The transcription factor nerve growth factor-inducible protein a mediates epigenetic programming: altering epigenetic marks by immediate-early genes.
Maternal care alters epigenetic programming of glucocorticoid receptor (GR) gene expression in the hippocampus, and increased postnatal maternal licking/grooming (LG) behavior enhances nerve growth factor-inducible protein A (NGFI-A) transcription factor binding to the exon 1(7) GR promoter within the hippocampus of the offspring. We tested the hypothesis that NGFI-A binding to the exon 1(7) GR promoter sequence marks this sequence for histone acetylation and DNA demethylation and that such epigenetic alterations subsequently influence NGFI-A binding and GR transcription. We report that (1) NGFI-A binding to its consensus sequence is inhibited by DNA methylation, (2) NGFI-A induces the activity of exon 1(7) GR promoter in a transient reporter assay, (3) DNA methylation inhibits exon 1(7) GR promoter activity, and (4) whereas NGFI-A interaction with the methylated exon 1(7) GR promoter is reduced, NGFI-A overexpression induces histone acetylation, DNA demethylation, and activation of the exon 1(7) GR promoter in transient transfection assays. Site-directed mutagenesis assays demonstrate that NGFI-A binding to the exon 1(7) GR promoter is required for such epigenetic reprogramming. In vivo, enhanced maternal LG is associated with increased NGFI-A binding to the exon 1(7) GR promoter in the hippocampus of pups, and NGFI-A-bound exon 1(7) GR promoter is unmethylated compared with unbound exon 1(7) GR promoter. Knockdown experiments of NGFI-A in hippocampal primary cell culture show that NGFI-A is required for serotonin-induced DNA demethylation and increased exon 1(7) GR promoter expression. The data are consistent with the hypothesis that NGFI-A participates in epigenetic programming of GR expression.


Alternative splicing modulation by a LAMMER kinase impinges on developmental and transcriptome expression.
Alternative splicing is a major contributor to genome complexity, playing a significant role in various cellular functions, including signal transduction, immunity, and development. The spliceosomal machinery is responsible for the processing of nuclear RNA. Several splicing factors associated with this complex are phosphorylated by kinases that possess a conserved LAMMER motif. We demonstrate in BY-2 tobacco cells a novel role for the LAMMER motif in the maintenance of proper subnuclear localization. Furthermore, high expression of the LAMMER kinase in Arabidopsis plants modulated the alternative splicing of specific endogenous genes and resulted in abnormal plant development and a novel transcriptome profile. A prominent feature was the upregulation of genes that play a role in protein turnover, suggesting a moderating function for these gene products in the control of alternative splicing events. Together, these results demonstrate alternative splicing modulation as a result of phosphorylation activity, providing an opportunity to study its global effect on the plasticity of plant development and gene expression at the organism level.


Targeting the Glucocorticoid Receptor Reduces Binge-Like Drinking in High Drinking in the Dark (HDID-1) Mice.
Chronic alcohol exposure can alter glucocorticoid receptor (GR) function in some brain areas that promotes escalated and compulsive-like alcohol intake. GR antagonism can prevent dependence-induced escalation in drinking, but very little is known about the role of GR in regulating high-risk nondependent alcohol intake. Here, we investigate the role of GR in regulating binge-like drinking and aversive responses to alcohol in the High Drinking in the Dark (HDID-1) mice, which have been selectively bred for high blood ethanol (EtOH) concentrations (BECs) in the Drinking in the Dark (DID) test, and in their founder line, the HS/NPT. In separate experiments, male and female HDID-1 mice were administered one of several compounds that inhibited GR or its negative regulator, FKBP51 (mifepristone [12.5, 25, 50, 100 mg/kg], CORT113176 [20, 40, 80 mg/kg], and SAFit2 [10, 20, 40 mg/kg]) during a 2-day DID task. EtOH consumption and BECs were measured. EtOH conditioned taste and place aversion (CTA and CPA, respectively) were measured in separate HDID-1 mice after mifepristone administration to assess GR's role in regulating the conditioned aversive effects of EtOH. Lastly, HS/NPT mice were administered CORT113176 during DID to assess whether dissimilar effects from those of HDID-1 would be observed, which could suggest that selective breeding had altered sensitivity to the effects of GR antagonism on binge-like drinking. GR antagonism (with both mifepristone and CORT113176) selectively reduced binge-like EtOH intake and BECs in the HDID-1 mice, while inhibition of FKBP51 did not alter intake or BECs. In contrast, GR antagonism had no effect on EtOH intake or BECs in the HS/NPT mice. Although HDID-1 mice exhibit attenuated EtOH CTA, mifepristone administration did not enhance the aversive effects of EtOH in either a CTA or CPA task. These data suggest that the selection process increased sensitivity to GR antagonism on EtOH intake in the HDID-1 mice, and support a role for the GR as a genetic risk factor for high-risk alcohol intake.


Comparison of noninvasive blood pressure measurement techniques via the coccygeal artery in anesthetized cheetahs (Acinonyx jubatus).
Two indirect blood pressure measurement techniques, Doppler (DOP) sphygmomanometry and oscillometry, applied at the ventral coccygeal artery were compared with simultaneous direct blood pressure measurements at the dorsal pedal artery in 10 anesthetized, captive cheetahs (Acinonyx jubatus). The DOP method was moderately accurate, with relatively little bias (mean difference 3.8 mmHg) and 88.6% of the DOP systolic arterial pressure measurements being within 10 mmHg of the direct systolic arterial measurement. With the oscillometric (OM) method, 89.2% of the mean arterial pressure measurements were within 10 mmHg of the direct measurement and had the least bias (mean difference 2.3 mmHg), 80.7% of the systolic measurements were within 10 mmHg of the direct measurement and had the second least bias (mean difference 2.3 mmHg), and 59% of the diastolic measurements were within 10 mmHg of the direct measurement and had significant bias (mean difference 7.3 mmHg). However, DOP showed relatively poor precision (SD 11.2 mmHg) compared with OM systolic (SD 8.0 mmHg), diastolic (SD 8.6 mmHg), and mean (SD 5.7 mmHg). Both techniques showed a linear relationship with the direct technique measurements over a wide range of blood pressures. The DOP method tended to underestimate systolic measurements below 160 mmHg and overestimate systolic measurements above 160 mmHg. The OM method tended to underestimate mean pressures below 160 mm Hg, overestimate mean pressures above 160 mmHg, underestimate systolic pressures below 170 mmHg, overestimate systolic pressures above 170 mmHg, and underestimate diastolic pressures throughout the measured blood pressure range. Indirect blood pressure measurement using the ventral coccygeal artery, particularly when using an OM device for mean and systolic arterial pressure, may be useful in the clinical assessment of cheetahs when monitoring trends over time, but caution should be taken when interpreting individual values.


1H detected (13)C echo planar imaging.
Cyclic J cross polarisation (CYCLCROP) is a sensitive method for the noninvasive monitoring of (13)C distributions and fluxes. The PRAWN rotating frame Hartmann-Hahn mixing sequence ameliorates problems associated with sensitivity to Hartmann-Hahn mismatch and reduces RF power deposition. The combination of CYCLCROP with echo planar imaging (EPI) for spatial encoding of the proton detected carbon signal allows efficient use of the available signal to be made, permitting a significant improvement in the temporal resolution of any study. We report here on some initial experiments to demonstrate the feasibility of echo planar proton detected (13)C imaging using CYCLCROP based upon the PRAWN module, including the application of the technique to the measurement of transport and accumulation of (13)C-labelled sucrose in a castor bean seedling. Two methods that can be used to eliminate the effect of the J-splitting in the EP images are presented. In addition, a fast, image-based B(1) field-mapping method which may be used to quantitatively map the low frequency RF field in a dual resonant ((13)C/(1)H) probe is presented. The technique utilises the above described imaging method, permitting fully quantitative, 64x64 axial field maps to be generated in about a minute.


Fluorescence in-situ hybridization and dermoscopy in the assessment of controversial melanocytic tumors.
Although the 'gold standard' for melanoma diagnosis remains histopathological analysis, presently dermoscopists play a significant role in the diagnostic process. However, even a combined approach may not allow a clear-cut judgment on equivocal melanocytic lesions. Fluorescence in-situ hybridization (FISH) can offer assistance in the evaluation of chromosome abnormalities associated with malignancies, and its role is emerging in melanoma diagnosis. The aim of this study was to evaluate the diagnostic role of the FISH in the assessment of controversial lesions, defined as those lesions showing discrepancies between dermatoscopic and histological evaluations. Twenty clinically and histologically ambiguous melanocytic lesions were selected. After the first histopathologic diagnosis, a second pathologist examined the specimens in a blinded review for a second opinion and to identify the most suitable areas to hybridize using probes specific to RREB1, MYB, and CCND1 genes and the centromere of chromosome 6. The first histopathological evaluation led to the diagnosis of melanoma in seven cases, whereas the second identified eight cases of malignant melanoma and was in agreement with the first in 65% of cases and with dermoscopy in 40% of cases. Cytogenetic abnormalities detected by FISH are markers of malignancy that can be useful in the characterization of difficult-to-diagnose melanocytic tumors, when the dermatologist and the pathologist have a different opinions.


The Hippo pathway regulates stem cells during homeostasis and regeneration of the flatworm Macrostomum lignano.
The Hippo pathway orchestrates activity of stem cells during development and tissue regeneration and is crucial for controlling organ size. However, roles of the Hippo pathway in highly regenerative organisms, such as flatworms, are unknown. Here we show that knockdown of the Hippo pathway core genes in the flatworm Macrostomum lignano affects tissue homeostasis and causes formation of outgrowths through hyperproliferation of stem cells (neoblasts), and leads to disruption of allometric scaling during regeneration and increased size of regenerated parts. We further show that Yap, the downstream effector of the Hippo pathway, is a potential neoblast marker gene, as it is expressed in dividing cells in M. lignano and is essential for neoblast self-renewal. The phenotypes we observe in M. lignano upon knockdown of the Hippo pathway core genes and Yap are consistent with the known functions of the pathway in other model organisms and demonstrate that the Hippo pathway is functionally conserved between flatworms and mammals. This work establishes M. lignano as a productive model for investigation of the Hippo pathway.


A snapshot of Snf2 enzymes in fission yeast.
Eukaryotic chromatin is remodelled by the evolutionarily conserved Snf2 family of enzymes in an ATP-dependent manner. Several Snf2 enzymes are part of CRCs (chromatin remodelling complexes). In the present review we focus our attention on the functions of Snf2 enzymes and CRCs in fission yeast. We discuss their molecular mechanisms and roles and in regulating gene expression, DNA recombination, euchromatin and heterochromatin structure.


