Recruitment and recirculation of leukocytes after an ultramarathon run: preferential homing of cells expressing high levels of the adhesion molecule LFA-1.
The alpha L beta 2 (CD11a/CD18) integrin LFA-1 (lymphocyte function-associated antigen-1) mainly contributes to the firm arrest of leucocytes on the endothelium. Its cell surface density is thought to be important for the exercise-induced homing of leucocytes. 9 athletes (7 males, 2 females, age 36-68 years, body mass 64 +/- 10 kg, height: 175 +/- 10 cm) absolved a competitive 100 km run in 8:07 hours (range: 7:29-9:50 hours). Immunophenotyping of circulating leucocytes and data acquisition by three colour flow cytometry before and up to 3 hours after the race showed that post exercise lymphocyte subpopulations with a higher expression of CD11a like CD3+CD8+CD45RO+ T-cells, natural killer cells (CD3-CD16/CD56+) and cytotoxic, not MHC-restricted T-cells (CD3+CD16/CD56+) decreased significantly more than CD45RO+ and CD45RO- helper-T-cells (CD3+CD4+), CD3+CD8+CD45RO-T-cells and B-cells (CD19+), which have lower levels for CD11a. Cell concentrations of regular monocytes (Fc gamma receptor 3 negative, LFA-1 low positive) increased, whereas mature monocytes (Fc gamma receptor 3 low or high positive respectively, LFA-1 high positive) decreased. In conclusion the surface density of the adhesion molecule LFA-1 on leucocytes is likely to contribute significantly to the extent of leucocyte's homing after long endurance exercise.


Tbx18 and the fate of epicardial progenitors.
Uncovering the origins of myocardial cells is important for understanding and treating heart diseases. Cai et al. suggest that Tbx18-expressing epicardium provides a substantial contribution to myocytes in the ventricular septum and the atrial and ventricular walls. Here we show that the T-box transcription factor gene 18 (Tbx18) itself is expressed in the myocardium, showing that their genetic lineage tracing system does not allow conclusions of an epicardial origin of cardiomyocytes in vivo to be drawn.


Steroid sulfatase: a new target for the endocrine therapy of breast cancer.
Inhibitors of steroid sulfatase are being developed as a novel therapy for hormone-dependent breast cancer in postmenopausal women. Data suggest that steroid sulfatase (STS) activity is much higher than aromatase activity in breast tumors and high levels of STS mRNA expression in tumors are associated with a poor prognosis. STS hydrolyzes steroid sulfates, such as estrone sulfate and dehydroepiandrosterone sulfate (DHEAS), to estrone and DHEA, which can be converted to steroids with potent estrogenic properties, that is, estradiol and androstenediol, respectively. Several potent irreversible STS inhibitors have now been identified, including STX64 (BN83495), a tricyclic sulfamate ester. This drug recently completed the first-ever trial of this new type of therapy in postmenopausal women with estrogen receptor-positive metastatic breast cancer. STX64, tested at 5-mg and 20-mg doses, was able to almost completely block STS activity in peripheral blood lymphocytes and tumor tissues. Inhibition of STS activity was associated with significant reductions in serum concentrations of androstenediol and estrogens. Unexpectedly, serum androstenedione concentrations also decreased by up to 86%, showing that this steroid, which is the main substrate for the aromatase in postmenopausal women, is derived mainly from the peripheral conversion of DHEAS. Of eight patients who completed therapy, five showed evidence of stable disease for up to 7.0 months. This new endocrine therapy offers considerable potential for the treatment of hormone-dependent breast cancer in postmenopausal women.


Systematic identification of gene family regulators in mouse and human embryonic stem cells.
Pluripotent self-renewing embryonic stem cells (ESCs) have been the focus of a growing number of high-throughput experiments, revealing the genome-wide locations of hundreds of transcription factors and histone modifications. While most of these datasets were used in a specific context, all datasets combined offer a comprehensive view of chromatin characteristics and regulatory elements that govern cell states. Here, using hundreds of datasets in ESCs, we generated colocalization maps of chromatin proteins and modifications, and built a discovery pipeline for regulatory proteins of gene families. By comparing genome-wide binding data with over-expression and knockdown analysis of hundreds of genes, we discovered that the pluripotency-related factor NR5A2 separates mitochondrial from cytosolic ribosomal genes, regulating their expression. We further show that genes with a common chromatin profile are enriched for distinct Gene Ontology (GO) categories. Our approach can be generalized to reveal common regulators of any gene group; discover novel gene families, and identify common genomic elements based on shared chromatin features.


The socio-economic impact of important camel diseases as perceived by a pastoralist community in Kenya.
This paper presents the results of a study conducted in a pastoral community in Kenya using participatory appraisal approaches. The objective of the study was to assess the socio-economic impact of camel trypanosomosis (surra) according to the perceptions of the pastoralists. Four livestock grazing units were conveniently selected and in each of them, three groups of key informants comprising five to eight persons were selected for the participatory exercises. Five camel diseases were listed in order of importance according to their severity and frequency of occurrence including trypanosomosis, mange, non-specific diarrhoea, tick infestations and haemorrhagic septicaemia. The losses listed as incurred due to the five diseases were: losses in milk, meat, blood, fats and hides, dowry payments, and depreciation in sale of animals, losses due to infertility and abortions, and losses due to the cost of treatment. There was good agreement (P < 0.05) between the informant groups on the losses incurred as a result of the diseases for all the selected loss indicators. Surra and mange were given high median scores on all the indicators while non-specific diarrhoea, tick infestations, and haemorrhagic septicaemia received moderate median scores. Based on the study findings it is concluded that the camel plays a central role in the lives of Turkana pastoralists and that surra has a devastating social and economic impact. There is a need for veterinary and policy decision-makers to focus more attention on the control of surra in this arid and semi-arid area of Kenya.


Performance enhancements for GROMACS nonbonded interactions on BlueGene.
Several improvements to the previously optimized GROMACS BlueGene inner loops that evaluate nonbonded interactions in molecular dynamics simulations are presented. The new improvements yielded an 11% decrease in running time for both PME and other kinds of GROMACS simulations that use nonbonded table look-ups. Some other GROMACS simulations will show a small gain.


Modeling the electronic states and magnetic properties derived from the f1 configuration in lanthanocene and actinocene compounds.
The electronic structure and magnetic properties of a series of Kramers ions with f1 configuration in axial symmetry have been analyzed with a combination of theoretical methods: ab initio relativistic wavefunction methods as well as a crystal-field (CF) model with parameters extracted from the ab initio calculations. The molecules were treated in an idealized D8h and D7h symmetry and different combinations of active electrons and orbitals were used to describe correctly the nature of the ground and excited states. The spin Hamiltonian parameters for the Kramers doublet derived from the ground multiplet were determined and discussed on the basis of the crystal-field (CF) and spin-orbit interaction. The prime interests were the electron paramagnetic resonance g-factors and their relation to the complex geometry, crystal-field (CF) and spin-orbit coupling. The developed models agree well with the ab initio calculations and experimental reports.


Targeted disruption of the Wnk4 gene decreases phosphorylation of Na-Cl cotransporter, increases Na excretion and lowers blood pressure.
We recently generated Wnk4(D561A/+) knockin mice and found that a major pathogenesis of pseudohypoaldosteronism type II was the activation of the OSR1/SPAK kinase-NaCl cotransporter (NCC) phosphorylation cascade by the mutant WNK4. However, the physiological roles of wild-type WNK4 on the regulation of Na excretion and blood pressure, and whether wild-type WNK4 functions positively or negatively in this cascade, remained to be determined. In the present study, we generated WNK4 hypomorphic mice by deleting exon 7 of the Wnk4 gene. These mice did not show hypokalemia and metabolic alkalosis, but they did exhibit low blood pressure and increased Na and K excretion under low-salt diet. Phosphorylation of OSR1/SPAK and NCC was significantly reduced in the mutant mice as compared with their wild-type littermates. Protein levels of ROMK and Maxi K were not changed, but epithelial Na channel appeared to be activated as a compensatory mechanism for the reduced NCC function. Thus, wild-type WNK4 is a positive regulator for the WNK-OSR1/SPAK-NCC cascade, and WNK4 is a potential target of anti-hypertensive drugs.


Appetite change profiles in depression exhibit differential relationships between systemic inflammation and activity in reward and interoceptive neurocircuitry.
Appetite change is a defining feature of major depressive disorder (MDD), yet little neuroscientific evidence exists to explain why some individuals experience increased appetite when they become depressed while others experience decreased appetite. Previous research suggests depression-related appetite changes can be indicative of underlying neural and inflammatory differences among MDD subtypes. The present study explores the relationship between systemic inflammation and brain circuitry supporting food hedonics for individuals with MDD. Sixty-four participants (31 current, unmedicated MDD and 33 healthy controls [HC]) provided blood samples for analysis of an inflammatory marker, C-reactive protein (CRP), and completed a functional magnetic resonance imaging (fMRI) scan in which they rated the perceived pleasantness of various food stimuli. Random-effects multivariate modeling was used to explore group differences in the relationship between CRP and the coupling between brain activity and inferred food pleasantness (i.e., strength of the relationship between activity and pleasantness ratings). Results revealed that for MDD with increased appetite, higher CRP in blood related to greater coupling between orbitofrontal cortex and anterior insula activity and inferred food pleasantness. Compared to HC, all MDD exhibited a stronger positive association between CRP and coupling between activity in striatum and inferred food pleasantness. These findings suggest that for individuals with MDD, systemic low-grade inflammation is associated with differences in reward and interoceptive-related neural circuitry when making hedonic inferences about food stimuli. In sum, altered immunologic states may affect appetite and inferences about food reward in individuals with MDD and provide evidence for physiological subtypes of MDD.


