bluuebunny/crossref_metadata_2025 huggingface dataset:
Basic Methods for Monitoring Intracellular Ca2+ in Cardiac Myocytes Using Fluo-3
Authors: Bito Virginie , Sipido Karin R. , Macquaide Niall
Published: April 2015

In cardiac myocytes, the physiological increase of intracellular calcium, the [Ca2+]i transient, elicited during excitation–contraction coupling typically reaches a peak amplitude of up to 1 µm, from a resting value of ∼100 nm, within 50–100 msec, depending on the species. Various conditions will affect the amplitude and rise time of the [Ca2+]i transient and, depending on the nature of the Ca2+ signals under study, a variety of different probes are available for monitoring changes in intracellular Ca2+. In this protocol, we focus on Fluo-3, which exists in the cytosol in its salt form K5Fluo-3. This form is practically nonfluorescent in the absence of Ca2+, but the fluorescence increases dramatically on Ca2+ binding. Although Fluo-3 is a single excitation–emission dye, it has a number of advantages for investigators, including an ideal dissociation constant (Kd) value and high quantum yield, meaning that it can be used at low concentrations that introduce minimal buffering. Here, we describe the basic setup and methodology for recording the global cytosolic [Ca2+]i transient with this probe during simultaneous patch-clamp and whole-cell recording of membrane voltage or of ionic currents under voltage clamp.
Culturing Adult Stem Cells from Mouse Small Intestinal Crypts
Authors: Hamilton Kathryn E. , Crissey Mary Ann S. , Lynch John P. , Rustgi Anil K.
Published: April 2015

In recent years, the study of primary cells in culture has evolved from an extraphysiological, two-dimensional platform to novel, three-dimensional platforms in which the addition of matrix components and/or supporting cells provide an ex vivo niche. Such studies have provided the basis on which to study more advanced physiological processes in detail, including multilayered, long-term cultures, epithelial–stromal interactions, and stem cell behaviors that more closely recapitulate normal morphology than two-dimensional culture. Various techniques for three-dimensional organotypic culture and crypt culture of primary cells from mouse and human small intestine and colon have been described. These methods have allowed for the study of specific stem cell characteristics, including survival, self-renewal, and long-term growth in culture, as well as the ability to propagate all the appropriate progenitor and postmitotic lineages. These assays have become a widely accepted functional measure of “stemness” and, in combination with lineage-tracing experiments in various genetically engineered mouse models, have been critical in the identification of specific markers of intestinal stem cells. In this protocol we draw upon recently described methods for the isolation and culture of mouse small intestinal enterospheres/enteroids from isolated crypts and/or single cells. Cultures of murine colon epithelium, as well as human small intestine and colon, require additional growth factors not discussed here. The description provided here represents current knowledge, and it is possible, if not likely, that modifications in the future will emerge.
Three-Dimensional Organotypic Culture of Stratified Epithelia
Authors: Rhim Andrew D. , Rustgi Anil K.
Published: April 2015

One of the limitations of conventional tissue culture on flat two-dimensional surfaces is the loss of complex interactions between the epithelium and stroma. We have devised a culture system that recreates the salient features of the stratified epithelium using primary cell cultures from mouse models. The protocol described here is applicable to the esophageal epithelium, but stratified epithelial cells from other organs (e.g., skin) can be grown. Once established, the system can be used to interrogate the effect of various pharmacologic and genetic manipulations on epithelial homeostasis and invasion.
Arabidopsis BIRD Zinc Finger Proteins Jointly Stabilize Tissue Boundaries by Confining the Cell Fate Regulator SHORT-ROOT and Contributing to Fate Specification
Authors: Long Yuchen , Smet Wouter , Cruz-Ramírez Alfredo , Castelijns Bas , de Jonge Wim , Mähönen Ari Pekka , Bouchet Benjamin P. , Perez Gabino Sanchez , Akhmanova Anna , Scheres Ben , Blilou Ikram
Published: April 2015

Plant cells cannot rearrange their positions; therefore, sharp tissue boundaries must be accurately programmed. Movement of the cell fate regulator SHORT-ROOT from the stele to the ground tissue has been associated with transferring positional information across tissue boundaries. The zinc finger BIRD protein JACKDAW has been shown to constrain SHORT-ROOT movement to a single layer, and other BIRD family proteins were postulated to counteract JACKDAW's role in restricting SHORT-ROOT action range. Here, we report that regulation of SHORT-ROOT movement requires additional BIRD proteins whose action is critical for the establishment and maintenance of the boundary between stele and ground tissue. We show that BIRD proteins act in concert and not in opposition. The exploitation of asymmetric redundancies allows the separation of two BIRD functions: constraining SHORT-ROOT spread through nuclear retention and transcriptional regulation of key downstream SHORT-ROOT targets, including SCARECROW and CYCLIND6. Our data indicate that BIRD proteins promote formative divisions and tissue specification in the Arabidopsis thaliana root meristem ground tissue by tethering and regulating transcriptional competence of SHORT-ROOT complexes. As a result, a tissue boundary is not “locked in” after initial patterning like in many animal systems, but possesses considerable developmental plasticity due to continuous reliance on mobile transcription factors.
X-rays only when you want them: optimized pump–probe experiments using pseudo-single-bunch operation
Authors: Hertlein M. P. , Scholl A. , Cordones A. A. , Lee J. H. , Engelhorn K. , Glover T. E. , Barbrel B. , Sun C. , Steier C. , Portmann G. , Robin D. S.
Published: April 2015

