Phe393 mutants of cytochrome P450 BM3 with modified heme redox potentials have altered heme vinyl and propionate conformations.
It has been well established that the heme redox potential is affected by many different factors. Among others, it is sensitive to the proximal heme ligand and the conformation of the propionate and vinyl groups. In the cytochrome P450 BM3 heme domain, substitution of the highly conserved phenylalanine 393 results in a dramatic change in the heme redox potential [Ost, T. W. B., Miles, C. S., Munro, A. W., Murdoch, J., Reid, G. A., and Chapman, S. K. (2001) Biochemistry 40, 13421-13429]. We have used resonance Raman spectroscopy to characterize heme structural changes and modification of heme interactions with the protein matrix that are induced by the F393 substitutions and to determine their correlation with the heme redox potential. Our results show that the Fe-S stretching frequency of the 5-coordinated, high-spin ferric heme is not affected by the mutations, suggesting that the electron density in the Fe-S bond in this state is not affected by the F393 mutation and is not a good indicator of the heme redox potential. Substrate binding perturbs the hydrogen bonding between one propionate group and the protein matrix and correlates to both the size of residue 393 and the heme redox potential. However, heme reduction does not affect the conformation of the propionate groups. Although the conformation of the vinyl groups is not affected much by substrate binding, their conformation changes from mainly out-of-plane to predominantly in-plane upon heme reduction. The extent of these conformational changes correlates strongly with the size of the 393 residue and the heme redox potential, suggesting that steric interaction between this residue and the vinyl groups may be of importance in regulating the heme redox potential in the P450 BM3 heme domain. Further implications of our findings for the change in redox potential upon mutation of F393 will be discussed.


A new specimen of Prolacerta broomi from the lower Fremouw Formation (Early Triassic) of Antarctica, its biogeographical implications and a taxonomic revision.
Prolacerta broomi is an Early Triassic archosauromorph of particular importance to the early evolution of archosaurs. It is well known from many specimens from South Africa and a few relatively small specimens from Antarctica. Here, a new articulated specimen from the Fremouw Formation of Antarctica is described in detail. It represents the largest specimen of Prolacerta described to date with a nearly fully articulated and complete postcranium in addition to four skull elements. The study of this specimen and the re-evaluation of other Prolacerta specimens from both Antarctica and South Africa reveal several important new insights into its morphology, most notably regarding the premaxilla, manus, and pelvic girdle. Although well-preserved skull material from Antarctica is still lacking for Prolacerta, a detailed comparison of Prolacerta specimens from Antarctica and South Africa corroborates previous findings that there are no characters clearly distinguishing the specimens from these different regions and therefore the Antarctic material is assigned to Prolacerta broomi. The biogeographical implications of these new findings are discussed. Finally, some osteological characters for Prolacerta are revised and an updated diagnosis and phylogenetic analysis are provided.


Whose clock makes yours tick? How maternal cardiorespiratory physiology influences newborns' heart rate variability.
This study examined the existence of direct maternal-infant physiological relatedness in respiratory sinus arrhythmia (RSA) when the infant was age 1, 2, 4, 8, and 12 weeks. We instructed mothers to breathe at 6, 12, 15, 20, and 6 cycles per minute while their infants lay on their body. The mother-infant ECG and respiration were registered and video recordings were made. RR-interval (RRI), respiration rate (fR) and RSA were calculated and mother-infant RSA response-patterns were analyzed. The results revealed that infants adjusted their RSA levels to their mothers' levels during the first 2 months of life, but not at 3 months of age, which could be interpreted as a continuing intra-uterine effect. The attenuation between 2 and 3 months could be a reflection of the 2-month developmental shift of social orientation.


Post-Messinian evolutionary relationships across the Sicilian channel: mitochondrial and nuclear markers link a new green toad from Sicily to African relatives.
Little attention has been paid to the consequences of the last landbridge between Africa and Sicily on Mediterranean biogeography. Previous paleontological and scarce molecular data suggest possible faunal exchange later than the well-documented landbridge in the Messinian (5.3 My); however, a possible African origin of recent terrestrial Sicilian fauna has not been thoroughly tested with molecular methods. To gain insight into the phylogeography of the region, we examine two mitochondrial and two nuclear markers (one is a newly adapted intron marker) in green toads (Bufo viridis subgroup) across that sea barrier, the Strait of Sicily. Extensive sampling throughout the western Mediterranean and North Africa revealed a deep sister relationship between Sicilian (Bufo siculus n.sp.) and African green toads (B. boulengeri) on the mitochondrial and nuclear level. Divergence times estimated under a Bayesian-coalescence framework (mtDNA control region and 16S rRNA) range from the Middle Pliocene (3.6 My) to Pleistocene (0.16 My) with an average (1.83 to 2.0 My) around the Pliocene/Pleistocene boundary, suggesting possible land connections younger than the Messinian (5.3 My). We describe green toads from Sicily and some surrounding islands as a new endemic species (Bufo siculus). Bufo balearicus occurs on some western Mediterranean islands (Corsica, Sardinia, Mallorca, and Menorca) and the Apennine Peninsula, and is well differentiated on the mitochondrial and nuclear level from B. siculus as well as from B. viridis (Laurenti), whose haplotype group reaches northeastern Italy, north of the Po River. Detection of Calabrian B. balearicus haplotypes in northeastern Sicily suggests recent invasion. Our data agree with paleogeographic and fossil data, which suggest long Plio-Pleistocene isolation of Sicily and episodic Pleistocene faunal exchange across the Strait of Messina. It remains unknown whether both species (B. balearicus, B. siculus) occur in sympatry in northern Sicily. Our findings on green toads give the first combined mitochondrial and nuclear sequence evidence for a phylogeographic connection across the Strait of Sicily in terrestrial vertebrates. These relationships may have implications for comparative phylogeographic research on other terrestrial animals co-occurring in North Africa and Sicily.


A look at the problem of falls among people with cancer.
People with cancer often have multiple risk factors for falls. A major risk factor is aging; because people often are diagnosed with cancer at an older age or are living longer with cancer, many are at risk for falls. Additional risk factors include impaired physical function, sensory-neurologic deficits, use of multiple medications, and deconditioning, often caused by treatment-induced fatigue. Of all types of accidents, falls pose the most serious threat to the elderly. Fall injuries can have serious consequences related to reduced physical functioning and quality of life. A detailed understanding and awareness of the risk factors (both intrinsic and extrinsic) and problems associated with falls can assist oncology nurses to be vigilant in the identification of patients at high risk for falls. These patients may benefit from protocols used in other patient populations, new interventions, or the use of assistive devices.


Proportion congruency and practice: A contingency learning account of asymmetric list shifting effects.
Performance is impaired when a distracting stimulus is incongruent with the target stimulus (e.g., "green" printed in red). This congruency effect is decreased when the proportion of incongruent trials is increased, termed the proportion congruent effect. This effect is typically interpreted in terms of the adaptation of attention in response to conflict. In contrast, the contingency account argues that the effect is driven by the learning of predictive relationships between words and responses. In a recent report, Abrahamse, Duthoo, Notebaert, and Risko (2013) demonstrated larger changes in the magnitude of the proportion congruent effect when switching from a mostly congruent list to a mostly incongruent list, relative to the reverse order. They argued that this asymmetric list shifting effect fits only with the conflict adaptation perspective. However, the current paper presents reanalyses of this data and an adaptation of the Parallel Episodic Processing model that together demonstrate how the contingency account can explain these findings equally well when considering the generally accepted notion that performance improves with practice. The contingency account may still be the most parsimonious view. (PsycINFO Database Record


Pulsed radiofrequency application in the treatment of chronic pain.
The efficacy of pulsed radiofrequency (PRF) in the treatment of painful lumbosacral spondylosis has been reported. This case series reviews 22 consecutive patients presenting to clinic who had been previously treated with PRF with good results. Patients being prescribed opioids were excluded. During the PRF application, tissue temperature was limited to 43 degrees C. A minimum of 200 mA of current was delivered in each case. The minimum current (at 50 Hz) necessary to stimulate the involved nerve was recorded. The duration of time from PRF treatment until the patient requested a subsequent application was documented. The effective duration of PRF in patients treated for lumbosacral spondylosis ranged from 5 to 18 months (mean +/- SD: 9 +/- 3.7 months; n = 16). PRF applications to dorsal root ganglia were effective from 2 to 12 months (7 +/- 3.8 months; n = 8). Similar results were observed when PRF was applied to cervical medial branch nerves, one suprascapular nerve, and one stellate ganglion. The mean (50 Hz) sensory stimulation thresholds obtained before treatment ranged from 0.08 V to 0.14 V. In this select population of patients not receiving controlled substances, who had a favorable response to a previous PRF application, the duration of pain relief supports the use of PRF as an effective pain treatment.


Long-term survival in metastasized leiomyosarcoma: a case report and review of the literature.
Leiomyosarcoma of the mesosigma is a very rare entity, with low 5-year survival rates. Treatment consists of resection of the primary tumor and, if applicable, of synchronous or metachronous metastases. Local treatment options for metastatic disease should be exploited as long as possible, as response to chemotherapy is reportedly disappointing. Stereotactic radiotherapy is a fairly new locally effective treatment option which has been well established in stereotactic radiotherapy of lung tumors. Whether repeated stereotactic radiotherapy sessions for treatment of lung metastases can be safely and successfully performed over a long time period is not yet well documented. We present the case of a 71-year-old female patient who had a primary diagnosis of lung metastases 12 years ago. Atypical resections of 4 lung metastases were performed in 2001 and 2002. Between 2004 and 2011, 7 sessions of stereotactic body irradiation of lung metastases were performed. All stereotactic treatment were tolerated well (no radiation pneumonitis, FEV1 was 1.3 L [67.8%] in 2004 and 0.99 L [56.3%] in 2011). The present case could demonstrate that a repetitive treatment of lung metastases with multiple stereotactic radiotherapy sessions can lead to long-term survival with a good quality of life.