Parvalbumin and GAD65 interneuron inhibition in the ventral hippocampus induces distinct behavioral deficits relevant to schizophrenia.
Hyperactivity within the ventral hippocampus (vHPC) has been linked to both psychosis in humans and behavioral deficits in animal models of schizophrenia. A local decrease in GABA-mediated inhibition, particularly involving parvalbumin (PV)-expressing GABA neurons, has been proposed as a key mechanism underlying this hyperactive state. However, direct evidence is lacking for a causal role of vHPC GABA neurons in behaviors associated with schizophrenia. Here, we probed the behavioral function of two different but overlapping populations of vHPC GABA neurons that express either PV or GAD65 by selectively inhibiting these neurons with the pharmacogenetic neuromodulator hM4D. We show that acute inhibition of vHPC GABA neurons in adult mice results in behavioral changes relevant to schizophrenia. Inhibiting either PV or GAD65 neurons produced distinct behavioral deficits. Inhibition of PV neurons, affecting ∼80% of the PV neuron population, robustly impaired prepulse inhibition of the acoustic startle reflex (PPI), startle reactivity, and spontaneous alternation, but did not affect locomotor activity. In contrast, inhibiting a heterogeneous population of GAD65 neurons, affecting ∼40% of PV neurons and 65% of cholecystokinin neurons, increased spontaneous and amphetamine-induced locomotor activity and reduced spontaneous alternation, but did not alter PPI. Inhibition of PV or GAD65 neurons also produced distinct changes in network oscillatory activity in the vHPC in vivo. Together, these findings establish a causal role for vHPC GABA neurons in controlling behaviors relevant to schizophrenia and suggest a functional dissociation between the GABAergic mechanisms involved in hippocampal modulation of sensorimotor processes.


Is the devil in the detail? A randomised controlled trial of guided internet-based CBT for perfectionism.
An internet guided self-help cognitive-behavioural treatment (ICBT) for perfectionism was recently found to be effective (see this issue). Such studies stand in need of replication. The aim of this study was to report the outcomes and predictors of change when the treatment is delivered in a UK setting. A total of 120 people (Mean = 28.9 years; 79% female) were randomised to receive ICBT or wait-list control over 12 weeks (trial registration: NCT02756871). While there were strong similarities between the current study and its Swedish counterpart, there were also important differences in procedural details. There was a significant impact of the intervention on the primary outcome measure (Frost Multidimensional Perfectionism Scale, Concern over Mistakes subscale) and also on the Clinical Perfectionism Questionnaire (between group effect sizes d = 0.98 (95% CI: 0.60-1.36) and d = 1.04 (95% CI: 0.66-1.43) respectively using intent-to-treat analyses). Unlike the Swedish study, there was significant non-engagement and non-completion of modules with 71% of participants completing fewer than half the modules. The number of modules completed moderated the rate of change in clinical perfectionism over time. In conclusion, the study indicates the intervention is effective in a UK setting but highlighted the importance of procedural details to optimise retention.


Case report: recovery from persistent septicemic melioidosis.
Septicemic melioidosis is often fatal despite treatment with antibiotics such as ceftazidime to which Burkholderia pseudomallei, the causal pathogen, is sensitive in vitro. We report a near-fatal case of septicemic melioidosis with persistent B. pseudomallei bacteremia despite intravenous ceftazidime in which combination therapy with meropenem and ciprofloxacin, splenectomy and correction of metabolic acidosis allowed for hospital discharge. The choice of antibiotic agents was supported by intracellular minimum inhibitory concentration analysis using B. pseudomallei co-culture in Acanthamoeba trophozoites. The patient's B. pseudomallei isolates were indistinguishable by pulsed-field gel electrophoresis from clinical and environmental isolates previously analyzed during investigation of a Western Australian melioidosis outbreak. A combination of antibiotics known to possess intracellular activity against B. pseudomallei, surgery and supportive critical care may provide a means of improving the probability of survival in persistent septicemic melioidosis.


Photoperiod controls vegetation phenology across Africa.
Vegetation phenology is driven by environmental factors such as photoperiod, precipitation, temperature, insolation, and nutrient availability. However, across Africa, there's ambiguity about these drivers, which can lead to uncertainty in the predictions of global warming impacts on terrestrial ecosystems and their representation in dynamic vegetation models. Using satellite data, we undertook a systematic analysis of the relationship between phenological parameters and these drivers. The analysis across different regions consistently revealed photoperiod as the dominant factor controlling the onset and end of vegetation growing season. Moreover, the results suggest that not one, but a combination of drivers control phenological events. Consequently, to enhance our predictions of climate change impacts, the role of photoperiod should be incorporated into vegetation-climate and ecosystem modelling. Furthermore, it is necessary to define clearly the responses of vegetation to interactions between a consistent photoperiod cue and inter-annual variation in other drivers, especially under a changing climate.


Clinical and serological autoimmune complications in chronic lymphocytic leukemia.
Autoimmune disorders often develop during the course of chronic lymphocytic leukemia (CLL). The aim of our study was to investigate the incidence of autoimmune complications (AIC) and serological autoantibodies, and to assess the relationship of these to patient characteristics. We prospectively collected screenings of AIC and serological markers from a total of 192 patients. AIC was observed in 18 (9.4%) patients. Autoimmune hemolytic anemia (AIHA) was observed in 8 patients. Autoimmune thrombocytopenia (AITP) was observed in 3 patients. Other various types of AIC were observed in the remaining 7 patients. Serological autoantibodies were positive in 17.2% of patients with CLL. The mean age of patients with AIC was higher than the control group (p = 0.036). Patients with AIC were mostly in advanced disease stage (p = 0.004), and they had received more first-line treatments than the control group (p = 0.003). Patients with AIC had a higher mean age and more advanced disease stage than patients with positive serological autoantibodies (p = 0.020 and p = 0.009; respectively). In addition, patients with AIC had also received more first-line treatment than the patients with positive serological autoantibodies (p = 0.015). Hematologic AIC was associated with older age, advanced disease stage, and treatment. Conversely, non-hematological AIC and serological autoantibodies are generally observed in early stages. Our study has established a coexistence of CLL and autoimmune complications. Hematologists are usually familiar with AIHA and AITP, but less so with non-hematologic AIC. The latter complications should be carefully searched for, particularly in patients with early CLL.


Measurable Residual Disease at Induction Redefines Partial Response in Acute Myeloid Leukemia and Stratifies Outcomes in Patients at Standard Risk Without NPM1 Mutations.
Purpose We investigated the effect on outcome of measurable or minimal residual disease (MRD) status after each induction course to evaluate the extent of its predictive value for acute myeloid leukemia (AML) risk groups, including NPM1 wild-type (wt) standard risk, when incorporated with other induction response criteria. Methods As part of the NCRI AML17 trial, 2,450 younger adult patients with AML or high-risk myelodysplastic syndrome had prospective multiparameter flow cytometric MRD (MFC-MRD) assessment. After course 1 (C1), responses were categorized as resistant disease (RD), partial remission (PR), and complete remission (CR) or complete remission with absolute neutrophil count < 1,000/µL or thrombocytopenia < 100,000/μL (CRi) by clinicians, with CR/CRi subdivided by MFC-MRD assay into MRD+ and MRD-. Patients without high-risk factors, including Flt3 internal tandem duplication wt/- NPM1-wt subgroup, received a second daunorubicin/cytosine arabinoside induction; course 2 (C2) was intensified for patients with high-risk factors. Results Survival outcomes from PR and MRD+ responses after C1 were similar, particularly for good- to standard-risk subgroups (5-year overall survival [OS], 27% RD v 46% PR v 51% MRD+ v 70% MRD-; P < .001). Adjusted analyses confirmed significant OS differences between C1 RD versus PR/MRD+ but not PR versus MRD+. CRi after C1 reduced OS in MRD+ (19% CRi v 45% CR; P = .001) patients, with a smaller effect after C2. The prognostic effect of C2 MFC-MRD status (relapse: hazard ratio [HR], 1.88 [95% CI, 1.50 to 2.36], P < .001; survival: HR, 1.77 [95% CI, 1.41 to 2.22], P < .001) remained significant when adjusting for C1 response. MRD positivity appeared less discriminatory in poor-risk patients by stratified analyses. For the NPM1-wt standard-risk subgroup, C2 MRD+ was significantly associated with poorer outcomes (OS, 33% v 63% MRD-, P = .003; relapse incidence, 89% when MRD+ ≥ 0.1%); transplant benefit was more apparent in patients with MRD+ (HR, 0.72; 95% CI, 0.31 to 1.69) than those with MRD- (HR, 1.68 [95% CI, 0.75 to 3.85]; P = .16 for interaction). Conclusion MFC-MRD can improve outcome stratification by extending the definition of partial response after first induction and may help predict NPM1-wt standard-risk patients with poor outcome who benefit from transplant in the first CR.


Role of immunomarkers in the clinicopathological analysis of unicystic ameloblastoma.
The clinical behavior of unicystic ameloblastoma varies according to its subtype. The assessment of its proliferative capacity, neovascularization, and invasiveness using relevant immunomarkers may aid in appropriate surgical therapeutic protocol. 18 cases of clinically and histologically confirmed unicystic ameloblastoma, categorized as luminal, intraluminal, or mural subtypes, were analyzed retrospectively. Immunomarkers such as Ki-67, CD34, MMP-2, and MMP-9 were studied to evaluate their behavior. Labeling index of Ki-67 was 4.25% in the intraluminal subtype, compared with 2.14% in the luminal and 4.04% in the mural variant (P = 0.3). CD34 immunostaining was significantly higher in the mural variant (43 per high power field) than the other two subtypes (P = 0.04). MMP-2 and MMP-9 were strongly expressed in mural, moderately in intraluminal, and weakly to absent in luminal variant. High proliferative index, angiogenesis, and protease activity in the mural ameloblastoma, ascertained by the expression of these markers, confirm its aggressive phenotype. The intraluminal and luminal subtype exhibiting decreased expression are compatible with their indolent clinical behavior.


Comparative validity of methods to select appropriate cutoff weight for probabilistic linkage without unique personal identifiers.
Record linkage can enhance data quality of observational database studies. Probabilistic linkage, a method that allows partial match of linkage variables, overcomes disagreements arising from errors and omissions in data entry but also results in false-positive links. The study aimed to assess the validity of probabilistic linkage in the absence of unique personal identifiers (UPI) and the methods of cutoff weight selection. We linked an implantable cardioverter defibrillator placement registry to Medicare inpatient files of 1 year with anonymous nonunique variables and assessed the validity of three methods of cutoff selection against an internally derived gold standard with UPI. Of the 64,890 registry records with an expected linkage rate of 55-65%, 55% were linked at cutoffs associated with positive predictive value (PPV) of ≥90%. Histogram inspection suggested an approximate range of optimal cutoffs. The duplicate method made accurate estimates of cutoff and PPV if the method's assumption was met. With adjusted estimates of the sizes of true matches and searched files, the odds formula method made relatively accurate estimates of cutoff and PPV. Probabilistic linkage without UPI generated valid linkages when an optimal cutoff was chosen. Cutoff selection remains challenging; however, histogram inspection, the duplicate method, and the odds formula method can be used in conjunction when a gold standard is not available.


