Generation of transepithelial potentials by isolated perfused reptilian distal tubules.
Transepithelial potentials were measured in the most distal segments of garter snake (Thamnophis spp.) distal tubles perfused in vitro. The segments generated high lumen-negative potentials when sodium was in the lumen. The size of the potentials was a saturable function of luminal sodium concentrations between 0 and 32 mM. The potentials were stable with time only when sodium concentrations in the lumen were less than 30 mM. Perfusion with high sodium concentrations resulted in transient potentials. Stable potentials changed markedly when the lumen sodium concentration or the bath potassium concentration was altered suddenly, but they were independent of lumen potassium concentrations and bath sodium concentrations. Amiloride stimulated or inhibited the potentials, ouabain partially depressed them, and ethacrynic acid and cyanide inhibited them slowly and often irreversibly. We conclude that distal transepithelial potentials reflect sodium transport from lumen to bath across a tight asymmetric epithelium which differs from other sodium-transporting epithelia in that stable transepithelial potentials are maintained only with luminal sodium concentrations less than 30 mM.


Religious differences in female genital cutting: a case study from Burkina Faso.
The relationship between religious obligations and female genital cutting is explored using data from Burkina Faso, a religiously and ethnically diverse country where approximately three-quarters of adult women are circumcised. Data from the 2003 Burkina Faso Demographic and Health Survey are used to estimate multilevel models of religious variation in the intergenerational transmission of female genital cutting. Differences between Christians, Muslims, and adherents of traditional religions are reported, along with an assessment of the extent to which individual and community characteristics account for religious differences. Religious variation in the intergenerational transmission of female genital cutting is largely explained by specific religious beliefs and by contextual rather than individual characteristics. Although Muslim women are more likely to have their daughters circumcised, the findings suggest the importance of a collective rather than individual Muslim identity for the continuation of the practice.


Detection of condensed-phase explosives via laser-induced vaporization, photodissociation, and resonant excitation.
We investigate the remote detection of explosives via a technique that vaporizes and photodissociates the condensed-phase material and detects the resulting vibrationally excited NO fragments via laser-induced fluorescence. The technique utilizes a single 7 ns pulse of a tunable laser near 236.2 nm to perform these multiple processes. The resulting blue-shifted fluorescence (226 nm) is detected using a photomultiplier and narrowband filter that strongly block the scatter of the pump laser off the solid media while passing the shorter wavelength photons. Various nitro-bearing compounds, including 2,6-dinitrotoluene (DNT), 2,4,6-trinitrotoluene (TNT), pentaerythritol tetranitrate (PETN), and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) were detected with a signal-to-noise of 25 dB. The effects of laser fluence, wavelength, and sample morphology were examined.


Plant-derived immunomodulators: an insight on their preclinical evaluation and clinical trials.
The phagocyte-microbe interactions in the immune system is a defense mechanism but when excessively or inappropriately deployed can harm host tissues and participate in the development of different non-immune and immune chronic inflammatory diseases such as autoimmune problems, allergies, some rheumatoid disorders, cancers and others. Immunodrugs include organic synthetics, biological agents such as cytokines and antibodies acting on single targets or pathways have been used to treat immune-related diseases but with limited success. Most of immunostimulants and immunosuppressants in clinical use are the cytotoxic drugs which possess serious side effects. There is a growing interest to use herbal medicines as multi-component agents to modulate the complex immune system in the prevention of infections rather than treating the immune-related diseases. Many therapeutic effects of plant extracts have been suggested to be due to their wide array of immunomodulatory effects and influence on the immune system of the human body. Phytochemicals such as flavonoids, polysaccharides, lactones, alkaloids, diterpenoids and glycosides, present in several plants, have been reported to be responsible for the plants immunomodulating properties. Thus the search for natural products of plant origin as new leads for development of potent and safe immunosuppressant and immunostimulant agents is gaining much major research interest. The present review will give an overview of widely investigated plant-derived compounds (curcumin, resveratrol, epigallocatechol-3-gallate, quercetin, colchicine, capsaicin, andrographolide, and genistein) which have exhibited potent effects on cellular and humoral immune functions in pre-clinical investigations and will highlight their clinical potential.


The effect of chlorhexidine dentifrice or gel versus chlorhexidine mouthwash on plaque, gingivitis, bleeding and tooth discoloration: a systematic review.
To systematically review and evaluate the available scientific evidence on the effectiveness of chlorhexidine dentifrice or gel (CHX DF/gel) compared to chlorhexidine mouthwash (CHX MW) on plaque, bleeding, gingival inflammation and tooth discoloration scores. PubMed-MEDLINE, Cochrane-CENTRAL and EMBASE databases were searched to identify appropriate studies. Independent screening of the 2256 unique titles and abstracts resulted in five publications that met the eligibility criteria. Considerable heterogeneity was found between the studies. Three of the five studies showed a positive effect on plaque scores in favour of the CHX MW. With respect to gingival index and bleeding scores, no significant differences were found. Chlorhexidine mouthwash, however, showed a significantly more tooth discoloration than the CHX DF/gel. A meta-analysis of the effect on 'de novo' plaque formation of CHX DF/gel versus CHX MW resulted in a difference in means of 0.27 [95% CI: 0.14; 0.39] (P < 0.0001). Chlorhexidine gel can be successfully formulated and will inhibit plaque growth to some degree, but not to the same extent, as a CHX MW. When CHX DF/gel is used in a non-brushing model, it is significantly less effective in plaque inhibition compared to CHX MW. Based on one study when CHX gel was applied with a finger after brushing, it is significantly more effective on plaque scores and the gingival index. The only brushing study also with a long follow-up showed that there is no significant difference between CHX DF and CHX MW. However, as a corollary, significantly more tooth discoloration was observed with the CHX MW. Altogether, the data show that when daily oral hygiene cannot be performed, CHX MW is the first product of choice.


Understanding and measuring quality of care: dealing with complexity.
Existing definitions and measurement approaches of quality of health care often fail to address the complexities involved in understanding quality of care. It is perceptions of quality, rather than clinical indicators of quality, that drive service utilization and are essential to increasing demand. Here we reflect on the nature of quality, how perceptions of quality influence health systems and what such perceptions indicate about measurement of quality within health systems. We discuss six specific challenges related to the conceptualization and measurement of the quality of care: perceived quality as a driver of service utilization; quality as a concept shaped over time through experience; responsiveness as a key attribute of quality; the role of management and other so-called upstream factors; quality as a social construct co-produced by families, individuals, networks and providers; and the implications of our observations for measurement. Within the communities and societies where care is provided, quality of care cannot be understood outside social norms, relationships, trust and values. We need to improve not only technical quality but also acceptability, responsiveness and levels of patient-provider trust. Measurement approaches need to be reconsidered. An improved understanding of all the attributes of quality in health systems and their interrelationships could support the expansion of access to essential health interventions.


Prophylactic Bilateral Nipple-sparing Mastectomy and a Staged Breast Reconstruction Technique: Preliminary Results.
More high-risk women with breast cancer are identified using genetic testing at a younger age. These young women often opt for prophylactic surgery. Most patients are reluctant for extra donor-site scars besides infections and necrosis. In order to reduce these risks, a two-stage breast reconstruction technique is used for high-risk women with large or ptotic breasts. We presume that this procedure will reduce the risk of skin envelope and nipple-areola complex (NAC) necrosis to less than 1%. In the first stage, an inferior pedicle reduction is performed to obtain large volume reduction with maximal safety for the NAC. The ptosis, skin excess, and malpositioning of the NAC are corrected safely at this stage. In the second stage, the skin-sparing mastectomy is performed with or without nipple sparing. During this procedure, the areola is never removed. A bilateral breast reconstruction is then performed with an immediate subpectoral prothesis or delayed with the use of a subpectoral tissue expander. In this way, we aim to meet the patient's wish to undergo bilateral risk reducing mastectomy in breasts that need ptosis correction without donor-site scarring. This article describes the procedure and reports the preliminary data.


Catecholaminergic depletion within the prelimbic medial prefrontal cortex enhances latent inhibition.
Latent inhibition (LI) refers to the reduction in conditioning to a stimulus that has received repeated non-reinforced pre-exposure. Investigations into the neural substrates of LI have focused on the nucleus accumbens (NAc) and its inputs from the hippocampal formation and adjacent cortical areas. Previous work has suggested that lesions to the medial prefrontal cortex (mPFC), another major source of input to the NAc, do not disrupt LI. However, a failure to observe disrupted LI does not preclude the possibility that a particular brain region is involved in the expression of LI. Moreover, the mPFC is a heterogeneous structure and there has been no investigation of a possible role of different regions within the mPFC in regulating LI under conditions that prevent LI in controls. Here, we tested whether 6-hydroxydopamine (6-OHDA)-induced lesions of dopamine (DA) terminals within the prelimbic (PL) and infralimbic (IL) mPFC would lead to the emergence of LI under conditions that do produce LI in controls (weak pre-exposure). LI was measured in a thirst motivated conditioned emotional response procedure with 10 pre-exposures to a noise conditioned stimulus (CS) and two conditioning trials. Sham-operated and IL-lesioned animals did not show LI and conditioned to the pre-exposed CS at comparable levels to the non-pre-exposed controls. 6-OHDA lesions to the PL, however, produced potentiation of LI. These results provide the first demonstration that the PL mPFC is a component of the neural circuitry underpinning LI.


Emotional development across adulthood: differential age-related emotional reactivity and emotion regulation in a negative mood induction procedure.
The present study examines the hypothesis that older adults might differentially react to a negative versus neutral mood induction procedure than younger adults. The rationale for this expectation was derived from Socioemotional Selectivity Theory (SST), which postulates differential salience of emotional information and ability to regulate emotions across adulthood. The present data support a view of differential age-related effects of negative mood inductions with greater and more heterogeneous emotional reactivity among older adults, who showed a substantially greater decrease in self-rated pleasantness, calmness, and wakefulness than younger adults. Moreover, relative to the younger adults, emotion regulation in terms of mood repair was more effective among the older adults. The age-related mood effects are discussed in terms of SST and have practical implications for the study of emotion and cognition across adulthood.