Drug absorption from large intestine: physicochemical factors governing drug absorption.
Profiles of absorption versus drug molecular weight and absorption versus drug lipophilicity were investigated in both the small and large intestines of rats by an in situ loop method. The absorption-molecular weight profiles examined using different-sized polyethylene glycols (PEGs) were different between the small and large intestines; the large-intestinal absorption of PEGs with molecular weights larger than 300 was poor, while PEGs with molecular weights up to 600 were relatively well absorbed in the small intestine. It is suggested that the paracellular route for drug penetration in the large intestine is restricted more than in the small intestine. The absorption-lipophilicity profiles were also examined in various regions (loops of 6 cm) of rat intestine using three acylsalicylic acids, acetyl-, propionyl- and butyrylsalicylic acids. The absorption rates of the acylsalicylic acids were different in the intestinal regions: the jejunum > the ileum > the colon > the rectum. In each region, the absorption rate increased with the drug lipophilicity. However, it was shown that the absorption rates in the small intestine tended to reach a ceiling at the high lipophilicity. To confirm this tendency, the absorption rates of acetaminophen and indomethacin were compared in the four intestinal regions. The absorption rates of highly lipophilic indomethacin were similar in the large and small intestines, while intermediately lipophilic acetaminophen was more rapidly absorbed in the small intestine than in the large intestine. A thicker unstirred water layer adjacent to the small-intestinal mucosa would be one of the factors which cause such varying absorption-lipophilicity profiles.


An evaluation of ribonuclease protection assays for the detection of beta-cardiac myosin heavy chain gene mutations.
Ribonuclease (RNase) protection has been used to identify beta-cardiac myosin heavy chain (MHC) gene mutations that cause familial hypertrophic cardiomyopathy (FHC). Since more than 10 different mutations within this gene have been demonstrated to cause FHC in unrelated individuals, the genetic diagnosis of this condition will involve screening the beta-MHC gene. The accuracy with which RNase protection identifies such mutations is critical to defining the utility of this methodology in detecting mutations that cause FHC. Twelve unrelated individuals with FHC were selected for further study because their beta-MHC genes had been screened for mutations by use of RNase protection, and no mutation was found. We performed linkage analysis of the families of these 12 probands using polymorphic short tandem repeats within the beta-MHC gene to determine whether FHC was genetically linked to the MHC locus on chromosome 14. FHC was not genetically linked to the MHC locus in 11 families whose beta-cardiac MHC gene did not contain mutations detectable by RNase protection. We conclude that RNase protection is a sensitive method for screening for mutations within the beta-cardiac MHC gene. Further, mutations in the noncoding regions of the beta-MHC gene and mutations in the alpha-cardiac MHC gene are not a common cause of FHC. Negative RNase protection assays of affected individuals suggest that their FHC is due to mutations at other loci.


Resource utilisation for neuropsychiatric disorders in developing countries: a multinational Delphi consensus study.
Substantial information is now available at a global and regional level relating to the burden of mental disorders, yet hardly any international data are available on the resources that are needed to respond to this global burden. The aim of this study was to develop consensus-driven estimates of resource utilisation, coverage and adherence for key mental health care interventions capable of reducing leading neuropsychiatric causes of disease burden. A resource utilisation questionnaire was developed for a series of mental health care interventions for schizophrenia, affective and anxiety disorders, alcohol misuse, epilepsy and Alzheimer's disease. Expert opinion surveys using a Delphi approach were conducted in seven countries (Brazil, China, Egypt, Ghana, India, Mexico and Thailand). Initial results were fed back to participants in order to elicit a second, final set of consensus estimates. Mean and median values were calculated, together with measures of dispersion and agreement. Sixty-seven psychiatrists took part in the first Delphi round and 60 completed the seven consensus panels in the second Delphi round. Expected resource use was highest for acute psychosis, mania, severe depression and severe dementia (at least monthly outpatient visits, and over 25% of cases admitted to hospital with an average length of stay one month). Aggregated estimates for current treatment coverage ranged from below 20% for Alzheimer's disease to 60% for epilepsy. Coverage and adherence levels for affective disorders were both below 50%. The Delphi technique proved to be a feasible and efficient method for data collection and consensus generation concerning mental health care resource utilisation. Results provide a set of range estimates for mental health resource planning and evaluation in a variety of developing regions.


The Nogo-C2/Nogo receptor complex regulates the morphogenesis of zebrafish lateral line primordium through modulating the expression of dkk1b, a Wnt signal inhibitor.
The fish lateral line (LL) is a mechanosensory system closely related to the hearing system of higher vertebrates, and it is composed of several neuromasts located on the surface of the fish. These neuromasts can detect changes in external water flow, to assist fish in maintaining a stationary position in a stream. In the present study, we identified a novel function of Nogo/Nogo receptor signaling in the formation of zebrafish neuromasts. Nogo signaling in zebrafish, like that in mammals, involves three ligands and four receptors, as well as three co-receptors (TROY, p75, and LINGO-1). We first demonstrated that Nogo-C2, NgRH1a, p75, and TROY are able to form a Nogo-C2 complex, and that disintegration of this complex causes defective neuromast formation in zebrafish. Time-lapse recording of the CldnB::lynEGFP transgenic line revealed that functional obstruction of the Nogo-C2 complex causes disordered morphogenesis, and reduces rosette formation in the posterior LL (PLL) primordium during migration. Consistent with these findings, hair-cell progenitors were lost from the PLL primordium in p75, TROY, and Nogo-C2/NgRH1a morphants. Notably, the expression levels of pea3, a downstream marker of Fgf signaling, and dkk1b, a Wnt signaling inhibitor, were both decreased in p75, TROY, and Nogo-C2/NgRH1a morphants; moreover, dkk1b mRNA injection could rescue the defects in neuromast formation resulting from knockdown of p75 or TROY. We thus suggest that a novel Nogo-C2 complex, consisting of Nogo-C2, NgRH1a, p75, and TROY, regulates Fgf signaling and dkk1b expression, thereby ensuring stable organization of the PLL primordium.


Involvement of human basal ganglia in offline feedback control of voluntary movement.
Practice makes perfect, but the neural substrates of trial-to-trial learning in motor tasks remain unclear. There is some evidence that the basal ganglia process feedback-related information to modify learning in essentially cognitive tasks , but the evidence that these key motor structures are involved in offline feedback-related improvement of performance in motor tasks is paradoxically limited. Lesion studies in adult zebra finches suggest that the avian basal ganglia are involved in the transmission or production of an error signal during song . However, patients with Huntington's disease, in which there is prominent basal ganglia dysfunction, are not impaired in error-dependent modulation of future trial performance . By directly recording from the subthalamic nucleus in patients with Parkinson's disease, we demonstrate that this nucleus processes error in trial performance at short latency. Local evoked activity is greatest in response to smallest errors and influences the programming of subsequent movements. Accordingly, motor parameters are least likely to change after the greatest evoked responses so that accurately performed trials tend to precede other accurate trials. This relationship is disrupted by electrical stimulation of the nucleus at high frequency. Thus, the human subthalamic nucleus is involved in feedback-based learning.


A systematic review of studies in the contributions of the work environment to ischaemic heart disease development.
There is need for an updated systematic review of associations between occupational exposures and ischaemic heart disease (IHD), using the GRADE system. (i) publication in English in peer-reviewed journal between 1985 and 2014, (ii) quantified relationship between occupational exposure (psychosocial, organizational, physical and other ergonomic job factors) and IHD outcome, (iii) cohort studies with at least 1000 participants or comparable case-control studies with at least 50 + 50 participants, (iv) assessments of exposure and outcome at baseline as well as at follow-up and (v) gender and age analysis. Relevance and quality were assessed using predefined criteria. Level of evidence was then assessed using the GRADE system. Consistency of findings was examined for a number of confounders. Possible publication bias was discussed. Ninety-six articles of high or medium high scientific quality were finally included. There was moderately strong evidence (grade 3 out of 4) for a relationship between job strain and small decision latitude on one hand and IHD incidence on the other hand. Limited evidence (grade 2) was found for iso-strain, pressing work, effort-reward imbalance, low support, lack of justice, lack of skill discretion, insecure employment, night work, long working week and noise in relation to IHD. No difference between men and women with regard to the effect of adverse job conditions on IHD incidence. There is scientific evidence that employees, both men and women, who report specific occupational exposures, such as low decision latitude, job strain or noise, have an increased incidence of IHD.


Client Perspectives on Desirable Attributes and Skills of Veterinary Technologists in Australia: Considerations for Curriculum Design.
Client or service user perspectives are important when designing curricula for professional programs. In the case of veterinary technology, an emerging profession in the veterinary field in Australasia, client views on desirable graduate attributes, skills, and knowledge have not yet been explored. This study reports on a survey of 441 veterinary clients (with 104 responses) from four veterinary practices in Brisbane, Queensland, conducted between October 2008 and February 2009. The included veterinary practices provided clinical placements for veterinary technology undergraduates and employment for veterinary technology graduates (2003-2007). Client socio-demographic data along with ratings of the importance of a range of technical (veterinary nursing) skills, emotional intelligence, and professional attributes for veterinary technology graduates were collected and analyzed. Overall, the majority of clients viewed technical skills, emotional intelligence, and professional attributes as important in the clinical practice of veterinary technology graduates with whom they interacted in the veterinary practice. Client interviews (n=3) contextualized the survey data and also showed that clients attached importance to graduates demonstrating professional competence. Agglomerative hierarchical cluster analysis revealed four distinct groupings of clients within the data based on their differing perceptions. Using a multivariable proportional-odds regression model, it was also found that some client differences were influenced by demographic factors such as gender, age, and number of visits annually. For example, the odds of female clients valuing emotionality and sociability were greater than males. These findings provide useful data for the design of a professionalizing and market-driven veterinary technology curriculum.


Construction and validation of nano gold tripods for molecular imaging of living subjects.
Anisotropic colloidal hybrid nanoparticles exhibit superior optical and physical properties compared to their counterparts with regular architectures. We herein developed a controlled, stepwise strategy to build novel, anisotropic, branched, gold nanoarchitectures (Au-tripods) with predetermined composition and morphology for bioimaging. The resultant Au-tripods with size less than 20 nm showed great promise as contrast agents for in vivo photoacoustic imaging (PAI). We further identified Au-tripods with two possible configurations as high-absorbance nanomaterials from various gold multipods using a numerical simulation analysis. The PAI signals were linearly correlated with their concentrations after subcutaneous injection. The in vivo biodistribution of Au-tripods favorable for molecular imaging was confirmed using small animal positron emission tomography (PET). Intravenous administration of cyclic Arg-Gly-Asp-d-Phe-Cys (RGDfC) peptide conjugated Au-tripods (RGD-Au-tripods) to U87MG tumor-bearing mice showed PAI contrasts in tumors almost 3-fold higher than for the blocking group. PAI results correlated well with the corresponding PET images. Quantitative biodistribution data revealed that 7.9% ID/g of RGD-Au-tripods had accumulated in the U87MG tumor after 24 h post-injection. A pilot mouse toxicology study confirmed that no evidence of significant acute or systemic toxicity was observed in histopathological examination. Our study suggests that Au-tripods can be reliably synthesized through stringently controlled chemical synthesis and could serve as a new generation of platform with high selectivity and sensitivity for multimodality molecular imaging.


