More than 20 years' experience of left ventricular assist device implantation at a non-transplant Centre.
Over recent decades implantable left ventricular assist devices (LVAD) have increased the possibility of improved survival in patients with advanced heart failure who also benefit from a better quality of life. The aim of this retrospective survey was to review the clinical results of LVAD implantation at a low-volume non-transplant centre (Linköping, Sweden) between 1993 and 2016. Our aim was also to assess the mortality and morbidity rates associated with implantation of three LVAD versions at our centre, and to compare our results with those from transplant centres. A retrospective cohort study was performed examining the medical records of patients who had a HeartMate® (HMI, HMII, HMIII) LVAD implanted as a bridge to heart transplantation (BTT) or as destination therapy (DT) at the University Hospital, Linköping. Our main finding was a survival to heart transplantation rate of 82% among our BTT LVAD patients. The most common adverse event among our patients was infection. A higher frequency of temporary dialysis was seen in the HMII group compared to the HMI group, and the frequency of right ventricular failure was higher in our HMII material. Our data suggests that patients requiring long-term LVAD support can safely have their device implanted and cared for at a non-transplant centre.


Spatial-temporal V-Net for automatic segmentation and quantification of right ventricle on gated myocardial perfusion SPECT images.
Functional assessment of right ventricle (RV) using gated myocardial perfusion single-photon emission computed tomography (MPS) heavily relies on the precise extraction of right ventricular contours. In this paper, we present a new deep-learning-based model integrating both the spatial and temporal features in gated MPS images to perform the segmentation of the RV epicardium and endocardium. By integrating the spatial features from each cardiac frame of the gated MPS and the temporal features from the sequential cardiac frames of the gated MPS, we developed a Spatial-Temporal V-Net (ST-VNet) for automatic extraction of RV endocardial and epicardial contours. In the ST-VNet, a V-Net is employed to hierarchically extract spatial features, and convolutional long-term short-term memory (ConvLSTM) units are added to the skip-connection pathway to extract the temporal features. The input of the ST-VNet is ECG-gated sequential frames of the MPS images and the output is the probability map of the epicardial or endocardial masks. A Dice similarity coefficient (DSC) loss which penalizes the discrepancy between the model prediction and the manual annotation was adopted to optimize the segmentation model. Our segmentation model was trained and validated on a retrospective dataset with 45 subjects, and the cardiac cycle of each subject was divided into eight gates. The proposed ST-VNet achieved a DSC of 0.8914 and 0.8157 for the RV epicardium and endocardium segmentation, respectively. The mean absolute error, the mean squared error, and the Pearson correlation coefficient of the RV ejection fraction (RVEF) between the manual annotation and the model prediction were 0.0609, 0.0830, and 0.6985. Our proposed ST-VNet is an effective model for RV segmentation. It has great promise for clinical use in RV functional assessment.


[Electroencephalographic correlates of individual features of the mental efficiency of adolescents].
Evident correlations of spatial-temporal organization of the electrical brain activity with individual features of the mental working capacity were revealed in juveniles by the methods of analysis of spectra power density and of coherence function of rhythmic EEG components. EEG in juveniles with a low working mental capacity is characterized by hypersynchroneity, monofrequency, low reactivity of alpha-rhythm and high level of generalized spatial synchronization. Juveniles with high level of mental working capacity are characterized by regional specificity of the main EEG rhythm, the presence of the local dynamic foci of interconnected activity. It is suggested that the correlation of mechanisms of controlled local and non-specific generalized cortical activity is one of the main factors, determining the individual specificity of mental ability.


Mechanistic elucidation of the catalytic activity of silver nanoclusters: exploring the predominant role of electrostatic surface.
The core and the ligand shell of metal nanoclusters (MNCs) have an influential role in modulating their spectroscopic signatures and catalytic properties. The aspect of electrostatic interactions to regulate the catalytic properties of MNCs has not been comprehensively addressed to date. Our present work conclusively delineates the role of the metal core and the electrostatic surface of MNCs involved in the reduction of nitroarenes. A facile surface modification of mercaptosuccinic acid (MSA)-templated AgNCs has been selectively achieved through Mg2+ ions (Mg-AgNCs). Microscopic studies suggest that the size of Mg-AgNCs is ∼3.3 nm, which is considerably higher than that of MSA-templated AgNCs (∼1.75 nm), confirming the formation of a nano-assembled structure. Our spectroscopic and microscopic experiments revealed that the negatively charged AgNCs efficiently catalyze the reduction of 4-nitrophenol (4-NP) with a rate constant of 0.23 ± 0.01 min-1. However, upon surface modification, the catalytic efficiency almost doubles due to the formation of Mg-AgNCs. Catalysis through AgNCs and Mg-AgNCs collectively portrays the role of the core and electrostatic surfaces. Furthermore, the role of electrostatic interaction has been substantiated by varying the ionic strength of the medium, as well as employing different molecular systems. A quantitative assessment of the Debye screening length asserts the correlation between the ionic strength of the medium and the role of electrostatic interactions involved herein. This highly enhanced catalytic aspect has been utilized for the real sample analysis, wherein AgNCs unexpectedly outperform Mg-AgNCs. This approach of real sample analysis also emanates the role of electrostatics involved. This comprehensive investigation represents the influential role of the core and ligand shell of MNCs as well as the role of electrostatics on its catalytic activities, which is relevant for the rational design of highly efficient catalysts.


Outpatient rehabilitative treatment of chronic fatigue syndrome (CFS/ME).
To assess the outcome of outpatient multidisciplinary rehabilitative treatment (graded activities/exercise programme, family sessions, and supportive care) compared with supportive care alone for children and adolescents with chronic fatigue syndrome (CFS/ME). Fifty six young people (aged 9-17 years) with CFS/ME by standard criteria were followed up for 3-24 months. All subjects received supportive care. Families additionally opted to either enter the rehabilitation programme (supportive care plus graded activities/exercise programme and family sessions) or have no additional treatment. Twenty two (39%) subjects had supportive care alone and 26 (46%) entered the programme. Treatment groups were comparable at baseline in terms of age, severity and duration of illness, Wellness score, and school attendance. At end of follow up, those in the programme group had significantly higher Wellness score and school attendance than those having supportive care alone. The programme significantly reduced the overall severity of illness: after the programme, 43% had complete resolution of CFS/ME compared to only 4.5% of those having supportive care alone. The presence of depressed mood and family beliefs about the aetiology of CFS/ME were not significantly associated with outcomes. Outpatient rehabilitative treatment offers significant potential to improve the prognosis of CFS/ME in childhood and adolescence.


Searching for circles of meaning and using spiritual experiences to help parents of infants with life-threatening illness cope.
One of the major challenges parents can face is learning that their child has a life-threatening illness. A phenomenological study was completed to identify which interactions with health care providers were and were not helpful. Parents discussed what it was like to have an infant with a life-threatening illness and what helped them to cope. Parents indicated that they benefited from identifying "circles of meaning," or ways in which their infant touched another life. Parents appreciated hearing from health care providers that they felt close to their infant. This sense of connection allowed parents to reconstruct a semblance of meaning in the midst of a life-altering event. Parents were also influenced by intuitive and spiritual experiences associated with their infant's illness. Parents should be encouraged to reflect on spiritual moments and to identify how their critically ill infant influenced the lives of other family members, friends, and health care professionals.


Comparative structural analysis of the glycosylation of salivary and buccal cell proteins: innate protection against infection by Candida albicans.
Mucosal epithelial surfaces, such as line the oral cavity, are common sites of microbial colonization by bacteria, yeast and fungi. The microbial interactions involve adherence between the glycans on the host cells and the carbohydrate-binding proteins of the pathogen. Saliva constantly bathes the buccal cells of the epithelial surface of the mouth and we postulate that the sugars on the salivary glycoproteins provide an innate host immune mechanism against infection by competitively inhibiting pathogen binding to the cell membranes. The structures of the N- and O-linked oligosaccharides on the glycoproteins of saliva and buccal cell membranes were analyzed using capillary carbon liquid chromatography-electrospray ionization MS/MS. The 190 glycan structures that were characterized were qualitatively similar, but differed quantitatively, between saliva and epithelial buccal cell membrane proteins. The similar relative abundance of the terminal glycan epitope structures (e.g. ABO(H) blood group, sialylation and Lewis-type antigens) on saliva and buccal cell membrane glycoproteins indicated that the terminal N- and O-linked glycan substructures in saliva could be acting as decoy-binding receptors to competitively inhibit the attachment of pathogens to the surface of the oral mucosa. A flow cytometry-based binding assay quantified the interaction between buccal cells and the commensal oral pathogen Candida albicans. Whole saliva and released glycans from salivary proteins inhibited the interaction of C. albicans with buccal epithelial cells, confirming the protective role of the glycans on salivary glycoproteins against pathogen infection.


Time cells in the hippocampus: a new dimension for mapping memories.
Recent studies have revealed the existence of hippocampal neurons that fire at successive moments in temporally structured experiences. Several studies have shown that such temporal coding is not attributable to external events, specific behaviours or spatial dimensions of an experience. Instead, these cells represent the flow of time in specific memories and have therefore been dubbed 'time cells'. The firing properties of time cells parallel those of hippocampal place cells; time cells thus provide an additional dimension that is integrated with spatial mapping. The robust representation of both time and space in the hippocampus suggests a fundamental mechanism for organizing the elements of experience into coherent memories.


Medical management of pyometra in three red wolves (Canis rufus).
Pyometra is a serious, life-threatening disease of both domestic and non-domestic species often requiring ovariohysterectomy to preserve the life of the animal. Medical management of pyometra has been successful in domestic and non-domestic species, and the consideration of such treatment is of marked importance in a critically endangered species. Of the canids, the red wolf (Canis rufus) is second only to African hunting dogs (Lycaon pictus) in terms of the prevalence of both cystic endometrial hyperplasia and pyometra. In this report, three red wolves were medically managed for pyometra. Aside from vaginal discharge, none of the wolves exhibited clinical signs, nor were there reflective inflammatory changes in the laboratory findings. All wolves received standard treatment for pyometra, including prostaglandin F2alpha and antibiotic therapy, while one wolf was more aggressively managed with uterine lavage. Pyometra recurred in two of the treated wolves, while the most aggressively managed wolf continues to show ultrasonographic resolution 2 yr posttreatment. Aggressive medical management of pyometra should be considered a treatment option in certain red wolf females, as it may preserve the animal's reproductive potential.