Systematic Review of Criminal and Legal Involvement After Substance Use and Mental Health Treatment Among Veterans: Building Toward Needed Research.
Evidence indicates that substance use and mental health treatment is often associated with reduced criminal activity. The present systematic review examined this association among military veterans, and aimed to provide a comprehensive summary of needed research to further contribute to reduced criminal activity among veterans. This systematic review was derived from a scoping review that mapped existing research on justice-involved veterans' health. For the current systematic review, a subset of 20 publications was selected that addressed the question of whether criminal activity declines among veterans treated for substance use and mental health disorders. Generally, veterans improved on criminal outcomes from pre- to post-treatment for opioid use, other substance use, or mental health conditions, and more sustained treatment was associated with better outcomes. This occurred despite high rates of criminal involvement among veterans prior to entering treatment. Needed are substance use and mental health treatment studies that include women justice-involved veterans, follow criminally-active veterans for longer periods of time, and use validated and reliable measures of criminal activity with fully transparent statistical procedures. Future randomized trials should evaluate new treatments against evidence-based treatments (versus no-treatment control conditions). Subsequent studies should examine how to link veterans to effective treatments, facilitate sustained treatment engagement, and ensure the availability of effective treatments, and examine mechanisms (mediators and moderators) that explain the association of treatment with reduced criminal activity among veterans. Best practices are needed for reducing criminal activity among the minority of justice-involved veterans who do not have diagnosed substance use and/or mental health disorders.


Intrinsic survival mechanisms for retinal ganglion cells.
Retinal ganglion cells (RGCs) undergo programmed cell death (apoptosis) after axonal injury. This cell death is mediated by several mechanisms, including deprivation of neurotrophic factors, alterations in gene expression, and production of reactive oxygen species. However, death of RGCs is delayed after axonal injury, and a significant number survive even after several days. This suggests that RGC death is not an immediate result of axonal injury, and that other pro-survival factors may play a role. While we and other researchers have focused on the mechanisms of cell death after axonal injury, it may be that determining the regulation of cell survival mechanisms may lead to innovative methods for neuroprotection. The final common pathway of glaucomatous optic neuropathy is RGC death, probably via damage to their axons occurring at or near the lamina cribrosa. Axonal injury leads directly (1) or indirectly (2) to the death of retinal ganglion cells. We and others have demonstrated that axotomy is associated with RGC apoptosis (3-7) as well as specific changes in expression of certain genes at the mRNA and protein level (8, 9). Reactive oxygen species may also be part of the pathway for RGC death (10, 11). We therefore hypothesize that axotomy leads to molecular events that are potentially destructive to RGCs, but also induces changes that are potentially protective against cellular injury. If this is the case, then RGC death from axonal injury would result not only from initiation of apoptosis, but also from failure of intrinsic neuroprotective mechanisms. It should therefore be theoretically possible to modulate these two classes of responses, and thus improve RGC cell survival after axotomy.


The wheat resistance gene Lr34 results in the constitutive induction of multiple defense pathways in transgenic barley.
The wheat gene Lr34 encodes an ABCG-type transporter which provides durable resistance against multiple pathogens. Lr34 is functional as a transgene in barley, but its mode of action has remained largely unknown both in wheat and barley. Here we studied gene expression in uninfected barley lines transgenic for Lr34. Genes from multiple defense pathways contributing to basal and inducible disease resistance were constitutively active in seedlings and mature leaves. In addition, the hormones jasmonic acid and salicylic acid were induced to high levels, and increased levels of lignin as well as hordatines were observed. These results demonstrate a strong, constitutive re-programming of metabolism by Lr34. The resistant Lr34 allele (Lr34res) encodes a protein that differs by two amino acid polymorphisms from the susceptible Lr34sus allele. The deletion of a single phenylalanine residue in Lr34sus was sufficient to induce the characteristic Lr34-based responses. Combination of Lr34res and Lr34sus in the same plant resulted in a reduction of Lr34res expression by 8- to 20-fold when the low-expressing Lr34res line BG8 was used as a parent. Crosses with the high-expressing Lr34res line BG9 resulted in an increase of Lr34sus expression by 13- to 16-fold in progenies that inherited both alleles. These results indicate an interaction of the two Lr34 alleles on the transcriptional level. Reduction of Lr34res expression in BG8 crosses reduced the negative pleiotropic effects of Lr34res on barley growth and vigor without compromising disease resistance, suggesting that transgenic combination of Lr34res and Lr34sus can result in agronomically useful resistance.


Protein deacetylation by SIRT1: an emerging key post-translational modification in metabolic regulation.
The biological function of most proteins relies on reversible post-translational modifications, among which phosphorylation is most prominently studied and well recognized. Recently, a growing amount of evidence indicates that acetylation-deacetylation reactions, when applied to crucial mediators, can also robustly affect the function of target proteins and thereby have wide-ranging physiological impacts. Sirtuin 1 (SIRT1), which functions as a nicotinamide adenine dinucleotide (NAD(+))-dependent protein deacetylase, deacetylates a wide variety of metabolic molecules in response to the cellular energy and redox status and as such causes significant changes in metabolic homeostasis. This review surveys the evidence for the emerging role of SIRT1-mediated deacetylation in the control of metabolic homeostasis.


Correlates of Burnout in Small Independent Primary Care Practices in an Urban Setting.
Little is known about the prevalence and correlates of burnout among providers who work in small independent primary care practices (<5 providers). We conducted a cross-sectional analysis by using data collected from 235 providers practicing in 174 small independent primary care practices in New York City. The rate of provider-reported burnout was 13.5%. Using bivariate logistic regression, we found higher adaptive reserve scores were associated with lower odds of burnout (odds ratio, 0.12; 95% CI, 0.02-0.85; P = .034). The burnout rate was relatively low among our sample of providers compared with previous surveys that focused primarily on larger practices. The independence and autonomy providers have in these small practices may provide some protection against symptoms of burnout. In addition, the relationship between adaptive reserve and lower rates of burnout point toward potential interventions for reducing burnout that include strengthening primary care practices' learning and development capacity.


Explaining impossible phenomena: object permanence beliefs and memory failures in adults.
In three experiments, adult subjects' explanations of the observed nonpermanence of a physical object and their recollections of the order of the events during the experiment were obtained and analysed. The data showed that in order to conserve their strong beliefs in object permanence subjects systematically distorted the real temporal succession of events preceding the phenomenon. The frequency of the distortions depended on the salience of the nonpermanence phenomenon ("disappearance" versus "appearance" of the physical object) and on the time interval between the events whose temporal order was reversed, but not on subjects' nationality (English versus German), gender, type of reproduction (immediate versus delayed), role in the experiment (subject versus observer), and degree of prompting in questioning.


Stopped-flow spectrophotometric analysis of intermediates in the peroxo-dependent inactivation of cytochrome P450 by aldehydes.
The reaction of hydrogen peroxide and certain aromatic aldehydes with cytochrome P450BM3-F87G results in the covalent modification of the heme cofactor of this monooxygenase. Analysis of the resulting heme by electronic absorption spectrophotometry indicates that the reaction in the BM3 isoform is analogous to that in P450(2B4), which apparently occurs via a peroxyhemiacetal intermediate [Kuo et al., Biochemistry, 38 (1999) 10511]. It was observed that replacement of the Phe-87 in the P450BM3 by the smaller glycyl residue was essential for the modification to proceed, as the wild-type enzyme showed no spectral changes under identical conditions. The kinetics of this reaction were examined by stopped-flow spectrophotometry with 3-phenylpropionaldehyde and 3-phenylbutyraldehyde as reactants. In each case, the process of heme modification was biphasic, with initial bleaching of the Soret absorbance, followed by an increase in absorbance centered at 430 nm, consistent with meso-heme adduct formation. The intermediate formed during phase I also showed an increased absorbance between 700 and 900 nm, relative to the native heme and the final product. Phase I showed a linear dependence on peroxide concentration, whereas saturation kinetics were observed for phase II. All of these observations are consistent with a mechanism involving radical attack at the gamma-meso position of the heme cofactor, resulting in the intermediate formation of an isoporphyrin, the deprotonation of which produces the gamma-meso-alkyl heme derivative.


Individual Differences in Children's (Language) Learning Skills Moderate Effects of Robot-Assisted Second Language Learning.
The current study investigated how individual differences among children affect the added value of social robots for teaching second language (L2) vocabulary to young children. Specifically, we investigated the moderating role of three individual child characteristics deemed relevant for language learning: first language (L1) vocabulary knowledge, phonological memory, and selective attention. We expected children low in these abilities to particularly benefit from being assisted by a robot in a vocabulary training. An L2 English vocabulary training intervention consisting of seven sessions was administered to 193 monolingual Dutch five-year-old children over a three- to four-week period. Children were randomly assigned to one of three experimental conditions: 1) a tablet only, 2) a tablet and a robot that used deictic (pointing) gestures (the no-iconic-gestures condition), or 3) a tablet and a robot that used both deictic and iconic gestures (i.e., gestures depicting the target word; the iconic-gestures condition). There also was a control condition in which children did not receive a vocabulary training, but played dancing games with the robot. L2 word knowledge was measured directly after the training and two to four weeks later. In these post-tests, children in the experimental conditions outperformed children in the control condition on word knowledge, but there were no differences between the three experimental conditions. Several moderation effects were found. The robot's presence particularly benefited children with larger L1 vocabularies or poorer phonological memory, while children with smaller L1 vocabularies or better phonological memory performed better in the tablet-only condition. Children with larger L1 vocabularies and better phonological memory performed better in the no-iconic-gestures condition than in the iconic-gestures condition, while children with better selective attention performed better in the iconic-gestures condition than the no-iconic-gestures condition. Together, the results showed that the effects of the robot and its gestures differ across children, which should be taken into account when designing and evaluating robot-assisted L2 teaching interventions.