The evolution of ribosomal DNA: divergent paralogues and phylogenetic implications.
Although nuclear ribosomal DNA (rDNA) repeats evolve together through concerted evolution, some genomes contain a considerable diversity of paralogous rDNA. This diversity includes not only multiple functional loci but also putative pseudogenes and recombinants. We examined the occurrence of divergent paralogues and recombinants in Gossypium, Nicotiana, Tripsacum, Winteraceae, and Zea ribosomal internal transcribed spacer (ITS) sequences. Some of the divergent paralogues are probably rDNA pseudogenes, since they have low predicted secondary structure stability, high substitution rates, and many deamination-driven substitutions at methylation sites. Under standard PCR conditions, the low stability paralogues amplified well, while many high-stability paralogues amplified poorly. Under highly denaturing PCR conditions (i.e., with dimethylsulfoxide), both low- and high-stability paralogues amplified well. We also found recombination between divergent paralogues. For phylogenetics, divergent ribosomal paralogues can aid in reconstructing ancestral states and thus serve as good outgroups. Divergent paralogues can also provide companion rDNA phylogenies. However, phylogeneticists must discriminate among families of divergent paralogues and recombinants or suffer from muddled and inaccurate organismal phylogenies.


Exploration of prognostic biomarkers and therapeutic targets in the microenvironment of bladder cancer based on CXC chemokines.
Bladder cancer (BLCA) has a high rate of morbidity and mortality, and is considered as one of the most malignant tumors of the urinary system. Tumor cells interact with surrounding interstitial cells, playing a key role in carcinogenesis and progression, which is partly mediated by chemokines. CXC chemokines exert anti-tumor biological roles in the tumor microenvironment and affect patient prognosis. Nevertheless, their expression and prognostic values patients with BLCA remain unclear. We used online tools, including Oncomine, UALCAN, GEPIA, GEO databases, cBioPortal, GeneMANIA, DAVID 6.8, Metascape, TRUST (version 2.0), LinkedOmics, TCGA, and TIMER2.0 to perform the relevant analysis. The mRNA levels of C-X-C motif chemokine ligand (CXCL)1, CXCL5, CXCL6, CXCL7, CXCL9, CXCL10, CXCL11, CXCL13, CXCL16, and CXCL17 were increased significantly increased, and those of CXCL2, CXCL3, and CXCL12 were decreased significantly in BLCA tissues as assessed using the Oncomine, TCGA, and GEO databases. GEO showed that high levels of CXCL1, CXCL6, CXCL10, CXCL11, and CXCL13 mRNA expression are associated significantly with the poor overall survival (all p < 0.05), and similarly, those of CXCL2 and CXCL12 in the TCGA database (p < 0.05). The predominant signaling pathways involving the differentially expressed CXC chemokines are cell cycle, chemokine, and cytokine-cytokine receptor interaction. Moreover, transcription factors such as Sp1 transcription factor (SP1), nuclear factor kappa B subunit 1 (NFKB1), and RELA proto-oncogene, NF-KB subunit (RELA) were likely play critical roles in regulating CXC chemokine expression. LYN proto-oncogene, src family tyrosine kinase (LYN) and LCK proto-oncogene, src family tyrosine kinase (LCK) were identified as the key targets of these CXC chemokines. MicroRNAs miR200 and miR30 were identified as the main microRNAs that interact with several CXC chemokines through an miRNA-target network. The expression of these chemokines is closely associated with the infiltration of six categories of immune cells. We explored the CXC chemokines superfamily-based biomarkers associated with BLCA prognosis using public databases, and provided possible chemokine targets for patients with BLCA.


Ornithine decarboxylase knockout in Tapesia yallundae abolishes infection plaque formation in vitro but does not reduce virulence toward wheat.
A knockout strain of Tapesia yallundae lacking the single ornithine decarboxylase (ODC) allele has been created by targeted gene replacement. A central region of the ODC gene was isolated by polymerase chain reaction with degenerate oligonucleotides and used to probe a lambda genomic library. The gene was sequenced, and the encoded ODC protein sequence was shown to be similar to those from other fungi. The functionality of the T. yallundae ODC was confirmed by complementation of an Aspergillus nidulans mutant (puA) strain devoid of ODC activity, restoring growth in the absence of exogenous polyamines. Transformation-mediated gene replacement was used to create strains that were auxotrophic for putrescine and lack ODC coding sequences. ODC knockout strains were unable to differentiate infection structures after in vitro induction and showed an abnormal hyphal branching phenotype. Pathogenicity studies on these mutants showed that, surprisingly, they are not reduced in virulence compared with nondisrupted transformants. This suggests that the strains carrying an ODC disruption can obtain sufficient polyamines from the host plant for normal growth and differentiation and, therefore, that fungal ODC may not be a suitable target for fungicides.


Integrative analysis of multiple gene expression profiles with quality-adjusted effect size models.
With the explosion of microarray studies, an enormous amount of data is being produced. Systematic integration of gene expression data from different sources increases statistical power of detecting differentially expressed genes and allows assessment of heterogeneity. The challenge, however, is in designing and implementing efficient analytic methodologies for combination of data generated by different research groups. We extended traditional effect size models to combine information from different microarray datasets by incorporating a quality measure for each gene in each study into the effect size estimation. We illustrated our method by integrating two datasets generated using different Affymetrix oligonucleotide types. Our results indicate that the proposed quality-adjusted weighting strategy for modelling inter-study variation of gene expression profiles not only increases consistency and decreases heterogeneous results between these two datasets, but also identifies many more differentially expressed genes than methods proposed previously. Data integration and synthesis is becoming increasingly important. We live in a high-throughput era where technologies constantly change leaving behind a trail of data with different forms, shapes and sizes. Statistical and computational methodologies are therefore critical for extracting the most out of these related but not identical sources of data.


Telomerase reverse transcriptase promoter mutations in tumors originating from the adrenal gland and extra-adrenal paraganglia.
Hotspot mutations in the promoter of the telomerase reverse transcriptase (TERT) gene have been recently reported in human cancers and proposed as a novel mechanism of telomerase activation. To explore TERT promoter mutations in tumors originating from the adrenal gland and extra-adrenal paraganglia, a set of 253 tumors (38 adrenocortical carcinomas (ACCs), 127 pheochromocytomas (PCCs), 18 extra-adrenal paragangliomas (ea PGLs), 37 head and neck PGLs (HN PGLs), and 33 peripheral neuroblastic tumors) was selected along with 16 human neuroblastoma (NBL) and two ACC cell lines to assess TERT promoter mutations by the Sanger sequencing method. All mutations detected were confirmed by a SNaPshot assay. Additionally, 36 gastrointestinal stromal tumors (GISTs) were added to explore an association between TERT promoter mutations and SDH deficiency. TERT promoter mutations were found in seven out of 289 tumors and in three out of 18 human cell lines; four C228T mutations in 38 ACCs (10.5%), two C228T mutations in 18 ea PGLs (11.1%), one C250T mutation in 36 GISTs (2.8%), and three C228T mutations in 16 human NBL cell lines (18.75%). No mutation was detected in PCCs, HN PGLs, neuroblastic tumors as well as ACC cell lines. TERT promoter mutations preferentially occurred in a SDH-deficient setting (P=0.01) being present in three out of 47 (6.4%) SDH-deficient tumors vs zero out of 171 (0%) SDH-intact tumors. We conclude that TERT promoter mutations occur in ACCs and ea PGLs. In addition, preliminary evidence indicates a potential association with the acquisition of TERT promoter mutations in SDH-deficient tumors.


[Malocclusions in children and adolescents from villages and native communities in the Ucayali Amazon region in Peru].
We conducted a cross-sectional descriptive study to assess the prevalence of malocclusions in children and adolescents aged 2 to 18 years of villages and native communities of the Ucayali jungle of Peru. We assessed the presence of malocclusions using Angle's classification and orthodontic changes. We evaluated 201 individuals, 106 (52.7%) were women, most of them (54.7%) had between 6 and 12 years. The prevalence of malocclusions was 85.6%, the most prevalent according to Angle's classification was class I (59.6%). Orthodontic alterations were present in 67.2% of cases. The most frequent were dental crowding (28.4%), anterior crossbite (17.4%), exaggerated overjet (8.5%), excessive overbite (5.0%) and anterior open bite (5.0%). We found a high prevalence of malocclusion and orthodontic changes in the evaluated native communities, highlighting the need to implement preventive programs to improve the oral health of these neglected populations.


Defining the electroclinical phenotype and outcome of PCDH19-related epilepsy: A multicenter study.
PCDH19-related epilepsy is an epileptic syndrome with infantile onset, characterized by clustered and fever-induced seizures, often associated with intellectual disability (ID) and autistic features. The aim of this study was to analyze a large cohort of patients with PCDH19-related epilepsy and better define the epileptic phenotype, genotype-phenotype correlations, and related outcome-predicting factors. We retrospectively collected genetic, clinical, and electroencephalogram (EEG) data of 61 patients with PCDH19-related epilepsy followed at 15 epilepsy centers. All consecutively performed EEGs were analyzed, totaling 551. We considered as outcome measures the development of ID, autistic spectrum disorder (ASD), and seizure persistence. The analyzed variables were the following: gender, age at onset, age at study, genetic variant, fever sensitivity, seizure type, cluster occurrence, status epilepticus, EEG abnormalities, and cognitive and behavioral disorders. Receiver operating characteristic curve analysis was performed to evaluate the age at which seizures might decrease in frequency. At last follow-up (median = 12 years, range = 1.9-42.1 years), 48 patients (78.7%) had annual seizures/clusters, 13 patients (21.3%) had monthly to weekly seizures, and 12 patients (19.7%) were seizure-free for ≥2 years. Receiver operating characteristic analysis showed a significant decrease of seizure frequency after the age of 10.5 years (sensitivity = 81.0%, specificity = 70.0%). Thirty-six patients (59.0%) had ID and behavioral disturbances. ASD was present in 31 patients. An earlier age at epilepsy onset emerged as the only predictive factor for ID (P = 0.047) and ASD (P = 0.014). Conversely, age at onset was not a predictive factor for seizure outcome (P = 0.124). We found that earlier age at epilepsy onset is related to a significant risk for ID and ASD. Furthermore, long-term follow-up showed that after the age of 10 years, seizures decrease in frequency and cognitive and behavioral disturbances remain the primary clinical problems.