Volatile compounds and changes in flavour-related enzymes during cold storage of high-intensity pulsed electric field- and heat-processed tomato juices.
The effects of high-intensity pulsed electric field (HIPEF) processing (35 kV cm(-1) for 1500 micros, using 4 micros bipolar pulses at 100 Hz) on the production of volatile compounds and flavour-related enzymes in tomato juice were investigated and compared with those of thermal processing (90 degrees C for 30 or 60 s). Tomato juice treated by HIPEF showed lower residual lipoxygenase (LOX) activity (70.2%) than juice heated at 90 degrees C for 60 s (80.1%) or 30 s (93.2%). In contrast, hydroperoxide lyase (HPL) was almost completely inactivated when the juice was subjected to 90 degrees C for 60 s, whereas roughly 50% of the control tomato juice was depleted after HIPEF treatment or thermal processing at 90 degrees C for 30 s. A slight decrease was observed in the initial LOX activity of treated and untreated samples during storage, whereas initial HPL activity was strongly affected over time. HIPEF-treated juice exhibited higher levels of compounds contributing to tomato aroma than untreated and heat-treated juices throughout storage. Thus HIPEF processing can preserve flavour quality and stability of tomato juice compared with conventional thermal treatments.


Synthesizing ginsenoside Rh2 in Saccharomyces cerevisiae cell factory at high-efficiency.
Synthetic biology approach has been frequently applied to produce plant rare bioactive compounds in microbial cell factories by fermentation. However, to reach an ideal manufactural efficiency, it is necessary to optimize the microbial cell factories systemically by boosting sufficient carbon flux to the precursor synthesis and tuning the expression level and efficiency of key bioparts related to the synthetic pathway. We previously developed a yeast cell factory to produce ginsenoside Rh2 from glucose. However, the ginsenoside Rh2 yield was too low for commercialization due to the low supply of the ginsenoside aglycone protopanaxadiol (PPD) and poor performance of the key UDP-glycosyltransferase (UGT) (biopart UGTPg45) in the final step of the biosynthetic pathway. In the present study, we constructed a PPD-producing chassis via modular engineering of the mevalonic acid pathway and optimization of P450 expression levels. The new yeast chassis could produce 529.0 mg/L of PPD in shake flasks and 11.02 g/L in 10 L fed-batch fermentation. Based on this high PPD-producing chassis, we established a series of cell factories to produce ginsenoside Rh2, which we optimized by improving the C3-OH glycosylation efficiency. We increased the copy number of UGTPg45, and engineered its promoter to increase expression levels. In addition, we screened for more efficient and compatible UGT bioparts from other plant species and mutants originating from the direct evolution of UGTPg45. Combining all engineered strategies, we built a yeast cell factory with the greatest ginsenoside Rh2 production reported to date, 179.3 mg/L in shake flasks and 2.25 g/L in 10 L fed-batch fermentation. The results set up a successful example for improving yeast cell factories to produce plant rare natural products, especially the glycosylated ones.


Sclerotherapy of reticular and telangiectatic veins of the face, hands, and chest.
Prominent tortuous veins of the face and hands may result from the process of aging and constitute a source of distress for many patients. Marked telangiectases of the chest and face are similarly distressing to some patients. To determine the safety and efficacy of sclerotherapy for telangiectatic veins of the face and chest and varicose veins of the hands. Twenty facial telangiectases and tortuous veins in 14 patients and 20 total facial sites were treated with sclerotherapy. Patients did not suffer from collagen vascular diseases, had no chronic illnesses, and were thoroughly informed of the strictly cosmetic nature of the procedure. A hyperosmolar sclerosant was used in 16 of 20 sites and the remaining 4 sites were treated with a detergent sclerosant (polidocanol 0.5-0.75%). The follow-up period ranged from 9 months to 15 years. Similarly the varicose veins of 14 hands in seven patients were treated with sclerotherapy using a detergent sclerosant [sodium tetradecyl sulfate (STS)] at various concentrations (1-3%). The follow-up period ranged from 1 to 6.5 years. Tortuous veins and telangiectases on the chest of three patients were treated with either STS 0.25-0.50% or polidocanol 0.75% with a follow-up of 2-9 years. Tortuous facial veins and telangiectases had a mean improvement of 70%, with 11 of 20 sites showing a 90-100% improvement. Hand varicosities had a mean improvement of 97.8%, and all sites showed a 90-100% resolution of varicosities. Finally, the tortuous veins and telangiectases of the chest improved by 50-100% with sclerotherapy. Only minimal to moderate telangiectatic matting was observed in two patients (one in the lateral canthal area and one near the treatment site of chest reticular veins). No ulceration or hyperpigmentation occurred long term in facial or hand veins. One patient developed an arterial ulceration on the left lateral breast. Sclerotherapy of varicose and telangiectatic veins of the face, hands, and chest can be safe and effective. Care must be given to sclerotherapy in the chest area, particularly the breast.


ATP turnover and glucose dependency in hematopoietic stem/progenitor cells are increased by proliferation and differentiation.
Hematopoietic stem cells (HSCs) are quiescent cells in the bone marrow niche and are relatively dependent on glycolytic ATP production. On the other hand, differentiated cells, including hematopoietic progenitor cells (HPCs), preferentially generate ATP via oxidative phosphorylation. However, it is unclear how cellular differentiation and the cell cycle status affect nutritional requirements and ATP production in HSCs and HPCs. Using a newly developed culture system, we demonstrated that survival of HPCs was strongly dependent on glucose, whereas quiescent HSCs survived for a certain duration without glucose. Among HPCs, granulocyte/monocyte progenitors (GMPs) were particularly dependent on glucose during proliferation. By monitoring the ATP concentration in live cells, we demonstrated that the ATP level was maintained for a short duration without glucose in HSCs, possibly due to their metabolic flexibility. In addition, HSCs exhibited low ATP turnover, whereas HPCs including GMPs demonstrated high ATP turnover and required efficient ATP production from glucose. These findings show that ATP turnover and nutritional requirements differ between HSCs and HPCs according to the cell cycle and differentiation status.


Synergistic Anti-bacterial Effects of Phellinus baumii Ethyl Acetate Extracts and β-Lactam Antimicrobial Agents Against Methicillin-Resistant Staphylococcus aureus.
The development of new drugs or alternative therapies effective against methicillin-resistant Staphylococcus aureus (MRSA) is of great importance, and various natural anti-MRSA products are good candidates for combination therapies. We evaluated the antibacterial activities of a Phellinus baumii ethyl acetate extract (PBEAE) and its synergistic effects with β-lactams against MRSA. The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the PBEAE. The PBEAE synergistic effects were determined by evaluating the MICs of anti-staphylococcal antibiotic mixtures, with or without PBEAE. Anti-MRSA synergistic bactericidal effects of the PBEAE and β-lactams were assessed by time-killing assay. An ELISA was used to determine the effect of the PBEAE on penicillin binding protein (PBP)2a production. The MICs and MBCs of PBEAE against MRSA were 256-512 and 1,024-2,048 μg/mL, respectively. The PBEAE significantly reduced MICs of all β-lactams tested, including oxacillin, cefazolin, cefepime, and penicillin. However, the PBEAE had little or no effect on the activity of non-β-lactams. Time-killing assays showed that the synergistic effects of two β-lactams (oxacillin and cefazolin) with the PBEAE were bactericidal in nature (Δlog10 colony forming unit/mL at 24 hr: 2.34-2.87 and 2.10-3.04, respectively). The PBEAE induced a dose-dependent decrease in PBP2a production by MRSA, suggesting that the inhibition of PBP2a production was a major synergistic mechanism between the β-lactams and the PBEAE. PBEAE can enhance the efficacy of β-lactams for combined therapy in patients infected with MRSA.


Overexpression of hsa-miR-148a promotes cartilage production and inhibits cartilage degradation by osteoarthritic chondrocytes.
Hsa-miR-148a expression is decreased in Osteoarthritis (OA) cartilage, but its functional role in cartilage has never been studied. Therefore, our aim was to investigate the effects of overexpressing hsa-miR-148a on cartilage metabolism of OA chondrocytes. OA chondrocytes were transfected with a miRNA precursor for hsa-miR-148a or a miRNA precursor negative control. After 3, 7, 14 and 21 days, real-time PCR was performed to examine gene expression levels of aggrecan (ACAN), type I, II, and X collagen (COL1A1, COL2A1, COl10A1), matrix metallopeptidase 13 (MMP13), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) and the serpin peptidase inhibitor, clade H (heat shock protein 47), member 1 (SERPINH1). After 3 weeks, DNA content and proteoglycan and collagen content and release were determined. Type II collagen was analyzed at the protein level by Western blot. Overexpression of hsa-miR-148a had no effect on ACAN, COL1A1 and SERPINH1 gene expression, but increased COL2A1 and decreased COL10A1, MMP13 and ADAMTS5 gene expression. Luciferase reporter assay confirmed direct interaction of miR-148a and COL10A1, MMP13 and ADAMTS5. The matrix deposited by the miR-148a overexpressing cells contained more proteoglycans and collagen, in particular type II collagen. Proteoglycan and collagen release into the culture medium was inhibited, but total collagen production was increased. Overexpression of hsa-miR-148a inhibits hypertrophic differentiation and increases the production and deposition of type II collagen by OA chondrocytes, which is accompanied by an increased retention of proteoglycans. Hsa-miR-148a might be a potential disease-modifying compound in OA, as it promotes hyaline cartilage production.


14-3-3 proteins associate with phosphorylated simple epithelial keratins during cell cycle progression and act as a solubility cofactor.
14-3-3 is a ubiquitous protein family that interacts with several signal transduction kinases. We show that 14-3-3 proteins associate with keratin intermediate filament polypeptides 8 and 18 (K8/18) that are expressed in simple-type epithelia. The association is stoichiometrically significant (> or = one 14-3-3 molecule/keratin tetramer), occurs preferentially with K18, and is phosphorylation- and cell cycle-dependent in that it occurs during S/G2/M phases of the cell cycle when keratins become hyperphosphorylated. Binding of phospho-K8/18 to 14-3-3 can be reconstituted in vitro using recombinant 14-3-3 or using total cellular cytosol. Phosphatase treatment results in dissociation of 14-3-3, and dephosphorylation of phospho-K8/18 prevents reconstitution of the binding. Three cellular keratin subpopulations were analyzed that showed parallel gradients of keratin phosphorylation and 14-3-3 binding. Incubation of 14-3-3 with keratins during or after in vitro filament assembly results in sequestering of additional soluble keratin, only in cases when the keratins were hyperphosphorylated. Our results demonstrate a stoichiometrically significant cell cycle- and phosphorylation-regulated binding of 14-3-3 proteins to K18 and in vitro evidence of a simple epithelial keratin sequestering role for 14-3-3 proteins.