Corticotropin-Releasing Hormone Receptor Type 1 (CRHR1) Clustering with MAGUKs Is Mediated via Its C-Terminal PDZ Binding Motif.
The corticotropin-releasing hormone receptor type 1 (CRHR1) plays an important role in orchestrating neuroendocrine, behavioral, and autonomic responses to stress. To identify molecules capable of directly modulating CRHR1 signaling, we performed a yeast-two-hybrid screen using the C-terminal intracellular tail of the receptor as bait. We identified several members of the membrane-associated guanylate kinase (MAGUK) family: postsynaptic density protein 95 (PSD95), synapse-associated protein 97 (SAP97), SAP102 and membrane associated guanylate kinase, WW and PDZ domain containing 2 (MAGI2). CRHR1 is co-expressed with the identified MAGUKs and with the additionally investigated PSD93 in neurons of the adult mouse brain and in primary hippocampal neurons, supporting the probability of a physiological interaction in vivo. The C-terminal PDZ (PSD-95, discs large, zona occludens 1) binding motif of CRHR1 is essential for its physical interaction with MAGUKs, as revealed by the CRHR1-STAVA mutant, which harbors a functionally impaired PDZ binding motif. The imitation of a phosphorylation at Thr413 within the PDZ binding motif also disrupted the interaction with MAGUKs. In contrast, distinct PDZ domains within the identified MAGUKs are involved in the interactions. Expression of CRHR1 in primary neurons demonstrated its localization throughout the neuronal plasma membrane, including the excitatory post synapse, where the receptor co-localized with PSD95 and SAP97. The co-expression of CRHR1 and respective interacting MAGUKs in HEK293 cells resulted in a clustered subcellular co-localization which required an intact PDZ binding motif. In conclusion, our study characterized the PDZ binding motif-mediated interaction of CRHR1 with multiple MAGUKs, which directly affects receptor function.


Risk factors for and outcomes of noncatastrophic suspensory apparatus injury in Thoroughbred racehorses.
To evaluate effects of toe grabs, exercise intensity, and distance traveled as risk factors for subclinical to mild suspensory apparatus injury (SMSAI) in Thoroughbred racehorses and to compare incidence of severe musculoskeletal injury (MSI) in horses with and without SMSAI. Nested case-control study. 219 Thoroughbred racehorses racing or in race training. Racehorses were examined weekly for 90 days to determine incidence of suspensory ligament injury and monitor horseshoe characteristics. Every horse's exercise speeds and distances were recorded daily. Conditional logistic regression was used to compare exposure variables between incident case (n = 25) and selected control (125) horses. Survival analysis was used to compare time to MSI for horses with (n = 41) and without (76) SMSAI. The best-fitting logistic model for the data included age (< 5 vs > or = 5 years old), toe grab height the week of injury (none vs very low, low, regular, or Quarter Horse height), and weekly distance the week preceding injury (miles). Although the 95% confidence intervals for all odds ratios included 1, the odds for SMSAI appeared to increase with the presence of a toe grab, higher weekly distance, and age > or = 5 years. Horses that had SMSAI were significantly more likely to have a severe MSI or severe suspensory apparatus injury than were horses that did not. Results suggest that pre-existing SMSAI is associated with development of severe MSI and severe suspensory apparatus injury. Modifying training intensity and toe grab height for horses with SMSAI may decrease the incidence of severe MSI.


Data Augmentation in High Dimensional Low Sample Size Setting Using a Geometry-Based Variational Autoencoder.
In this paper, we propose a new method to perform data augmentation in a reliable way in the High Dimensional Low Sample Size (HDLSS) setting using a geometry-based variational autoencoder (VAE). Our approach combines the proposal of 1) a new VAE model, the latent space of which is modeled as a Riemannian manifold and which combines both Riemannian metric learning and normalizing flows and 2) a new generation scheme which produces more meaningful samples especially in the context of small data sets. The method is tested through a wide experimental study where its robustness to data sets, classifiers and training samples size is stressed. It is also validated on a medical imaging classification task on the challenging ADNI database where a small number of 3D brain magnetic resonance images (MRIs) are considered and augmented using the proposed VAE framework. In each case, the proposed method allows for a significant and reliable gain in the classification metrics. For instance, balanced accuracy jumps from 66.3% to 74.3% for a state-of-the-art convolutional neural network classifier trained with 50 MRIs of cognitively normal (CN) and 50 Alzheimer disease (AD) patients and from 77.7% to 86.3% when trained with 243 CN and 210 AD while improving greatly sensitivity and specificity metrics.


Genome-based prediction of common diseases: advances and prospects.
Common diseases such as type 2 diabetes and coronary heart disease result from a complex interplay of genetic and environmental factors. Recent developments in genomics research have boosted progress in the discovery of susceptibility genes and fueled expectations about opportunities of genetic profiling for personalizing medicine. Personalized medicine requires a test that fairly accurately predicts disease risk, particularly when interventions are invasive, expensive or have major side effects. Recent studies on the prediction of common diseases based on multiple genetic variants alone or in addition to traditional disease risk factors showed limited predictive value so far, but all have investigated only a limited number of susceptibility variants. New gene discoveries from genome-wide association studies will certainly further improve the prediction of common diseases, but the question is whether this improvement is sufficient to enable personalized medicine. In this paper, we argue that new gene discoveries may not evidently improve the prediction of common diseases to a degree that it will change the management of individuals at increased risk. Substantial improvements may only be expected if we manage to understand the complete causal mechanisms of common diseases to a similar extent as we understand those of monogenic disorders. Genomics research will contribute to this understanding, but it is likely that the complexity of complex diseases may ultimately limit the opportunities for accurate prediction of disease in asymptomatic individuals as unraveling their complete causal pathways may be impossible.


Polychlorinated biphenyls disturb differentiation of normal human neural progenitor cells: clue for involvement of thyroid hormone receptors.
Polychlorinated biphenyls (PCBs) are ubiquitous environmental chemicals that accumulate in adipose tissues over the food chain. Epidemiologic studies have indicated that PCBs influence brain development. Children who are exposed to PCBs during development suffer from neuropsychologic deficits such as a lower full-scale IQ (intelligence quotient), reduced visual recognition memory, and attention and motor deficits. The mechanisms leading to these effects are not fully understood. It has been speculated that PCBs may affect brain development by interfering with thyroid hormone (TH) signaling. Because most of the data are from animal studies, we established a model using primary normal human neural progenitor (NHNP) cells to determine if PCBs interfere with TH-dependent neural differentiation. NHNP cells differentiate into neurons, astrocytes, and oligodendrocytes in culture, and they express a variety of drug metabolism enzymes and nuclear receptors. Like triiodothyronine (T3), treatment with the mono-ortho-substituted PCB-118 (2,3',4,4 ,5-pentachlorobiphenyl; 0.01-1 microM) leads to a dose-dependent increase of oligodendrocyte formation. This effect was congener specific, because the coplanar PCB-126 (3,3',4,4 ,5-pentachlorobiphenyl) had no effect. Similar to the T3 response, the PCB-mediated effect on oligodendrocyte formation was blocked by retinoic acid and the thyroid hormone receptor antagonist NH-3. These results suggest that PCB-118 mimics T3 action via the TH pathway.


Acquired ventricular septal defect: a rare sequel of blunt chest trauma in a 7-year-old boy.
Ventricular septal defect (VSD) is the most common congenital cardiac lesion encountered worldwide. Only very rarely is it acquired, and causation through blunt injury in a child is extremely rare. A previously healthy 7-year-old boy suffered blunt chest trauma while at play. He presented 11 days later with features of acute congestive cardiac failure. Two-dimensional echocardiographic examination revealed a mid-muscular VSD. The connection between the defect and the trauma was not initially appreciated. Facilities for required urgent open-heart surgery were not available. Cardiac failure was refractory to anti-failure therapy. His clinical condition steadily worsened, and he succumbed after 20 days on admission. We conclude that a diagnosis of traumatic VSD, though rare, should be considered in any previously well child presenting in acute congestive cardiac failure following blunt trauma to the chest. Any such patient should undergo careful echocardiographic evaluation. There is an urgent need for facilities for open-heart surgery to be more readily available and accessible in Nigeria.


Functional Analysis of a Novel FOXL2 Indel Mutation in Chinese Families with Blepharophimosis-Ptosis-Epicanthus Inversus Syndrome Type I.
Background: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is an autosomal dominant disease with a low incidence rate. Indel mutations in the forkhead box L2 (FOXL2) gene cause two types of BPES that are distinguished by the presence (type I) or absence (type II) of premature ovarian failure (POF). The purpose of this study was to identify a possible deletion in FOXL2 in Chinese families with BPES and to clarify its relationship with POF. Methods: An autosomal dominant Chinese BPES family with four generations was enrolled in this study. Peripheral venous blood was collected from all affected patients, and genomic DNA was extracted from leukocytes. The whole coding sequence and nearby 5' untranslated region (UTR) and 3'UTR of the FOXL2 gene were amplified using polymerase chain reaction (PCR) with three sets of overlapping primers, followed by sequencing analyses. The sequencing results were analysed using SeqMan software. Based on the patients' clinical manifestations and analysis of the identified indel mutation, we found that the mutation disturbed interactions between FOXL2 and the StAR gene. Furthermore, through subcellular localisation and functional studies, we observed significant mislocalisation of the mutant protein; the mutant protein was found in the cytoplasm, while the wild-type protein was found in the nucleus. Loss of function was confirmed by transcriptional activity assays, quantitative real-time PCR, and electrophoretic mobility shift assays. Results: All affected patients presented with clinical features of BPES type I, including small palpebral fissures, ptosis, telecanthus, and epicanthus inversus with POF. A novel FOXL2 heterozygous indel mutation, c.19_95del, a 77-bp deletion that disrupts FOXL2 protein structure, was identified in all affected members of the family. In addition, this indel mutation significantly increased StAR mRNA expression by disrupting the ability of the FOXL2 protein to bind to the StAR promoter and act as a repressor of this gene. Conclusions: A novel FOXL2 indel mutation was identified in Chinese families with BPES. Our results expand the spectrum of known FOXL2 mutations and provide additional insight into the structure-function relationships of the FOXL2 protein. Furthermore, this novel mutation resulted in the dysfunction of FOXL2 as a transcription factor, blocking its ability to bind to the promoter region of the StAR gene, resulting in POF in the affected patient.