Measures of response in clinical trials of systemic sclerosis: the Combined Response Index for Systemic Sclerosis (CRISS) and Outcome Measures in Pulmonary Arterial Hypertension related to Systemic Sclerosis (EPOSS).
There have been steady efforts to develop a combined response index for systemic sclerosis (CRISS). A parallel and equally successful effort has been made by an Expert Panel on Outcome Measures in PAH related to Systemic Sclerosis (EPOSS) to measure effect in treatment of pulmonary arterial hypertension of systemic sclerosis (PAH-SSc). CRISS conducted a Delphi process combined with expert review to identify 11 candidate domains for inclusion in a core set of outcomes for SSc clinical trials: soluble biomarkers, cardiac, digital ulcers, gastrointestinal, global health, health related quality of life (HRQOL) and function, musculoskeletal, pulmonary, Raynaud's, renal, and skin. Tools within domains were also agreed upon. Concentrating on one aspect of disease, PAH, EPOSS also conducted a Delphi process and judged the following domains as the most appropriate for randomized controlled trials in PAH-SSc: lung vascular/pulmonary arterial pressure, cardiac function, exercise testing; severity of dyspnea, discontinuation of treatment; quality of life/activities of daily living; global state; and survival. Possible useful tools within each domain were also agreed on. Patient derived, physician derived, and objective measures of response will be included and combined with the idea that each reflects different aspects of PAH (EPOSS) and overall disease (CRISS) although this assumption may not prove true and can be separated if statistically and clinically valid to do so. In either case, prospective studies will require measurement of all domains, and tools are required and will be developed to define appropriate combined measures of response. CRISS and EPOSS are being developed through the OMERACT process. Through Delphi process and literature review significant progress has been made for both indices, and prospective data are being collected.


Palmitoylation of a conserved cysteine in the regulator of G protein signaling (RGS) domain modulates the GTPase-activating activity of RGS4 and RGS10.
RGS4 and RGS10 expressed in Sf9 cells are palmitoylated at a conserved Cys residue (Cys(95) in RGS4, Cys(66) in RGS10) in the regulator of G protein signaling (RGS) domain that is also autopalmitoylated when the purified proteins are incubated with palmitoyl-CoA. RGS4 also autopalmitoylates at a previously identified cellular palmitoylation site, either Cys(2) or Cys(12). The C2A/C12A mutation essentially eliminates both autopalmitoylation and cellular [(3)H]palmitate labeling of Cys(95). Membrane-bound RGS4 is palmitoylated both at Cys(95) and Cys(2/12), but cytosolic RGS4 is not palmitoylated. RGS4 and RGS10 are GTPase-activating proteins (GAPs) for the G(i) and G(q) families of G proteins. Palmitoylation of Cys(95) on RGS4 or Cys(66) on RGS10 inhibits GAP activity 80-100% toward either Galpha(i) or Galpha(z) in a single-turnover, solution-based assay. In contrast, when GAP activity was assayed as acceleration of steady-state GTPase in receptor-G protein proteoliposomes, palmitoylation of RGS10 potentiated GAP activity >/=20-fold. Palmitoylation near the N terminus of C95V RGS4 did not alter GAP activity toward soluble Galpha(z) and increased G(z) GAP activity about 2-fold in the vesicle-based assay. Dual palmitoylation of wild-type RGS4 remained inhibitory. RGS protein palmitoylation is thus multi-site, complex in its control, and either inhibitory or stimulatory depending on the RGS protein and its sites of palmitoylation.


Anncolvar: Approximation of Complex Collective Variables by Artificial Neural Networks for Analysis and Biasing of Molecular Simulations.
The state of a molecular system can be described in terms of collective variables. These low-dimensional descriptors of molecular structure can be used to monitor the state of the simulation, to calculate free energy profiles or to accelerate rare events by a bias potential or a bias force. Frequent calculation of some complex collective variables may slow down the simulation or analysis of trajectories. Moreover, many collective variables cannot be explicitly calculated for newly sampled structures. In order to address this problem, we developed a new package called anncolvar. This package makes it possible to build and train an artificial neural network model that approximates a collective variable. It can be used to generate an input for the open-source enhanced sampling simulation PLUMED package, so the collective variable can be monitored and biased by methods available in this program. The computational efficiency and the accuracy of anncolvar are demonstrated on selected molecular systems (cyclooctane derivative, Trp-cage miniprotein) and selected collective variables (Isomap, molecular surface area).


Role of high-fidelity Escherichia coli DNA polymerase I in replication bypass of a deoxyadenosine DNA-peptide cross-link.
Reaction of bifunctional electrophiles with DNA in the presence of peptides can result in DNA-peptide cross-links. In particular, the linkage can be formed in the major groove of DNA via the exocyclic amino group of adenine (N⁶-dA). We previously demonstrated that an A family human polymerase, Pol ν, can efficiently and accurately synthesize DNA past N⁶-dA-linked peptides. Based on these results, we hypothesized that another member of that family, Escherichia coli polymerase I (Pol I), may also be able to bypass these large major groove DNA lesions. To test this, oligodeoxynucleotides containing a site-specific N⁶-dA dodecylpeptide cross-link were created and utilized for in vitro DNA replication assays using E. coli DNA polymerases. The results showed that Pol I and Pol II could efficiently and accurately bypass this adduct, while Pol III replicase, Pol IV, and Pol V were strongly inhibited. In addition, cellular studies were conducted using E. coli strains that were either wild type or deficient in all three DNA damage-inducible polymerases, i.e., Pol II, Pol IV, and Pol V. When single-stranded DNA vectors containing a site-specific N⁶-dA dodecylpeptide cross-link were replicated in these strains, the efficiencies of replication were comparable, and in both strains, intracellular bypass of the lesion occurred in an error-free manner. Collectively, these findings demonstrate that despite its constrained active site, Pol I can catalyze DNA synthesis past N⁶-dA-linked peptide cross-links and is likely to play an essential role in cellular bypass of large major groove DNA lesions.


Giant sensitivity of long period gratings in transition mode near the dispersion turning point: an integrated design approach.
We report an original design approach based on the modal dispersion curves for the development of long period gratings in transition mode near the dispersion turning point exhibiting ultrahigh refractive index sensitivity. The theoretical model predicting a giant sensitivity of 9900 nm per refractive index unit in a watery environment was experimentally validated with a result of approximately 9100 nm per refractive index unit around an ambient index of 1.3469. This result places thin film coated LPGs as an alternative to other fiber-based technologies for high-performance chemical and biological sensing applications.


Comparative genomic analysis of the microbiome [corrected] of herbivorous insects reveals eco-environmental adaptations: biotechnology applications.
Metagenome analysis of the gut symbionts of three different insects was conducted as a means of comparing taxonomic and metabolic diversity of gut microbiomes to diet and life history of the insect hosts. A second goal was the discovery of novel biocatalysts for biorefinery applications. Grasshopper and cutworm gut symbionts were sequenced and compared with the previously identified metagenome of termite gut microbiota. These insect hosts represent three different insect orders and specialize on different food types. The comparative analysis revealed dramatic differences among the three insect species in the abundance and taxonomic composition of the symbiont populations present in the gut. The composition and abundance of symbionts was correlated with their previously identified capacity to degrade and utilize the different types of food consumed by their hosts. The metabolic reconstruction revealed that the gut metabolome of cutworms and grasshoppers was more enriched for genes involved in carbohydrate metabolism and transport than wood-feeding termite, whereas the termite gut metabolome was enriched for glycosyl hydrolase (GH) enzymes relevant to lignocellulosic biomass degradation. Moreover, termite gut metabolome was more enriched with nitrogen fixation genes than those of grasshopper and cutworm gut, presumably due to the termite's adaptation to the high fiber and less nutritious food types. In order to evaluate and exploit the insect symbionts for biotechnology applications, we cloned and further characterized four biomass-degrading enzymes including one endoglucanase and one xylanase from both the grasshopper and cutworm gut symbionts. The results indicated that the grasshopper symbiont enzymes were generally more efficient in biomass degradation than the homologous enzymes from cutworm symbionts. Together, these results demonstrated a correlation between the composition and putative metabolic functionality of the gut microbiome and host diet, and suggested that this relationship could be exploited for the discovery of symbionts and biocatalysts useful for biorefinery applications.


Influence of paternal and maternal ethnicity and ethnic enclaves on newborn weight.
The association between maternal ethnicity and newborn weight is understood. Less is known about the additional influence of paternal ethnicity and neighbourhood ethnic composition. We studied 692 301 singleton live births of parents of Canadian, Bangladeshi, Sri Lankan, Pakistani, Indian, Filipino, Vietnamese, Korean, Hong Kong or Chinese birthplace. We used multivariable regression to calculate mean (95% CI) birthweight differences between infants of two Canadian-origin parents and (1) foreign-born mother and Canadian-born father, (2) Canadian-born mother and foreign-born father or (3) two foreign-born parents from the same country. We also stratified by high versus low same-ethnic concentration of the parent's residence. We adjusted for gestational age at birth, maternal age, parity, marital status and income quintile. Compared with male and female infants of two Canadian-born parents, those of same-country foreign-born parents weighed 6.2% (-218 g, 95% CI -214 g to -223 g) and 5.6% (-192 g, 95% CI -187 g to -196 g) less, respectively. The largest mean weight difference was among male (8.4% (-297 g, 95% CI -276 g to -319 g)) and female (8.2% (-279 g, 95% CI -262 g to -296 g)) infants of two Bangladeshi parents. Infants of a foreign-born mother and Canadian-born father had weights closest to those of two Canadian-born parents. Residing in an area of high (vs low) same-ethnic concentration was associated with lower birthweight among infants of mixed union couples, but not among those of parents originating from the same country. Paternal and maternal ethnic origin influence newborn weight, which is modified by settlement in a high same-ethnic concentration area only among parents of mixed union.


