Nrf2-Mediated HO-1 Induction and Antineuroinflammatory Activities of Halleridone.
Nuclear factor E2-related factor 2 (Nrf2) is the master regulator of antioxidant enzymes and is known to act on the nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) signaling pathway. Few studies have examined the bioactivity of halleridone. Herein, we investigated whether halleridone, which was isolated from the stems of the plant Cornus walteri, could regulate Nrf2-mediated heme oxygenase (HO)-1 expression and prevent intramicroglial inflammation induced by amyloid beta (Aβ)1-42 overexpression. Biochemical and molecular experiments, such as real-time polymerase chain reaction, Western blot analysis, immunocytochemistry, immunofluorescence, and luciferase reporter gene assays, were performed. The results demonstrated that halleridone promoted the upregulation of Nrf2 expression and its translocation to the nucleus, thereby activating antioxidant response element gene transcription and HO-1 expression in murine hippocampal HT22 cells. Additionally, halleridone removed intramicroglial Aβ1-42 and suppressed the production of inflammatory mediators such as interleukin (IL)-1β, IL-6, prostaglandin E2, and nitric oxide (NO) induced by artificially overexpressed Aβ1-42 and decreased pNF-κB accumulation in the nucleus and the expression of inducible NO synthase and cyclooxygenase II in BV-2 cells. In conclusion, halleridone activated Nrf2-mediated HO-1 expression and inhibited Aβ1-42-overexpressed microglial BV-2 cell activation. These observations suggest that halleridone may have therapeutic potential for targeting neurodegeneration through neuroinflammation.


N omega-nitro-L-arginine methyl ester selectively inhibits pulmonary vasodilator responses to acetylcholine and bradykinin.
The effects of N omega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of endothelium-derived relaxing factor (EDRF) production, on vascular tone and responses were investigated in the pulmonary vascular bed of the intact-chest cat under conditions of controlled blood flow and constant left atrial pressure. When pulmonary vascular tone was elevated with U-46619, intralobar injections of acetylcholine, bradykinin, sodium nitroprusside, isoproterenol, prostaglandin E1 (PGE1), lemakalim, and 8-bromo-guanosine 3',5'-cyclic monophosphate (8-bromo-cGMP) dilated the pulmonary vascular bed. Intravenous administration of L-NAME elevated lobar arterial and systemic arterial pressures without altering left atrial pressure. When U-46619 was infused after L-NAME to raise lobar arterial pressure to levels similar to those attained during the control period, vasodilator responses to acetylcholine and bradykinin were reduced significantly, whereas responses to PGE1, lemakalim, and 8-bromo-cGMP were not altered, and responses to nitroprusside were increased. There was a small effect on the response to the highest dose of isoproterenol, and pressor responses to BAY K 8644 and angiotensin II were not altered. These results are consistent with the hypothesis that EDRF production may involve the formation of nitric oxide or a nitroso compound from L-arginine and that EDRF production may have a role in the regulation of tone and in the mediation of responses to acetylcholine and bradykinin in the pulmonary vascular bed of the cat.


Principles of casting and splinting.
The ability to properly apply casts and splints is a technical skill easily mastered with practice and an understanding of basic principles. The initial approach to casting and splinting requires a thorough assessment of the injured extremity for proper diagnosis. Once the need for immobilization is ascertained, casting and splinting start with application of stockinette, followed by padding. Splinting involves subsequent application of a noncircumferential support held in place by an elastic bandage. Splints are faster and easier to apply; allow for the natural swelling that occurs during the acute inflammatory phase of an injury; are easily removed for inspection of the injury site; and are often the preferred tool for immobilization in the acute care setting. Disadvantages of splinting include lack of patient compliance and increased motion at the injury site. Casting involves circumferential application of plaster or fiberglass. As such, casts provide superior immobilization, but they are more technically difficult to apply and less forgiving during the acute inflammatory stage; they also carry a higher risk of complications. Compartment syndrome, thermal injuries, pressure sores, skin infection and dermatitis, and joint stiffness are possible complications of splinting and casting. Patient education regarding swelling, signs of vascular compromise, and recommendations for follow-up is crucial after cast or splint application.


Third-order nonlinear spectra and optical limiting of lead oxifluoroborate glasses.
We have determined two-photon absorption and nonlinear refraction spectra of the 50BO(1.5) - (50-x)PbF(2) - xPbO glasses (with x = 25, 35, 50 cationic %) at the range of the 470 and 1550 nm. The replacement of fluor atoms by oxygen leads to an increase in the third-order susceptibility, due to the formation of non-bridging oxygens (NBO). The nonlinear index of refraction is one order of magnitude higher than the one for fused silica, and it increases almost twice for the sample with x = 50. This sample has also shown promising features for all-optical switching as well as for optical limiting.


Pulotu: Database of Austronesian Supernatural Beliefs and Practices.
Scholars have debated naturalistic theories of religion for thousands of years, but only recently have scientists begun to test predictions empirically. Existing databases contain few variables on religion, and are subject to Galton's Problem because they do not sufficiently account for the non-independence of cultures or systematically differentiate the traditional states of cultures from their contemporary states. Here we present Pulotu: the first quantitative cross-cultural database purpose-built to test evolutionary hypotheses of supernatural beliefs and practices. The Pulotu database documents the remarkable diversity of the Austronesian family of cultures, which originated in Taiwan, spread west to Madagascar and east to Easter Island-a region covering over half the world's longitude. The focus of Austronesian beliefs range from localised ancestral spirits to powerful creator gods. A wide range of practices also exist, such as headhunting, elaborate tattooing, and the construction of impressive monuments. Pulotu is freely available, currently contains 116 cultures, and has 80 variables describing supernatural beliefs and practices, as well as social and physical environments. One major advantage of Pulotu is that it has separate sections on the traditional states of cultures, the post-contact history of cultures, and the contemporary states of cultures. A second major advantage is that cultures are linked to a language-based family tree, enabling the use phylogenetic methods, which can be used to address Galton's Problem by accounting for common ancestry, to infer deep prehistory, and to model patterns of trait evolution over time. We illustrate the power of phylogenetic methods by performing an ancestral state reconstruction on the Pulotu variable "headhunting", finding evidence that headhunting was practiced in proto-Austronesian culture. Quantitative cross-cultural databases explicitly linking cultures to a phylogeny have the potential to revolutionise the field of comparative religious studies in the same way that genetic databases have revolutionised the field of evolutionary biology.


Mechanism of NTP hydrolysis by the Escherichia coli primary replicative helicase DnaB protein. 2. Nucleotide and nucleic acid specificities.
The kinetic mechanism of NTP binding and hydrolysis by the Escherichia coli replicative helicase, the DnaB protein, in the absence and presence of the single-stranded DNA (ssDNA), has been quantitatively examined using the rapid quench-flow technique, under single-turnover conditions. In the case of both the free helicase and the enzyme-ssDNA complexes, the mechanism is independent of the type of base of the cofactor or the DNA; the bimolecular association is followed by the reversible chemical hydrolysis and subsequent conformational transition of the enzyme-product complex. The NTP hydrolysis step is significantly faster for the purine than for the pyrimidine cofactor, both in the absence and in the presence of the DNA. The temperature effect indicates that the nature of intermediates of the purine nucleotide, ATP, is different from the nature of the analogous intermediates of the pyrimidine nucleotide, CTP. Nevertheless, both types of cofactors seem to approach a similar "exit" state at the end of the reaction. The effect of ssDNA on the kinetics of NTP hydrolysis depends on the type of nucleotide cofactor and the base composition of the DNA and is centered at the hydrolysis step. Homoadenosine ssDNA oligomers are particularly effective in increasing the hydrolysis rate. The allosteric signal from the DNA, which activates the NTP hydrolysis, comes predominantly from the strong DNA-binding subsite. The role of the weak DNA-binding subsite is to modulate the allosteric effect of the strong subsite. The significance of these results for the mechanism of the free energy transduction by the DnaB helicase is discussed.


Statistical collaboration to impact policy decisions.
The objective of this paper is to support the following thesis: In health services research, a statistician can, and often should, contribute beyond what is generally defined as the 'classical' statistician project role. I use projects from RAND Health, the health services research division of the RAND Corporation, to demonstrate the issues that arise for the statistician in this expanded collaborative role. These projects are the 'HIV Cost and Services Utilization Study' (HCSUS) and the 'Ephedra: Clinical Efficacy and Side Effects Project'. HCSUS collected information on a nationally representative sample of people in care for HIV infection in order to determine what services were being delivered and their cost in order to guide policy decisions on the allocation of limited health care resources. The ephedra project was a systematic review conducted by the Southern California Evidence-Based Practice Centre to provide information on the existing science so that the National Institutes of Health could guide an expanded research effort to better understand the safety of ephedrine alkaloids. In order to collaborate on more than just the data analysis of a typical project, the statistician must proactively contribute to all project phases. These phases include problem formulation, study design, data collection, analysis, and the communication of results. The statistician needs to effectively and efficiently balance methodological, substantive, practical, and sometimes even ethical demands. As a result of contributing beyond standard statistical tasks, the statistician may directly impact health policy decisions.


Copper-triggered aggregation of ubiquitin.
Neurodegenerative disorders share common features comprising aggregation of misfolded proteins, failure of the ubiquitin-proteasome system, and increased levels of metal ions in the brain. Protein aggregates within affected cells often contain ubiquitin, however no report has focused on the aggregation propensity of this protein. Recently it was shown that copper, differently from zinc, nickel, aluminum, or cadmium, compromises ubiquitin stability and binds to the N-terminus with 0.1 micromolar affinity. This paper addresses the role of copper upon ubiquitin aggregation. In water, incubation with Cu(II) leads to formation of spherical particles that can progress from dimers to larger conglomerates. These spherical oligomers are SDS-resistant and are destroyed upon Cu(II) chelation or reduction to Cu(I). In water/trifluoroethanol (80:20, v/v), a mimic of the local decrease in dielectric constant experienced in proximity to a membrane surface, ubiquitin incubation with Cu(II) causes time-dependent changes in circular dichroism and Fourier-transform infrared spectra, indicative of increasing beta-sheet content. Analysis by atomic force and transmission electron microscopy reveals, in the given order, formation of spherical particles consistent with the size of early oligomers detected by gel electrophoresis, clustering of these particles in straight and curved chains, formation of ring structures, growth of trigonal branches from the rings, coalescence of the trigonal branched structures in a network. Notably, none of these ubiquitin aggregates was positive to tests for amyloid and Cu(II) chelation or reduction produced aggregate disassembly. The early formed Cu(II)-stabilized spherical oligomers, when reconstituted in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes and in POPC planar bilayers, form annular and pore-like structures, respectively, which are common to several neurodegenerative disorders including Parkinson's, Alzheimer's, amyotrophic lateral sclerosis, and prion diseases, and have been proposed to be the primary toxic species. Susceptibility to aggregation of ubiquitin, as it emerges from the present study, may represent a potential risk factor for disease onset or progression while cells attempt to tag and process toxic substrates.