Safety and diagnostic value of brain biopsy in HIV patients: a case series and meta-analysis of 1209 patients.
Early brain biopsy may be indicated in HIV patients with focal brain lesion. This study aimed to evaluate and compare the safety and diagnostic value of brain biopsy in HIV patients in the pre-highly active antiretroviral therapy (HAART) versus post-HAART era via meta-analysis. Appropriate studies were identified per search criteria. The local database was retrospectively reviewed to select a similar patient cohort. Patient demographics, brain biopsy technique, histopathology and patient outcomes were extracted from each study. Study-specific outcomes were combined per random-effects model. Outcomes were compared between the pre-HAART and post-HAART era. Correlations between outcomes and baseline characteristics were assessed via meta-regression analysis. The proportions of histopathological diagnosis were tabulated and compared between the pre- and post-HAART era. Survival analysis was performed for patients in the post-HAART era. A total of 26 studies (including the local database) with 1209 patients were included in this meta-analysis. The most common indications for brain biopsy were diagnosis unlikely to be toxoplasmosis (n=8, 42.1%), focal brain lesion (n=5, 26.3%) or both (n=3, 15.8%). The weighted proportions for diagnostic success were 92% (95% CI 90.0% to 93.8%), change in management 57.7% (45.9% to 69.1%) and clinical improvement 36.6% (26.3% to 47.5%). Morbidity and mortality were 5.7% (3.6% to 8.3%) and 0.9% (0.3% to 1.9%), respectively. Diagnostic success rate was significantly higher in the post-HAART than the pre-HAART era (97.5% vs 91.9%, p=0.047). The odds ratio (OR) for diagnostic success in patients with contrast-enhanced lesions was 2.54 ((1.25 to 5.15), p<0.01). The median survival for HIV patients who underwent biopsy in the post-HAART era was 225 days (90-2446). Brain biopsy in HIV patients is safe with high diagnostic yield. Early brain biopsy should be considered in patients without classic presentation of toxoplasmosis encephalitis.


Dissociating Attention Effects from Categorical Perception with ERP Functional Microstates.
When faces appear in our visual environment we naturally attend to them, possibly to the detriment of other visual information. Evidence from behavioural studies suggests that faces capture attention because they are more salient than other types of visual stimuli, reflecting a category-dependent modulation of attention. By contrast, neuroimaging data has led to a domain-specific account of face perception that rules out the direct contribution of attention, suggesting a dedicated neural network for face perception. Here we sought to dissociate effects of attention from categorical perception using Event Related Potentials. Participants viewed physically matched face and butterfly images, with each category acting as a target stimulus during different blocks in an oddball paradigm. Using a data-driven approach based on functional microstates, we show that the locus of endogenous attention effects with ERPs occurs in the N1 time range. Earlier categorical effects were also found around the level of the P1, reflecting either an exogenous increase in attention towards face stimuli, or a putative face-selective measure. Both category and attention effects were dissociable from one another hinting at the role that faces may play in early capturing of attention before top-down control of attention is observed. Our data support the conclusion that certain object categories, in this experiment, faces, may capture attention before top-down voluntary control of attention is initiated.


Enhanced self-assembly for the solubilization of cholesterol in molecular solvent/ionic liquid mixtures.
The development of new solvents combining greatly enhanced solubility for sparingly soluble compounds and good kinetic properties is challenging. In this study, we constructed a family of new molecular solvent/ionic liquid (IL) mixtures with amphiphilic, anionic functional long-chain carboxylate ionic liquids (LCC-ILs) as a key component for the solubilization of sparingly soluble compounds, using cholesterol as a model solute. Polarized optical microscopy (POM), wide angle X-ray diffraction (WAXD), Fourier-transform infrared (FTIR) spectra and 1H NMR showed that ordered mesoscopic structures, such as liquid crystals (LCs), were formed when cholesterol was dissolved in the mixtures, presenting a self-assembly induced dissolution mechanism driven by H-bond interaction and van der Waals forces in the mixtures. A synergistic effect between the molecular solvents and LCC-ILs was revealed, which contributed to enhanced solute-solvent self-assembly in dissolution over pure LCC-ILs and thus elevated solubility. Additionally, the effect of IL concentration, solvent type and anionic alkyl-chain length on self-assembly and solubility was investigated. These mixtures showed unparalleled solubilities for cholesterol, while maintaining a low viscosity. The quantitative solubilities (g g-1) of cholesterol were as high as 0.70, 0.84 and 0.82, respectively, at 25 °C in ethyl acetate/[P4444][C15H31COO] (50 wt%), n-heptane/[P4444][C15H31COO] (40 wt%) and ethyl acetate/[P4444][C17H35COO] (50 wt%) mixtures, which were the highest solubilities of cholesterol ever reported, six- to 980-fold higher than traditional molecular solvents and even one- to seven-fold higher compared to pure LCC-ILs. These results demonstrated the considerable potential of molecular solvent/LCC-ILs mixtures as promising solvents for solubilization and advanced separation processes.


Colloidal electroconvection in a thin horizontal cell. II. Bulk electroconvection of water during parallel-plate electrolysis.
We recently have reported [J. Chem. Phys. 122, 164701 (2005)] a family of electroconvective patterns that arise when charge-stabilized colloidal dispersions are driven by constant (dc) vertical electric fields. Competition between gravity and electrokinetic forces acting on the individual spheres in this system leads to the formation of highly organized convective instabilities involving hundreds of spheres. Here, we report a distinct class of electroconvective patterns that emerge in confined aqueous dispersions at higher biases. These qualitatively resemble the honeycomb and labyrinthine patterns formed during thermally driven Rayleigh-Benard convection, but arise from a distinct mechanism. Unlike the localized colloidal electroconvective patterns observed at lower biases, moreover, these system-spanning patterns form even without dispersed colloidal particles. Rather, they appear to result from an underlying electroconvective instability during electrolysis in the parallel plate geometry. This contrasts with recent theoretical results suggesting that simple electrolytes are linearly stable against electroconvection.


Patency of the posterior communicating artery following treatment with the Pipeline Embolization Device.
OBJECTIVE The Pipeline Embolization Device (PED) has become an effective treatment strategy for some cerebral aneurysms. Concerns regarding the patency of branch arteries have been raised. The objective of this study was to assess the patency of the posterior communicating artery (PCoA) following treatment of PCoA aneurysms using the PED. METHODS All patients with PCoA aneurysms treated with the PED who had angiographic follow-up were retrospectively identified. The patency of the PCoA at follow-up was evaluated by 2 authors who were not involved in the intervention. Univariate and multivariate analyses were performed to identify factors associated with the following: 1) PCoA patency versus no or diminished flow, and 2) PCoA patency and diminished flow versus PCoA occlusion. RESULTS Thirty patients with an angiographic follow-up of 6 months were included. Aneurysm obliteration was achieved in 25 patients (83.3%). The PCoA was patent in 7 patients (23.3%), had diminished flow in 7 patients (23.3%), and was occluded in 16 patients (53.3%). In the univariate analysis of outcome, there was a trend for aneurysms with incomplete occlusion, aneurysms not previously treated, those with presence of a fetal PCoA, and those with an artery coming from the aneurysm to have higher odds of the PCoA remaining patent. In univariate and multivariate analyses of factors associated with outcome, fetal PCoA and presence of an artery coming from the aneurysm were associated with the PCoA remaining open with or without diminished flow. No patients had symptoms related to PCoA occlusion. CONCLUSIONS Occlusion and diminished flow through the PCoA is common following PED treatment of PCoA aneurysms. However, it is clinically insignificant in most cases.


Peroxisome proliferator-activated receptor-gamma regulates the expression of alveolar macrophage macrophage colony-stimulating factor.
Macrophage CSF (M-CSF) regulates monocyte differentiation, activation, and foam cell formation. We have observed that it is elevated in human pulmonary alveolar proteinosis (PAP) and in the GM-CSF knockout mouse, a murine model for PAP. A potential regulator of M-CSF, peroxisome proliferator-activated receptor-gamma (PPARgamma), is severely deficient in both human PAP and the GM-CSF knockout mouse. To investigate the role of PPARgamma in alveolar macrophage homeostasis, we generated myeloid-specific PPARgamma knockout mice using the Lys-Cre method to knock out the floxed PPARgamma gene. Similar to the GM-CSF-deficient mouse, absence of alveolar macrophage PPARgamma resulted in development of lung pathology resembling PAP in 16-wk-old mice, along with excess M-CSF gene expression and secretion. In ex vivo wild-type alveolar macrophages, we observed that M-CSF itself is capable of inducing foam cell formation similar to that seen in PAP. Overexpression of PPARgamma prevented LPS-stimulated M-CSF production in RAW 264.7 cells, an effect that was abrogated by a specific PPARgamma antagonist, GW9662. Use of proteasome inhibitor, MG-132 or a PPARgamma agonist, pioglitazone, prevented LPS-mediated M-CSF induction. Using chromatin immunoprecipitation, we found that PPARgamma is capable of regulating M-CSF through transrepression of NF-kappaB binding at the promoter. Gel-shift assay experiments confirmed that pioglitazone is capable of blocking NF-kappaB binding. Taken together, these data suggest that M-CSF is an important mediator of alveolar macrophage homeostasis, and that transcriptional control of M-CSF production is regulated by NF-kappaB and PPARgamma.


Behaviors of susceptible-infected epidemics on scale-free networks with identical infectivity.
In this paper, we propose a susceptible-infected model with identical infectivity, in which, at every time step, each node can only contact a constant number of neighbors. We implemented this model on scale-free networks, and found that the infected population grows in an exponential form with the time scale proportional to the spreading rate. Furthermore, by numerical simulation, we demonstrated that the targeted immunization of the present model is much less efficient than that of the standard susceptible-infected model. Finally, we investigate a fast spreading strategy when only local information is available. Different from the extensively studied path-finding strategy, the strategy preferring small-degree nodes is more efficient than that preferring large-degree nodes. Our results indicate the existence of an essential relationship between network traffic and network epidemic on scale-free networks.