Complete objective response of neuroblastoma to biological treatment.
The combined use (MDB) of Somatostatin, Melatonin, Retinoids, Vitamins E, C, and D3, with Calcium, Chondroitin sulfate, and microdoses of Cyclophosphamide in a seven-month old baby affected by a voluminous retroperitoneal neuroblastoma measuring 4 x 8 cm produced a 50% objective response in six months, an almost total response in one year and a complete response at 14 months, with cure and absence of disease for over ten years. This paper discusses the rationale and the molecular mechanisms of action of the treatment which has a differentiating, apoptotic and antiproliferative effect, preserving and enhancing both the trophism and functionality of organs and tissues, and the neuroimmunoendocrine and antiblastic homeostasis. This result is in agreement with the positive results already published on the use of the MDB in lymphoproliferative diseases, in stage III and IV lung cancer, in breast cancer and in cancers of the upper aerodigestive epithelia. Without the need for hospitalisation and with no toxicity, the MDB rapidly reduced and then eliminated the voluminous tumoral mass, allowing a normal quality of life and a perfectly normal psychophysical development. We believe it is of use to report this case in order to invite greater interest in the oncological possibilities offered by the immunoneuroendocrine and biological-receptorial properties of the MDB treatment.


Robust and distributed neural representation of action values.
Studies in rats, monkeys, and humans have found action-value signals in multiple regions of the brain. These findings suggest that action-value signals encoded in these brain structures bias choices toward higher expected rewards. However, previous estimates of action-value signals might have been inflated by serial correlations in neural activity and also by activity related to other decision variables. Here, we applied several statistical tests based on permutation and surrogate data to analyze neural activity recorded from the striatum, frontal cortex, and hippocampus. The results show that previously identified action-value signals in these brain areas cannot be entirely accounted for by concurrent serial correlations in neural activity and action value. We also found that neural activity related to action value is intermixed with signals related to other decision variables. Our findings provide strong evidence for broadly distributed neural signals related to action value throughout the brain.


Transcriptional control of the human glucocorticoid receptor: identification and analysis of alternative promoter regions.
Glucocorticoid receptor levels are thought to be controlled by multiple alternative first exons. Seven of these exons are located in an upstream CpG island. In this study, we investigated the promoter activity of the intronic regions between these exons, and their susceptibility to CpG methylation and sequence variability. The seven promoters were cloned into luciferase reporter genes, and their activity measured in ten cell lines. CpG islands of 221 donors were genotyped and the effects of these SNPs were investigated in a reporter gene assay. We showed that each of the first exons was independently controlled by a unique promoter located directly upstream. Promoter activities were cell type-specific, and varied considerably between cell types. Irrespective of the cell type, in vitro methylation effectively silenced all reporter constructs. Eleven SNPs were observed within the CpG island of 221 donors, and a new promoter-specific haplotype was revealed. Four of the minor alleles reduced the reporter gene activity, with cell type specific effects. This complexity within the CpG island helps to explain the variable, tissue-specific transcriptional control of the GR, and provides insight into the mechanisms underlying tissue specific deregulation of GR levels.


Efficacy of Epley Maneuver on Quality of Life of Elderly Patients with Subjective BPPV.
This study aimed to evaluate the efficacy of the repositioning maneuver on quality of life in elderly patients with dizziness and/or vertigo. This controlled, prospective randomized clinical trial was conducted in elderly patients aged 65 years and above with a positive history of benign paroxysmal positional vertigo (BPPV), presence of vertigo, and no observable nystagmus during the Dix-Hallpike test, so-called Subjective BPPV (S-BPPV). Individuals were evaluated by visual analog scale (VAS) and dizziness handicap inventory (DHI). Groups were defined as treatment (treated with Epley maneuver bilaterally) or no treatment control (no treatment modality or canalith repositioning maneuver). Ten days after the first assessment, all patients were reassessed using VAS and DHI. A total of 50 patients were randomized into two groups: 25 to the treatment group, and 25 to the control group. No significant differences were observed for baseline VAS and total DHI scores between the groups (p=0.636, p=0.846, respectively). On the other hand, after the reassessment, VAS and total DHI scores were both significantly reduced in the treatment group (p<0.001, p<0.001, respectively), but no reduction in either score was found in the control group (p=0.216, p=0.731, respectively). This study showed that elderly patients with S-BPPV benefit from the Epley maneuver, in particular global and disease-specific quality of life.


High-level production of alpha-particle-emitting (211)At and preparation of (211)At-labeled antibodies for clinical use.
In vitro and in vivo studies in human glioma models suggest that the antitenascin monoclonal antibody 81C6 labeled with the 7.2-h-half-life alpha-particle emitter (211)At might be a valuable endoradiotherapeutic agent for the treatment of brain tumors. The purpose of this study was to develop methods for the production of high levels of (211)At and the radiosynthesis of clinically useful amounts of (211)At-labeled human/mouse chimeric 81C6 antibody. (211)At was produced through the (209)Bi(alpha, 2n)(211)At reaction using an internal target system and purified by a dry distillation process. Antibody labeling was accomplished by first synthesizing N-succinimidyl 3-[(211)At]astatobenzoate from the corresponding tri-n-butyl tin precursor and reacting it with the antibody in pH 8.5 borate buffer. Quality control procedures consisted of methanol precipitation, size-exclusion high-performance liquid chromatography (HPLC), and pyrogen and sterility assays, as well as determination of the immunoreactive fraction by a rapid procedure using a recombinant tenascin fragment coupled to magnetic beads. A total of 16 antibody labeling runs were performed. Using beam currents of 50-60 microA alpha-particles and irradiation times of 1.5-4.5 h, the mean (211)At production yield was 27.75 +/- 2.59 MBq/microA.h, and the maximum level of (211)At produced was 6.59 GBq after a 4-h irradiation at 55 microA. The decay-corrected distillation yield was 67% +/- 16%. The yield for the coupling of the (211)At-labeled active ester to the antibody was 76% +/- 8%. The fraction of (211)At activity that eluted with a retention time corresponding to intact IgG on HPLC was 96.0% +/- 2.5%. All preparations had a pyrogen level of <0.125 EU/mL and were determined to be sterile. The mean immunoreactive fraction for these 16 preparations was 83.3% +/- 5.3%. Radiolysis did not interfere with labeling chemistry or the quality of the labeled antibody product. These results show that it is feasible to produce clinically relevant activities of (211)At-labeled antibodies and have permitted the initiation of a phase I trial of (211)At-labeled chimeric 81C6 administered directly into the tumor resection cavities of brain tumor patients.


Realization of masticatory movement by 3-dimensional simulation of the temporomandibular joint and the masticatory muscles.
Masticatory muscles are closely involved in mastication, pronunciation, and swallowing, and it is therefore important to study the specific functions and dynamics of the mandibular and masticatory muscles. However, the shortness of muscle fibers and the diversity of movement directions make it difficult to study and simplify the dynamics of mastication. The purpose of this study was to use 3-dimensional (3D) simulation to observe the functions and movements of each of the masticatory muscles and the mandible while chewing. To simulate the masticatory movement, computed tomographic images were taken from a single Korean volunteer (30-year-old man), and skull image data were reconstructed in 3D (Mimics; Materialise, Leuven, Belgium). The 3D-reconstructed masticatory muscles were then attached to the 3D skull model. The masticatory movements were animated using Maya (Autodesk, San Rafael, CA) based on the mandibular motion path. During unilateral chewing, the mandible was found to move laterally toward the functional side by contracting the contralateral lateral pterygoid and ipsilateral temporalis muscles. During the initial mouth opening, only hinge movement was observed at the temporomandibular joint. During this period, the entire mandible rotated approximately 13 degrees toward the bicondylar horizontal plane. Continued movement of the mandible to full mouth opening occurred simultaneously with sliding and hinge movements, and the mandible rotated approximately 17 degrees toward the center of the mandibular ramus. The described approach can yield data for use in face animation and other simulation systems and for elucidating the functional components related to contraction and relaxation of muscles during mastication.


Symptomatic cholecystolithiasis after cholecystectomy.
A 43-year-old woman was admitted to the gastroenterology department with colicky pain in the upper abdomen. Four years earlier, she had undergone a laparoscopic cholecystectomy because of cholecystitis. She recognised her current complaints from that previous episode. An endoscopic retrograde cholangiopancreatography showed a cavity with a diameter of 2 cm which contained multiple concrements near the liver hilus. An elective surgical exploration was performed. Near the clip of the previous cholecystectomy a bulging of the biliary tract with its own duct was visualised and resected. Histological examination of this "neo" gallbladder showed that the bulging was consistent with the formation of a reservoir secondary to bile leakage, probably caused by a small peroperative lesion of the common bile duct during the previous cholecystectomy. In conclusion, our patient presented with colicky pain caused by concrements inside a 'neo' gallbladder.


glnA mutations conferring resistance to methylammonium in Escherichia coli K12.
Cells of Escherichia coli K12 were sensitive to 100 mM-methylammonium when cultured under nitrogen limitation, and resistant when grown with an excess of either NH4Cl or glutamine. Glutamine synthetase activity was required for expression of the methylammonium-sensitive phenotype. Mutants were isolated which were resistant to 100 mM-methylammonium, even when grown under nitrogen limitation. P1 bacteriophage transduction and F' complementation analysis revealed that the resistance-conferring mutations mapped either inside the glnA structural gene and/or elsewhere in the E. coli chromosome. Glutamine synthetase was purified from the wild-type and from some of the mutant strains. Strains carrying glnA-linked mutations that were solely responsible for the methylammonium-resistant phenotype yielded an altered enzyme, which was less active biosynthetically with either ammonium or methylammonium as substrate. Sensitivity to methylammonium appeared to be due to synthesis of gamma-glutamylmethylamide by glutamine synthetase, which was synthesized poorly, if at all, by mutants carrying an altered glutamine synthetase enzyme.


