Functional Classification of the Medial Ulnar Collateral Ligament: An In Vivo Kinematic Study With Computer-Aided Design.
It has been widely accepted that the anterior and posterior bundles of the medial ulnar collateral ligament (MUCL) tighten at extension and flexion, respectively. However, this belief is based on anatomic data acquired from cadaveric studies. The advancement of 3-dimensional (3D) model technology has made possible the simulation of dynamic movement that includes each ligament bundle fiber to analyze its functional properties. To date, no study has analyzed ligament kinematics at the level of the fibers while also focusing on their functional properties. To propose a new classification for functional properties of the MUCL based on its kinematic pattern. Descriptive laboratory study. Five healthy elbow joints were scanned by use of computed tomography, and 3D models were rendered and translated into vertex points for further mathematical analysis. The humeral origin and ulnar insertion of the MUCL fiber groups were registered. Each vertex point on the origin side was randomly connected to the insertion side, with each pair of corresponding points defined as 1 ligament fiber. Lengths of all the fibers were measured at 1° increments of elbow range of motion (ROM). Ligament fibers were grouped according to their patterns. Mean coverage area for each group, expressed as the percentage of ligament fibers per group to the total number of fibers, was calculated. Four major bundle groups were found based on fiber length properties. Kinematic simulation showed that each group had a different kinematic function throughout elbow ROM. Mean coverage area of groups 1, 2, 3, and 4 was 8% ± 4%, 10% ± 5%, 42% ± 6%, and 40% ± 8%, respectively. Each group acted as a dominant stabilizer in certain arcs of motion. Reciprocal activity was observed between groups 1 and 3 along with groups 2 and 4 to produce synergistic properties of maintaining elbow stability. Detailed analysis of fibers of the MUCL allows for further understanding of its kinematic function. This study provides MUCL group coverage area and kinematic function for each degree of motion arc, allowing selective reconstruction of the MUCL according to mechanism of injury. Understanding the dominant functional fibers of the MUCL will benefit surgeons attempting MUCL reconstruction and will enhance further anatomic study.


Safety and immunogenicity of the candidate tuberculosis vaccine MVA85A in West Africa.
Vaccination with a recombinant modified vaccinia Ankara expressing antigen 85A from Mycobacterium tuberculosis, MVA85A, induces high levels of cellular immune responses in UK volunteers. We assessed the safety and immunogenicity of this new vaccine in West African volunteers. We vaccinated 21 healthy adult male subjects (11 BCG scar negative and 10 BCG scar positive) with MVA85A after screening for evidence of prior exposure to mycobacteria. We monitored them over six months, observing for clinical, haematological and biochemical adverse events, together with assessment of the vaccine induced cellular immune response using ELISPOT and flow cytometry. MVA85A was well tolerated with no significant adverse events. Mild local and systemic adverse events were consistent with previous UK trials. Marked immunogenicity was found whether individuals had a previous BCG scar or not. There was not enhanced immunogenicity in those with a BCG scar, and induced T cell responses were better maintained in apparently BCG-naïve Gambians than previously studied BCG-naïve UK vaccinees. Although responses were predominantly attributable to CD4+ T cells, we also identified antigen specific CD8+ T cell responses, in subjects who were HLA B-35 and in whom enough blood was available for more detailed immunological analysis. These data on the safety and immunogenicity of MVA85A in West Africa support its accelerated development as a promising booster vaccine for tuberculosis. ClinicalTrials.gov NCT00423839.


Differentiation of murine erythroleukemia cells results in the rapid repression of vimentin gene expression.
We show that vimentin filaments are present in undifferentiated Friend murine erythroleukemia cells, but are lost progressively to undetectable levels by 96 h of dimethyl sulfoxide-mediated differentiation. The amount of newly synthesized cytoskeletal vimentin is decreased dramatically by 24 h of induction, and is paralleled by a rapid loss of vimentin mRNA (approximately 25-fold reduction at 96 h). Hence, disappearance of vimentin filaments in these cells appears to be regulated at the level of vimentin mRNA abundance. On the other hand, the levels of actin synthesis and actin mRNA remain essentially unchanged. The kinetics of vimentin mRNA reduction during dimethyl sulfoxide-mediated differentiation, and the levels of vimentin mRNA observed in the presence of hexamethylene-bisacetamide or hemin as inducers suggest that the cessation of vimentin expression precedes, but may be associated with commitment to terminal differentiation. Our results demonstrate the dynamic regulation of vimentin expression in mammalian erythropoiesis.


Are the school version of the assessment of motor and process skills measures valid for German-speaking children?
There are no validated assessment tools for evaluating quality of schoolwork task performance of children living in German-speaking Europe (GSE). To determine whether the international age-normative means of the School Version of the Assessment of Motor and Process Skills (School AMPS) are valid for use in GSE. The participants were 159 typically-developing children, 3-12 years, from GSE. We examined the proportions of School AMPS measures falling within ±2 standard deviation (SD) of the international age-normative means, and evaluated for significant group differences (p < 0.05) in mean School AMPS measures between the GSE sample and the international age-normative sample using one-sample Z tests. When significant mean differences were found, we evaluated if the differences were clinically meaningful. At least 95% of the GSE School AMPS measures fell within ±2 SD of the international age-normative means for the School AMPS. The only significant mean differences were for 6- (p < 0.01) and 8-year-olds (p = 0.02), and only the 6-year-old school process mean difference was clinically meaningful. Because the only identified clinically meaningful difference was associated with likely scoring error of one rater, the international age-normative means of the School AMPS appear to be valid for use with children in GSE.


Filipino children exhibit catch-Up growth from age 2 to 12 years.
Potential for catch-up growth among stunted children is thought to be limited after age 2 y, particularly when they remain in poor environments. We explored the extent to which there were improvements in height status from age 2 to 12 y in a cohort of >2, 000 children from the Cebu (The Philippines) Longitudinal Health and Nutrition Survey. At age 2 y, about 63% of sample children were stunted as defined by height-for-age (HAZ) <-2 based on the WHO reference. Of children stunted at age 2, 30% were no longer stunted at 8.5 y, and 32.5% were no longer stunted at 12 y. The mean increase in HAZ among those with such improvements was 1.14 units. The likelihood that children stunted at age 2 y would no longer be stunted at 8.5 y was estimated using logistic regression. Low birth weight, which was associated with more severe stunting in the first 2 y of life, significantly reduced likelihood of catch-up growth in later childhood. In contrast, children with taller mothers, who were first born, longer at birth, less severely stunted in early infancy and those with fewer siblings were more likely to increase HAZ from <-2 to >-2 between ages 2 and 8.5 y. Similar factors predicted the improvement in linear growth from 8.5 to 12 y. These results suggest that there is a large potential for catch-up growth in children into the preadolescent years.


What is inverse-geometry CT?
Inverse-geometry computed tomography (IGCT) systems are being developed to provide improved volumetric imaging. In conventional multislice CT systems, x-rays are emitted from a small area and irradiate a large-area detector. In an IGCT system, x-ray sources are distributed over a large area, with each beam irradiating a small-area detector. Therefore, in the inverse geometry, a series of narrow x-ray beams are switched on and off while the gantry rotates. In conventional CT geometry, cone-beam and scatter artifacts increase with the imaged volume thickness. An inverse geometry may be less susceptible to scatter effects, because only a fraction of the field of view is irradiated at one time. The distributed source in the inverse geometry potentially improves sampling, leading to reduced cone-beam artifacts. In the inverse geometry, the tube current may be adjusted separately for each source location, which potentially reduces dose. Multiple IGCT prototypes have been constructed and tested on phantoms. A gantry-based IGCT system with one-second gantry rotation was developed, and images of phantoms and small animals were successfully acquired. Clinical feasibility with acceptable noise levels and scan times has not yet been shown. Overall, results from prototype systems suggest that the inverse geometry will enable imaging of a thick volume (∼16 cm) while potentially reducing cone-beam artifacts, scatter effects, and radiation dose. The magnitude of these benefits will depend on the specific IGCT implementation and need to be quantified relative to comparable multislice scanners.