Bacterial Cross-Transmission between Inanimate Surfaces and Patients in Intensive Care Units under Real-World Conditions: A Repeated Cross-Sectional Study.
Background/Objectives: Contaminated surfaces play an important role in the nosocomial infection of patients in intensive care units (ICUs). This study, conducted in two ICUs at Edouard Herriot Hospital (Lyon, France), aimed to describe rooms' microbial ecology and explore the potential link between environmental contamination and patients' colonization and/or infection. Methods: Environmental samples were realized once monthly from January 2020 to December 2021 on surfaces close to the patient (bedrails, bedside table, and dedicated stethoscope) and healthcare workers' high-touch surfaces, which were distant from the patient (computer, worktop/nurse cart, washbasin, and hydro-alcoholic solution/soap dispenser). Environmental bacteria were compared to the cultures of the patients hospitalized in the sampled room over a period of ± 10 days from the environmental sampling. Results: Overall, 137 samples were collected: 90.7% of the samples close to patients, and 87.9% of the distant ones were positives. Overall, 223 bacteria were isolated, mainly: Enterococcus faecalis (15.7%), Pantoea agglomerans (8.1%), Enterobacter cloacae/asburiae (6.3%), Bacillus cereus and other Bacillus spp (6.3%), Enterococcusfaecium (5.8%), Stenotrophomonas maltophilia (5.4%), and Acinetobacter baumannii (4.9%). Throughout the study, 142 patients were included, of which, n = 67 (47.2%) were infected or colonized by at least one bacterium. In fourteen cases, the same bacterial species were found both in environment and patient samples, with the suspicion of a cross-contamination between the patient-environment (n = 10) and environment-patient (n = 4). Conclusions: In this work, we found a high level of bacterial contamination on ICU rooms' surfaces and described several cases of potential cross-contamination between environment and patients in real-world conditions.


Diet and ADHD, Reviewing the Evidence: A Systematic Review of Meta-Analyses of Double-Blind Placebo-Controlled Trials Evaluating the Efficacy of Diet Interventions on the Behavior of Children with ADHD.
Attention-deficit/hyperactivity disorder (ADHD) is a debilitating mental health problem hampering the child's development. The underlying causes include both genetic and environmental factors and may differ between individuals. The efficacy of diet treatments in ADHD was recently evaluated in three reviews, reporting divergent and confusing conclusions based on heterogeneous studies and subjects. To address this inconsistency we conducted a systematic review of meta-analyses of double-blind placebo-controlled trials evaluating the effect of diet interventions (elimination and supplementation) on ADHD. Our literature search resulted in 14 meta-analyses, six of which confined to double-blind placebo-controlled trials applying homogeneous diet interventions, i.e. artificial food color (AFC) elimination, a few-foods diet (FFD) and poly-unsaturated fatty acid (PUFA) supplementation. Effect sizes (ES) and Confidence intervals (CI) of study outcomes were depicted in a forest plot. I2 was calculated to assess heterogeneity if necessary and additional random effects subgroup meta-regression was conducted if substantial heterogeneity was present. The AFC ESs were 0.44 (95% CI: 0.16-0.72, I2 = 11%) and 0.21 (95% CI: -0.02-0.43, I2 = 68%) [parent ratings], 0.08 (95% CI: -0.07-0.24, I2 = 0%) [teacher ratings] and 0.11 (95% CI: -0.13-0.34, I2 = 12%) [observer ratings]. The FFD ESs were 0.80 (95% CI: 0.41-1.19, I2 = 61%) [parent ratings] and 0.51 (95% CI: -0.02-1.04, I2 = 72%) [other ratings], while the PUFA ESs were 0.17 (95% CI: -0.03-0.38, I2 = 38%) [parent ratings], -0.05 (95% CI: -0.27-0.18, I2 = 0%) [teacher ratings] and 0.16 (95% CI: 0.01-0.31, I2 = 0%) [parent and teacher ratings]. Three meta-analyses (two FFD and one AFC) resulted in high I2 without presenting subgroup results. The FFD meta-analyses provided sufficient data to perform subgroup analyses on intervention type, resulting in a decrease of heterogeneity to 0% (diet design) and 37.8% (challenge design). Considering the small average ESs PUFA supplementation is unlikely to provide a tangible contribution to ADHD treatment, while further research is required for AFC elimination before advising this intervention as ADHD treatment. The average FFD ES is substantial, offering treatment opportunities in subgroups of children with ADHD not responding to or too young for medication. Further FFD research should focus on establishing the underlying mechanisms of food (e.g. incrimination of gut microbiota) to simplify the FFD approach in children with ADHD.


Augmented Reality Guided Laparoscopic Surgery of the Uterus.
A major research area in Computer Assisted Intervention (CAI) is to aid laparoscopic surgery teams with Augmented Reality (AR) guidance. This involves registering data from other modalities such as MR and fusing it with the laparoscopic video in real-time, to reveal the location of hidden critical structures. We present the first system for AR guided laparoscopic surgery of the uterus. This works with pre-operative MR or CT data and monocular laparoscopes, without requiring any additional interventional hardware such as optical trackers. We present novel and robust solutions to two main sub-problems: the initial registration, which is solved using a short exploratory video, and update registration, which is solved with real-time tracking-by-detection. These problems are challenging for the uterus because it is a weakly-textured, highly mobile organ that moves independently of surrounding structures. In the broader context, our system is the first that has successfully performed markerless real-time registration and AR of a mobile human organ with monocular laparoscopes in the OR.


Cochlear metabolomics, highlighting novel insights of purine metabolic alterations in age-related hearing loss.
Aging is an inevitable phase in mammals that leads to health impairments, including hearing loss. Age-related hearing loss (AHL) leads to psychosocial problems and cognitive decline in the elderly. In this study, mean thresholds of auditory brainstem responses (ABR) and distortion-product otoacoustic emissions (DPOAE) increased at multiple frequencies in aged rats (14 months old) compared to young rats (2 months old). Using untargeted ultra-high performance liquid chromatography-mass spectroscopy (LC-MS), we quantified molecular metabolic markers in the cochlea of aged rats with hearing loss. A total of 137 different metabolites were identified in two groups, highlighting several prominent metabolic pathways related to purine metabolism; glycine, serine, and threonine metabolism; arginine and proline metabolism; and pyrimidine metabolism. In addition, the beneficial effects of purine supplementation were demonstrated in a mimetic model of senescent marginal cells (MCs). Overall, altered metabolic profiling is both the cause and manifestation of pathology, and our results suggest that cellular senescence and dysfunctional cochlear metabolism may contribute to the progression of AHL. These findings are seminal in elucidating the pathophysiological mechanisms underlying AHL and serve as a basis for future clinical predictions and interventions in AHL.


Activated protein C mediates novel lung endothelial barrier enhancement: role of sphingosine 1-phosphate receptor transactivation.
Increased endothelial cell (EC) permeability is central to the pathophysiology of inflammatory syndromes such as sepsis and acute lung injury (ALI). Activated protein C (APC), a serine protease critically involved in the regulation of coagulation and inflammatory processes, improves sepsis survival through an unknown mechanism. We hypothesized a direct effect of APC to both prevent increased EC permeability and to restore vascular integrity after edemagenic agonists. We measured changes in transendothelial electrical resistance (TER) and observed that APC produced concentration-dependent attenuation of TER reductions evoked by thrombin. We next explored known EC barrier-protective signaling pathways and observed dose-dependent APC-mediated increases in cortical myosin light chain (MLC) phosphorylation in concert with cortically distributed actin polymerization, findings highly suggestive of Rac GTPase involvement. We next determined that APC directly increases Rac1 activity, with inhibition of Rac1 activity significantly attenuating APC-mediated barrier protection to thrombin challenge. Finally, as these signaling events were similar to those evoked by the potent EC barrier-enhancing agonist, sphingosine 1-phosphate (S1P), we explored potential cross-talk between endothelial protein C receptor (EPCR) and S1P1, the receptors for APC and S1P, respectively. EPCR-blocking antibody (RCR-252) significantly attenuated both APC-mediated barrier protection and increased MLC phosphorylation. We next observed rapid, EPCR and PI 3-kinase-dependent, APC-mediated phosphorylation of S1P1 on threonine residues consistent with S1P1 receptor activation. Co-immunoprecipitation studies demonstrate an interaction between EPCR and S1P1 upon APC treatment. Targeted silencing of S1P1 expression using siRNA significantly reduced APC-mediated barrier protection against thrombin. These data suggest that novel EPCR ligation and S1P1 transactivation results in EC cytoskeletal rearrangement and barrier protection, components potentially critical to the improved survival of APC-treated patients with severe sepsis.


Dopamine transporter imaging (123)I-FP-CIT (DaTSCAN) SPET in differential diagnosis of dopa-responsive dystonia and young-onset Parkinson's disease.
Dopa-responsive dystonia (DRD) is a genetic disorder characterized by childhood onset dystonia, dominant inheritance, diurnal symptoms fluctuation and positive levodopa response. Adult-onset DRD is frequently combined with parkinsonism and can be mistaken with young onset Parkinson's disease (YOPD). Both conditions are caused by dopamine deficiency, due to nigral cells' loss in YOPD, and due to enzymatic defects in dopamine synthesis in DRD. Single photon emission tomography (SPET) with (123)I-N--fluoropropyl-2b-carbomethoxy-3b-(4-iodophenyl) nortropane ((123)I-FP-CIT)-DaTSCAN is a sensitive neuroimaging method for the assessment of nigrostriatal dopaminergic system integrity and degeneration. Our aim was to evaluate the usefulness of (123)I-FP-CIT( DaTSCAN) SPET in the differential diagnosis of DRD and YOPD in clinical practice. Brain SPET with (123)I-FP-CIT was performed in 13 patients (7 males, 6 females), age 20-58 years, with mean age of onset of their disease, 29 years, eleven patients with early onset parkinsonian symptoms and 2 with genetically proved DRD. The images were evaluated by visual and semiquantitative analyses (ROI). The ratio of specific-striatal to non specific-occipital binding was calculated. Ten out of 11 patients with YOPD had decreased accumulation of DaTSCAN in striatum, especially in putamen, that is typical findings for Parkinson's disease. In three patients DaTSCAN was normal with symmetric tracer uptake in both striata, caudate nucleus and putamen and the diagnosis of DRD was suspected. Two patients with initial dystonic symptoms and genetically proved DRD had normal DaTSCAN. In one patient after normal DaTSCAN findings the initial diagnosis of YOPD was changed to the diagnosis of DRD. Region of interest (ROI) analyses have shown significantly lower(123)I-FP-CIT binding ratios in YOPD than in DRD in all 3 regions of interest: striatum (1.95±0.32) vs (2.76±0.10) P<0.001, putamen (1.76±0.25) vs (2.84±0.14) P<0.0001 and caudate nucleus (2.37±0.51) vs (3.27±0.14) P<0.01. In conclusion, our results indicate that DaTSCAN is an objective neuroimaging method able to distinguisch neurodegenerative disease YOPD from DRD and clarify a clinical dilemma, which is important for the treatment, prognosis and genetic counseling of patients and their families.