H-2 restricted regulation by soluble autoantigens of the tick-borne encephalitis virus-induced autoreactive T-effector and T-suppressor lymphocytes in mice.
Soluble autoantigens (mouse red blood cells lysed by sonication) blocked in vitro the antigen-recognizing receptors of tick-borne encephalitis (TBE) virus-induced autoreactive T-lymphocytes (ARTL), effectors of the local graft-versus-host reaction (GVHR) in a syngeneic system and prevented the development of GVHR in vivo. Antigen-recognizing receptors were also found on T-suppressors (Ts) that became activated during experimental tick-borne encephalitis in mice and inhibited the activity of ARTL. The interaction between these receptors and autoantigens in vitro resulted in a loss of the ability of Ts to inhibit in vivo the ARTL-mediated GVHR. A similar result was obtained with ARTL and Ts activated in mice infected with Langat, dengue type 2 (D2) and yellow fever (strain 17D) viruses. The block of the antigen-recognizing receptors of T-cells was reversible, and not associated with lymphokine production or effector death. The block of the antigen-recognizing receptors in vitro and the loss of the corresponding T-cell function in vivo occurred provided that the donors of soluble erythrocyte antigens (SEA) and of the lymphocytes had at least one common major histocompatibility complex (MHC) haplotype. Injection SEA from donors whose H-2 complex haplotypes were identical to those of TBE-infected recipients prevented in the latter the formation of ARTL or Ts. The autoantigens inhibiting the ARTL and Ts activities seemed to be products of the MHC genes. The role of soluble H-2 autoantigens in preventing virus-induced autoimmune reaction and maintaining a state of natural immunological tolerance is discussed.


Integrated multiplexed assays of variant effect reveal cis-regulatory determinants of catechol-O-methyltransferase gene expression.
Multiplexed assays of variant effect are powerful methods to profile the consequences of rare variants on gene expression and organismal fitness. Yet, few studies have integrated several multiplexed assays to map variant effects on gene expression in coding sequences. Here, we pioneered a multiplexed assay based on polysome profiling to measure variant effects on translation at scale, uncovering single-nucleotide variants that increase and decrease ribosome load. By combining high-throughput ribosome load data with multiplexed mRNA and protein abundance readouts, we mapped the cis-regulatory landscape of thousands of catechol-O-methyltransferase (COMT) variants from RNA to protein and found numerous coding variants that alter COMT expression. Finally, we trained machine learning models to map signatures of variant effects on COMT gene expression and uncovered both directional and divergent impacts across expression layers. Our analyses reveal expression phenotypes for thousands of variants in COMT and highlight variant effects on both single and multiple layers of expression. Our findings prompt future studies that integrate several multiplexed assays for the readout of gene expression.


Effects of hypoxia and low temperature on substrate fluxes in fish: plasma metabolite concentrations are misleading.
Oxygen levels and temperature can fluctuate rapidly in aquatic environments. Ever though the effects of environmental stresses on fish metabolism have been studied extensively, information on fue kinetics is extremely limited because it relies almost exclusively on changes in substrate concentrations. The turnover rate of nonesterified fatty acids (NEFA) has never been measured in fish. Therefore, our goal was to quantify glucose and NEFA fluxes in rainbow trout acutely exposed to severe hypoxia (25% O2 saturation) or low temperature (6 degrees C for fish acclimated to 15 degrees C) by performing continuous infusions of 6-[3H]glucose and 1-[14C]palmitate in vivo. Results show that hypoxia causes a 53% decrease in NEFA turnover rate, together with a transient increase in hepatic glucose production, whereas a rapid drop in temperature induces equivalent declines in glucose, NEFA, and oxygen fluxes [temperature coefficient approximately equal to 2]. More importantly, kinetic changes in glucose and NEFA fluxes are not accompanied by interpretable changes in the plasma concentrations of these metabolites. Thus using concentration changes to draw conclusions about fluxes must be avoided.


Saccadic eye movement performance as an indicator of driving ability in elderly drivers.
Regular checking of the fitness to drive of elderly car-license holders is required in some countries, and this will become increasingly important as more countries face aging populations. The present study investigated whether the analysis of saccadic eye movements could be used as a screening method for the assessment of driving ability. Three different paradigms (prosaccades, antisaccades, and visuovisual interactive (VVI) saccades) were used to test saccadic eye movements in 144 participants split into four groups: elderly drivers who came to the attention of road authorities for suspected lack of fitness to drive, a group of elderly drivers who served as a comparison group, a group of neurology patients with established brain lesion diagnoses, and a young comparison group. The group of elderly drivers with suspected deficits in driving skills also underwent a medical examination and a practical on-road driving test. The results of the saccadic eye tests of the different groups were compared. Antisaccade results indicated a strong link to driving behaviour: elderly drivers who were not fit to drive exhibited a poor performance on the antisaccade task and the performance in the VVI task was also clearly poorer in this group. Testing saccadic eye movements appears to be a promising and efficient method for screening large numbers of people such as elderly drivers. This study indicated a link between antisaccade performance and the ability to drive. Hence, measuring saccadic eye movements should be considered as a tool for screening the fitness to drive.


Changes in soluble ICAM-1 level during hemodialysis.
Intercellular adhesion molecule-1 (ICAM-1) is expressed on the surface of various types of cells, including lymphocytes, monocytes and vascular endothelial cells. Recently, ICAM-1 was reported to be shed from the cell membranes and released into circulation. The soluble ICAM-1 (sICAM-1) level has been reported to be increased in patients with certain inflammatory diseases. It is well known that the functions of leukocytes including neutrophils, lymphocytes and monocytes are impaired in patients with chronic hemodialysis. In this study, we evaluated the effect of hemodialysis (HD) on the lymphocytes and monocytes by periodically measuring the serum concentration of sICAM-1 during HD. Pre-HD sICAM-1 levels in chronic hemodialysis patients were significantly increased as compared with healthy subjects. Two hundred and forty minutes after the start of HD, sICAM-1 levels were significantly higher than the pre-HD levels. The sICAM-1 levels at the venous side of the dialyzer were significantly increased compared with the levels at the arterial side. There was no significant difference between the sICAM-1 levels of the patients under hemodialysis with the regenerated cellulose membrane and those with the polymethylmethacrylate membrane. These results suggest that ICAM-1 is shed from the surface of mononuclear cells (lymphocytes and monocytes) and released into circulation stimulated by hemodialysis membranes. Chronic hemodialysis may impair the function of mononuclear cells by inducing the shedding of ICAM-1.


Horizontal C-1 fractures in association with unstable distraction injuries of the craniocervical junction.
Horizontal fractures of the anterior arch of the atlas not associated with odontoid fractures have been regarded as stable fractures that can be simply treated with a cervical collar. The authors report that C-1 horizontal fractures also occur in association with severe distraction injuries of the craniocervical junction. The authors retrospectively reviewed imaging studies obtained in all surviving patients in whom unstable distraction injuries of the craniocervical junction were diagnosed and in whom surgery was performed at their institution between 1995 and 2009. All patients who also had a C-1 horizontal fracture as a component of their spinal injury were included in the study. Of 47 patients with unstable craniocervical distraction injuries, 5 patients had a horizontal fracture through the anterior ring of C-1: 1 patient had a primarily C1-2 distractive injury whereas 4 others had a grossly unstable injury across the occipitoatlantal junction (occipitoatlantal dissociation). The finding of a C-1 horizontal fracture does not always reflect a benign injury and its presence should heighten the concern of a more severe and unstable injury at the craniocervical junction.


Growth of Desulfovibrio vulgaris when respiring U(VI) and characterization of biogenic uraninite.
The capacity of Desulfovibrio vulgaris to reduce U(VI) was studied previously with nongrowth conditions involving a high biomass concentration; thus, bacterial growth through respiration of U(VI) was not proven. In this study, we conducted a series of batch tests on U(VI) reduction by D. vulgaris at a low initial biomass (10 to 20 mg/L of protein) that could reveal biomass growth. D. vulgaris grew with U(VI) respiration alone, as well as with simultaneous sulfate reduction. Patterns of growth kinetics and solids production were affected by sulfate and Fe(2+). Biogenic sulfide nonenzymatically reduced 76% of the U(VI) and greatly enhanced the overall reduction rate in the absence of Fe(2+) but was rapidly scavenged by Fe(2+) to form FeS in the presence of Fe(2+). Biogenic U solids were uraninite (UO2) nanocrystallites associated with 20 mg/g biomass as protein. The crystallite thickness of UO2 was 4 to 5 nm without Fe(2+) but was <1.4 nm in the presence of Fe(2+), indicating poor crystallization inhibited by adsorbed Fe(2+) and other amorphous Fe solids, such as FeS or FeCO3. This work fills critical gaps in understanding the metabolic utilization of U by microorganisms and formation of UO2 solids in bioremediation sites.


Prognosis and Interplay of Cognitive Impairment and Sarcopenia in Older Adults Discharged from Acute Care Hospitals.
Sarcopenia and cognitive impairment are associated with an increased risk of negative outcomes, but their prognostic interplay has not been investigated so far. We aimed to investigate the prognostic interaction of sarcopenia and cognitive impairment concerning 12-month mortality among older patients discharged from acute care wards in Italy. Our series consisted of 624 patients (age = 80.1 ± 7.0 years, 56.1% women) enrolled in a prospective observational study. Sarcopenia was defined following the European Working Group on Sarcopenia in Older People (EWGSOP) criteria. Cognitive impairment was defined as age- and education-adjusted Mini-Mental State Examination (MMSE) score < 24 or recorded diagnosis of dementia. The study outcome was all-cause mortality during 12-month follow-up. The combination of sarcopenia and cognitive ability was tested against participants with intact cognitive ability and without sarcopenia. Overall, 159 patients (25.5%) were identified as having sarcopenia, and 323 (51.8%) were cognitively impaired. During the follow-up, 79 patients (12.7%) died. After adjusting for potential confounders, the combination of sarcopenia and cognitive impairment has been found associated with increased mortality (HR = 2.12, 95% CI = 1.05-4.13). Such association was also confirmed after excluding patients with dementia (HR = 2.13, 95% CI = 1.06-4.17), underweight (HR = 2.18, 95% CI = 1.03-3.91), high comorbidity burden (HR = 2.63, 95% CI = 1.09-6.32), and severe disability (HR = 2.88, 95% CI = 1.10-5.73). The co-occurrence of sarcopenia and cognitive impairment may predict 1-year mortality in older patients discharged from acute care hospitals.