Laser pump–X-ray probe experiments require control over the X-ray pulse pattern and timing. Here, the first use of pseudo-single-bunch mode at the Advanced Light Source in picosecond time-resolved X-ray absorption experiments on solutions and solids is reported. In this mode the X-ray repetition rate is fully adjustable from single shot to 500 kHz, allowing it to be matched to typical laser excitation pulse rates. Suppressing undesired X-ray pulses considerably reduces detector noise and improves signal to noise in time-resolved experiments. In addition, dose-induced sample damage is considerably reduced, easing experimental setup and allowing the investigation of less robust samples. Single-shot X-ray exposures of a streak camera detector using a conventional non-gated charge-coupled device (CCD) camera are also demonstrated.
Synthesis and structural elucidation of new complexes of 2,4- and 3,5-dimethyl diphenyldithiophosphates with cobalt(II)
Authors: Kumar Sandeep , Khajuria Ruchi , Jassal Amanpreet Kaur , Hundal Maninder S. , Pandey Sushil K.
Published: April 2015

A new series of donor-stabilized addition complexes of cobalt(II) with disubstituted diphenyldithiophosphates [{(ArO)2PS2}2CoL2] {Ar = 2,4-(CH3)2C6H3[(1)–(3)] and 3,5-(CH3)2C6H3[(4)–(6)];L= C5H5N [(1), (4)], 3,4-(CH3)2C5H3N [(2), (5)] and 4-(C2H5)C5H4N [(3), (6)]} were successfully isolated and characterized by elemental analyses, magnetic moment measurements, IR and single-crystal X-ray analysis. Complexes (3), (4) and (5) crystallize in the monoclinic space groupsP21/n,P21/nandP21/c, respectively, whereas complex (6) crystallizes in the triclinic space group P\bar 1. X-ray diffraction analysis of complexes (3)–(6) reveals a six-coordinated distorted octahedral geometry for the CoS4N2chromophore defined by two chelating diphenyldithiophosphate anions as well as two N-donor ligands. Two diphenyldithiophosphate ligands are coordinated to the cobalt ion as a bidentate ligand chelatingviathe two thiolate S atoms. Each of them forms a four-membered chelate ring in the equatorial plane. The N atoms from two donor ligands are axially coordinated to the Co atom in a mutuallytransposition.
Structure of the mixed-metal carbonate KAgCO3revisited: order–disorder (OD) polytypism and allotwinning
Authors: Hans Philipp , Stöger Berthold , Weil Matthias , Zobetz Erich
Published: April 2015

Crystals of KAgCO3belong to an order–disorder (OD) family of structures composed of layers of two kinds. There are two polytypes with a maximum degree of order [MDO1:Pccb; MDO2:Ibca, doubleda-axis compared with MDO1], which are both realised to a different extent in two crystals under investigation [volume fraction MDO1:MDO2in crystal (I): 0.0216:0.9784 (3) and in crystal (II): 0.9657:0.0343 (3)]. Sharp diffraction spots and the absence of diffuse scattering indicate highly ordered macroscopic domains. The structure of KAgCO3was refined concurrently against all reflections using an allotwin model (addition of the intensities of both domains). It is shown that a disorder model refined against reflections of only one domain can lead to a significant overestimation of the volume fraction of this domain.
Crystal structure of (1,3-di-tert-butyl-η5-cyclopentadienyl)trimethylhafnium(IV)
Authors: Pérez-Redondo Adrián , Varela-Izquierdo Víctor , Yélamos Carlos
Published: April 2015

The molecule of the title organometallic hafnium(IV) compound, [Hf(CH3)3(C13H21)] or [HfMe3(η5-C5H3-1,3-tBu2)], adopts the classical three-legged piano-stool geometry for monocyclopentadienylhafnium(IV) derivatives with the three methyl groups bonded to the Hf(IV) atom at the legs. The C atoms of the twotert-butyl group bonded to the cyclopentadienyl (Cp) ring are 0.132 (5) and 0.154 (6) Å above the Cp least-squares plane. There are no significant intermolecular interactions present between the molecules in the crystal structure.
Crystal structure of bis[N-phenyl-2-(1,2,3,4-tetrahydronaphthalen-1-ylidene)hydrazinecarbothioamidato-κ2N2,S]zinc dimethyl sulfoxide monosolvate
Authors: Cruz Santana Genelane , Gimenez Iara de Fátima , Näther Christian , Jess Inke , de Oliveira Adriano Bof
Published: April 2015

The reaction of theN-phenyl-2-(1,2,3,4-tetrahydronaphthalen-1-ylidene)hydrazinecarbothioamide ligand with zinc acetate dihydrate in a 2:1 molar ratio yielded a yellow solid, which was crystallized from DMSO to obtain the title compound, [Zn(C17H16N3S)2]·C2H6OS. The ZnIIion is four-coordinated in a distorted tetrahedral environment by two deprotonated ligands. Each ligand acts as anN,S-donor, forming a five-membered metallacycle. The maximum deviation from the mean plane of the N–N–C–S chelate group is 0.0029 (14) Å for theN-donor atom of one ligand and 0.0044 (14) Å for the non-coordinating N atom of the second. The dihedral angle between the planes of the two chelate groups is 72.80 (07)°. Bond lengths in the ligands are compared with those in the crystal structure of the free ligand. In the crystal, complex molecules are connected by dimethyl sulfoxide solvate moleculesviaN—H...O hydrogen-bonding interactions, building a one-dimensional hydrogen-bonded polymer along thea-axis direction. The S atom and one C atom of the dimethyl sulfoxide solvate molecules are disordered over two sets of sites with an occupancy ratio of 0.6:0.4.
Crystal structure of [1-(3-chlorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-4-yl](p-tolyl)methanone
Authors: Kumar Balbir , Nakum Kiran J. , Jadeja R. N. , Kant Rajni , Gupta Vivek K.
Published: April 2015