SU-E-T-472: Characterization of the Very High Energy Electrons, ISO - 250 MeV (VHEE) Beam Generated by ALPHA-X Laser Wakefield Accelerator Beam Line for Utilization in Monte Carlo Simulation for Biomedical Experiment Planning.
Progress in the development of compact high-energy pulsed laser- plasma wakefield accelerators is opening up the potential for using Very High Energy Electron (VHEEs) beams in the range of 150 - 250 MeV for biomedical studies. Initial experiments using VHEE for this purpose have been carried out using the ALPHA-X laser-plasma wakefield accelerator beam line at the University of Strathclyde, Glasgow, UK. The purpose of this investigation is to use Monte Carlo simulations to plan experiments and compare with characterization of the interaction of the VHEE beam using a dosimeter. An experiment using the VHEE beam to irradiate a muscle-equivalent BANG polymer gel dosimeter has been carried out. Simulations have been used to prepare for the experiments. These were undertaken using the expected average energy for a pulse set and an energy spread approximated by Gaussian distribution. The model was implemented in FLUKA Monte Carlo code with follow up modeling using the Geant4 toolkit. The results have been compared with 1mm̂3 voxel laser CT based measurements of the dose deposited in the BANG dosimeter and with measurement of the induced radioactivity. The results of the measured dose from induced radioactivity have been compared with data from the FLUKA simulations. The beam model based on an average energy of particles in irradiation gives an acceptable estimate of the induced radioactivity and the dose deposited in the BANG dosimeter. Comparison with the dosimeter scanned profiles shows that the structure of the spectra of VHEE beams in the experiment and secondary scattered particles in the beam line should be accounted for in any model. Such model description of the VHEE beam for the ALPHA-X beam line has been developed. Monte Carlo simulations using the FLUKA code is an efficient way to plan a VHEE experiment and analyze data from measurements.


The intracellular domain of β-dystroglycan mediates the nucleolar stress response by suppressing UBF transcriptional activity.
β-dystroglycan (β-DG) is a key component of multiprotein complexes in the plasma membrane and nuclear envelope. In addition, β-DG undergoes two successive proteolytic cleavages that result in the liberation of its intracellular domain (ICD) into the cytosol and nucleus. However, stimuli-inducing ICD cleavage and the physiological relevance of this proteolytic fragment are largely unknown. In this study we show for the first time that β-DG ICD is targeted to the nucleolus where it interacts with the nuclear proteins B23 and UBF (central factor of Pol I-mediated rRNA gene transcription) and binds to rDNA promoter regions. Interestingly DG silencing results in reduced B23 and UBF levels and aberrant nucleolar morphology. Furthermore, β-DG ICD cleavage is induced by different nucleolar stressors, including oxidative stress, acidosis, and UV irradiation, which implies its participation in the response to nucleolar stress. Consistent with this idea, overexpression of β-DG elicited mislocalization and decreased levels of UBF and suppression of rRNA expression, which in turn provoked altered ribosome profiling and decreased cell growth. Collectively our data reveal that β-DG ICD acts as negative regulator of rDNA transcription by impeding the transcriptional activity of UBF, as a part of the protective mechanism activated in response to nucleolar stress.


Drug-loading capacity and nuclear targeting of multiwalled carbon nanotubes grafted with anionic amphiphilic copolymers.
In this study, three types of hybrid nanotubes (NTs), ie, oxidized multiwalled carbon NTs (COOH MWCNTs), heparin (Hep)-conjugated MWCNTs (Hep MWCNTs), and diblock copolymer polyglycolic acid (PGA)-co-heparin conjugated to MWCNTs (PGA MWCNTs), were synthesized with improved biocompatibility and drug-loading capacity. Hydrophilic Hep substituents on MWCNTs improved biocompatibility and acted as nucleus-sensitive segments on the CNT carrier, whereas the addition of PGA enhanced drug-loading capacity. In the PGA MWCNT system, the amphiphilic copolymer (PGA-Hep) formed micelles on the side walls of CNTs, as confirmed by electron microscopy. The PGA system encapsulated the hydrophobic drug with high efficiency compared to the COOH MWCNT and Hep MWCNT systems. This is because the drug was loaded onto the PGA MWCNTs through hydrophobic forces and onto the CNTs by π-π stacking interactions. Additionally, most of the current drug-carrier designs that target cancer cells release the drug in the lysosome or cytoplasm. However, nuclear-targeted drug release is expected to kill cancer cells more directly and efficiently. In our study, PGA MWCNT carriers effectively delivered the active anticancer drug doxorubicin into targeted nuclei. This study may provide an effective strategy for the development of carbon-based drug carriers for nuclear-targeted drug delivery.


High levels of fluctuating asymmetry in isolated stickleback populations.
Fluctuating asymmetry (FA), defined as small random deviations from the ideal bilateral symmetry, has been hypothesized to increase in response to both genetic and environmental stress experienced by a population. We compared levels of FA in 12 bilateral meristic traits (viz. lateral-line system neuromasts and lateral plates), and heterozygosity in 23 microsatellite loci, among four marine (high piscine predation risk) and four pond (zero piscine predation risk) populations of nine-spined sticklebacks (Pungitius pungitius). Pond sticklebacks had on average three times higher levels of FA than marine fish and this difference was highly significant. Heterozygosity in microsatellite markers was on average two times lower in pond (HE ≈ 0.3) than in marine (HE ≈ 0.6) populations, and levels of FA and heterozygosity were negatively correlated across populations. However, after controlling for habitat effect on heterozygosity, levels of FA and heterozygosity were uncorrelated. The fact that levels of FA in traits likely to be important in the context of predator evasion were elevated in ponds compared to marine populations suggests that relaxed selection for homeostasis in ponds lacking predatory fish may be responsible for the observed habitat difference in levels of FA. This inference also aligns with the observation that the levels of genetic variability across the populations did not explain population differences in levels of FA after correcting for habitat effect. Hence, while differences in strength of selection, rather than in the degree of genetic stress could be argued to explain habitat differences in levels of FA, the hypothesis that increased FA in ponds is caused by genetic stress cannot be rejected.


Function and dysfunction of CNG channels: insights from channelopathies and mouse models.
Channels directly gated by cyclic nucleotides (CNG channels) are important cellular switches that mediate influx of Na+ and Ca2+ in response to increases in the intracellular concentration of cAMP and cGMP. In photoreceptors and olfactory receptor neurons, these channels serve as final targets for cGMP and cAMP signaling pathways that are initiated by the absorption of photons and the binding of odorants, respectively. CNG channels have been also found in other types of neurons and in non-excitable cells. However, in most of these cells, the physiological role of CNG channels has yet to be determined. CNG channels have a complex heteromeric structure. The properties of individual subunits that assemble in specific stoichiometries to the native channels have been extensively investigated in heterologous expression systems. Recently, mutations in human CNG channel genes leading to inherited diseases (so-called channelopathies) have been functionally characterized. Moreover, mouse knockout models were generated to define the role of CNG channel proteins in vivo. In this review, we will summarize recent insights into the physiological and pathophysiological role of CNG channel proteins that have emerged from genetic studies in mice and humans.


Short Maternal Stature Increases Risk of Small-for-Gestational-Age and Preterm Births in Low- and Middle-Income Countries: Individual Participant Data Meta-Analysis and Population Attributable Fraction.
Small-for-gestational-age (SGA) and preterm births are associated with adverse health consequences, including neonatal and infant mortality, childhood undernutrition, and adulthood chronic disease. The specific aims of this study were to estimate the association between short maternal stature and outcomes of SGA alone, preterm birth alone, or both, and to calculate the population attributable fraction of SGA and preterm birth associated with short maternal stature. We conducted an individual participant data meta-analysis with the use of data sets from 12 population-based cohort studies and the WHO Global Survey on Maternal and Perinatal Health (13 of 24 available data sets used) from low- and middle-income countries (LMIC). We included those with weight taken within 72 h of birth, gestational age, and maternal height data (n = 177,000). For each of these studies, we individually calculated RRs between height exposure categories of < 145 cm, 145 to < 150 cm, and 150 to < 155 cm (reference: ≥ 155 cm) and outcomes of SGA, preterm birth, and their combination categories. SGA was defined with the use of both the International Fetal and Newborn Growth Consortium for the 21st Century (INTERGROWTH-21st) birth weight standard and the 1991 US birth weight reference. The associations were then meta-analyzed. All short stature categories were statistically significantly associated with term SGA, preterm appropriate-for-gestational-age (AGA), and preterm SGA births (reference: term AGA). When using the INTERGROWTH-21st standard to define SGA, women < 145 cm had the highest adjusted risk ratios (aRRs) (term SGA-aRR: 2.03; 95% CI: 1.76, 2.35; preterm AGA-aRR: 1.45; 95% CI: 1.26, 1.66; preterm SGA-aRR: 2.13; 95% CI: 1.42, 3.21). Similar associations were seen for SGA defined by the US reference. Annually, 5.5 million term SGA (18.6% of the global total), 550,800 preterm AGA (5.0% of the global total), and 458,000 preterm SGA (16.5% of the global total) births may be associated with maternal short stature. Approximately 6.5 million SGA and/or preterm births in LMIC may be associated with short maternal stature annually. A reduction in this burden requires primary prevention of SGA, improvement in postnatal growth through early childhood, and possibly further intervention in late childhood and adolescence. It is vital for researchers to broaden the evidence base for addressing chronic malnutrition through multiple life stages, and for program implementers to explore effective, sustainable ways of reaching the most vulnerable populations.


Rare germline copy number variants (CNVs) and breast cancer risk.
Germline copy number variants (CNVs) are pervasive in the human genome but potential disease associations with rare CNVs have not been comprehensively assessed in large datasets. We analysed rare CNVs in genes and non-coding regions for 86,788 breast cancer cases and 76,122 controls of European ancestry with genome-wide array data. Gene burden tests detected the strongest association for deletions in BRCA1 (P = 3.7E-18). Nine other genes were associated with a p-value < 0.01 including known susceptibility genes CHEK2 (P = 0.0008), ATM (P = 0.002) and BRCA2 (P = 0.008). Outside the known genes we detected associations with p-values < 0.001 for either overall or subtype-specific breast cancer at nine deletion regions and four duplication regions. Three of the deletion regions were in established common susceptibility loci. To the best of our knowledge, this is the first genome-wide analysis of rare CNVs in a large breast cancer case-control dataset. We detected associations with exonic deletions in established breast cancer susceptibility genes. We also detected suggestive associations with non-coding CNVs in known and novel loci with large effects sizes. Larger sample sizes will be required to reach robust levels of statistical significance.