Feasibility of magnetic resonance spectroscopy for evaluating fetal lung maturity.
Amniocentesis is an invasive procedure with inherent risks. Magnetic resonance (MR) spectroscopy is a safe noninvasive way of measuring levels of choline-containing compounds (including surfactant) and other metabolites. The purpose of this study was to test the feasibility of assessing fetal lung maturity in vivo and ex vivo using MR spectroscopy to determine differences in amniotic fluid choline concentrations between the second and third trimesters. Magnetic resonance spectroscopy was performed on ex vivo samples of amniotic fluid from second- and third-trimester fetuses. In vivo MR spectroscopy was performed on amniotic fluid and fetal lungs in third-trimester fetuses. Spectral acquisition and analysis were performed by an attending radiologist in conjunction with an MR spectroscopist. Choline-containing compounds were observed from 3.20 to 3.25 ppm. Comparison of spectra from second- and third-trimester amniocentesis revealed a trend toward increased choline at later gestational ages. Spectra from amniotic fluid and lungs of a third-trimester fetus showed that choline can be detected in the in vivo setting. Magnetic resonance spectroscopy is a safe noninvasive procedure that enables measurement of choline-containing compounds in fetal lung and amniotic fluid. Magnetic resonance spectroscopy shows a trend toward an increased quantity of choline in third- vs second-trimester amniocentesis.


Yeast cytochrome c with phenylalanine or tyrosine at position 87 transfers electrons to (zinc cytochrome c peroxidase)+ at a rate ten thousand times that of the serine-87 or glycine-87 variants.
Of the many factors known to influence the rate of electron transfer between two metalloproteins, it is particularly difficult to assess the role of the polypeptide matrix intervening between the donor and acceptor sites. To determine whether the phylogenetically conserved Phe-87 of yeast iso-1-cytochrome c helps to mediate electron transfer between cytochrome c and cytochrome c peroxidase, we have constructed mutants of cytochrome c that are altered at this position and now have studied the kinetics of long-range electron transfer within their complexes with zinc-substituted cytochrome c peroxidase. We find that the rate of electron transfer from reduced cytochrome c to the zinc cytochrome c peroxidase pi-cation radical is four orders of magnitude greater when phenylalanine or tyrosine is present at position 87 than when serine or glycine is present.


Fixation of the Achilles tendon insertion using suture button technology.
In the operative treatment of Achilles insertional tendinopathy, no guidelines exist concerning which form of fixation of the Achilles tendon insertion is superior. Transcalcaneal drill pin passage does not place any major plantar structures at risk, and the addition of a Krackow stitch and suture button to the fixation technique provides a significant increase in ultimate load to failure in Achilles tendon insertional repairs. Controlled laboratory study. The Achilles tendon insertions in 6 fresh-frozen cadaveric ankles were detached, and transcalcaneal drill pins were passed. Plantar dissection took place to evaluate the drill pin relationship to the plantar fascia, lateral plantar nerve and artery, flexor digitorum longus tendon, and master knot of Henry. The Achilles tendons were then repaired with a double-row suture anchor construct alone or with a suture button and Krackow stitch added to the double-row suture anchor construct. The repairs were then tested to maximum load to failure at 20 mm/min. The mode of failure was recorded, and the mean maximum load to failure was assessed using the Student t test for distributions with equal variance. Transcalcaneal drill pin passage did not place any selected anatomic structures at risk. The mean maximum load to failure for the suture bridge group was 239.2 N; it was 391.4 N for the group with the suture button (P = .014). The lateral plantar artery was the structure placed at greatest risk from drill pin placement, with a mean distance of 22.7 mm (range, 16.5-29.2 mm) between the pin and artery. In this laboratory study, transcalcaneal drill pin passage appeared to be anatomically safe, and the use of suture button technology with a Krackow stitch for Achilles tendon insertional repair significantly increased repair strength. Achilles tendon insertional repair with suture button fixation and a Krackow stitch may facilitate the earlier institution of postoperative rehabilitation and improve clinical outcomes.


Enzymatic determination of homocysteine in cell extracts.
Determination of homocysteine levels in cells and serum is important because high homocysteine is a risk factor for cardiovascular disease. The currently used methods for homocysteine analysis either are time consuming or rely on the use of expensive equipment. Described in this study is an enzymatic assay that determines levels of homocysteine in multiple samples in less than 30 min at levels from 5 to 50 pmol using only a spectrophotometer. The reproducibility of the assay is consistent with the other methods currently used. A second assay, that is about 5-fold more sensitive, follows the enzymatic catalyzed solvent exchange of protons on glycine, which requires a scintillation counter. Both the spectrophotometric and the radiometric methods are based on the conversion of 5-methyltetrahydrofolate to tetrahydrofolate by methionine synthase. The tetrahydrofolate is formed in stoichiometric amounts to the homocysteine in the sample. In the spectrophotometric method the tetrahydrofolate is used at catalytic levels by three enzymes to form a metabolic cycle that generates NADPH from NADP(+). In the radiometric assay tetrahydrofolate is required for the enzymatic exchange of the pro 2S proton of glycine with solvent. L-Cysteine, at levels more than 30-fold higher than the upper level of homocysteine used in these assays, does not give any measurable response.


Altered VEGF-stimulated Ca2+ signaling in part underlies pregnancy-adapted eNOS activity in UAEC.
In pregnancy, the uterine vasculature undergoes dramatic vasodilatory adaptations. Previously, vascular endothelial growth factor (VEGF) has been shown to stimulate endothelial nitric oxide synthase (eNOS) in uterine artery endothelial cells (UAECs) derived from pregnant ewes to a greater extent than those from non-pregnant ewes in a manner not fully explained by changes in the phosphorylation of eNOS. In this study, we used Fura-2 Ca(2+) imaging and arginine-to-citrulline conversion eNOS activity assays to assess the importance of VEGF-stimulated Ca(2+) responses in pregnancy-related changes in NO production in UAEC. In this study, we show that pregnancy-induced changes in VEGF-stimulated Ca(2+) responses could account in part for the greater capacity of VEGF to stimulate eNOS in UAECs from pregnant versus non-pregnant animals. VEGF-stimulated Ca(2+) responses in UAECs from pregnant and non-pregnant animals were mediated through VEGF receptor 2 and were detected in roughly 15% of all cells. There were no pregnancy-specific differences in area under the curve or peak height. UAECs from pregnant animals were more consistent in the time to response initiation, had a larger component of extracellular Ca(2+) entry, and were more sensitive to a submaximal dose of VEGF. In UAECs from pregnant and non-pregnant animals Ca(2+) responses and eNOS activation were sensitive to the phospholipase C/inositol 1,4,5-trisphosphate pathway inhibitors 2-aminoethoxydiphenylborane and U73122. Thus, changes in VEGF-stimulated [Ca(2+)]i are necessary for eNOS activation in UAECs, and pregnancy-induced changes in Ca(2+) responses could also in part explain the pregnancy-specific adaptive increase in eNOS activity in UAECs.


The tetrameric pheromone module SteC-MkkB-MpkB-SteD regulates asexual sporulation, sclerotia formation and aflatoxin production in Aspergillus flavus.
For eukaryotes like fungi to regulate biological responses to environmental stimuli, various signalling cascades are utilized, like the highly conserved mitogen-activated protein kinase (MAPK) pathways. In the model fungus Aspergillus nidulans, a MAPK pathway known as the pheromone module regulates development and the production of secondary metabolites (SMs). This pathway consists five proteins, the three kinases SteC, MkkB and MpkB, the adaptor SteD and the scaffold HamE. In this study, homologs of these five pheromone module proteins have been identified in the plant and human pathogenic fungus Aspergillus flavus. We have shown that a tetrameric complex consisting of the three kinases and the SteD adaptor is assembled in this species. It was observed that this complex assembles in the cytoplasm and that MpkB translocates into the nucleus. Deletion of steC, mkkB, mpkB or steD results in abolishment of both asexual sporulation and sclerotia production. This complex is required for the positive regulation of aflatoxin production and negative regulation of various SMs, including leporin B and cyclopiazonic acid (CPA), likely via MpkB interactions in the nucleus. These data highlight the conservation of the pheromone module in Aspergillus species, signifying the importance of this pathway in regulating fungal development and secondary metabolism.


Doppler fluctuation spectroscopy of intracellular dynamics in living tissue.
Intracellular dynamics in living tissue are dominated by active transport driven by bioenergetic processes far from thermal equilibrium. Intracellular constituents typically execute persistent walks. In the limit of long mean free paths, the persistent walks are ballistic, exhibiting a "Doppler edge" in light scattering fluctuation spectra. At shorter transport lengths, the fluctuations are described by lifetime-broadened Doppler spectra. Dynamic light scattering from transport in the ballistic, diffusive, or the crossover regimes is derived analytically, including the derivation of autocorrelation functions through a driven damped harmonic oscillator analog for light scattering from persistent walks. The theory is validated through Monte Carlo simulations. Experimental evidence for the Doppler edge in three-dimensional (3D) living tissue is obtained using biodynamic imaging based on low-coherence interferometry and digital holography.


Impaired mast cell-dependent natural immunity in complement C3-deficient mice.
The complement system is widely regarded as essential for normal inflammation, not least because of its ability to activate mast cells. However, recent studies have called into question the importance of complement in several examples of mast cell-dependent inflammatory responses. To investigate the role of complement in mast cell-dependent natural immunity, we examined the responses of complement-deficient mice to caecal ligation and puncture, a model of acute septic peritonitis that is dependent on mast cells and tumour necrosis factor-alpha (TNF-alpha). We found that C4- or C3-deficient mice were much more sensitive to caecal ligation and puncture than wild-type (WT) controls (100% versus 20% in 24-h mortality, respectively). C3-deficient mice also exhibited reductions in peritoneal mast cell degranulation, production of TNF-alpha, neutrophil infiltration and clearance of bacteria. Treating the C3-deficient mice with purified C3 protein enhanced activation of peritoneal mast cells, TNF-alpha production, neutrophil recruitment, opsonophagocytosis of bacteria and resistance to caecal ligation and puncture, confirming that the defects were complement-dependent. These results provide formal evidence that complement activation is essential for the full expression of innate immunity in this mast cell-dependent model of bacterial infection.