In the title compound C18H15ClN2O2, the dihedral angles between the central pyrazole ring and the pendant chlorobenzene andp-tolyl rings are 17.68 (10) and 51.26 (12)°, respectively. An intramolecular O—H...O hydrogen bond is observed, which closes anS(6) ring.
Crystal structure of methyl 1-methyl-2-oxospiro[indoline-3,2′-oxirane]-3′-carboxylate
Authors: Savithri M. P. , Yuvaraj P. S. , Reddy B. S. R. , Raja R. , SubbiahPandi A.
Published: April 2015

In the title compound, C12H11NO4, the dihedral angle between the indole ring system (r.m.s. deviation = 0.019 Å) and the oxirane ring is 88.8 (2)°. The oxirane O atom and the bridging ester O atom are in an approximate syn conformation [O—C—C—O = −25.4 (3)°] In the crystal, inversion dimers linked by pair of C—H...O hydrogen bonds generateR22(8) loops, where the C—H donor group forms part of the oxirane ring. A second C—H...O interaction arising from one of the C—H groups of the benzene ring links the dimers into [001] double chains.
Crystal structure of (2-methyl-4-phenyl-4H-benzo[4,5]thiazolo[3,2-a]pyrimidin-3-yl)(phenyl)methanone
Authors: Sankar T. , Naveen S. , Lokanath N. K. , Gunasekaran K.
Published: April 2015

In the title compound, C24H18N2OS, the pyrimidine ring has a flat envelope conformation with the methine C atom as the flap. The attached phenyl and benzoyl rings are inclined to the mean plane of the pyrimidine ring by 84.87 (8) and 75.33 (9)°, respectively. The benzothiazolo group is planar (r.m.s. deviation = 0.009 Å) and inclined to the mean plane of the pyrimidine ring by 3.27 (6)°. In the crystal, molecules are linked by pairs of C—H...N hydrogen bonds, forming inversion dimers.
The engagement continuum model using corporate social responsibility as an intervention for sustained employee engagement
Authors: Valentin Marie Anttonitte , Valentin Celestino C , Nafukho Fredrick Muyia
Published: April 2015

Purpose

– The purpose of this paper is to explore implications of motivational potential that are highly correlated to the self-determination theory (SDT) (intrinsic motivating factors), in relation to corporate social responsibility (CSR). This paper specifies key antecedents of engagement within the theoretical framework of the self-determination theory as it relates to employee engagement and CSR.

Design/methodology/approach

– The methods used for the purpose of this paper include a review of the relevant literature utilizing the descriptors of employee engagement, SDT and CSR. Alternative descriptors were not queried. The authors then selected articles that were found to be most cited, reviewed such articles and began to analyze the literature, synthesize and formulate connections.

Findings

– Based on research findings, a conceptual model was formulated and posited for research and practice. It is demonstrated in the paper that employee engagement has a wide range of benefits for all involved and focuses on key antecedents of engagement created through CSR initiatives and intrinsic motivating factors as pointed out from SDT, which may serve to provide a comprehensive representation of the likely influences of intrinsic motivating drivers on employee engagement.

Research limitations/implications

– The main limitations of this paper is that it is conceptual in nature and, hence, the need for a study designed to empirically test the conceptual model developed in this research.

Originality/value

– The result and contribution to the field of human resource development is the development of the engagement continuum model from which employee engagement emerges through the dynamic interplay of CSR as an intervention, creating positive results using the theoretical framework of SDT and resulting in a perceived sustained state of employee engagement.
Professional development of HR practitioners – a phenomenographic study
Authors: Bailey Moira
Published: April 2015

Purpose

– The purpose of this paper is an investigation into the experiences of professional development of human resource (HR) practitioners in the North of Scotland, and the use of non-formal learning in that development.

Design/methodology/approach

– In-depth semi-structured interviews from a purposively selected sample of HR practitioners were conducted. Data from these interviews was analysed on a phenomenographic basis, to discover the qualitatively different ways in which HR practitioners describe, experience, understand and analyse their professional development and the use of non-formal learning in that development.

Findings

– What emerged from the analysis were two sets of categories of description, one for each of the phenomena. An outcome space for each of the phenomena emerged, illustrating the hierarchical relationship within each set of categories of description as well as the dimensions of variation relating to the phenomena. These outcome spaces represent the collective experience of the practitioners on the subjects of professional development and non-formal learning.

Research limitations/implications

– Sample size and the specific geographical area are acknowledged as limitations. Another factor which may be considered a limitation is that the author's position as an HR lecturer with a keen interest in the subject could lead to this being considered an “insider” study. All these factors are acknowledged. These have been mitigated against by the careful preparation undertaken during the research process which resulted from the author's awareness of these limitations.

Originality/value

– This study has given a voice to the HR practitioners in the North of Scotland with regard to their experiences and attitudes towards their professional development and the role of non-formal learning in that development. This study gives employers, other practitioners and professional bodies an opportunity to learn from the practitioners themselves as to how they can help practitioners in terms of their development.
Energy expenditure and body size are targets of natural selection across a wide geographic range, in a terrestrial invertebrate
Authors: Bartheld José Luis , Gaitán‐Espitia Juan Diego , Artacho Paulina , Salgado‐Luarte Cristian , Gianoli Ernesto , Nespolo Roberto F.
Published: April 2015

One of the central questions in evolutionary ecology is how different functional capacities impact fitness, and how it varies across populations. For instance, do phenotypic attributes influence fitness similarly across geographic gradients? Which traits (physiological, morphological and life history) are most likely to be targets of natural selection? Do particular combinations of traits maximize fitness?

