Probabilistic tractography recovers a rostrocaudal trajectory of connectivity variability in the human insular cortex.
The insular cortex of macaques has a wide spectrum of anatomical connections whose distribution is related to its heterogeneous cytoarchitecture. Although there is evidence of a similar cytoarchitectural arrangement in humans, the anatomical connectivity of the insula in the human brain has not yet been investigated in vivo. In the present work, we used in vivo probabilistic white-matter tractography and Laplacian eigenmaps (LE) to study the variation of connectivity patterns across insular territories in humans. In each subject and hemisphere, we recovered a rostrocaudal trajectory of connectivity variation ranging from the anterior dorsal and ventral insula to the dorsal caudal part of the long insular gyri. LE suggested that regional transitions among tractography patterns in the insula occur more gradually than in other brain regions. In particular, the change in tractography patterns was more gradual in the insula than in the medial premotor region, where a sharp transition between different tractography patterns was found. The recovered trajectory of connectivity variation in the insula suggests a relation between connectivity and cytoarchitecture in humans resembling that previously found in macaques: tractography seeds from the anterior insula were mainly found in limbic and paralimbic regions and in anterior parts of the inferior frontal gyrus, while seeds from caudal insular territories mostly reached parietal and posterior temporal cortices. Regions in the putative dysgranular insula displayed more heterogeneous connectivity patterns, with regional differences related to the proximity with either putative granular or agranular regions.


The measurement of executive function at age 5: psychometric properties and relationship to academic achievement.
This study examined the psychometric properties and criterion validity of a newly developed battery of executive function (EF) tasks for use in early childhood. The battery was included in the Family Life Project (FLP), a prospective longitudinal study of families who were oversampled from low-income and African American families at the birth of a new child (N = 1,292). Ninety-nine percent (N = 1,036) of children who participated in the age 5 home visit completed 1 or more (M = 5.8, Mdn = 6) of the 6 EF tasks. Results indicated that tasks worked equally well for children residing in low-income and not low-income homes, that task scores were most informative about the ability level of children in the low-average range, that performance on EF tasks was best characterized by a single factor, and that individual differences on the EF battery were strongly related to a latent variable measuring overall academic achievement, as well as to individual standardized tests that measured phonological awareness, letter-word identification, and early math skills.


Biochemical modulation of 5-fluorouracil with murine interferon-alpha/beta against murine renal cell carcinoma.
Conventional therapy for renal cell carcinoma using interferon (IFN) has shown limited antitumor action. The purpose of our study was to investigate synergistic antitumor effects of IFN and 5-fluorouracil (5-FU), and to elucidate the mechanisms of interaction between the 2 agents in mice. Antitumor effects and biochemical modulation of murine IFN-alpha/beta and 5-FU were determined against the murine renal cell carcinoma cell line, Renca, in vivo. The activity of thymidylate synthetase and thymidine kinase was measured using cytosolic extracts of the tumors. Combination treatment with IFN-alpha/beta and 5-FU produced a significant enhancement of growth inhibition against Renca tumor. Treatment with 5-FU resulted in a 2.7-fold increase in the total amount of thymidylate synthetase and an 11.6-fold increase in the thymidylate synthetase inhibition rate, while the administration of IFN-alpha/beta did not significantly reduce the 5-FU-induced increase in thymidylate synthetase. The administration of IFN-alpha/beta decreased thymidine kinase activity to 65.5% maximally, compared with that in the control mice or the mice treated with 5-FU. The reduction of thymidine kinase caused by treating the mice with IFN-alpha/beta changes the utilization of exogenous thymidine for DNA synthesis, and may represent the mechanism of the additive antitumor effect of the 2 agents, through the suppression of the salvage pathway for deoxythymidine monophosphate induction.


Trends in addiction treatment in Irish prisons using national surveillance data, 2009-2014.
Many studies show that incarcerated populations have higher rates of problem drug use than the general population. The purpose of this paper is to analyse trends in addiction treatment demand in prisons in Ireland from 2009 to 2014 using available national surveillance data in order to identify any implications for practice and policy. National surveillance data on treatment episodes for problem drug and alcohol use from 2009 to 2014, collected annually by the National Drug Treatment Reporting System (NDTRS), were analysed. In total, 6 per cent of all treatment episodes recorded by the NDTRS between 2009 and 2014 were from prison services. The number of prison service treatment episodes increased from 964 in 2009 to 1,063 in 2014. Opiates were the main reason for treatment, followed by alcohol, cocaine and cannabis. The majority (94-98 per cent) of treatment episodes involved males (median age of 29 years) and low educational attainment, with 79.5-85.1 per cent leaving school before completion of second level. The percentage of treatment episodes with a history of ever injecting drugs increased from 20.9 per cent in 2009 to 31.0 per cent in 2014. This study can help policy development and service planning in addiction treatment in prison as it provides an insight into the potential needs of incarcerated populations. It also provides a baseline from which to measure any changes in provision of treatment in prison over time. This is the first study to analyse treatment episodes in prison using routine surveillance data in Ireland. Analysis of these data can provide useful information, not currently available elsewhere.


Differentiable neural substrates for learned and described value and risk.
Studies of human decision making emerge from two dominant traditions: learning theorists [1-3] study choices in which options are evaluated on the basis of experience, whereas behavioral economists and financial decision theorists study choices in which the key decision variables are explicitly stated. Growing behavioral evidence suggests that valuation based on these different classes of information involves separable mechanisms [4-8], but the relevant neuronal substrates are unknown. This is important for understanding the all-too-common situation in which choices must be made between alternatives that involve one or another kind of information. We studied behavior and brain activity while subjects made decisions between risky financial options, in which the associated utilities were either learned or explicitly described. We show a characteristic effect in subjects' behavior when comparing information acquired from experience with that acquired from description, suggesting that these kinds of information are treated differently. This behavioral effect was reflected neurally, and we show differential sensitivity to learned and described value and risk in brain regions commonly associated with reward processing. Our data indicate that, during decision making under risk, both behavior and the neural encoding of key decision variables are strongly influenced by the manner in which value information is presented.


Insights into the gate of the nuclear pore complex.
Nuclear pore complexes (NPCs) serve as the gateway of the cell nucleus. These macromolecular assemblies form selective aqueous translocation channels permitting the free diffusion of small molecules, as well as receptor-mediated transport of large cargoes. Over the past decade, major progress has been made in both the structural determination of individual nucleoporins and subcomplexes by X-ray crystallography and in the structural analysis of the entire NPC by cryo-electron tomography (cryo-ET). The metazoan NPC structure from Xenopus laevis oocytes was recently resolved up to 20 Å by combining cryo-ET with advanced image processing techniques, revealing for the first time the architecture of the central channel. Here, we discuss the structure of the Xenopus laevis NPC and consider future perspectives that will eventually allow reconstructing the scaffold and gate of the NPC with higher resolution and identifying its transport-relevant regions. This will eventually allow us to describe the structure of the NPC 'in action'.


Changes Induced by Mind-Body Intervention Including Epigenetic Marks and Its Effects on Diabetes.
Studies have evidenced that epigenetic marks associated with type 2 diabetes (T2D) can be inherited from parents or acquired through fetal and early-life events, as well as through lifelong environments or lifestyles, which can increase the risk of diabetes in adulthood. However, epigenetic modifications are reversible, and can be altered through proper intervention, thus mitigating the risk factors of T2D. Mind-body intervention (MBI) refers to interventions like meditation, yoga, and qigong, which deal with both physical and mental well-being. MBI not only induces psychological changes, such as alleviation of depression, anxiety, and stress, but also physiological changes like parasympathetic activation, lower cortisol secretion, reduced inflammation, and aging rate delay, which are all risk factors for T2D. Notably, MBI has been reported to reduce blood glucose in patients with T2D. Herein, based on recent findings, we review the effects of MBI on diabetes and the mechanisms involved, including epigenetic modifications.


Implementation of Telehealth Services at the US Department of Veterans Affairs During the COVID-19 Pandemic: Mixed Methods Study.
At the onset of the COVID-19 pandemic, there was a rapid increase in the use of telehealth services at the US Department of Veterans Affairs (VA), which was accelerated by state and local policies mandating stay-at-home orders and restricting nonurgent in-person appointments. Even though the VA was an early adopter of telehealth in the late 1990s, the vast majority of VA outpatient care continued to be face-to-face visits through February 2020. We compared telehealth service use at a VA Medical Center, Greater Los Angeles across 3 clinics (primary care [PC], cardiology, and home-based primary care [HBPC]) 12 months before and 12 months after the onset of COVID-19 (March 2020). We used a parallel mixed methods approach including simultaneous quantitative and qualitative approaches. The distribution of monthly outpatient and telehealth visits, as well as telephone and VA Video Connect encounters were examined for each clinic. Semistructured telephone interviews were conducted with 34 staff involved in telehealth services within PC, cardiology, and HBPC during COVID-19. All audiotaped interviews were transcribed and analyzed by identifying key themes. Prior to COVID-19, telehealth use was minimal at all 3 clinics, but at the onset of COVID-19, telehealth use increased substantially at all 3 clinics. Telephone was the main modality of patient choice. Compared with PC and cardiology, video-based care had the greatest increase in HBPC. Several important barriers (multiple steps for videoconferencing, creation of new scheduling grids, and limited access to the internet and internet-connected devices) and facilitators (flexibility in using different video-capable platforms, technical support for patients, identification of staff telehealth champions, and development of workflows to help incorporate telehealth into treatment plans) were noted. Technological issues must be addressed at the forefront of telehealth evolution to achieve access for all patient populations with different socioeconomic backgrounds, living situations and locations, and health conditions. The unprecedented expansion of telehealth during COVID-19 provides opportunities to create lasting telehealth solutions to improve access to care beyond the pandemic.


Effect of computer use in the consultation on the delivery of care.
The effects of the use of a computer on the delivery of care in consultations in general practice were examined. In this trial a computer system provided for the review and update of patients' medical histories, notes on doctor-patient contacts, and information on repeat prescribing. Thirty consultations in which the computer system was used and 30 consultations in which no computer was used were matched individually for the doctor consulted, the sex and age of the patient, and the presenting problem(s). Six independent general practitioners blind rated each consultation for the standard of care attained. A minor negative effect of computer use on the doctors' clinical performance was found. We suggest, however, that this small effect would disappear if a computer system was used routinely.