[Changes in contrast sensitivity in children with optic neuritis].
Changes of spatial contrast sensitivity (SCS) were studied in children with optic neuritis in the acute, convalescent, and remote periods of disease. A selective reduction of SCS of involved and intact eyes in the high frequency band was revealed, indicating the predominant involvement of the parvocellular subsystem of vision in optic neuritis. A correlation of SCS disorders with abnormalities of visual evoked potentials was revealed. The detected regularities indicate shifts on the part of the retrochiasmal structures in patients with optic neuritis, this being confirmed by studies making use of computer-aided tomography and magnetic imaging.


Analysis of Compensatory Movements Using a Supernumerary Robotic Hand for Upper Limb Assistance.
Recently, extratheses, aka Supernumerary Robotic Limbs (SRLs), are emerging as a new trend in the field of assistive and rehabilitation devices. We proposed the SoftHand X, a system composed of an anthropomorphic soft hand extrathesis, with a gravity support boom and a control interface for the patient. In preliminary tests, the system exhibited a positive outlook toward assisting impaired people during daily life activities and fighting learned-non-use of the impaired arm. However, similar to many robot-aided therapies, the use of the system may induce side effects that can be detrimental and worsen patients' conditions. One of the most common is the onset of alternative grasping strategies and compensatory movements, which clinicians absolutely need to counter in physical therapy. Before embarking in systematic experimentation with the SoftHand X on patients, it is essential that the system is demonstrated not to lead to an increase of compensation habits. This paper provides a detailed description of the compensatory movements performed by healthy subjects using the SoftHand X. Eleven right-handed healthy subjects were involved within an experimental protocol in which kinematic data of the upper body and EMG signals of the arm were acquired. Each subject executed tasks with and without the robotic system, considering this last situation as reference of optimal behavior. A comparison between two different configurations of the robotic hand was performed to understand if this aspect may affect the compensatory movements. Results demonstrated that the use of the apparatus reduces the range of motion of the wrist, elbow and shoulder, while it increases the range of the trunk and head movements. On the other hand, EMG analysis indicated that muscle activation was very similar among all the conditions. Results obtained suggest that the system may be used as assistive device without causing an over-use of the arm joints, and opens the way to clinical trials with patients.


Gender differences in the associations between cortisol and insulin in healthy subjects.
To investigate the role of cortisol in the etiology of insulin resistance in men and women, we examined 218 healthy non-hospitalized elderly, selected from the Rotterdam Study. Free cortisol was assessed by the ratio of fasting serum cortisol over corticosteroid-binding globulin (CBG), and insulin resistance was estimated by the fasting insulin level. CBG was higher in women and decreased with age. In both men and women, the early morning free cortisol level showed no association with age or waist/hip ratio. In men, an inverse association between cortisol and body mass index was observed. In women, higher cortisol levels were associated with increased insulin levels; an increase of 9.7 mU/l insulin per unit cortisol/CBG (S.E. 3.9, P = 0.01). The association did not change after adjustment for age, body mass index or waist/hip ratio. The results of this study in elderly subjects suggest that in women cortisol may be implicated in the age-associated insulin resistance.


KATP channels mediate adenosine-induced hyperemia in retina.
The authors and others have shown previously that the purine nucleoside adenosine is a potent vasodilator in the retinal microcirculation, mediating increases in retinal blood flow (RBF) in response to several autoregulatory stimuli. The current experiments were undertaken to elucidate the involvement of adenosine triphosphate (ATP)-sensitive potassium (KATP) channels and the adenylate cyclase--cyclic adenosine monophosphate (cAMP) second-messenger system in the transduction of adenosine's hyperemic response. Retinal fluorescein angiograms were videorecorded in isoflurane-anesthetized newborn pigs, and changes in arteriovenous transit times and retinal arteriolar and venular diameters were used to estimate stimulus-induced changes in RBF. Intravitreal perivascular microsuffusion of 5 nmol and 50 nmol adenosine caused dose-dependent increases in RBF of 79% +/- 4% (P < 0.05; n = 5) and 323% +/- 61% (P < 0.05; n = 5), respectively. The KATP channel antagonist glibenclamide (0.5 nmol and 5 nmol) caused a significant, dose-dependent attenuation of the hyperemic response to 5 nmol adenosine. The specificity of glibenclamide for blocking KATP channels was demonstrated by its ability to block by 94% +/- 6% (P < 0.05; n = 5) the increase in RBF (94% +/- 7%; P < 0.05) elicited by the intravitreal microsuffusion of the KATP channel agonist cromakalim (5 nmol), whereas it had no effect on the 103% +/- 12% increase in RBF (P < 0.05; n = 5) induced by the nitric oxide donor sodium nitroprusside (15 nmol). Adenosine-induced hyperemia was not potentiated by forskolin (1.7 nmol; n = 4), an adenylate cyclase activator, and was not attenuated by dideoxyadenosine (5 nmol; n = 4), an adenylate cyclase inhibitor. In addition, no significant increases in RBF could be elicited by 2.5 to 25 nmol 8-bromo-cAMP (n = 4), a membrane-permeable cAMP analog. These results in the piglet indicate that adenosine increases blood flow in the retina by activating KATP channels, not by increasing in cyclic AMP secondary to adenylate cyclase activation. They also underscore the potential importance of KATP channels in the transduction of retinal vasodilatative responses to other agonists.


Presence and quantification of neuropeptide Y in pulmonary edema fluids in rats.
The present study was undertaken to determine the contribution of neuropeptide Y to edema occurrence in neurogenic and hydrostatic pulmonary edema. To induce neurogenic pulmonary edema, fibrinogen and thrombin were injected into the cisterna magna; and to evoke hydrostatic pulmonary edema, saline was infused intravenously. Concentrations of neuropeptide Y in serum and edema fluid were measured using enzyme-linked immunosorbent assay (ELISA), which showed a mean value of 158 nM (95% confidence limit 124-202 nM) in the neurogenic edema fluid, significantly higher than that in the hydrostatic one. Using immunohistochemistry, fluorescent signals reactive to neuropeptide Y were found in the alveolar macrophages and edema fluid in case of fibrin-induced pulmonary edema, but were almost absent in hydrostatic edema and absent in normal lungs. Mean ratio of protein concentrations in edema fluid to that in serum was 74.9 +/- 0.9% in fibrin-induced pulmonary edema, being higher than that in hydrostatic one. From these results, we conclude that neuropeptide Y has a relationship to the high protein concentration ratio, i.e., to increased pulmonary vascular permeability, and consequently may contribute to the development of neurogenic pulmonary edema in rats.


Combinatorial design of biomaterials for drug delivery: opportunities and challenges.
The rational design of biodegradable polymeric devices for controlled drug delivery and tissue engineering is an important area of research for advancing new therapies for cancer, diabetes and immune-related disorders. In an era of escalating costs for discovery-based research, there is an urgent need to develop new and rapid methods to design drug delivery systems. By merging this field of study with rapid and high throughput methods of design, optimization and development, researchers have been able to accelerate the discovery and design processes for these devices. Combinatorial research enables the rapid identification of key regions of interest. This review focuses on the opportunities and challenges in the area of combinatorial biomaterials design for drug delivery, as there has been a great deal of significant progress over the past decade to propel this approach for the rational design of biomaterials.


What is the burden of chronic cough for families?
The burden of children's chronic cough to parents is largely unknown. The objectives of this study were as follows: (1) to determine the burden of chronic cough using a purposely designed questionnaire, and (2) to evaluate psychological (child's anxiety and parental emotional distress) and other influences on the reported burden of cough. Parents of children newly referred for chronic cough completed three questionnaires (Spence anxiety scale; depression, anxiety, and stress 21-item scale [DASS]; and burden of cough questionnaire) at enrollment. The last 79 parents also completed these questionnaires at follow-up. Median age of the 190 children recruited was 2.6 years. The number of medical consultations for coughing illness in the last 12 months was high: > 80% of children had > or = 5 doctor visits and 53% had > 10 visits. At presentation, burden scores correlated to parental DASS scores when their child was coughing. Stress was the largest contributor to parents' emotional distress. Parental anxiety and depression scores were within published norms. Scores on all three DASS subscales reduced significantly when the children ceased coughing. At follow-up, the reduction in burden scores was significantly higher in the "ceased coughing" group (n = 49) compared to the "still coughing" group (n = 32). Chronic cough in children is associated with a high burden of recurrent doctor visits, parental stress, and worries that resolve when cough ceases. Parents of children with chronic cough did not have above-average anxiety or depression levels. This study highlights the need to improve the management of children with chronic cough, including clinicians being cognizant of the emotional distress of the parents.


Potent Neutralization Ability of a Human Monoclonal Antibody Against Serotype 1 Dengue Virus.
The incidence of dengue virus (DENV) infections has been escalating in tropical and subtropical countries, but there are still no effective therapeutic options. In the present study, a DENV-1-specific human monoclonal antibody (HMAb), 1G5, isolated from single plasma cells obtained from the peripheral blood mononuclear cells of dengue patients was found to have potent neutralization activity against serotype 1 DENV (DENV-1). Its neutralization activity against DENV-2 was not as strong, and it was almost absent for DENV-3 and DENV-4. The results showed that HMAb 1G5 only binds to the envelop protein of intact DENV-1 or the envelop protein under unheated and non-reducing conditions, and that it does not bind to recombinant envelope protein. This could mean that the antibody recognizes a conformational epitope of the envelope protein. Further, the findings showed that HMAb 1G5 potently neutralizes DENV-1 in both the pre- and post-attachment phases of the virus at low concentrations. In vivo studies showed that HMAb 1G5 provides protection from DENV-1 infection in a murine model. In addition, antibody-dependent enhancement that occurs at lower doses of the antibody was completely abrogated by the introduction of Leu-to-Ala mutations (1G5-LALA) or deletion of nine amino acids (1G5-9del) in the Fc region. Therefore, HMAb 1G5 shows promise as a safe and effective agent for prophylactic and therapeutic treatment of DENV-1 infection.