Occurrence and clinical implications of heterotopic ossification after cervical disc arthroplasty with the Prestige LP Cervical Disc at 2 contiguous levels.
The authors sought to assess the impact of heterotopic ossification (HO) on clinical outcomes and angular range of motion (ROM) after cervical disc arthroplasty (CDA) performed with the Prestige LP Cervical Disc (Prestige LP disc) at 2 levels. HO was assessed and graded from 0 to IV for increasing severity on lateral neutral radiographs at each visit in 209 patients who underwent implantation of Prestige LP discs at 2 cervical levels in a clinical trial with extended 10-year follow-up. ROM was compared by using HO grade, and clinical outcomes were compared between HO subgroups (grade 0-II vs III/IV) based on HO severity at 2 and 10 years after surgery. The grade III/IV HO incidence at either or both index levels was 24.2% (48/198) at 2 years and 39.0% (57/146) at 10 years. No statistical difference was found in overall success; neurological success; or Neck Disability Index (NDI), neck pain, arm pain, or SF-36 Physical Component Summary (PCS) scores between the HO subgroups (grade 0-II vs III/IV) at either 2 or 10 years. The cumulative rate of possible implant-related adverse events (AEs) was higher in patients having grade III/IV HO at 2 years (56.3%) and 10 years (47.8%) compared with those having grade 0-II HO at 2 years (24.4%) and 10 years (17.9%), specifically in 2 subcategories: spinal events and HOs reported by the investigators. No statistical difference was found between the HO subgroups in possible implant-related serious AEs or secondary surgeries at the index or adjacent levels. The average angular ROMs at index levels were lower in subjects with higher-grade HO at 2 and 10 years. The average ROMs at the superior level were 8.8°, 6.6°, 3.2°, and 0.3°, respectively, for the HO grade 0/I, II, III, and IV groups at 10 years, and 7.9°, 6.2°, 3.7°, and 0.6°, respectively, at the inferior level. Radiographically severe (grade III or IV) HO after CDA with the Prestige LP disc at 2 levels did not significantly affect efficacy or safety outcomes (severe AEs or secondary surgeries). However, severe HO, particularly grade IV HO, significantly limited ROM, as expected.


Fine-mapping reveals novel alternative splicing of the dopamine transporter.
The dopamine transporter gene (SLC6A3, DAT) has been implicated in the pathogenesis of numerous psychiatric and neurodevelopmental disorders, including schizophrenia (SZ). We previously detected association between SZ and intronic SLC6A3 variants that replicated in two independent Caucasian samples, but had no obvious function. In follow-up analyses, we sequenced the coding and intronic regions of SLC6A3 to identify complete linkage disequilibrium patterns of common variations. We genotyped 78 polymorphisms, narrowing the potentially causal region to two correlated clusters of associated SNPs localized predominantly to introns 3 and 4. Our computational analysis of these intronic regions predicted a novel cassette exon within intron 3, designated E3b, which is conserved among primates. We confirmed alternative splicing of E3b in post-mortem human substantia nigra (SN). As E3b introduces multiple in-frame stop codons, the SLC6A3 open reading frame is truncated and the spliced product may undergo nonsense mediated decay. Thus, factors that increase E3b splicing could reduce the amount of unspliced product available for translation. Observations consistent with this prediction were made using cellular assays and in post-mortem human SN. In mini-gene constructs, the extent of splicing is also influenced by at least two common haplotypes, so the alternative splicing was evaluated in relation to SZ risk. Meta-analyses across genome-wide association studies did not support the initial associations and further post-mortem studies did not suggest case-control differences in splicing. These studies do not provide a compelling link to schizophrenia. However, the impact of the alternative splicing on other neuropsychiatric disorders should be investigated. © 2010 Wiley-Liss, Inc.


Severely Impaired Cardiorespiratory Fitness in Patients With Recently Decompensated Systolic Heart Failure.
Hospital admission for decompensated heart failure marks a critical inflection point in a patient's health. Despite the improvement in signs or symptoms during hospitalization, patients have a high likelihood of readmission, reflecting a lack of resolution of the underlying condition. Surprisingly, no studies have characterized the cardiorespiratory fitness of such patients. Fifty-two patients (38 [73%] male, age 57 [52 to 65] years, left ventricular ejection fraction 31% [24 to 38]) underwent cardiopulmonary exercise testing 4 (1 to 10) days after hospital discharge, when stable and without overt signs of volume overload. Transthoracic Doppler echocardiography, measurement of N-terminal pro-B-natriuretic peptide, and quality of life were also assessed. Aerobic exercise capacity was severely reduced: peak oxygen consumption (pVO2) was 14.1 (11.2 to 16.3) ml/kg/min. Ventilatory inefficiency as indicated by the minute ventilation carbon dioxide production relation (VE/VCO2 slope) >30 and oxygen uptake efficiency slope <2.0 was noted in 41 (77%) and 39 (75%) patients, respectively. Forty-five (87%) patients had 1 of 2 high-risk features (pVO2 < 14 ml/kg/min or VE/VCO2 >30). Perceived functional capacity, measured by the Duke Activity Status Index, was also severely reduced and correlated with pVO2. N-terminal pro-B-natriuretic peptide levels and early transmitral velocity/early mitral annulus velocity (E/e') ratio at echocardiography showed a modest correlation with lower pVO2. In conclusion, patients with recently decompensated systolic heart failure demonstrate severe impairment in cardiorespiratory fitness, severely limiting quality of life.


Use of movable high-field-strength intraoperative magnetic resonance imaging with awake craniotomies for resection of gliomas: preliminary experience.
Awake craniotomy with electrocortical mapping and intraoperative magnetic resonance imaging (iMRI) are established techniques for maximizing tumor resection and preserving function, but there has been little experience combining these methodologies. To report our experience of combining awake craniotomy and iMRI with a 1.5-T movable iMRI for resection of gliomas in close proximity to eloquent cortex. Twelve patients (9 male and 3 female patients; age, 32-60 years; mean, 41 years) undergoing awake craniotomy and iMRI for glioma resections were identified from a prospective database. Assessments were made of how these 2 modalities were integrated and what impact this strategy had on safety, surgical decision making, workflow, operative time, extent of tumor resection, and outcome. Twelve craniotomies were safely performed in an operating room equipped with a movable 1.5-T iMRI. The extent of resection was limited because of proximity to eloquent areas in 5 cases: language areas in 3 patients and motor areas in 2 patients. Additional tumor was identified and resected after iMRI in 6 patients. Average operating room time was 7.9 hours (range, 5.9-9.7 hours). Compared with preoperative neurological function, immediate postoperative function was stable/improved in 7 and worse in 5; after 30 days, it was stable/improved in 11 and worse in 1. Awake craniotomy and iMRI with a movable high-field-strength device can be performed safely to maximize resection of tumors near eloquent language areas.


Development and evolution of caste dimorphism in honeybees - a modeling approach.
The difference in phenotypes of queens and workers is a hallmark of the highly eusocial insects. The caste dimorphism is often described as a switch-controlled polyphenism, in which environmental conditions decide an individual's caste. Using theoretical modeling and empirical data from honeybees, we show that there is no discrete larval developmental switch. Instead, a combination of larval developmental plasticity and nurse worker feeding behavior make up a colony-level social and physiological system that regulates development and produces the caste dimorphism. Discrete queen and worker phenotypes are the result of discrete feeding regimes imposed by nurses, whereas a range of experimental feeding regimes produces a continuous range of phenotypes. Worker ovariole numbers are reduced through feeding-regime-mediated reduction in juvenile hormone titers, involving reduced sugar in the larval food. Based on the mechanisms identified in our analysis, we propose a scenario of the evolutionary history of honeybee development and feeding regimes.


Two tandemly organized human genes encoding the T-cell gamma constant-region sequences show multiple rearrangement in different T-cell types.
The recent detailed analysis of genes that undergo rearrangement in T cells has shown that the T-cell receptor genes encoding alpha- and beta-chains are involved in specific alterations in T-cell DNA analogous to the immunoglobulin genes. A third type of gene, designated gamma, has been isolated from mouse cytotoxic T lymphocytes, and evidence suggest that the mouse displays very limited diversity in this gene system, having only three variable-region (V) genes and three constant-region (C) genes. The function of the so-called T-cell gamma gene is unknown. We have isolated genomic genes encoding the human homologue of the mouse T-cell gamma gene; as there is no evidence that this T-cell rearranging gene is anything to do with the T3 molecule, we have designated the human T-cell rearranging gene as TRG gamma (ref. 13), to avoid confusion with the T3 gamma-chain, and have shown that the gene locus maps to chromosome 7 in humans. We now report that human DNA contains two tandemly arranged TRG gamma constant-region genes about 16 kilobases apart. These two genes show multiple rearrangement patterns in a variety of T cells, including helper and cytotoxic/suppressor type, as well as in all forms of T-cell leukaemia. Our results indicate variability of this T-cell gene system in man compared with the analogous system in mouse.


Gender Differences in the Authorship of Original Research in Pediatric Journals, 2001-2016.
To examine the gender of authors of original research in 3 high-impact pediatric journals between 2001 and 2016, given the importance of publishing on academic promotion, and to compare authorship gender with the percentage of women on editorial boards and with academic faculty composition. We assessed the prevalence of female first and senior (last-listed) authorship of original research articles published in 3 pediatric-focused journals Pediatrics, JAMA Pediatrics (entitled Archives of Pediatric and Adolescent Medicine until 2013), and The Journal of Pediatrics. We also examined the gender breakdown of the main editors and the broader editorial boards of these journals. In addition, we examined whether junior female faculty co-authored with male or female senior faculty. Of 3895 original articles, 22 were excluded because the gender of either the first or senior author could not be determined from the name. An analysis of authorship by year showed increasing female representation across the selected journals in both first (39.8% in 2001, 57.7% in 2016) and senior (28.6% in 2001, 38.1% in 2016) authors, respectively. Editorial boards also showed increasing female representation (17.8% in 2001 to 39.8% in 2016). Junior female faculty were more likely to co-author with senior female women (female first and last author); the gap remained unchanged despite the increasing number of women entering pediatrics. Women are underrepresented as authors and editors, although the gap is closing. Junior women are less likely to co-author with senior men, which may be a disservice given current gender disparities in promotion and leadership.


Evidence for novel and specialized mycorrhizal parasitism: the orchid Gastrodia confusa gains carbon from saprotrophic Mycena.
We investigated the physiological ecology of the Asian non-photosynthetic orchid Gastrodia confusa. We revealed its mycorrhizal partners by using molecular identification and identified its ultimate nutritional source by analysing carbon and nitrogen natural stable isotope abundances. Molecular identification using internal transcribed spacer and large subunit nrDNA sequences showed that G. confusa associates with several species of litter- and wood-decomposer Mycena fungi. The carbon and nitrogen isotope signatures of G. confusa were analysed together with photosynthetic plant reference samples and samples of the ectomycorrhizal epiparasite Monotropa uniflora. We found that G. confusa was highly enriched in (13)C but not greatly in (15)N, while M. uniflora was highly enriched in both (13)C and (15)N. The (13)C and (15)N signatures of G. confusa were the closest to those of the fruit bodies of saprotrophic fungi. Our results demonstrate for the first time using molecular and mass-spectrometric approaches that myco-heterotrophic plants gain carbon through parasitism of wood or litter decaying fungi. Furthermore, we demonstrate that, several otherwise free-living non-mycorrhizal, Mycena can be mycorrhizal partners of orchids.