The mechanism of reducing thiopentone dose in elderly patients undergoing surgery.
The dose of thiopentone required to induce anesthesia in adults decreasing with age is not due to pharmacodynamic change. The change of pharmacokinetic properties of thiopentone with age in undergoing surgery patient's arterial blood was investigated in seven elderly (67-82 yr) and six young (21-33 yr) patients of both sexes. Thiopentone (3 mg kg-1) was administered intravenously and arterial blood samples were obtained immediately after the injection to measure plasma and red blood cell thiopentone concentrations by an HPLC method. Plasma protein binding was studied using ultracentrifuge method. The disappearance of thiopentone from the arterial blood was described by a two-compartment open model. The distribution rate constant (alpha) was significantly larger in the young patients (p less than 0.001). The distribution half-life was longer in the elderly (p less than 0.05). Both the input microscopic rate constant, K21, and the exit microscopic rate constant, K12, with the central compartment were significantly larger in the young patients. (p less than 0.02 and p less than 0.001, respectively). The difference between the exit and input microscopic rate constant, K12-K21, was much larger in the young patients (p less than 0.001). The plasma protein binding was significantly reduced in the elderly (p less than 0.05). The apparent overall volume of distribution, Vd was not significantly different between young and elder patients. However, the volume of distribution of the central compartment was smaller in the young patients (p less than 0.05). This was probably due to the difficulty of estimation of initial thiopentone plasma concentration post-equilibrium in the central compartment after administration of thiopentone.(ABSTRACT TRUNCATED AT 250 WORDS)


Longitudinal Changes in Parkinson's Disease Symptoms with and Without Rapid Eye Movement Sleep Behavior Disorder: The Oxford Discovery Cohort Study.
Parkinson's disease (PD) comorbid with rapid eye movement sleep behavior disorder (RBD) may show more severe motor and nonmotor symptoms, suggesting a distinct PD subtype. The aim of this study was to investigate the impact of RBD on the longitudinal change of motor and nonmotor symptoms in patients with PD. Patients with early PD (diagnosed within 3.5 years) recruited from 2010 to 2019 were followed every 18 months in the Oxford Parkinson's Disease Centre Discovery cohort. At each visit, we used standard questionnaires and measurements to assess demographic features and motor and nonmotor symptoms (including RBD, daytime sleepiness, mood, autonomic symptoms, cognition, and olfaction). Data were analyzed with linear mixed effects and Cox regression models. Possible RBD (pRBD) was longitudinally determined according to RBD Screening Questionnaire scores. A total of 923 patients were recruited (mean age: 67.1 ± 9.59 years; 35.9% female), and 788 had follow-up assessment(s) (mean: 4.8 ± 1.98 years, range: 1.3-8.3). Among them, 33.3% were identified as pRBD (PD + pRBD). Patients with PD + pRBD had more severe baseline symptoms and showed faster progression on Movement Disorder Society-Unified Parkinson's Disease Rating Scale parts I and III, Purdue Pegboard test, and Beck Depression Inventory scores. Moreover, PD + pRBD was associated with an increased level of risk for mild cognitive impairment (hazard ratio [HR] = 1.36, 95% confidence interval [CI]: 1.01-1.83), freezing of gait (HR = 1.42, 95% CI: 1.10-1.86), and frequent falling (HR = 1.62, 95% CI: 1.02-2.60). Patients with PD + pRBD progress faster on motor, mood, and cognitive symptoms, confirming a more aggressive PD subtype that can be identified at baseline and has major clinical implications. © 2021 International Parkinson and Movement Disorder Society.


A difference between the inheritance of classical juvenile-onset and maturity-onset type diabetes of young people.
A difference in the inheritance of diabetes has been shown between the families of twenty-six patients with maturity-onset type diabetes of young people (MODY) and families of thirty-five patients with classical juvenile-onset diabetes (JOD). In the families of MODY: 1) twenty-two of twenty-six (85 per cent) propositi had a diabetic parent; 2) 46 per cent of families showed direct vertical transmission of diabetes through three generations; 3) of forty-seven tested siblings twenty-five (53 per cent) had latent diabetes; 4) the diabetic phenotype in the families was consistent, most affected individuals having a noninsulin requiring type of disease. These findings are compatible with autosomal dominant inheritance of MODY, although they do not exclude multifactorial inheritance. In contrast, in the families of JOD: 1) only four (11 per cent) of propositi had a diabetic parent; 2) three generation inheritance was found in only two (6 per cent) of JOD families, and 3) of seventy-four tested siblings eight (11 per cent) were diabetic. This difference provides further evidence of genetic heterogeneity in diabetes mellitus and indicates that there is a need for careful definition of the phenotype of diabetes in populations in which the genetics of diabetes is to be analyzed. Diabetes 24:44-53, January, 1975.


Nucleotide insertion kinetics opposite abasic lesions in DNA.
A gel assay is introduced to measure DNA polymerase insertion kinetics at single sites along a DNA template strand. The assay is used to analyze the kinetics of inserting deoxynucleotides opposite a synthetic abasic (apurinic/apyrimidinic) lesions using Drosophila DNA polymerase alpha. The location of the abasic lesion next to different nearest-neighbor bases allows the effects of base stacking on the specificity of insertion to be evaluated. The specificity of nucleotide insertion, Vmax/Km, is 6-11 times greater for A over G and about 20-50 times greater for A over C and T. The insertion specificity at the abasic lesion appears to depend more on differences in Vmax than Km. Apparent Michaelis constants for inserting A and G deoxynucleotides are similar to within about a factor of 2. The insertion of A or G occurs most efficiently at the abasic lesion when T is the 5'-nearest neighbor on the primer strand and least efficiently when G is the 5'-nearest neighbor. The presence of different base stacking partners adjacent to the site of insertion has up to a 4-fold effect on specificity.


Explosive transitions induced by interdependent contagion-consensus dynamics in multiplex networks.
We introduce a model to study the interplay between information spreading and opinion formation in social systems. Our framework consists in a two-layer multiplex network where opinion dynamics takes place in one layer, while information spreads on the other one. The two dynamical processes are mutually coupled in such a way that the control parameters governing the dynamics of the node states at one layer depend on the dynamical states at the other layer. In particular, we consider the case in which consensus is favored by the common adoption of information, while information spreading is boosted between agents sharing similar opinions. Numerical simulations of the model point out that, when the coupling between the dynamics of the two layers is strong enough, a double explosive transition, i.e., a discontinuous transition both in consensus dynamics and in information spreading appears. Such explosive transitions lead to bi-stability regions in which the consensus-informed states and the disagreement-uninformed states are both stable solutions of the intertwined dynamics.


Latest approaches to treating atopic dermatitis.
Recent investigations have shown atopic dermatitis (AD) results from the interplay of epidermal barrier defects, immune dysfunction and environmental triggers. These discoveries teach the importance of addressing all of these factors in treating patients. The barrier issues remind us of the need for moisturizers, especially after bathing, to replenish lipids and decrease water loss. Addition of dilute sodium hypochlorite (bleach) to bathwater often improves the dermatitis of children with moderate-to-severe dermatitis, especially with a history of staphylococcal infection. Oral vitamin D(3) has been suggested to increase the deficient antimicrobial peptide that results from cutaneous inflammation, although studies to date are inconclusive. Topical corticosteroids and calcineurin inhibitors continue to predominate as therapy and, to date, their safety record seems good, despite the Black Box warning issued in the United States. Poor compliance may explain the failure to respond to therapy, and contact dermatitis from topical application of emollients and medications may mimic AD, erroneously suggesting recalcitrance. For patients with severe AD, administration of systemic immunosuppressants may be required; ongoing studies of newer agents, including biologics, may revolutionize therapy for individuals with severe dermatitis, providing more targeted therapy in the future.


Brain metastasis-related microRNAs in patients with advanced breast cancer.
Brain metastasis is a major distant metastasis occurring in patients with advanced breast cancer, and is associated with poor prognosis. MicroRNAs (miRNAs) have a strong influence on various oncological functions and have been reported as potential biomarkers for detecting distant metastasis. Specific biomarkers and unique miRNAs for brain metastasis have yet to be reported. The aim of this study was to identify novel miRNAs in serum, to assist in the diagnosis of brain metastasis in patients with advanced breast cancer. We retrospectively analyzed the medical records of patients with breast cancer and collected clinical data. In addition, we evaluated serum miRNA profiles in patients with breast cancer, with and without brain metastasis, using high-sensitivity microarrays. All patients underwent computed tomography or magnetic resonance imaging brain imaging tests. A total of 51 serum samples from patients with breast cancer and brain metastasis, stored in the National Cancer Center Biobank, were used, and 28 serum samples were obtained from controls without brain metastasis. Two miRNAs, miR-4428 and miR-4480, could significantly distinguish patients with brain metastasis, with area under the receiver operating characteristic curve (AUC) values of 0.779 and 0.781, respectively, while a combination of miR-4428 and progesterone receptor had an AUC value of 0.884. No significant correlations were identified between the expression levels of these two miRNAs in serum and clinical data. We conclude that serum miR-4428 and miR-4480 may be useful as biomarkers for predicting brain metastasis in patients with breast cancer.