Mortality and Respiratory Failure After Thoracoscopic Lung Biopsy for Interstitial Lung Disease.
Surgical lung biopsy contributes to establishing a specific diagnosis among many patients with interstitial lung disease (ILD). The risks of death and respiratory failure associated with elective thoracoscopic surgical lung biopsy, and patient characteristics associated with these outcomes, are not well understood. This is a retrospective cohort study of patients who underwent elective thoracoscopic lung biopsy for ILD between 2008 and 2014, according to The Society of Thoracic Surgeons database. The study determined the incidence of operative mortality and of postoperative respiratory failure. Multivariable models were used to identify risk factors for these adverse outcomes. Among 3,085 patients, 46 (1.5%) died before hospital discharge or within 30 days of thoracoscopic lung biopsy. Postoperative respiratory failure occurred in 90 (2.9%) patients. Significant risk factors for operative mortality among patients with ILD included a diagnosis of pulmonary hypertension, preoperative corticosteroid treatment, and low diffusion capacity. Elective thoracoscopic lung biopsy among patients with ILD is associated with a low risk of operative mortality and postoperative respiratory failure. Attention to the presence of pulmonary hypertension, preoperative corticosteroid treatment, and diffusion capacity may help inform risk stratification for thoracoscopic lung biopsy among patients with ILD.


Global hunger and climate change adaptation through international trade.
International trade enables us to exploit regional differences in climate change impacts and is increasingly regarded as a potential adaptation mechanism. Here, we focus on hunger reduction through international trade under alternative trade scenarios for a wide range of climate futures. Under the current level of trade integration, climate change would lead to up to 55 million people who are undernourished in 2050. Without adaptation through trade, the impacts of global climate change would increase to 73 million people who are undernourished (+33%). Reduction in tariffs as well as institutional and infrastructural barriers would decrease the negative impact to 20 million (-64%) people. We assess the adaptation effect of trade and climate-induced specialization patterns. The adaptation effect is strongest for hunger-affected import-dependent regions. However, in hunger-affected export-oriented regions, partial trade integration can lead to increased exports at the expense of domestic food availability. Although trade integration is a key component of adaptation, it needs sensitive implementation to benefit all regions.


Health and treatment priorities in patients with multimorbidity: report on a workshop from the European General Practice Network meeting 'Research on multimorbidity in general practice'.
Setting health and treatment priorities is necessary when caring for multiple and complex patient issues. This is already done in the doctor-patient consultation-yet implicitly rather than explicitly. The aim of this European General Practice Network workshop was to advance a consultation approach that deals with shared priority setting. The workshop was divided into three parts: (1) how to gain a comprehensive health overview for patients with multiple problems as a basis for priority setting; (2) how to establish priorities considering patient and doctor perspectives; and (3) how to practice a communication style that achieves shared priority setting. The workshop participants preferred to gain information on patients' health status using documentations from patient records rather than conducting systematic assessments. The group emphasized that medical as well as everyday life problems need to be considered when determining priorities, a procedure that requires time and resources not readily available in daily practice. Existing skills for person-centred communication with patients should be applied in order to agree on priorities. Overall it became apparent how challenging it is to arrange and prioritize an array of health problems in a consultation with patients. Existing concepts augmented by innovative systematic methods may be the way forward.


Research on multi-path dense networks for MRI spinal segmentation.
Accurate and robust segmentation of anatomical structures from magnetic resonance images is valuable in many computer-aided clinical tasks. Traditional codec networks are not satisfactory because of their low accuracy of edge segmentation, the low recognition rate of the target, and loss of detailed information. To address these problems, this study proposes a series of improved models for semantic segmentation and progressively optimizes them from the three aspects of convolution module, codec unit, and feature fusion. Instead of the standard convolution structure, we apply a new type of convolution module for the feature extraction. The networks integrate a multi-path method to obtain richer-detail edge information. Finally, a dense network is utilized to strengthen the ability of the feature fusion and integrate more different-level information. The evaluation of the Accuracy, Dice coefficient, and Jaccard index led to values of 0.9855, 0.9185, and 0.8507, respectively. These metrics of the best network increased by 1.0%, 4.0%, and 6.1%, respectively. Boundary F1-Score reached 0.9124 indicating that the proposed networks can segment smaller targets to obtain smoother edges. Our methods obtain more key information than traditional methods and achieve superiority in segmentation performance.


Photocatalytic conversion of CO2 over graphene-based composites: current status and future perspective.
The continuous rise in the atmospheric CO2 level and the ever-increasing demand of energy have raised serious concerns about the ensuing effects on the global climate change and future energy supply. Photocatalytic conversion of CO2, which uses solar light energy to recycle CO2 into fuels and chemicals, provides a promising and straightforward strategy to simultaneously reduce the atmospheric CO2 level and fulfil the future energy demand. However, the lack of substantial development of state-of-the-art materials remains a major bottleneck of this technology. In recent years, graphene-based composite photocatalysts have gained increasing interest in CO2 conversion due to the introduction of graphene with a series of unique physicochemical properties, which has shown to play a significant role in promoting the photocatalytic solar energy conversion efficiency. In this review, we comprehensively summarize the typical literature reports on graphene-based composites for photocatalytic conversion of CO2 to produce solar fuels and chemicals. The main types of the reported graphene-based composites and the role of graphene in the composites in improving the photocatalytic performance have been elaborated. In particular, we have highlighted the possible role of graphene in tuning the product selectivity of photocatalytic reduction of CO2. Finally, perspectives on the existing problems and future research on graphene-based composites toward photocatalytic conversion of CO2 are critically discussed.


Bacterial Nitric Oxide Synthase Is Required for the Staphylococcus aureus Response to Heme Stress.
Staphylococcus aureus is a pathogen that causes significant morbidity and mortality worldwide. Within the vertebrate host, S. aureus requires heme as a nutrient iron source and as a cofactor for multiple cellular processes. Although required for pathogenesis, excess heme is toxic. S. aureus employs a two-component system, the heme sensor system (HssRS), to sense and protect against heme toxicity. Upon activation, HssRS induces the expression of the heme-regulated transporter (HrtAB), an efflux pump that alleviates heme toxicity. The ability to sense and respond to heme is critical for the pathogenesis of numerous Gram-positive organisms, yet the mechanism of heme sensing remains unknown. Compound '3981 was identified in a high-throughput screen as an activator of staphylococcal HssRS that triggers HssRS independently of heme accumulation. '3981 is toxic to S. aureus; however, derivatives of '3981 were synthesized that lack toxicity while retaining HssRS activation, enabling the interrogation of the heme stress response without confounding toxic effects of the parent molecule. Using '3981 derivatives as probes of the heme stress response, numerous genes required for '3981-induced activation of HssRS were uncovered. Specifically, multiple genes involved in the production of nitric oxide were identified, including the gene encoding bacterial nitric oxide synthase (bNOS). bNOS protects S. aureus from oxidative stress imposed by heme. Taken together, this work identifies bNOS as crucial for the S. aureus heme stress response, providing evidence that nitric oxide synthesis and heme sensing are intertwined.


Multi-step down-regulation of the secretory pathway in mitosis: a fresh perspective on protein trafficking.
The secretory pathway delivers proteins synthesized at the rough endoplasmic reticulum (RER) to various subcellular locations via the Golgi apparatus. Currently, efforts are focused on understanding the molecular machineries driving individual processes at the RER and Golgi that package, modify and transport proteins. However, studies are routinely performed using non-dividing cells. This obscures the critical issue of how the secretory pathway is affected by cell division. Indeed, several studies have indicated that protein trafficking is down-regulated during mitosis. Moreover, the RER and Golgi apparatus exhibit gross reorganization in mitosis. Here I provide a relatively neglected perspective of how the mitotic cyclin-dependent kinase (CDK1) could regulate various stages of the secretory pathway. I highlight several aspects of the mitotic control of protein trafficking that remain unresolved and suggest that further studies on how the mitotic CDK1 influences the secretory pathway are necessary to obtain a deeper understanding of protein transport.


Conservative and dietary interventions for cancer-related lymphedema: a systematic review and meta-analysis.
The findings support the use of compression garments and compression bandaging for reducing lymphedema volume in upper and lower extremity cancer-related lymphedema. Specific to breast cancer, a statistically significant, clinically small beneficial effect was found from the addition of manual lymph drainage massage to compression therapy for upper extremity lymphedema volume.


Biochemistry and molecular biology of exocellular fungal beta-(1,3)- and beta-(1,6)-glucanases.
Many fungi produce exocellular beta-glucan-degrading enzymes, the beta-glucanases including the noncellulolytic beta-(1,3)- and beta-(1,6)-glucanases, degrading beta-(1,3)- and beta-(1,6)-glucans. An ability to purify several exocellular beta-glucanases attacking the same linkage type from a single fungus is common, although unlike the beta-1,3-glucanases, production of multiple beta-1,6-glucanases is quite rare in fungi. Reasons for this multiplicity remain unclear and the multiple forms may not be genetically different but arise by posttranslational glycosylation or proteolytic degradation of the single enzyme. How their synthesis is regulated, and whether each form is regulated differentially also needs clarifying. Their industrial potential will only be realized when the genes encoding them are cloned and expressed in large quantities. This review considers what is known in molecular terms about their multiplicity of occurrence, regulation of synthesis and phylogenetic diversity. It discusses how this information assists in understanding their functions in the fungi producing them. It deals largely with exocellular beta-glucanases which here refers to those recoverable after the cells are removed, since those associated with fungal cell walls have been reviewed recently by Adams (2004). It also updates the earlier review by Pitson et al. (1993).


Building branched tissue structures: from single cell guidance to coordinated construction.
Branched networks are ubiquitous throughout nature, particularly found in tissues that require large surface area within a restricted volume. Many tissues with a branched architecture, such as the vasculature, kidney, mammary gland, lung and nervous system, function to exchange fluids, gases and information throughout the body of an organism. The generation of branched tissues requires regulation of branch site specification, initiation and elongation. Branching events often require the coordination of many cells to build a tissue network for material exchange. Recent evidence has emerged suggesting that cell cooperativity scales with the number of cells actively contributing to branching events. Here, we compare mechanisms that regulate branching, focusing on how cell cohorts behave in a coordinated manner to build branched tissues.This article is part of the themed issue 'Systems morphodynamics: understanding the development of tissue hardware'.