Topographical analysis of the optic nerve in migraine patients.
Vascular dysfunction appears related to the development of migraines and has been associated with pressure-independent glaucoma. The purpose of this study is to investigate possible topographical differences in the optic nerve between migraine sufferers and normal age-matched control subjects. The identification of optic disc topographical differences between migraine and non-migraine sufferers may help clinicians determine if the presence of migraine influences the development and progression of glaucoma. Sixty consecutively seen patient-volunteers (30 subjects in migraine group: mean age 37.7 years, range from 19 to 66 years, 28 females and 2 males; 30 subjects in the control group: mean age 37.6 years, range from 19 to 61 years, 29 females and 1 male) were categorized as either migraine patients or control group subjects. All migraine subjects had been medically diagnosed with a migraine syndrome and were being treated with prophylactic medication. Optic nerve head topographical analysis was performed using a confocal scanning laser ophthalmoscope. The data was evaluated using generalized estimating equations, a priori and post hoc power analysis. No significant differences were found in any of the optic nerve parameters examined between the two groups. This study indicates that the optic nerve parameters as measured by the Heidelberg retinal tomograph II do not differ between migraine sufferers and age-matched control subjects.


Polymorphic regulation of mitochondrial fission and fusion modifies phenotypes of microglia in neuroinflammation.
Microglia are the resident macrophages of the central nervous system and play complex roles in the milieu of diseases including the primary diseases of myelin. Although mitochondria are critical for cellular functions and survival in the nervous system, alterations in and the roles of mitochondrial dynamics and associated signaling in microglia are still poorly understood. In the present study, by combining immunohistochemistry and 3D ultrastructural analyses, we show that mitochondrial fission/fusion in reactive microglia is differentially regulated from that in monocyte-derived macrophages and the ramified microglia of normal white matter in myelin disease models. Mouse cerebral microglia in vitro demonstrated that stimulation of TLR4 with lipopolysaccharide, widely used to examine microglial reactions, caused the activation of the mitochondrial fission protein, dynamin-related protein 1 (Drp1) and enhanced production of reactive oxygen species (ROS). The increase in the ROS level activated 5' adenosine monophosphate-activated protein kinase (AMPK), and facilitated elongation of mitochondria along the microtubule tracks. These results suggest that the polymorphic regulation of mitochondrial fission and fusion in reactive microglia is mediated by distinct signaling under inflammatory conditions, and modulates microglial phenotypes through the production of ROS.


Comparative characterization of GmSPX members reveals that GmSPX3 is involved in phosphate homeostasis in soybean.
Proteins containing the SPX (SYG1/Pho81/XPR1) domain are vital components in the phosphorus (P) signalling pathway, and regulate phosphate (Pi) homeostasis in plants. However, the characteristics and functions of GmSPX members in soybean (Glycine max) remain largely unknown. BLAST searching revealed nine GmSPX members in the soybean genome. Subsequently, expression patterns of GmSPX members were investigated in various tissues of soybean grown in nutrient solution or sand culture through quantitative real-time PCR (qPCR) analysis. Sub-cellular localization of GmSPX was examined via transient expression of 35S:GmSPX-GFP in epidermal cells of onion (Allium cepa). Finally, soybean transgenic composite plants were generated to study GmSPX3 functions. Nine GmSPX members were identified, which were classified into three groups based on phylogenetic analysis. Diverse responses of GmSPX members to deficiencies of nutrients (nitrogen, phosphorus, potassium and iron) or inoculation of arbuscular mycorrhizal fungi and rhizobia were observed in soybean. In addition, variations of sub-cellular localization of GmSPX members were found. Among them, GmSPX3, GmSPX7 and GmSPX8 were localized in the nuclei, and the other GmSPX members were confined to the nuclei and cytoplasm. The nuclear-localized and Pi starvation responsive-gene, GmSPX3, was functionally analysed in soybean transgenic composite plants. Overexpression of GmSPX3 led to increased P concentrations in both shoots and roots in the high-P treatment, and increased transcription of seven Pi starvation-responsive genes in soybean hairy roots. Taken together, the results suggest that GmSPX3 is a positive regulator in the P signalling network, and controls Pi homeostasis in soybean.


Localization of matrix metalloproteinase 3 (stromelysin) in osteoarthritic cartilage and synovium.
Degradation of proteoglycans is an initial change in osteoarthritic cartilage. Matrix metalloproteinase-3 (MMP-3; stromelysin) capable of degrading cartilage proteoglycans and type IX collagen was immunolocalized in osteoarthritic and normal cartilage. Immunohistochemical studies showed MMP-3 in chondrocytes of the superficial and transition zones in approximately 90% of osteoarthritic cartilage (60 of 67 samples) and in 31% of those of the superficial zone in some normal cartilage (4 of 13 samples). MMP-3 staining correlated directly with the histological histochemical scores of Mankin and with proteoglycan depletion, up to a certain grade of severity. Chondrocytes in the deep radial zone, clusters, and osteophytes were immunostained only when proteoglycan depletion and fissures affected them. Culture media from osteoarthritic cartilage contained significantly higher levels of metalloproteinase activity that was identified as MMP-3 by immunoblotting and lower amounts of tissue inhibitor of metalloproteinases compared with those in the control samples. MMP-3 was also immunolocalized in the lining cells of most osteoarthritic synovium (20 of 23 specimens, 87%) with a direct correlation with scores of inflammatory cell infiltration in the synovium, but it was not detected in the normal synovium. Light and electron microscopic studies demonstrated that MMP-3 digests proteoglycan aggregates in human articular cartilage. Treatment of normal and osteoarthritic cartilage slices with tumor necrosis factor-alpha and/or interleukin-1 alpha increased the number of MMP-3-immunoreactive chondrocytes and the intensity of the staining. These data suggest that MMP-3 produced by the chondrocytes and synovial lining cells under stimulation with these cytokines may be important in proteoglycan degradation in human ostoearthritic cartilage.


Can diffusion-weighted imaging add information in the evaluation of breast lesions considered suspicious on magnetic resonance imaging?
To assess the role of diffusion-weighted imaging (DWI) in the evaluation of breast lesions classified as suspicious on magnetic resonance imaging (MRI), correlating the findings with the results of the histological analysis. This was a retrospective, descriptive study based on a review of the medical records of 215 patients who were submitted to MRI with DWI before undergoing biopsy at a cancer center. Apparent diffusion coefficient (ADC) values were calculated for each lesion, and the result of the histological analysis was considered the gold standard. The mean age was 49 years. We identified 252 lesions, 161 (63.9%) of which were found to be malignant in the histological analysis. The mean ADC value was higher for the benign lesions than for the malignant lesions (1.50 × 10-3 mm2/s vs. 0.97 × 10-3 mm2/s), the difference being statistically significant (p < 0.001). The ADC cut-off point with the greatest sensitivity and specificity on the receiver operating characteristic curve was 1.03 × 10-3 mm2/s. When the DWI and conventional MRI findings were combined, the accuracy reached 95.9%, with a sensitivity of 95.7% and a specificity of 96.4%. The use of DWI could facilitate the characterization of breast lesions, especially those classified as BI-RADS 4, increasing the specificity and diagnostic accuracy of MRI.


Hypoxia impairs skin myofibroblast differentiation and function.
Ischemic wounds are characterized by oxygen levels lower than that of healthy skin (hypoxia) and poor healing. To better understand the pathophysiology of impaired wound healing, we investigated how switching from high (21%) to low (2%) oxygen levels directly affects cultured skin myofibroblasts, essential cells for the normal wound repair process. Myofibroblast differentiation and function were assessed by quantifying α-smooth muscle actin expression and cell contraction in collagen gels and on wrinkling silicone substrates. Culture for 5 days at 2% oxygen is perceived as hypoxia and significantly reduced myofibroblast differentiation and contraction despite high levels of the profibrotic transforming growth factor-β1. Analysis of α-smooth muscle actin expression on wrinkling substrates over time showed that reduced myofibroblast contraction preceded α-smooth muscle actin disassembly from stress fibers after switching from 21 to 2% oxygen. These effects were reversible by restoring high oxygen conditions and by applying mechanical stress. We suggest that mechanical challenge is a clinical relevant strategy to improve ischemic and chronic wound healing by supporting myofibroblast formation.


Immunogenicity and adverse events of avian influenza A H5N1 vaccine in healthy adults: multiple-treatments meta-analysis.
Influenza H5N1 is thought to be a likely causative agent for a future human influenza pandemic. Several types of H5N1 vaccine have been tested, including different doses and adjuvants, and a meta-analysis is needed to identify the best formulation. We searched Medline, Embase, the Cochrane Library, and other online databases to February, 2009, in any language for randomised trials comparing different H5N1 vaccines with or without placebo in healthy adults. Primary outcomes were seroconversion, seroresponse, or both according to haemagglutination-inhibition and microneutralisation. Secondary outcomes were adverse events. Because of the large number of compared formulations, multiple-treatments meta-analysis was used for primary outcomes. Direct-comparison meta-analyses were also done. We included 13 trials, which assessed 58 groups. With non-aluminium adjuvant, sufficiently high immunogenicity (greater than 70%) was achieved even at 12 microg or less (given as two doses of 6 microg or less), and higher doses did not provide major improvements. Immunogenicity for non-adjuvanted and aluminium-adjuvanted formulations increased with increasing dose, but was not sufficiently high. No serious vaccine-related adverse events were reported across 9600 participants. Currently, H5N1 influenza vaccines with non-aluminium adjuvants might represent the best available option in a pandemic. Large-scale studies are needed to verify the high immunogenicity of non-aluminium-adjuvanted vaccines that use very low doses of antigen.