The Frequency and Subjective Impact of Painful and Provocative Events on the Acquired Capability for Suicide.
Leading theoretical explanations for suicide state that for suicide death to occur, a person must have sufficient capability to enact an attempt. Exposure to painful and provocative events is hypothesized to play an important role in acquiring the capability for suicide over a lifetime. Unfortunately, assessment tools for painful and provocative events have focused solely on the frequency of events, neglecting the potential contributions of perceived impact. Further, past measurements have used predetermined items for painful and provocative events thereby neglecting other relevant events. The current study uses visual analog scales (VAS) to assess both the frequency and impact of painful and provocative events and how these contribute to the capability for suicide. Data were collected from 787 adults via Amazon's online platform. Findings indicated that the frequency VAS and impact VAS both had a moderate correlation with the original Painful and Provocative Events Scale. Greater scores on the frequency VAS were associated with increased capability, whereas lower scores on the subjective impact VAS were associated with increased capability scores. Both VAS independently predicted capability above and beyond the PPES. Temporal or causal associations are unable to be drawn due to cross-sectional data. In addition, the sample was largely homogenous (White = 72%, female = 63.5%), limiting generalizability. These initial findings demonstrate individuals who perceive painful and provocative events as being less impactful may have increased capability, and that VAS may be appropriate to approximate the impact and frequency of painful and provocative events.


Facile Synthesis of Molecularly Imprinted Graphene Quantum Dots for the Determination of Dopamine with Affinity-Adjustable.
A facilely prepared fluorescence sensor was developed for dopamine (DA) determination based on polyindole/graphene quantum dots molecularly imprinted polymers (PIn/GQDs@MIPs). The proposed sensor exhibits a high sensitivity with a linear range of 5 × 10(-10) to 1.2 × 10(-6) M and the limit of detection as low as 1 × 10(-10) M in the determination of DA, which is probably due to the tailor-made imprinted cavities for binding DA thought hydrogen bonds between amine groups of DA and oxygen-containing groups of the novel composite. Furthermore, the prepared sensor can rebind DA in dual-type: a low affinity type (noncovalent interaction is off) and a high affinity type (noncovalent interaction is on), and the rebinding interaction can be adjusted by tuning the pH, which shows a unique potential for adjusting the binding interaction while keeping the specificity, allowing for wider applications.


The effect of therapeutic touch on behavioral symptoms of persons with dementia.
Approximately 80% of nursing home residents who suffer from Alzheimer's disease and related dementia develop behavioral symptoms of dementia. Given the deleterious side effects of pharmacologic therapy in this population there is an urgent need for clinical trials of nonpharmacologic interventions. To examine the effect of therapeutic touch on the frequency and intensity of behavioral symptoms of dementia. A randomized, double-blind, three-group experimental study: experimental (therapeutic touch), placebo (placebo therapeutic touch), and control (usual care). Fifty-seven residents, aged 67 to 93 years, exhibiting behavioral symptoms of dementia, were randomized to one of the three groups within each of three Special Care Units within three Long-Term Care facilities in a western Canadian province. Behavioral observation was completed every 20 minutes from 8:00AM to 6:00PM for three days pre-intervention and for three days post-intervention by trained observers who were blind to group assignment. The intervention consisted of therapeutic touch given twice daily for 5-7 minutes for three days between 10:00AM and 11:30PM and between 3:00PM and 4:30PM (N = 57). The main outcome variable was overall behavioral symptoms of dementia, consisting of six categories of behaviors: manual manipulation (restlessness), escape restraints, searching and wandering, tapping and banging, pacing and walking, and vocalization. Analysis of variance (ANOVA) (F = 3.331, P = .033) and the Kruskal-Wallis test (chi2 = 6.661, P = .036) indicated a significant difference in overall behavioral symptoms of dementia, manual manipulation and vocalization when the experimental group was compared to the placebo and control groups. The experimental (significant) was more effective in decreasing behavioral symptoms of dementia than usual care, while the placebo group indicated a decreasing trend in behavioral symptoms of dementia compared to usual care. Therapeutic touch offers a nonpharmacological, clinically relevant modality that could be used to decrease behavioral symptoms of dementia, specifically manual manipulation (restlessness) and vocalization, two prevalent behaviors.


Studies of the effectiveness of interferon alpha treatment for chronic hepatitis C in children.
The significance of hepatitis C infection in Poland, particularly in a pathology of the developmental age still increased. The aim of the study was the analysis of interferon alpha therapy efficacy in children with chronic C hepatitis. 30 children (aged from 3 years to 15 years, 16 females, 14 males) were included in the study. In each patient HCV infection was confirmed by the serological, molecular (with identification of HCV genotype) and histopathological methods. The duration of observation of HCV-infected children after the diagnosis was made followed for at least 6 months. Transaminase level in each case was 50% higher than normal. The schema of interferon alpha treatment was: 3 MU 3 times a week subcutaneously for 25 weeks. Time of observation started at the beginning of the therapy and finished 1 year after the end of the treatment. The analysis of the HCV genotypes showed the predominance of the genotype 1 (66.7%): subtype 1a was found in 20% patients, subtype 1b--in 43.5% children. Genotype 4 (subtype 4c4d or 4b) was confirmed in 30% patients, genotype 3 (subtype 3a) in 3.3% patients. In the histopathological picture of the liver predominated minimal or moderate inflammation activity (grading: 1--in 50%, 2--in 46.6%, 3--in 3.4%) and little fibrosis (staging: 0--in 80%, 1--in 13.3%, 2--in 6.7%). In many children mild side effects of interferon alpha therapy were observed: pseudoinfluenzal symptoms (in 46.7%), lack of appetite (in 16.7%), abdominal pain (in 10%), thrombocytopenia (in 6.7%), granulocytopenia, hair loss, irritability, itching of the skin (in 3.4%). At the end of therapy in 36.7% patients serum HCV-RNA was undetectable. The percentage of children without serum HCV-RNA decreased 6 months after the end of therapy to 20% patients and a year after the end of therapy to only 13.6% children. In children with HCV-RNA elimination was observed early reduction of ALT level. For the definition of the predictive factors of good prognosis patients were divided into 2 groups: group I (without HCV-RNA elimination at the end of the treatment) and group II (patients HCV-RNA negative a year after the end of therapy). Both group of children were similar in respect of age, disease duration and interferon alpha dosis/m2. At the beginning of the treatment mean ALT level was statistically higher in group II than in group I. IL-2 level was significant higher in group II than at the beginning, after 2 and 4 months of the therapy. There were no significant differences in IL-1 beta, IL-4 and IL-6 level between patients in group I and II. The differences in ALT activity during IFN-therapy between 2 groups of patients were statistically significant; since second month of therapy higher ALT level was observed in a group of patients without HCV-RNA elimination. In the histopathological picture of the liver a year of the end of therapy in 20% children reduction of inflammatory activity and progression of fibrosis in both group of patients was observed. Because of a little efficacy, high costs, psychological load of young patients and possible following consequences of the interferon alpha therapy it is necessary to manage the further researches to find a new method of treatment of chronic C hepatitis. High ALT activity and elevated IL-2 level before treatment seems to be predictive factors of the good response to interferon alpha therapy.


The Carter Center's assistance to river blindness control programs: establishing treatment objectives and goals for monitoring ivermectin delivery systems on two continents.
Periodic mass treatment with ivermectin in endemic communities prevents eye and dermal disease due to onchocerciasis. As part of an international global partnership to control onchocerciasis, The Carter Center's Global 2000 River Blindness Program (GRBP) assists the ministries of health in ten countries to distribute ivermectin (Mectizan, donated by Merck & Co.). The GRBP priorities are to maximize ivermectin treatment coverage and related health education and training efforts, and to monitor progress through regular reporting of ivermectin treatments measured against annual treatment objectives and ultimate treatment goals (e.g., full coverage, which is defined as reaching all persons residing in at risk villages who are eligible for treatment). Since the GRBP began in 1996, more than 21.2 million ivermectin treatment encounters have been reported by assisted programs. In 1999, more than 6.6 million eligible persons at risk for onchocerciasis received treatment, which represented 96% of the 1999 annual treatment objective of 6.9 million, and 78% of the ultimate treatment goal in assisted areas.


E-cadherin as an epithelial barrier protein in exhaled breath condensate.
Airway epithelium plays an important role as a physical barrier and a modulator of allergic response. Junctions between cells provide epithelial integrity and barrier function. The aim of this study was to investigate the influence of atopy on airway epithelial integrity in asthma and to measure E-cadherin levels in exhaled breath condensate as an indicator epithelial damage. A total of 74 patients with asthma (35 atopic and 39 non-atopic) and 39 healthy children were enrolled in this case-control study. Sociodemographic characteristics and asthma severity parameters in the last three-month period were recorded and pulmonary function tests were performed. Blood samples were obtained to measure serum immunoglobulin E (IgE) levels and peripheral blood eosinophil count, and exhaled breath condensate (EBC) was obtained to measure E-cadherin.EBC E-cadherin levels were significantly lower in the asthmatics when compared to non-atopic controls (0.109 (0.076) versus 0.191 (0.184) ng mL(-1) respectively, p = 0.01). Atopic and non-atopic asthmatic groups had lower EBC E-cadherin levels compared to the control group. (0.112 (0.060) ng ml(-1), 0.106 (0.089) ng ml(-1) and 0.191 (0.184) ng ml(-1), p = 0.02 and p < 0.01 respectively). However, EBC E-cadherin levels were not different between atopic and non-atopic asthmatics. The results of our study support the role of E-cadherin in the pathogenesis of asthma. However, the absence of difference in E-cadherin levels between atopic and non-atopic asthmatics suggests that allergic sensitization is not the primary factor for development of epithelial barrier dysfunction in asthma.