Articular cartilage: from formation to tissue engineering.
Hyaline cartilage is the nonlinear, inhomogeneous, anisotropic, poro-viscoelastic connective tissue that serves as friction-reducing and load-bearing cushion in synovial joints and is vital for mammalian skeletal movements. Due to its avascular nature, low cell density, low proliferative activity and the tendency of chondrocytes to de-differentiate, cartilage cannot regenerate after injury, wear and tear, or degeneration through common diseases such as osteoarthritis. Therefore severe damage usually requires surgical intervention. Current clinical strategies to generate new tissue include debridement, microfracture, autologous chondrocyte transplantation, and mosaicplasty. While articular cartilage was predicted to be one of the first tissues to be successfully engineered, it proved to be challenging to reproduce the complex architecture and biomechanical properties of the native tissue. Despite significant research efforts, only a limited number of studies have evolved up to the clinical trial stage. This review article summarizes the current state of cartilage tissue engineering in the context of relevant biological aspects, such as the formation and growth of hyaline cartilage, its composition, structure and biomechanical properties. Special attention is given to materials development, scaffold designs, fabrication methods, and template-cell interactions, which are of great importance to the structure and functionality of the engineered tissue.


Hybrid inorganic-organic capsules for efficient intracellular delivery of novel siRNAs against influenza A (H1N1) virus infection.
The implementation of RNAi technology into the clinical practice has been significantly postponing due to the issues regarding to the delivery of naked siRNA predominantly to target cells. Here we report the approach to enhance the efficiency of siRNA delivery by encapsulating the siRNA into new carrier systems which are obtained via the combination of widely used layer-by-layer technique and in situ modification by sol-gel chemistry. We used three types of siRNAs (NP-717, NP-1155 and NP-1496) in encapsulated form as new therapeutic agents against H1N1 influenza virus infection. By employing the hybrid microcontainers for the siRNA encapsulation we demonstrate the reduction of viral nucleoprotein (NP) level and inhibition of influenza virus production in infected cell lines (MDCK and A549). The obtained hybrid carriers based on assembled biodegradable polyelectrolytes and sol-gel coating possess several advantages such as a high cell uptake efficiency, low toxicity, efficient intracellular delivery of siRNAs and the protection of siRNAs from premature degradation before reaching the target cells. These findings underpin a great potential of versatile microencapsulation technology for the development of anti-viral RNAi delivery systems against influenza virus infection.


Pulling geometry-induced errors in single molecule force spectroscopy measurements.
In AFM-based single molecule force spectroscopy, it is tacitly assumed that the pulling direction coincides with the end-to-end vector of the molecule fragment being stretched. By systematically varying the position of the attachment point on the substrate relative to the AFM tip, we investigate empirically and theoretically the effect of the pulling geometry on force-extension characteristics of double-stranded DNA. We find that increasing the pulling angle can significantly lower the force of the characteristic overstretching transition and increase the width of the plateau feature beyond the canonical 70%. These effects, when neglected, can adversely affect the interpretation of measured force-extension relationships. We quantitatively evaluate force and extension errors originating from this "pulling angle effect" and stress the need to correct the pulling geometry when stretching rigid molecules with an AFM.


Validation of immune-complex enzyme immunoassays for diagnosis of pneumococcal pneumonia among adults in Kenya.
The efficacy of pneumococcal vaccines in protecting against pneumococcal pneumonia can feasibly be measured only with a diagnostic technique that has a high specificity (0.98 to 1.00) and a sensitivity greatly exceeding that of blood cultures (>0.2 to 0.3). In this context immune-complex enzyme immunoassays (EIAs) offer a novel, convenient diagnostic method, and we have investigated three such assays with appropriate study populations in Kenya. Sera from 129 Kenyan adults with pneumococcal pneumonia and 97 ill controls from the same clinics, but without pneumococcal disease syndromes, were assayed with immune-complex EIAs for pneumolysin, C-polysaccharide, and mixed capsular polysaccharides (Pneumovax II). At an optical density (OD) threshold yielding a specificity of 0.95, the sensitivities (95% confidence intervals) of the assays were 0.22 (0.15 to 0.30), 0.26 (0.19 to 0.34), and 0.22 (0.15 to 0.29), respectively. For pneumolysin immune complexes, human immunodeficiency virus (HIV)-positive patients had a higher mean OD than HIV-negative patients (639 versus 321; P < 0.0001), but stratification by HIV infection status did not alter the performance of this test. Combining the results of all three EIAs did not enhance the diagnostic performances of the individual assays. In Kenyan adults the sensitivities of the immune-complex EIAs could exceed that of blood cultures only at levels of specificity that were insufficient for the performance of vaccine efficacy studies.


Glucagon metabolism in normal subjects and in cirrhotic patients before and after portasystemic venous shunt surgery.
The effect of portasystemic shunt surgery on basal immunoreactive glucagon (IRG) levels, metabolic clearance rate (MCR) and t 1/2 for glucagon decay, and basal systemic delivery rate (BSDR) of glucagon was investigated in paired studies in ten cirrhotic subjects. The degree of hepatocellular dysfunction and extent of portasystemic venous shunting was also recorded. Basal IRG levels were highest in the post-shunt (mean +/- SEM, 382 +/- 73 pg/ml) as compared to the pre-shunt (213 +/- 27 pg/ml; P less than 0.05) cirrhotic and control (53 +/- 13 pg/ml; P less than 0.005) groups. The MCR of glucagon was similar in control (13.0 +/- 1.3 ml/kg/min) and pre-shunt cirrhotic patients (13.3 +/- 1.7 ml/kg/min) but was significantly (P less than 0.02) decreased in the post-shunt cirrhotics (7.6 +/- 1.3 ml/kg/min). The t 1/2 for glucagon decay was similar in the control and cirrhotic groups. The BSDR, an estimate of pancreatic A cell secretion, was increased four-fold (P less than 0.01) in the pre-shunt (3042 +/- 454 pg/kg/min) and post-shunt (2518 +/- 535 pg/kg/min) cirrhotic groups, as compared to controls (750 +/- 244 pg/kg/min). It is concluded that (a) in the presence of cirrhosis, the magnitude of portasystemic shunting is important in determining the degree of hyperglucagonaemia; (b) in preshunt cirrhotics raised basal IRG levels are principally due to A cell hypersecretion of glucagon whereas in post-shunt cirrhotics riased IRG levels reflect both A cell hypersecretion and delayed clearance of glucagon; and (c) acute shunting of splanchnic venous blood away from the liver reduces the clearance of glucagon, suggesting that, in man, the liver contributes to the clearance of circulating glucagon.


Robust conductance of dumbbell molecular junctions with fullerene anchoring groups.
The conductance of a molecular wire connected to metallic electrodes is known to be sensitive to the atomic structure of the molecule-metal contact. This contact is to a large extent determined by the anchoring group linking the molecular wire to the metal. It has been found experimentally that a dumbbell construction with C(60) molecules acting as anchors yields more well-defined conductances as compared to the widely used thiol anchoring groups. Here, we use density functional theory to investigate the electronic properties of this dumbbell construction. The conductance is found to be stable against variations in the detailed bonding geometry and in good agreement with the experimental value of G=3×10(-4) G(0). Electron tunneling across the molecular bridge occurs via the lowest unoccupied orbitals of C(60) which are pinned close to the Fermi energy due to partial charge transfer. Our findings support the original motivation to achieve conductance values more stable towards changes in the structure of the molecule-metal contact leading to larger reproducibility in experiments.


[Potential distribution of jaguar, Panthera onca (Carnivora: Felidae) in Guerrero, Mexico: persistence of areas for its conservation].
Studies about the permanence of natural protected areas are important, because they contribute to the promotion of the conservation target and to optimize economical and human resources of specific areas. Although there are no natural protected areas in Guerrero, it has suitable habitat for the jaguar, a common species used for planning and management of conservation areas. Since, there is actual evidence that environmental and anthropogenic variables may modify vertebrate species distribution with time, in this study we predicted the potential distribution of Panthera onca using MaxEnt for this Southeastern region. In addition, we made a projection considering the effect of a moderate climate change scenario, to evaluate the stability of the conservation area for a period of 24 years. Furthermore, we applied three threat scenarios for the actual prediction to define conservation priorities areas. In our results, we have found that 18 361Km2 (29%) of this state has a permanent suitable habitat for jaguar conservation in the Sierra Madre del Sur and Pacific coast, with a possible loss of 2 000km2 in 24 years. This habitat is characterized by a 56% of temperate forest (mainly conifers and hardwoods 34%), and 35% of tropical deciduous forest. With the projections, the Southeastern region resulted with the higher anthropogenic impacts, while at the same time, an area of 7 900km2 in the Central-Western state was determined as a priority for conservation. To assure jaguar conservation, we propose the inclusion of this new conservation area, which is located in the Sierra Madre del Sur, with which we may potentially preserve other 250 species of threatened vertebrates. This way, the suggested habitat conservation may represent a local effort in Guerrero and will strengthen the biological corridor network for P. onca protection in Latin America.


Transcription factor profiling shows new ways towards new treatment options of cutaneous T cell lymphomas.
Most oncogenes encode activators of transcription factors or transcription factors themselves. Transcription factors that are induced by growth stimuli are, in contrast to transcription factors that regulate house keeping genes, tightly regulated and only active, when a stimulus (e.g. cytokines or other growth factors) is given. Examples of such transcription factors are members of the jun, fos, myc, NFkB and STAT gene families. In cancer cells this regulation is interrupted, resulting in constitutive activities of transcription factors that are normally silent. This in turn results in the increased expression of target genes that are necessary for growth and protection from apoptosis. Since inducible transcription factors are activated by specific pathways, the identification of unusual constitutively active transcription factors also identifies the involved signal transduction pathway. Inhibitors of the components of these pathways may be effective anti-cancer agents, as they interrupt the abnormal signalling and in cancer cells. We applied this strategy for two forms of cutaneous T cell lymphomas and identified several groups of agents that may be the prototypes of new drugs to fight these diseases.