Seedless synthesis and SERS characterization of multi-branched gold nanoflowers using water soluble polymers.
We report for the first time, the aqueous-based synthesis of multibranched, monodispersed gold nanoflowers (AuNFs) using pyromellitic dianhydride-p-phenylene diamine - PPDDs at room temperature. AuNF synthesis was achieved using PPDDs that converts Au precursor (Au3+) into AuNFs while serving as both the reducing and directional agent. The resulting branched AuNFs exhibited different degrees of anisotropy and protuberance lengths obtained by modulating the ratio of PPDDs and HAuCl4·3H2O. The surface roughness obtained ranged from small bud-like protuberances to elongated spikes, which enabled the tuning of the optical properties of the nanoparticles from ∼450 to 1100 nm. Systematic analysis revealed that the generation of urchin-like particles as well as their size depended on the PPDDs/HAuCl4·3H2O ratio. At a medium concentration of the precursor, spherical nanoparticles were formed. Whereas at lower precursor concentrations, urchin-like nanoparticles were obtained with their size and protuberances length increasing at even lower HAuCl4·3H2O concentration. Increasing the temperature to 100 °C resulted in the enhancement of the anisotropy of the AuNFs. The resulting gold nanoflowers exhibited an enhanced performance in surface-enhanced Raman scattering (SERS). This work provides a unique approach for anisotropic particle synthesis using water soluble polymer and greener approaches. The fabricated AuNFs exhibited variable UV-vis absorption and SERS enhancement as a function of branch morphology, indicating their potential application in biolabeling, biosensing, imaging, and therapeutic applications.


Cloning and expression of a new member of the L-2-amino-4-phosphonobutyric acid-sensitive class of metabotropic glutamate receptors.
Despite the cloning of several metabotropic glutamate receptors (mGluR1-6), the activity and localization of the cloned mGluRs do not account for the action of L-2-amino-4-phosphonobutyric acid (L-AP4) on mitral/tufted cells in the rat olfactory bulb. Thus, we screened a rat olfactory bulb library for novel cDNA clones, using probes derived from mGluR1 and mGluR4. A full length cDNA clone encoding a metabotropic receptor (mGluR7) whose sequence was 69% identical to that of mGluR4 was isolated. Stimulation of mGluR7 with L-AP4 and glutamate (each at 1 mM) in stably transfected baby hamster kidney cells inhibited forskolin-stimulated cAMP formation, whereas ACPD (1 mM) and quisqualate (0.5 mM) were less effective. Inhibition of cAMP required high concentrations of agonist in the transfected cells, suggesting that inhibition of adenylate cyclase may not be the predominant transduction mechanism for this receptor in neurons. RNA blot analysis and in situ hybridization revealed that mGluR7 has an expression pattern in the central nervous system distinct from that of other L-AP4-sensitive mGluRs. Double-labeling with probes for mGluR1 and mGluR7 revealed that individual mitral/tufted neurons in the olfactory bulb expressed both mRNAs. The expression pattern and L-AP4 sensitivity of mGluR7 suggest that it mediates inhibition of transmitter release at selected glutamatergic synapses. The coexpression of multiple mGluR mRNAs in single neurons indicates that the cellular effects of mGluR activation are likely to result from the integrated action of several receptor subtypes.


A maternal-effect selfish genetic element in Caenorhabditis elegans.
Selfish genetic elements spread in natural populations and have an important role in genome evolution. We discovered a selfish element causing embryonic lethality in crosses between wild strains of the nematode Caenorhabditis elegans The element is made up of sup-35, a maternal-effect toxin that kills developing embryos, and pha-1, its zygotically expressed antidote. pha-1 has long been considered essential for pharynx development on the basis of its mutant phenotype, but this phenotype arises from a loss of suppression of sup-35 toxicity. Inactive copies of the sup-35/pha-1 element show high sequence divergence from active copies, and phylogenetic reconstruction suggests that they represent ancestral stages in the evolution of the element. Our results suggest that other essential genes identified by genetic screens may turn out to be components of selfish elements.


Adenylyl cyclase 5-generated cAMP controls cerebral vascular reactivity during diabetic hyperglycemia.
Elevated blood glucose (hyperglycemia) is a hallmark metabolic abnormality in diabetes. Hyperglycemia is associated with protein kinase A (PKA)-mediated stimulation of L-type Ca2+ channels in arterial myocytes resulting in increased vasoconstriction. However, the mechanisms by which glucose activates PKA remain unclear. Here, we showed that elevating extracellular glucose stimulates cAMP production in arterial myocytes, and that this was specifically dependent on adenylyl cyclase 5 (AC5) activity. Super-resolution imaging suggested nanometer proximity between subpopulations of AC5 and the L-type Ca2+ channel pore-forming subunit CaV1.2. In vitro, in silico, ex vivo and in vivo experiments revealed that this close association is critical for stimulation of L-type Ca2+ channels in arterial myocytes and increased myogenic tone upon acute hyperglycemia. This pathway supported the increase in L-type Ca2+ channel activity and myogenic tone in two animal models of diabetes. Our collective findings demonstrate a unique role for AC5 in PKA-dependent modulation of L-type Ca2+ channel activity and vascular reactivity during acute hyperglycemia and diabetes.


Dependence of signal on depth in transmission Raman spectroscopy.
Recently, transmission Raman spectroscopy has been shown to be a valuable tool in the volumetric quantification of pharmaceutical formulations. In this work a Monte Carlo simulation and experimental study are performed to elucidate the dependence of the Raman signal on depth from the viewpoint of probing pharmaceutical tablets and powders in this experimental configuration. The transmission Raman signal is shown to exhibit a moderate bias toward the center of the tablets and this can be considerably reduced by using a recently developed Raman signal-enhancing concept, the "photon diode." The enhancing element not only reduces the bias but also increases the overall Raman signal intensity and consequently improves the signal-to-noise ratio of the measured spectrum. Overall, its implementation with appropriately chosen reflectivity results in a more uniform volumetric sampling across the half of the tablet where the photon diode is used (or across the tablet's entire depth if two photon diodes are used on each side of tablet) and enhanced overall sensitivity. These findings are substantiated experimentally on a segmented tablet by inserting a poly(ethelyne terephthalate) (PET) film doped with TiO(2) at different depths and monitoring its contribution to the overall transmission Raman signal from the segmented tablet. The numerical simulations also indicate considerable sensitivity of the overall Raman signal to the absorption of the sample, which is in line with large migration distances traversed by photons in these measurements. The presence of sample absorption was shown numerically to reduce the signal enhancement effect while the overall depth-dependence profile remained broadly unchanged. The absorption was also shown to produce a depth profile with the photon diode similar to that without it, although with a reduced absolute intensity of Raman signals and diminished enhancement effect.


IL-1α signaling is critical for leukocyte recruitment after pulmonary Aspergillus fumigatus challenge.
Aspergillus fumigatus is a mold that causes severe pulmonary infections. Our knowledge of how A. fumigatus growth is controlled in the respiratory tract is developing, but still limited. Alveolar macrophages, lung resident macrophages, and airway epithelial cells constitute the first lines of defense against inhaled A. fumigatus conidia. Subsequently, neutrophils and inflammatory CCR2+ monocytes are recruited to the respiratory tract to prevent fungal growth. However, the mechanism of neutrophil and macrophage recruitment to the respiratory tract after A. fumigatus exposure remains an area of ongoing investigation. Here we show that A. fumigatus pulmonary challenge induces expression of the inflammasome-dependent cytokines IL-1β and IL-18 within the first 12 hours, while IL-1α expression continually increases over at least the first 48 hours. Strikingly, Il1r1-deficient mice are highly susceptible to pulmonary A. fumigatus challenge exemplified by robust fungal proliferation in the lung parenchyma. Enhanced susceptibility of Il1r1-deficient mice correlated with defects in leukocyte recruitment and anti-fungal activity. Importantly, IL-1α rather than IL-1β was crucial for optimal leukocyte recruitment. IL-1α signaling enhanced the production of CXCL1. Moreover, CCR2+ monocytes are required for optimal early IL-1α and CXCL1 expression in the lungs, as selective depletion of these cells resulted in their diminished expression, which in turn regulated the early accumulation of neutrophils in the lung after A. fumigatus challenge. Enhancement of pulmonary neutrophil recruitment and anti-fungal activity by CXCL1 treatment could limit fungal growth in the absence of IL-1α signaling. In contrast to the role of IL-1α in neutrophil recruitment, the inflammasome and IL-1β were only essential for optimal activation of anti-fungal activity of macrophages. As such, Pycard-deficient mice are mildly susceptible to A. fumigatus infection. Taken together, our data reveal central, non-redundant roles for IL-1α and IL-1β in controlling A. fumigatus infection in the murine lung.


Direct methods for studying transcription regulatory proteins and RNA polymerase in bacteria.
Transcription factors and sigma factors play a major role in bacterial gene regulation by guiding the distribution of RNA polymerase between the promoters of different transcription units in response to changes in the environment. For 40 years Escherichia coli K-12 has been the paradigm for investigating this regulation and most studies have focused on small numbers of promoters studied by a combination of genetics and biochemistry. Since the first complete sequence for a bacterial genome was reported, the emphasis has switched to studying transcription on a global scale, with transcriptomics and bioinformatics becoming the methods of choice. Here we discuss two complementary direct experimental methods for studying transcription factors and sigma factors and we outline their potential use in rapidly establishing the regulatory networks in newly sequenced bacteria.


Expenditure on obstetric care and the protective effect of insurance on the poor: lessons from two Indonesian districts.
The Indonesian Government recently introduced a health insurance scheme to improve access to care for the poor. We investigated the payments made by households for different types of obstetric care, the economic consequences of payments and the effects of the new insurance on that expenditure. Expenditures on obstetric care for women were collected from three main hospitals in the Serang and Pandeglang districts of Banten Province for all 'near-miss' cases (372), a sample of normal deliveries (146) and deliveries with Caesarean section (98) over a 6-month period. Women were also interviewed after they were discharged to collect information on economic status, household expenditure and source of payment for care. Average expenditure by the mothers for near-miss cases was found to be Rp 2.6m (US$279) and Rp 1.9m (US$205) in Serang and Pandeglang Hospitals, respectively. Caesarean section was found to be the most expensive intervention. Insurance for the poor covered 51% of women at Serang Hospital and 73% of women at Pandeglang. Around 68% of households in the poorest quintiles would have made catastrophic payments. Insurance for the poor appears to have some positive association with the hospitals' expenditure for treatment of different types of maternal care. Insurance for the poor appeared to be relatively effective in protecting households from catastrophic payments. However, it is not sufficient only to cover the very poor; the non-poor can also suffer catastrophic payments and they are only protected because hospital rules over who qualifies have been relaxed. Although the association between insurance and expenditure for obstetric care was important, it was not clear that this represents over-provision of services but rather that it reflected previously inadequate treatment given to those unable to pay.