Preparation and characterization of inorganic radioactive holmium-166 microspheres for internal radionuclide therapy.
Microspheres with high specific activities of radionuclides are very interesting for internal radiotherapy treatments. This work focuses on the formulation and characterization of inorganic microspheres with a high content of holmium and therefore a high specific radioactivity of holmium-166. Two novel formulations of inorganic microspheres were obtained by dispersing solid holmium acetylacetonate microspheres (Ho2(AcAc)3-ms) in NaH2PO4 or NaOH solutions followed by 2 h incubation at room temperature. By exchange of acetylacetonate with phosphate or hydroxyl ions, holmium phosphate microspheres (HoPO4-ms) and holmium hydroxide microspheres (Ho(OH)3-ms) were formed respectively. The inorganic microspheres had a significantly smaller diameter (28.5 ± 4.4 μm (HoPO4-ms) and 25.1 ± 3.5 μm (Ho(OH)3-ms)) than those of Ho2(AcAc)3-ms (32.6 ± 5.2 μm). The weight percentage of holmium-165 in the microspheres increased significantly from 47% (Ho2(AcAc)3-ms) to 55% (HoPO4-ms) and 73% (Ho(OH)3-ms). After preparation of both HoPO4-ms and Ho(OH)3-ms, the stable holmium-165 isotope was partly converted by neutron activation into radioactive holmium-166 to yield radioactive microspheres. High specific activities were achieved ranging from 21.7 to 59.9 MBq/mg (166HoPO4-ms) and from 28.8 to 79.9 MBq/mg (166Ho(OH)3-ms) depending on the neutron activation time. The structure of both microspheres was preserved up to neutron activations of 6 h in a thermal neutron flux of 4.72 × 1016 n m-2 s-1. After activation, both microspheres revealed excellent stability in administration fluids (saline and phosphate buffer) having less than 0.05% of holmium released after 72 h incubation. Finally, the hemocompatibility of these inorganic microspheres was evaluated and it was shown that the microspheres did cause neither hemolysis nor depletion or inhibition of the coagulation factors of the intrinsic blood coagulation pathway meaning that the microspheres have a good hemocompatibility. Overall, this work shows that radioactive inorganic microspheres with high specific activities of holmium-166 can be prepared which potentially can be used for internal radionuclide therapy.


Adolescent vs. adult onset of a first episode psychosis: Impact on remission of positive and negative symptoms.
Adolescent-onset psychosis has traditionally been characterized as a more severe form of psychosis with a poorer prognosis. However, it is still unclear if patients with an adolescent-onset have worse symptom remission outcomes. Symptom remission is the principal clinical outcome known to predict quality of life and social functioning in the long term. The goal of this study is to clarify the influence of age of onset of psychosis on symptom remission in a sample of first-episode psychosis patients. A total of 246 first-episode psychosis patients were recruited from a specialized early intervention program serving a defined epidemiological catchment area. Age of onset of psychosis (adolescence vs. adulthood) was used as the main predictor, and duration of untreated psychosis (DUP), baseline symptoms, baseline functioning, substance abuse diagnosis, medication adherence and gender were used as covariates in hierarchical regression models predicting the following positive and negative symptom remission outcomes: maximum continuous months in remission and early remission (i.e., occurring in the first three months of follow-up). After controlling for other variables, onset of psychosis in adulthood and shorter DUP predicted early remission of positive symptoms. This effect was stronger in patients with a diagnosis of a schizophrenia-spectrum disorder. Remission of negative symptoms did not depend on age of onset, and was only predicted by baseline negative symptoms. Patients with onset of psychosis during adulthood are more likely to achieve early positive symptom remission than those with adolescent onset. This effect might be stronger in patients with a diagnosis of a schizophrenia-spectrum disorder.


A New Natural Product Analog of Blasticidin S Reveals Cellular Uptake Facilitated by the NorA Multidrug Transporter.
The permeation of antibiotics through bacterial membranes to their target site is a crucial determinant of drug activity but in many cases remains poorly understood. During screening efforts to discover new broad-spectrum antibiotic compounds from marine sponge samples, we identified a new analog of the peptidyl nucleoside antibiotic blasticidin S that exhibited up to 16-fold-improved potency against a range of laboratory and clinical bacterial strains which we named P10. Whole-genome sequencing of laboratory-evolved strains of Staphylococcus aureus resistant to blasticidin S and P10, combined with genome-wide assessment of the fitness of barcoded Escherichia coli knockout strains in the presence of the antibiotics, revealed that restriction of cellular access was a key feature in the development of resistance to this class of drug. In particular, the gene encoding the well-characterized multidrug efflux pump NorA was found to be mutated in 69% of all S. aureus isolates resistant to blasticidin S or P10. Unexpectedly, resistance was associated with inactivation of norA, suggesting that the NorA transporter facilitates cellular entry of peptidyl nucleosides in addition to its known role in the efflux of diverse compounds, including fluoroquinolone antibiotics.


Earthing the human body influences physiologic processes.
This study was designed to answer the question: Does the contact of the human organism with the Earth via a copper conductor affect physiologic processes? Subjects and experiments: Five (5) experiments are presented: experiment 1-effect of earthing on calcium-phosphate homeostasis and serum concentrations of iron (N = 84 participants); experiment 2-effect of earthing on serum concentrations of electrolytes (N = 28); experiment 3-effect of earthing on thyroid function (N = 12); experiment 4-effect of earthing on glucose concentration (N = 12); experiment 5-effect of earthing on immune response to vaccine (N = 32). Subjects were divided into two groups. One (1) group of people was earthed, while the second group remained without contact with the Earth. Blood and urine samples were examined. Earthing of an electrically insulated human organism during night rest causes lowering of serum concentrations of iron, ionized calcium, inorganic phosphorus, and reduction of renal excretion of calcium and phosphorus. Earthing during night rest decreases free tri-iodothyronine and increases free thyroxine and thyroid-stimulating hormone. The continuous earthing of the human body decreases blood glucose in patients with diabetes. Earthing decreases sodium, potassium, magnesium, iron, total protein, and albumin concentrations while the levels of transferrin, ferritin, and globulins α1, α2, β, and γ increase. These results are statistically significant. Earthing the human body influences human physiologic processes. This influence is observed during night relaxation and during physical activity. Effect of the earthing on calcium-phosphate homeostasis is the opposite of that which occurs in states of weightlessness. It also increases the activity of catabolic processes. It may be the primary factor regulating endocrine and nervous systems.


[Angiotensin converting enzyme inhibitors, angiotensin receptor blockers and combined therapy in microalbuminuric patients with one or more cardiovascular risk factors. Protocol of the Long-term Impact of RAS Inhibition on Cardiorenal Outcomes randomized trial (LIRICO).].
Angiotensin converting enzyme inhibitors (ACE-i) and angiotensin II receptor blockers (ARB) are considered to be equally effective for patients with diabetic kidney disease, while only ACE-i have been shown to determine a significant reduction in the risk of all-cause mortality, predominantly cardiovascular, in these patients. Studies on the cardio-renal efficacy of combined therapy with ACE-i and ARB are not available or not conclusive, in a population with cardiovascular risk with micro- or macroalbuminuria. In this paper, we present the protocol of a randomized controlled clinical trial that will address the question. The LIRICO (Long-term Impact of RAS Inhibition on Cardiorenal Outcomes) study will evaluate the comparative efficacy for cardiovascular and renal outcomes of combined therapy with ACE-i and ARB versus monotherapy with ACE-i or ARB in micro/macroalbuminuric individuals at cardio-renal risk. The study will enrol 2100 patients allocated to monotherapy with ACE-i, ARB or combined treatment with ACE-i + ARB. The LIRICO study is a randomized comparative trial, with PROBE (Prospective Randomized Open Blinded End-Point) design. The study has been approved and funded by the Agenzia Italiana del Farmaco (AIFA) within the 2005 funding plan for independent research on drugs. Availability of funding for this study provides, for the first time in our Country, an opportunity to organize a collaborative national network of nephrology, internal medicine and diabetology outpatient clinics to develop a large multicentre trial collaboration. The results of this trial will establish the optimal therapy for micro/macroalbuminuric individuals with cardiovascular and renal risk.


Weekend and nighttime effect on the prognosis of peptic ulcer bleeding.
To evaluate whether weekend or nighttime admission affects prognosis of peptic ulcer bleeding despite early endoscopy. Retrospective data collection from four referral centers, all of which had a formal out-of-hours emergency endoscopy service, even at weekends. A total of 388 patients with bleeding peptic ulcers who were admitted via the emergency room between January 2007 and December 2009 were enrolled. Analyzed parameters included time from patients' arrival until endoscopy, mortality, rebleeding, need for surgery and length of hospital stay. The weekday and weekend admission groups comprised 326 and 62 patients, respectively. There were no significant differences in baseline characteristics between the two groups, except for younger age in the weekend group. Most patients (97%) had undergone early endoscopy, which resulted in a low mortality rate regardless of point of presentation (1.8% overall vs 1.6% on the weekend). The only outcome that was worse in the weekend group was a higher rate of rebleeding (12% vs 21%, P = 0.030). However, multivariate analysis revealed nighttime admission and a high Rockall score (≥ 6) as significant independent risk factors for rebleeding, rather than weekend admission. Early endoscopy for peptic ulcer bleeding can prevent the weekend effect, and nighttime admission was identified as a novel risk factor for rebleeding, namely the nighttime effect.


Impact of work schedules on sleep duration of critical care nurses.
Sleep deprivation leads to reduced vigilance and potentially impairs work performance. Nurses may work long shifts that may contribute to sleep deprivation. To assess how nurses' sleep patterns are affected by work schedules and other factors. Between October 2009 and June 2010, a total of 20 critical care nurses completed daily sleep and activity logs and a demographic survey and wore an actigraph to objectively measure sleep time for 14 days. In a multivariate model with controls for repeated measures, mean sleep time between consecutive work shifts was short: 6.79 hours between 2 day shifts and 5.68 hours between 2 night shifts (P = .01). Sleep time was much greater between days when no shifts were worked (8.53 hours), consistent with catch-up sleep during these times. Every minute of 1-way commuting time was associated with a reduction of sleep time by 0.84 minutes. Critical care nurses obtain reduced amounts of sleep between consecutive work shifts, particularly between consecutive night shifts. Whether this degree of sleep deprivation adversely affects patients' safety needs further study.


Matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of bacteria isolated from pharmaceutical clean rooms.
During the manufacturing of sterile drugs, it is of the utmost importance to meet the minimum requirements for asepsis recommended by the legislations on good manufacturing practices-based efficient environmental monitoring. The availability of relatively simple to use matrix-assisted laser desorption ionization-time of flight mass spectromtomy (MALDI-TOF MS) devices in the last years has changed the laboratory workflows for the microbial identification, mainly in the clinical area. Thus, the objective of this work was to evaluate the suitability of the MALDI-TOF MS technique for the identification of bacteria isolated from the environment of clean rooms used in some stages of the production of a viral vaccine. Eighteen known bacterial species commonly isolated from clean rooms studied were identified by MALDI-TOF technique and by a biochemical technique (BBL Crystal® System). Performance of MALDI-TOF MS was better than biochemical technique for correct species identifications (88.89% and 38.89%, respectively) and produced less unreliable identification (5.55% and 22.22%). MALDI-TOF MS can be implemented for routine identification of bacteria in a pharmaceutical quality control laboratory, but as a database-dependent system, maybe some isolated not identified by this technique must be additionally studied and, if appropriate, added to an in-house database.