Synergistic formation and stabilization of oil-in-water emulsions by a weakly interacting mixture of zwitterionic surfactant and silica nanoparticles.
Oil-in-water emulsions were formed and stabilized at low amphiphile concentrations by combining hydrophilic nanoparticles (NPs) (i.e., bare colloidal silica) with a weakly interacting zwitterionic surfactant, caprylamidopropyl betaine, to generate a high hydrophilic-lipophilic balance. The weak interaction of the NPs with surfactant was quantified with contact angle measurements. Emulsions were characterized by static light scattering to determine the droplet size distributions, optical photography to quantify phase separation due to creaming, and both optical and electron microscopy to determine emulsion microstructure. The NPs and surfactant acted synergistically to produce finer emulsions with a greater stability to coalescence relative to the behavior with either NPs or surfactant alone. As a consequence of the weak adsorption of the highly hydrophilic surfactant on the anionic NPs along with the high critical micelle concentration, an unusually large surfactant concentration was available to adsorb at the oil-water interface and lower the interfacial tension. The synergy for emulsion formation and stabilization for the two amphiphiles was even greater in the case of a high-salinity synthetic seawater aqueous phase. Here, higher NP adsorption at the oil-water interface was caused by electrostatic screening of interactions between (1) NPs and the anionic oil-water interface and (2) between the NPs. This greater adsorption as well as partial flocculation of the NPs provided a more efficient barrier to droplet coalescence.


Prevention of prostate cancer with finasteride: US/European perspective.
Prostate cancer is the most common male cancer, affecting one man in six. Prevention of this disease, even in a subset of patients would have a significant impact on public health. The results of the National Cancer Institute-sponsored Prostate Cancer Prevention Trial demonstrate that finasteride causes a substantial risk reduction across all known risk groups. We herein amplify on the results of this trial to assist patients and physicians in reaching individualized decisions. The results of the Prostate Cancer Prevention Trial were reviewed. The PCPT demonstrated a 24.8% reduction in prostate cancer risk with the administration of finasteride. High-grade cancers were noted in 6.4% of finasteride patients compared to 5.1% of men receiving placebo. The increase in high-grade tumors was seen within one year of finasteride exposure and did not increase over time. This observation, combined with previous evidence demonstrating an alteration in cytologic and architectural features of prostate cancer with hormonal therapy suggests that the differences in high-grade disease may be an artifact and that application of Gleason grading to these tumors may not be appropriate. Men should be presented the benefits and risks of taking finasteride and be assisted in integrating their sexual and urinary symptoms into their decision-making process. Blanket statements for or against the use of this medication ignore patient preferences and differential risk-benefit profiles from finasteride.


Immunization with host-type CD8{alpha}+ dendritic cells reduces experimental acute GVHD in an IL-10-dependent manner.
Little is known about the role of active immunization in suppressing undesirable immune responses. Because CD8alpha(+) dendritic cells (DCs) suppress certain immune responses, we tested the hypothesis that immunization of donors with host-derived CD8alpha(+) DCs will reduce host-specific donor T-cell responses. BALB/c T cells from the animals that were immunized with B6 CD8alpha(+) DCs demonstrated, in vitro and in vivo, significantly reduced proliferation and secretion of inflammatory cytokines but showed enhanced secretion of interleukin-10 (IL-10). The responses against third-party and model antigens were preserved demonstrating antigen specificity. The in vivo relevance was further demonstrated by the reduction on graft-versus-host disease (GVHD) in both a major histocompatibility complex-mismatched clinically relevant BALB/c --> B6 model and major histocompatibility complex-matched, minor-mismatched C3H.SW --> B6 model of GVHD. Immunization of the donors that were deficient in IL-10 (IL-10(-/-)) or with CD8alpha(+) DCs from B6 class II (class II(-/-)) failed to reduce T-cell responses, demonstrating (1) a critical role for secretion of IL-10 by donor T cells and (2) a direct contact between the T cells and the CD8alpha(+) DCs. Together, these data may represent a novel strategy for reducing GVHD and suggest a broad counterintuitive role for vaccination strategies in mitigating undesirable immune responses in an antigen-specific manner.


Rapid in vitro prototyping of O-methyltransferases for pathway applications in Escherichia coli.
O-Methyltransferases are ubiquitous enzymes involved in biosynthetic pathways for secondary metabolites such as bacterial antibiotics, human catecholamine neurotransmitters, and plant phenylpropanoids. While thousands of putative O-methyltransferases are found in sequence databases, few examples are functionally characterized. From a pathway engineering perspective, however, it is crucial to know the substrate and product ranges of the respective enzymes to fully exploit their catalytic power. In this study, we developed an in vitro prototyping workflow that allowed us to screen ∼30 enzymes against five substrates in 3 days with high reproducibility. We combined in vitro transcription/translation of the genes of interest with a microliter-scale enzymatic assay in 96-well plates. The substrate conversion was indirectly measured by quantifying the consumption of the S-adenosyl-L-methionine co-factor by time-resolved fluorescence resonance energy transfer rather than time-consuming product analysis by chromatography. This workflow allowed us to rapidly prototype thus far uncharacterized O-methyltransferases for future use as biocatalysts.


Two-dimensional spin-exchange solid-state NMR studies of 13 C-enriched wood.
Two-dimensional high-resolution solid-state NMR has been used to study 13C-enriched wood. Wood is a complex material containing three major polymers: cellulose, hemicelluloses and lignin. The use of an enriched 13C-compound allows the observation of intra-molecular spin-diffusion driven by dipolar couplings. Correlations between spins at progressively longer distances have been obtained as the mixing time is increased, corresponding, for each of the separate polymer chains, to intra-unit and then to inter-unit interactions, and in the case of cellulose to inter-chain interactions. A straightforward qualitative analysis of the spin diffusion spectra is shown to yield the assignment of the carbon-13 spectrum. The cellulose resonances can all be sequentially assigned using the spin diffusion experiment. Using the experiments it is shown that, at least on a distance scale of several nanometres explored by the spin diffusion process, the three main components of wood occur in separate phases. Also, a question concerning the structure of the hemicellulose units is resolved by locating the O-acetyl group at the 2-position of the xylan chains.


Evaluation and use of three cowside tests for detection of subclinical ketosis in early postpartum cows.
The objective was to evaluate the performance of 3 cowside diagnostic tests for detection of subclinical ketosis, defined as a serum beta-hydroxybutyrate (BHBA) concentration >or=1400 micromol/L. On 16 d over a 5-mo period, samples of serum, milk, and urine were collected on a large dairy facility from cows of all parities between 2 and 15 DIM. The sample proportion of subclinical ketosis was 7.6% (n = 859 samples from 545 cows). The KetoCheck powder (Great States Animal Health, St. Joseph, MO) detecting acetoacetate in milk samples was very specific (99%) but poorly sensitive (41%). Respective sensitivities and specificities of the Ketostix strip detecting acetoacetate in urine samples (Bayer Corporation, Elkhart, IN) were 78 and 96% with a cut-off point of "small", or 49 and 99% with a cut-off of "moderate." The KetoTest strip (Sanwa Kagaku Kenkyusho Co. Ltd., Nagoya, Japan) using milk samples had a sensitivity and specificity of 73 and 96% with a cut-off of 100 micromol of BHBA/L or 27 and 99% with a cut-off of 200 micromol of BHBA/L. On average, use of the Ketostix at the "small" cut-off point or the KetoTest at 100 micromol/L would result in no more than 3 or 4 false positives per 100 cows screened, with prevalence levels ranging from 5 to 30%, whereas the number of false negatives would range from one false negative at 5% prevalence to 7 or 8 false negatives at 30% prevalence. Either the Ketostix or KetoTest strips would provide acceptable results for screening individual cows on commercial dairies to detect subclinical ketosis. Over this prevalence range, the KetoCheck powder test would have limited application as a screening test. Despite only one false positive per 100 animals screened, false negatives resulting from screening with the KetoCheck test would be too frequent, ranging from 3 false negatives at 5% prevalence to 18 at 30% prevalence in a population of 100 tested cows. Finally, given their relative imprecision, use of any of these individual cowside tests to estimate herd prevalence must be done cautiously, especially when only a small number of animals are sampled.


Strain and process development for poly(3HB-co-3HP) fermentation by engineered Shimwellia blattae from glycerol.
Poly(3-hydroxybytyrate-co-3-hydroxypropionate), poly(3HB-co-3HP), is a possible alternative to synthetic polymers such as polypropylene, polystyrene and polyethylene due to its low crystallinity and fragility. We already reported that recombinant strains of Shimwellia blattae expressing 1,3-propanediol dehydrogenase DhaT as well as aldehyde dehydrogenase AldD of Pseudomonas putida KT2442, propionate-CoA transferase Pct of Clostridium propionicum X2 and PHA synthase PhaC1 of Ralstonia eutropha H16 are able to accumulate up to 14.5% (wtPHA/wtCDW) of poly(3-hydroxypropionate), poly(3HP), homopolymer from glycerol as a sole carbon source (Appl Microbiol Biotechnol 98:7409-7422, 2014a). However, the cell density was rather low. In this study, we optimized the medium aiming at a more efficient PHA synthesis, and we engineered a S. blattae strain accumulating poly(3HB-co-3HP) with varying contents of the constituent 3-hydroxypropionate (3HP) depending on the cultivation conditions. Consequently, 7.12, 0.77 and 0.32 gPHA/L of poly(3HB-co-3HP) containing 2.1, 8.3 and 18.1 mol% 3HP under anaerobic/aerobic (the first 24 hours under anaerobic condition, thereafter, aerobic condition), low aeration/agitation (the minimum stirring rate required in medium mixing and small amount of aeration) and anaerobic conditions (the minimum stirring rate required in medium mixing without aeration), respectively, were synthesized from glycerol by the genetically modified S. blattae ATCC33430 strains in optimized culture medium.


The management of gastric polyps.
Gastric polyps are important as some have malignant potential. If such polyps are left untreated, gastric cancer may result. The malignant potential depends on the histological type of the polyp. The literature base is relatively weak and any recommendations made must be viewed in light of this. Gastric polyps are sessile or pedunculated lesions that originate in the gastric epithelium or submucosa and protrude into the stomach lumen. Depending on histological type, some gastric polyps (adenomas and hyperplastic polyps) have malignant potential and are precursors of early gastric cancer. They may also indicate an increased risk of intestinal or extra-intestinal malignancy.