Confirmatory non-invasive and non-destructive differentiation between hemp and cannabis using a hand-held Raman spectrometer.
Cannabis is a generic term that is used to denote hemp plants (Cannabis sativa) that produce delta-9-tetrahydrocannabinolic acid (THCA) in amounts higher than industrial hemp. While THCA itself is not considered psychoactive, it is the source of the psychoactive delta-9 tetrahydrocannabinol (THC) that forms from its oxidation. About 147 million people, which is around 2.5% of the world population, consume cannabis. This makes cannabis by far the most widely cultivated and trafficked illicit drug in the world. Such enormous popularity of cannabis requires substantial effort by border control and law enforcement agencies to control illegal trafficking and distribution. Confirmatory diagnostics of cannabis is currently done by high pressure liquid chromatography (HPLC), which requires sample transportation to a certified laboratory, making THC diagnostics extremely time and labor consuming. This catalyzed a push towards development of a portable, confirmatory, non-invasive and non-destructive approach for cannabis diagnostics that could be performed by a police officer directly in the field to verify illicit drug possession or transport. Raman spectroscopy (RS) is a modern analytical technique that meets all these strict expectations. In this manuscript, we show that RS can be used to determine whether plant material is hemp or cannabis with 100% accuracy. We also demonstrate that RS can be used to probe the content of THCA in the analyzed samples. These findings suggest that a hand-held Raman spectrometer can be an ideal tool for police officers and hemp breeders to enable highly accurate diagnostics of THCA content in plants.


The effects of different types of music on mood, tension, and mental clarity.
This study investigated the impact of different types of music on tension, mood, and mental clarity. A total of 144 subjects completed a psychological profile before and after listening for 15 minutes to four types of music (grunge rock, classical, New Age, and designer). With grunge rock music, significant increases were found in hostility, sadness, tension, and fatigue, and significant reductions were observed in caring, relaxation, mental clarity, and vigor. In contrast, after listening to the designer music (music designed to have specific effects on the listener), significant increases in caring, relaxation, mental clarity, and vigor were measured; significant decreases were found in hostility, fatigue, sadness, and tension. The results for New Age and classical music were mixed. Feeling shifts among subjects were observed with all types of music. Designer music was most effective in increasing positive feelings and decreasing negative feelings. Results suggest that designer music may be useful in the treatment of tension, mental distraction, and negative moods.


Dynamic holography for optical interconnections. II. Routing holograms with predictable location and intensity of each diffraction order.
An analysis of dynamic phase-only holograms, described by fractional notation and recorded onto a pixelated spatial light modulator (SLM) in a reconfigurable optical beam-steering switch, is presented. The phase quantization and arrangement of the phase states and the SLM pixelation and dead-space effects are decoupled, expressed analytically, and simulated numerically. The phase analysis with a skip-rotate rule reveals the location and intensity of each diffraction order at the digital replay stage. The optical reconstruction of the holograms recorded onto SLM's with rectangular pixel apertures entails sinc-squared scaling, which further reduces the intensity of each diffraction order. With these two factors taken into account, the highest values of the nonuniform first-order diffraction efficiencies are expected to be 33%, 66%, and 77% for two-, four-, and and eight-level one-dimensional holograms with a 90% linear pixel fill factor. The variation of the first-order diffraction efficiency and the relative replay intensities were verified to within 1 dB by performing the optical reconstruction of binary phase-only holograms recorded onto a ferroelectric liquid crystal on a silicon SLM.


Atherogenic and anti-atherogenic factors in the human diet.
New atherosclerosis causative factors and preventive modalities have been identified. Atherogenic factors include lipid oxidation products, such as cholesterol oxidation products, malonaldehyde and other aldehydes; trans-fatty acids; some saturated fatty acids (lauric, myristic and possibly palmitic acids); and myristic acid plus cholesterol. Lipid oxidation products are well suited to induce arterial damage, based on their known cytotoxic effects; evidence also indicates the possibility of plaque promotion and stimulation of thrombogenesis. Anti-atherogenic factors include antioxidants, fish oils and other polyunsaturates (if protected from oxidation), fibre and trace minerals such as copper, manganese, selenium and zinc. Iron is unique, being considered as both a potential promoter of atherosclerosis (component of ferritin, conceivably inducing lipid oxidation) and a possible anti-atherogenic component (of antioxidant enzyme catalase). It is apparent that an entire new series of research challenges has been uncovered.


The outcome of B-cell receptor signaling in chronic lymphocytic leukemia: proliferation or anergy.
Biologists and clinicians agree that the B-cell receptor influences the behavior of chronic lymphocytic leukemia, and promising new drugs are aimed at receptor-associated kinases. Engagement of surface immunoglobulin by antigen is a key driver of malignant cells with outcome influenced by the nature of the cell, the level of stimulation and the microenvironment. Analysis of surface immunoglobulin-mediated signaling in the two major disease subsets defined by IGHV mutational status reveals bifurcation of responses toward proliferation or anergy. Mutated chronic lymphocytic leukemia, generally of relatively good prognosis, is mainly, but not exclusively, driven towards anergy in vivo. In contrast, unmutated chronic lymphocytic leukemia shows less evidence for anergy in vivo retaining more responsiveness to surface immunoglobulin M-mediated signaling, possibly explaining increased tumor progression. Expression and function of surface immunoglobulin M in unmutated chronic lymphocytic leukemia appear rather homogeneous, but mutated chronic lymphocytic leukemia exhibits a highly heterogeneous profile that may relate to further variable clinical behavior within this subset. Anergy should increase susceptibility to apoptosis but, in leukemic cells, this may be countered by overexpression of the B-cell lymphoma-2 survival protein. Maintained anergy spreads to chemokines and adhesion molecules, restraining homing and migration. However, anergy is not necessarily completely benign, being able to reverse and regenerate surface immunoglobulin M-mediated responses. A two-pronged attack on proliferative and anti-apoptotic pathways may succeed. Increased understanding of how chronic lymphocytic leukemia cells are driven to anergy or proliferation should reveal predictive biomarkers of progression and of likely response to kinase inhibitors, which could assist therapeutic decisions.


Cytotoxic efficacy of photodynamic therapy in osteosarcoma cells in vitro.
In recent years, there has been the difficulty in finding more effective therapies against cancer with less systemic side effects. Therefore Photodynamic Therapy is a novel approach for a more tumor selective treatment. Photodynamic Therapy (PDT) that makes use of a nontoxic photosensitizer (PS), which, upon activation with light of a specific wavelength in the presence of oxygen, generates oxygen radicals that elicit a cytotoxic response(1). Despite its approval almost twenty years ago by the FDA, PDT is nowadays only used to treat a limited number of cancer types (skin, bladder) and nononcological diseases (psoriasis, actinic keratosis)(2). The major advantage of the use of PDT is the ability to perform a local treatment, which prevents systemic side effects. Moreover, it allows the treatment of tumors at delicate sites (e.g. around nerves or blood vessels). Here, an intraoperative application of PDT is considered in osteosarcoma (OS), a tumor of the bone, to target primary tumor satellites left behind in tumor surrounding tissue after surgical tumor resection. The treatment aims at decreasing the number of recurrences and at reducing the risk for (postoperative) metastasis. In the present study, we present in vitro PDT procedures to establish the optimal PDT settings for effective treatment of widely used OS cell lines that are used to reproduce the human disease in well established intratibial OS mouse models. The uptake of the PS mTHPC was examined with a spectrophotometer and phototoxicity was provoked with laser light excitation of mTHPC at 652 nm to induce cell death assessed with a WST-1 assay and by the counting of surviving cells. The established techniques enable us to define the optimal PDT settings for future studies in animal models. They are an easy and quick tool for the evaluation of the efficacy of PDT in vitro before an application in vivo.


Time course of cerebrovascular autoregulation during extreme Trendelenburg position for robotic-assisted prostatic surgery.
Trendelenburg positioning in combination with pneumoperitoneum during robotic-assisted prostatic surgery possibly impairs cerebrovascular autoregulation. If cerebrovascular autoregulation is disturbed, arterial hypertension might induce cerebral hyperaemia and brain oedema, while low arterial blood pressure can induce cerebral ischaemia. The time course of cerebrovascular autoregulation was investigated during use of the Trendelenburg position and a pneumoperitoneum for robotic-assisted prostatic surgery using transcranial Doppler ultrasound. Cerebral blood flow velocity was correlated with arterial blood pressure and the autoregulation index (Mx) was calculated. In 23 male patients, Mx was assessed at baseline, after induction of general anaesthesia, during the Trendelenburg position (40-45°), and after repositioning. During the Trendelenburg position, Mx increased over time, indicating an impairment of cerebrovascular autoregulation. After repositioning, Mx recovered to baseline levels. It can be concluded that with longer durations of Trendelenburg position and pneumoperitoneum, cerebrovascular autoregulation deteriorates, and, therefore, blood pressure management should be adapted to avoid cerebral oedema and the duration of Trendelenburg position should be as short as possible.


Blau syndrome, the prototypic auto-inflammatory granulomatous disease.
Blau syndrome is a monogenic disease resulting from mutations in the pattern recognition receptor NOD2, and is phenotypically characterized by the triad of granulomatous polyarthritis, dermatitis and uveitis. This paper reviews briefly the classical clinical features of the disease, as well as more recently described extra-triad symptoms. From an ongoing prospective multicenter study, we provide new data on the natural history of Blau syndrome, focusing on functional status and visual outcome. We also present an update of the range of different NOD2 mutations found in Blau syndrome as well as recent data on morphologic and immunohistochemical characteristics of the Blau granuloma. Finally, emerging insights into pathogenic mechanisms including activation of NOD2 signal transduction, and potential biomarkers of disease activity are discussed.