Differences in outcomes between older community-dwelling patients with bipolar disorder and schizophrenia with illness onset at young age.
Older community-dwelling patients with severe mental illness (SMI), particularly those with illness onset at young age, constitute a group of survivors with unique long-term care needs. Using an Asian sample in Taiwan, we attempted to find out the differences in outcomes related to physical health, cognition, and social functioning between older community-dwelling adults with bipolar disorder and schizophrenia with early age onset. Community-dwelling patients aged >50 years with bipolar I disorder or schizophrenia whose illness developed before the age of 40 years were recruited. Clinical data were obtained by reviewing all available medical records and by interviewing the patients and their reliable family members. Medical morbidities, Mini-Mental State Examination (MMSE), Cumulative Illness Rating Scale for Geriatrics (CIRS-G), and Global Assessment of Functioning (GAF) scores were compared between the two groups. In total, 113 bipolar patients and 104 schizophrenic ones (mean ages = 59.8 and 59.2 years, respectively) became the final subjects. The rates of cognitive impairment (MMSE score <24) were comparable in bipolar disorder (26.5%) and schizophrenia (24.0%) and the mean MMSE scores did not significantly differ from each other. The concurrence (54.9%) of cardiovascular disease (CVD) in the bipolar group was also similar to 51.0% in the schizophrenic one. In a multiple logistic regression analysis, the bipolar group exhibited significantly higher CIRS-G total scores (95% confidence interval (CI) for odds ratio (OR) = 1.01-1.27), body mass index (95% CI for OR = 1.02-1.21), and GAF scores (95% CI for OR = 1.04-1.14). Given better social functioning and the same cognitive function in older community-dwelling patients with bipolar disorder, they may remain at higher risk for obesity and medical morbidity than schizophrenic patients. Treatments targeting cognitive impairment and CVDs across their life span are both necessary to promote the health of community-dwellers with SMI.


Tissue regeneration in bone defects adjacent to endosseous implants: an experimental pilot study.
Extraction socket defects or alveolar ridge defects may limit or restrict placement of implants. The present study assessed the efficacy of inducing bone formation with demineralized freeze-dried bone (DFDB), osteoinduction with hydroxyapatite (HA), and guided bone regeneration with expanded polytetrafluoroethylene (e-PTFE) in bone defects adjacent to endosseous implants in an animal model. Twenty-four implants were placed in the long bones of 3 pigs. After preparation of 24 cylindric acute defects adjacent to the coronal 5 mm of each implant, 18 of the defects were treated by DFDB, HA, or e-PTFE. The 6 remaining defects healed spontaneously with no treatment (controls). In each pig, one tibia received 4 commercially pure titanium screw implants (3.75 mm x 15 mm), while the contralateral tibia received 4 HA-coated cylindric implants (3.25 mm x 15 mm). Bone formation in response to the 3 treatments and control cases was evaluated using computer-assisted densitometric image analysis. The results showed that a higher density of tissue formed under e-PTFE membranes. A significantly greater increase in bone density was noticed in bone defects treated with an e-PTFE membrane vs DFDB (P = 0.04), in e-PTFE vs control sites (P = 0.04), and in defects treated with HA vs DFDB (P = 0.04) in HA-coated implants. In titanium implants the differences observed were not significant. It was also observed that bone regeneration in defects treated with HA differs with the type of implant used (titanium vs HA-coated implants), but such differences were not significant.


Biotinylation in vivo as a sensitive indicator of protein secretion and membrane protein insertion.
Escherichia coli biotin ligase is a cytoplasmic protein which specifically biotinylates the biotin-accepting domains from a variety of organisms. This in vivo biotinylation can be used as a sensitive signal to study protein secretion and membrane protein insertion. When the biotin-accepting domain from the 1.3S subunit of Propionibacterium shermanii transcarboxylase (PSBT) is translationally fused to the periplasmic proteins alkaline phosphatase and maltose-binding protein, there is little or no biotinylation of PSBT in wild-type E. coli. Inhibition of SecA with sodium azide and mutations in SecB, SecD, and SecF, all of which slow down protein secretion, result in biotinylation of PSBT. When PSBT is fused to the E. coli inner membrane protein MalF, it acts as a topological marker: fusions to cytoplasmic domains of MalF are biotinylated, and fusions to periplasmic domains are generally not biotinylated. If SecA is inhibited by sodium azide or if the SecE in the cell is depleted, then the insertion of the MalF second periplasmic domain is slowed down enough that PSBT fusions in this part of the protein become biotinylated. Compared with other protein fusions that have been used to study protein translocation, PSBT fusions have the advantage that they can be used to study the rate of the insertion process.


Retinal laser photocoagulation. Effect of rate of energy delivery.
Three monkeys were treated with krypton red laser photocoagulation in a manner simulating treatment of a choroidal neovascular membrane. One eye of each monkey was treated by employing immediately the power level needed to produce a lesion judged biomicroscopically to be of sufficient intensity (Method A). The fellow eye was treated by starting with lower power levels and then gradually increasing to the level needed to produce a similar-appearing lesion (Method B). Parameters of spot size and treatment duration were kept constant. Animals were sacrificed three days, eight days, and four months after treatment. Both methods of treatments spared the overlying ganglion cell layer in all fundus locations, but Method A induced cell loss in the inner nuclear layer while Method B spared it. Since reaching the clinical endpoint for Method B required fewer spots at the highest power level, the number of spots at the highest power level is probably the most important factor in determining degree of inner retinal absorption. Since both methods of treatment resulted in satisfactory closure in the choriocapillaries, the histopathologic differences may not be clinically important.


Synthesis of magnetic ion-imprinted composites and selective separation and preconcentration of U(VI).
The U(VI) magnetic ion-imprinted composite (MIIC) with a uniform core-shell structure for the selective separation and preconcentration of U(VI) was prepared by copolymerization of a ternary complex of uranyl ions with 4-vinylpyridine (4-VP) and acrylamide in the presence of 2,2'-azobisisobutyronitrile. The sorption of U(VI) on the MIIC from aqueous solution was evaluated. The maximum sorption capacity of MIIC for U(VI) was 354.85 mg g(-1), which was much higher than that of the magnetic nonimprinted composite. The MIIC could be recovered by desorbing the U(VI)-loaded MIIC with 0.5 mol L(-1) HNO3, and the surface morphology of MIIC after five consecutive sorption/desorption cycles was significantly damaged. The competitive sorption experiments showed that the MIIC had a desirable selectivity for U(VI) over a range of competing metal ions. The MIIC may be a promising sorbent material for the selective separation and preconcentration of U(VI).


Mesoscale characterization of supramolecular transient networks using SAXS and rheology.
Hydrogels and, in particular, supramolecular hydrogels show promising properties for application in regenerative medicine because of their ability to adapt to the natural environment these materials are brought into. However, only few studies focus on the structure-property relationships in supramolecular hydrogels. Here, we study in detail both the structure and the mechanical properties of such a network, composed of poly(ethylene glycol), end-functionalized with ureido-pyrimidinone fourfold hydrogen bonding units. This network is responsive to triggers such as concentration, temperature and pH. To obtain more insight into the sol-gel transition of the system, both rheology and small-angle X-ray scattering (SAXS) are used. We show that the sol-gel transitions based on these three triggers, as measured by rheology, coincide with the appearance of a structural feature in SAXS. We attribute this feature to the presence of hydrophobic domains where cross-links are formed. These results provide more insight into the mechanism of network formation in these materials, which can be exploited for tailoring their behavior for biomedical applications, where one of the triggers discussed might be used.


Inhibitory self-control moderates the effect of changed implicit food evaluations on snack food consumption.
The current study used a modified implicit association test (IAT) to change implicit evaluations of unhealthy snack food and tested its effects on subsequent consumption. Furthermore, we investigated whether these effects were moderated by inhibitory self-control. A sample of 148 women (17-25 years) motivated to manage weight through healthy eating completed an IAT intervention, and pre- and post-intervention IATs assessing implicit evaluations of unhealthy food. The intervention IAT trained participants to pair unhealthy food stimuli with either positive or negative stimuli. A task disguised as a taste-test was used to assess consumption of unhealthy snack foods. Inhibitory self-control was measured using a self-report scale. As predicted, the implicit evaluation of unhealthy food became more negative from pre- to post-training among participants in the food negative pairing condition; however, there was no corresponding change in the food positive pairing condition. The effect of the training on snack consumption was moderated by inhibitory self-control with only participants low in inhibitory self-control having lower snack intake following the food negative training. This finding is consistent with dual-process models of behaviour which predict that self-control capacity renders impulses less influential on behaviour. Furthermore, it suggests that an intervention that retrains implicit food evaluations could be effective at reducing unhealthy eating, particularly among those with low inhibitory self-control.


Overlapping photoprotective function of vitamin E and carotenoids in Chlamydomonas.
Tocopherols (vitamin E) and carotenoids are the two most abundant groups of lipid-soluble antioxidants in the chloroplast. Carotenoids are well known for their roles in protecting against photooxidative stress, whereas the photoprotective functions of tocopherols have only recently been examined experimentally. In addition, little is known about the functional overlap of carotenoids and tocopherols in vivo. To investigate this possible overlap, Chlamydomonas reinhardtii strains were engineered to overproduce tocopherols by chloroplast transformation with non-codon-optimized and codon-optimized versions of the homogentisate phytyltransferase vitamin E2 (VTE2) from Synechocystis and by nuclear transformation with VTE2 from C. reinhardtii, which resulted in 1.6-fold, 5-fold to 10-fold, and more than 10-fold increases in total tocopherol content, respectively. To test if tocopherol overproduction can compensate for carotenoid deficiency in terms of antioxidant function, the nuclear VTE2 gene from C. reinhardtii was overexpressed in the npq1 lor1 double mutant, which lacks zeaxanthin and lutein. Following transfer to high light, the npq1 lor1 strains that overaccumulated tocopherols showed increased resistance for up to 2 d and higher efficiency of photosystem II, and they were also much more resistant to other oxidative stresses. These results suggest an overlapping functions of tocopherols and carotenoids in protection against photooxidative stress.