Targeting the postpartum period to promote weight loss: a systematic review and meta-analysis.
Many international clinical guidelines recommend that overweight and obese women lose weight prior to pregnancy to reduce the risk of adverse pregnancy outcomes. Women who have recently given birth and plan future pregnancies are an important target population for preconception weight-loss interventions. A systematic review to evaluate postpartum dietary and/or physical activity interventions to promote weight loss and improve health in a subsequent pregnancy was conducted. Five databases-the Cochrane Central Register of Controlled Trials, MEDLINE (through PubMed), Embase, the Australian New Zealand Clinical Trials Registry, and the International Clinical Trials Registry-were searched using the following terms: preconception, pregnancy, postpartum, pregnancy outcomes, body mass index, weight gain, weight loss, weight change, postpartum weight retention, dietary or lifestyle intervention, and randomiz(s)ed controlled trial. The date of last search was November 2017. Data were extracted from each identified study using a standard form. The primary outcomes were weight loss at the completion of the intervention and at follow-up assessments. Secondary endpoints included maternal and infant outcomes in a subsequent pregnancy. Mean differences (MDs) were calculated for continuous data and risk ratios for dichotomous data, both with 95%CIs. A total of 235 abstracts (193 after duplicates were excluded) were identified, from which 37 manuscripts were selected for full-text review. In total, 27 trials were identified for inclusion. Outcome data were available for approximately 75% of participants (n = 3485). A combined dietary and physical activity intervention provided post partum produced greater postpartum weight loss (MD, -2.49 kg; 95%CI, -3.34 to -1.63 kg [random-effects model]; 12 studies, 1156 women), which was maintained at 12 months post partum (MD, -2.41 kg; 95%CI, -3.89 to -0.93 kg [random-effects model]; 4 studies, 405 women), compared with no intervention. No studies reported maternal or infant health outcomes in a subsequent pregnancy. Providing a postpartum intervention is associated with weight loss after birth, but effects on maternal and infant health in a subsequent pregnancy are uncertain.


Radiation-resistant and radiation-sensitive forms of host resistance to polyomavirus.
Newborn mice of several inbred strains develop few or no tumors following inoculation with highly tumorigenic strains of polyomavirus. Here we show that such resistant strains can be divided into two groups based on the responses of adult mice to radiation followed by virus inoculation. Most strains show a radiation-sensitive form of resistance (Rr-s) and develop tumors following radiation and virus challenge. This type of resistance has previously been recognized as immunological, based on T-cell responses against virus-encoded neoantigen(s) expressed in tumor cells. Other strains exhibit a radiation-resistant form of resistance (Rr-r) and fail to develop tumors when treated in the same manner. Three additional properties of Rr-r mice distinguish them from Rr-s mice: (i) survival of newborns following inoculation with a highly virulent and usually lethal strain of virus, (ii) resistance to virus spread in newborns inoculated with either tumorigenic or virulent virus strains, and (iii) dominant or semidominant transmission of resistance in crosses with a highly susceptible strain. The Rr-r phenotype reflects a constitutive nonimmunological type of resistance that is targeted to the virus and blocks its dissemination.


Enhanced sensitivity to cisplatin and gemcitabine in Brca1-deficient murine mammary epithelial cells.
Breast cancers due to germline mutations or altered expression of the BRCA1 gene associate with an aggressive clinical course and frequently exhibit a "triple-negative" phenotype, i.e. lack of expression of the estrogen and progesterone hormone receptors and lack of overexpression of the HER2/NEU oncogene, thereby rendering them relatively insensitive to hormonal manipulation and targeted HER2 therapy, respectively. BRCA1 plays a role in multiple DNA repair pathways, and thus, when mutated, results in sensitivity to certain DNA damaging drugs. Here, we used a Brca1 murine mammary epithelial cell (MMEC) model to examine the effect of loss of Brca1 on cellular sensitivity to various chemotherapy drugs. To explore novel therapeutic strategies, we included DNA damaging and non-DNA damaging drugs whose mechanisms are dependent and independent of DNA repair, respectively, and drugs that are used in standard and non-standard lines of therapy for breast cancer. To understand the cellular mechanism, we also determined the role that DNA repair plays in sensitivity to these drugs. We found that cisplatin and gemcitabine had the greatest specific therapeutic benefit to Brca1-deficient MMECs, and that when used in combination produced a synergistic effect. This sensitivity may be attributed in part to defective NER, which is one of the DNA repair pathways normally responsible for repairing DNA adducts produced by cisplatin and is shown in this study to be defective in Brca1-deficient MMECs. Brca1-deficient MMECs were not differentially sensitive to the standard breast cancer chemotherapy drugs doxorubicin, docetaxel or 5-FU. Both cisplatin and gemcitabine should be explored in clinical trials for first line regimens for BRCA1-associated and triple-negative breast cancer.


Viral suppression rates in salvage treatment with raltegravir improved with the administration of genotypic partially active or inactive nucleoside/tide reverse transcriptase inhibitors.
Nucleoside reverse transcriptase inhibitors (NRTIs) are often administered in salvage therapy even if genotypic resistance tests (GRTs) indicate high-level resistance, but little is known about the benefit of these additional NRTIs. The effect of <2 compared with 2 NRTIs on viral suppression (HIV-1 RNA < 50 copies/mL) at week 24 was studied in salvage patients receiving raltegravir. Intent-to-treat and per-protocol analyses were performed; last observation carried forward imputation was used to deal with missing information. Logistic regressions were weighted to create a pseudopopulation in which the probability of receiving <2 and 2 NRTIs was unrelated to baseline factors predicting treatment response. One-hundred thirty patients were included, of whom 58.5% (n = 76) received <2 NRTIs. NRTIs were often replaced by other drug classes. Patients with 2 NRTIs received less additional drug classes compared with patients with <2 NRTIs [median (IQR): 1 (1-2) compared with 2 (1-2), P Wilcoxon < 0.001]. The activity of non-NRTI treatment components was lower in the 2 NRTIs group compared with the <2 NRTIs group [median (IQR) genotypic sensitivity score: 2 (1.5-2.5) compared with 2.5 (2-3), P Wilcoxon < 0.001]. The administration of <2 NRTIs was associated with a worse viral suppression rate at week 24. The odds ratios were 0.34 (95% confidence interval: 0.13 to 0.89, P = 0.027) and 0.19 (95% confidence interval: 0.05 to 0.79, P = 0.023) when performing the last observation carried forward and the per-protocol approach, respectively. Our findings showed that partially active or inactive NRTIs contribute to treatment response, and thus the use of 2 NRTIs in salvage regimens that include raltegravir seems warranted.


Molecular dynamics simulations and membrane protein structure quality.
Despite a growing repertoire of membrane protein structures (currently approximately 120 unique structures), considerations of low resolution and crystallization in the absence of a lipid bilayer require the development of techniques to assess the global quality of membrane protein folds. This is also the case for assessment of, e.g. homology models of human membrane proteins based on structures of (distant) bacterial homologues. Molecular dynamics (MD) simulations may be used to help evaluate the quality of a membrane protein structure or model. We have used a structure of the bacterial ABC transporter MsbA which has the correct transmembrane helices but an incorrect handedness and topology of their packing to test simulation methods of quality assessment. An MD simulation of the MsbA model in a lipid bilayer is compared to a simulation of another bacterial ABC transporter, BtuCD. The latter structure has demonstrated good conformational stability in the same bilayer environment and over the same timescale (20 ns) as for the MsbA model simulation. A number of comparative analyses of the two simulations were performed to assess changes in the structural integrity of each protein. The results show a significant difference between the two simulations, chiefly due to the dramatic structural deformations of MsbA. We therefore propose that MD could become a useful quality control tool for membrane protein structural biology. In particular, it provides a way in which to explore the global conformational stability of a model membrane protein fold.