Biological embedding of child maltreatment: A systematic review of biomarkers and resilience in children and youth.
Child maltreatment (CM) is a widespread problem associated with poor mental and physical health outcomes. The underlying mechanisms of this link are not always well understood, however certain biological changes observed in maltreated individuals may play a role in connecting experience and outcome. This review specifically focuses on 2 markers of biological embedding, DNA methylation (DNAm) and telomere length (TL) in maltreated children and youth. As biomarker changes are not uniform among maltreated children, we additionally discuss biological and environmental resilience factors that may contribute to variability. We conducted a systematic review of Medline, Embase and PsycINFO databases for studies examining DNAm and/or TL in maltreated children and youth. Methodological quality of the included studies was assessed using the Scottish Intercollegiate Guidelines Network (SIGN) checklists for cohort studies and randomized control trials. Data extraction focused on various factors including population and CM (type, chronicity, severity, and duration) characteristics. The initial search returned 1,688 nonduplicate results, with 417 full text articles reviewed. Twenty-six articles from 16 studies were ultimately included of which 8 examined telomere length and 18 examined DNA methylation. While some heterogeneity of findings was found, evidence supports differential changes in both biomarkers associated with CM. This review enhances understanding of the constellation of biological changes related to CM and consideration of the important role of resilience factors in mitigating risk. Elucidating these factors may highlight targets for future study and intervention development. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Compensation for pixel misregistration in volume holographic data storage.
We describe what we believe to be a novel postprocessing algorithm for compensating for misregistrations between a detector array and the coherent image of a pixelated two-dimensional data page. A lookup table of baseline local offsets is combined with the dynamically measured global offset of the received data page, producing an estimate of the total lateral shift of each small block of pixels. A serial algorithm then reallocates the appropriate portion of the signal detected by each pixel to its neighbors, accounting for both the linear and the quadratic contributions introduced by coherent illumination of square-law detectors. This procedure can relax the tight constraints on page registration, optical distortion, and material shrinkage that currently hamper page-oriented holographic storage systems. Experimental results from a pixel-matched 1-Mpixel volume holographic system are presented, showing an increase in position tolerance (for a raw bit-error rate <10(-3)) from +/-16% to +/-40% of the pixel pitch.


Engaging Patients, Health Care Professionals, and Community Members to Improve Preoperative Decision Making for Older Adults Facing High-Risk Surgery.
Older patients are at greater risk for postoperative complications, yet they are less likely than younger patients to ask questions about surgery. To design an intervention to improve preoperative decision making and manage postoperative expectations. A Patient and Family Advisory Council (PFAC) was created to help identify preoperative decisional needs. The PFAC included 4 men and women who had previous experience with high-risk surgery as older patients or their family members; the PFAC met monthly at a local library from May 2014 to April 2015 to examine findings from a prior qualitative study and to integrate themes with PFAC members' experiences. Patient observations included 91 recorded conversations between patients and surgeons and 61 patient interviews before and after surgery. The PFAC members and other stakeholders evaluated 118 publicly available questions and selected 12 corresponding to identified needs to generate a question prompt list (QPL). Three focus groups, including 31 community members from diverse backgrounds, were conducted at community centers in Madison and Milwaukee, Wisconsin, to refine the QPL. A clinical pilot with 42 patients considering surgery was conducted in one outpatient surgical clinic in Madison. Generation of a QPL to address patients' preoperative informational and decisional needs. Through exploration of qualitative data, the PFAC noted 3 critical problems. Patients and family members believed surgery had to be done, were surprised that postoperative recovery was difficult, and lacked knowledge about the perioperative use of advance directives. The PFAC identified a need for more information and decisional support during preoperative conversations that included clarification of treatment options, setting postoperative expectations, and advance care planning. The following 3 question prompt categories arose: "Should I have surgery?" "What should I expect if everything goes well?" and "What happens if things go wrong?" The final list included 11 questions within these domains, was understandable in English and Spanish, and was acceptable to patients in the clinic. Through direct engagement of stakeholders, a QPL was created to address core decisional and informational needs of surgical patients. Future testing will evaluate whether this list can be used to improve patient engagement and reduce postoperative regret and conflict about postoperative treatments.


Geographic Variation of the Incidence Rate of Lower Limb Amputation in Australia from 2007-12.
In Australia, little is known about how the incidence rate (IR) of lower limb amputation (LLA) varies across the country. While studies in other economically developed countries have shown considerable geographic variation in the IR-LLA, mostly these have not considered whether the effect of common risk factors are the same across regions. Mapping variation of the IR-LLA, and the effect of common risk factors, is an important first step to focus research into areas of greatest need and support the development of regional specific hypotheses for in-depth examination. The aim of this study was to describe the geographic variation in the IR-LLA across Australia and understand whether the effect of common risk factors was the same across regions. Using hospital episode data from the Australian National Hospital Morbidity database and Australian Bureau of Statistics, the all-cause crude and age-standardised IR-LLA in males and females were calculated for the nation and each state and territory. Generalised Linear Models were developed to understand which factors influenced geographic variation in the crude IR-LLA. While the crude and age-standardised IR-LLA in males and females were similar in most states and territories, they were higher in the Northern Territory. The effect of older age, being male and the presence of type 2 diabetes was associated with an increase of IR-LLA in most states and territories. In the Northern Territory, the younger age at amputation confounded the effect of sex and type 2 diabetes. There are likely to be many factors not included in this investigation, such as Indigenous status, that may explain part of the variation in the IR-LLA not captured in our models. Further research is needed to identify regional- and population- specific factors that could be modified to reduce the IR-LLA in all states and territories of Australia.


A dynamic model of the jump-to phenomenon during AFM analysis.
The measurement of the physical properties of surfaces on the nanoscale is a long-standing problem, and the atomic force microscope (AFM) has enabled the investigation of surface energies and mechanical properties over a range of length scales. The ability to measure these properties for softer materials presents a challenge when interpreting data obtained from such measurements, in particular because of the dynamics of the compliant AFM microcantilever. This work attempts to better understand the interaction between an AFM tip and samples of varying elastic modulus, in the presence of attractive van der Waals forces. A theoretical model is presented in which the dynamics of the approach of an atomic force microscope cantilever tip toward a surface, prior to and during the van der Waals-induced jump-to phenomenon, are included. The cantilever mechanics incorporates the motion of the air through which the cantilever moves, the acceleration, inertia, and torque of the cantilever, and the squeezing of the fluid between the cantilever tip and the surface, leading to elastohydrodynamic lubrication and deformation of the substrate. Simulations of the cantilever approach are compared to measurements performed using an atomic force microscope, and the effect of cantilever drive velocity is investigated. Cantilevers presenting (1) spherical colloid probe tips and (2) pyramidal tips are employed, and substrates exhibiting Young's moduli of 3 MPa, 500 MPa, and 75 GPa are measured. The analysis presented could be extended to enhance understanding of dynamic phenomena in micro/nanoelectromechanical systems such as resonators and microrheometers, particularly those which contain soft materials and also where surface interactions are important.


Myelin repair and functional recovery mediated by neural cell transplantation in a mouse model of multiple sclerosis.
Cellular therapies are becoming a major focus for the treatment of demyelinating diseases such as multiple sclerosis (MS), therefore it is important to identify the most effective cell types that promote myelin repair. Several components contribute to the relative benefits of specific cell types including the overall efficacy of the cell therapy, the reproducibility of treatment, the mechanisms of action of distinct cell types and the ease of isolation and generation of therapeutic populations. A range of distinct cell populations promote functional recovery in animal models of MS including neural stem cells and mesenchymal stem cells derived from different tissues. Each of these cell populations has advantages and disadvantages and likely works through distinct mechanisms. The relevance of such mechanisms to myelin repair in the adult central nervous system is unclear since the therapeutic cells are generally derived from developing animals. Here we describe the isolation and characterization of a population of neural cells from the adult spinal cord that are characterized by the expression of the cell surface glycoprotein NG2. In functional studies, injection of adult NG2(+) cells into mice with ongoing MOG35-55-induced experimental autoimmune encephalomyelitis (EAE) enhanced remyelination in the CNS while the number of CD3(+) T cells in areas of spinal cord demyelination was reduced approximately three-fold. In vivo studies indicated that in EAE, NG2(+) cells stimulated endogenous repair while in vitro they responded to signals in areas of induced inflammation by differentiating into oligodendrocytes. These results suggested that adult NG2(+) cells represent a useful cell population for promoting neural repair in a variety of different conditions including demyelinating diseases such as MS.


Effect of insulinlike growth factor I on DNA and protein synthesis in cultured rat calvaria.
Insulinlike growth Factor I (IGF I), a growth hormone-dependent peptide or somatomedin, was studied for its effects on bone formation by examining the synthesis of DNA, collagen, and noncollagen protein in cultures of 21-d fetal rat calvaria. IGF I caused a dose-dependent stimulation of the incorporation of [3H]thymidine into DNA at concentrations of 0.1--100 nM; the effect appeared after 6 h, was maximal at 12 h, and was sustained for 96 h. IGF I also increased the bone DNA content, IGF I at 0.1--3 nM had a small stimulatory effect on the incorporation of [3H]proline into collagenase-digestible protein (CDP) whereas 30 nM IGF I caused a two- to threefold increment and had a maximal effect. A smaller effect on the labeling of noncollagen protein (NCP) was also observed. The effect of CDP and NCP appeared and was maximal after 12 h and was sustained for 96 h. IGF I increased the total collagen content of bones. The IGF I stimulatory effect on the incorporation of [3H]thymidine was seen in both the periosteum and periosteum-free calvarium, whereas that on the labeling of CDP was seen only in the central, osteoblastic-rich, non-periosteal bone. Histological sections showed a 10-fold increase in the mitotic index after Colcemid arrest in IGF I-treated bones, the mitoses were equally distributed in the periosteum and central portions of the calvarium. Insulin had a stimulatory effect on the incorporation of [3H]proline into CDP and NCP and 1 nM--1 microM similar to the effect of IGF I. In contrast, high insulin concentrations (0.1 and 1 microM) were required to increase the incorporation of [3H]thymidine, and insulin did not affect DNA content. Cortisol decreased the stimulatory effect of IGF I on DNA labeling but greatly enhanced the stimulatory effect of IGF I on the incorporation of [3H]proline into CDP. Triiodothyronine and parathyroid hormone increased the incorporation of [3H]thymidine and were additive to IGF I. Triiodothyronine did not affect the labeling of CDP, but parathyroid hormone inhibited it and opposed the effect of IGF I. These studies indicate that IGF I stimulates bone DNA, collagen, and NCP synthesis in vitro. IGF I and insulin have similar effects on bone collagen synthesis but IGF I stimulates the synthesis of DNA at physiological concentrations, and insulin does not.