[Validation of a method for monitoring of genetically engineered human insulin manufacture].
A method for monitoring the manufacture of genetically engineered human insulin by HPLC was developed. The method was validated by the estimation of its linearity, correctness, accuracy, specificity, and stability; the limits of detection and quantitative assessment were also determined. It was proven that HPLC analysis enables reliable and reproducible results to be obtained and can be used for monitoring insulin manufacture.


Attachment and hikikomori: a psychosocial developmental model.
Hikikomori (acute social withdrawal) is a social issue in Japan that affects both the society and the lives of the individual sufferers. This study aims to connect attachment theory and hikikomori by using a culturally sensitive psychosocial developmental model that outlines the various stages of attachment throughout the developmental years. Twenty-four hikikomori sufferers and 60 comparison group participants were given questionnaires assessing parent and peer relationships, temperament and school experiences. We found the hikikomori participants had a higher incidence of ambivalent attachment, reported more parental and peer rejection and bullying, and expressed greater temperamental shyness. Path analysis supported our developmental model. We found that shy temperament and parental rejection predicted ambivalent attachment, which when coupled with peer rejection predicted hikikomori. Our model implies that treatment and prevention may require attention to attachment insecurities in early childhood, peer rejection in middle childhood and/or early adolescence. We believe it is helpful in understanding hikikomori to first understand how the attachment system balances security with exploration and the anxiety associated with novelty and challenge. Finally, we examine implications of the model for intervention, treatment and future research.


Validation of the Tracmor triaxial accelerometer system for walking.
Walking is likely to contribute substantially to nonexercise activity thermogenesis. The Tracmor triaxial accelerometer system (Maastricht, The Netherlands) is the most widely validated system for detecting body movement in free-living subjects. The aim of this study was to validate the Tracmor triaxial accelerometer system for estimating the energy expenditure of walking. Experiments were conducted in healthy subjects. First, baseline variability for Tracmor output was determined for subjects standing still. Second, Tracmor output was compared for walking on a treadmill and on level ground. Third, both Tracmor output and energy expenditure were compared for walking on a treadmill and walking on level ground. Finally, the effect of gradient on Tracmor output and energy expenditure was compared for subjects walking on a treadmill. The data demonstrated excellent reproducibility for comparing Tracmor output for standing (CV < 2%). There were excellent log-linear relationships between velocity and Tracmor output walking on a treadmill (r = 0.998) and on level ground (r = 0.999). Tracmor output and the energy expenditure of walking were inseparable for the two modalities of walking. However, the variance in response was such that to reliably derive the relationship between Tracmor output and energy expenditure, separate regression equations are needed for each subject. Finally, the Tracmor accelerometer did not detect the increased energy expenditure of walking that occurs as gradient increases. The Tracmor triaxial accelerometer provides reproducible and reliable data on the body motion associated with walking regardless of whether a subject walks on a treadmill or level ground. Tracmor units can be used to predict the energetic cost of walking provided that separate regression equations are derived for each subject to convert Tracmor output to energy expenditure.


Multipolar effects on the dipolar polarizability of magneto-electric antennas.
We show the important role played by the multipolar coupling between the illuminating field and magneto-electric scatterers even in the small particle limit (λ/10). A general multipolar method is presented which, for the case of planar non centrosymmetric particles, generates a simple expression for the polarizability tensor that directly links the dipolar moment to the incident field. The relevancy of this approach is demonstrated by comparing thoroughly the dipolar moments predicted by the method with full numerical calculations.


Experimental and numerical analysis on temporal dynamics of polarization switching in an injection-locked 1.55-μm wavelength VCSEL.
We present experimental and numerical study of temporal characteristics of injection-locked polarization switching of a conventional-type 1.55-μm wavelength single-mode vertical cavity surface-emitting laser (VCSEL). Delayed recovery response of the main-mode of the VCSEL was observed when short and strong optical injection pulses of an orthogonal polarization to the VCSEL's main polarization-mode were applied. Numerical analysis based on a spin-flip model describes that the relatively long upper level lifetimes compared to a short injection-pulse width and long cavity photon lifetimes cause delayed recovery response of the main-mode of the VCSEL. An optimum bias current of the VCSEL was also observed for the shortest recovery time of its free-running polarization mode after the orthogonal polarization beam pulse injection.


Increased cell-to-cell variation in gene expression in ageing mouse heart.
The accumulation of somatic DNA damage has been implicated as a cause of ageing in metazoa. One possible mechanism by which increased DNA damage could lead to cellular degeneration and death is by stochastic deregulation of gene expression. Here we directly test for increased transcriptional noise in aged tissue by dissociating single cardiomyocytes from fresh heart samples of both young and old mice, followed by global mRNA amplification and quantification of mRNA levels in a panel of housekeeping and heart-specific genes. Although gene expression levels already varied among cardiomyocytes from young heart, this heterogeneity was significantly elevated at old age. We had demonstrated previously an increased load of genome rearrangements and other mutations in the heart of aged mice. To confirm that increased stochasticity of gene expression could be a result of increased genome damage, we treated mouse embryonic fibroblasts in culture with hydrogen peroxide. Such treatment resulted in a significant increase in cell-to-cell variation in gene expression, which was found to parallel the induction and persistence of genome rearrangement mutations at a lacZ reporter locus. These results underscore the stochastic nature of the ageing process, and could provide a mechanism for age-related cellular degeneration and death in tissues of multicellular organisms.


Meningococcal Outer Membrane Vesicle Composition-Dependent Activation of the Innate Immune Response.
Meningococcal outer membrane vesicles (OMVs) have been extensively investigated and successfully implemented as vaccines. They contain pathogen-associated molecular patterns, including lipopolysaccharide (LPS), capable of triggering innate immunity. However, Neisseria meningitidis contains an extremely potent hexa-acylated LPS, leading to adverse effects when its OMVs are applied as vaccines. To create safe OMV vaccines, detergent treatment is generally used to reduce the LPS content. While effective, this method also leads to loss of protective antigens such as lipoproteins. Alternatively, genetic modification of LPS can reduce its toxicity. In the present study, we have compared the effects of standard OMV isolation methods using detergent or EDTA with those of genetic modifications of LPS to yield a penta-acylated lipid A (lpxL1 and pagL) on the in vitro induction of innate immune responses. The use of detergent decreased both Toll-like receptor 4 (TLR4) and TLR2 activation by OMVs, while the LPS modifications reduced only TLR4 activation. Mutational removal of PorB or lipoprotein factor H binding protein (fHbp), two proteins known to trigger TLR2 signaling, had no effect, indicating that multiple TLR2 ligands are removed by detergent treatment. Detergent-treated OMVs and lpxL1 OMVs showed similar reductions of cytokine profiles in the human monocytic cell line MM6 and human dendritic cells (DCs). OMVs with the alternative penta-acylated LPS structure obtained after PagL-mediated deacylation showed reduced induction of proinflammatory cytokines interleukin-6 (IL-6) and IL-1β but not of IP-10, a typical TRIF-dependent chemokine. Taken together, these data show that lipid A modification can be used to obtain OMVs with reduced activation of innate immunity, similar to what is found after detergent treatment.


MURC, a muscle-restricted coiled-coil protein that modulates the Rho/ROCK pathway, induces cardiac dysfunction and conduction disturbance.
We identified a novel muscle-restricted putative coiled-coil protein, MURC, which is evolutionarily conserved from frog to human. MURC was localized to the cytoplasm with accumulation in the Z-line of the sarcomere in the murine adult heart. MURC mRNA expression in the heart increased during the developmental process from the embryonic stage to adulthood. In response to pressure overload, MURC mRNA expression increased in the hypertrophied heart. Using the yeast two-hybrid system, we identified the serum deprivation response (SDPR) protein, a phosphatidylserine-binding protein, as a MURC-binding protein. MURC induced activation of the RhoA/ROCK pathway, which modulated serum response factor-mediated atrial natriuretic peptide (ANP) expression and myofibrillar organization. SDPR augmented MURC-induced transactivation of the ANP promoter in cardiomyocytes, and RNA interference of SDPR attenuated the action of MURC on the ANP promoter. Transgenic mice expressing cardiac-specific MURC (Tg-MURC) exhibited cardiac contractile dysfunction and atrioventricular (AV) conduction disturbances with atrial chamber enlargement, reduced thickness of the ventricular wall, and interstitial fibrosis. Spontaneous episodes of atrial fibrillation and AV block were observed in Tg-MURC mice. These findings indicate that MURC modulates RhoA signaling and that MURC plays an important role in the development of cardiac dysfunction and conduction disturbance with increased vulnerability to atrial arrhythmias.


Radiographic indices in one hundred fifty normal Iranian wrists.
Radiography is the most widely available imaging modality. Precise evaluations of wrist x-ray can help diagnosis and evaluate the prognosis of many wrist disorders. We measured length, angles and indices in 150 posteroanterior and lateral wrist x-rays to determine normal dimensions and variations according to age and sex. All x-rays were made with standard exposure, with the wrist and forearm in a neutral position. The average carpal height ratio was 0.52±0.03 with the Youm method and 1.5±0.09 with the Nattrass method. Mean ulnar variance was +0.99±1.6 mm and mean radial inclination was 25±4 degrees. The average radial tilt was 10±5.1 degrees. Mean scapholunate angle was 50±8.4 degrees (normal range 40-60). Carpal height, third metacarpal and capitate length were smaller in women than in men. There was a significant positive relationship between all dimensions. Our data base may be used to follow-up in conditions such as carpal instability, osteoarthritis and osteonecrosis, as well as for clinical research.


Cyberbullying and adolescent mental health.
The rapid growth of electronic and computer-based communication and information sharing during the past decade has dramatically changed social interactions, especially among teenagers. Cyberbullying has emerged as a new form of bullying and harassment, and it has been shown to possess different ramifications from traditional school-yard bullying. This problem has emerged in nations worldwide. Cyber victims have reported various emotional and behavioral symptoms, along with school-related problems. This paper reviews international cross-sectional studies relating to the definition, prevalence, age, and gender differences inherent in cyberbullying. Psychosocial and risk factors associated with cyberbullying are also addressed. Prevention and intervention strategies for school officials and parents are suggested. Healthcare providers, policy makers, and families must be ever-mindful of the grave dangers cyberbullying poses to youths. Longitudinal studies are warranted to assess the psychological risk factors of cyberbullying.