In a semi‐natural experiment, we analysed introduced populations of an invasive species, the garden snail (Cornu aspersum) in Chile, which show low levels of genetic differentiation in spite of the distance. Specifically, we addressed whether the magnitude, sign and form of selection in snail populations could explain the differentiation (or its absence) among populations.

A common garden/reciprocal transplant experiment was performed in three populations (La Serena, Constitución and Valdivia) that span a 1300‐km latitudinal gradient and differ markedly in climate (semi‐arid north to humid south). We released c. 450 individuals per population (two generations after field‐captured snails) in replicated enclosures at the range extremes (La Serena and Valdivia). Morphological (size and shell darkness), physiological (standard metabolic rate and digestive efficiency) and life‐history [growth rate (GR)] traits were measured in all snails before the release. Survival was recorded monthly during 1 year.

Survival was significantly higher in snails from La Serena than in snails from Constitución and Valdivia, when raised at La Serena. However, at Valdivia, survival was not different among source populations. Interestingly, we found negative correlational selection in MB and SMRR at La Serena, whereas at Valdivia we only found directional selection on GR and MB, and stabilizing selection on SMRR.

These results suggest that selection on physiological traits related with energy allocation is pervasive, irrespective of climate and distance.
Investigating the Postmortem Molecular Biology of Cartilage and its Potential Forensic Applications
Authors: Bolton Shawna N. , Whitehead Michael P. , Dudhia Jayesh , Baldwin Timothy C. , Sutton Raul
Published: April 2015

This study investigated the postmortem molecular changes that articular cartilage undergoes following burial. Fresh pig trotters were interred in 30‐cm‐deep graves at two distinct locations exhibiting dissimilar soil environments for up to 42 days. Extracts of the metacarpophalangeal (MCP) and metatarsophalangeal (MTP) joint cartilage from trotters disinterred weekly over 6 weeks were analyzed by Western blot against the monoclonal antibody 2‐B‐6 to assess aggrecan degradation. In both soil conditions, aggrecan degradation by‐products of decreasing molecular size and complexity were observed up to 21 days postmortem. Degradation products were undetected after this time and coincided withMCP/MTPjoint exposure to the soil environment. These results show that cartilage proteoglycans undergo an ordered molecular breakdown, the analysis of which may have forensic applications. This model may prove useful for use as a human model and for forensic investigations concerning crimes against animals and the mortality of endangered species.
A Noninvasive and Speculative Method of Visualizing Latent Fingerprint Deposits on Thermal Paper
Authors: Bond John W.
Published: April 2015

Latent fingerprint deposits on thermal paper have been visualized noninvasively at visible wavelengths when illuminated with aUV‐A light source (peak 365 nm). A higher intensityUVsource (250 W/m2at 0.38 m) gave superior fingerprint visibility when compared with a 60 W/m2(at 0.4 m) source. Removing the visible (blue) component of the light source emission did not adversely affect the visibility of the fingerprint. Sample fingerprints from 100 donors, when examined 24 h after deposition, produced identifiable fingerprints from nearly 34% of fingerprint deposits. A mechanism for the observed visibility is proposed based on low emission of visible wavelengths from areas of thermal paper coincident with the fingerprint deposit, when illuminated withUV. This is likely due to a weak color change in the thermal paper dye arising from protonated amino acid components of the sweat. This effect was not observed on nonthermal paper.
Do microsporidia function as “biological weapon” for Harmonia axyridis under natural conditions?
Authors: Gegner Tobias , Otti Oliver , Tragust Simon , Feldhaar Heike
Published: April 2015

Invasive alien species, such as the multicoloured Asian ladybird Harmonia axyridis, are often regarded as major drivers of biodiversity loss. Therefore understanding which characteristics or mechanisms contribute to their invasive success is important. Here the role of symbiotic microsporidia in the hemolymph of H. axyridis was investigated in the context of intraguild predation between wild‐caught H. axyridis and the native ladybird species Coccinella septempunctata. The microsporidia were recently discussed to contribute to the unpalatability of Harmonia for other coccinellids during intraguild predation and to function as “biological weapons”. In the present study, visual detection of microsporidia in hemolymph samples revealed that 73.5 % of H. axyridis were infected. Intraguild predation experiments between larvae of the two species showed a significant competitive advantage for H. axyridis, even against larger larvae of C. septempunctata. Adult C. septempunctata always killed and fed on H. axyridis larvae. However only 11.4 % (4 of 47) of C. septempunctata that fed on infected H. axyridis died within 4 months. In contrast to previous studies this suggests that microsporidia or harmonine, the chemical defense compound of H. axyridis, do not lead to death of C. septempunctata preying on larvae of H. axyridis. Instead our results support the idea that competitive advantage during intraguild predation greatly facilitates the success of H. axyridis and that this may help this highly invasive species to outcompete native species. The impact of microsporidia on Harmonia itself as well as on interspecific interactions require further studies.
The utility of three‐dimensional optical projection tomography in nerve injection injury imaging
Authors: Cvetko E. , Čapek M. , Damjanovska M. , Reina M. A. , Eržen I. , Stopar‐Pintarič T.
Published: April 2015