Prefrontal-Periaqueductal Gray-Projecting Neurons Mediate Context Fear Discrimination.
Survival critically depends on selecting appropriate defensive or exploratory behaviors and is strongly influenced by the surrounding environment. Contextual discrimination is a fundamental process that is thought to depend on the prefrontal cortex to integrate sensory information from the environment and regulate adaptive responses to threat during uncertainty. However, the precise prefrontal circuits necessary for discriminating a previously threatening context from a neutral context remain unknown. Using a combination of single-unit recordings and optogenetic manipulations, we identified a neuronal subpopulation in the dorsal medial prefrontal cortex (dmPFC) that projects to the lateral and ventrolateral periaqueductal gray (l/vlPAG) and is selectively activated during contextual fear discrimination. Moreover, optogenetic activation and inhibition of this neuronal population promoted contextual fear discrimination and generalization, respectively. Our results identify a subpopulation of dmPFC-l/vlPAG-projecting neurons that control switching between different emotional states during contextual discrimination.


Development of PET/SPECT ligands for the serotonin transporter.
There is a great need for PET and SPECT ligands with high affinity and selectivity for the serotonin uptake site. These imaging agents would be useful in screening human populations at risk (e.g., individuals exposed to neurotoxic amphetamines such as MDMA and fenfluramine). Moreover, these radioligands would allow the study of serotonergic function in the normal living human brain, and they also would be useful in the examination of altered serotonergic neurotransmission in diseases such as depression and obsessive-compulsive and other neuropsychiatric disorders. Over the past several years, a number of radioligands have been studied in several laboratories for their in vivo binding to 5-HT uptake sites. Although [3H]paroxetine showed promising binding characteristics, conversion of authentic paroxetine into a PET or SPECT tracer turned out to be difficult and has not been achieved yet. Analogs of paroxetine displayed considerable loss of binding affinity and were, therefore, not useful for imaging purposes. For [11C]fluoxetine, [11C]citalopram, and cis-[11C]DDPI, target-to-nontarget (hypothalamus-to-cerebellar) ratios remained less than 2.0:1 over a 90-min period after injection. The most promising PET agents identified today are [11C]RTI-55 and [11C]McN-5652-X. [11C]RTI-55 labels both 5-HT and DA uptake sites. [11C]McN-5652-X is highly selective for 5-HT uptake sites, and its distribution is consistent with the neuroanatomical distribution of the 5-HT uptake site. Because [11C]McN-5652-Z is a racemic mixture of two stereoisomers, of which the (+) isomer (McN-5652-X) binds to the 5-HT uptake site in vivo and the (-) isomer (McN-5652-W) does not, the possibility exists that regional-specific binding can be determined by subtracting nonspecific binding of the (-) isomer from total radioactivity counts obtained with the (+) isomer. [11C]McN-5652-X is the best PET radioligand for the 5-HT uptake site described thus far. This tracer warrants further testing in nonhuman primates. Efforts are underway to obtain an investigational new drug application for use of the tracer in humans. Promising candidates as SPECT imaging agents for the 5-HT uptake site are [123I]RTI-55 and [123I]-iodo-6-nitroquipazine. Both agents are under intense investigation in different laboratories in the United States.


Genital tract infections, the vaginal microbiome and gestational age at birth among pregnant women in South Africa: a cohort study protocol.
Preterm birth complications are the most common cause of death in children under 5 years. The presence of multiple microorganisms and genital tract inflammation could be the common mechanism driving early onset of labour. South Africa has high levels of preterm birth, genital tract infections and HIV infection among pregnant women. We plan to investigate associations between the presence of multiple lower genital tract microorganisms in pregnancy and gestational age at birth. This cohort study enrols around 600 pregnant women at one public healthcare facility in East London, South Africa. Eligible women are ≥18 years and at <27 weeks of gestation, confirmed by ultrasound. At enrolment and 30-34 weeks of pregnancy, participants receive on-site tests for Chlamydia trachomatis and Neisseria gonorrhoeae, with treatment if test results are positive. At these visits, additional vaginal specimens are taken for: PCR detection and quantification of Trichomonas vaginalis, Candida spp., Mycoplasma genitalium, M. hominis, Ureaplasma urealyticum and U. parvum; microscopy and Nugent scoring; and for 16S ribosomal RNA gene sequencing and quantification. Pregnancy outcomes are collected from a postnatal visit and birth registers. The primary outcome is gestational age at birth. Statistical analyses will explore associations between specific microorganisms and gestational age at birth. To explore the association with the quantity of microorganisms, we will construct an index of microorganism load and use mixed-effects regression models and classification and regression tree analysis to examine which combinations of microorganisms contribute to earlier gestational age at birth. This protocol has approvals from the University of Cape Town Research Ethics Committee and the Canton of Bern Ethics Committee. Results from this study will be uploaded to preprint servers, submitted to open access peer-reviewed journals and presented at regional and international conferences. NCT06131749; Pre-results.


A global trait-based approach to estimate leaf nitrogen functional allocation from observations.
Nitrogen is one of the most important nutrients for plant growth and a major constituent of proteins that regulate photosynthetic and respiratory processes. However, a comprehensive global analysis of nitrogen allocation in leaves for major processes with respect to different plant functional types (PFTs) is currently lacking. This study integrated observations from global databases with photosynthesis and respiration models to determine plant-functional-type-specific allocation patterns of leaf nitrogen for photosynthesis (Rubisco, electron transport, light absorption) and respiration (growth and maintenance), and by difference from observed total leaf nitrogen, an unexplained "residual" nitrogen pool. Based on our analysis, crops partition the largest fraction of nitrogen to photosynthesis (57%) and respiration (5%) followed by herbaceous plants (44% and 4%). Tropical broadleaf evergreen trees partition the least to photosynthesis (25%) and respiration (2%) followed by needle-leaved evergreen trees (28% and 3%). In trees (especially needle-leaved evergreen and tropical broadleaf evergreen trees) a large fraction (70% and 73%, respectively) of nitrogen was not explained by photosynthetic or respiratory functions. Compared to crops and herbaceous plants, this large residual pool is hypothesized to emerge from larger investments in cell wall proteins, lipids, amino acids, nucleic acid, CO2 fixation proteins (other than Rubisco), secondary compounds, and other proteins. Our estimates are different from previous studies due to differences in methodology and assumptions used in deriving nitrogen allocation estimates. Unlike previous studies, we integrate and infer nitrogen allocation estimates across multiple PFTs, and report substantial differences in nitrogen allocation across different PFTs. The resulting pattern of nitrogen allocation provides insights on mechanisms that operate at a cellular scale within leaves, and can be integrated with ecosystem models to derive emergent properties of ecosystem productivity at local, regional, and global scales.


Prevalence of hepatitis A, hepatitis B, and HIV among hepatitis C-seropositive state hospital patients: results from Oregon State Hospital.
Multiple studies have shown that individuals with severe mental illness are at increased risk for acquiring infection from human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). Moreover, patients with chronic HCV infection are at risk for fulminant hepatitis from acquired infection with hepatitis A virus (HAV) or HBV, but there are limited data on the prevalence of HIV, HAV, and HBV in chronically hospitalized U.S. psychiatric patients without mental retardation who are HCV-seropositive. To address this issue, a comprehensive screening program was commenced at Oregon State Hospital (Salem, Ore.) beginning in 1999. The computerized records of all non-geriatric adult inpatients at Oregon State Hospital on April 23, 2001, were reviewed to assess physician compliance with screening and the prevalence of infection with HIV, HAV, HBV, and HCV. Among the 535 patient records reviewed, 94.8% of patients were screened for HCV, of whom 20.3% were seropositive. Among HCV-seropositive patients, only 1.9% were not screened for HAV and HBV, but 23.3% were not tested for HIV. In the HCV-seropositive group, 35.9% were HAV-positive, 49.5% HBV-positive, and 2.6% HIV-positive. Chronic psychiatric inpatients have high HCV prevalence rates. Hepatitis C-seropositive individuals may be at risk for complications unless vaccinated for HAV and HBV.


InterMitoBase: an annotated database and analysis platform of protein-protein interactions for human mitochondria.
The mitochondrion is an essential organelle which plays important roles in diverse biological processes, such as metabolism, apoptosis, signal transduction and cell cycle. Characterizing protein-protein interactions (PPIs) that execute mitochondrial functions is fundamental in understanding the mechanisms underlying biological functions and diseases associated with mitochondria. Investigations examining mitochondria are expanding to the system level because of the accumulation of mitochondrial proteomes and human interactome. Consequently, the development of a database that provides the entire protein interaction map of the human mitochondrion is urgently required. InterMitoBase provides a comprehensive interactome of human mitochondria. It contains the PPIs in biological pathways mediated by mitochondrial proteins, the PPIs between mitochondrial proteins and non-mitochondrial proteins as well as the PPIs between mitochondrial proteins. The current version of InterMitoBase covers 5,883 non-redundant PPIs of 2,813 proteins integrated from a wide range of resources including PubMed, KEGG, BioGRID, HPRD, DIP and IntAct. Comprehensive curations have been made on the interactions derived from PubMed. All the interactions in InterMitoBase are annotated according to the information collected from their original sources, GenBank and GO. Additionally, InterMitoBase features a user-friendly graphic visualization platform to present functional and topological analysis of PPI networks identified. This should aid researchers in the study of underlying biological properties. InterMitoBase is designed as an integrated PPI database which provides the most up-to-date PPI information for human mitochondria. It also works as a platform by integrating several on-line tools for the PPI analysis. As an analysis platform and as a PPI database, InterMitoBase will be an important database for the study of mitochondria biochemistry, and should be particularly helpful in comprehensive analyses of complex biological mechanisms underlying mitochondrial functions.