Absence of dengue 2 infection enhancement in human sera containing Japanese encephalitis antibodies.
Sera from 52 young adults resident in a rural area in North Thailand were studied for plaque-reducing neutralizing antibodies against dengue (DEN) viruses types 1-4 and Japanese encephalitis (JE), and for DEN-2 infection-enhancing antibodies using a newly described microtest in the human monocyte cell line, U-937. Infection-enhancing antibody titers in U-937 cells using a simplified micromethod were similar to results obtained by published methods using human peripheral blood leukocytes and a macrotest using U-937 cells. In the sample, there were 23 with antibodies to one or more DEN viruses with or without accompanying JE antibodies; 16 sera demonstrated antibodies only to JE and 13 had no detectable antibodies to any flavivirus. All but two DEN antibody-containing sera enhanced DEN-2 infections in U-937 cells, often to titers of 1:10,000 or greater. By contrast, only one of 16 JE-immune sera enhanced DEN-2 infection in monocytes, and that at a dilution of 1:100. None of the flavivirus-negative sera had DEN-2 enhancing activity. The failure of human anti-JE contrasts with the ability of rabbit anti-JE to enhance DEN-2 infections, but correlates with the absence of recorded instances of dengue shock syndrome in human beings sequentially infected with JE and then a DEN virus. This report seemingly reconciles in vitro and in vivo phenomena, and may provide an opportunity to study mechanisms involved.


A systematic review of outbreak and non-outbreak studies of extraintestinal pathogenic Escherichia coli causing community-acquired infections.
A systematic review of outbreak and non-outbreak studies of infections caused by extraintestinal pathogenic Escherichia coli (ExPEC) was conducted. This review examines the epidemiology, seasonality, source or mode of transmission, and temporal changes, based on E. coli serogroup, in ExPEC causing sporadic vs. outbreak-associated infections. Twelve outbreak and 28 non-outbreak studies were identified. The existence of ExPEC outbreaks was well supported. Three of four outbreak reports indicated peak periods during the winter months. Serogroups associated with outbreak infections ranged from 1% to 26% (average 11·4%) vs. (range 1-15%, average 3·5%) for serogroups associated with sporadic infections; the distribution of serogroups also differed for outbreak and non-outbreak infections. Study authors indicated that the outbreaks may have resulted from foodborne transmission, but direct evidence was unavailable. This review provides evidence that the epidemiology of endemic vs. epidemic ExPEC infections differs; however, study reporting quality limited epidemiological inferences.


Reducing liver cancer disparities: a community-based hepatitis-B prevention program for Asian-American communities.
Several Asian-American groups are at a higher risk of dying of liver diseases attributable to hepatitis-B infection. This culturally diverse community should be well informed of and protected against liver diseases. The present study assesses the knowledge of hepatitis B before and after a hepatitis-B educational program and determines the infection status of an Asian community. Nine Asian communities of Montgomery County, MD, enrolled in the hepatitis-B prevention program between 2005 and 2006. They attended culturally tailored lectures on prevention, completed self-administered pre- and posttests, and received blood screening for the disease. More than 800 Asian Americans participated in the study. Knowledge of prevention was improved after educational delivery. The average infection rate was 4.5%, with Cambodian, Thai, Vietnamese, Chinese and Korean groups having higher infection rates. The age group of 36-45 had the highest percentage of carriers (9.1%). Many Asian groups, particularly those of a southeast Asian decent, were subject to a higher probability of hepatitis-B infection. At an increased risk are first-generation Asian immigrants, groups with low immunization rates and those aged 36-45. The findings provide potential directions for focusing preventive interventions on at-risk Asian communities to reduce liver cancer disparities.


Student mental health implications for physician assistant education.
This report examines student mental health and learning disabilities along the continuum from childhood to college by examining diagnosing and prescribing patterns and potential implications for physician assistant (PA) faculty based on the Standards of the Accreditation Review Commission on Education for the Physician Assistant (ARC-PA) and the new Diagnostic and Statistical Manual of Mental Disorders (DSM-5, Section 2, Specific Learning Disorders). We reviewed existing data to compare decades of national patterns in diagnosing and prescribing for conditions that have known comorbidities with learning disabilities. By including quantitative and qualitative data from children, adolescents, undergraduate college students, and veterans, we illustrate the potential impact these patterns could have on some current and future applicants to PA educational programs and the requirements for PA faculty to address these needs. Multiple national data sources revealed increasing numbers of children diagnosed with Attention Deficit Hyperactivity Disorder (ADHD), psychoactive prescriptions written for those under age 18, and self-reported increases in multiple psychological disorders among college students. More recent evidence shows a twofold increase in military veterans returning to college with posttraumatic stress disorder, traumatic brain injury, and depression. The growing numbers of students with conditions described in this report requires academic faculty to be aware of requirements to recognize and address their mental health service needs. PA educators could benefit from supplemental faculty development activities to help prepare them to recognize the pipeline patterns of a growing percentage of postsecondary students with mental health conditions and learning disabilities.


Predicting Water Resource Impacts of Unconventional Gas Using Simple Analytical Equations.
The rapid expansion in unconventional gas development over the past two decades has led to concerns over the potential impacts on groundwater resources. Although numerical models are invaluable for assessing likelihood of impacts at particular sites, simpler analytical models are also useful because they help develop hydrological understanding. Analytical approaches are also valuable for preliminary assessments and to determine where more complex models are warranted. In this article, we present simple analytical solutions that can be used to predict: (1) the spatial extent of drawdown from horizontal wells drilled into the gas-bearing formation, and rate of recovery after gas production ceases; (2) the potential for upward transport of contaminants from the gas-bearing formation to shallow aquifers during hydraulic fracturing operations when pressures in the gas-bearing formation are greatly increased; and (3) the potential downward leakage of water from shallow aquifers during depressurization of gas-bearing formations. In particular, we show that the recovery of pressure after production ceases from gas-bearing shale formations may take several hundred years, and we present critical hydraulic conductivity values for intervening aquitards, below which the impact on shallow aquifers will be negligible. The simplifying assumptions inherent in these solutions will limit their predictive accuracy for site-specific assessments, compared to numerical models that incorporate knowledge of spatial variations in formation properties and which may include processes not considered in the simpler solutions.


The proteome profiles of the olfactory bulb of juvenile, adult and aged rats - an ontogenetic study.
In this study, we searched for proteins that change their expression in the olfactory bulb (oB) of rats during ontogenesis. Up to now, protein expression differences in the developing animal are not fully understood. Our investigation focused on the question whether specific proteins exist which are only expressed during different development stages. This might lead to a better characterization of the microenvironment and to a better determination of factors and candidates that influence the differentiation of neuronal progenitor cells. After analyzing the samples by two-dimensional polyacrylamide gel electrophoresis (2DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), it could be shown that the number of expressed proteins differs depending on the developmental stages. Especially members of the functional classes, like proteins of biosynthesis, regulatory proteins and structural proteins, show the highest differential expression in the stages of development analyzed. In this study, quantitative changes in the expression of proteins in the oB at different developmental stages (postnatal days (P) 7, 90 and 637) could be observed. Furthermore, the expression of many proteins was found at specific developmental stages. It was possible to identify these proteins which are involved in processes like support of cell migration and differentiation.


Coassembly Generates Peptide Hydrogel with Wound Dressing Material Properties.
Multicomponent self-assembly of peptides is a powerful strategy to fabricate novel functional materials with synergetic properties that can be used for several nanobiotechnological applications. In the present study, we used a coassembly strategy to generate an injectable ultrashort bioactive peptide hydrogel formed by mixing a dipeptide hydrogelator with a macrophage attracting short chemotactic peptide ligand. Coassembly does not impede hydrogelation as shown by cryo-transmission electron microscopy (cryo-TEM), scanning electron microscopy, and rheology. Biocompatibility was shown by cytotoxicity assays and confocal microscopy. The hydrogels release the entrapped skin antibiotic ciprofloxacin, among others, in a slow and continuous manner. Such bioinspired advanced functional materials can find applications as wound dressing materials to treat chronic wound conditions like diabetic foot ulcer.


Eye regression in blind Astyanax cavefish may facilitate the evolution of an adaptive behavior and its sensory receptors.
The forces driving the evolutionary loss or simplification of traits such as vision and pigmentation in cave animals are still debated. Three alternative hypotheses are direct selection against the trait, genetic drift, and indirect selection due to antagonistic pleiotropy. Recent work establishes that Astyanax cavefish exhibit vibration attraction behavior (VAB), a presumed behavioral adaptation to finding food in the dark not exhibited by surface fish. Genetic analysis revealed two regions in the genome with quantitative trait loci (QTL) for both VAB and eye size. These observations were interpreted as genetic evidence that selection for VAB indirectly drove eye regression through antagonistic pleiotropy and, further, that this is a general mechanism to account for regressive evolution. These conclusions are unsupported by the data; the analysis fails to establish pleiotropy and ignores the numerous other QTL that map to, and potentially interact, in the same regions. It is likely that all three forces drive evolutionary change. We will be able to distinguish among them in individual cases only when we have identified the causative alleles and characterized their effects.


Eutrophication and nutrient limitation in the aquatic zones around Huainan coal mine subsidence areas, Anhui, China.
The eutrophication of three small lakes in the aquatic zones at the Huainan coal mine subsidence areas, designated as east site (ES), central site (CS), and west site (WS), were studied. Nutrient content, species, and nitrogen (N) to phosphorus (P) ratios were obtained through water quality analyses. Nutrient limitation was evaluated by nutrient enrichment bioassays (NEBs) in the autumn of 2012 and spring of 2013. Average annual concentrations of total phosphorus (TP) were 0.05, 0.08, and 0.10 mg/L, and total nitrogen (TN) concentrations were 0.77, 1.95, and 2.06 mg/L in the water column at CS, ES, and WS, respectively. All of the three lakes exhibited 'meso-eutrophic' states and the TN:TP ratio ranged from 25:1 to 74:1 with variability between seasons and sites. NEBs verified that primary productivity in the lakes at ES and WS were mainly limited by P, while N limitation or N and P co-limitation was present in the aquatic zones at CS due to unavailable dissolved inorganic nitrogen. In the studied lakes, the blue-green algae, which comprised 70% of all identified species, was the predominant taxa, while the micro-zooplankton taxa was dominant, indicating a typical trophic structure of eutrophic lakes.