Activation of extracellular signal-regulated kinase signaling by epidermal growth factor mediates c-Jun activation and p300 recruitment in keratin 16 gene expression.
In studies of gene regulation of keratin 16, we reported previously that simian virus 40 promoter factor 1 shows a functional cooperation with c-Jun and coactivators p300/CBP in driving the transcriptional regulation of epidermal growth factor (EGF)-induced keratin 16 gene expression. In the present study, we found that the stimulated expression of keratin 16 by EGF was mediated mainly through the mitogen-activated protein kinase kinase-extracellular signal-regulated kinases 1 and 2 (ERK1/2) signaling pathway. Ser63 and Ser73 on the c-Jun NH(2)-terminal transactivation domain could be phosphorylated in cells treated with EGF; nevertheless, we found that the c-Jun COOH terminus played a pivotal role in EGF-induced expression of keratin 16. The activation of keratin 16 by EGF treatment could not be enhanced by the overexpression of myc-c-JunK3R, in which three putative acetylation lysine residues on the c-Jun COOH terminus were all mutated into arginines, suggesting that c-Jun acetylation on the COOH terminus might partially play a functional role in this system. In addition, by using a chromatin immunoprecipitation assay and a DNA affinity precipitation assay, EGF treatment up-regulated the p300 recruitment through ERK signaling to the promoter region in regulating keratin 16 transcriptional activity. Furthermore, the enhancement of acetyl-histone H3 to the keratin 16 chromatin promoter induced by EGF was also mediated via ERK activation. In conclusion, these results strongly suggest that both c-Jun induction and p300 recruitment to gene promoter, mediated through ERK activation, played an essential role in regulating keratin 16 gene expression by EGF. p300 mediated and regulated EGF-induced keratin 16 gene expression, at least in part, through multiple mechanisms, including a selective acetylation of c-Jun and histone H3.


Calling, courtship, and condition in the fall field cricket, Gryllus pennsylvanicus.
Theoretically, sexual signals should provide honest information about mating benefits and many sexually reproducing species use honest signals when signalling to potential mates. Male crickets produce two types of acoustic mating signals: a long-distance mate attraction call and a short-range courtship call. We tested whether wild-caught fall field cricket (Gryllus pennsylvanicus) males in high condition (high residual mass or large body size) produce higher effort calls (in support of the honest signalling hypothesis). We also tested an alternative hypothesis, whether low condition males produce higher effort calls (in support of the terminal investment hypothesis). Several components of long-distance mate attraction calls honestly reflected male body size, with larger males producing louder mate attraction calls at lower carrier frequencies. Long-distance mate attraction chirp rate dishonestly signalled body size, with small males producing faster chirp rates. Short-range courtship calls dishonestly reflected male residual mass, as chirp rate and pulse rate were best explained by a curvilinear function of residual mass. By producing long-distance mate attraction calls and courtship calls with similar or higher effort compared to high condition males, low condition males (low residual mass or small body size) may increase their effort in current reproductive success at the expense of their future reproductive success, suggesting that not all sexual signals are honest.


Heterogeneous Interactions Promote Crystallization and Spontaneous Resolution of Chiral Molecules.
Predicting the crystallization of chiral molecules from solution is a major challenge in the chemical sciences. In this paper, we use molecular dynamics computer simulations to study the crystallization of a family of coarse-grained models of chiral molecules with a broad range of molecular shapes and interactions. Our simulations reproduce the experimental crystallization behavior of real chiral molecules, including racemic and enantiopure crystals, as well as amorphous solids. Using efficient algorithms for the packing of shapes, we enumerate millions of low-energy crystal structures for each model and analyze the thermodynamic landscape of polymorphs. In agreement with recent conjectures, our analysis shows that the ease of crystallization is largely determined by the number of competing polymorphs with low free energy. We find that this number and, hence, crystallization outcomes depend on molecular interactions in a simple way: Strongly heterogeneous interactions across molecules promote crystallization and favor the spontaneous resolution of racemic mixtures. Our results help rationalize a number of experimental observations and can provide guidance for the design of molecules and experimental conditions for desired crystallization outcomes.


Burden of Infections Among Chronic Myeloid Leukemia Patients Receiving Dasatinib or Nilotinib: A Real-World Retrospective Healthcare Claims Study in the United States.
Tyrosine kinase inhibitors (TKI) have been demonstrated to prolong survival in patients with chronic myeloid leukemia (CML). However, TKIs may be associated with an increased risk of infections. This study compared healthcare resource utilization (HRU) and costs among patients with CML receiving dasatinib or nilotinib, with a focus on infection-related economic outcomes. Two large administrative databases were used to identify adult patients newly diagnosed with CML who initiated dasatinib or nilotinib as first- (1L) or second-line (2L) therapy and were classified into the following 1L (dasatinib 1L/nilotinib 1L cohorts) or 2L (dasatinib 2L/nilotinib 2L) cohorts based on the initiated 1L/2L TKI therapy. Infection-related HRU and healthcare costs were compared between cohorts, separately for 1L and 2L. Cohorts included 1156 patients in the dasatinib 1L and 677 patients in the nilotinib 1L cohorts, 322 patients in the dasatinib 2L, and 207 in the nilotinib 2L cohorts. In 1L and 2L, infection-related HRU was higher for dasatinib than nilotinib cohorts. Infection-related inpatient (IP) days constituted a larger proportion of all-cause IP days in the 1L/2L dasatinib than 1L/2L nilotinib cohorts (dasatinib 1L/2L: 53%/58%; nilotinib 1L/2L: 50%/46%). Compared to the nilotinib cohort, the dasatinib cohort had higher all-cause total costs per patient per year by US$17,901 in 1L and $28,625 in 2L. Of the total cost difference, infection-related were $6048 (34%) in 1L and $28,192 (99%) in 2L, largely driven by IP cost differences (1L/2L: 96%/98%). Dasatinib was associated with higher HRU and healthcare costs compared to nilotinib, particularly related to infections. Novartis Pharmaceutical Corporation.


15-year outcome of treated bipolar disorder.
Prior reports suggested that bipolar patients in Taiwan had comparable long-term outcome to Western patients despite markedly lower rates of co-occurring substance use disorders. Thus, predictors of long-term outcome identified from Taiwanese bipolar samples may be less influenced by substance abuse. One hundred and one patients with bipolar disorder (DSM-III-R) having been naturalistically treated for at least 15 years were recruited. These patients were annually followed for 2 years to assess overall outcome, psychiatric symptoms, rehospitalization, work, and social adjustment. A combination of medical record reviews and direct personal interviews with patients and family members provided the clinical data. Of these patients, 16.8% expressed a poor overall long-term outcome, even though only two (2.0%) patients exhibited alcohol dependence during the follow-up period. Multivariate regression showed that full compliance with medication was the strongest predictor of favorable overall long-term outcome, followed by younger age at onset and male sex. Younger age at onset as well as male sex, but not full compliance, also predicted a favorable psychosocial outcome. Recruiting our sample from a clinical population with uncontrollable long-term treatment limits the generalizability of the findings. Compliance with pharmacotherapy is important to achieve a favorable overall long-term outcome of bipolar disorder. A portion of bipolar patients may have an unfavorable psychosocial outcome regardless of the psychopharmacological intervention or presence of substance abuse.


Genetically mediated brain abnormalities in schizophrenia.
Schizophrenia is a highly heritable, neurobehavioral disorder; however, the mode of inheritance is complex, and linkage findings have been difficult to replicate. Some consistent linkage findings have emerged on chromosomes 1, 6, 8, 11, 13, 15, and 22. New methods are being developed for candidate gene identification, including the use of neurobiologic phenotypes observed in relatives of persons with schizophrenia. Neuroimaging studies of relatives implicate abnormal hippocampal structure and inefficient prefrontal network functioning, probably representing mild variants of the abnormalities observed in schizophrenia. These characteristics may represent stable markers of vulnerability to schizophrenia, because they are not confounded by effects of antipsychotic drugs or psychosis. Recent studies provide evidence for a small role of the catechol-O-methyltransferase gene on 22q, and the serotonin receptor transporter gene on 17q11-q12 in the development of schizophrenia. Linking genes and brain regions or networks is an important step in identification of the pathophysiology of schizophrenia.


A medium-term corneal preserving medium (K-Sol).
A new corneal preserving solution, K-Sol, was prepared by combining TC 199, 100 mg/L of gentamicin sulfate, 0.025M HEPES buffer to a pH of 7.4, and various concentrations (2.0%; 2.25%; 2.5%) of chondroitin sulfate to a final osmolality of 310 mOsm. Rabbit corneas stored at 4 degrees C for 12 days in each solution, as well as fresh controls, were used for penetrating keratoplasty. Three weeks later, one graft in the 2.0% group was opacified; the rest were clear and showed minimal thickening. There were no significant differences between controls in terms of pachymetry and endothelial cell loss. This study supports the finding that K-Sol with 2.5% chondroitin sulfate is a simple, effective, medium-term preserving solution for donor tissue for penetrating keratoplasty in rabbits.


Event-driven architecture for health event detection from multiple sources.
Early detection of potential health threats is crucial for taking actions in time. It is unclear in which information source an event is reported first and, information from various sources can be complementing. Thus, it is important to search for information in a very broad range of sources. Furthermore, real-time processing is necessary to deal with the huge amounts of incoming data in time. Event-driven architectures are designed to address such challenges. This will be shown in this paper by presenting the architecture of a public health surveillance system that follows this style. Starting from concrete user requirements and scenarios, we introduce the architecture with its components for content collection, data analysis and integration. The system will allow for the monitoring of events in real-time as well as retrospectively.


Protection of Dairy Cattle in the EU: State of Play and Directions for Policymaking from a Legal and Animal Advocacy Perspective.
With the exception of a detailed Directive for calves, the welfare of dairy cattle is not regulated by species-specific legislation in the European Union. Their basic protection falls under the provisions of Directive 98/58/EC, also known as the "General Farm Animals Directive". Article 3 of this Directive states: "Member States shall make provision to ensure that the owners or keepers take all reasonable steps to ensure the welfare of animals under their care and to ensure that those animals are not caused any unnecessary pain, suffering or injury". However, recent reports show that the welfare of dairy cows in the EU is not sufficiently monitored and that serious problems persist. Lameness, mastitis, cubicle design, flooring, cleanliness, and permanent tethering remain critical areas. We argue that, to demonstrate compliance with Article 3 of Directive 98/58, farmers and Member States should urgently address these issues. The increasing proportion of cows that are never allowed to graze and high milk yields are also reasons for concern and will need to be addressed as a matter of priority in future EU guides to good practice and, eventually, legislation.