Access to HIV/AIDS care: a systematic review of socio-cultural determinants in low and high income countries.
The role of socio-cultural factors in influencing access to HIV/AIDS treatment, care and support is increasingly recognized by researchers, international donors and policy makers. Although many of them have been identified through qualitative studies, the evidence gathered by quantitative studies has not been systematically analysed. To fill this knowledge gap, we did a systematic review of quantitative studies comparing surveys done in high and low income countries to assess the extent to which socio-cultural determinants of access, identified through qualitative studies, have been addressed in epidemiological survey studies. Ten electronic databases were searched (Cinahl, EMBASE, ISI Web of Science, IBSS, JSTOR, MedLine, Psyinfo, Psyindex and Cochrane). Two independent reviewers selected eligible publications based on the inclusion/exclusion criteria. Meta-analysis was used to synthesize data comparing studies between low and high income countries. Thirty-four studies were included in the final review, 21 (62%) done in high income countries and 13 (38%) in low income countries. In low income settings, epidemiological research on access to HIV/AIDS services focused on socio-economic and health system factors while in high income countries the focus was on medical and psychosocial factors. These differences depict the perceived different barriers in the two regions. Common factors between the two regions were also found to affect HIV testing, including stigma, high risk sexual behaviours such as multiple sexual partners and not using condoms, and alcohol abuse. On the other hand, having experienced previous illness or other health conditions and good family communication was associated with adherence to ART uptake. Due to insufficient consistent data, a meta-analysis was only possible on adherence to treatment. This review offers evidence of the current challenges for interdisciplinary work in epidemiology and public health. Quantitative studies did not systematically address in their surveys important factors identified in qualitative studies as playing a critical role on the access to HIV/AIDS services. The evidences suggest that the problem lies in the exclusion of the qualitative information during the questionnaire design. With the changing face of the epidemic, we need a new and improved research strategy that integrates the results of qualitative studies into quantitative surveys.


A cost-effectiveness analysis of teledietetics in short-, intermediate-, and long-term weight reduction.
To evaluate the effectiveness of teledietetics in weight loss for 24 weeks and the cost-effectiveness of weight loss between face-to-face and teledietetics services. The study was conducted at a community health center and a community dietetics clinic. The study was a quasiexperimental design. Fifty adults aged 20-50 with a BMI ≥23 participated in the study. The face-to-face (FD) group received 12 dietary counselling sessions and recorded their diet in a log book. The teledietetics (TD) group attended three group nutrition seminars and recorded their diet on a Web-based platform. Changes in variables were compared using an independent t-test. Direct and indirect costs were applied to compute cost-effectiveness ratios. At week 6, the FD group showed greater reductions in all variables than did the TD group. At week 12, the effects reversed. At week 24, the accumulated reductions in weight and fat in the TD group were significantly higher than those in the FD group (all at p < 0.0001). The observed direct costs for 1% weight loss and 1% fat loss were USD 28.24 and USD17.09, respectively. A dietetic service delivered as a teledietetics model is more cost-effective than the face-to-face dietetics model in weight reduction.


Impact of dizziness on everyday life in older primary care patients: a cross-sectional study.
Dizziness is a common and often disabling symptom, but diagnosis often remains unclear; especially in older persons where dizziness tends to be multicausal. Research on dizziness-related impairment might provide options for a functional oriented approach, with less focus on finding diagnoses. We therefore studied dizziness-related impairment in older primary care patients and aimed to identify indicators related to this impairment. In a cross-sectional study we included 417 consecutive patients of 65 years and older presenting with dizziness to 45 general practitioners in the Netherlands from July 2006 to January 2008. We performed tests, including patient history, and physical and additional examination, previously selected by an international expert panel and based on an earlier systematic review. Our primary outcome was impact of dizziness on everyday life measured with the Dutch validated version of the Dizziness Handicap Inventory (DHI). After a bootstrap procedure (1500x) we investigated predictability of DHI-scores with stepwise backward multiple linear and logistic regressions. DHI-scores varied from 0 to 88 (maximum score: 100) and 60% of patients experienced moderate or severe impact on everyday life due to dizziness. Indicators for dizziness-related impairment were: onset of dizziness 6 months ago or more (OR 2.8, 95% CI 1.7-4.7), frequency of dizziness at least daily (OR 3.3, 95% CI 2.0-5.4), duration of dizziness episode one minute or less (OR 2.4, 95% CI 1.5-3.9), presence of anxiety and/or depressive disorder (OR 4.4, 95% CI 2.2-8.8), use of sedative drugs (OR 2.3, 95% CI 1.3-3.8) , and impaired functional mobility (OR 2.6, 95% CI 1.7-4.2). For this model with only 6 indicators the AUC was .80 (95% CI .76-.84). Dizziness-related impairment in older primary care patients is considerable (60%). With six simple indicators it is possible to identify which patients suffer the most from their dizziness without exactly knowing the cause(s) of their dizziness. Influencing these indicators, if possible, may lead to functional improvement and this might be effective in patients with moderate or severe impact of dizziness on their daily lives.


Rasch analysis of the School Function Assessment provides additional evidence for the internal validity of the activity performance scales.
The internal construct validity of the School Function Assessment (SFA) was determined by examining its unidimensionality and hierarchical structure. The SFA was completed for 64 elementary school children (35 with disabilities, 29 without disabilities). Data were examined through Rasch analysis. The majority of test items (252 of 266) within the Activity Performance Scales met the criterion set for Rasch goodness-of-fit statistics. All but three scales were found to be unidimensional, measuring a single construct. Item difficulty analysis yielded a hierarchical structure of the scales similar to the existing item layout of the SFA. Internal validity was supported for 15 of the 18 Activity Performance scales. Each scale is psychometrically sound in measuring a specific functional task. The reliable hierarchical pattern of the assessment helps anticipate and document the student's progress in functional performance from easier to more difficult school tasks.


Preparing a collection of radiology examinations for distribution and retrieval.
Clinical documents made available for secondary use play an increasingly important role in discovery of clinical knowledge, development of research methods, and education. An important step in facilitating secondary use of clinical document collections is easy access to descriptions and samples that represent the content of the collections. This paper presents an approach to developing a collection of radiology examinations, including both the images and radiologist narrative reports, and making them publicly available in a searchable database. The authors collected 3996 radiology reports from the Indiana Network for Patient Care and 8121 associated images from the hospitals' picture archiving systems. The images and reports were de-identified automatically and then the automatic de-identification was manually verified. The authors coded the key findings of the reports and empirically assessed the benefits of manual coding on retrieval. The automatic de-identification of the narrative was aggressive and achieved 100% precision at the cost of rendering a few findings uninterpretable. Automatic de-identification of images was not quite as perfect. Images for two of 3996 patients (0.05%) showed protected health information. Manual encoding of findings improved retrieval precision. Stringent de-identification methods can remove all identifiers from text radiology reports. DICOM de-identification of images does not remove all identifying information and needs special attention to images scanned from film. Adding manual coding to the radiologist narrative reports significantly improved relevancy of the retrieved clinical documents. The de-identified Indiana chest X-ray collection is available for searching and downloading from the National Library of Medicine (http://openi.nlm.nih.gov/).


The cost-effectiveness of radiofrequency catheter ablation as first-line treatment for paroxysmal atrial fibrillation: results from a MANTRA-PAF substudy.
The aim of this prospective substudy was to estimate the cost-effectiveness of treating paroxysmal atrial fibrillation (AF) with radiofrequency catheter ablation (RFA) compared with antiarrhythmic drugs (AADs) as first-line treatment. A decision-analytic Markov model, based on MANTRA-PAF (Medical Antiarrhythmic Treatment or Radiofrequency Ablation in Paroxysmal Atrial Fibrillation) study data, was developed to study long-term effects and costs of RFA compared with AADs as first-line treatment. Positive clinical effects were found in the overall population, a gain of an average 0.06 quality-adjusted life years (QALYs) to an incremental cost of €3033, resulting in an incremental cost-effectiveness ratio of €50 570/QALY. However, the result of the subgroup analyses showed that RFA was less costly and more effective in younger patients. This implied an incremental cost-effectiveness ratio of €3434/QALY in ≤50-year-old patients respectively €108 937/QALY in >50-year-old patients. Radiofrequency catheter ablation as first-line treatment is a cost-effective strategy for younger patients with paroxysmal AF. However, the cost-effectiveness of using RFA as first-line therapy in older patients is uncertain, and in most of these AADs should be attempted before RFA (MANTRA-PAF ClinicalTrials.gov number; NCT00133211).


Computerized visual feedback: an adjunct to robotic-assisted gait training.
Robotic devices for walking rehabilitation allow new possibilities for providing performance-related information to patients during gait training. Based on motor learning principles, augmented feedback during robotic-assisted gait training might improve the rehabilitation process used to regain walking function. This report presents a method to provide visual feedback implemented in a driven gait orthosis (DGO). The purpose of the study was to compare the immediate effect on motor output in subjects during robotic-assisted gait training when they used computerized visual feedback and when they followed verbal instructions of a physical therapist. Twelve people with neurological gait disorders due to incomplete spinal cord injury participated. Subjects were instructed to walk within the DGO in 2 different conditions. They were asked to increase their motor output by following the instructions of a therapist and by observing visual feedback. In addition, the subjects' opinions about using visual feedback were investigated by a questionnaire. Computerized visual feedback and verbal instructions by the therapist were observed to result in a similar change in motor output in subjects when walking within the DGO. Subjects reported that they were more motivated and concentrated on their movements when using computerized visual feedback compared with when no form of feedback was provided. Computerized visual feedback is a valuable adjunct to robotic-assisted gait training. It represents a relevant tool to increase patients' motor output, involvement, and motivation during gait training, similar to verbal instructions by a therapist.


