
KPC-4
- and
bla
KPC-5
-bearing plasmids recovered from two
K. pneumoniae
strains, which were isolated from a single New Jersey hospital in 2005 and 2006, respectively. IncN plasmid pBK31551 is 84 kb in length and harbors
bla
KPC-4
,
bla
TEM-1
,
qnrB2
,
aac
(
3
)
-Ib
,
aph
(
3′
)
-I
,
qacF
,
qacE
Δ
1
,
sul1
, and
dfrA14
, which confer resistance to β-lactams, quinolones, aminoglycosides, quaternary ammonium compounds, and co-trimoxazole. The conserved regions within pBK31551 are similar to those of other IncN plasmids. Surprisingly, analysis of the Tn
4401
sequence revealed a large IS
110
- and Tn
6901
-carrying element (8.3 kb) inserted into the
istA
gene, encoding glyoxalase/bleomycin resistance, alcohol dehydrogenase, and
S
-formylglutathione hydrolase. Plasmid pBK31567 is 47 kb in length and harbors
bla
KPC-5
,
dfrA5
,
qacE
Δ
1
, and
sul1
. pBK31567 belongs to a novel IncX subgroup (IncX5) and possesses a highly syntenic plasmid backbone like other IncX plasmids; however, sequence similarity at the nucleotide level is divergent. The
bla
KPC-5
gene is carried on a Tn
4401
element and differs from the genetic environment of
bla
KPC-5
described in
Pseudomonas aeruginosa
strain P28 from Puerto Rico. This study underscores the genetic diversity of multidrug-resistant plasmids involved in the spread of
bla
KPC
genes and highlights the mobility and plasticity of Tn
4401
. Comparative genomic analysis provides new insights into the evolution and dissemination of KPC plasmids belonging to different incompatibility groups.
Clonal Structure, Extended-Spectrum β-Lactamases, and Acquired AmpC-Type Cephalosporinases of Escherichia coli Populations Colonizing Patients in Rehabilitation Centers in Four Countries
Authors: Izdebski R. , Baraniak A. , Fiett J. , Adler A. , Kazma M. , Salomon J. , Lawrence C. , Rossini A. , Salvia A. , Vidal Samso J. , Fierro J. , Paul M. , Lerman Y. , Malhotra-Kumar S. , Lammens C. , Goossens H. , Hryniewicz W. , Brun-Buisson C. , Carmeli Y. , Gniadkowski M.
Published: November 2012

The prospective project MOSAR was conducted in five rehabilitation units: the Berck Maritime Hôpital (Berck, France), Fondazione Santa Lucia (Rome, Italy), Guttmann Institute (GI; Barcelona, Spain), and Loewenstein Hospital and Tel-Aviv Souraski Medical Center (TA) (Tel-Aviv, Israel). Patients were screened for carriage of
Enterobacteriaceae
resistant to expanded-spectrum cephalosporins (ESCs) from admission until discharge. The aim of this study was to characterize the clonal structure, extended-spectrum β-lactamases (ESBLs), and acquired AmpC-like cephalosporinases in the
Escherichia coli
populations collected. A total of 376 isolates were randomly selected. The overall number of sequence types (STs) was 76, including 7 STs that grouped at least 10 isolates from at least three centers each, namely, STs 10, 38, 69, 131, 405, 410, and 648. These clones comprised 65.2% of all isolates, and ST131 alone comprised 41.2%. Of 54 STs observed only in one center, some STs played a locally significant role, like ST156 and ST393 in GI or ST372 and ST398 in TA. Among 16 new STs, five arose from evolution within the ST10 and ST131 clonal complexes. ESBLs and AmpCs accounted for 94.7% and 5.6% of the ESC-hydrolyzing β-lactamases, respectively, being dominated by the CTX-M-like enzymes (79.9%), followed by the SHV (13.5%) and CMY-2 (5.3%) types. CTX-M-15 was the most prevalent β-lactamase overall (40.6%); other ubiquitous enzymes were CTX-M-14 and CMY-2. Almost none of the common clones correlated strictly with one β-lactamase; although 58.7% of ST131 isolates produced CTX-M-15, the clone also expressed nine other enzymes. A number of clone variants with specific pulsed-field gel electrophoresis and ESBL types were spread in some locales, potentially representing newly emerging
E. coli
epidemic strains.
Effects of Etravirine on the Pharmacokinetics of the Integrase Inhibitor S/GSK1265744
Authors: Ford Susan L. , Gould Elizabeth , Chen Shuguang , Lou Yu , Dumont Etienne , Spreen William , Piscitelli Stephen
Published: November 2012