The examination of nerve microarchitecture has hitherto been limited solely to two‐dimensional imaging techniques. The objective of this study was to evaluate the ability of optical projection tomography to discern the nerve microarchitecture and injection injury in three dimensions. Five piglets were studied, whose median and lingual nerves were unilaterally injected post mortem with preset volumes of local anaesthetic, excised and subsequently made transparent with benzyl alcohol benzyl benzoate. Images were captured in three dimensions. The same contralateral nerves were used as controls. Using optical projection tomography, we observed differences between the internal organisation of the median and the lingual nerves, which potentially explain the variations in their susceptibility to injury. This was demonstrated in three dimensions as a disruption to the fascicles in the lingual nerve, and their displacement in the median nerve. This new technology offers potential for studying nerve microarchitecture topography and its tolerance to injection injury.
The Nordic Obstetric Surveillance Study: a study of complete uterine rupture, abnormally invasive placenta, peripartum hysterectomy, and severe blood loss at delivery
Authors: Colmorn Lotte B. , Petersen Kathrine B. , Jakobsson Maija , Lindqvist Pelle G. , Klungsoyr Kari , Källen Karin , Bjarnadottir Ragnheidur I. , Tapper Anna‐Maija , Børdahl Per E. , Gottvall Karin , Thurn Lars , Gissler Mika , Krebs Lone , Langhoff‐Roos Jens
Published: April 2015

Objective

To assess the rates and characteristics of women with complete uterine rupture, abnormally invasive placenta, peripartum hysterectomy, and severe blood loss at delivery in the Nordic countries.

Design

Prospective, Nordic collaboration.

Setting

The Nordic Obstetric Surveillance Study (NOSS) collected cases of severe obstetric complications in the Nordic countries from April 2009 to August 2012.

Sample and methods

Cases were reported by clinicians at the Nordic maternity units and retrieved from medical birth registers, hospital discharge registers, and transfusion databases by using International Classification of Diseases, 10th revision codes on diagnoses and the Nordic Medico‐Statistical Committee Classification of Surgical Procedure codes.

Main outcome measures

Rates of the studied complications and possible risk factors among parturients in the Nordic countries.

Results

The studied complications were reported in 1019 instances among 605 362 deliveries during the study period. The reported rate of severe blood loss at delivery was 11.6/10 000 deliveries, complete uterine rupture was 5.6/10 000 deliveries, abnormally invasive placenta was 4.6/10 000 deliveries, and peripartum hysterectomy was 3.5/10 000 deliveries. Of the women, 25% had two or more complications. Women with complications were more often >35 years old, overweight, with a higher parity, and a history of cesarean delivery compared with the total population.

Conclusion

The studied obstetric complications are rare. Uniform definitions and valid reporting are essential for international comparisons. The main risk factors include previous cesarean section. The detailed information collected in the NOSS database provides a basis for epidemiologic studies, audits, and educational activities.
Efficacy of tenofovir switch therapy for nucleos(t)ide‐experienced patients with chronic hepatitis B
Authors: Lo A. O.‐S. , Wong V. W.‐S. , Wong G. L.‐H. , Tse Y.‐K. , Chan H.‐Y. , Chan H. L.‐Y.
Published: April 2015

Background

Tenofovir disoproxil fumarate has been used in chronic hepatitis B patients with suboptimal virologic response to nucleos(t)ide analogues. The efficacy of tenofovir switch therapy has not been well studied in Asian patients.

Aim

To evaluate the efficacy of tenofovir switch therapy in nucleos(t)ide‐experienced patients, and identify the factors associated with treatment response of tenofovir switch therapy.

Methods

Nucleos(t)ide‐experienced hepatitis B e antigen‐positive and ‐negative patients prescribed with tenofovir were retrospectively identified and recruited for prospective analysis. Hepatitis B virus (HBV) DNA and other biochemical parameters were monitored in regular 3–6 monthly follow‐up visits. Primary efficacy endpoint was maintained‐virologic response with tenofovir switch therapy, defined as undetectable HBV DNA (<20 IU/mL) until the last follow‐up visit.

Results

An overall of 214/252 (84.9%) patients achieved maintained‐virologic response after 22 (7–55) months of tenofovir switch therapy. On multivariate analysis, a lower HBV DNA level at the time of switching to tenofovir was an independent factor associated with treatment efficacy. Maintained‐virologic response after switching to tenofovir was achieved in 177/190 (93.2%) patients with HBV DNA <20 000 IU/mL vs. 37/62 (59.7%) patients with HBV DNA ≥20 000 IU/mL (P < 0.001). Absence of genotypic resistance to lamivudine or adefovir dipivoxil was not associated with improved treatment outcome.

Conclusions

Tenofovir switch therapy is an effective treatment strategy in nucleos(t)ide‐experienced chronic hepatitis B patients. However, in patients with HBV DNA ≥20 000 IU/mL at the time of switching to tenofovir, the chance of achieving maintained undetectable HBV DNA is significantly reduced.
Population pharmacokinetic analysis of tacrolimus in Mexican paediatric renal transplant patients: role of CYP3A5 genotype and formulation
Authors: Jacobo‐Cabral Carlos Orlando , García‐Roca Pilar , Romero‐Tejeda Elba Margarita , Reyes Herlinda , Medeiros Mara , Castañeda‐Hernández Gilberto , Trocóniz Iñaki F.
Published: April 2015

Aims

The aims of this study were (i) to develop a population pharmacokinetic (PK) model of tacrolimus in a Mexican renal transplant paediatric population (n = 53) and (ii) to test the influence of different covariates on its PK properties to facilitate dose individualization.

Methods

Population PK and variability parameters were estimated from whole blood drug concentration profiles obtained at steady‐state using the non‐linear mixed effect modelling software NONMEM® Version 7.2.