Defect evolution behaviors from single sulfur point vacancies to line vacancies in monolayer molybdenum disulfide.
Two-dimensional monolayer transition metal dichalcogenides (TMDs) are promising candidates for many novel nanoelectronic and optoelectronic applications due to their exceptional electronic, optical, chemical and mechanical properties. Experimentally, single chalcogen point vacancies caused by electron beam irradiation are found to agglomerate into line vacancy defects in monolayer TMDs. Herein, the corresponding defect evolution behaviors from single sulfur point vacancies to line vacancies in the monolayer molybdenum disulfide (MoS2) have been systematically studied using molecular dynamics and first principles calculations. The experimental observations of the defect evolution from single sulfur point vacancies to line vacancies are reproduced at the atomic level. The results indicate that the di-vacancy line defect and a point vacancy separated by a sulfur atom in a line evolve into tri-vacancy line defects, and the di-vacancy line defects can rotate 60° clockwise or counterclockwise. Moreover, two adjacent di-vacancy line defects with an angle of 120° can evolve into tri-vacancy line defects. High temperature and large vacancy concentrations promote the defect evolution from point vacancies to line vacancies. Intriguingly, compared with the randomly distributed point vacancy defects, the line vacancy defects formed after the defect evolution significantly decrease the mechanical properties, such as the ultimate strength, ultimate strain and Young's modulus of monolayer MoS2. In addition, the mechanical properties decrease with increasing vacancy concentration and temperature for the final configurations after defect evolution in monolayer MoS2 with different vacancy concentrations at different temperatures. The band gaps of monolayer MoS2 with line vacancy defects are smaller than those with randomly distributed point vacancy defects. Therefore, our study clarifies the defect evolution behaviors from single sulfur point vacancies to line vacancies in monolayer MoS2 and opens an opportunity for the novel nanoelectronic and optoelectronic applications of monolayer TMDs.


Multiple outputation: inference for complex clustered data by averaging analyses from independent data.
This article applies a simple method for settings where one has clustered data, but statistical methods are only available for independent data. We assume the statistical method provides us with a normally distributed estimate, theta, and an estimate of its variance sigma. We randomly select a data point from each cluster and apply our statistical method to this independent data. We repeat this multiple times, and use the average of the associated theta's as our estimate. An estimate of the variance is given by the average of the sigma2's minus the sample variance of the theta's. We call this procedure multiple outputation, as all "excess" data within each cluster is thrown out multiple times. Hoffman, Sen, and Weinberg (2001, Biometrika 88, 1121-1134) introduced this approach for generalized linear models when the cluster size is related to outcome. In this article, we demonstrate the broad applicability of the approach. Applications to angular data, p-values, vector parameters, Bayesian inference, genetics data, and random cluster sizes are discussed. In addition, asymptotic normality of estimates based on all possible outputations, as well as a finite number of outputations, is proven given weak conditions. Multiple outputation provides a simple and broadly applicable method for analyzing clustered data. It is especially suited to settings where methods for clustered data are impractical, but can also be applied generally as a quick and simple tool.


Evaluation of Malaria Screening during Pregnancy with Rapid Diagnostic Tests Performed by Community Health Workers in Burkina Faso.
One of the current strategies to prevent malaria in pregnancy is intermittent preventive treatment with sulfadoxine-pyrimethamine (IPTp-SP). However, in order for pregnant women to receive an adequate number of SP doses, they should attend a health facility on a regular basis. In addition, SP resistance may decrease IPTp-SP efficacy. New or additional interventions for preventing malaria during pregnancy are therefore warranted. Because it is known that community health workers (CHWs) can diagnose and treat malaria in children, in this study screening and treatment of malaria in pregnancy by CHWs was evaluated as an addition to the regular IPTp-SP program. CHWs used rapid diagnostic tests (RDTs) for screening and artemether-lumefantrine was given in case of a positive RDT. Overall, CHWs were able to conduct RDTs with a sensitivity of 81.5% (95% confidence interval [CI] 67.9-90.2) and high specificity of 92.1% (95% CI 89.9-93.9) compared with microscopy. After a positive RDT, 79.1% of women received artemether-lumefantrine. When treatment was not given, this was largely due to the woman being already under treatment. Almost all treated women finished the full course of artemether-lumefantrine (96.4%). In conclusion, CHWs are capable of performing RDTs with high specificity and acceptable sensitivity, the latter being dependent on the limit of detection of RDTs. Furthermore, CHWs showed excellent adherence to test results and treatment guidelines, suggesting they can be deployed for screen and treat approaches of malaria in pregnancy.


Silicon-polymer hybrid slot waveguide ring-resonator modulator.
We demonstrate a ring-resonator modulator based on a silicon-polymer hybrid slot waveguide with a tunability of 12.7 pm/V at RF speeds and a bandwidth of 1 GHz, for optical wavelengths near 1550 nm. Our slot waveguides were fabricated with 193 nm optical lithography, as opposed to the electron beam lithography used for previous results. The tunability is comparable to some of the best ring-based modulators making use of the plasma dispersion effect. The speed is likely limited only by resistance in the strip-loading section, and it should be possible to realize significant improvement with improved processing.


Partitioning of macrocyclic compounds in a cationic and an anionic micellar solution: a small-angle neutron scattering study.
Following a previous investigation on partitioning of some macrocycle compounds in sodium dodecyl sulfate (SDS) and dodecyltrimethylammonium bromide (DTAB) aqueous solutions and their effect on the micellar structure, a small-angle neutron scattering (SANS) study has been performed at fixed surfactant content (0.20 mol/L) and varying macrocycle concentrations from 0.20 up to 1.0 mol/L. Conductivity measurements have been also performed in order to evaluate the effect of the presence of macrocycles on the critical micellar concentration (cmc) of the two surfactants. SANS experimental data were fitted successfully by means of a core-plus-shell monodisperse prolate ellipsoid model. It has been found that 1,4,7,10,13,16-esaoxacyclooctadecane (18C6) and 4,7,13,16-tetraoxa-1,10-diazacyclooctadecane (22) do not interact with DTAB micelles whereas their sodium complexes interact with SDS aggregates and partially localize, as a consequence of electrostatic interaction, on the micellar surface or in the Stern layer. 2,5,8,11,14,17-Hexaoxabicyclo[16.4.0] dicosane (B18C6), as a consequence of the increased hydrophobic character with respect to 18C6, interacts with DTAB hydrocarbon chains and partially localizes in the inner part of micelles. This finding has been successfully used to justify the higher amount of B18C6 compared to the 18C6 one found in the SDS micellar phase. The substituted crown ether has been found localized both on the micelle surface via complex formation and in the inner part of micelles as a consequence of the increased hydrophobic character. For all systems, the aggregate size primarily decreases with the amount of macrocycle in the micellar phase. The interpretation of cmc trends as a function ofmacrocycle concentration gives information on its distribution between micellar and aqueous phases that is in line with SANS results.


Molecular and morphological description of Ceratomyxa scophthalmi sp. nov. (Myxozoa) infecting Scophthalmus maeoticus and first report of Myxidium finnmarchicum in the Black Sea.
In the present study, 2 species of myxozoan parasites, the novel Ceratomyxa scophthalmi sp. nov. and Myxidium finnmarchicum, were documented from the gallbladder of Scophthalmus maeoticus inhabiting the Turkish Black Sea coast at Sinop. C. scophthalmi sp. nov. had mostly spherical, clavate, and fine-grained polymorphic plasmodia that ranged in diameter from 16.0 to 29.0 µm. Spores were arcuate with a convex anterior margin and concave posterior. Valves of spores were highly elongated and unequal in thickness with one valve 1.1-1.3 times thicker than the other. Valves narrowed gradually toward slightly truncated ends. The mean (range) dimensions of spores were 9.4 (8.5-10.0) µm in length and 49.6 (44.5-55.0) µm in thickness, with polar capsules that were 3.1 (2.7-3.5) µm in length and 3.2 (2.7-3.5) µm in width. Polar filaments were coiled with 4-5 turns. Large numbers of immature forms of C. scophthalmi sp. nov., which were often 'crumpled', and mature forms were found together in the gallbladder bile. Along with morphological differences, phylogenetic analysis based on 18S rDNA, including pairwise nucleotide similarities with other related Ceratomyxa species, suggests C. scophthalmi as a novel species. Moreover, this report provides the first morphological and molecular descriptions of M. finnmarchicum outside of its original geographical location and type host species.


A consensus-based gold standard for the evaluation of mass casualty triage systems.
Accuracy and effectiveness analyses of mass casualty triage systems are limited because there are no gold standard definitions for each of the triage categories. Until there is agreement on which patients should be identified by each triage category, it will be impossible to calculate sensitivity and specificity or to compare accuracy between triage systems. To develop a consensus-based, functional gold standard definition for each mass casualty triage category. National experts were recruited through the lead investigators' contacts and their suggested contacts. Key informant interviews were conducted to develop a list of potential criteria for defining each triage category. Panelists were interviewed in order of their availability until redundancy of themes was achieved. Panelists were blinded to each other's responses during the interviews. A modified Delphi survey was developed with the potential criteria identified during the interview and delivered to all recruited experts. In the early rounds, panelists could add, remove, or modify criteria. In the final rounds edits were made to the criteria until at least 80% agreement was achieved. Thirteen national and local experts were recruited to participate in the project. Six interviews were conducted. Three rounds of voting were performed, with 12 panelists participating in the first round, 12 in the second round, and 13 in the third round. After the first two rounds, the criteria were modified according to respondent suggestions. In the final round, over 90% agreement was achieved for all but one criterion. A single e-mail vote was conducted on edits to the final criterion and consensus was achieved. A consensus-based, functional gold standard definition for each mass casualty triage category was developed. These gold standard definitions can be used to evaluate the accuracy of mass casualty triage systems after an actual incident, during training, or for research.