Effects of aging and estradiol supplementation on GH axis dynamics in women.
GH and IGF-I secretion decrease with age. The decline in serum GH with age appears to be associated with menopause. Prior studies of GH release before and after oral and transdermal hormonal replacement in the postmenopausal patient have shown no change or an increase in GH secretion. To distinguish the somatotropic axis effects of aging from those of estrogen deficiency, we compared eight prematurely menopausal women, aged 25-40 yr, with eight postmenopausal women, aged 51-70 yr, both before and after estradiol replacement. All women had a body mass index below 28 kg/m2. All were evaluated twice with frequent blood sampling every 10 min for 24 h. Studies were performed in the absence of exogenous hormones and 6-8 wk after transdermal estradiol replacement, targeted to achieve a serum estradiol level of 367 pmol/liter. GH pulsatility was analyzed. Variables tested included mean GH levels, interpulse baseline mean, pulse frequency per 24 h, and pulse amplitude. Transdermal estrogen replacement had a significant effect on mean GH levels and mean basal GH levels in both the premature ovarian failure and the age-appropriate postmenopausal group. No differences were noted in GH pulse frequency, GH pulse amplitude, IGF-I, IGF-binding protein-1, and IGF-binding protein-3 before and after treatment. A pronounced age effect was noted between the two groups. The premature ovarian failure women secreted significantly greater mean GH than the age-appropriate postmenopausal group regardless of treatment, with a significance level of P = 0.026. Interpulse baseline GH means were greater in the premature ovarian failure women than in the age-appropriate postmenopausal group, but the significance of this relationship was obliterated after adjustment for body mass index. Pulse amplitude was significantly increased in the premature ovarian failure women compared with age-appropriate postmenopausal women (P = 0.006). No significant changes were detected in the GH pulse frequency between the premature ovarian failure and postmenopausal groups. We conclude that moderate doses of transdermal estradiol supplementation do not exert a great effect on the somatotropic axis in women. Age and body composition appear to be the predominant influences on GH activity in women.


Comparison of body habitus in patients with pulmonary arterial hypertension enrolled in the Registry to Evaluate Early and Long-term PAH Disease Management with normative values from the National Health and Nutrition Examination Survey.
To investigate the correlation between body mass index (BMI) and pulmonary artery systolic pressure in a large population of patients with pulmonary arterial hypertension (PAH). The BMI of patients with group 1 PAH enrolled in the Registry to Evaluate Early and Long-term PAH Disease Management (REVEAL) was compared with that of age- and sex-matched controls in the National Health and Nutrition Examination Survey (NHANES) to clarify whether obesity is linked with PAH. The diagnosis of PAH was defined in REVEAL by right-sided heart catheterization. Differences in BMI and the percentage of patients considered obese (BMI ≥30) and underweight (BMI <18.5) in various subgroups of patients enrolled in REVEAL from March 30, 2006, through September 11, 2007, were determined. Mean BMI was no different for patients with PAH (n=2141) than for the NHANES normal comparison group; however, the proportion of obese and underweight patients was increased in patients with PAH. Subgroup analysis demonstrated that subgroups with idiopathic PAH and those with PAH associated with drugs and toxins had both higher BMI and percentage of obese patients, whereas 3 other subgroups (those with PAH associated with congenital heart disease, connective tissue disease, and human immunodeficiency virus) had lower mean BMI. Mean BMI of the REVEAL patients was the same as that of the NHANES normal comparison group; however, there were higher percentages of obese and underweight patients in REVEAL. This discrepancy can be explained by the balancing effect of more overweight and underweight patients in different PAH subgroups. The reason for the increased frequency of obesity in idiopathic PAH is unknown, and additional study is needed.


Inhibitory effect of breast milk on infectivity of live oral rotavirus vaccines.
Live oral rotavirus vaccines have been less immunogenic and efficacious among children in poor developing countries compared with middle income and industrialized countries for reasons that are not yet completely understood. We assessed whether the neutralizing activity of breast milk could lower the titer of vaccine virus and explain this difference in vitro. Breast milk samples were collected from mothers who were breast-feeding infants 4 to 29 weeks of age (ie, vaccine eligible age) in India (N = 40), Vietnam (N = 77), South Korea (N = 34), and the United States (N = 51). We examined breast milk for rotavirus-specific IgA and neutralizing activity against 3 rotavirus vaccine strains-RV1, RV5 G1, and 116E using enzyme immunoassays. The inhibitory effect of breast milk on RV1 was further examined by a plaque reduction assay. Breast milk from Indian women had the highest IgA and neutralizing titers against all 3 vaccine strains, while lower but comparable median IgA and neutralizing titers were detected in breast milk from Korean and Vietnamese women, and the lowest titers were seen in American women. Neutralizing activity was greatest against the 2 vaccine strains of human origin, RV1 and 116E. This neutralizing activity in one half of the breast milk specimens from Indian women could reduce the effective titer of RV1 by ∼2 logs, of 116E by 1.5 logs, and RV5 G1 strain by ∼1 log more than that of breast milk from American women. The lower immunogenicity and efficacy of rotavirus vaccines in poor developing countries could be explained, in part, by higher titers of IgA and neutralizing activity in breast milk consumed by their infants at the time of immunization that could effectively reduce the potency of the vaccine. Strategies to overcome this negative effect, such as delaying breast-feeding at the time of immunization, should be evaluated.


The roles of oxidized low-density lipoprotein and interleukin-6 levels in acute atherothrombotic and lacunar ischemic stroke.
The role of circulating, oxidized low-density lipoprotein and interleukin-6 levels in acute ischemic stroke considering the primary-vessel disease was investigated. The study consisted of 28 patients with acute ischemic stroke and 23 control subjects. Patients were subdivided into large-vessel (n = 12) and small-vessel (n =16) disease stroke groups according to the Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria. The means of oxidized low-density lipoprotein and interleukin-6 levels of patients with acute ischemic stroke were higher than controls (P < .01, P < .05). Mean oxidized low-density lipoprotein level was higher in the large-vessel disease group than in the small-vessel disease group (P < .01). The mean of inteleukin-6 levels was higher in the small-vessel disease group (P < .01). The results of the present study showed that oxidative stress promotes large-vessel disease rather than small-vessel disease stroke, and inflammation may play important an role in the development of small-vessel disease stroke.


[Our experience with the Nesbit technique for the treatment of Peyronie's disease].
The Nesbit technique is the reference technique for the surgical correction of the penile incurvation of Peyronie's disease. We report our experience at the Centro Urológico San Ignacio. 68 patients with stable penile incurvation who requested surgical correction. Correction of the curvature is defined as > or = 80% rectification of the pathologic angle. Mean patient age was 44 years (31-77). 53% of the patients presented difficulties for penetration, 20% partner's discomfort on intercourse, and 27% requested surgery for aesthetic reasons. Penile deviation was dorsal in 42% of the patients, to the left in 46% and to the right in 12%. The curvature angle was less than 30 degrees in 28%, between 30 degrees-45 degrees in 36%, between 45 degrees-60 degrees in 37% and > 60 degrees in 7%. Mean follow-up of the series was 36 months. 20% of the patients had a penile shortening longer than 1.5 cm. 85% of the patients are satisfied with the results. The most frequent complaint was penile shortening. Correction was achieved in 92% of the patients. Four patients were reoperated: two of them for significant phimosis three months after surgery, and the other two for curvature recurrence. The Nesbit technique is a simple technique, with a low complication rate, and good results both in curvature correction and patient satisfaction.


Consensus Paper: Ataxic Gait.
The aim of this consensus paper is to discuss the roles of the cerebellum in human gait, as well as its assessment and therapy. Cerebellar vermis is critical for postural control. The cerebellum ensures the mapping of sensory information into temporally relevant motor commands. Mental imagery of gait involves intrinsically connected fronto-parietal networks comprising the cerebellum. Muscular activities in cerebellar patients show impaired timing of discharges, affecting the patterning of the synergies subserving locomotion. Ataxia of stance/gait is amongst the first cerebellar deficits in cerebellar disorders such as degenerative ataxias and is a disabling symptom with a high risk of falls. Prolonged discharges and increased muscle coactivation may be related to compensatory mechanisms and enhanced body sway, respectively. Essential tremor is frequently associated with mild gait ataxia. There is growing evidence for an important role of the cerebellar cortex in the pathogenesis of essential tremor. In multiple sclerosis, balance and gait are affected due to cerebellar and spinal cord involvement, as a result of disseminated demyelination and neurodegeneration impairing proprioception. In orthostatic tremor, patients often show mild-to-moderate limb and gait ataxia. The tremor generator is likely located in the posterior fossa. Tandem gait is impaired in the early stages of cerebellar disorders and may be particularly useful in the evaluation of pre-ataxic stages of progressive ataxias. Impaired inter-joint coordination and enhanced variability of gait temporal and kinetic parameters can be grasped by wearable devices such as accelerometers. Kinect is a promising low cost technology to obtain reliable measurements and remote assessments of gait. Deep learning methods are being developed in order to help clinicians in the diagnosis and decision-making process. Locomotor adaptation is impaired in cerebellar patients. Coordinative training aims to improve the coordinative strategy and foot placements across strides, cerebellar patients benefiting from intense rehabilitation therapies. Robotic training is a promising approach to complement conventional rehabilitation and neuromodulation of the cerebellum. Wearable dynamic orthoses represent a potential aid to assist gait. The panel of experts agree that the understanding of the cerebellar contribution to gait control will lead to a better management of cerebellar ataxias in general and will likely contribute to use gait parameters as robust biomarkers of future clinical trials.