Results

Tacrolimus PK profiles exhibited high inter‐patient variability (IPV). A two compartment model with first order input and elimination described the tacrolimus PK profiles in the studied population. The relationship between CYP3A5 genotype and tacrolimus CL/F was included in the final model. CL/F in CYP3A5*1/*1 and *1/*3 carriers was approximately 2‐ and 1.5‐fold higher than in CYP3A5*3/*3 carriers (non‐expressers), respectively, and explained almost the entire IPV in CL/F. Other covariates retained in the final model were the tacrolimus dose and formulation type. Limustin® showed markedly lower concentrations than the rest of the formulations.

Conclusions

Population PK modelling of tacrolimus in paediatric renal transplant recipients identified the tacrolimus formulation type as a significant covariate affecting the blood concentrations and confirmed the previously reported significant effect of CYP3A5 genotype on CL/F. It allowed the design of a proposed dosage based on the final model that is expected to help to improve tacrolimus dosing.
Statistical monitoring of the hand, foot and mouth disease in China
Authors: Zhang Jingnan , Kang Yicheng , Yang Yang , Qiu Peihua
Published: April 2015

In a period starting around 2007, the Hand, Foot, and Mouth Disease (HFMD) became wide‐spreading in China, and the Chinese public health was seriously threatened. To prevent the outbreak of infectious diseases like HFMD, effective disease surveillance systems would be especially helpful to give signals of disease outbreaks as early as possible. Statistical process control (SPC) charts provide a major statistical tool in industrial quality control for detecting product defectives in a timely manner. In recent years, SPC charts have been used for disease surveillance. However, disease surveillance data often have much more complicated structures, compared to the data collected from industrial production lines. Major challenges, including lack of in‐control data, complex seasonal effects, and spatio‐temporal correlations, make the surveillance data difficult to handle. In this article, we propose a three‐step procedure for analyzing disease surveillance data, and our procedure is demonstrated using the HFMD data collected during 2008–2009 in China. Our method uses nonparametric longitudinal data and time series analysis methods to eliminate the possible impact of seasonality and temporal correlation before the disease incidence data are sequentially monitored by a SPC chart. At both national and provincial levels, our proposed method can effectively detect the increasing trend of disease incidence rate before the disease becomes wide‐spreading.
Effects of chronic treatment with the new ultra‐long‐acting β2‐adrenoceptor agonist indacaterol alone or in combination with the β1‐adrenoceptor blocker metoprolol on cardiac remodelling
Authors: Rinaldi Barbara , Donniacuo Maria , Sodano Loredana , Gritti Giulia , Martuscelli Eugenio , Orlandi Augusto , Rafaniello Concetta , Rossi Francesco , Calzetta Luigino , Capuano Annalisa , Matera Maria Gabriella
Published: April 2015

Background and Purpose

The ability of a chronic treatment with indacaterol, a new ultra‐long‐acting β2‐adrenoceptor agonist, to reverse cardiac remodelling and its effects in combination with metoprolol, a selective β1‐adrenoceptor antagonist, were investigated on myocardial infarction in a rat model of heart failure (HF).

Experimental Approach

We investigated the effects of indacaterol and metoprolol, administered alone or in combination, on myocardial histology, β‐adrenoceptor‐mediated pathways, markers of remodelling and haemodynamic parameters in a rat model of HF. Five groups of rats were assessed: sham‐operated rats; HF rats; HF + indacaterol 0.3 mg·kg−1·day−1; HF + metoprolol 100 mg·kg−1·day−1; HF + metoprolol + indacaterol. All pharmacological treatments continued for 15 weeks.

Key Results

Treatment with either indacaterol or metoprolol significantly reduced the infarct size in HF rats. However, the combination of indacaterol and metoprolol reduced the infarct size even further, reduced both BP and heart rate, reversed the decrease in ejection fraction, normalized left ventricular systolic and diastolic internal diameters, normalized the decreased β1 adrenoceptor mRNA expression as well as cardiac cAMP levels and reduced cardiac GPCR kinase 2 expression, compared with the untreated HF group.

Conclusion and Implications

The results of our study demonstrated an additive interaction between indacaterol and metoprolol in normalizing and reversing cardiac remodelling in our experimental model of HF. The translation of these findings to clinical practice might be of interest, as this combination of drugs could be safer and more effective in patients suffering from HF and COPD.
Synergistic enhancement of parasiticidal activity of amphotericin B using copaiba oil in nanoemulsified carrier for oral delivery: an approach for non‐toxic chemotherapy
Authors: Gupta Pramod K , Jaiswal Anil K , Asthana Shalini , Teja B Venkatesh , Shukla Prashant , Shukla Minakshi , Sagar Neeti , Dube Anuradha , Rath Srikanta K , Mishra Prabhat R
Published: April 2015

Background and Purpose

The aim of this study was to devise a nanoemulsified carrier system (CopNEC) to improve the oral delivery of amphotericin B (AmB) by increasing its oral bioavailability and synergistically enhance its antileishmanial activity with copaiba oil (Cop).

Experimental Approach

The AmB encapsulated NEC (CopNEC‐AmB) comprised of Cop, d‐α‐tocopheryl polyethylene glycol 1000 succinate and phosphatidylcholine was prepared by high‐pressure homogenization method. Stability study of CopNEC‐AmB was carried out in simulated gastric fluid and simulated intestinal fluid. The CopNEC‐AmB and plain AmB were compared as regards their in vitro antileishmanial activity, pharmacokinetics, organ distribution and toxicity.