Clinical practice guidelines for the diagnosis and management of melanoma: melanomas that lack classical clinical features.
A Cancer Council Australia multidisciplinary working group is currently revising and updating the 2008 evidence-based clinical practice guidelines for the management of cutaneous melanoma. While there have been many recent improvements in treatment options for metastatic melanoma, early diagnosis remains critical to reducing mortality from the disease. Improved awareness of the atypical presentations of this common malignancy is required to achieve this. A chapter of the new guidelines was therefore developed to aid recognition of atypical melanomas. Main recommendations: Because thick, life-threatening melanomas may lack the more classical ABCD (asymmetry, border irregularity, colour variegation, diameter > 6 mm) features of melanoma, a thorough history of the lesion with regard to change in morphology and growth over time is essential. Any lesion that is changing in morphology or growing over a period of more than one month should be excised or referred for prompt expert opinion. Changes in management as a result of the guidelines: These guidelines provide greater emphasis on improved recognition of the atypical presentations of melanoma, in particular nodular, desmoplastic and acral lentiginous subtypes, with particular awareness of hypomelanotic and amelanotic lesions.


The Role of Research in Policy and Practice: The Zika Phenomena in Puerto Rico.
On April 19, 2017, the 37th Medical Sciences Campus Annual Research and Education Forum opened with the discussion panel The role of research in policy and practice: The Zika phenomena in Puerto Rico, with Dr. Cruz M. Nazario Delgado, Dr. Carmen D. Zorrilla and Dr. Luis A. Bonilla Soto. In this article, we critically examine the public policy process of the Zika epidemic in Puerto Rico, as presented by authors Nazario and Bonilla. We argue that policymaking and the public health responses to confront the Zika epidemic in Puerto Rico took place in a political environment where different US and local actors operated to advance their goals and vision, undermining the role of knowledge, evidence and past experiences. We propose a bottom to top preventive and community empowerment approach to control the vector. This model must be built on successful policy implementation experiences with epidemics in the Island and strengthened by evidence, international guidelines and ethical principles.


Bio-inspired protein-gold nanoconstruct with core-void-shell structure: beyond a chemo drug carrier.
Chemotherapy has been widely used in clinical practice for cancer treatment. A major challenge for a successful chemotherapy is to potentiate the anticancer activity, whilst reducing the severe side effects. In this context, we design a bio-inspired protein-gold nanoconstruct (denoted as AFt-Au hereafter) with a core-void-shell structure which exhibits a high selectivity towards carcinoma cells. Anticancer drug 5-fluorouracil (5-FU) can be sequestered into the void space of the construct to produce an integrated nanoscale hybrid AFt-AuFU that exhibits an increased cellular uptake of 5-FU. More importantly, AFt-Au, serving as a bio-nano-chemosensitizer, renders carcinoma cells more susceptible to 5-FU by cell-cycle regulation, and thus, leads to a dramatic decrease of the IC50 value (i.e. the drug concentration required to kill 50% of the cell population) of 5-FU in HepG2 cells from 138.3 μM to 9.2 μM. Besides HepG2 cells, a remarkably enhanced anticancer efficacy and potentially reduced side effects are also achieved in other cell lines. Our further work reveals that the drug 5-FU is internalized into cells with AFt-Au primarily via receptor-mediated endocytosis (RME). After internalization, AFt-AuFU colocalizes with lysosomes which trigger the release of 5-FU under acidic conditions. Overall, our approach provides a novel procedure in nanoscience that promises an optimal chemotherapeutic outcome.


Motivation Modulates Brain Networks in Response to Faces Varying in Race and Status: A Multivariate Approach.
Previous behavioral and neuroimaging work indicates that individuals who are externally motivated to respond without racial prejudice tend not to spontaneously regulate their prejudice and prefer to focus on nonracial attributes when evaluating others. This fMRI multivariate analysis used partial least squares analysis to examine the distributed neural processing of race and a relevant but ostensibly nonracial attribute (i.e., socioeconomic status) as a function of the perceiver's external motivation. Sixty-one white male participants (Homo sapiens) privately formed impressions of black and white male faces ascribed with high or low status. Across all conditions, greater external motivation was associated with reduced coactivation of brain regions believed to support emotion regulation (rostral anterior cingulate cortex), introspection (middle cingulate), and social cognition (temporal pole, medial prefrontal cortex). The reduced involvement of this network irrespective of target race and status suggests that external motivation is related to the participant's overall approach to impression formation in an interracial context. The findings highlight the importance of examining network coactivation in understanding the role of external motivation in impression formation, among other interracial social processes.


The Significance of Concentration-dependent Components in Computational Models of C-Fibers.
Computational models of neurons are valuable tools that allow researchers to form and evaluate hypotheses and minimize high-cost animal work. We soon plan to use computational modeling to explore the response of different sensory fiber types to long duration external stimulation to try to selectively block nociceptive C-fibers. In this work, we modified an existing C-fiber-specific axon model to additionally include concentration-dependent conductance changes, the contribution of longitudinal current flow to changes in local concentrations, and longitudinal currents generated by concentration gradients along the axon. Then, we examined the impact of these additional elements on the modeled action potential properties, activity-dependent latency increases, and concentration changes due to external stimulation. We found that these additional model elements did not significantly affect the action potential properties or activity-dependent behavior, but they did have a significant impact on the modeled response to external long duration stimulation.Clinical Relevance- This presents a computational model that can be used to help investigate and develop electrical stimulation therapies for pathological pain.


MELK promotes HCC carcinogenesis through modulating cuproptosis-related gene DLAT-mediated mitochondrial function.
Cuproptosis caused by copper overload is mediated by a novel regulatory mechanism that differs from previously documented mechanisms regulating cell death. Cells dependent on mitochondrial respiration showed increased sensitivity to a copper ionophore elesclomol that induced cuproptosis. Maternal embryonic leucine zipper kinase(MELK) promotes tumorigenesis and tumor progression through the PI3K/mTOR pathway, which exerts its effects partly by targeting the pyruvate dehydrogenase complex(PDHc) and reprogramming the morphology and function of mitochondria. However, the role of MELK in cuproptosis remains unclear. Here, we validated that elevated MELK expression enhanced the activity of PI3K/mTOR signaling and subsequently promoted Dihydrolipoamide S-Acetyltransferase (DLAT) expression and stabilized mitochondrial function. This regulatory effect helped to improve mitochondrial respiration, eliminate excessive intracellular reactive oxygen species (ROS), reduce intracellular oxidative stress/damage and the possibility of mitochondria-induced cell fate alternations, and ultimately promote the progression of HCC. Meanwhile, elesclomol reduced translocase of outer mitochondrial membrane 20(TOM 20) expression and increased DLAT oligomers. Moreover, the above changes of MELK to HCC were abolished by elesclomol. In conclusion, MELK enhanced the levels of the cuproptosis-related signature(CRS) gene DLAT (especially the proportion of DLAT monomer) by activating the PI3K/mTOR pathway, thereby promoting elesclomol drug resistance, altering mitochondrial function, and ultimately promoting HCC progression.


Platelet function in stored heparinised autologous blood is not superior to in patient platelet function during routine cardiopulmonary bypass.
In cardiac surgery, cardiopulmonary bypass (CPB) and unfractionated heparin have negative effects on blood platelet function. In acute normovolemic haemodilution autologous unfractionated heparinised blood is stored ex-vivo and retransfused at the end of the procedure to reduce (allogeneic) transfusion requirements. In this observational study we assessed whether platelet function is better preserved in ex vivo stored autologous blood compared to platelet function in the patient during CPB. We measured platelet aggregation responses pre-CPB, 5 min after the start of CPB, at the end of CPB, and after unfractionated heparin reversal, using multiple electrode aggregometry (Multiplate®) with adenosine diphosphate (ADP), thrombin receptor activating peptide (TRAP) and ristocetin activated test cells. We compared blood samples taken from the patient with samples taken from 100 ml ex-vivo stored blood, which we took to mimick blood storage during normovolemic haemodilution. Platelet function declined both in ex-vivo stored blood as well as in blood taken from the patient. At the end of CPB there were no differences in platelet aggregation responses between samples from the ex vivo stored blood and the patient. Ex vivo preservation of autologous blood in unfractionated heparin does not seem to be profitable to preserve platelet function.


Resonant Doppler flow imaging and optical vivisection of retinal blood vessels.
For Fourier domain optical coherence tomography any sample movement during camera integration causes blurring of interference fringes and as such reduction of sensitivity for flow detection. The proposed method overcomes this problem by phase-matching a reference signal to the sample motion. The interference fringes corresponding to flow signal will appear frozen across the detector whereas those of static sample structures will be blurred resulting in enhanced contrast for blood vessels. An electro-optic phase modulator in the reference arm, driven with specific phase cycles locked to the detection frequency, allows not only for qualitative but also for quantitative flow detection already from the relative signal intensities. First applications to extract in-vivo retinal flow and to visualize 3D vascularization, i.e. optical vivisection, are presented.


A multiparameter analysis of sickle erythrocytes in patients undergoing hydroxyurea therapy.
During 24 weeks of hydroxyurea treatment, we monitored red blood cell (RBC) parameters in three patients with sickle cell disease, including F-cell and F-reticulocyte profiles, distributions of delay times for intracellular polymerization, sickle erythrocyte adherence to human umbilical vein endothelial cells in a laminar flow chamber, RBC phthalate density profiles, mean corpuscular hemoglobin concentration and cation content, reticulocyte mean corpuscular hemoglobin concentration, 1H-nuclear magnetic resonance transverse relaxation rates of packed RBCs, and plasma membrane lateral and rotational mobilities of band 3 and glycophorins. Hydroxyurea increases the fraction of cells with sufficiently long delay times to escape the microcirculation before polymerization begins. Furthermore, high pretreatment adherence to human umbilical vein endothelial cells of sickle RBCs decreased to normal after only 2 weeks of hydroxyurea treatment, preceding the increase in fetal hemoglobin levels. The lower adhesion of sickle RBCs to endothelium would facilitate escape from the microcirculation before polymerization begins. Hydroxyurea shifted several biochemical and biophysical parameters of sickle erythrocytes toward values observed with hemoglobin SC disease, suggesting that hydroxyurea moderates sickle cell disease toward the milder, but still clinically significant, hemoglobin SC disease. The 50% reduction in sickle crises documented in the Multicenter Study of Hydroxyurea in Sickle Cell Disease is consistent with this degree of erythrocyte improvement.