Association of peripartum management and high maternal blood loss at cesarean delivery for placenta accreta spectrum (PAS): A multinational database study.
Placenta accreta spectrum (PAS) carries a high burden of adverse maternal outcomes, especially significant blood loss, which can be life-threatening. Different management strategies have been proposed but the association of clinical risk factors and surgical management options during cesarean delivery with high blood loss is not clear. In this international multicenter study, 338 women with PAS undergoing cesarean delivery were included. Fourteen European and one non-European center (USA) provided cases treated retrospectively between 2008 and 2014 and prospectively from 2014 to 2019. Peripartum blood loss was estimated visually and/or by weighing and measuring of volume. Participants were grouped based on blood loss above or below the 75th percentile (>3500 ml) and the 90th percentile (>5500 ml). Placenta percreta was found in 58% of cases. Median blood loss was 2000 ml (range: 150-20 000 ml). Unplanned hysterectomy was associated with an increased risk of blood loss >3500 ml when compared with planned hysterectomy (adjusted OR [aOR] 3.7 [1.5-9.4], p = 0.01). Focal resection was associated with blood loss comparable to that of planned hysterectomy (crude OR 0.7 [0.2-2.1], p = 0.49). Blood loss >3500 ml was less common in patients undergoing successful conservative management (placenta left in situ, aOR 0.1 [0.0-0.6], p = 0.02) but was more common in patients who required delayed hysterectomy (aOR 6.5 [1.7-24.4], p = 0.001). Arterial occlusion methods (uterine or iliac artery ligation, embolization or intravascular balloons), application of uterotonic medication or tranexamic acid showed no significant effect on blood loss >3500 ml. Patients delivered by surgeons without experience in PAS were more likely to experience blood loss >3500 ml (aOR 3.0 [1.4-6.4], p = 0.01). In pregnant women with PAS, the likelihood of blood loss >3500 ml was reduced in planned vs unplanned cesarean delivery, and when the surgery was performed by a specialist experienced in the management of PAS. This reinforces the necessity of delivery by an expert team. Conservative management was also associated with less blood loss, but only if successful. Therefore, careful patient selection is of great importance. Our study showed no consistent benefit of other adjunct measures such as arterial occlusion techniques, uterotonics or tranexamic acid.


The Impact of Pharmacokinetic Interactions With Eslicarbazepine Acetate Versus Oxcarbazepine and Carbamazepine in Clinical Practice.
Eslicarbazepine acetate (ESL) is a new anti-epileptic drug (AED) chemically related to oxcarbazepine (OXC) and carbamazepine (CBZ) and is increasingly used in clinical practice. The purpose of the study was to investigate 2-way pharmacokinetic interactions between ESL and other AEDs as compared to OXC and CBZ. Anonymous data regarding age, gender, use of AEDs, daily doses and serum concentration measurements of ESL, OXC, CBZ and lamotrigine (LTG) and other AEDs were retrieved from 2 therapeutic drug monitoring (TDM) databases in Norway. Drugs were categorized according to their known potential for interactions. Concentration/dose (C/D) ratios were calculated. Data from 1100 patients were available. The C/D ratios of ESL and OXC were unchanged in combination with enzyme-inducing AEDs or valproate (VPA). The C/D ratio of CBZ decreased by 40% and 22% in combination with other enzyme-inducing AEDs or VPA, respectively, pointing to an increased clearance. ESL demonstrated no significant enzyme-inducing effect on LTG metabolism although there was a 20% and 34% decrease in the C/D ratio of LTG in combination with OXC and CBZ, respectively. Possible pharmacokinetic interactions have been studied for ESL as compared to OXC and CBZ. The pharmacokinetics of ESL is not affected by enzyme-inducing AEDs or VPA and does not affect the metabolism of LTG in contrast to OXC and CBZ. The study demonstrates the value of using TDM databases to explore the potential for pharmacokinetic interactions of new AEDs.


Evolution of the DAN gene family in vertebrates.
The DAN gene family (DAN, Differential screening-selected gene Aberrant in Neuroblastoma) is a group of genes that is expressed during development and plays fundamental roles in limb bud formation and digitation, kidney formation and morphogenesis and left-right axis specification. During adulthood the expression of these genes are associated with diseases, including cancer. Although most of the attention to this group of genes has been dedicated to understanding its role in physiology and development, its evolutionary history remains poorly understood. Thus, the goal of this study is to investigate the evolutionary history of the DAN gene family in vertebrates, with the objective of complementing the already abundant physiological information with an evolutionary context. Our results recovered the monophyly of all DAN gene family members and divide them into five main groups. In addition to the well-known DAN genes, our phylogenetic results revealed the presence of two new DAN gene lineages; one is only retained in cephalochordates, whereas the other one (GREM3) was only identified in cartilaginous fish, holostean fish, and coelacanth. According to the phyletic distribution of the genes, the ancestor of gnathostomes possessed a repertoire of eight DAN genes, and during the radiation of the group GREM1, GREM2, SOST, SOSTDC1, and NBL1 were retained in all major groups, whereas, GREM3, CER1, and DAND5 were differentially lost.


Biofilm formation and sanitizer resistance of Escherichia coli O157:H7 strains isolated from "high event period" meat contamination.
In the meat industry, a "high event period" (HEP) is defined as a time period during which commercial meat plants experience a higher than usual rate of Escherichia coli O157:H7 contamination. Genetic analysis indicated that within a HEP, most of the E. coli O157:H7 strains belong to a singular dominant strain type. This was in disagreement with the current beef contamination model stating that contamination occurs when incoming pathogen load on animal hides, which consists of diverse strain types of E. coli O157:H7, exceeds the intervention capacity. Thus, we hypothesize that the HEP contamination may be due to certain in-plant colonized E. coli O157:H7 strains that are better able to survive sanitization through biofilm formation. To test our hypothesis, a collection of 45 E. coli O157:H7 strains isolated from HEP beef contamination incidents and a panel of 47 E. coli O157:H7 strains of diverse genetic backgrounds were compared for biofilm formation and sanitizer resistance. Biofilm formation was tested on 96-well polystyrene plates for 1 to 6 days. Biofilm cell survival and recovery growth after sanitization were compared between the two strain collections using common sanitizers, including quaternary ammonium chloride, chlorine, and sodium chlorite. No difference in "early stage" biofilms was observed between the two strain collections after incubation at 22 to 25°C for 1 or 2 days. However, the HEP strains demonstrated significantly higher potency of "mature" biofilm formation after incubation for 4 to 6 days. Biofilms of the HEP strains also exhibited significantly stronger resistance to sanitization. These data suggest that biofilm formation and sanitization resistance could have a role in HEP beef contamination by E. coli O157:H7, which highlights the importance of proper and complete sanitization of food contact surfaces and food processing equipment in commercial meat plants.


The Drosophila inhibitor of apoptosis protein DIAP2 functions in innate immunity and is essential to resist gram-negative bacterial infection.
The founding member of the inhibitor of apoptosis protein (IAP) family was originally identified as a cell death inhibitor. However, recent evidence suggests that IAPs are multifunctional signaling devices that influence diverse biological processes. To investigate the in vivo function of Drosophila melanogaster IAP2, we have generated diap2 null alleles. diap2 mutant animals develop normally and are fully viable, suggesting that diap2 is dispensable for proper development. However, these animals were acutely sensitive to infection by gram-negative bacteria. In Drosophila, infection by gram-negative bacteria triggers the innate immune response by activating the immune deficiency (imd) signaling cascade, a NF-kappaB-dependent pathway that shares striking similarities with the pathway of mammalian tumor necrosis factor receptor 1 (TNFR1). diap2 mutant flies failed to activate NF-kappaB-mediated expression of antibacterial peptide genes and, consequently, rapidly succumbed to bacterial infection. Our genetic epistasis analysis places diap2 downstream of or in parallel to imd, Dredd, Tak1, and Relish. Therefore, DIAP2 functions in the host immune response to gram-negative bacteria. In contrast, we find that the Drosophila TNFR-associated factor (Traf) family member Traf2 is dispensable in resistance to gram-negative bacterial infection. Taken together, our genetic data identify DIAP2 as an essential component of the Imd signaling cascade, protecting the organism from infiltrating microbes.