Fife, a Drosophila Piccolo-RIM homolog, promotes active zone organization and neurotransmitter release.
Neuronal communication depends on the precisely orchestrated release of neurotransmitter at specialized sites called active zones (AZs). A small number of scaffolding and cytoskeletal proteins comprising the cytomatrix of the active zone (CAZ) are thought to organize the architecture and functional properties of AZs. The majority of CAZ proteins are evolutionarily conserved, underscoring the fundamental similarities in neurotransmission at all synapses. However, core CAZ proteins Piccolo and Bassoon have long been believed exclusive to vertebrates, raising intriguing questions about the conservation of the molecular mechanisms that regulate presynaptic properties. Here, we present the identification of a piccolo-rim-related gene in invertebrates, together with molecular phylogenetic analyses that indicate the encoded proteins may represent Piccolo orthologs. In accordance, we find that the Drosophila homolog, Fife, is neuronal and localizes to presynaptic AZs. To investigate the in vivo function of Fife, we generated a deletion of the fife locus. We find that evoked neurotransmitter release is substantially decreased in fife mutants and loss of fife results in motor deficits. Through morphological analysis of fife synapses, we identify underlying AZ abnormalities including pervasive presynaptic membrane detachments and reduced synaptic vesicle clustering. Our data demonstrate the conservation of a Piccolo-related protein in invertebrates and identify critical roles for Fife in regulating AZ structure and function. These findings suggest the CAZ is more conserved than previously thought, and open the door to a more complete understanding of how CAZ proteins regulate presynaptic structure and function through genetic studies in simpler model systems.


Organ donation after circulatory death: the forgotten donor?
Donation after circulatory death (DCD) can be performed on neurologically intact donors who do not fulfill neurologic or brain death criteria before circulatory arrest. This commentary focuses on the most controversial donor-related issues anticipated from mandatory implementation of DCD for imminent or cardiac death in hospitals across the USA. We conducted a nonstructured review of selected publications and websites for data extraction and synthesis. The recommended 5 min of circulatory arrest does not universally fulfill the dead donor rule when applied to otherwise neurologically intact donors. Scientific evidence from extracorporeal perfusion in circulatory arrest suggests that the procurement process itself can be the event causing irreversibility in DCD. Legislative abandonment of the dead donor rule to permit the recovery of transplantable organs is necessary in the absence of an adequate scientific foundation for DCD practice. The designation of organ procurement organizations or affiliates to obtain organ donation consent introduces self-serving bias and conflicts of interest that interfere with true informed consent. It is important that donors and their families are not denied a 'good death', and the impact of DCD on quality of end-of-life care has not been satisfactorily addressed to achieve this.


No ψ-epistemic model can fully explain the indistinguishability of quantum states.
According to a recent no-go theorem [M. Pusey, J. Barrett and T. Rudolph, Nat. Phys. 8, 475 (2012)], models in which quantum states correspond to probability distributions over the values of some underlying physical variables must have the following feature: the distributions corresponding to distinct quantum states do not overlap. In such a model, it cannot coherently be maintained that the quantum state merely encodes information about underlying physical variables. The theorem, however, considers only models in which the physical variables corresponding to independently prepared systems are independent, and this has been used to challenge the conclusions of that work. Here we consider models that are defined for a single quantum system of dimension d, such that the independence condition does not arise, and derive an upper bound on the extent to which the probability distributions can overlap. In particular, models in which the quantum overlap between pure states is equal to the classical overlap between the corresponding probability distributions cannot reproduce the quantum predictions in any dimension d ≥ 3. Thus any ontological model for quantum theory must postulate some extra principle, such as a limitation on the measurability of physical variables, to explain the indistinguishability of quantum states. Moreover, we show that as d→∞, the ratio of classical and quantum overlaps goes to zero for a class of states. The result is noise tolerant, and an experiment is motivated to distinguish the class of models ruled out from quantum theory.


Patients' Data Management System Protected by Identity-Based Authentication and Key Exchange.
A secure and distributed framework for the management of patients' information in emergency and hospitalization services is proposed here in order to seek improvements in efficiency and security in this important area. In particular, confidentiality protection, mutual authentication, and automatic identification of patients are provided. The proposed system is based on two types of devices: Near Field Communication (NFC) wristbands assigned to patients, and mobile devices assigned to medical staff. Two other main elements of the system are an intermediate server to manage the involved data, and a second server with a private key generator to define the information required to protect communications. An identity-based authentication and key exchange scheme is essential to provide confidential communication and mutual authentication between the medical staff and the private key generator through an intermediate server. The identification of patients is carried out through a keyed-hash message authentication code. Thanks to the combination of the aforementioned tools, a secure alternative mobile health (mHealth) scheme for managing patients' data is defined for emergency and hospitalization services. Different parts of the proposed system have been implemented, including mobile application, intermediate server, private key generator and communication channels. Apart from that, several simulations have been performed, and, compared with the current system, significant improvements in efficiency have been observed.


Insight into the structural requirements of benzothiadiazine scaffold-based derivatives as hepatitis C virus NS5B polymerase inhibitors using 3D-QSAR, molecular docking and molecular dynamics.
Hepatitis C virus (HCV) infection is a significant world health threat with frequently ineffective problem existed in the present treatment, thus representing a major unmet medical need. The nonstructural viral protein 5B (NS5B), one of the best-studied polymerase, has emerged as an attractive target for the development of novel therapeutics against hepatitis C virus. In this work, both ligand- and receptor- based three-dimensional quantitative structure activity relationship (3D-QSAR) studies were carried out using comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) techniques on 360 benzothiadiazine scaffold-based derivatives as HCV GT-1b NS5B polymerase allosteric inhibitors. The resultant optimum 3D-QSAR model exhibited R(2)(ev) of 0.54, R(2)(nev) of 0.72 and the predictive ability was validated by using an independent test set of 90 compounds which gave R(2)(pred) value of 0.64. In addition, docking analysis and molecular dynamics simulation (MD) were also applied to elucidate the probable binding modes of these inhibitors at the allosteric site of the enzyme. Interpretation of the 3D contour maps in context of the topology of the allosteric binding site of NS5B provided insight into NS5B-inhibitor interactions. The information obtained from this work can be utilized to accurately predict the binding affinity of related analogues and also facilitate the future rational design of novel inhibitors with improved activity.


Para-arytenoid cartilage pseudodiverticulum formation as a sequela of endotracheal intubation.
Laryngotracheal injury secondary to endotracheal intubation results from ischemic necrosis, which develops within hours of intubation, with injury severity increasing with duration of intubation. Postextubation and postmortem observations demonstrate that laryngotracheal injury still occurs in a large number of intubated patients. To demonstrate laryngeal injury, specifically pseudodiverticulum formation, as a result of intubation. A total of 41 autopsy cases from February 1970 to February 2006 were retrospectively reviewed. All patients had been intubated during their hospital course and had an endotracheal tube in place at the time of death. Continuous endotracheal intubation ranged from several hours to 33 days. The earliest manifestations of laryngeal injury occurred within hours and included hyperemia, edema, and ovoid mucosal lesions over the arytenoid and inner posterolateral cricoid cartilages. Within 48 hours, superficial epithelial denudation progressed to mucosal ulceration, with inflammatory infiltrate into the submucosa. These ulcerations enlarged and deepened, and within 3 to 6 days they showed gross excavations and, in some cases, confluence with other ulcerations. Four cases had pseudodiverticulum formation in the area of the arytenoid cartilages, with deep, excavated lesions that extended to the underlying cartilage. Pseudodiverticulum formation occurred as early as 8 days of continuous intubation. Although endotracheal intubation is required for airway maintenance in many patients with respiratory dysfunction, the procedure carries risks. Laryngeal injury from intubation is very common, and the severity of injury is directly related to the duration of intubation. We add para-arytenoid pseudodiverticulum formation to the list of possible complications of endotracheal intubation.


Fluorescence of 2-aminopurine reveals rapid conformational changes in the RB69 DNA polymerase-primer/template complexes upon binding and incorporation of matched deoxynucleoside triphosphates.
We have used 2-aminopurine (2AP) as a fluorescent probe in the template strand of a 13/20mer primer/template (D) to detect deoxynucleoside triphosphates (N)-dependent conformational changes exhibited by RB69 DNA polymerase (ED) complexes. The rates and amplitudes of fluorescence quenching depend hyperbolically on the [dTTP] when a dideoxy-primer/template (ddP/T) with 2AP as the templating base (n position) is used. No detectable fluorescence changes occur when a ddP/T with 2AP positioned 5' to the templating base (n + 1 position) is used. With a deoxy-primer/template (dP/T) with 2AP in the n position, a rapid fluorescence quenching occurs within 2 ms, followed by a second, slower fluorescence quenching with a rate constant similar to base incorporation as determined by chemical quench. With a dP/T having 2AP in the n + 1 position, there is a [dNTP]-dependent fluorescence enhancement that occurs at a rate comparable to dNMP incorporation. Collectively, the results favor a minimal kinetic scheme in which population of two distinct biochemical states of the ternary EDN complex precedes the nucleotidyl transfer reaction. Observed differences between dP/T and ddP/T ternary complexes indicate that the 3' hydroxyl group of the primer plays a critical role in determining the rate constants of transitions that lead to strong deoxynucleoside triphosphate binding prior to chemistry.


Dynamics of human milk nutrient composition of women from Singapore with a special focus on lipids.
A recent report suggested that human milk (HM) composition not only changes with lactation stages but also vary according to gender of the offspring. In spite of available literature, the dynamic changes of HM composition still remain to be completely explored and characterized. Progress in analytical technologies together with quantitative sampling of HM allows for a better quantification of HM nutrients and thereby providing a deeper understanding of the dynamics of HM secretion. To characterize and quantify HM nutrients based on appropriate for analyses sampling procedures and advanced analytical methodologies. We conducted an observatory, single center, longitudinal trial with HM collection at 30, 60, and 120 days postpartum from 50 mothers (singleton-deliveries of 25 male and 25 female infants). HM samples were analyzed for lipid, lactose, energy density, fatty acids, phospholipids, and gangliosides. Longitudinal analyses of the datasets have been carried out using linear mixed models. HM for male infants compared to females at 120 days, were higher for energy content and lipids by 24 and 39%, respectively. Similarly, other bioactive lipids such as linoleic acid, phospholipids and gangliosides were also significantly different based on the gender of the infant. Significant stage-based differences were observed for total lipids, energy density, phospholipids, and gangliosides. Such difference in HM composition may stem from different energy needs to cope up for individual growth and development. Collectively, the current observations affirm that HM secretion, especially the lipid composition, is a very dynamic and personalized biological process.