De novo genome assembly of the economically important weed horseweed using integrated data from multiple sequencing platforms.
Horseweed (Conyza canadensis), a member of the Compositae (Asteraceae) family, was the first broadleaf weed to evolve resistance to glyphosate. Horseweed, one of the most problematic weeds in the world, is a true diploid (2n = 2x = 18), with the smallest genome of any known agricultural weed (335 Mb). Thus, it is an appropriate candidate to help us understand the genetic and genomic bases of weediness. We undertook a draft de novo genome assembly of horseweed by combining data from multiple sequencing platforms (454 GS-FLX, Illumina HiSeq 2000, and PacBio RS) using various libraries with different insertion sizes (approximately 350 bp, 600 bp, 3 kb, and 10 kb) of a Tennessee-accessed, glyphosate-resistant horseweed biotype. From 116.3 Gb (approximately 350× coverage) of data, the genome was assembled into 13,966 scaffolds with 50% of the assembly = 33,561 bp. The assembly covered 92.3% of the genome, including the complete chloroplast genome (approximately 153 kb) and a nearly complete mitochondrial genome (approximately 450 kb in 120 scaffolds). The nuclear genome is composed of 44,592 protein-coding genes. Genome resequencing of seven additional horseweed biotypes was performed. These sequence data were assembled and used to analyze genome variation. Simple sequence repeat and single-nucleotide polymorphisms were surveyed. Genomic patterns were detected that associated with glyphosate-resistant or -susceptible biotypes. The draft genome will be useful to better understand weediness and the evolution of herbicide resistance and to devise new management strategies. The genome will also be useful as another reference genome in the Compositae. To our knowledge, this article represents the first published draft genome of an agricultural weed.


A high-throughput chemically induced inflammation assay in zebrafish.
Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening. Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction. This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay. See Commentary article: http://www.biomedcentral.com/1741-7007/8/148.


Functions of noncoding sequences in mammalian genomes.
Most of the mammalian genome consists of nucleotide sequences not coding for proteins. Exons of genes make up only 3% of the human genome, while the significance of most other sequences remains unknown. Recent genome studies with high-throughput methods demonstrate that the so-called noncoding part of the genome may perform important functions. This hypothesis is supported by three groups of experimental data: 1) approximately 10% of the sequences, most of which are located in noncoding parts of the genome, is evolutionarily conserved and thus can be of functional importance; 2) up to 99% of the mammalian genome is being transcribed forming short and long noncoding RNAs in addition to common mRNA; and 3) mutations in noncoding parts of the genome can be accompanied by progression of pathological states of the organism. In the light of these data, in the review we consider the functional role of numerous known sequences of noncoding parts of the genome including introns, DNA methylation regions, enhancers and locus control regions, insulators, S/MAR sequences, pseudogenes, and genes of noncoding RNAs, as well as transposons and simple repeats of centromeric and telomeric regions of chromosomes. The assumption is made that the intergenic noncoding sequences without definite/clear functions can be involved in spatial organization of genetic loci in interphase nuclei.


Viral etiology, seasonality and severity of hospitalized patients with severe acute respiratory infections in the Eastern Mediterranean Region, 2007-2014.
Little is known about the role of viral respiratory pathogens in the etiology, seasonality or severity of severe acute respiratory infections (SARI) in the Eastern Mediterranean Region. Sentinel surveillance for SARI was conducted from December 2007 through February 2014 at 20 hospitals in Egypt, Jordan, Oman, Qatar and Yemen. Nasopharyngeal and oropharyngeal swabs were collected from hospitalized patients meeting SARI case definitions and were analyzed for infection with influenza, respiratory syncytial virus (RSV), adenovirus (AdV), human metapneumovirus (hMPV) and human parainfluenza virus types 1-3 (hPIV1-3). We analyzed surveillance data to calculate positivity rates for viral respiratory pathogens, describe the seasonality of those pathogens and determine which pathogens were responsible for more severe outcomes requiring ventilation and/or intensive care and/or resulting in death. At least one viral respiratory pathogen was detected in 8,753/28,508 (30.7%) samples tested for at least one pathogen and 3,497/9,315 (37.5%) of samples tested for all pathogens-influenza in 3,345/28,438 (11.8%), RSV in 3,942/24,503 (16.1%), AdV in 923/9,402 (9.8%), hMPV in 617/9,384 (6.6%), hPIV1 in 159/9,402 (1.7%), hPIV2 in 85/9,402 (0.9%) and hPIV3 in 365/9,402 (3.9%). Multiple pathogens were identified in 501/9,316 (5.4%) participants tested for all pathogens. Monthly variation, indicating seasonal differences in levels of infection, was observed for all pathogens. Participants with hMPV infections and participants less than five years of age were significantly less likely than participants not infected with hMPV and those older than five years of age, respectively, to experience a severe outcome, while participants with a pre-existing chronic disease were at increased risk of a severe outcome, compared to those with no reported pre-existing chronic disease. Viral respiratory pathogens are common among SARI patients in the Eastern Mediterranean Region. Ongoing surveillance is important to monitor changes in the etiology, seasonality and severity of pathogens of interest.


Model structure and control of bone remodeling: a theoretical study.
It is generally accepted that RANKL is highly expressed in osteoblast precursor cells while OPG is highly expressed in mature osteoblasts, but to date no functional utility to the BMU has been proposed for this particular ligand-decoy-receptor expression profile. As discovered in the mid 90s, the RANK-RANKL-OPG signaling cascade is a major signaling pathway regulating bone remodeling. In this paper we study theoretically the functional implications of particular RANKL/OPG expression profiles on bone volume. For this purpose we formulate an extended bone-cell dynamics model describing functional behaviour of basic multicellular units (BMUs) responsible for bone resorption and formation. This model incorporates the RANK-RANKL-OPG signaling together with the regulating action of TGF-beta on bone cells. The bone-cell population model employed here builds on the work of Lemaire et al. (2004) [1], but incorporates the following significant modifications: (i) addition of a rate equation describing changes in bone volume with time as the key 'output function' tracking functional behaviour of BMUs, (ii) a rate equation describing release of TGF-beta from the bone matrix, (iii) expression of OPG and RANKL on both osteoblastic cell lines, and (iv) modified activator/repressor functions. Using bone volume as a functional selection criterion, we find that there is a preferred arrangement for ligand expression on particular cell types, and further, that this arrangement coincides with biological observations. We then investigate the model parameter space combinatorially, searching for preferred 'groupings' of changes in differentiation rates of various cell types. Again, a criterion of bone volume change is employed to identify possible ways of optimally controlling BMU responses. While some combinations of changes in differentiation rates are clearly unrealistic, other combinations of changes in differentiation rates are potentially functionally significant. Most importantly, the combination of parameter changes representing the signaling pathway for TGF-beta gives a unique result that appears to have a clear biological rationale. The methodological approach for the investigation of model structure described here offers a theoretical explanation as to why TGF-beta has its particular suite of biological effects on bone-cell differentiation rates.


Recent advances in primary cutaneous T-cell lymphoma.
Cutaneous T-cell lymphoma (CTCL) is a heterogeneous group of skin-homing T-cell neoplasms, which represent approximately 75% of all primary cutaneous lymphomas. Currently available drug therapies, when effective, simply control disease and the only option for curing CTCL is stem cell transplant. In the last year, there has been an incredible effort made to improve the understanding and treatment of CTCL. Recent findings indicate that epigenetic aberrations are integral to active disease. Furthermore, multiple tumor-derived immunological factors have also been shown to inhibit viability, proliferation, and cytokine production of nonmalignant T cells. Several novel targeted therapies show great potential, most promising being antibody drug conjugates targeting surface markers such as CD30 in some CTCL subtypes. Additional attractive targets involve the global modulation of epigenetic markers such as demethylation agents or HDAC inhibitors, either as single agents or in combination therapies. This is a concise review of recent advances in the field of CTCL with special focus on research articles over the preceding year.


Seizure prediction - ready for a new era.
Epilepsy is a common disorder characterized by recurrent seizures. An overwhelming majority of people with epilepsy regard the unpredictability of seizures as a major issue. More than 30 years of international effort have been devoted to the prediction of seizures, aiming to remove the burden of unpredictability and to couple novel, time-specific treatment to seizure prediction technology. A highly influential review published in 2007 concluded that insufficient evidence indicated that seizures could be predicted. Since then, several advances have been made, including successful prospective seizure prediction using intracranial EEG in a small number of people in a trial of a real-time seizure prediction device. In this Review, we examine advances in the field, including EEG databases, seizure prediction competitions, the prospective trial mentioned and advances in our understanding of the mechanisms of seizures. We argue that these advances, together with statistical evaluations, set the stage for a resurgence in efforts towards the development of seizure prediction methodologies. We propose new avenues of investigation involving a synergy between mechanisms, models, data, devices and algorithms and refine the existing guidelines for the development of seizure prediction technology to instigate development of a solution that removes the burden of the unpredictability of seizures.