Key Results

The optimal CopNEC‐AmB had a small globule size, low polydispersity index, high ζ potential and encapsulation efficiency. The high resolution transmission electron microscopy illustrated spherical particle geometry with homogeny in their sizes. The optimal CopNEC‐AmB was found to be stable in gastrointestinal fluids showing insignificant changes in globule size and encapsulation efficiency. The AUC0–48 value of CopNEC‐AmB in rats was significantly improved showing 7.2‐fold higher oral bioavailability than free drug. The in vitro antileishmanial activity of CopNEC‐AmB was significantly higher than that of the free drug as Cop synergistically enhanced the antileishmanial effect of AmB by causing drastic changes in the morphology of Leishmania parasite and rupturing its plasma membrane. The CopNEC‐AmB showed significantly less haemolytic toxicity and cytotoxicity and did not change the histopathology of kidney tissues as compared with AmB alone.

Conclusions and Implications

This prototype CopNEC formulation showed improved bioavailability and had a non‐toxic synergistic effect on the antileishmanial activity of AmB.
Discovery and pharmacological characterization of a novel potent inhibitor of diacylglycerol‐sensitive TRPC cation channels
Authors: Maier T , Follmann M , Hessler G , Kleemann H‐W , Hachtel S , Fuchs B , Weissmann N , Linz W , Schmidt T , Löhn M , Schroeter K , Wang L , Rütten H , Strübing C
Published: April 2015

Background and Purpose

The cation channel transient receptor potential canonical (TRPC) 6 has been associated with several pathologies including focal segmental glomerulosclerosis, pulmonary hypertension and ischaemia reperfusion‐induced lung oedema. We set out to discover novel inhibitors of TRPC6 channels and investigate the therapeutic potential of these agents.

Experimental Approach

A library of potential TRPC channel inhibitors was designed and synthesized. Activity of the compounds was assessed by measuring intracellular Ca2+ levels. The lead compound SAR7334 was further characterized by whole‐cell patch‐clamp techniques. The effects of SAR7334 on acute hypoxic pulmonary vasoconstriction (HPV) and systemic BP were investigated.

Key Results

SAR7334 inhibited TRPC6, TRPC3 and TRPC7‐mediated Ca2+ influx into cells with IC50s of 9.5, 282 and 226 nM, whereas TRPC4 and TRPC5‐mediated Ca2+ entry was not affected. Patch‐clamp experiments confirmed that the compound blocked TRPC6 currents with an IC50 of 7.9 nM. Furthermore, SAR7334 suppressed TRPC6‐dependent acute HPV in isolated perfused lungs from mice. Pharmacokinetic studies of SAR7334 demonstrated that the compound was suitable for chronic oral administration. In an initial short‐term study, SAR7334 did not change mean arterial pressure in spontaneously hypertensive rats (SHR).

Conclusions and Implications

Our results confirm the role of TRPC6 channels in hypoxic pulmonary vasoregulation and indicate that these channels are unlikely to play a major role in BP regulation in SHR. SAR7334 is a novel, highly potent and bioavailable inhibitor of TRPC6 channels that opens new opportunities for the investigation of TRPC channel function in vivo.
Anticancer Properties of Novel 4‐methylene‐1,2‐diphenylpyrazolidin‐3‐ones
Authors: Gach Katarzyna , Szymański Jacek , Pomorska Dorota , Długosz Angelika , Modranka Jakub , Michalak Marlena , Janecki Tomasz , Janecka Anna
Published: April 2015

The limited success of the currently used antitumor therapies is the driving force for organic chemists to seek new lead structures with anticancer potential. Two α‐methylene‐γ‐lactams with an additional nitrogen atom in the lactam ring, 5‐vinyl‐1,2‐diphenyl‐4‐methylenepyrazolidin‐3‐one (2a) and 5‐phenyl‐1,2‐diphenyl‐4‐methylenepyrazolidin‐3‐one (2b) have been synthesized. Their anticancer activity was assessed in MCF‐7 cells. Both compounds inhibited cell proliferation and induced DNA damage and apoptosis, with 2a being the more potent analog. Synergistic effects of 2a used in combination with known anticancer drugs, 5‐fluorouracil, taxol, and oxaliplatin were evaluated. Compound 2a significantly enhanced the antitumor action of oxaliplatin and 5‐fluorouracil, but not taxol.
Still Searching for the Zipperump‐a‐Zoo: A Reflection After 40 Years
Authors: Sternberg Robert J.
Published: April 2015

In this article, I describe chronologically my attempts over a 40‐year career to understand the nature of human intelligence. I explain how later attempts built on earlier ones, with each attempt revealing the earlier one to be too limited and narrow in the questions it asked. In my early work, I envisioned intelligence in terms of components of information processing. Later, I viewed these components as contributing to three distinct but related aspects of intelligence: analytical, creative, and practical. I came to realize the importance of contextual factors in determining what constitutes adaptive behavior. Still later, I viewed wisdom as part of the mix. The search has been rewarding, except for the fact that I have not yet completed it and never will.
Estimating the Potential Benefits of New Health Claims in Canada: The Case of Soluble Fiber and Soy Protein
Authors: Malla Stavroula , Hobbs Jill E. , Sogah Eric K.
Published: April 2015