Meshwork structures in bovine glomerular and tubular basement membrane as revealed by ultra-high-resolution scanning electron microscopy.
We examined the ultrastructure of the bovine glomerular basement membrane (GBM) and tubular basement membrane (TBM) using ultra-high-resolution scanning electron microscopy after conductive staining without metal coating. Purified basement membranes (BMs) were obtained by sonication and acellular BMs by detergent treatments. Purified GBMs (PGBMs) and acellular GBMs (AGBMs) showed similar meshwork structures composed of regular round or oval pores and branching strands. Pore diameters were 10.2 +/- 2.4 nm (mean +/- SD) in PGBMs and 9.8 +/- 3.1 nm in AGBMs. Purified TBMs (PTBMs) and acellular TBMs (ATBMs) exhibited heterogeneous meshwork structures in which compact meshes in the cristae were combined with coarse meshes in the invaginations on the epithelial surfaces. Pore diameters were 12.6 +/- 5.2 nm in PTBMs and 11.4 +/- 4.0 nm in ATBMs which were significantly larger than those in GBMs. The width of the strands ranged from 3 to 15 nm in all BMs. A 4 M guanidine hydrochloride extraction of the acellular BMs revealed large polygonal networks in the invaginations of the TBM and twisted strands which were considered to be type IV collagen fibrils. Ultra-high-resolution scanning electron microscopy and conductive staining were useful for the study of three-dimensional architectures of BMs and revealed heterogeneous meshwork structures in the GBM and TBM which were probably caused by a different ratio of the major components.


Midgut neuroendocrine tumours with liver metastases: results of the UKINETS study.
We intended to identify the prognostic factors and the results of interventions on patients with liver metastatic midgut carcinoids. Five institutions that are part of United Kingdom and Ireland neuroendocrine tumour (NET) group took part in this study. Patients were included if they had histology proven NET of midgut origin and liver metastases at the time of the study. Clinical and biochemical data were collected retrospectively from hospital charts, pathology reports, radiology reports and biochemistry records for each patient. Three hundred and sixty patients were included in the study. The median survival from date of diagnosis was 7.69 years (confidence interval (CI) 6.40-8.99) and 5.95 years (CI 5.02-6.88) from date of diagnosis of liver metastases. On univariate analysis, increasing age at diagnosis, increasing urinary hydroxyindole acetic acid levels, increasing plasma chromogranin A levels, high Ki67, high tumour volume and treatment with chemotherapy were identified as factors associated with a significantly poorer outcome. Resection of liver metastases, resection of small bowel primary, treatment with somatostatin analogue therapy and treatment with peptide receptor therapy were associated with improved prognosis. Multivariate analysis revealed that age at diagnosis (P=0.014), Ki67 level (P=0.039) and resection of primary (P=0.015) were independent predictors of survival. This is the largest study to our knowledge looking specifically at the prognosis and clinical course of patients with liver metastatic midgut NETs. For the first time, we have shown that Ki67 and resection of primary are independent predictors of survival for this group of patients.


A cell-based system for screening hair growth-promoting agents.
Androgen-inducible transforming growth factor beta (TGF-beta1) derived from dermal papilla cells (DPCs) is a catagen inducer that mediates hair growth suppression in androgenetic alopecia (AGA). In this study, a cell-based assay system was developed to monitor TGF-beta1 promoter activity and then used to evaluate the effects of activated TGF-beta1 promoter in human epidermal keratinocytes (HaCaT). To accomplish this, a pMetLuc-TGF-beta1 promoter plasmid that expresses the luciferase reporter gene in response to TGF-beta1 promoter activity was constructed. Treatment of HaCaT with dihydrotestosterone, which is known to be a primary factor of AGA, resulted in a concentration-dependent increase in TGF-beta1 promoter activity. However, treatment of HaCaT with the TGF-beta1 inhibitor, curcumin, resulted in a concentration-dependant decrease in TGF-beta1 expression. Subsequent use of this assay system to screen TGF-beta1 revealed that HaCaT that were treated with apigenin showed decreased levels of TGF-beta1 expression. In addition, treatment with apigenin also significantly increased the proliferation of both SV40T-DPCs (human DPCs) and HaCaT cells. Furthermore, apigenin stimulated the elongation of hair follicles in a rat vibrissa hair follicle organ culture. Taken together, these findings suggest that apigenin, which is known to have antioxidant, anti-inflammatory, and anti-tumor properties, stimulates hair growth through downregulation of the TGF-beta1 gene. In addition, these results suggest that this assay system could be used to quantitatively measure TGF-beta1 promoter activity in HaCaT, thereby facilitating the screening of agents promoting hair growth.


The reMAP project: A retrospective, 15-year register study on inpatient care for youth with mental disorders.
We aimed to characterize youth hospitalization trends in a psychiatric inpatient unit from a large, public university hospital with a broad catchment area in Milan, Italy. Hospitalization data of patients with an age at admission ≤ 35 were retrospectively retrieved over a time span of 15 years. The sample was comprised of 1982 admissions to a psychiatric ward, aggregated into ICD-10 diagnostic clusters and then analysed. We investigated the epidemiological trends with a focus on age at admission, gender, nationality and hospitalization rates, length of stay and "revolving door" readmissions within a year. Hospitalization rates increased for eating Disorders and decreased for non-affective psychotic disorders; median length of stay generally decreased; hospitalization rates for foreign youth increased, in particular for those diagnosed with non-affective psychotic disorders, personality disorders, and substance-related and addictive disorders. The revolving door phenomenon was also associated with non-affective psychoses and neurodevelopmental disorders, while found to increase for eating disorders. Hospitalization patterns reflect the general increase of foreign youth in the suburban tissue of a large metropolitan area like Milan. However, our data might underestimate the constant growth of mental health problems in foreign youth due to a generally lower access to services. Novel pharmacological treatments and early intervention programs might explain the decrease of hospitalization duration and hospitalization rate for youth with non-affective psychoses. The observed increase in hospitalization for young patients with eating disorders sustains the development of adequate policies tailored towards specialty wards.


The role of temporal parameters in a thalamocortical model of analogy.
How multiple specialized cortical areas in the brain interact with each other to give rise to an integrated behavior is a largely unanswered question. This paper proposes that such an integration can be understood under the framework of analogy and that part of the thalamus and the thalamic reticular nucleus (TRN) may be playing a key role in this respect. The proposed thalamocortical model of analogy heavily depends on a diverse set of temporal parameters including axonal delay and membrane time constant, each of which is critical for the proper functioning of the model. The model requires a specific set of conditions derived from the need of the model to process analogies. Computational results with a network of integrate and fire (IF) neurons suggest that these conditions are indeed necessary, and furthermore, data found in the experimental literature also support these conditions. The model suggests that there is a very good reason for each temporal parameter in the thalamocortical network having a particular value, and that to understand the integrated behavior of the brain, we need to study these parameters simultaneously, not separately.


Multidrug-resistant tuberculosis presenting as a solitary splenic mass in an immunocompetent patient.
Here, we report a case of multidrug-resistant tuberculosis (TB) presenting as a solitary splenic mass in a 60-year-old immunocompetent patient. Splenic TB is unusual and, when present, is usually associated with disseminated disease in immunocompromised patients. A high level of suspicion is required for diagnosis, and, as occurred in our case, it may be an unexpected finding following surgery. Diagnosis was made by polymerase chain reaction, which showed the presence of Mycobacterium tuberculosis DNA. Interestingly, rifampicin- and isoniazid-resistant genes were detected in our analysis. Splenic TB diagnosis and treatment are reviewed.