Does caring for your spouse harm one's health? Evidence from a United States nationally-representative sample of older adults.
The purpose of this article is to investigate the relationship between spousal care-giving and declines in functioning and self-rated health among older care-givers. The authors used data from the 2000 and 2002 waves of the United States Health and Retirement Study, a biennial longitudinal survey of a nationally representative cohort of adults aged 50 or more years. Two outcomes were examined, declines in functioning and declines in self-rated health. Care-givers were classified into three groups: no care-giving, less than 14 hours of care-giving per week, and 14 or more hours care-giving per week. To assess declines in functioning, two summary scores were created of limitations in basic and instrumental Activities of Daily Living. To assess declines in self-rated health, we compared responses from 2000 and 2002. In the fully adjusted models, care-giving hours did not have an independent effect on declines in functioning or self-rated health. The relationship between care-giving hours and declines in functioning and self-rated health is probably attributable to socio-demographic characteristics, mainly age. The findings suggest that spousal care-giving does not of itself harm functional health or perceived health among older adult care-givers. Understanding the differential effects of these socio-economic characteristics with care-giving hours on health will be useful in promoting the health of older adult care-givers and treating their disorders.


Use of real-time ultrasound during central venous catheter placement: Results of an APSA survey.
The purpose of this study was to document the attitudes and practice patterns of pediatric surgeons regarding use of RTUS with CVC placement. An analytic survey composed of 20 questions was sent via APSA headquarters to all practicing members. Answers were summarized as frequency and percentage. Distributions of answers were compared using the chi-square tests. P-values ≤0.05 were considered statistically significant. 361 of 1072 members chose to participate for a response rate of 34%. Most placed CVCs into the subclavian veins (SCV) of patients without coagulopathy, with the left SCV chosen approximately four times more often than the right. Conversely, RTUS use at the internal jugular vein (IJV) was significantly greater than that for the SCV (p<0.001). Coagulopathy, multiple previous catheters, and morbid obesity were identified as patient characteristics that would encourage RTUS use. The most commonly cited potential barriers to RTUS use were lack of formal ultrasound training and the belief that ultrasound is not necessary. Variability exists among pediatric surgeons regarding use of RTUS during CVC placement. Additional studies are needed to document actual frequency of use, how RTUS is implemented, and its efficacy of preventing adverse events in children.


Effects of water salinity on acid-base balance in decapod crustaceans.
Extracellular acid-base balance in decapod crustaceans is influenced by water salinity, although the nature of this relationship is unclear. In euryhaline crabs, a decrease in salinity results in a metabolic alkalosis in the haemolymph and an increase in salinity results in a metabolic acidosis. Alterations in acid-base status by external changes in salinity are thought to be secondary to the adjustments required for ionic and osmotic regulation. In the present study, acid-base adjustments in the haemolymph of Eriocheir sinensis after transfer to 30 % sea water accompanied alterations in muscle pH and [HCO(3)(-)], as an initial acidosis coincided with an alkalosis in the leg muscle. By 48 h transfer, haemolymph pH increased as muscle pH and HCO(3)(-) declined. Haemolymph [Cl(-)] decreased significantly 3 h after transfer to a new steady state but haemolymph [Na(+)] and muscle [Na(+)] and [Cl(-)] remained unchanged. Muscle free amino acid concentration increased twofold 6 h after transfer, followed by a 2.5-fold increase in the haemolymph after 24 h. In contrast, 30 % sea water had no effect on haemolymph acid-base adjustments in the osmoconforming crab, Necora puber, which lacks ion and osmo-regulatory mechansims. Collectively these observations support the view that salinity-induced alterations in acid-base status are caused by adjustments consistent with cell volume regulation.


Perturbation approach applied to modal diffraction methods.
Eigenvalue computation is an important part of many modal diffraction methods, including the rigorous coupled wave approach (RCWA) and the Chandezon method. This procedure is known to be computationally intensive, accounting for a large proportion of the overall run time. However, in many cases, eigenvalue information is already available from previous calculations. Some of the examples include adjacent slices in the RCWA, spectral- or angle-resolved scans in optical scatterometry and parameter derivatives in optimization. In this paper, we present a new technique that provides accurate and highly reliable solutions with significant improvements in computational time. The proposed method takes advantage of known eigensolution information and is based on perturbation method.


Distraction of eye-hand coordination varies with working memory capacity.
The authors present a study of the relationship between individual variation in working memory capacity (WMC) and visually guided hand control in the face of visual distraction. WMC was assessed with the automated operation span task. Hand control was measured by requesting participants to track a visual target with a hand-held touch screen pen. Tracking error increased when nontarget visual objects (distractors) appeared, especially in individuals with low WMC. High-WMC individuals are less impaired by distractors than their low-WMC counterpart, because they resume target tracking more quickly after distractor onset. The results suggest that visual distractors cause a momentary interruption to tracking movements and that high WMC attenuates this interruption by facilitating visual search.


Themis is a member of a new metazoan gene family and is required for the completion of thymocyte positive selection.
T cell antigen receptor (TCR) signaling in CD4(+)CD8(+) double-positive thymocytes determines cell survival and lineage commitment, but the genetic and molecular basis of this process is poorly defined. To address this issue, we used ethylnitrosourea mutagenesis to identify a previously unknown T lineage-specific gene, Themis, which is critical for the completion of positive selection. Themis contains a tandem repeat of a unique globular domain (called 'CABIT' here) that includes a cysteine motif that defines a family of five uncharacterized vertebrate proteins with orthologs in most animal species. Themis-deficient thymocytes showed no substantial impairment in early TCR signaling but did show altered expression of genes involved in the cell cycle and survival before and during positive selection. Our data suggest a unique function for Themis in sustaining positive selection.


Stress-induced eating in rats.
Mild tail pinch (TP) in rats resulted in 72% of animals displaying ingestive behavior with 20% demonstrating gnawing behavior without food ingestion and 8% demonstrating licking behavior only. The animals ate steadily over 5 min with a maximum rate occurring at 1 min (0.5 +/- 0.2 g). There was a circadian rhythm of TP-induced behavior with the peak food ingestion occurring at 24 h. A mild increase in blood glucose occurred 120 s after commencement of TP (115 +/- 4 mg/dl). Common satiety signals such as stomach distension and glucose decreased food ingestion. Parenteral administration of glucagon, cholecystokinin-octapeptide, bombesin, and thyrotropin-releasing hormone resulted in suppression of TP-induced food ingestion. Chronic TP (12 5-min TP periods/day) resulted in a fall in spontaneous food intake with the total intake remaining similar to food intake prior to the chronic TP period. We suggest that TP serves as an excellent model for eating behavior because 1) it correlates well with starvation-induced eating; 2) it precludes the necessary deprivation of food and water to adrenalectomized animals; and 3) animals subjected to TP continue chewing in the face of decreased food intake allowing one to exclude the possibility that the effects of an anorectic are secondary to nausea.


Calsequestrin 2 (CASQ2) mutations increase expression of calreticulin and ryanodine receptors, causing catecholaminergic polymorphic ventricular tachycardia.
Catecholamine-induced polymorphic ventricular tachycardia (CPVT) is a familial disorder caused by cardiac ryanodine receptor type 2 (RyR2) or calsequestrin 2 (CASQ2) gene mutations. To define how CASQ2 mutations cause CPVT, we produced and studied mice carrying a human D307H missense mutation (CASQ(307/307)) or a CASQ2-null mutation (CASQ(DeltaE9/DeltaE9)). Both CASQ2 mutations caused identical consequences. Young mutant mice had structurally normal hearts but stress-induced ventricular arrhythmias; aging produced cardiac hypertrophy and reduced contractile function. Mutant myocytes had reduced CASQ2 and increased calreticulin and RyR2 (with normal phosphorylated proportions) but unchanged calstabin levels, as well as reduced total sarcoplasmic reticulum (SR) Ca(2+), prolonged Ca(2+) release, and delayed Ca(2+) reuptake. Stress further diminished Ca(2+) transients, elevated cytosolic Ca(2+), and triggered frequent, spontaneous SR Ca(2+) release. Treatment with Mg(2+), a RyR2 inhibitor, normalized myocyte Ca(2+) cycling and decreased CPVT in mutant mice, indicating RyR2 dysfunction was critical to mutant CASQ2 pathophysiology. We conclude that CPVT-causing CASQ2 missense mutations function as null alleles. In the absence of CASQ2, calreticulin, a fetal Ca(2+)-binding protein normally downregulated at birth, remains a prominent SR component. Adaptive changes to CASQ2 deficiency (increased posttranscriptional expression of calreticulin and RyR2) maintained electrical-mechanical coupling, but increased RyR2 leakiness, a paradoxical response further exacerbated by stress. The central role of RyR2 dysfunction in CASQ2 deficiency unifies the pathophysiologic mechanism underlying CPVT due to RyR2 or CASQ2 mutations and suggests a therapeutic approach for these inherited cardiac arrhythmias.


The heterodimerization of platelet-derived chemokines.
Chemokines encompass a large family of proteins that act as chemoattractants and are involved in many biological processes. In particular, chemokines guide the migration of leukocytes during normal and inflammatory conditions. Recent studies reveal that the heterophilic interactions between chemokines significantly affect their biological activity, possibly representing a novel regulatory mechanism of the chemokine activities. The co-localization of platelet-derived chemokines in vivo allows them to interact. Here, we used nano-spray ionization mass spectrometry to screen eleven different CXC and CC platelet-derived chemokines for possible interactions with the two most abundant chemokines present in platelets, CXCL4 and CXCL7. Results indicate that many screened chemokines, although not all of them, form heterodimers with CXCL4 and/or CXCL7. In particular, a strong heterodimerization was observed between CXCL12 and CXCL4 or CXCL7. Compared to other chemokines, the main structural difference of CXCL12 is in the orientation and packing of the C-terminal alpha-helix in relation to the beta-sheet. The analysis of one possible structure of the CXCL4/CXCL12 heterodimer, CXC-type structure, using molecular dynamics (MD) trajectory reveals that CXCL4 may undergo a conformational transition to alter the alpha helix orientation. In this new orientation, the alpha-helix of CXCL4 aligns in parallel with the CXCL12 alpha-helix, an energetically more favorable conformation. Further, we determined that CXCL4 and CXCL12 physically interact to form heterodimers by co-immunoprecipitations from human platelets. Overall, our results highlight that many platelet-derived chemokines are capable of heterophilic interactions and strongly support future studies of the biological impact of these interactions.