Subversion of monocyte functions by coxiella burnetii: impairment of the cross-talk between alphavbeta3 integrin and CR3.
Several intracellular pathogens exploit macrophages as a niche for survival and replication. The success of this strategy requires the subversion or the avoidance of microbicidal functions of macrophages. Coxiella burnetii, the agent of Q fever, is a strictly intracellular bacterium that multiplies in myeloid cells. The survival of C. burnetii may depend on the selective use of macrophage receptors. Virulent C. burnetii organisms were poorly internalized but survived successfully in human monocytes, whereas avirulent variants were efficiently phagocytosed but were also rapidly eliminated. The uptake of avirulent organisms was mediated by leukocyte response integrin (alphavbeta3 integrin) and CR3 (alphaMbeta2 integrin), as demonstrated by using specific Abs and RGD sequence-containing peptides. The phagocytic efficiency of CR3 depends on its activation via alphavbeta3 integrin and integrin-associated protein. Indeed, CR3-mediated phagocytosis of avirulent C. burnetii was abrogated in macrophages from integrin-associated protein-/- mice. In contrast, the internalization of virulent C. burnetii organisms involved the engagement of alphavbeta3 integrin but not that of CR3. The pretreatment of monocytes with virulent C. burnetii organisms prevented the CR3-mediated phagocytosis of zymosan particles and CR3 activation assessed by the expression of the 24 neo-epitope. We conclude that the virulence of C. burnetii is associated with the engagement of alphavbeta3 integrin and the impairment of CR3 activity, which probably results from uncoupling alphavbeta3 integrin from integrin-associated protein. This study describes a strategy not previously reported of phagocytosis modulation by intracellular pathogens.


Rosacea fulminans: two case reports and review of the literature.
Rosacea fulminans is a rare and severe inflammatory dermatosis which affects predominantly childbearing women. It is characterized by sudden onset and it usually localizes exclusively on the centrofacial areas, presenting with numerous fluctuant inflammatory nodules and papules which may coalesce. Treatment with isotretinoin in combination with topical and systemic corticosteroids is successful. Clearance of lesions may be obtained under systemic treatment with no or minimal scarring outcomes. Due to rare incidence its pathophysiological mechanisms, diagnosis and management remain controversial. We report two cases of rosacea fulminans arisen in otherwise healthy people and completely healed after treatment. Our aim is to share our experience about this disease in order to increase knowledge about its diagnosis, management and its treatment. We also make a review of the literature of this peculiar dermatosis.


Secretion of biologically active human interleukin-2 and interleukin-4 from genetically modified tobacco cells in suspension culture.
Biologically active human interleukin-2 (IL-2) and IL-4, key lymphokines involved in immune regulation, were produced and secreted into the medium by genetically modified Nicotiana tabacum cells grown in suspension culture. Secretion through the plasma membrane and cell wall into the medium was facilitated by the natural mammalian leader sequences. IL-2 and IL-4 were detected in the medium at concentrations of 0.10 and 0.18 microgram/mL, respectively, although higher levels were detected within the lymphokine-producing cells (approximately 0.80 microgram/mL for IL-2 and approximately 0.28 microgram/mL for IL-4). By Western blot, IL-4 was found to be secreted as two small polypeptides with molecular masses of approximately 18-20 kDa. The biological activity of IL-2 was determined by cell proliferation of the IL-2-dependent murine CTLL-2 cell line, while that of IL-4 was determined by cell proliferation of the CTLL-2 cell line [CT.h4S] which was stably transfected with the human IL-4 receptor. These findings indicate that plant suspension culture can be used to produce and secrete into the medium a variety of biologically active mammalian proteins that are of clinical and diagnostic relevance.


Can You Hear What I Think? Theory of Mind in Young Children With Moderate Hearing Loss.
The first aim of this study was to examine various aspects of Theory of Mind (ToM) development in young children with moderate hearing loss (MHL) compared with hearing peers. The second aim was to examine the relation between language abilities and ToM in both groups. The third aim was to compare the sequence of ToM development between children with MHL and hearing peers. Forty-four children between 3 and 5 years old with MHL (35 to 70 dB HL) who preferred to use spoken language were identified from a nationwide study on hearing loss in young children. These children were compared with 101 hearing peers. Children were observed during several tasks to measure intention understanding, the acknowledgement of the other's desires, and belief understanding. Parents completed two scales of the child development inventory to assess expressive language and language comprehension in all participants. Objective language test scores were available from the medical files of children with MHL. Children with MHL showed comparable levels of intention understanding but lower levels of both desire and belief understanding than hearing peers. Parents reported lower language abilities in children with MHL compared with hearing peers. Yet, the language levels of children with MHL were within the average range compared with test normative samples. A stronger relation between language and ToM was found in the hearing children than in children with MHL. The expected developmental sequence of ToM skills was divergent in approximately one-fourth of children with MHL, when compared with hearing children. Children with MHL have more difficulty in their ToM reasoning than hearing peers, despite the fact that their language abilities lie within the average range compared with test normative samples.


Epidermal growth factor receptor activation differentially regulates claudin expression and enhances transepithelial resistance in Madin-Darby canine kidney cells.
Tight junctions (TJs) are the most apical cell-cell junctions, and claudins, the recently identified TJ proteins, are critical for maintaining cell-cell adhesion in epithelial cell sheets. Based on their in vivo distribution and the results of overexpression studies, certain claudins, including claudin-1 and -4, are postulated to increase, whereas other claudins, especially claudin-2, are postulated to decrease the overall transcellular resistance. The overall ratio among claudins expressed in a cell/tissue has been hypothesized to define the complexity of TJs. Disruption of the TJs contributes to various human diseases, and a correlation between reduction of TJ function and tumor dedifferentiation has been postulated. The epidermal growth factor (EGF) receptor (EGFR) is overexpressed in a wide spectrum of epithelial cancers, and its expression correlates with a more metastatic cancer phenotype. However, normal functioning of EGFR is essential for normal epithelial cell proliferation and differentiation. The role of EGFR-dependent signaling in the development and maintenance of epithelial TJ integrity has not been studied in detail. This study demonstrates that, in polarized Madin-Darby canine kidney II cells, EGF-induced EGFR activation significantly inhibited claudin-2 expression while simultaneously inducing cellular redistribution and increased expression of claudin-1, -3, and -4. Accompanying these EGF-induced changes in claudin expression was a 3-fold increase in transepithelial resistance, a functional measure of TJs. In contrast, there were no alterations in protein expression and/or intracellular localization of other TJ-related proteins (ZO-1 and occludin) or adherens junction-associated proteins (E-cadherin and beta-catenin), suggesting that EGF regulates TJ function through selective and differential regulation of claudins.


Tumoral Melanosis in the Setting of Targeted Immunotherapy for Metastatic Melanoma-A Single Institutional Experience and Literature Review.
Tumoral melanosis clinically resembles metastatic melanoma, occurs in the context of regressed disease, and requires evaluation to rule out underlying melanoma and metastatic disease. Histopathology demonstrates a nodular infiltrate of melanophages in the dermis, subcutaneous tissue, deep soft tissue, or lymph nodes in the absence of viable melanocytes. Recent limited reports of tumoral melanosis in the context of immunotherapy with ipilimumab (monoclonal antibody targeting CTLA-4) as well as nivolumab and pembrolizumab (humanized monoclonal antibodies against programmed death 1 receptor) highlight a unique presentation representative of treatment-related tumor regression and an association with a favorable clinical response. To describe our experience with tumoral melanosis in the setting of immunotherapy for metastatic melanoma and elucidate the clinical and histopathological features. Retrospective case series from a single tertiary care institution. We describe 10 cases of patients with metastatic melanoma who received treatment with immunotherapy before the development of tumoral melanosis. Length of time between the initiation of therapy and the onset of tumoral melanosis ranged from 2 to 20 months with a mean time of 10 months. At the end of the follow-up period, 8 patients were classified as having a complete or partial response to treatment with immunotherapy. One patient had progression of visceral and cutaneous disease on ipilimumab despite developing tumoral melanosis, and 1 patient had yet to undergo repeat imaging. Furthermore, at the end of follow-up, 3 patients were alive with no evidence of active disease, 5 patients were alive with disease, and 1 patient was deceased, although this patient died of a cardiovascular event unrelated to his underlying melanoma. Of the patients who were classified as alive with disease, 2 patients had minimal remaining disease, and 2 patients had an almost complete response on immunotherapy with recurrence of visceral metastases after immunotherapy was discontinued. One patient developed new peritoneal and cutaneous metastases on pembrolizumab despite development of tumoral melanosis. The underlying biologic mechanisms and prognostic implications of tumoral melanosis in the setting of immunotherapy remain to be elucidated. Further prospective studies with a larger cohort and prolonged follow-up are necessary to better understand the incidence, prevalence, and oncologic outcomes in patients with tumoral melanosis who receive immunotherapy.


Seminal vesicles and diabetic neuropathy: ultrasound evaluation.
The aim of the study was to evaluate the ultrasound characteristics of the seminal vesicles (SV) of infertile patients with diabetes and neuropathy, and possible changes in relation to duration of diabetes. Sixty infertile patients with type 2 diabetes and symptomatic neuropathy were selected. Patients were divided into 3 groups according to duration of diabetes (group A ≤ 5 years, group B between 5 and 15 years, and group C ≥ 15 years). A pathological control group of 20 infertile patients without diabetes and a real control group of 20 healthy fertile men were selected and compared. Patients underwent prostate-vesicular transrectal ultrasonography and sperm analysis. The following ultrasound parameters were recorded: 1) body anteroposterior diameter (APD); 2) fundus APD; 3) parietal thickness of the right and left SVs; 4) number of polycyclic areas within both SVs; 5) fundus-to-body ratio; 6) difference of the parietal thickness between the right and the left SVs; and 7) pre-ejaculatory and postejaculatory APD difference. All patients with diabetes had a significantly (P < .05) higher fundus-to-body ratio compared with controls. Group C had a significantly (P < .05) higher fundus-to-body ratio compared with other diabetes groups. There was no significant difference (P > .05) relative to the number of polycyclic areas in patients with diabetes and controls. All patients with diabetes had a significantly lower (P < .05) preejaculatory and postejaculatory difference in body SV APD compared with controls. Group A and group B had a similar preejaculatory and postejaculatory difference in body SV APD, whereas this difference was significantly (P < .05) lower in group C. In conclusion, infertile patients with diabetes and neuropathy have peculiar SV ultrasound features suggestive of functional atony, and duration of disease is associated with worse changes in ultrasound findings.