Functional properties of parietal hand manipulation-related neurons and mirror neurons responding to vision of own hand action.
Parietofrontal pathways play an important role in visually guided motor control. In this pathway, hand manipulation-related neurons in the inferior parietal lobule represent 3-D properties of an object and motor patterns to grasp it. Furthermore, mirror neurons show visual responses that are concerned with the actions of others and motor-related activity during execution of the same grasping action. Because both of these categories of neurons integrate visual and motor signals, these neurons may play a role in motor control based on visual feedback signals. The aim of this study was to investigate whether these neurons in inferior parietal lobule including the anterior intraparietal area and PFG of macaques represent visual images of the monkey's own hand during a self-generated grasping action. We recorded 235 neurons related to hand manipulation tasks. Of these, 54 responded to video clips of the monkey's own hand action, the same as visual feedback during that action or clips of the experimenter's hand action in a lateral view. Of these 54 neurons, 25 responded to video clips of the monkey's own hand, even without an image of the target object. We designated these 25 neurons as "hand-type." Thirty-three of 54 neurons that were defined as mirror neurons showed visual responses to the experimenter's action and motor responses. Thirteen of these mirror neurons were classified as hand-type. These results suggest that activity of hand manipulation-related and mirror neurons in anterior intraparietal/PFG plays a fundamental role in monitoring one's own body state based on visual feedback.


Role of LXCXE motif-dependent interactions in the activity of the retinoblastoma protein.
Cell cycle control by pRb requires the integrity of the pocket domain, which is a region necessary for interactions with a variety of proteins, including E2F and LXCXE-motif containing proteins. Through knowledge of the crystal structure of pRb we have prepared a panel of pRb mutant derivatives in which a cluster of lysine residues that demark the LXCXE peptide binding domain were systematically mutated. One of the mutant derivatives, Rb6A, exhibits significantly reduced LXCXE-dependent interactions with HPV E7, cyclinD1 and HDAC2, but retained LXCXE-independent binding to E2F. Consistent with these results, Rb6A could down-regulate E2F-1-dependent activation of different E2F responsive promoters, but was compromised in Rb-dependent repression. Most importantly, Rb6A retained wild-type growth arrest activity, and colony forming activity similar to wild-type pRb. It is compatible with these results that directly targeting HDAC2 to E2F responsive promoters as an E2F/HDAC hybrid protein failed to effect cell cycle arrest. These results suggest that LXCXE-dependent interactions are not essential for pRb to exert growth arrest.


Tetrathionate-forming thiosulfate dehydrogenase from the acidophilic, chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans.
Thiosulfate dehydrogenase is known to play a significant role in thiosulfate oxidation in the acidophilic, obligately chemolithoautotroph, Acidithiobacillus ferrooxidans. Enzyme activity measured using ferricyanide as the electron acceptor was detected in cell extracts of A. ferrooxidans ATCC 23270 grown on tetrathionate or sulfur, but no activity was detected in ferrous iron-grown cells. The enzyme was enriched 63-fold from cell extracts of tetrathionate-grown cells. Maximum enzyme activity (13.8 U mg(-1)) was observed at pH 2.5 and 70°C. The end product of the enzyme reaction was tetrathionate. The enzyme reduced neither ubiquinone nor horse heart cytochrome c, which serves as an electron acceptor. A major protein with a molecular mass of ∼25 kDa was detected in the partially purified preparation. Heme was not detected in the preparation, according to the results of spectroscopic analysis and heme staining. The open reading frame of AFE_0042 was identified by BLAST by using the N-terminal amino acid sequence of the protein. The gene was found within a region that was previously noted for sulfur metabolism-related gene clustering. The recombinant protein produced in Escherichia coli had a molecular mass of ∼25 kDa and showed thiosulfate dehydrogenase activity, with maximum enzyme activity (6.5 U mg(-1)) observed at pH 2.5 and 50°C.


Effect of indomethacin plus ranitidine in advanced melanoma patients on high-dose interleukin-2.
Preclinical models of advanced melanoma have shown that chronic indomethacin therapy combined with interleukin 2 (IL-2) can eradicate experimental metastases. A phase II trial was done in patients with advanced melanoma. Indomethacin and ranitidine were begun at least one week before IL-2. Of the objective responses in 3 patients, 2 were achieved on ranitidine and indomethacin alone, before start of IL-2. Indomethacin and ranitidine may be responsible for some responses in melanoma patients previously attributed to IL-2.


A new method for calculating femoral anterior cortex point location and its effect on component sizing and placement.
Variation in anterior femoral cortex morphology can cause improper component placement and alignment. When surgical inaccuracies occur, the mechanical properties of the distal femur may be altered, which could result in lower surgical success rates and an increased chance of postoperative complications. The purpose of the study is to come up with a reproducible computational algorithm to simulate what the surgeon does in the operating room. This method could help in surgical preplanning, patient-specific instruments, and implant design. From there, we evaluated (1) the angular difference between reference alignment axes; and (2) whether the location of the anterior cortex point and alignment axes had an effect on implant placement and amount of bone resected in seven implant systems. We analyzed 470 femurs from white and black individuals. Two points were defined using automatic three-dimensional landmarking: sizing point and femoral resection anterior cortex (FRAC) point. Alignment axes including the transepicondylar, posterior condylar, distal anatomical (DAA), and mechanical axes (MAs) were automatically calculated and used along with the resection point to define the anterior reference plane. Two mechanical axes were defined for the purpose of this study: MA-1 is a virtual construct used in navigated surgeries defined as the axis joining the center of the femoral head and the knee center and MA-2 was calculated as the axis joining the center of the femoral head and distal exit point of the DAA. Amounts of anterior, posterior, and distal resected bone were calculated along with the difference in orientation between the alignment axes. The mean angular difference between transepicondylar axis and posterior condylar axis (PCA) was 5.44°±2.99°. All seven implant families showed more total bone resection on both the lateral and medial sides when the implants were aligned using MA-2 and PCA+3 of external rotation (PCA+3) when compared with using MA-1 and PCA+3 (p<0.01). Using MA-2 and PCA+3 as an alignment method reduced the amount of bone resection on both medial and lateral anterior surfaces from 1 to 2 mm. The FRAC point is a key landmark in the placement and sizing of the femoral component. Improper sizing, notching, undercutting, or overstuffing can occur based on selecting the highest or lowest cortex point. Balanced placement, prevention of notching, and anterior and posterior cut balancing were accomplished when using the suggested cortex point.


Delineating the maladaptive pathways of child maltreatment: a mediated moderation analysis of the roles of self-perception and social support.
The current study investigated concurrent and longitudinal mediated and mediated moderation pathways among maltreatment, self-perception (i.e., loneliness and self-esteem), social support, and internalizing and externalizing behavior problems. For both genders, early childhood maltreatment (i.e., ages 0-6) was related directly to internalizing and externalizing behavior problems at age 6, and later maltreatment (i.e., ages 6-8) was directly related to internalizing and externalizing behavior problems at age 8. Results of concurrent mediation and mediated moderation indicated that early maltreatment was significantly related to internalizing and externalizing behavior problems at age 6 indirectly both through age 6 loneliness and self-esteem for boys and through age 6 loneliness for girls. Significant moderation of the pathway from early maltreatment to self-esteem, and for boys, significant mediated moderation to emotional and behavioral problems were found, such that the mediated effect through self-esteem varied across levels of social support, though in an unexpected direction. No significant longitudinal mediation or mediated moderation was found, however, between the age 6 mediators and moderator and internalizing or externalizing problems at age 8. The roles of the hypothesized mediating and moderating mechanisms are discussed, with implications for designing intervention and prevention programs.


Comparison of the distortion of probability information in decision under risk and an equivalent visual task.
Decision makers typically overweight small probabilities and underweight large probabilities. However, there are recent reports that when probability is presented in the form of relative frequencies, this typical pattern reverses. We tested this hypothesis by comparing decision making in two tasks: In one task, probability was stated numerically, and in the other task, it was conveyed through a visual representation. In the visual task, participants chose whether a "stochastic bullet" should be fired at either a large target for a small reward or a small target for a large reward. Participants' knowledge of probability in the visual task was the result of extensive practice firing bullets at targets. In the classical numerical task, participants chose between pairs of lotteries with probabilities and rewards matched to the probabilities and rewards in the visual task. We found that participants' probability-weighting functions were significantly different in the two tasks, but the pattern for the visual task was the typical, not the reversed, pattern.