Reactivity of platinum alkyne complexes towards N-fluorobenzenesulfonimide: formation of platinum compounds bearing a β-fluorovinyl ligand.
The platinum(0) alkyne complexes [Pt(L)(η(2)-PhC[triple bond, length as m-dash]CPh)] 1-4 were synthesized by reactions of [Pt(cod)2] with diphenylacetylene and a phosphine ligand precursor (1: L = dcpe, 2: L = xantphos, 3: L = κ(2)-(P,N)-iPr2PC3H6NMe2, 4: L = κ(2)-(P,N)-iPr2PC2H4NMe2). Treatment of 1 or 4 with NFSI gave the complexes [Pt(F){N(SO2Ph)2}(dcpe)] (5) and [Pt(PhC[double bond, length as m-dash]CFPh){N(SO2Ph)2}{κ(2)-(P,N)-iPr2PC2H4NMe2}] (8), whereas the reactivity of 2 and 3 towards NFSI led to product mixtures. The compounds [Pt(F){N(SO2Ph)2}(xantphos)] (6a) as well as [Pt(PhC[double bond, length as m-dash]CFPh){N(SO2Ph)2}{κ(2)-(P,N)-iPr2PC2H4NMe2}] (7a) and [Pt(PhC[double bond, length as m-dash]CFPh)(F){κ(2)-(P,N)-iPr2PC2H4NMe2}] (7b) were clearly identified. Ligand exchange reactions at 8 resulted in the formation of the β-fluorovinyl platinum(ii) complexes [Pt(PhC[double bond, length as m-dash]CFPh){OC(O)CF3}{κ(2)-(P,N)-iPr2PC2H4NMe2}] (9), [Pt(PhC[double bond, length as m-dash]CFPh)(FHF){κ(2)-(P,N)-iPr2PC2H4NMe2}] (10) and [Pt(PhC[double bond, length as m-dash]CFPh)(F){κ(2)-(P,N)-iPr2PC2H4NMe2}] (11). Treatment of 8 with dihydrogen yielded the fluorinated olefin (Z)-(1-fluoroethene-1,2-diyl)dibenzene and [Pt{N(SO2Ph)2}(H){κ(2)-(P,N)-iPr2PC2H4NMe2}] (12).


An ultra-broadband multilayered graphene absorber.
An ultra-broadband multilayered graphene absorber operating at terahertz (THz) frequencies is proposed. The absorber design makes use of three mechanisms: (i) The graphene layers are asymmetrically patterned to support higher order surface plasmon modes that destructively interfere with the dipolar mode and generate electromagnetically induced absorption. (ii) The patterned graphene layers biased at different gate voltages backed-up with dielectric substrates are stacked on top of each other. The resulting absorber is polarization dependent but has an ultra-broadband of operation. (iii) Graphene's damping factor is increased by lowering its electron mobility to 1000 cm²/Vs. Indeed, numerical experiments demonstrate that with only three layers, bandwidth of 90% absorption can be extended upto 7THz, which is drastically larger than only few THz of bandwidth that can be achieved with existing metallic/graphene absorbers.


ESHRE survey results and good practice recommendations on managing chromosomal mosaicism.
How should ART/preimplantation genetic testing (PGT) centres manage the detection of chromosomal mosaicism following PGT? Thirty good practice recommendations were formulated that can be used by ART/PGT centres as a basis for their own policy with regards to the management of 'mosaic' embryos. The use of comprehensive chromosome screening technologies has provided a variety of data on the incidence of chromosomal mosaicism at the preimplantation stage of development and evidence is accumulating that clarifies the clinical outcomes after transfer of embryos with putative mosaic results, with regards to implantation, miscarriage and live birth rates, and neonatal outcomes. This document was developed according to a predefined methodology for ESHRE good practice recommendations. Recommendations are supported by data from the literature, a large survey evaluating current practice and published guidance documents. The literature search was performed using PubMed and focused on studies published between 2010 and 2022. The survey was performed through a web-based questionnaire distributed to members of the ESHRE special interest groups (SIG) Reproductive Genetics and Embryology, and the ESHRE PGT Consortium members. It included questions on ART and PGT, reporting, embryo transfer policy and follow-up of transfers. The final dataset represents 239 centres. The working group (WG) included 16 members with expertise on the ART/PGT process and chromosomal mosaicism. The recommendations for clinical practice were formulated based on the expert opinion of the WG, while taking into consideration the published data and results of the survey. Eighty percent of centres that biopsy three or more cells report mosaicism, even though only 66.9% of all centres have validated their technology and only 61.8% of these have validated specifically for the calling of chromosomal mosaicism. The criteria for designating mosaicism, reporting and transfer policies vary significantly across the centres replying to the survey. The WG formulated recommendations on how to manage the detection of chromosomal mosaicism in clinical practice, considering validation, risk assessment, designating and reporting mosaicism, embryo transfer policies, prenatal testing and follow-up. Guidance is also provided on the essential elements that should constitute the consent forms and the genetic report, and that should be covered in genetic counselling. As there are several unknowns in chromosomal mosaicism, it is recommended that PGT centres monitor emerging data on the topic and adapt or refine their policy whenever new insights are available from evidence. Rather than providing instant standardized advice, the recommendations should help ART/PGT centres in developing their own policy towards the management of putative mosaic embryos in clinical practice. This document will help facilitate a more knowledge-based approach for dealing with chromosomal mosaicism in different centres. In addition to recommendations for clinical practice, recommendations for future research were formulated. Following up on these will direct research towards existing research gaps with direct translation to clinical practice. Emerging data will help in improving guidance, and a more evidence-based approach of managing chromosomal mosaicism. The WG received technical support from ESHRE. M.D.R. participated in the EQA special advisory group, outside the submitted work, and is the chair of the PGT WG of the Belgian society for human genetics. D.W. declared receiving salary from Juno Genetics, UK. A.C. is an employee of Igenomix, Italy and C.R. is an employee of Igenomix, Spain. C.S. received a research grant from FWO, Belgium, not related to the submitted work. I.S. declared being a Co-founder of IVFvision Ltd, UK. J.R.V. declared patents related to 'Methods for haplotyping single-cells' and 'Haplotyping and copy number typing using polymorphic variant allelic frequencies', and being a board member of Preimplantation Genetic Diagnosis International Society (PGDIS) and International Society for Prenatal Diagnosis (ISPD). K.S. reported being Chair-elect of ESHRE. The other authors had nothing to disclose. This Good Practice Recommendations (GPR) document represents the views of ESHRE, which are the result of consensus between the relevant ESHRE stakeholders and are based on the scientific evidence available at the time of preparation. ESHRE GPRs should be used for information and educational purposes. They should not be interpreted as setting a standard of care or be deemed inclusive of all proper methods of care, or be exclusive of other methods of care reasonably directed to obtaining the same results. They do not replace the need for application of clinical judgement to each individual presentation, or variations based on locality and facility type. Furthermore, ESHRE GPRs do not constitute or imply the endorsement, or favouring, of any of the included technologies by ESHRE.


Stromal cell inhibition of anti-CD20 antibody mediated killing of B-cell malignancies.
Introduction: The glycoengineered type II anti-CD20 monoclonal antibody obinutuzumab has been licensed for treatment in follicular non-Hodgkin lymphoma and B-CLL following clinical trials demonstrating superior outcomes to standard of care treatment. However, ultimately many patients still relapse, highlighting the need to understand the mechanisms behind treatment failure to improve patient care. Resistance to chemotherapy is often caused by the ability of malignant B-cells to migrate to the bone marrow and home into the stromal layer. Therefore, this study aimed to investigate whether stromal cells were also able to inhibit type II anti-CD20 antibody mechanisms of action, contributing to resistance to therapy. Methods: A stromal-tumor co-culture was established in vitro between Raji or Daudi B-cell tumor cells and M210B4 stromal cells in 24 well plates. Results: Contact with stromal cells was able to protect tumor cells from obinutuzumab mediated programmed cell death (PCD), antibody dependent cellular phagocytosis and antibody dependent cellular cytotoxicity. Furthermore, such protection required direct contact between stroma and tumor cells. Stromal cells appeared to interfere with obinutuzumab mediated B-cell homotypic adhesion through inhibiting and reversing actin remodelling, potentially as a result of stromal-tumor cell contact leading to downregulation of CD20 on the surface of tumor cells. Further evidence for the potential role of CD20 downregulation comes through the reduction in surface CD20 expression and inhibition of obinutuzumab mediated PCD when tumor cells are treated with Ibrutinib in the presence of stromal cells. The proteomic analysis of tumor cells after contact with stromal cells led to the identification of a number of altered pathways including those involved in cell adhesion and the actin cytoskeleton and remodeling. Discussion: This work demonstrates that contact between tumor cells and stromal cells leads to inhibition of Obinutuzumab effector functions and has important implications for future therapies to improve outcomes to anti-CD20 antibodies. A deeper understanding of how anti-CD20 antibodies interact with stromal cells could prove a useful tool to define better strategies to target the micro-environment and ultimately improve patient outcomes in B-cell malignancies.


Cyclooxygenase-2 is a novel transcriptional target of the nuclear EGFR-STAT3 and EGFRvIII-STAT3 signaling axes.
Emerging evidence indicates a novel mode of epidermal growth factor receptor (EGFR) signaling, notably, one involves EGFR nuclear translocalization and subsequent gene activation. To date, however, the significance of the nuclear EGFR pathway in glioblastoma (GBM) is unknown. Here, we report that EGFR and its constitutively activated variant EGFRvIII undergo nuclear translocalization in GBM cells, in which the former event requires EGF stimulation and the latter is constitutive. To gain insights into the effect of nuclear EGFR on gene expression in GBM, we created isogenic GBM cell lines, namely, U87MG-vector, U87MG-EGFR, and U87MG-EGFRdNLS that, respectively, express the control vector, EGFR, and nuclear entry-defective EGFR with a deletion of the nuclear localization signal (NLS). Microarray analysis shows that 19 genes, including cyclooxygenase-2 (COX-2), to be activated in U87MG-EGFR cells but not in U87MG-EGFRdNLS and U87MG-vector cells. Subsequent validation studies indicate that COX-2 gene is expressed at higher levels in cells with EGFR and EGFRvIII than those with EGFRdNLS and EGFRvIIIdNLS. Nuclear EGFR and its transcriptional cofactor signal transducer and activator of transcription 3 (STAT3) associate with the COX-2 promoter. Increased expression of EGFR/EGFRvIII and activated STAT3 leads to the synergistic activation of the COX-2 promoter. Promoter mutational analysis identified a proximal STAT3-binding site that is required for EGFR/EGFRvIII-STAT3-mediated COX-2 gene activation. In GBM tumors, an association exists between levels of COX-2, EGFR/EGFRvIII, and activated STAT3. Together, these findings indicate the existence of the nuclear EGFR/EGFRvIII signaling pathway in GBM and its functional interaction with STAT3 to activate COX-2 gene expression, thus linking EGFR-STAT3 and EGFRvIII-STAT3 signaling axes to proinflammatory COX-2 mediated pathway.