Visualization of retinal vascular structure and perfusion with a nonconfocal adaptive optics scanning light ophthalmoscope.
Imaging of the retinal vascular structure and perfusion was explored by confocal illumination and nonconfocal detection in an adaptive optics scanning light ophthalmoscope (AOSLO), as an extension of the work by Chui et al. [Biomed. Opt. Express 3, 2537 (2012)]. Five different detection schemes were evaluated at multiple retinal locations: circular mask, annular mask, circular mask with filament, knife-edge, and split-detector. Given the superior image contrast in the reflectance and perfusion maps, the split-detection method was further tested using pupil apodization, polarized detection, and four different wavelengths. None of these variations provided noticeable contrast improvement. The noninvasive visualization of capillary flow and structure provided by AOSLO split-detection shows great promise for studying ocular and systemic conditions that affect the retinal vasculature.


The effect of chelating agents on iron mobilization in Chang cell cultures.
The investigation of chelating agents with potential therapeutic value in patients with transfusional iron overload has been facilitated by the use of Chang cell cultures. These cells have been incubated with [59Fe]transferrin for 22 hr, following which most of the intracellular radioiron is found in the cytosol, distributed between a ferritin and a nonferritin form. Iron release from the cells depends on transferrin saturation in the medium, but when transferrin is 100% saturated, which normally does not allow iron release, desferrioxamine, 2,3-dihydroxybenzoic acid, rhodotorulic acid, cholythydroxamic acid, and tropolone all promote the mobilization of ferritin iron and its release from cells. They are effective to an approximately equal degree. The incubation of [59Fe]transferrin with tropolone in vitro at a molar ratio of 1:500 results in the transfer of most of the labeled iron to the chelator, reflecting the exceptionally high binding constant of this compound. How far these phenomena relate to therapeutic potentially remains to be seen.


Antidepressants and suicide risk: how did specific information in FDA safety warnings affect treatment patterns?
From June 2003 through October 2004, the U.S. Food and Drug Administration (FDA) released five safety warnings related to antidepressant use and the increased risk of suicidality for children. Although researchers have documented a decline in antidepressant use among children over this period, less is known about whether specific safety information conveyed in individual warnings was reflected in treatment patterns. Thomson Reuters MarketScan claims data (2001-2005) for a national sample of privately insured children were used to construct treatment episodes (N=22,689). For each new episode of major depressive disorder, it was determined whether treatment followed specific recommendations included in warnings released by the FDA. Treatment recommendations pertained to the use of the antidepressants paroxetine and fluoxetine and to patient monitoring. Treatment patterns were expected to change as the risk information conveyed by the FDA changed over time. The timing of FDA recommendations was associated with trends in the use of paroxetine and fluoxetine by children with major depressive disorder who were initiating antidepressant treatment. However, no evidence of increases in outpatient visits (indicative of monitoring) among depressed children initiating antidepressant use was found. Release of specific risk and benefit information by the FDA was associated with changes in prescribing but not in outpatient follow-up. These results suggest that the FDA plays an important role in communicating information to the public and providers. Yet, although public health safety warnings were associated with changes in some practice patterns, not all recommendations conveyed in warnings were followed.


Association of Plasma Neurofilament Light with Postoperative Delirium.
To examine the association of the plasma neuroaxonal injury markers neurofilament light (NfL), total tau, glial fibrillary acid protein, and ubiquitin carboxyl-terminal hydrolase L1 with delirium, delirium severity, and cognitive performance. Delirium case-no delirium control (n = 108) pairs were matched by age, sex, surgery type, cognition, and vascular comorbidities. Biomarkers were measured in plasma collected preoperatively (PREOP), and 2 days (POD2) and 30 days postoperatively (PO1MO) using Simoa technology (Quanterix, Lexington, MA). The Confusion Assessment Method (CAM) and CAM-S (Severity) were used to measure delirium and delirium severity, respectively. Cognitive function was measured with General Cognitive Performance (GCP) scores. Delirium cases had higher NfL on POD2 and PO1MO (median matched pair difference = 16.2pg/ml and 13.6pg/ml, respectively; p < 0.05). Patients with PREOP and POD2 NfL in the highest quartile (Q4) had increased risk for incident delirium (adjusted odds ratio [OR] = 3.7 [95% confidence interval (CI) = 1.1-12.6] and 4.6 [95% CI = 1.2-18.2], respectively) and experienced more severe delirium, with sum CAM-S scores 7.8 points (95% CI = 1.6-14.0) and 9.3 points higher (95% CI = 3.2-15.5). At PO1MO, delirium cases had continued high NfL (adjusted OR = 9.7, 95% CI = 2.3-41.4), and those with Q4 NfL values showed a -2.3 point decline in GCP score (-2.3 points, 95% CI = -4.7 to -0.9). Patients with the highest PREOP or POD2 NfL levels were more likely to develop delirium. Elevated NfL at PO1MO was associated with delirium and greater cognitive decline. These findings suggest NfL may be useful as a predictive biomarker for delirium risk and long-term cognitive decline, and once confirmed would provide pathophysiological evidence for neuroaxonal injury following delirium. ANN NEUROL 2020;88:984-994.


Open-Source Automated Parahydrogen Hyperpolarizer for Molecular Imaging Using (13)C Metabolic Contrast Agents.
An open-source hyperpolarizer producing (13)C hyperpolarized contrast agents using parahydrogen induced polarization (PHIP) for biomedical and other applications is presented. This PHIP hyperpolarizer utilizes an Arduino microcontroller in conjunction with a readily modified graphical user interface written in the open-source processing software environment to completely control the PHIP hyperpolarization process including remotely triggering an NMR spectrometer for efficient production of payloads of hyperpolarized contrast agent and in situ quality assurance of the produced hyperpolarization. Key advantages of this hyperpolarizer include: (i) use of open-source software and hardware seamlessly allowing for replication and further improvement as well as readily customizable integration with other NMR spectrometers or MRI scanners (i.e., this is a multiplatform design), (ii) relatively low cost and robustness, and (iii) in situ detection capability and complete automation. The device performance is demonstrated by production of a dose (∼2-3 mL) of hyperpolarized (13)C-succinate with %P13C ∼ 28% and 30 mM concentration and (13)C-phospholactate at %P13C ∼ 15% and 25 mM concentration in aqueous medium. These contrast agents are used for ultrafast molecular imaging and spectroscopy at 4.7 and 0.0475 T. In particular, the conversion of hyperpolarized (13)C-phospholactate to (13)C-lactate in vivo is used here to demonstrate the feasibility of ultrafast multislice (13)C MRI after tail vein injection of hyperpolarized (13)C-phospholactate in mice.


The Sertoli-spermatid junctional complex adhesion strength is affected in vitro by adjudin.
The actin-based cell-cell adherens junction (AJ) between the Sertoli cell and the germ cell in the mammalian testis is important not only in mechanical adhesion of the cells, but in the morphogenesis and differentiation of the germ cells. The Sertoli ectoplasmic specialization (ES), a specialized type of AJ, is associated with Sertoli-spermatid binding and is important in cell-cell adhesion in the seminiferous epithelium. Abnormal or absent Sertoli ESs have been associated with step-8 spermatid sloughing and oligospermia in conditions associated with reduced fertility potential. The reproductive hormones, follicle stimulating hormone (FSH), and testosterone (T) have also been shown to play a role in the regulation of binding of spermatids at the Sertoli-spermatid junctional complex (STJC). Adjudin [1-(2,4-dichlorobenzyl)-1H-indazole-3-carbohydrazide] is a potential male contraceptive and is thought to exhibit its contraceptive effects by interrupting the STJC. It has been shown that this compound induces reversible germ cell loss from the seminiferous epithelium, particularly elongating/elongate/round spermatids and spermatocytes. Using a micropipette pressure transducing system (MPTS) to measure the force needed to detach step-8 spermatids from Sertoli cells, this study examined the strength of the STJC in Sertoli-spermatid cocultures in the presence of Adjudin (1 ng/mL, 50 ng/mL, 125 ng/mL, or 500 ng/mL in EtOH) and hormones [FSH (0.1 micro g/mL, NIDDK-oFSH-20, AFP7028D, 175 x NIH-FSH-S1), T (100 nM)] to optimize in vitro binding. The average forces required to detach the spermatids from the underlying Sertoli cells in the presence of 1 ng/mL, 50 ng/mL, 125 ng/mL, and 500 ng/mL Adjudin were 18.2 x 10(-10) pN, 14.3 x 10(-10) pN, 7.74 x 10(-10) pN, and 6.51 x 10(-10) pN, respectively. The average force required to detach step-8 spermatids in the presence of vehicle only (control) was 19.0 x 10(-10) pN. A significant difference for Adjudin concentrations at or above 125 ng/mL was determined by one-way ANOVA (P < .05). These data confirm that Adjudin is effective in reducing the strength of the STJC, identifying Adjudin as a potential contraceptive agent in the male by inducing spermatid sloughing and therefore oligospermia.


Structural diversity of Alzheimer's disease amyloid-β dimers and their role in oligomerization and fibril formation.
Alzheimer's disease (AD) is associated with the formation of toxic amyloid-β (Aβ)42 oligomers, and recent evidence supports a role for Aβ dimers as building blocks for oligomers. Molecular dynamics simulation studies have identified clans for the dominant conformations of Aβ42 forming dimers; however, it is unclear if a larger spectrum of dimers is involved and which set(s) of dimers might evolve to oligomers verse fibrils. Therefore, for this study we generated multiple structural conformations of Aβ42, using explicit all-atom molecular dynamics, and then clustering the different structures based on key conformational similarities. Those matching a selection threshold were then used to model a process of oligomerization. Remarkably, we showed a greater diversity in Aβ dimers than previously described. Depending on the clan family, different types of Aβ dimers were obtained. While some had the tendency to evolve into oligomeric rings, others formed fibrils of diverse characteristics. Then we selected the dimers that would evolve to membranephilic annular oligomers. Nearly one third of the 28 evaluated annular oligomers had the dimer interfaces between the neighboring Aβ42 monomers with possible salt bridges between the residue K28 from one side and either residue E22 or D23 on the other. Based on these results, key amino acids were identified for point mutations that either enhanced or suppressed the formation and toxicity of oligomer rings. Our studies suggest a greater diversity of Aβ dimers. Understanding the structure of Aβ dimers might be important for the rationale design of small molecules that block formation of toxic oligomers.