Acute otitis media: management and surveillance in an era of pneumococcal resistance. Drug-Resistant Streptococcus Pneumoniae Therapeutic Working Group.
Experts in the management of otitis media and the Drug-resistant Streptococcus pneumoniae Therapeutic Working Group were convened by the Centers for Disease Control and Prevention to respond to changes in antimicrobial susceptibility among pneumococci. The objective was to provide consensus recommendations for the management of acute otitis media (AOM) and for the surveillance of drug-resistant Streptococcus pneumoniae. After summarizing published and unpublished data from the scientific literature and the experience of the panel members, the group concluded that oral amoxicillin should remain the first-line antimicrobial agent for treating AOM. For patients with clinically defined treatment failure after 3 days of therapy, useful alternative agents include amoxicillin-clavulanate, cefuroxime axetil, and intramuscular ceftriaxone. The group also made recommendations to improve surveillance and to obtain antimicrobial susceptibility patterns for local geographic areas.


Modelling the potential long-term survival benefit of evinacumab treatment vs. standard of care in patients with homozygous familial hypercholesterolaemia.
Despite intensive lipid-lowering therapies (LLTs), most patients with homozygous familial hypercholesterolaemia (HoFH) do not achieve guideline recommended low-density lipoprotein cholesterol (LDL-C) targets and are at increased risk of premature cardiovascular death. This analysis aimed to predict the impact of evinacumab and standard-of-care LLTs on life expectancy in an HoFH population using mathematical modelling. Mathematical models were developed using efficacy data for evinacumab from the phase 3 ELIPSE HoFH trial plus efficacy data for standard-of-care LLTs from peer-reviewed publications. Treatment strategies evaluated included (i) untreated, (ii) high-intensity statin (HIS) only, (iii) HIS plus ezetimibe, (iv) HIS plus ezetimibe plus proprotein convertase subtilisin/kexin type 9 inhibitor (PCSK9i), and (v) HIS plus ezetimibe plus PCSK9i plus evinacumab. Markov analyses were used to assess differences in survival probability for different LLT strategies. The median survival for untreated HoFH patients was only 33-43 years, depending on different assumptions on baseline untreated LDL-C levels. In the most robust model, we estimated that HIS increased median survival by 9 years and ezetimibe further increased median survival by an additional 9 years. When PCSK9i was added on top of HIS plus ezetimibe, median survival was further improved by 14 years. Finally, the addition of evinacumab to standard-of-care LLTs was estimated to increase median survival by ∼12 years. In this mathematical modelling analysis, evinacumab treatment could potentially increase long-term survival vs. standard-of-care LLTs for patients with HoFH.


Evaluation of biological monitoring markers using genomic and proteomic analysis for automobile emission inspectors and waste incinerating workers exposed to polycyclic aromatic hydrocarbons or 2,3,7,8,-tetracholrodedibenzo-p-dioxins.
In this study, we investigated the effects of PAHs and dioxin on mRNA and plasma protein expression using genomic and proteomic analysis for automobile emission inspectors and waste incineration workers. About 54 workers from automobile emission inspection offices, 31 workers from waste incinerating company and 84 unexposed healthy subjects were enrolled in this study. Urine and air samples were collected and analyzed by HPLC and GC/MS. Comet assays were carried out to evaluate any DNA damage in mononuclear and polynuclear cells. A significant difference in Olive tail moments in mononuclear cells was observed between exposed and control subjects (P < 0.0001). To examine the differences of the gene expression profile in automobile emission inspectors and waste incineration workers, radioactive complementary DNA microarrays were used to evaluate changes in the expression of 1,152 total genes. The gene expression profiles showed that 11 genes were up-regulated and 4 genes were down-regulated in waste incinerating workers as compared with controls. Plasma proteins were analyzed by 2-dimentional electrophoresis with pH 3-10 NL IPG Dry strip. The protein expression profiles showed that 8 proteins were up- regulated and 1 protein, haptoglobin, was down- regulated in automobile emission inspectors and waste incineration workers. Serum paraoxonase/ arylesterase was found only in the plasma of waste incineration workers. The expression of genes and proteins involved in oxidative stress were up-regulated in both automobile emission inspectors and waste incineration workers. Several proteins, such as transthyrethin, sarcolectin and haptoglobin, that were highly up- or down-regulated, could serve as biological monitoring markers for future study.


Creating "an oasis of hope, inclusion and connection": students and stakeholders' experiences of a pilot Recovery College.
Recovery Colleges are an innovative approach to promoting personal recovery for people experiencing mental illness. This study was to explore experiences of students, supporters, staff, educators and external stakeholders (i.e. partner organisations) of a pilot Recovery College in the Australian Capital Territory (ACTRC), and the impact of participation in the College for students and supporters. ACTRC students, supporters, staff and educators, and external stakeholders were invited to participate in a mixed-method evaluation via an online survey, interviews and/or focus groups. The survey included questions regarding experiences and recovery-orientation of the College environment, and for students and supporters only, satisfaction with the College. Qualitative data from interviews and focus groups was inductively coded, thematically analysed and triangulated with survey responses. The findings suggest that the ACTRC provides a safe space, promotes meaningful connections within and beyond the college, and offers steppingstones supporting recovery and growth. Participants spoke positively about cross institutional partnerships and collaboration with several organisations within the ACT. This evaluation reiterates the role of Recovery Colleges as an innovative approach to promoting personal recovery for people living with mental illness. Adequate resourcing and collaboration are essential in realising the value of co-production whilst ensuring sustainability.


Few fathers-to-be prefer caesarean section for the birth of their baby.
The objective of this study was to investigate prospective fathers' preferences for caesarean section and associated factors. Data were collected by means of a questionnaire given in mid-pregnancy to 1105 fathers-to-be in northern Sweden. In total, 6.4% of fathers preferred a caesarean section. The factors associated with a preference for caesarean section were a wish to plan the date of the baby's birth [prevalence ratio (PR) 6.0], a previous negative birth experience (PR 8.6) and previous experience of a caesarean section (PR 5.7).


Optimizing Engagement in an mHealth Intervention for Diabetes Support During Pregnancy: the Role of Baseline Patient Health and Behavioral Characteristics.
Mobile health (mHealth) technology that addresses diabetes mellitus in pregnancy has the potential to improve maternal and child health while diminishing socioeconomic and racial disparities. Little is known about health literacy, electronic health literacy, or patient characteristics that contribute to increased mHealth use. In this pilot study, we aimed to examine patient factors associated with user engagement with a novel app for diabetes support during pregnancy. Low-income pregnant individuals with gestational or type 2 diabetes mellitus were recruited for a 2-week usability assessment of a novel mHealth smartphone application, "SweetMama," designed to improve diabetes care. This analysis of user characteristics was a pilot assessment of data from the usability study phase. Participants completed assessments of patient health literacy, electronic health literacy, diabetes self-efficacy, and activation. User metrics (eg, sessions and total duration of use) were evaluated by participant characteristics. Descriptive analyses were conducted to assess outcomes with respect to patient's clinical history, health literacy, electronic health literacy, diabetes self-efficacy, and activation. Twenty-two diverse, low-income pregnant patients with either gestational or type 2 diabetes engaged with SweetMama for 14 days. Participants with gestational diabetes had greater mean minutes of use per session and mean total duration of use than those with type 2 diabetes. Participants with greater electronic health literacy, lower patient activation, and greater self-efficacy demonstrated greater total duration of use. No user metrics differed significantly by participant general health literacy. Findings of this pilot study suggest engagement with a novel mHealth app for diabetes support during pregnancy varied by baseline behavioral and clinical characteristics. Results could indicate that digital health literacy better differentiates engagement patterns than general health literacy.


Early heart rate responses to standardized trauma-related pictures predict posttraumatic stress disorder: a prospective study.
To investigate whether chronic posttraumatic stress disorder can be predicted by heart rate responses (HRR) and skin conductance responses (SCR) to standardized trauma-related pictures at 1 month after trauma has ocurred. Trauma survivors with PTSD report heightened physiological responses to a wide range of stimuli. It has been suggested that associative learning and stimulus generalization play a key role in the development of these symptoms. Some studies have found that trauma survivors with PTSD show greater physiological responses to individualized trauma reminders in the initial weeks after trauma than those without PTSD. Survivors of motor vehicle accidents or physical assaults (n = 166) watched standardized trauma-related, generally threatening, and neutral pictures at 1 month post trauma, as their HRR and SCR were recorded. PTSD symptoms were assessed with structured clinical interviews at 1 month and 6 months; self-reports of fear responses and dissociation during trauma were obtained soon after the trauma. At 1 month, trauma survivors with PTSD showed greater HRR to trauma-related pictures than those without PTSD, but not to general threat or neutral pictures. HRR to trauma-related pictures predicted PTSD severity at 1 month and 6 months, and were related to fear and dissociation during trauma. SCR were not related to PTSD. HRR to standardized trauma reminders at 1 month after the trauma differentiate between trauma survivors with and without PTSD, and predict chronic PTSD. RESULTS are consistent with a role of associative learning in PTSD and suggest that early stimulus generalization may be an indicator of risk for chronic PTSD.


A Species-Wide Inventory of NLR Genes and Alleles in Arabidopsis thaliana.
Infectious disease is both a major force of selection in nature and a prime cause of yield loss in agriculture. In plants, disease resistance is often conferred by nucleotide-binding leucine-rich repeat (NLR) proteins, intracellular immune receptors that recognize pathogen proteins and their effects on the host. Consistent with extensive balancing and positive selection, NLRs are encoded by one of the most variable gene families in plants, but the true extent of intraspecific NLR diversity has been unclear. Here, we define a nearly complete species-wide pan-NLRome in Arabidopsis thaliana based on sequence enrichment and long-read sequencing. The pan-NLRome largely saturates with approximately 40 well-chosen wild strains, with half of the pan-NLRome being present in most accessions. We chart NLR architectural diversity, identify new architectures, and quantify selective forces that act on specific NLRs and NLR domains. Our study provides a blueprint for defining pan-NLRomes.