Predicting turnaround time reductions of the diagnostic track in the histopathology laboratory using mathematical modelling.
Pathology departments face a growing volume of more and more complex testing in an era where healthcare costs tend to explode and short turnaround times (TATs) are expected. In contrast, the histopathology workforce tends to shrink, so histopathology employees experience high workload during their shifts. This points to the need for efficient planning of activities in the histopathology laboratory, to ensure an equal division of workload and low TATs, at minimum costs. The histopathology laboratory of a large academic hospital in The Netherlands was analysed using mathematical modelling. Data were collected from the Laboratory Management System to determine laboratory TATs and workload performance during regular working hours. A mixed integer linear programme (MILP) was developed to model the histopathology processes and to measure the expected performance of possible interventions in terms of TATs and spread of workload. The MILP model predicted that tissue processing at specific moments during the day, combined with earlier starting shifts, can result in up to 25% decrease of TATs, and a more equally spread workload over the day. Mathematical modelling can help to optimally organise the workload in the histopathology laboratory by predicting the performance of possible interventions before actual implementation. The interventions that were predicted by the model to have the highest performance have been implemented in the histopathology laboratory of University Medical Center Utrecht. Further research should be executed to collect empirical evidence and evaluate the actual impact on TAT, quality of work and employee stress levels.


BGMUT: NCBI dbRBC database of allelic variations of genes encoding antigens of blood group systems.
Analogous to human leukocyte antigens, blood group antigens are surface markers on the erythrocyte cell membrane whose structures differ among individuals and which can be serologically identified. The Blood Group Antigen Gene Mutation Database (BGMUT) is an online repository of allelic variations in genes that determine the antigens of various human blood group systems. The database is manually curated with allelic information collated from scientific literature and from direct submissions from research laboratories. Currently, the database documents sequence variations of a total of 1251 alleles of all 40 gene loci that together are known to affect antigens of 30 human blood group systems. When available, information on the geographic or ethnic prevalence of an allele is also provided. The BGMUT website also has general information on the human blood group systems and the genes responsible for them. BGMUT is a part of the dbRBC resource of the National Center for Biotechnology Information, USA, and is available online at http://www.ncbi.nlm.nih.gov/projects/gv/rbc/xslcgi.fcgi?cmd=bgmut. The database should be of use to members of the transfusion medicine community, those interested in studies of genetic variation and related topics such as human migrations, and students as well as members of the general public.


Toward a new practical energy evaluation system for dairy cows.
Energy evaluation systems translate an animal's net energy (NE) requirements into feed metabolisable energy requirements (MER). The Feed into Milk (FiM) project (Agnew RE, Yan T, France J, Kebreab E and Thomas C 2004. Energy requirement and supply. In Feed into Milk. A new applied feeding system for dairy cows (ed. C Thomas), pp. 11-20. Nottingham University Press, Nottingham, UK) proposed a new system to predict MER of dairy cows that is, in contrast to previous energy evaluation systems for cattle, independent of feed quality. The FiM system shares this characteristic with an energy evaluation system for ad libitum-fed cattle proposed in 1994 by Tolkamp and Ketelaars (T&K). The FiM system requires nine parameters to translate requirements for NE into MER for dairy cows, while the T&K system for cattle requires only two for the same purpose. This paper analyses the contribution of each of the parameters to the final MER predictions, the differences in MER prediction between the two systems and the underlying causes of these differences. The systems differ considerably in their estimates of the NE that is required for maintenance and in their (implicit) assumptions about the partial efficiency of ME utilisation for lactation. The T&K system is based on a constant partial efficiency of ME utilisation, but in the FiM system this efficiency changes with milk yield (MY) and shows a sharp discontinuity that is at odds with the underlying biology. These are the two main causes of the differences in MER predictions. Nevertheless, over a range of MYs between 10 and 40 kg, and for cows maintaining, gaining or losing weight, the MER predictions of the two systems are very similar with maximum differences of up to ±2% only. FiM predictions of MER are systematically higher than T&K predictions for cows with very low and very high MY. It is concluded that the FiM system could reduce parameter requirements with negligible effects on MER predictions. The combination of a very high maintenance NE parameter and a curvilinear model with two subsequent corrections leads to internal inconsistencies in the FiM system. The T&K system is much simpler but it might benefit from including more recent information for the estimation of its parameters.


Human chromogranin A. Purification and characterization from catecholamine storage vesicles of human pheochromocytoma.
Chromogranin A is the quantitatively major soluble protein in catecholamine storage vesicles of the adrenal medulla and sympathetic nerve, and has been a useful index of exocytosis during sympathoadrenal neurosecretion. To probe human catecholamine storage and release, we isolated chromogranin A from chromaffin tissue in human pheochromocytoma, and compared it to chromogranin A isolated from chromaffin tissue in bovine adrenal medulla. The preparation included catecholamine storage vesicle isolation by sucrose gradient centrifugation, removal of dopamine-beta-hydroxylase by affinity chromatography on Concanavalin A-Sepharose, and preparative polyacrylamide gel electrophoresis. Human and bovine chromogranin A displayed considerable interspecies homology. Human chromogranin A is a 68,000 dalton monomeric protein with an unusual amino acid composition (31.53 weight % glutamic acid); an acidic, microheterogeneous isoelectric point (4.57-4.68); a characteristic tryptic digest peptide map; and marked dissimilarity to dopamine-beta-hydroxylase in all properties studied. A new probe of human sympathoadrenal function is available in chromogranin A.


Reversal of parkinsonian symptoms in primates by antagonism of excitatory amino acid transmission: potential mechanisms of action.
Parkinsonism is characterised by overactive glutamatergic transmission in the cortico-striatal and subthalamo-medial pallidal pathways. Local blockade of glutamatergic transmission in these pathways can alleviate parkinsonian symptoms. The effectiveness of the treatment, however, is often limited by the simultaneous appearance of unwanted side-effects. These side-effects, including ataxia and dissociative anaesthesia, are particularly problematic when N-methyl-D-aspartate (NMDA) antagonists are used. In an attempt to overcome these problems we have attempted to manipulate excitatory amino acid (EAA)-mediated neurotransmission indirectly by targeting the NMDA receptor associated modulatory sites. We review evidence which demonstrates that antagonists for both the NMDA associated glycine and polyamine sites can reverse parkinsonian symptoms when injected intra-cerebrally in both MPTP-treated and bilateral 6-OHDA lesioned marmosets without eliciting unwanted side-effects. We further review preliminary data which suggest that ifenprodil, a polyamine site antagonist, has striking anti-parkinsonian actions in the marmoset. Potential mechanisms of action underlying these effects are discussed in terms of NMDA receptor subtypes and the neuroanatomical locus of action. The anti-parkinsonian efficacy of intra-striatally administered EAA antagonists leads us to question the view of dopamine acting in the striatum as a simple neuromodulator.


Rho, Rac and Cdc42 regulate actin organization and cell adhesion in macrophages.
Rho family proteins are known to regulate actin organization in fibroblasts, but their functions in cells of haematopoietic origin have not been studied in detail. Bac1.2F5 cells are a colony-stimulating factor-1 (CSF-1)-dependent murine macrophage cell line; CSF-1 stimulates their proliferation and motility, and acts as a chemoattractant. CSF-1 rapidly induced actin reorganization in Bac1 cells: it stimulated the formation of filopodia, lamellipodia and membrane ruffles at the plasma membrane, as well as the appearance of fine actin cables within the cell interior. Microinjection of constitutively activated (V12)Rac1 stimulated lamellipodium formation and membrane ruffling. The dominant inhibitory Rac mutant, N17Rac1, inhibited CSF-1-induced lamellipodium formation, and also induced cell rounding. V12Cdc42 induced the formation of long filopodia, while the dominant inhibitory mutant N17Cdc42 prevented CSF-1-induced formation of filopodia but not lamellipodia. V14RhoA stimulated actin cable assembly and cell contraction, while the Rho inhibitor, C3 transferase, induced the loss of actin cables. Bac1 cells had cell-to-substratum adhesion sites containing beta1 integrin, pp125FAK, paxillin, vinculin, and tyrosine phosphorylated proteins. These 'focal complexes' were present in growing and CSF-1-starved cells, but were disassembled in cells injected with N17Cdc42 or N17Rac1. Interestingly, beta1 integrin did not disperse until long after focal phosphotyrosine and vinculin staining had disappeared. We conclude that in Bac1 macrophages Cdc42, Rac and Rho regulate the formation of distinct actin filament-based structures, and that Cdc42 and Rac are also required for the assembly of adhesion sites to the extracellular matrix.


Decomposition of methanol-d4 on Au-Rh bimetallic nanoclusters on a thin film of Al2O3/NiAl(100).
The decomposition of methanol-d4 that was adsorbed on Au-Rh bimetallic nanoclusters grown by the sequential deposition of Au and Rh vapors onto ordered thin-film Al2O3/NiAl(100) at 300 K, occurred by means of dehydrogenation and primarily on the surface Rh. Nevertheless, the surface Rh atoms were not equally reactive; their reactivity altered with both structural and electronic effects arising from the alloying. The Au deposited on Rh clusters decorated the surface and deactivated Rh by not only directly obstructing them but also by neighboring them. As the initially incorporated Au tended to aggregate around reactive low-coordinated Rh atoms, such as corner Rh atoms, the reactivity of the cluster, indicated by the CO and deuterium (D2) produced per surface Rh, decreased markedly. In contrast, the Rh deposited on Au clusters promoted their reactivity. The reactivity was sharply enhanced by a few incorporated Rh atoms, as they preferentially decorated the edge Au atoms, resulting in their lower coordination, more positive charge, higher energetic d-band centers, and high reactivity. On the reactive Rh, the scission of the O-D bond in the initial dehydrogenation of methanol-d4 became more preferential than the competing desorption. The further incorporated Rh failed to promote the reactivity, but the clusters remained more reactive than those formed by Rh clusters incorporating Au as their structuring involved an active atomic segregation that yielded more low-coordinated and reactive surface Rh.


Thalamocortical afferents of Lorente de Nó: medial geniculate axons that project to primary auditory cortex have collateral branches to layer I.
The injection of anterograde tracers into the ventral division of the medial geniculate body (MGV) of both rats and rabbits labels terminal axons in layer I of auditory cortex as well as the more conventional terminal arbors in layers III/IV. Whether these layer I projections represent a separate lemniscal pathway to the molecular layer or arise as collaterals of axons terminating in III/IV has not been addressed. Focal injections of the anterograde tracers biocytin or biotinylated dextran amine were made into the MGV of young rabbits. Serial section reconstruction of single MGV axons in auditory cortex revealed that layer I axons were collaterals of thalamocortical afferents that formed multiple divergent patches within III/IV. MGV collaterals to layer I often coursed tangentially for several millimeters before terminating. In some cases, the layer I collaterals descended to arborize within a thalamocortical patch in layers III/IV. These results suggest considerable radial and tangential divergence in the auditory thalamocortical pathway and argue for an expanded role for layer I in the processing of specific sensory stimuli.