Coping, stress and negative psychological outcomes in parents of children admitted to a pediatric neurorehabilitation care unit.
Parents' attitudes and psychological adjustment during their child's hospitalization in a pediatric neurorehabilitation care unit are key aspects for the child's adherence to care and the impact of the disease. The aim of this study was to examine the relationship between parenting stress, coping style, and negative psychological outcomes in families of children admitted for the first time to a pediatric neurorehabilitation care unit. This is an observational study. Pediatric neurorehabilitation care unit. One hundred twenty-four parents of children diagnosed with neurodevelopmental or neurological conditions. Parents completed standardized questionnaires assessing parenting stress, coping style, anxiety and depressive symptoms. We found that parents of children with neurodevelopmental conditions showed more emotion-focused coping strategies (P=0.016) and depressive symptoms (P=0.01) compared with parents of children with neurological conditions. Hierarchical regression analyses showed that emotion- and avoidance-oriented coping style and socioeconomic status are crucial factors in the adjustment process of parents of children with neurodevelopmental conditions. By contrast, parenting stress and child difficulties were the most significant predictors of negative psychological outcomes in parents with neurological conditions. This study sought to develop more understanding of the relationship among parenting stress, coping, and anxiety or depressive symptoms in parent of children hospitalized in a pediatric neurorehabilitation care unit. We suggest that examining parents may increase our understanding of the interplay between child and parent functioning in families with children admitted for the first time to a pediatric neurorehabilitation care unit. Identify these predictors might help professionals to develop screening procedures to identify parent at high risk for anxiety or depression, and to conduct early interventions to reduce uncertainty and maladaptive coping strategies that may influences rehabilitation process.


A model of a patient-derived IDH1 mutant anaplastic astrocytoma with alternative lengthening of telomeres.
Mutations in isocitrate dehydrogenase 1 (IDH1) have been found in the vast majority of low grade and progressive infiltrating gliomas and are characterized by the production of 2-hydroxyglutarate from α-ketoglutarate. Recent investigations of malignant gliomas have identified additional genetic and chromosomal abnormalities which cluster with IDH1 mutations into two distinct subgroups. The astrocytic subgroup was found to have frequent mutations in ATRX, TP53 and displays alternative lengthening of telomeres. The second subgroup with oligodendrocytic morphology has frequent mutations in CIC or FUBP1, and is linked to co-deletion of the 1p/19q arms. These mutations reflect the development of two distinct molecular pathways representing the majority of IDH1 mutant gliomas. Unfortunately, due to the scarcity of endogenously derived IDH1 mutant models, there is a lack of accurate models to study mechanism and develop new therapy. Here we report the generation of an endogenous IDH1 anaplastic astrocytoma in vivo model with concurrent mutations in TP53, CDKN2A and ATRX. The model has a similar phenotype and histopathology as the original patient tumor, expresses the IDH1 (R132H) mutant protein and exhibits an alternative lengthening of telomeres phenotype. The JHH-273 model is characteristic of anaplastic astrocytoma and represents a valuable tool for investigating the pathogenesis of this distinct molecular subset of gliomas and for preclinical testing of compounds targeting IDH1 mutations or alternative lengthening of telomeres.


Cellulose Nanofibers from Softwood, Hardwood, and Tunicate: Preparation-Structure-Film Performance Interrelation.
This work reveals the structural variations of cellulose nanofibers (CNF) prepared from different cellulose sources, including softwood (Picea abies), hardwood (Eucalyptus grandis × E. urophylla), and tunicate (Ciona intestinalis), using different preparation processes and their correlations to the formation and performance of the films prepared from the CNF. Here, the CNF are prepared from wood chemical pulps and tunicate isolated cellulose by an identical homogenization treatment subsequent to either an enzymatic hydrolysis or a 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO)-mediated oxidation. They show a large structural diversity in terms of chemical, morphological, and crystalline structure. Among others, the tunicate CNF consist of purer cellulose and have a degree of polymerization higher than that of wood CNF. Introduction of surface charges via the TEMPO-mediated oxidation is found to have significant impacts on the structure, morphology, optical, mechanical, thermal, and hydrophobic properties of the prepared films. For example, the film density is closely related to the charge density of the used CNF, and the tensile stress of the films is correlated to the crystallinity index of the CNF. In turn, the CNF structure is determined by the cellulose sources and the preparation processes. This study provides useful information and knowledge for understanding the importance of the raw material for the quality of CNF for various types of applications.


Cutting edge: cross-presented intracranial antigen primes CD8+ T cells.
The CNS is considered immune privileged due to the blood-brain barrier and the absence of conventional lymphatics. Nonetheless, T cell immune responses specific for CNS Ag have been documented. Where these events are initiated and what cellular mechanisms are involved remain unknown. In this study, we established an experimental mouse model to evaluate the requirements for priming CD8+ T cells following the cross-presentation of intracranial Ag. Surprisingly, we find that even with a damaged blood-brain barrier, Ag presentation occurs in regional lymph nodes and not within the CNS itself. Only once the responding cells have expanded can they traffic to the site of CNS injury. Cross-presentation of intracranial Ag is efficient and the subsequent priming of CD8+ T cells is dependent on CD4+ T cell help and CD40 signaling in host APCs. Our findings have important implications for the initiation of T cell immune responses toward CNS Ags.


Distinguishing between driver and passenger mutations in individual cancer genomes by network enrichment analysis.
In somatic cancer genomes, delineating genuine driver mutations against a background of multiple passenger events is a challenging task. The difficulty of determining function from sequence data and the low frequency of mutations are increasingly hindering the search for novel, less common cancer drivers. The accumulation of extensive amounts of data on somatic point and copy number alterations necessitates the development of systematic methods for driver mutation analysis. We introduce a framework for detecting driver mutations via functional network analysis, which is applied to individual genomes and does not require pooling multiple samples. It probabilistically evaluates 1) functional network links between different mutations in the same genome and 2) links between individual mutations and known cancer pathways. In addition, it can employ correlations of mutation patterns in pairs of genes. The method was used to analyze genomic alterations in two TCGA datasets, one for glioblastoma multiforme and another for ovarian carcinoma, which were generated using different approaches to mutation profiling. The proportions of drivers among the reported de novo point mutations in these cancers were estimated to be 57.8% and 16.8%, respectively. The both sets also included extended chromosomal regions with synchronous duplications or losses of multiple genes. We identified putative copy number driver events within many such segments. Finally, we summarized seemingly disparate mutations and discovered a functional network of collagen modifications in the glioblastoma. In order to select the most efficient network for use with this method, we used a novel, ROC curve-based procedure for benchmarking different network versions by their ability to recover pathway membership. The results of our network-based procedure were in good agreement with published gold standard sets of cancer genes and were shown to complement and expand frequency-based driver analyses. On the other hand, three sequence-based methods applied to the same data yielded poor agreement with each other and with our results. We review the difference in driver proportions discovered by different sequencing approaches and discuss the functional roles of novel driver mutations. The software used in this work and the global network of functional couplings are publicly available at http://research.scilifelab.se/andrej_alexeyenko/downloads.html.


The Shortcomings of COVID-19 Testing in Ecuador: Time to Incentivize Research and Innovation.
The COVID-19 pandemic hit Ecuador severely. The country caught the attention of international media due to its high death toll and overwhelmed healthcare system. The clinical diagnostics system was rapidly overloaded, and the import of PCR tests was delayed. The case of Ecuador illustrates how middle-income countries rely heavily on the importation of biotechnological products for their healthcare systems. The Ecuadorian experience during the COVID-19 pandemic serves as a call for the formation of policies for the development of the biotechnological industry.


Assessment of two-dimensional separative systems using nearest-neighbor distances approach. Part 1: orthogonality aspects.
We propose here a new approach to the evaluation of two-dimensional and, more generally, multidimensional separations based on topological methods. We consider the apex plot as a graph, which could further be treated using a topological tool: the measure of distances between the nearest neighbors (NND). Orthogonality can be thus defined as the quality of peak dispersion in normalized separation space, which is characterized by two factors describing the population of distances between nearest neighbors: the lengths (di(o)) of distances and the degree of similarity of all lengths. Orthogonality grows with the increase of both factors. The NND values were used to calculate a number of new descriptors. They inform about the extent of peak distribution, like the arithmetic mean (A̅(o)) of NNDs, as well as about the homogeneity of peak distribution, like the geometric mean (G̅(o)) and the harmonic mean (H̅(o)). Our new, NND-based approach was compared with another recently published method of orthogonality evaluation: the fractal dimensionality (DF). The comparison shows that the geometric mean (G̅(o)) is the descriptor behaving in the most similar way to dimensionality (DF) and the harmonic mean (H̅(o)) displays superior sensitivity to the shortest, critical distances between peaks. The latter descriptor (H̅(o)) can be considered as sufficient to describe the degree of orthogonality based on NND. The method developed is precise, simple, easy to implement, and possible to use for the description of separations in a true or virtual system of any number of dimensions.