HIV integrase inhibitors such as raltegravir and elvitegravir halt HIV progression, but treatment-emergent resistance and cross-resistance have been observed. The nonnucleoside reverse transcriptase inhibitor etravirine (ETR) may be used in combination with integrase inhibitors in patients with drug resistance. This single-center, open-label, two-period, single-sequence crossover study evaluated the effects of ETR coadministration on the pharmacokinetic profile of S/GSK1265744, an investigational integrase inhibitor in phase 2 studies. Healthy subjects received 30 mg of S/GSK1265744 alone once daily for 10 days (period 1) and in combination with 200 mg of ETR twice daily for 14 days (period 2). Serial plasma samples for pharmacokinetic analyses were collected on day 10 during period 1 and on day 14 during period 2. All treatments were well tolerated. Etravirine had no effects on S/GSK1265744 geometric mean ratios of the area under the curve from time zero until the end of the dosing interval (1.01; 90% confidence interval [CI], 0.956 to 1.06), of the maximum observed plasma concentration (1.04; 90% CI, 0.987 to 1.09), or of the plasma concentration at the end of the dosing interval (0.999; 90% CI, 0.942 to 1.06). Etravirine pharmacokinetics (PK) parameters observed following coadministration with S/GSK1265744 were in the range of historical values reported for ETR alone in healthy subjects. These results indicate that 30 mg of S/GSK1265744 for 10 days as monotherapy followed by an additional 14 days in combination with ETR was well tolerated in healthy subjects and that no dose adjustment of S/GSK1265744 is required when it is coadministered with ETR.
Antimicrobial Properties of 8-Hydroxyserrulat-14-en-19-oic Acid for Treatment of Implant-Associated Infections
Authors: Nowakowska Justyna , Griesser Hans J. , Textor Marcus , Landmann Regine , Khanna Nina
Published: November 2012

Treatment options are limited for implant-associated infections (IAI) that are mainly caused by biofilm-forming staphylococci. We report here on the activity of the serrulatane compound 8-hydroxyserrulat-14-en-19-oic acid (EN4), a diterpene isolated from the Australian plant
Eremophila neglecta
. EN4 elicited antimicrobial activity toward various Gram-positive bacteria but not to Gram-negative bacteria. It showed a similar bactericidal effect against logarithmic-phase, stationary-phase, and adherent
Staphylococcus epidermidis
, as well as against methicillin-susceptible and methicillin-resistant
S. aureus
with MICs of 25 to 50 μg/ml and MBCs of 50 to 100 μg/ml. The bactericidal activity of EN4 was similar against
S. epidermidis
and its Δ
ica
mutant, which is unable to produce polysaccharide intercellular adhesin-mediated biofilm. In time-kill studies, EN4 exhibited a rapid and concentration-dependent killing of staphylococci, reducing bacterial counts by >3 log
10
CFU/ml within 5 min at concentrations of >50 μg/ml. Investigation of the mode of action of EN4 revealed membranolytic properties and a general inhibition of macromolecular biosynthesis, suggesting a multitarget activity.
In vitro
-tested cytotoxicity on eukaryotic cells was time and concentration dependent in the range of the MBCs. EN4 was then tested in a mouse tissue cage model, where it showed neither bactericidal nor cytotoxic effects, indicating an inhibition of its activity. Inhibition assays revealed that this was caused by interactions with albumin. Overall, these findings suggest that, upon structural changes, EN4 might be a promising pharmacophore for the development of new antimicrobials to treat IAI.
Biochemical Characterization of Cardiolipin Synthase Mutations Associated with Daptomycin Resistance in Enterococci
Authors: Davlieva Milya , Zhang Wanna , Arias Cesar A. , Shamoo Yousif
Published: November 2012

Daptomycin (DAP) resistance in enterococci has been linked to mutations in genes that alter the cell envelope stress response (CESR) (
liaFSR
) and changes in enzymes that directly affect phospholipid homeostasis, and these changes may alter membrane composition, such as that of cardiolipin synthase (Cls). While Cls substitutions are observed in response to DAP therapy, the effect of these mutations on Cls activity remains obscure. We have expressed, purified, and characterized Cls enzymes from both
Enterococcus faecium
S447 (residues 52 to 482; Cls447a) and
Enterococcus faecalis
S613 (residues 53 to 483; Cls613a) as well as Cls variants harboring a single-amino-acid change derived from DAP-resistant isolates of
E. faecium
.
E. faecium
Cls447a and
E. faecalis
Cls613a are tightly associated with the membrane and copurify with their substrate, phosphatidylglycerol (PG), and product, cardiolipin (CL). The amount of PG that copurifies with Cls is in molar excess to protein, suggesting that the enzyme localizes to PG-rich membrane regions. Both Cls447a
H215R
and Cls447a
R218Q
showed an increase in
V
max
(μM CL/min/μM protein) from 0.16 ± 0.01 to 0.26 ± 0.02 and 0.26 ± 0.04, respectively, indicating that mutations associated with adaptation to DAP increase Cls activity. Modeling of Cls447a to
Streptomyces
sp. phospholipase D indicates that the adaptive mutations Cls447a
H215R
and Cls447a
R218Q
are proximal to the phospholipase domain 1 (PLD1) active site and near the putative nucleophile H217. As mutations to Cls are part of a larger genomic adaptation process, increased Cls activity is likely to be highly epistatic with other changes to facilitate DAP resistance.
Detection of Francisella tularensis-Specific Antibodies in Patients with Tularemia by a Novel Competitive Enzyme-Linked Immunosorbent Assay
Authors: Sharma Neekun , Hotta Akitoyo , Yamamoto Yoshie , Fujita Osamu , Uda Akihiko , Morikawa Shigeru , Yamada Akio , Tanabayashi Kiyoshi
Published: November 2012