Comparison of conventional susceptibility tests with direct detection of penicillin-binding protein 2a in borderline oxacillin-resistant strains of Staphylococcus aureus.
Six selected strains of Staphylococcus aureus classified as borderline oxacillin-resistant, according to standard disk diffusion and microdilution susceptibility test methods, and seven methicillin-resistant and seven methicillin-susceptible control strains were examined for the presence of penicillin-binding protein 2a (PBP 2a) by fluorography and immunoblotting and for DNA hybridization with a mec-specific probe in a dot blot assay. Oxacillin agar screen tests with and without NaCl supplementation were also performed with all strains. PBP 2a was detected both by fluorography and by immunoblotting in all seven methicillin-resistant control strains and in none of the susceptible controls. PBP 2a was detected in two borderline strains. Results of agar screen tests performed without NaCl supplementation were completely concordant with susceptibility determined by PBP 2a and mec detection methods. Agar screening with NaCl supplementation was less accurate. These findings were confirmed with 20 additional borderline strains. Direct detection methods for the presence of PBP 2a or mec, the gene encoding it, allow accurate and definitive classification of borderline strains. Further efforts to develop a rapid, clinically useful, antibody detection system for PBP 2a are warranted.


Dietary intake of antioxidant vitamins and risk of Parkinson's disease: a case-control study in Japan.
antioxidant vitamins are expected to protect cells from oxidative damage by neutralizing the effects of reactive oxygen species. However, epidemiological evidence regarding the associations between antioxidant vitamin intake and Parkinson's disease (PD) is limited and inconsistent. We investigated the relationship between dietary intake of selected antioxidant vitamins, vegetables and fruit and the risk of PD in Japan using data from a multicenter hospital-based case-control study. included were 249 patients within 6 years of onset of PD. Controls were 368 inpatients and outpatients without a neurodegenerative disease. Information on dietary factors was collected using a validated self-administered diet history questionnaire. Adjustment was made for sex, age, region of residence, pack-years of smoking, years of education, body mass index, dietary intake of cholesterol, alcohol, total dairy products, and coffee and the dietary glycemic index. higher consumption of vitamin E and β-carotene was significantly associated with a reduced risk of PD after adjustment for confounders under study: the adjusted odds ratio in the highest quartile was 0.45 (95% confidence interval [CI]: 0.25-0.79, P for trend = 0.009) for vitamin E and 0.56 (95% CI: 0.33-0.97, P for trend = 0.03) for β-carotene. Stratified by sex, such inverse associations were significant only in women. No material relationships were shown between intake of vitamin C, α-carotene, cryptoxanthin, green and yellow vegetables, other vegetables, or fruit and the risk of PD. higher intake of vitamin E and β-carotene may be associated with a decreased risk of PD.


Synthesis of thiol-containing analogues of puromycin and a study of their interaction with N-acetylphenylalanyl-transfer ribonucleic acid on ribosomes to form thioesters.
1. The thiol-containing analogue of puromycin, 6-dimethylamino-9-{1'-[3'-(2''-mercapto-3''-phenylpropionamido)-3'-deoxy-beta-d-ribofuranosyl]}purine (XVII) in which the primary amino group of the antibiotic is replaced with a thiol grouping, was synthesized chemically (compound XVII is abbreviated to thiopuromycin). 2. Thiopuromycin (XVII) was found to be active in releasing N-[(3)H]acetylphenylalanine from its tRNA carrier as the thioester, N-acetylphenylalanylthiopuromycin (XIX) in the Escherichia coli ribosomal system. The reaction product (XIX) was synthesized chemically from thiopuromycin and N-acetylphenylalanine and found to be stable to hydrolysis in the standard incubation medium at pH7.6. dl-Phenyl-lactylpuromycin (XXI), the hydroxy analogue of puromycin, was also synthesized chemically and shown to release N-acetylphenylalanine from its tRNA carrier in the E. coli ribosomal system, thus confirming the previous results of Fahnestock et al. [Biochemistry (1970) 9, 2477-2483]. 3. In marked contrast with the results obtained in the E. coli system, both thiopuromycin (XVII) and hydroxypuromycin (XXI) were found to be inactive in releasing N-acetylphenylalanine from its tRNA carrier in the rat liver ribosomal system.


GTP cyclohydrolase I expression is regulated by nitric oxide: role of cyclic AMP.
Our previous studies have demonstrated that nitric oxide (NO) leads to nitric oxide synthase (NOS) uncoupling and an increase in NOS-derived superoxide. However, the cause of this uncoupling has not been adequately resolved. The pteridine cofactor tetrahydrobiopterin (BH(4)) is a critical determinant of endothelial NOS (eNOS) activity and coupling, and GTP cyclohydrolase I (GCH1) is the rate-limiting enzyme in its generation. Thus the initial purpose of this study was to determine whether decreases in BH(4) could underlie, at least in part, the NO-mediated uncoupling of eNOS we have observed both in vitro and in vivo. Initially we evaluated the effect of inhaled NO levels on GCH1 expression and BH(4) levels in the intact lamb. Contrary to our hypothesis, we found that there was a significant increase in both plasma BH4 levels and peripheral lung GCH1 protein levels. Furthermore, in vitro, we found that exposure to the NO donor spermine NONOate (SPNONO) led to an increase in GCH1 protein and BH(4) levels in both COS-7 and pulmonary arterial endothelial cells. However, SPNONO treatment also caused a significant increase in phospho-cAMP response element binding protein (CREB) levels, as detected by Western blot analysis, and significantly increased cAMP levels, as detected by enzyme immunoassay. Furthermore, utilizing GCH1 promoter fragments fused to a luciferase reporter gene, we found that GCH1 promoter activity was enhanced by SPNONO in a CREB-dependent manner, and electromobility shift assays revealed an NO-dependent increase in the nuclear binding of CREB. These data suggest that NO increases BH(4) levels through a cAMP/CREB-mediated increase in GCH1 transcription and that the eNOS uncoupling associated with exogenous NO does not involved reduced BH(4) levels.


TWIST1 Gene: First Insights in Felis catus.
TWIST1 is thought to be a novel oncogene. Understanding the molecular mechanisms regulating the TWIST1 gene expression profiles in tumor cells may give new insights regarding prognostic factors and novel therapeutic targets in veterinary oncology. In the present study we partially isolated the TWIST1 gene in Felis catus and performed comparative studies. Several primer combinations were used based on the alignments of homologous DNA sequences. After PCR amplification, three bands were obtained, purified and sequenced. Several bioinformatic tools were utilized to carry out the comparative studies. Higher similarity was found between the isolated TWIST1 gene in Felis catus and Homo sapiens (86%) than between Homo sapiens and Rattus norvegicus or Mus musculus (75%). Partial amino acid sequence showed no change in the four species analyzed. This confirmed that coding sequences presented high similarity (~96%) between man and cat. These results give the first insights regarding the TWIST1 gene in cat but further studies are required in order to establish, or not, its role in tumor formation and progression in veterinary oncology.


First report of resistance to acetolactate-synthase-inhibiting herbicides in yellow nutsedge (Cyperus esculentus): confirmation and characterization.
Yellow nutsedge is one of the most problematic sedges in Arkansas rice, requiring the frequent use of halosulfuron (sulfonylurea) for its control. In the summer of 2012, halosulfuron at 53 g ha(-1) (labeled field rate) failed to control yellow nutsedge. The level of resistance to halosulfuron was determined in the putative resistant biotype, and its cross-resistance to other acetolactate synthase (ALS) inhibitors from four different herbicide families. ALS enzyme assays and analysis of the ALS gene were used to ascertain the resistance mechanism. None of the resistant plants was killed by halosulfuron at a dose of 13 568 g ha(-1) (256× the field dose), indicating a high level of resistance. Based on the whole-plant bioassay, the resistant biotype was not controlled by any of the ALS-inhibiting herbicides (imazamox, imazethapyr, penoxsulam, bispyribac, pyrithiobac-sodium, bensulfuron and halosulfuron) tested at the labeled field rate. The ALS enzyme from the resistant biotype was 2540 times less responsive to halosulfuron than the susceptible biotype, and a Trp574 -to-Leu substitution was detected by ALS gene sequencing using the Illumina HiSeq. The results suggest a target-site alteration as the mechanism of resistance in yellow nutsedge, which accounts for the cross-resistance to other ALS-inhibiting herbicide families.


Pervasive competition between threat and reward in the brain.
In the current functional MRI study, we investigated interactions between reward and threat processing. Visual cues at the start of each trial informed participants about the chance of winning monetary reward and/or receiving a mild aversive shock. We tested two competing hypothesis: according to the 'salience hypothesis', in the condition involving both reward and threat, enhanced activation would be observed because of increased salience; according to the 'competition hypothesis', the processing of reward and threat would trade-off against each other, leading to reduced activation. Analysis of skin conductance data during a delay phase revealed an interaction between reward and threat processing, such that the effect of reward was reduced during threat and the effect of threat was reduced during reward. Analysis of imaging data during the same task phase revealed interactions between reward and threat processing in several regions, including the midbrain/ventral tegmental area, caudate, putamen, bed nucleus of the stria terminalis, anterior insula, middle frontal gyrus and dorsal anterior cingulate cortex. Taken together, our findings reveal conditions during which reward and threat trade-off against each other across multiple sites. Such interactions are suggestive of competitive processes and may reflect the organization of opponent systems in the brain.


Minimal requirements for murine resistance to infection with Francisella tularensis LVS.
Intraperitoneal or intravenous infection of mice with Francisella tularensis LVS is lethal, with an intraperitoneal 50% lethal dose (LD50) approaching a single bacterium. Intradermal (i.d.) LVS infection has a much higher LD50, about 10(6) bacteria in BALB/cByJ mice, and survival of i.d. infection leads to solid generation of immunity against lethal challenge. To define the minimal requirements for both initial and long-term survival of i.d. infection, we characterized the nature of i.d. LVS infection in lymphocyte-deficient BALB/cByJ.scid (scid) mice. scid mice infected i.d. with strain LVS survived for about 20 days and then died from overwhelming disseminated infection. However, scid mice treated with monoclonal antibodies to gamma interferon, tumor necrosis factor alpha, or neutrophils-granulocytes all died within 1 week of infection, indicating that these were essential for early control of infection. Studies using GKO (gamma interferon knockout) mice emphasized that gamma interferon is absolutely required for initial survival of i.d. LVS infection. scid mice could be reconstituted for long-term survival of i.d. LVS infection and clearance of bacteria by intravenous transfer of splenic lymphocytes or purified B220-/T+ lymphocytes but not nu/nu lymphocytes. T cells are therefore required for long-term clearance and survival of i.d. LVS infection; efforts to determine whether CD4+ T cells, CD8+ T cells, or both are involved are ongoing.