Ankle-arm index as a predictor of cardiovascular disease and mortality in the Cardiovascular Health Study. The Cardiovascular Health Study Group.
Peripheral arterial disease (PAD) in the legs, measured noninvasively by the ankle-arm index (AAI) is associated with clinically manifest cardiovascular disease (CVD) and its risk factors. To determine risk of total mortality, coronary heart disease, or stroke mortality and incident versus recurrent CVD associated with a low AAI, we examined the relationship of the AAI to subsequent CVD events in 5888 older adults with and without CVD. The AAI was measured in 5888 participants >/=65 years old at the baseline examination of the Cardiovascular Health Study. All participants had a detailed assessment of prevalent CVD and were contacted every 6 months for total mortality and CVD events (including CVD mortality, fatal and nonfatal myocardial infarction, congestive heart failure, angina, stroke, and hospitalized PAD). The crude mortality rate at 6 years was highest (32.3%) in those participants with prevalent CVD and a low AAI (P<0.9), and it was lowest in those with neither of these findings (8.7%, P<0.01). Similar patterns emerged from analysis of recurrent CVD and incident CVD. The risk for incident congestive heart failure (relative risk [RR]=1.61) and for total mortality (RR=1.62) in those without CVD at baseline but with a low AAI remained significantly elevated after adjustment for cardiovascular risk factors. Hospitalized PAD events occurred months to years after the AAI was measured, with an adjusted RR of 5.55 (95% CI, 3.08 to 9.98) in those at risk for incident events. A statistically significant decline in survival was seen at each 0.1 decrement in the AAI. An AAI of <0.9 is an independent risk factor for incident CVD, recurrent CVD, and mortality in this group of older adults in the Cardiovascular Health Study.


Across ages and places: Unpredictability of maternal sensory signals and child internalizing behaviors.
Patterns of sensory inputs early in life play an integral role in shaping the maturation of neural circuits, including those implicated in emotion and cognition. In both experimental animal models and observational human research, unpredictable sensory signals have been linked to aberrant developmental outcomes, including poor memory and effortful control. These findings suggest that sensitivity to unpredictable sensory signals is conserved across species and sculpts the developing brain. The current study provides a novel investigation of unpredictable maternal sensory signals in early life and child internalizing behaviors. We tested these associations in three independent cohorts to probe the generalizability of associations across continents and cultures. The three prospective longitudinal cohorts were based in Orange, USA (n = 163, 47.2 % female, Mage = 1 year); Turku, Finland (n = 239, 44.8 % female, Mage = 5 years); and Irvine, USA (n = 129, 43.4 % female, Mage = 9.6 years). Unpredictability of maternal sensory signals was quantified during free-play interactions. Child internalizing behaviors were measured via parent report (Orange & Turku) and child self-report (Irvine). Early life exposure to unpredictable maternal sensory signals was associated with greater child fearfulness/anxiety in all three cohorts, above and beyond maternal sensitivity and sociodemographic factors. The association between unpredictable maternal sensory signals and child sadness/depression was relatively weaker and did not reach traditional thresholds for statistical significance. The correlational design limits our ability to make causal inferences. Findings across the three diverse cohorts suggest that unpredictable maternal signals early in life shape the development of internalizing behaviors, particularly fearfulness and anxiety.


Massive blood transfusion.
Pediatricians in the hospital setting must frequently treat children who require massive transfusion (MT) in a variety of clinical situations ranging from major trauma to neonatal hyperbilirubinemia. After identifying the need for massive transfusion, the pediatrician must select the appropriate blood components. Different blood components have specific temperature, preservative, and time requirements for their storage. Changes, termed storage lesions, occur over time in blood components during storage; biochemical changes include decreased levels of 2,3-DPG, a decrease in pH, and an increase in supernatant potassium (K+) with a concurrent decrease in intracellular K+. These changes may affect the function and the viability of components. Additionally, physical changes such as deformation of the red cell membrane occur during storage. Knowledge of these storage lesions is necessary for the pediatrician to make the most appropriate decisions regarding the preparation and selection of components during MT. Serious complications of MT include hemostatic abnormalities, biochemical/metabolic abnormalities, hypothermia, mechanical injury and the effect of Rh incompatibility, each of which has a specific management response. Pediatricians need to be aware of the potential complications associated with massive transfusion, to take measures to prevent them when possible, to anticipate additional transfusion requirements, and to know how to manage them in the pediatric patient.


Psychophysiological and endocrine responses to sexual arousal in women.
The unexplored possibility that a sexually induced endocrine response might prime further sexual arousal in women guided the current investigation. Healthy, premenopausal, heterosexual women in the follicular phase of their menstrual cycle were randomly assigned to either an experimental or a control group. The experimental group was exposed to a sexually explicit videotape, while the control group saw a nonerotic videotape. Ninety minutes later both groups saw a sexually explicit videotape. Vaginal vasocongestion and hormones (cortisol, prolactin, luteinizing hormone, testosterone) were measured continuously and subjective responses were sampled at 20-min intervals. Compared to controls, experimental subjects showed a greater amplitude and longer duration vaginal response to the second videotape. Subjective measures showed greater sexual response to the second erotic videotape compared to the first, an effect that was not mediated by the hormones measured here. Prolactin decreased significantly across the session for both groups, and several behavioral and affective responses were significantly correlated with hormonal levels. Commonalities and divergence with results of prior research point to the complexity and subtlety of endocrine interactions with sexual response as well as likely sex differences in hormone-behavioral interactions.


Measuring quality of care in patients with nonvariceal upper gastrointestinal hemorrhage: development of an explicit quality indicator set.
With an increasing emphasis on quality in health care and recognition of inconsistencies in the management of patients with nonvariceal upper gastrointestinal hemorrhage (NVUGIH), it is critical to establish a set of explicit quality indicators (QIs) in NVUGIH. We conducted a nine-member, multidisciplinary expert panel and followed modified Delphi methods to systematically identify a set of QIs for NVUGIH. The panel performed independent ratings of each candidate QI using a nine-point RAND appropriateness scale, then met in person and re-voted using an iterative process of discussion. The final set comprised QIs with a median RAND Appropriateness Score >or=7 and no disagreement among experts. Among 116 candidate QIs, the panel rated 26 as valid measures of quality care. The selected QIs cover pre-endoscopy, endoscopy, and post-endoscopy care, including diagnosis, early resuscitation, risk stratification, endoscopic care, Helicobacter pylori management, and proton pump inhibitor therapy. We have developed an explicit set of evidence-based QIs in NVUGIH, providing physicians and institutions with a tool to identify processes amenable to quality improvement. This tool is intended to be applicable in all institutions providing care for NVUGIH patients.


Hypersensitivity pneumonitis and alpha-chemokines.
Hypersensitivity pneumonitis (HP) is categorized as a Type-1 helper (Th1) disease. The resulting granuloma formation is dependent on T cells and the Th1 cytokine interferon (IFN)-γ. In experimental setting, the production of IFN-γ-induced protein 10 (IP-10), monokine induced by IFN-γ (MIG), IFN-inducible T-cell-alpha chemoattractant (I-TAC), has been shown (in mice exposed to the particulate antigens that cause HP) during the development of HP. The production of these chemokines was associated with an influx of chemokine (C-X-C motif) receptor (CXCR)3 CXCR3(+)/CD4(+) T cells into lungs. This suggests that IFN-γ mediates the recruitment of CXCR3(+)/CD4(+) T cells into the lung via the production of IP-10, MIG, and I-TAC, resulting in granuloma formation. In humans it has been shown that lymphocytes infiltrating lung biopsies are CD8 T cells for CXCR3. Furthermore, the T cells accumulating in the bronchoalveolar lavage (BAL) of HP were CXCR3(+)/IFN-γ(+) type 1 CD8(+) T cells (Tc1) exhibiting a strong in vitro migratory capability in response to IP-10. Alveolar macrophages express and secrete, in response to IFN-γ, high levels of IP-10, capable of inducing chemotaxis of the CXCR3(+) T-cell line. High levels of CXCR3 ligands were shown in the fluid of the BAL in HP patients. These data confirm that IFN-γ mediates the recruitment of lymphocytes into the lung via production of IP-10, resulting in Tc1-cell alveolitis and granuloma formation. It has been suggested that differences in the expression of CXC chemokines and Th1 cytokines may contribute to different immunopathogenesis, clinical course and responsiveness to treatment of HP.


A Network Comparative Meta-analysis of Percutaneous Dilatational Tracheostomies Using Anatomic Landmarks, Bronchoscopic, and Ultrasound Guidance Versus Open Surgical Tracheostomy.
Several different tracheostomy techniques (percutaneous and surgical) have been studied extensively in previous direct pairwise meta-analyses. However, a network comparative meta-analysis comparing all has not been conducted before. We sought to compare three percutaneous dilatational tracheostomy techniques with open surgical tracheostomy technique (performed in the operating room or in the intensive care unit by bedside) in terms of their association with procedure-related major complications and procedure time. We searched PubMed and Cochrane register of randomized active comparator trials. A network comparative meta-analysis was performed in Stata using frequentist methodology. Major complications were defined as a composite of a priori-selected procedure-related complications. Tracheostomy techniques that did not require any direct bronchoscopic or ultrasonographic visualization of the entire procedure were grouped under the heading-anatomic landmark-based dilatational tracheostomy (ALDT). This along with bronchoscopic-guided dilatational tracheostomy (BDT), ultrasound-guided (UDT), and surgical tracheostomy (SGT) were compared with each other using network meta-analysis in Stata after all major assumptions (similarity, transitivity, and consistency) for performing a network were met. Log odds ratio (and standard errors) of the comparison of major complications between any two tracheostomy techniques (using indirect estimates) was statistically insignificant. Pairwise meta-analysis showed significant differences in procedure times between SGT and ALDT [mean difference: 9.96 min (SE 3.18)] and between SGT and BDT [15.67 min (SE 3.85)]. The indirect network meta-analysis comparing one versus the other also showed a statistically significant time difference between surgical tracheostomy when compared with every other technique. The results of our network meta-analysis show that all tracheostomy techniques are comparable with respect to associated procedure-related complications, but all three percutaneous techniques take far less procedure time compared to the surgical tracheostomy.


Unilateral and bilateral ECT: a study of memory disturbance and relief from depression.
Fifty-one endogenous and reactive female depressives were given a course of either unilateral non-dominant, unilateral dominant, or bilateral ECT. Visual and verbal memory tests and confusion ratings were administered at frequent intervals during the treatment course. Pre-and post-treatment assessments of depression were made. Comparisons of the therapeutic effect of six and of eight ECTs were studied separately. One month after the last ECT the patients were again assessed on the memory and the depression tests. The results of the memory tests indicate that unilateral non-dominant ECT produced least memory disturbance (particularly of a verbal kind) and also less immediate confusion within 40 minutes of each ECT. This observation applies more to the reactive than the endogenous group. Comparisons of the depression tests reveal that unilateral non-dominant ECT is as effective in relieving depression as bilateral ECT, though progress may be less rapid. The observation holds true only for the reactive depressives. Endogenous depressives benefit more from bilateral ECT. Caution is advised against the administration of unilateral dominant ECT, since this group does not respond to treatment as well as the other two groups. Degree of improvement as a whole does not appear to be related to the degree of confusion experienced. The implications of these findings are discussed.