Growing awareness of the link between diet and health has spurred growth in the functional food sector. Health Canada regulates allowable health claims on food products, and in recent years has approved health claims linking the consumption of soluble fiber from barley (2012) and psyllium (2011) to reduced/lower low‐density lipoprotein (LDL)‐cholesterol levels, a major risk factor for heart disease. A health claim linking consumption of soy protein to reduced risk of coronary heart disease (CHD) is still under consideration. Using a cost‐of‐illness approach, this paper estimates the potential economic benefits of allowing health claims for soluble fiber and soy protein in terms of reductions in the direct and indirect costs of CHD. Parameters for the economic analysis are drawn from a meta‐analysis of scientific studies examining the effect of soluble fiber and soy protein on LDL‐cholesterol levels, as well as other scientific literature. While a barley soluble fiber health claim yields nontrivial benefits in a base case scenario equal to CAD$105 million annually and ranging from $42 million to $238 million in low and high scenarios, the potential benefits of a soy protein health claim appear to be several magnitudes larger at $549 million annually in the base case and ranging from $220 million to $1.25 billion in low/high scenarios. Given the relatively slow regulatory approval process for new health claims, there may be value in using economic estimates of potential gains to help prioritize health claims approval processes.
Designing a new model for cosmetic adverse reaction monitoring in China
Authors: Lu Changfei , Cheng Gang , Shen Lu , Tian Yuejie , Zhou Zhijun , Li Xinlin , Du Xiaoxi
Published: April 2015

Background

The monitoring of cosmetic adverse reactions (CARs) in China has progressed to a new stage of development because of government institutional reforms.

Objective

To discuss and formulate an appropriate model for CAR monitoring in China.

Methods

The formulation of a new CAR‐monitoring model under the administration of the Food and Drug Supervision Department is discussed on the basis of China's current national conditions and with reference to the previous CAR‐monitoring and drug‐monitoring models of the organization formerly known as the Ministry of Health of the People's Republic of China.

Results

The preliminary design of the CAR‐monitoring model is considered to be a long‐term development goal of CAR monitoring. The monitoring model at this stage was developed on the basis of China's national conditions; specifically, the development of a two‐level (national and provincial agencies) monitoring system that relies on CAR‐monitoring sentinels, and reports that cover CARs with definite causal associations.

Conclusion

This model is developed on both theoretical and practical foundations, and can function as a proper monitoring model at the initial stage of current CAR monitoring through an appropriate pilot programme, exploration, and improvement.
Surgery for rectal cancer after high‐dose radiotherapy for prostate cancer: is sphincter preservation relevant?
Authors: Buscail E. , Blondeau V. , Adam J.‐P. , Pontallier A. , Laurent C. , Rullier E. , Denost Q.
Published: April 2015

Aim

The feasibility and outcome of sphincter‐saving resection for rectal cancer were assessed in patients previously treated by high‐dose radiotherapy for prostate cancer.

Method

Between 2000 and 2012, 1066 patients underwent rectal excision for rectal cancer. Of these, 236 were treated by conventional radiotherapy (45 Gy) and sphincter‐saving resection (Group A) and 12 were treated by external‐beam radiotherapy (EBRT) for prostate cancer (70 Gy) and sphincter‐saving resection (Group B) of whom five had a metachronous and seven a synchronous cancer. The end‐points were surgical morbidity, pelvic sepsis, reoperation and definitive stoma.

Results

Tumour characteristics were similar in both groups. Surgical morbidity (67% vs 25%, P = 0.004), anastomotic leakage (50% vs 10%, P = 0.001, and reoperation (50% vs 17%, P = 0.011) were significantly higher in Group B. Multivariate analyses showed that EBRT for prostate cancer was the only independent factor for anastomotic leakage (OR = 5.12; 95% CI 1.45–18.08; P = 0.011) and definitive stoma (OR = 10.56; 95% CI 3.02–39.92; P < 0.001).

Conclusion

High‐dose radiotherapy for prostate cancer increases morbidity from rectal surgery and the risk of a permanent stoma. This suggests that a delayed coloanal anastomosis or a Hartmann procedure should be proposed as an alternative to low anterior resection in this population.
Taking the Bitter with the Sweet: Sugarcane's Return as a Driver of Tropical Deforestation
Authors: Obidzinski Krystof , Kusters Koen , Gnych Sophia
Published: April 2015

Over more than 400 years, large areas of tropical forest in Brazil, the Caribbean, the Philippines, Australia, and other parts of the world were cleared to make way for sugarcane plantations. There is a general consensus in the scientific community that since the 1950s, the frontier expansion of sugarcane has stabilized and direct pressure on tropical forests from sugarcane expansion has diminished. Here, we show, however, that sugarcane plantations are on the cusp of returning as a major driver of deforestation in Indonesia. The Indonesian government has developed preferential policies designed to boost sugar production in the name of national food security, and is seeking to convert more than 1 million hectares of tropical forest into sugarcane plantations. If fully developed, the plantation expansion program will undermine Indonesia's goal of reducing greenhouse gas emissions. The scale of the expansion program is such that it will radically alter the global environmental impact of sugarcane.
Use of IVIg to identify potential miRNA targets for allograft rejection and GvHD therapy
Authors: Padet Lauriane , Loubaki Lionel , Bazin Renée
Published: April 2015

Allograft rejection (AR) and graft‐versus‐host disease (GvHD) are serious complications following transplantation. Micro‐RNAs (miRNAs) have recently been identified as key players in the regulation of these disorders. Because intravenous immunoglobulin (IVIg) has shown therapeutic potential for the prophylaxis and post‐transplant reduction of AR and GvHD, we hypothesized that the effect of IVIg could result from the modulation of specific miRNA expression. To identify such miRNA, we performed mixed lymphocyte reactions (MLRs) as an in vitro model of AR and GvHD, with or without IVIg. We herein show that IVIg strongly inhibits the MLRs. This inhibition is associated with a modulation in the expression of miRNAs implicated in the regulation of pro‐inflammatory cytokine (IL‐2, IL‐6, IFN‐γ) and costimulatory molecule (CD80) expression. We propose that these identified miRNAs could represent potential therapeutic targets for the prevention and therapy of AR and GvHD.