Addressing Puberty Challenges for Adolescents With Autism Spectrum Disorder: A Survey of Occupational Therapy Practice Trends.
Understanding occupational therapy practice patterns for adolescents who have autism spectrum disorder (ASD) provides a baseline to determine areas of need and effective interventions. To identify puberty-related challenges addressed and interventions used by occupational therapy practitioners with adolescents ages 8-16 yr with ASD. A nonexperimental survey design with a stratified random sample was used. The Occupational Therapy-Autism and Puberty Survey was distributed in the United States online through the Qualtrics platform (available October 2017-January 2018). Occupational therapists (89%) or occupational therapy assistants with experience with adolescents who have ASD were included. The 71 eligible respondents were primarily White women with an average 16 yr of experience. Outcomes and Measures: Nine key challenges and 11 interventions regarding puberty and ASD were identified. Hypotheses were generated before data collection. More practitioners (>47%) addressed emotional regulation and personal hygiene, and fewer (<26%) addressed menstruation, safety, and masturbation. Nearly all practitioners (>92%) reported using social learning approaches and behavioral strategies, and most (>80%) reporting having received education in these interventions. Significant differences were found between reported education and use of behavioral skills training incorporating applied behavioral analysis (p < .001), parent training (p = .002), and technology (p = .003). Practitioners are neglecting to address puberty-specific challenges with adolescents who have ASD. Opportunities exist to expand the interventions used and challenges addressed by occupational therapy practitioners for adolescents with ASD. What This Article Adds: This article expands occupational therapy literature by giving a baseline understanding of the puberty-related challenges that practitioners are addressing with adolescents who have ASD. This information can assist the profession in developing effective interventions and educational opportunities to allow practitioners to successfully address puberty-specific challenges.


Influence of the magnetic state on the chemical order-disorder transition temperature in Fe-Ni permalloy.
In magnetic alloys, the effect of finite temperature magnetic excitations on phase stability below the Curie temperature is poorly investigated, although many systems undergo phase transitions in this temperature range. We consider random Ni-rich Fe-Ni alloys, which undergo chemical order-disorder transition approximately 100 K below their Curie temperature, to demonstrate from ab initio calculations that deviations of the global magnetic state from ideal ferromagnetic order due to temperature induced magnetization reduction have a crucial effect on the chemical transition temperature. We propose a scheme where the magnetic state is described by partially disordered local magnetic moments, which in combination with Heisenberg Monte Carlo simulations of the magnetization allows us to reproduce the transition temperature in good agreement with experimental data.


Stroke prevention in atrial fibrillation patients with chronic kidney disease.
Chronic kidney disease (CKD) is prevalent in elderly patients with atrial fibrillation and is an independent risk factor for stroke. Warfarin anticoagulation is efficacious for stroke prevention in atrial fibrillation patients with moderate CKD (stage III, estimated glomerular filtration rate 30-59 mL/min), but recent observational studies have challenged its value for patients with end-stage renal disease requiring dialysis. The novel oral anticoagulants (i.e., dabigatran, apixaban, rivaroxaban) all undergo renal metabolism to varying degrees, and hence dosing, efficacy, and safety require special consideration in CKD patients. In randomized trials to date involving 11,169 patients with moderate CKD, the novel oral anticoagulants performed well, with similar efficacy and safety profiles as for non-CKD patients. For atrial fibrillation patients with stage III CKD, the available data are strongest for dabigatran 150 mg twice daily as superior to warfarin for stroke prevention and for apixaban as superior to warfarin regarding reduced major hemorrhage. Renal function should be monitored at least annually in patients receiving a novel oral anticoagulant, and more often in elderly patients and those with underlying CKD or comorbidities who are at special risk for dehydration and deterioration of renal function. Much remains to be learned about the optimal use of the novel oral anticoagulants in CKD patients; additional studies about optimal dosing of the novel oral anticoagulants and frequency of monitoring renal function in CKD patients with atrial fibrillation are needed. Anticoagulation options for hemodialysis patients require testing in randomized trials.


A neurobiological model for the effects of early brainstem functioning on the development of behavior and emotion regulation in infants: implications for prenatal and perinatal risk.
Neurobiological models propose an evolutionary, vertical-integrative perspective on emotion and behavior regulation, which postulates that regulatory functions are processed along three core brain systems: the brainstem, limbic, and cortical systems. To date, few developmental studies applied these models to research on prenatal and perinatal risk. We propose a conceptual model that incorporates three integrated levels of observations for the study of early risk: (a) brainstem-related physiological regulation of cyclic processes and sensory integration, e.g., vagal regulation, circadian rhythms; (b) emotion and attention regulation capacities that draw on the integration of brainstem and limbic systems; and (c) higher-level outcomes that draw on the intactness of brainstem and limbic networks, including socio-emotional self-regulation, inhibitory control, and cognitive processing. We discuss implications of the model for the development of regulatory capacities during the prenatal and early postnatal stages in infants born with specific perinatal risk. We underscore the importance of assessing sub-cortical and brainstem systems and the longitudinal effects of transitory brainstem dysfunction on physiological homeostasis, motivation, arousal-modulated attention, stress reactivity, and mother-infant co-regulation. The assessment of brainstem dysfunction can be conducted during hospitalization and may help detect infants at risk for the development of self-regulatory deficits at the first weeks of life.


Behind the Leaves: Estimation of Occluded Grapevine Berries With Conditional Generative Adversarial Networks.
The need for accurate yield estimates for viticulture is becoming more important due to increasing competition in the wine market worldwide. One of the most promising methods to estimate the harvest is berry counting, as it can be approached non-destructively, and its process can be automated. In this article, we present a method that addresses the challenge of occluded berries with leaves to obtain a more accurate estimate of the number of berries that will enable a better estimate of the harvest. We use generative adversarial networks, a deep learning-based approach that generates a highly probable scenario behind the leaves exploiting learned patterns from images with non-occluded berries. Our experiments show that the estimate of the number of berries after applying our method is closer to the manually counted reference. In contrast to applying a factor to the berry count, our approach better adapts to local conditions by directly involving the appearance of the visible berries. Furthermore, we show that our approach can identify which areas in the image should be changed by adding new berries without explicitly requiring information about hidden areas.


Molecular characterization of a saline-soluble lectin from a parasitic fungus. Extensive sequence similarities between fungal lectins.
It has been proposed that the interactions between several parasitic and pathogenic fungi and their hosts are mediated by soluble lectins present in the fungus. We have cloned and analyzed a gene encoding such a lectin (AOL) from the nematophagous fungus Arthrobotrys oligospora (deuteromycete). The deduced primary structure of the AOL gene displayed an extensive similarity (identity 46.3%) to that of a gene encoding a lectin (ABL) recently isolated from the mushroom Agaricus bisporus (basidiomycete), but not to any other fungal, microbial, plant or animal lectins. The similarities between AOL and ABL were further demonstrated by the observation that an antibody specific for AOL cross-reacted with ABL. Together with data showing that AOL has a binding specificity that is similar to that of ABL [Rosen, S., Bergström, J., Karlsson, K.-A. & Tunlid, A. (1996) Eur. J. Biochem. 238, 830-837], these results indicate that AOL and ABL are members of a novel family of saline soluble lectins present in fungi. Southern blots indicated that there is only one AOL gene in the genome encoding a subunit (monomer) of the lectin. The primary structure of AOL did not show the presence of a typical N-terminal signal sequence. Comparison of the deduced primary structure with the molecular mass of AOL as determined by electrospray mass spectrometry (16153 Da), indicated that AOL has an acetylated N-terminal but no other post-translational modifications, and that a minor isoform is formed by deamidation. Circular dichroism (CD) spectroscopy suggested that the secondary structure of AOL contains 34% beta-sheets, 21% alpha-helix, and 45% turns and coils.


Effect of screw angulation and multiple insertions on load-to-failure of polyaxial locking system.
Polyaxial locking plates rely on the alignment between the thread-to-thread connections of the screw head and the plate hole. These implants have provided substantial support for surgeons. In particular, extended screw positioning have proven to be beneficial in the fixation of challenging fractures. This study aimed to investigate the mechanical properties of ChM 5.0 ChLP polyaxial screws inserted in off-axis trajectories, including multiple insertions and to correlate these parameters with the screw head and the plate hole thread-to-thread engagement. Polyaxial locking screws were inserted into the plates at various angles (0°,10°,15°, -15° off-axis). Multiple time inserted screws were placed firstly at 15°, then 0° and finally -15° off-axis in the same plate hole. A microCT scan of the plate-hole and screw-head interface was conducted before destructive tests. Representative screws from each group were also examined by Scanning Electron Microscope. The standard insertion at 0° sustained the greatest maximum bending strength without relocation in the screw hole. Screws inserted at 10° and 15° (one time) showed a significant reduction in load-to-failure of up to 36% and 55%, (p = 0.001) (p = 0.001) respectively. Screws inserted at -15° after a maximum of three multiple insertions with angle shift, showed a total reduction in force of up to 70% (p = 0.001). A microCT analysis of thread engagement showed significant correlations. However, the results obtained for multiple insertions were highly variable. ChM 5.0 ChLP polyaxial locking system has valuable properties that foster fracture fixation, providing various surgical options. Nevertheless, the freedom of off-axis placement and multiple insertions of the screws comes at the price of reduced force. When possible surgeons should minimize the angles of insertions.


Thyroid Hormone Upregulates Hypothalamic kiss2 Gene in the Male Nile Tilapia, Oreochromis niloticus.
Kisspeptin has recently been recognized as a critical regulator of reproductive function in vertebrates. During the sexual development, kisspeptin neurons receive sex steroids feedback to trigger gonadotropin-releasing hormone (GnRH) neurons. In teleosts, a positive correlation has been found between the thyroid status and the reproductive status. However, the role of thyroid hormone in the regulation of kisspeptin system remains unknown. We cloned and characterized a gene encoding kisspeptin (kiss2) in a cichlid fish, the Nile tilapia (Oreochromis niloticus). Expression of kiss2 mRNA in the brain was analyzed by in situ hybridization. The effect of thyroid hormone (triiodothyronine, T3) and hypothyroidism with methimazole (MMI) on kiss2 and the three GnRH types (gnrh1, gnrh2, and gnrh3) mRNA expression was analyzed by real-time PCR. Expression of thyroid hormone receptor mRNAs were analyzed in laser-captured kisspeptin and GnRH neurons by RT-PCR. The kiss2 mRNA expressing cells were seen in the nucleus of the lateral recess in the hypothalamus. Intraperitoneal administration of T3 (5 μg/g body weight) to sexually mature male tilapia significantly increased kiss2 and gnrh1 mRNA levels at 24 h post injection (P < 0.001), while the treatment with an anti-thyroid, MMI (100 ppm for 6 days) significantly reduced kiss2 and gnrh1 mRNA levels (P < 0.05). gnrh2, gnrh3, and thyrotropin-releasing hormone mRNA levels were insensitive to the thyroid hormone manipulations. Furthermore, RT-PCR showed expression of thyroid hormone receptor mRNAs in laser-captured GnRH neurons but not in kiss2 neurons. This study shows that GnRH1 may be directly regulated through thyroid hormone, while the regulation of Kiss2 by T3 is more likely to be indirect.