Retention in depression treatment among ethnic and racial minority groups in the United States.
Premature discontinuation of psychiatric treatment among ethnic-racial minorities is a persistent concern. Previous research on identifying factors associated with ethnic-racial disparities in depression treatment has been limited by the scarcity of national samples with adequate representation of minority groups and especially non-English speakers. In this article, we aim to identify variations in the likelihood of retention in depression treatment among ethnic-racial minority groups in the United States as compared to non-Latino whites. Second, we aim to identify the factors that are related to treatment retention. We use data from the Collaborative Psychiatric Epidemiology Surveys to examine differences and correlates of depression treatment retention among a representative sample (n=564) of non-Latino whites, Latinos, African-American, and Asian respondents with last 12-month depressive disorder and who report receiving formal mental health treatment in the last year. We define retention as attending at least four visits or remaining in treatment during a 12-month period. Being seen by a mental health specialist as opposed to being seen by a generalist and having received medication are correlates of treatment retention for the entire sample. However, after adjusting for demographics, clinical factors including number of co-occurring psychiatric disorders and level of disability, African-Americans are significantly less likely to be retained in depression treatment as compared to non-Latino whites. Availability of specialized mental health services or comparable treatment within primary care could improve treatment retention. Low retention suggests persistent problems in the delivery of depression treatment for African-Americans.


Steric confinement and enhanced local flexibility assist knotting in simple models of protein folding.
The chaperonin complex GroEL-GroES is able to accelerate the folding process of knotted proteins considerably. However, the folding mechanism inside the chaperonin cage is elusive. Here we use a combination of lattice and off-lattice Monte Carlo simulations of simple Gō models to study the effect of physical confinement and local flexibility on the folding process of protein model systems embedding a trefoil knot in their native structure. This study predicts that steric confinement plays a specific role in the folding of knotted proteins by increasing the knotting probability for very high degrees of confinement. This effect is observed for protein MJ0366 even above the melting temperature for confinement sizes compatible with the size of the GroEL/GroES chaperonin cage. An enhanced local flexibility produces the same qualitative effects on the folding process. In particular, we observe that knotting probability increases up to 40% in the transition state of protein MJ0366 when flexibility is enhanced. This is underlined by a structural change in the transition state, which becomes devoid of helical content. No relation between the knotting mechanism and flexibility was found in the context of the off-lattice model adopted in this work.


Severe dengue categories as research endpoints-Results from a prospective observational study in hospitalised dengue patients.
Severe dengue was perceived as one clinical disease entity until the WHO 2009 classification stratified it into severe vascular leakage, severe bleeding, and severe organ dysfunction. The objectives of this study were to investigate the potential use of severe dengue categories as endpoints for intervention research. 271 patients with severe dengue among 1734 confirmed dengue patients were followed prospectively in this hospital-based observational study in Latin America and Asia. We compared the distribution of severe dengue categories according to gender and age (below/above 15y), and determined the relative frequency and the overlap of severe dengue categories in the same patients. In a next step, we extended the analysis to candidate moderate severity categories, based on recently suggested definitions which were adapted for our purposes. Severe vascular leakage occurred in 244 (90%), severe bleeding in 39 (14%), and severe organ dysfunction in 28 (10%) of 271 severe dengue patients. A higher frequency of severe leakage was seen in children or adolescents (<15y) compared to adults. More than 80% of the severe leakage cases, and 30-50% of the cases with severe bleeding or severe organ dysfunction, were defined as severe on the basis of that feature alone. In 136 out of 213 patients with severe leakage alone, neither moderate bleeding manifestation nor hepatic involvement was recorded. On the other hand, moderate leakage manifestations were detected in 4 out of 12 cases that were classified as severe based on bleeding alone. A major proportion of severe dengue patients exhibited clinical manifestations of severe vascular leakage only, which may constitute a useful endpoint for intervention research or pathophysiology studies. Severe bleeding and severe organ manifestation were recorded less frequently and exhibited a higher degree of overlap with severe leakage. Severe bleeding without leakage may be associated with individual predisposition or the presence of comorbidities. More detailed assessments are needed to explore this hypothesis. Candidate moderate disease endpoints were investigated and need to be further validated.


Structure and electronics in dimeric boron π expanded azine and salphen complexes.
Although boron-based fluorophores incorporating nitrogenous chelating ligands have received much attention, there has been little work on examples of boron-salphen and azine derivatives. In this report, we present several π expanded boron salphen type complexes, incorporating both bis(2-hydroxynaphthaldehyde)azine as well as ortho, meta and para variants of the analogous 2-hydroxynaphthaldehyde salphen compounds. For the azine, we observed only the formation of a single BF2 adduct, while for the naphth-phen compounds dimeric BF2 binding was observed. All new compounds were fully characterized via X-ray diffraction, and both DFT and TDDFT studies were carried out to probe the electronic structures of these fluorophores.


Femorotibial and patellar cartilage loss in patients prior to total knee arthroplasty, heterogeneity, and correlation with alignment of the knee.
To analyse tibial, femoral, and patellar cartilage loss in patients prior to total knee arthroplasty (TKA), and its correlation with alignment of the knee. 26 patients (aged 58 to 86 years) with a clinical indication for TKA were investigated. Quantitative end points of cartilage morphology (T scores for cartilage volume normalised to total subchondral bone area) were determined from coronal and axial magnetic resonance image data, using proprietary software. The static alignment of the knee was determined from standing full limb radiographs. The magnitude of cartilage loss (T score of normalised cartilage volume) was highly variable within the knee, correlation coefficients ranging from r = 0.17 to 0.51 between cartilage plates. The correlation of cartilage loss with static alignment of the knee (as a continuous variable) was r = -0.52 (p<0.05) for the medial tibia, -0.38 (not significant) for the medial femur, +0.76 (p<0.001) for the lateral tibia, +0.31 (not significant) for the lateral femur, and -0.09 for the patella. When analysing alignment independent of direction (valgus or varus), the correlation for the patella increased to r = 0.30, but remained non-significant. Cartilage loss was highly variable among patients and among cartilage plates before knee arthroplasty. Its correlation with alignment was stronger for the tibia than for the femur. There was some evidence for an association of alignment and patellar cartilage loss. These findings stimulate further research on the mechanism and cause-effect relation of alignment and knee osteoarthritis using quantitative magnetic resonance imaging technology.


RAS-Mediated radiation resistance is not linked to MAP kinase activation in two bladder carcinoma cell lines.
The expression of activated RAS oncogenes has been shown to increase radioresistance in a number of cell lines. The pathways by which RAS leads to radioresistance, however, are unknown. RAS activates several signal transduction pathways, with the RAF-MAP2K-MAP kinase pathway perhaps the best studied. MAP kinase has also been shown to be activated by radiation through this pathway. Given the important role of MAP kinase in multiple signaling events, we asked if radioresistance induced by RAS was mediated through the activation of MAPK. Cells of two human bladder carcinoma cell lines were used, one with a mutated oncogenic HRAS (T24) and other with a wild-type HRAS (RT4). The surviving fraction after exposure to 2 Gy of radiation (SF2) for the T24 cell lines was found to be 0.62, whereas that for RT4 cells was 0.40. Treatment with the farnesyl transferase inhibitor (FTI) L744,832, which inhibits RAS processing and activity, decreased the SF2 of T24 cells to 0.29, whereas the SF2 of RT4 cells remained unchanged after FTI treatment, thus demonstrating the importance of RAS activation to the radiosensitivity of cells with mutated RAS. MAP kinase activation was found to be constitutive and dependent on RAS in T24 cells, while it was inducible by radiation and was independent of RAS in RT4 cells. Treatment of both cell lines with the MAP2K inhibitor PD98059 inhibited MAPK activation; however, inhibiting MAPK activation had no effect on radiation survival of T24 or RT4 cells. These data indicate that MAPK activation does not contribute to RAS-induced radioresistance in this system.


Phylogenetic relationships of rollers (Coraciidae) based on complete mitochondrial genomes and fifteen nuclear genes.
The rollers (Coraciidae) constitute a relative small avian family with ca. 12 species distributed in Africa, western and southern Eurasia, and eastern Australia. In this study we examine the phylogenetic relationships of all species currently recognized in the family, including two taxa whose taxonomic status is currently contested. By using shotgun sequencing on degraded DNA from museum study skins we have been able to recover complete mitochondrial genomes as well as 15 nuclear genes for in total 16 taxa. The gene sequences were analyzed both concatenated in a maximum likelihood framework as well in a species tree approach using MP-EST. The different analytical approaches yield similar, highly supported trees and support the current division of the rollers into two genera, Coracias and Eurystomus. The only conflict relates to the placement of the Blue-bellied Roller (C. cyanogaster), where the mitochondrial, and the concatenated nuclear and mitochondrial data set, place this taxon as sister to the other Coracias species, whereas nuclear data and the species tree analysis place it as the sister taxon of C. naevia and C. spatulatus. All analyses place the Eurasian roller (C. garrulus) with the two African species, Abyssinian Roller (C. abyssinica) and Liliac-breasted Roller (C. caudatus), and place this clade as the sister group to the Asian Coracias rollers. In addition, our results support a sister group relationship between the morphologically rather dissimilar Purple Roller (C. naevia) and Racquet-tailed Roller (C. spatulatus) and also support the division of Eurystomus in an African and an Asian clade. However, within the Asian clade the Azure Roller (E. azureus) from Halmahera appears to be nested within the Dollarbird (E. orientalis), indicating that that this taxon is a morphological divergent, but a rather recent offshoot, of the widespread Dollarbird. Similarly, the Purple-winged Roller (C. temminickii) from Sulawesi group together with C. benghalensis affinis from Southeast Asia and these two in turn comprises the sister group to C. benghalensis benghalensis from India and western Asia.