A novel competitive enzyme-linked immunosorbent assay (cELISA) was developed and evaluated for detection of antibodies against
Francisella tularensis
in humans. The assay is based on the ability of serum antibodies to inhibit the binding of monoclonal antibodies (MAbs) directed against
F. tularensis
lipopolysaccharide antigens. The assay was evaluated using serum samples of tularemia patients, inactivated
F. tularensis
-immunized rabbits, and
F. tularensis
-infected mice. Antibodies against
F. tularensis
were successfully detected in serum samples of tularemia patients as well as the immunized and infected animals. The cELISA method was compared to indirect ELISA (iELISA) and the commonly used microagglutination test (MA) using serum samples of 19 tularemia patients and 50 healthy individuals. The sensitivity and specificity of cELISA were 93.9 and 96.1%, respectively, in comparison to the iELISA. MA was less sensitive than cELISA with a sensitivity and specificity of only 81.8 and 98.0%, respectively. A high degree of correlation (
R
2
= 0.8226) was observed between cELISA and iELISA results. The novel cELISA developed in this study appears to be highly sensitive and specific for serodiagnosis of human tularemia. The potential of the MAb-based cELISA to be used in both human and animal samples emphasizes its usefulness for serological survey of tularemia among multiple animal species.
Large-Scale Evaluation of the Immuno-Mycologics Lateral Flow and Enzyme-Linked Immunoassays for Detection of Cryptococcal Antigen in Serum and Cerebrospinal Fluid
Authors: Hansen Jessica , Slechta E. Susan , Gates-Hollingsworth Marcellene A. , Neary Brandon , Barker Adam P. , Bauman Sean , Kozel Thomas R. , Hanson Kimberly E.
Published: November 2012

Cryptococcosis is a systemic infection caused by the pathogenic yeasts
Cryptococcus neoformans
and
C. gattii
. Detection of cryptococcal capsular antigen (CrAg) in serum and cerebrospinal fluid (CSF) plays an important diagnostic role. We prospectively compared the new Immuno-Mycologics Inc. (IMMY) lateral flow assay (LFA) and enzyme immunoassay (EIA) to our current CrAg test (Premier EIA; Meridian Bioscience Inc.). Discordant samples were retested with the latex-
Cryptococcus
antigen test (IMMY) and using serotype-specific monoclonal antibodies (MAbs). A total of 589 serum and 411 CSF specimens were tested in parallel. Qualitative agreement across assays was 97.7%. In all, 56 (41 serum and 15 CSF) samples were positive and 921 (527 serum and 394 CSF) samples were negative by all three assays. The 23 discrepant specimens were all Meridian EIA negative. Of 23 discordant specimens, 20 (87.0%) were positive by both the IMMY LFA and EIA, 2 were LFA positive only, and 1 was EIA positive only. Eleven discrepant specimens had adequate volume for latex agglutination (LA) testing; 8 were LA positive, and 3 were LA negative. LA-negative samples (2 CSF samples and 1 serum) had low IMMY LFA/EIA titers (≤1:10). Serotype-specific MAb analysis of the LA-positive samples suggested that these specimens contained CrAg epitopes similar to those of serotype C strains. In conclusion, the IMMY assays showed excellent overall concordance with the Meridian EIA. Assay performance differences were related to issues of analytic sensitivity and possible serotype bias. Incomplete access to patient-level data combined with low specimen volumes limited our ability to fully resolve discrepant results.
Population Dynamics of Tuberculous Bacilli in Cameroon as Assessed by Spoligotyping
Authors: Koro Koro Francioli , Kamdem Simo Yannick , Piam Félix Fotso , Noeske Jurgen , Gutierrez Cristina , Kuaban Christopher , Eyangoh Sara Irène
Published: November 2012

Genetic assessment by spoligotyping of 565
Mycobacterium tuberculosis
complex strains collected from the Western Region of Cameroon between 2004 and 2005 has confirmed the establishment of the “Cameroon family” as the leading cause of tuberculosis in 45.9% of cases and evidenced the rapid quasi extinction of
Mycobacterium africanum
, isolated in 3.3% of tuberculosis cases.