Decision support in nuclear emergencies.
In a nuclear emergency, protective actions such as evacuation, sheltering and food bans can be taken to mitigate the consequences of any release of radioactivity. Within the RODOS project, an evaluation framework has been developed to support the assessment of the costs and benefits of potential actions. In order to help the decision makers gain insight into the decision problem and clarify their preferences, guidance can be given in three stages. First, the search of feasible portfolios of protective actions is seen as a constraint satisfaction problem; only those portfolios that satisfy constraints depending on factors such as feasibility are worth further evaluation. Second, the portfolios are ranked based on their consequences and the preferences of the decision makers using either a multi-attribute value or utility function. Third, a natural language report explaining the ranking is produced to help the decision makers gain insight into the decision problem and refine the decision parameters. An intelligent decision system has been developed to demonstrate the feasibility of the framework.


Effects of mycotoxin-contaminated feed on farm animals.
Mycotoxins are secondary products produced by fungi in cereals and are frequently found in the livestock industry as contaminants of farm animal feed. Studies analyzing feed mycotoxins have been conducted worldwide and have confirmed the presence of mycotoxins with biological activity, including aflatoxin, ochratoxin A, fumonisin, zearalenone, and deoxynivalenol, in a large proportion of feed samples. Exposure to mycotoxins can cause immunotoxicity and impair reproductive function in farm animals. In addition, exposure of tissues, such as the kidneys, liver, and intestines, to mycotoxins can exert histopathological changes that can interfere with animal growth and survival. This review describes previous studies regarding the presence of major mycotoxins in the feed of farm animals, especially pigs and poultry. Moreover, it describes the adverse effects of mycotoxins in farm animals following exposure, as well as the biological activity of mycotoxins in animal-derived cells. Mycotoxins have been shown to regulate signaling pathways, oxidative stress, endoplasmic reticulum stress, apoptosis, and proliferation in porcine and bovine cells. A clear understanding of the effects of mycotoxins on farm animals will help reduce farm household economic loss and address the health concerns of people who consume these meat and dairy products.


Rolapitant for the prevention of postoperative nausea and vomiting: a prospective, double-blinded, placebo-controlled randomized trial.
Postoperative nausea and vomiting (PONV) are common complications after surgery. Neurokinin-1 (NK(1)) receptor antagonists have been shown to be safe and effective for the prevention and treatment of PONV in humans. Rolapitant is a potent, selective NK1 receptor antagonist that is rapidly absorbed, has a remarkably long half-life (up to180 hours), and appears to have a low potential for drug-drug interactions. We evaluated the dose response for rolapitant for the prevention of PONV in subjects at high risk for this condition, and rolapitant's effects on preventing delayed PONV were explored up to 5 days after surgery. A randomized, multicenter, double-blind, dose-ranging study of rolapitant was conducted with placebo and active control groups. Six hundred nineteen adult women undergoing open abdominal surgery were randomly assigned in equal ratios to 1 of 6 study arms: oral rolapitant in 5-mg, 20-mg, 70-mg, or 200-mg doses; IV ondansetron 4 mg; or placebo, stratified by history of PONV or motion sickness. The primary study endpoint was absence of emetic episodes, regardless of use of rescue medication, at 24 hours after extubation. Groups assigned to rolapitant 20-mg, 70-mg, and 200-mg had a higher incidence of no emesis in comparison with placebo at 24 hours after surgery. A linear relationship between rolapitant dose and primary outcome was seen. The probability of an emetic episode was significantly lower in the rolapitant 70-mg and 200-mg groups in comparison with placebo (P ≤ 0.001 based on the log-rank test). No significant differences were noted between rolapitant and the active control (ondansetron) at 24 hours after surgery, but there was a higher incidence of no emesis (regardless of rescue medication use) in the rolapitant 200- and 70-mg groups at 72 and 120 hours, respectively. Rolapitant is superior to placebo in reducing emetic episodes after surgery and reduces the incidence of vomiting in a dose-dependent manner. No differences in side effect profile were observed between rolapitant and placebo.


Polyamine permeation and rectification of Kir4.1 channels.
Inward rectifier K(+) (Kir) channels are expressed in multiple neuronal and glial cells. Recent studies have equated certain properties of exogenously expressed Kir4.1 channels with those of native K(+) currents in brain cells, as well as demonstrating the expression of Kir4.1 subunits in these tissues. There are nagging problems however with assigning native currents to Kir4.1 channels. One major concern is that in many native tissues, the putatively correlated currents show much weaker rectification than typically reported for cloned Kir4.1 channels. We have now examined the polyamine-dependence of Kir4.1 channels expressed at high density in Cosm6 cells, using inside-out membrane patches. The experiments reveal a complex and variable rectification that can help explain the variability reported for candidate Kir4.1 currents in native cells. Most importantly, rectification seems to be incomplete, even at high polyamine concentrations. In excised membrane patches, with high levels of expression, and high concentrations of spermine, there is approximately 15% residual conductance that is insensitive to spermine. From a biophysical perspective, this is a striking finding, and indicates either that a bound spermine fails to completely block permeation or that significant spermine permeation (i.e. 'punchthrough') is occurring. To examine this further, we have examined block by philanthotoxin (PhTx, essentially spermine with a bulky tail). PhTx block, while less potent, is more complete than spermine block. This leads us to propose that spermine 'punchthrough' may be significant in Kir4 channels, and that this may be a major contributor to the weak rectification observed under physiological conditions.


Hyperactive boys almost grown up. II. Status of subjects without a mental disorder.
In a previous article we reported the rate of DSM-III diagnoses among 101 male adolescents (aged 16 to 23 years) in whom hyperactivity had been diagnosed between ages 6 and 12 years compared with 100 controls. This report examines the rates of dysfunction among the 52 probands and 80 control adolescents who did not receive a DSM-III diagnosis at follow-up. Contrary to expectations, there were few areas (primarily involving school adjustment) in which the formerly hyperactive children were found to be at a disadvantage and numerous areas (occupational adjustment, temperament, alcohol abuse, antisocial activities, etc) in which the groups were indistinguishable. The results are explained in accordance with a bimodal distribution of dysfunction in hyperactive children grown up. Indirectly, this model supports the construct validity of the DSM-III diagnoses attention deficit disorder with hyperactivity, substance use disorder, and antisocial personality disorder. Post hoc analyses suggest that behavior problems resulting from drug use in early adolescence have graver consequences for previously hyperactive children than normal subjects.


Menopausal symptoms and ethnicity: the Study of Women's Health Across the Nation.
A number of studies have suggested that ethnic background influences a woman's perception of her symptoms. The Study of Women's Health Across the Nation (SWAN) is a multiethnic, longitudinal, cohort study of US women that includes non-Hispanic Caucasian, African-American, Chinese, Japanese and Hispanic women. The initial strategy for this seven-site study involved community-based recruitment of non-Hispanic Caucasians at each site, plus one minority ethnic group. Since ethnicity varies with many other factors, measures of education, acculturation, social status, psychological wellbeing and financial strain were all taken into account in interpreting symptom onset, frequency and severity of the common menopausal symptoms. Biological and physical measures were also assessed and related to symptoms. Most symptoms varied by ethnicity. Vasomotor symptoms were more prevalent in African-American and Hispanic women and were also more common in women with greater BMI, challenging the widely held belief that obesity is protective against vasomotor symptoms. Vaginal dryness was present in 30-40% of SWAN participants at baseline, and was most prevalent in Hispanic women. Among Hispanic women, symptoms varied by country of origin. Acculturation appears to play a complex role in menopausal symptomatology. We conclude that ethnicity should be taken into account when interpreting menopausal symptom presentation in women.


Pupil light reflex evoked by light-emitting diode and computer screen: Methodology and association with need for recovery in daily life.
Pupil light reflex (PLR) has been widely used as a method for evaluating parasympathetic activity. The first aim of the present study is to develop a PLR measurement using a computer screen set-up and compare its results with the PLR generated by a more conventional setup using light-emitting diode (LED). The parasympathetic nervous system, which is known to control the 'rest and digest' response of the human body, is considered to be associated with daily life fatigue. However, only few studies have attempted to test the relationship between self-reported daily fatigue and physiological measurement of the parasympathetic nervous system. Therefore, the second aim of this study was to investigate the relationship between daily-life fatigue, assessed using the Need for Recovery scale, and parasympathetic activity, as indicated by the PLR parameters. A pilot study was conducted first to develop a PLR measurement set-up using a computer screen. PLRs evoked by light stimuli with different characteristics were recorded to confirm the influence of light intensity, flash duration, and color on the PLRs evoked by the system. In the subsequent experimental study, we recorded the PLR of 25 adult participants to light flashes generated by the screen set-up as well as by a conventional LED set-up. PLR parameters relating to parasympathetic and sympathetic activity were calculated from the pupil responses. We tested the split-half reliability across two consecutive blocks of trials, and the relationships between the parameters of PLRs evoked by the two set-ups. Participants rated their need for recovery prior to the PLR recordings. PLR parameters acquired in the screen and LED set-ups showed good reliability for amplitude related parameters. The PLRs evoked by both set-ups were consistent, but showed systematic differences in absolute values of all parameters. Additionally, higher need for recovery was associated with faster and larger constriction of the PLR. This study assessed the PLR generated by a computer screen and the PLR generated by a LED. The good reliability within set-ups and the consistency between the PLRs evoked by the set-ups indicate that both systems provides a valid way to evoke the PLR. A higher need for recovery was associated with faster and larger constricting PLRs, suggesting increased levels of parasympathetic nervous system activity in people experiencing higher levels of need for recovery on a daily basis.