Evaluation of a hydrogel membrane on bone regeneration in furcation periodontal defects in dogs.
The aim of the study was to evaluate bone regeneration using a canine model with surgically created periodontal defects filled for 12 weeks using a stratified biomaterial consisting in a biphasic calcium phosphate (BCP) covered with a crosslinking hydrogel acting as polymer membrane of silated hydroxypropyl methylcellulose (Si-HPMC) as the tested new concept. Bilateral, critical-sized, defects were surgically created at the mandibular premolar teeth of six adult beagle dogs. The defects were randomly allocated and: (i) left empty for spontaneous healing or filled with: (ii) BCP and a collagen membrane; (iii) BCP and hydrogel Si-HPMC membrane. At 12 weeks, the experimental conditions resulted in significantly enhanced bone regeneration in the test BCP/Si-HPMC group. Within the limits of this study, we suggest that the hydrogel Si-HPMC may act as an occlusive barrier to protect bone area from soft connective tissue invasion and then effectively contribute to enhance bone regeneration.


The behavioural response of slugs and snails to novel molluscicides, irritants and repellents.
The behavioural response of the slug Deroceras panormitanum (Lessona and Pollonera) and the snail Oxyloma pfeifferi (Rossmässler) to novel molluscicides was investigated in choice and no-choice experiments. Low-light video-recording in combination with automated tracking and event recording was used to identify the repellent and irritant effects of (1) cinnamamide, (2) copper ammonium carbonate, (3) a mulch, (4) a horticultural ground-cover matting impregnated with a copper formulation and (5) urea/formaldehyde. In the no-choice experiments the products had a stronger irritant effect on the snails than on the slugs. All products tested except the mulch significantly reduced the locomotor activity of both the slugs and snails. The most effective product, cinnamamide, reduced snail locomotor activity by 94% and track length by 96%. The overall repellent effect of the treatments in the choice experiments was stronger in the slugs; where presence, locomotor activity and track length in the treated area were significantly reduced by all products. The avoidance of treated areas exceeded 95% with the mulch (for slugs) and with copper ammonium carbonate (for snails).


Calvarial osteoblast gene expression in patients with craniosynostosis leads to novel polygenic mouse model.
Craniosynostosis is the premature fusion of the sutures of the calvaria and is principally designated as being either syndromic (demonstrating characteristic extracranial malformations) or non-syndromic. While many forms of syndromic craniosynostosis are known to be caused by specific mutations, the genetic etiology of non-syndromic, single-suture craniosynostosis (SSC) is poorly understood. Based on the low recurrence rate (4-7%) and the fact that recurrent mutations have not been identified for most cases of SSC, we propose that some cases of isolated, single suture craniosynostosis may be polygenic. Previous work in our lab identified a disproportionately high number of rare and novel gain-of-function IGF1R variants in patients with SSC as compared to controls. Building upon this result, we used expression array data from calvarial osteoblasts isolated from infants with and without SSC to ascertain correlations between high IGF1 expression and expression of other osteogenic genes of interest. We identified a positive correlation between increased expression of IGF1 and RUNX2, a gene known to cause SSC with increased gene dosage. Subsequent phosphorylation assays revealed that osteoblast cell lines from cases with high IGF1 expression demonstrated inhibition of GSK3β, a serine/threonine kinase known to inhibit RUNX2, thus activating osteogenesis through the IRS1-mediated Akt pathway. With these findings, we have utilized established mouse strains to examine a novel model of polygenic inheritance (a phenotype influenced by more than one gene) of SSC. Compound heterozygous mice with selective disinhibition of RUNX2 and either overexpression of IGF1 or loss of function of GSK3β demonstrated an increase in the frequency and severity of synostosis as compared to mice with the RUNX2 disinhibition alone. These polygenic mouse models reinforce, in-vivo, that the combination of activation of the IGF1 pathway and disinhibition of the RUNX2 pathway leads to an increased risk of developing craniosynostosis and serves as a model of human SSC.


Redundant roles of Sox17 and Sox18 in postnatal angiogenesis in mice.
Sox7, Sox17 and Sox18 constitute group F of the Sox family of HMG box transcription factor genes. Dominant-negative mutations in Sox18 underlie the cardiovascular defects observed in ragged mutant mice. By contrast, Sox18(-/-) mice are viable and fertile, and display no appreciable anomaly in their vasculature, suggesting functional compensation by the two other SoxF genes. Here, we provide direct evidence for redundant function of Sox17 and Sox18 in postnatal neovascularization by generating Sox17(+/-) -Sox18(-/-) double mutant mice. Whereas Sox18(-/-) and Sox17(+/-) -Sox18(+/-) mice showed no vascular defects, approximately half of the Sox17(+/-) -Sox18(-/-) pups died before postnatal day 21 (P21). They showed reduced neovascularization in the liver sinusoids and kidney outer medulla vasa recta at P7, which most likely caused the ischemic necrosis observed by P14 in hepatocytes and renal tubular epithelia. Those that survived to adulthood showed similar, but milder, vascular anomalies in both liver and kidney, and females were infertile with varying degrees of vascular abnormalities in the reproductive organs. These anomalies corresponded with sites of expression of Sox7 and Sox17 in the developing postnatal vasculature. In vitro angiogenesis assays, using primary endothelial cells isolated from the P7 livers, showed that the Sox17(+/-) -Sox18(-/-) endothelial cells were defective in endothelial sprouting and remodeling of the vasculature in a phenotype-dependent manner. Therefore, our findings indicate that Sox17 and Sox18, and possibly all three SoxF genes, are cooperatively involved in mammalian vascular development.


Dietary intake of key nutrients and subarachnoid hemorrhage: a population-based case-control study in Australasia.
A healthy, balanced diet can prevent stroke, but little is known about dietary risk factors for subarachnoid hemorrhage (SAH). We aimed to determine the relationship between common dietary habits and risk of SAH. In a population-based, case-control study of SAH undertaken across 4 Australasian cities, a standardized questionnaire was used to obtain information on the frequency of consumption of 15 common food items and alcohol in incident cases (n = 383) and frequency-matched community controls (n = 473). Logistic regression models were used to estimate the independent effects of these dietary factors, after adjusting for conventional risk factors for SAH. Data are reported with odds ratios (OR) and 95% confidence intervals (CI). The risk of SAH rose with increasing consumption of fat or skin on meat (p trend = 0.04), being highest in those with consumption >4 times weekly compared with no fat or skin on meat (adjusted OR 1.70, 95% CI 1.09-2.66), while use of skim or reduced-fat milk (p trend = 0.01) and fruit (p trend = 0.04) was associated with a reduced risk of SAH compared with rare use. Among people with a history of hypertension, frequently adding salt to food was associated with an increased risk of SAH, irrespective of whether they were (OR 2.58, 95% CI 1.29-5.13) or were not (OR 2.88, 95% CI 1.46-5.70) currently taking antihypertensive treatment. Frequent intake of fat appears to be associated with an increased risk of SAH, particularly in people with hypertension, while frequent use of skim or reduced-fat milk and fruit appears protective for SAH.


Decoupling of N-acetyl-aspartate and glutamate within the dorsolateral prefrontal cortex in schizophrenia.
Aberrant function of glutamatergic pathways is likely to underlie the pathology of schizophrenia. Evidence of oxidative stress in the disease pathology has also been reported. N-Acetylaspartate (NAA) is metabolically linked to both cascades and may be a key marker in exploring the interconnection of glutamatergic pathways and oxidative stress. Several studies have reported positive correlation between the levels of NAA and Glx (the sum of glutamate and glutamine) in several brain regions in healthy subjects, by using proton magnetic resonance spectroscopy ([(1)H]MRS). Interestingly, one research group recently reported decoupling of the relationship between NAA and Glx in the hippocampus of patients with schizophrenia. Here we report levels of NAA and Glx measured using [(1)H]MRS, relative to the level of creatine (Cr) as an internal control. The dorsolateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC) in 25 patients with schizophrenia and 17 matched healthy controls were studied. In DLPFC, NAA/Cr and Glx/Cr were significantly positively correlated in healthy controls after correction for the effect of age and smoking status and after correction for multiple comparisons (r= 0.627, P= 0.017). However, in patients with schizophrenia, the positive correlation between NAA/Cr and Glx/Cr was not observed even after correcting for these two variables (r= -0.330, P= 0.124). Positive correlation between NAA/Cr and Glx/Cr was not observed in the ACC in both groups. Decoupling of NAA and Glx in the DLPFC may reflect the interconnection of glutamatergic pathways and oxidative stress in the pathology of schizophrenia, and may possibly be a biomarker of the disease.


Treatment of complex idiopathic clubfoot using the modified Ponseti method: up to 11 years follow-up.
Complex idiopathic clubfeet are distinguished by significant shortening, rigid equinus with a deep crease above the heel, severe plantar flexion of all metatarsals, a deep plantar crease seven across the full width of the sole of the foot and high cavus with a short and hyperextended big toe. Ponseti has devised a modified technique for treating complex clubfeet. We retrospectively identified 11 children (nine males and two females) with 17 complex clubfeet who were treated with the modified Ponseti method. Demographics, severity of clubfoot, number of casts, rate of tendoachilles tenotomy, relapse rate and their management, any additional procedures and data on complications were collected. The average follow-up was 7 years (range 3-11 years) and the average Pirani score was 5.5 (range 4.5-6.0). Initial correction was achieved in all children, with an average of 7 (range 5-10) Ponseti casts. Tendoachilles tenotomy was performed in all 17 feet (100%). The overall relapse rate was 53% (nine feet). Five relapses were managed successfully with repeat casting and four feet were subjected to a second tendoachilles tenotomy. Four feet required extensive surgical releases. A satisfactory outcome was achieved at the final follow-up in 13 of 17 feet (76.5%). Two of these children (two feet) required an additional tibialis anterior transfer. In our experience, the modified Ponseti method is an effective first-line treatment for complex idiopathic clubfoot; however, such children will often require more casts than usual and have a higher rate of tendoachilles tenotomy and a higher risk of relapse requiring surgical procedures. level IV.