Modified CDKN2B (p15) and CDKN2A (p16) DNA methylation profiles in urban pesticide applicators.
Gene-specific changes in DNA methylation by pesticides in occupationally exposed populations have not been studied extensively. Of particular concern are changes in the methylation profile of tumor-suppressor, such as CDKN2B and CDKN2A, genes involved in oncogenesis. The aim of this study was to evaluate the methylation profiles of CDKN2B and CDKN2A genes in urban pesticide applicators and their relationship with occupational exposure to pesticides. A cross-sectional study was conducted in 186 urban pesticide applicators (categorized as high or moderate exposures) and 102 participants without documented occupational exposures to pesticides. Acute and chronic pesticide exposures were evaluated by direct measurement of urinary dialkylphosphates, organophosphate metabolites, and a structured questionnaire, respectively. Anthropometric characteristics, diet, clinical histories, and other variables were estimated through a validated self-reported survey. DNA methylation was determined by pyrosequencing of bisulfite-treated DNA. Decreased DNA methylation of the CDKN2B gene was observed in pesticide-exposed groups compared to the non-exposed group. In addition, increased methylation of the CDKN2A promoter was observed in the moderate-exposure group compared to the non-exposed group. Bivariate analysis showed an association between CDKN2B methylation and pesticide exposure, general characteristics, smoking status, and micronutrients, while changes in CDKN2A methylation were associated with pesticide exposure, sex, educational level, body mass index, smoking status, supplement intake, clinical parameters, and caffeine consumption. These data suggest that pesticide exposure modifies the methylation pattern of CDKN2B and CDKN2A genes and raise important questions about the role that these changes may play in the regulation of cell cycle activities, senescence, and aging.


Retrospective Parameter Estimation and Forecast of Respiratory Syncytial Virus in the United States.
Recent studies have shown that systems combining mathematical modeling and Bayesian inference methods can be used to generate real-time forecasts of future infectious disease incidence. Here we develop such a system to study and forecast respiratory syncytial virus (RSV). RSV is the most common cause of acute lower respiratory infection and bronchiolitis. Advanced warning of the epidemic timing and volume of RSV patient surges has the potential to reduce well-documented delays of treatment in emergency departments. We use a susceptible-infectious-recovered (SIR) model in conjunction with an ensemble adjustment Kalman filter (EAKF) and ten years of regional U.S. specimen data provided by the Centers for Disease Control and Prevention. The data and EAKF are used to optimize the SIR model and i) estimate critical epidemiological parameters over the course of each outbreak and ii) generate retrospective forecasts. The basic reproductive number, R0, is estimated at 3.0 (standard deviation 0.6) across all seasons and locations. The peak magnitude of RSV outbreaks is forecast with nearly 70% accuracy (i.e. nearly 70% of forecasts within 25% of the actual peak), four weeks before the predicted peak. This work represents a first step in the development of a real-time RSV prediction system.


Thiocyanate concentration in saliva of cystic fibrosis patients.
Thiocyanates (SCN-) are ubiquitous in nature. There are indispensable part of host defense system that act as a substrate for lactoperoxidase (LPO). In our study we present initial data on SCN- concentration in saliva of CF patients in comparison to healthy non-smokers and healthy smokers. 5 ml of saliva was collected from each subject to a sterile tube and thiocyanate concentration was measured in each sample. The results of the measurements are presented on Fig. 1. Mean concentration of SCN- in saliva of CF patients was 0.031 +/- 0.0052 g/l, in healthy non-smokers 0.039 +/- 0.0048 g/l and in healthy smokers 0.048 +/- 0.0161 g/l. The differences between each group were statistically significant. Studies on larger group of patients and probably on different material (BALF or induced sputum) should present interesting data complementing the in vitro studies.


Are hemianopic monkeys and a human hemianope aware of visual events in the blind field?
In three hemianopic monkeys and one normal monkey who subsequently became hemianopic and in one human hemianope we measured reaction times to touch the remembered position of a brief visual target presented in the normal hemifield or in the blind hemifield, or on the blank trials where no visual target occurred and the correct response was to touch a separate and permanently outlined part of the display. This is the same procedure as first used to demonstrate blindsight in these hemianopic monkeys. In the present experiment physically salient high-contrast (0.95) grating stimuli, which we have previously shown are easily detected and localized in the blind field, were used, and the monkeys' reaction times were also measured. With rare exceptions the monkeys indicated that the visual targets in the blind field were blanks, but their otherwise identical motor responses to targets and blanks had significantly different latencies, which were longer for real targets than for blanks. The results indicate that when the monkeys detect that the stimulus has occurred but do not perceive it as a light, or are just guessing, reaction times are longer. Even when the target in the blind field was moving, it was categorized as a blank. In the human hemianope both high- and low-contrast stimuli were used, and the subject had to indicate whether he had been 'aware' or 'unaware' of the target, after making the reaching response. The latencies when he was correct and aware were significantly shorter than when he was unaware and 'just guessing'. However, he was also significantly faster to respond correctly to the blind-field target when he was unaware and correct than when he was unaware and incorrect, a difference reflected in his percentage correct scores even when totally unaware. Collectively, the results support the notion that the hemianopic monkeys, like the human hemianope, know that something has happened in the blind field as long as the stimuli are physically salient even though the stimuli are categorized as blanks, presumably because, like the human hemianope, there was no phenomenal visual percept.


Toll-like receptor 4 prevents AOM/DSS-induced colitis-associated colorectal cancer in Bacteroides fragilis gnotobiotic mice.
Bacteroides fragilis (BF) plays a critical role in developing and maintaining the mammalian immune system. We previously found that BF colonization could prevent inflammation and tumor formation in a germ-free (GF) colitis-associated colorectal cancer (CAC) mouse model. The role of Toll-like receptor 4 (TLR4) in CAC development has not been clearly elucidated in BF mono-colonized gnotobiotic mice. The wild-type (WT) and TLR4 knockout (T4K) germ-free mice were raised with or without BF colonization for 28 days (GF/WT, GF/T4K, BF/WT, and BF/T4K) and then CAC was induced under azoxymethane (AOM)/dextran sulfate sodium (DSS) administration. The results showed that tumor formation and tumor incidence were significantly inhibited in the BF/WT group compared to those observed in the GF/WT group. However, the tumor prevention effect was not observed in the BF/T4K group unlike in the BF/WT group. Moreover, the CAC histological severity of the BF/WT group was ameliorated, but more severe lesions were found in the GF/WT, GF/T4K, and BF/T4K groups. Immunohistochemistry showed decreased cell proliferation (PCNA, β-catenin) and inflammatory markers (iNOS) in the BF/WT group compared to those in the BF/T4K group. Taken together, BF mono-colonization of GF mice might prevent CAC via the TLR4 signal pathway.


Manifestations of integrated public health policy in Dutch municipalities.
Integrated public health policy (IPHP) aims at integrating health considerations into policies of other sectors. Since the limited empirical evidence available may hamper its further development, we systematically analysed empirical manifestations of IPHP, by placing policy strategies along a continuum of less-to-more policy integration, going from intersectoral action (IA) to healthy public policy (HPP) to health in all policies (HiAP). Our case study included 34 municipal projects of the Dutch Gezonde Slagkracht Programme (2009-15), which supports the development and implementation of IPHP on overweight, alcohol and drug abuse, and smoking. Our content analysis of project application forms and interviews with all project leaders used a framework approach involving the policy strategies and the following policy variables: initiator, actors, policy goals, determinants and policy instruments. Most projects showed a combination of policy strategies. However, manifestations of IPHP in overweight projects predominantly involved IA. More policy integration was apparent in alcohol/drugs projects (HPP) and in all-theme projects (HiAP). More policy integration was related to broad goal definitions, which allowed for the involvement of actors representing several policy sectors. This enabled the implementation of a mix of policy instruments. Determinants of health were not explicitly used as a starting point of the policy process. If a policy problem justifies policy integration beyond IA, it might be helpful to start from the determinants of health (epidemiological reality), systematically transform them into policy (policy reality) and set broad policy goals, since this gives actors from other sectors the opportunity to participate.


The evolution of the Anopheles 16 genomes project.
We report the imminent completion of a set of reference genome assemblies for 16 species of Anopheles mosquitoes. In addition to providing a generally useful resource for comparative genomic analyses, these genome sequences will greatly facilitate exploration of the capacity exhibited by some Anopheline mosquito species to serve as vectors for malaria parasites. A community analysis project will commence soon to perform a thorough comparative genomic investigation of these newly sequenced genomes. Completion of this project via the use of short next-generation sequence reads required innovation in both the bioinformatic and laboratory realms, and the resulting knowledge gained could prove useful for genome sequencing projects targeting other unconventional genomes.


Iron status and the acute post-exercise hepcidin response in athletes.
This study explored the relationship between serum ferritin and hepcidin in athletes. Baseline serum ferritin levels of 54 athletes from the control trial of five investigations conducted in our laboratory were considered; athletes were grouped according to values <30 μg/L (SF<30), 30-50 μg/L (SF30-50), 50-100 μg/L (SF50-100), or >100 μg/L (SF>100). Data pooling resulted in each athlete completing one of five running sessions: (1) 8 × 3 min at 85% vVO2peak; (2) 5 × 4 min at 90% vVO2peak; (3) 90 min continuous at 75% vVO2peak; (4) 40 min continuous at 75% vVO2peak; (5) 40 min continuous at 65% vVO2peak. Athletes from each running session were represented amongst all four groups; hence, the mean exercise duration and intensity were not different (p>0.05). Venous blood samples were collected pre-, post- and 3 h post-exercise, and were analysed for serum ferritin, iron, interleukin-6 (IL-6) and hepcidin-25. Baseline and post-exercise serum ferritin levels were different between groups (p<0.05). There were no group differences for pre- or post-exercise serum iron or IL-6 (p>0.05). Post-exercise IL-6 was significantly elevated compared to baseline within each group (p<0.05). Pre- and 3 h post-exercise hepcidin-25 was sequentially greater as the groups baseline serum ferritin levels increased (p<0.05). However, post-exercise hepcidin levels were only significantly elevated in three groups (SF30-50, SF50-100, and SF>100; p<0.05). An athlete's iron stores may dictate the baseline hepcidin levels and the magnitude of post-exercise hepcidin response. Low iron stores suppressed post-exercise hepcidin, seemingly overriding any inflammatory-driven increases.