Zebra fish myc family and max genes: differential expression and oncogenic activity throughout vertebrate evolution.
To gain insight into the role of Myc family oncoproteins and their associated protein Max in vertebrate growth and development, we sought to identify homologs in the zebra fish (Brachydanio rerio). A combination of a polymerase chain reaction-based cloning strategy and low-stringency hybridization screening allowed for the isolation of zebra fish c-, N-, and L-myc and max genes; subsequent structural characterization showed a high degree of conservation in regions that encode motifs of known functional significance. On the functional level, zebra fish Max, like its mammalian counterpart, served to suppress the transformation activity of mouse c-Myc in rat embryo fibroblasts. In addition, the zebra fish c-myc gene proved capable of cooperating with an activated H-ras to effect the malignant transformation of mammalian cells, albeit with diminished potency compared with mouse c-myc. With respect to their roles in normal developing tissues, the differential temporal and spatial patterns of steady-state mRNA expression observed for each zebra fish myc family member suggest unique functions for L-myc in early embryogenesis, for N-myc in establishment and growth of early organ systems, and for c-myc in increasingly differentiated tissues. Furthermore, significant alterations in the steady-state expression of zebra fish myc family genes concomitant with relatively constant max expression support the emerging model of regulation of Myc function in cellular growth and differentiation.


Characterization of variation in the canine suppressor of cytokine signaling-2 (SOCS2) gene.
Suppressor of cytokine signaling 2 (SOCS2) is a negative regulator of growth hormone signaling. The deletion of SOCS2 in mice results in a 30-50% increase in post-natal growth. In an effort to identify polymorphisms in the SOCS2 gene that may be associated with body size in dogs, we characterized the canine SOCS2 gene and analyzed its genetic diversity among small and large dog breeds. The study was carried out on a total of 520 dogs from 66 different breeds. Dogs were classified as large or small based on height and weight as determined by their respective American Kennel Club breed standards. The SH2 and SOCS domains of the canine SOCS2 gene were sequenced in 32 dogs from different breeds. Only one non-synonymous sequence variant (DQ415457:g.326G>T) was detected which corresponds to an amino acid change (Asp127Tyr). All samples were genotyped by PCR/RFLP and the allele frequencies were determined for each dog breed. The T allele was distributed primarily among European large dog breeds with a gene frequency ranging from 0.72 to 0.04. The nature of the nucleotide change and the effect on the protein together with the finding of a QTL related to body size in the same CFA15 region by other researchers suggest canine SOCS2 as a potential candidate gene for body size in dogs. Future studies will be needed to clarify the role of the 326G>T polymorphism and its interaction with genes like growth hormone and insulin-like growth factor 1.


Does body mass index in childhood predict restraint eating in early adolescence?
The aim of this study was (1) to examine whether childhood BMI is a significant predictor of restrained eating in preadolescents, (2) to investigate gender differences in restrained and emotional eating, and (3) to determine whether emotional problems, and body esteem were related to eating problems of preadolescents. In this longitudinal study with two measurement points, data from 428 children (50% female) were used. At time 1 (t1) children were on average 5.9years old. BMI was assessed using objective measures. At time 2 (t2) participants were 12years old. The adolescents and their parents completed questionnaires assessing restrained and emotional eating, body esteem, emotional problems, and BMI. Multiple regression analysis showed that restrained eating was significantly predicted by t1 BMI, by change in BMI between t1 and t2, and t2 body esteem. Emotional eating was, as expected, not predicted by t1 BMI, but associated with t2 body esteem and t2 emotional problems. Gender was not a significant predictor. The stability of BMI between childhood and preadolescence and its ability to predict restrained eating suggests that it is important to start prevention of overweight, body dissatisfaction and disordered eating at an early age.


Quantification of rate constants for successive enzymatic reactions with DNP hyperpolarized MR.
A kinetic model is provided to obtain reaction rate constants in successive enzymatic reactions that are monitored using NMR spectroscopy and hyperpolarized substrates. The model was applied for simulation and analysis of the successive oxidation of choline to betaine aldehyde, and further to betaine, by the enzyme choline oxidase. This enzymatic reaction was investigated under two different sets of conditions: two different choline molecular probes were used, [1,1,2,2-D4 , 1-(13) C]choline chloride and [1,1,2,2-D4 , 2-(13) C]choline chloride, in different MR systems (clinical scanner and high-resolution spectrometer), as well as in different reactors and reaction volumes (4.8 and 0.7 mL). The kinetic analysis according to the model yielded similar results in both set-ups, supporting the robustness of the model. This was achieved despite the complex and negating influences of reaction kinetics and polarization decay, and in the presence of uncontrolled mixing characteristics, which may introduce uncertainties in both effective timing and effective pulses. The ability to quantify rate constants using hyperpolarized MR in the first seconds of consecutive enzyme activity is important for further development of the utilization of dynamic nuclear polarization-MR for biological determinations.


In pursuit of a better world: crop improvement and the CGIAR.
The CGIAR crop improvement (CI) programs, unlike commercial CI programs, which are mainly geared to profit though meeting farmers' needs, are charged with meeting multiple objectives with target populations that include both farmers and the community at large. We compiled the opinions from >30 experts in the private and public sector on key strategies, methodologies, and activities that could the help CGIAR meet the challenges of providing farmers with improved varieties while simultaneously meeting the goals of: (i) nutrition, health, and food security; (ii) poverty reduction, livelihoods, and jobs; (iii) gender equality, youth, and inclusion; (iv) climate adaptation and mitigation; and (v) environmental health and biodiversity. We review the crop improvement processes starting with crop choice, moving through to breeding objectives, production of potential new varieties, selection, and finally adoption by farmers. The importance of multidisciplinary teams working towards common objectives is stressed as a key factor to success. The role of the distinct disciplines, actors, and their interactions throughout the process from crop choice through to adoption by farmers is discussed and illustrated.


Calcium concentration dynamics produced by synaptic activation of CA1 hippocampal pyramidal cells.
The spatial and temporal dynamics of many electrophysiological and biochemical processes in nerve cells are in turn dependent on the concentration dynamics of the second messenger calcium. We have used microfluorimetry of the calcium indicator fura-2 (Grynkiewicz et al., 1985) to measure and characterize synaptically activated calcium changes in individual CA1 pyramidal cells contained within guinea pig hippocampal slices. One component of the calcium changes was largely produced by influx through voltage-dependent Ca2+ channels (VDCCs). It consisted of large transient accumulations in the proximal-apical and basal dendrites; the amplitude was smaller in the distal-apical dendrites and the soma. This spatial profile was insensitive to the method of cell activation: stimulation of inputs located at different positions on the dendritic tree as well as antidromic stimulation produced only slight modifications. This component was not blocked by the NMDA antagonist 5-amino-4-phosphonovalerate (AP5) (Collingridge et al., 1983), was greatly reduced by Cd2+, partially reduced by nifedipine, and was increased by Bay-K 8644, providing the evidence that it was largely produced by influx through VDCCs. Blocking postsynaptic Na+ channels with QX-314 greatly reduced the accumulation amplitude, and spatial differences between proximal-dendritic and distal-dendritic regions were less pronounced, suggesting that active sodium conductances contribute significantly to the spatial activation of calcium conductances. Residual spatial differences that persist in QX-314 experiments are consistent with the idea that VDCCs have decreased density on distal-apical dendrites. A second component of accumulation was induced by ionic currents through NMDA receptor channels. It was blocked by AP5, unaffected by QX-314, attenuated and slowed down by elevated calcium buffering, and spatially localized to regions receiving activated synaptic inputs. The magnitude of this component was strongly dependent on the frequency and amplitude of synaptic activation. At high frequency, it was generally very large, often saturating the fura-2 (> 2 microM). Measurements made with the indicator furaptra also showed large localized AP5-sensitive fluorescence changes. Our results suggest that in dendritic regions near activated input fibers calcium levels may reach 2-10 microM. In general, our measurements of calcium dynamics provide an experimental basis for evaluating the spatial distribution of calcium conductances, the spatial distribution of calcium-activated electrophysiological and biochemical processes, and the spatial uniformity of calcium buffering and removal systems in CA1 hippocampal pyramidal cells. The time course and amplitude of Ca2+ transients we measured suggest that activation of Ca(2+)-dependent conductances [e.g., IK(Ca)] will be markedly different for different cellular regions.(ABSTRACT TRUNCATED AT 400 WORDS)


A location- and sharpness-specific tactile electronic skin based on staircase-like nanowire patches.
Human skin can sense an external object in a location-specific manner, simultaneously recognizing whether it is sharp or blunt. Such tactile capability can be achieved in both natural and stretched states. It is impractical to mimic this tactile function of human skin by designing pixelated sensor arrays across our whole curvilinear human body. Here, we report a new tactile electronic skin sensor based on staircase-like vertically aligned gold nanowires (V-AuNWs). With a back-to-back linear or spiral assembly of two staircase structures into a single sensor, we are able to recognize pressure in a highly location-specific manner for both non-stretched and stretched states (up to 50% strain); with a concentric design on the fingertip, we can identify the sharpness of an external object. We believe that our strategy opens up a new route to highly specific second-skin-like tactile sensors for electronic skin (E-skin) applications.


Potential of cycloaddition reactions to generate cytotoxic metal drugs in vitro.
Severe general toxicity issues blight many chemotherapeutics utilized in the treatment of cancers, resulting in the need for more selective drugs able to exert their biological activity at only the required location(s). Toward this aim, we report the development of an organometallic ruthenium compound, functionalized through a η(6)-bound arene ligand with a bicyclononyne derivative, able to participate in strain-promoted cycloaddition reactions with tetrazines. We show that combination of the ruthenium compound with a ditetrazine in biological media results in the in situ formation of a dinuclear molecule that is more cytotoxic toward cancer cells than the starting mononuclear ruthenium compound and tetrazine components. Such an approach may be extended to in vivo applications to construct a cytotoxic metallodrug at a tumor site, providing a novel approach toward the turn-on cytotoxicity of metallodrugs in the treatment of cancer.