ROCK is involved in vasculogenic mimicry formation in hepatocellular carcinoma cell line.
Ras homolog family member A (RhoA) and Rho-associated coiled coil-containing protein kinases 1 and 2 (ROCK1 and 2) are key regulators of focal adhesion, actomyosin contraction and cell motility. RhoA/ROCK signaling has emerged as an attractive target for the development of new cancer therapeutics. Whether RhoA/ROCK is involved in regulating the formation of tumor cell vasculogenic mimicry (VM) is largely unknown. To confirm this hypothesis, we performed in vitro experiments using hepatocellular carcinoma (HCC) cell lines. Firstly, we demonstrated that HCC cells with higher active RhoA/ROCK expression were prone to form VM channels, as compared with RhoA/ROCK low-expressing cells. Furthermore, Y27632 (a specific inhibitor of ROCK) rather than exoenzyme C3 (a specific inhibitor of RhoA) effectively inhibited the formation of tubular network structures in a dose-dependent manner. To elucidate the possible mechanism of ROCK on VM formation, real-time qPCR, western blot and immunofluorescence were used to detect changes of the key VM-related factors, including VE-cadherin, erythropoietin-producing hepatocellular carcinoma-A2 (EphA2), phosphoinositide 3-kinase (PI3K), matrix metalloproteinase (MMP)14, MMP2, MMP9 and laminin 5γ2-chain (LAMC2), and epithelial-mesenchymal-transition (EMT) markers: E-cadherin and Vimentin. The results showed that all the expression profiles were attenuated by blockage of ROCK. In addition, in vitro cell migration and invasion assays showed that Y27632 inhibited the migration and invasion capacity of HCC cell lines in a dose-dependent manner markedly. These data indicate that ROCK is an important mediator in the formation of tumor cell VM, and suggest that ROCK inhibition may prove useful in the treatment of VM in HCC.


The incidence of diabetes mellitus in Swedish children 1970--1975.
We report a retrospective study of diabetic children, 0--14 years of age, from seven Swedish departments of paediatrics. There were 359 new cases in the years 1970--1975. Notification suggested that there was a mean yearly incidence of 19.6 cases per 100 000 with a year to year variation of 10.0--26.4 per 100 000. Consequently about 330 new cases of childhood diabetes would be expected in Sweden every year. Incidence varied considerably between different geographical areas. The age distribution was bimodal with a main peak at about 12 years and another peak at about 7 years. There was some evidence for clustering of new cases in January and the autumn. The mean prevalence of childhood diabetes in the seven districts was 1.3 per 1 000.


Reappraising the concept of massive transfusion in trauma.
The massive-transfusion concept was introduced to recognize the dilutional complications resulting from large volumes of packed red blood cells (PRBCs). Definitions of massive transfusion vary and lack supporting clinical evidence. Damage-control resuscitation regimens of modern trauma care are targeted to the early correction of acute traumatic coagulopathy. The aim of this study was to identify a clinically relevant definition of trauma massive transfusion based on clinical outcomes. We also examined whether the concept was useful in that early prediction of massive transfusion requirements could allow early activation of blood bank protocols. Datasets on trauma admissions over a 1 or 2-year period were obtained from the trauma registries of five large trauma research networks. A fractional polynomial was used to model the transfusion-associated probability of death. A logistic regression model for the prediction of massive transfusion, defined as 10 or more units of red cell transfusions, was developed. In total, 5,693 patient records were available for analysis. Mortality increased as transfusion requirements increased, but the model indicated no threshold effect. Mortality was 9% in patients who received none to five PRBC units, 22% in patients receiving six to nine PRBC units, and 42% in patients receiving 10 or more units. A logistic model for prediction of massive transfusion was developed and validated at multiple sites but achieved only moderate performance. The area under the receiver operating characteristic curve was 0.81, with specificity of only 50% at a sensitivity of 90% for the prediction of 10 or more PRBC units. Performance varied widely at different trauma centers, with specificity varying from 48% to 91%. No threshold for definition exists at which a massive transfusion specifically results in worse outcomes. Even with a large sample size across multiple trauma datasets, it was not possible to develop a transportable and clinically useful prediction model based on available admission parameters. Massive transfusion as a concept in trauma has limited utility, and emphasis should be placed on identifying patients with massive hemorrhage and acute traumatic coagulopathy.


Noninvasive exploration of cardiac autonomic neuropathy. Four reliable methods for diabetes?
The purpose of this work was to assess relevant information that could be provided by various mathematical analyses of spontaneous blood pressure (BP) and heart rate (HR) variabilities in diabetic cardiovascular neuropathy. There were 10 healthy volunteers and 11 diabetic subjects included in the study. Diabetic patients were selected for nonsymptomatic orthostatic hypotension in an assessment of their cardiovascular autonomic impairment. Cardiac autonomic function was scored according to Ewing's methodology adapted to the use of a Finapres device. The spontaneous beat-to-beat BP and HR variabilities were then analyzed on a 1-h recording in supine subjects. The global variabilities were assessed by standard deviation, fractal dimension, and spectral power. The cardiac baroreflex function was estimated by cross-spectral sequences and Z analyses. In diabetic patients, Ewing's scores ranged from 1 to 4.5, confirming cardiovascular autonomic dysfunction. In these diabetic patients, global indices of variabilities were consistently lower than in healthy subjects. Furthermore, some of them (standard deviation and fractal dimension of HR, spectral power of systolic blood pressure and HR) were significantly correlated with the Ewing's scores. The Z methods and the spectral analysis found that the cardiac baroreflex was less effective in diabetic subjects. However, the baroreflex sensitivity could not be reliably assessed in all the patients. The sequence method pointed out a decreased number of baroreflex sequences in diabetic subjects that was correlated to the Ewing's score. Indices of HR spontaneous beat-to-beat variability are consistently related to the degree of cardiac autonomic dysfunction, according to Ewing's methodology. The Z method and spectral analysis confirmed that the cardiac baroreflex was impaired in diabetic patients. These methods might be clinically relevant for use in detecting incipient neuropathy in diabetic patients.


Discrimination, mental health, and leukocyte telomere length among African American men.
African American men in the US experience disparities across multiple health outcomes. A common mechanism underlying premature declines in health may be accelerated biological aging, as reflected by leukocyte telomere length (LTL). Racial discrimination, a qualitatively unique source of social stress reported by African American men, in tandem with poor mental health, may negatively impact LTL in this population. The current study examined cross-sectional associations between LTL, self-reported racial discrimination, and symptoms of depression and anxiety among 92 African American men 30-50 years of age. LTL was measured in kilobase pairs using quantitative polymerase chain reaction assay. Controlling for sociodemographic factors, greater anxiety symptoms were associated with shorter LTL (b=-0.029, standard error [SE]=0.014; p<0.05). There were no main effects of racial discrimination or depressive symptoms on LTL, but we found evidence for a significant interaction between the two (b=0.011, SE=0.005; p<0.05). Racial discrimination was associated with shorter LTL among those with lower levels of depressive symptoms. Findings from this study highlight the role of social stressors and individual-level psychological factors for physiologic deterioration among African American men. Consistent with research on other populations, greater anxiety may reflect elevated stress associated with shorter LTL. Racial discrimination may represent an additional source of social stress among African American men that has detrimental consequences for cellular aging among those with lower levels of depression.


The effect of regular walks on various health aspects in older people with dementia: protocol of a randomized-controlled trial.
Physical activity has proven to be beneficial for physical functioning, cognition, depression, anxiety, rest-activity rhythm, quality of life (QoL), activities of daily living (ADL) and pain in older people. The aim of this study is to investigate the effect of walking regularly on physical functioning, the progressive cognitive decline, level of depression, anxiety, rest-activity rhythm, QoL, ADL and pain in older people with dementia. This study is a longitudinal randomized controlled, single blind study. Ambulatory older people with dementia, who are regular visitors of daily care or living in a home for the elderly or nursing home in the Netherlands, will be randomly allocated to the experimental or control condition. Participants of the experimental group make supervised walks of 30 minutes a day, 5 days a week, as part of their daily nursing care. Participants of the control group will come together three times a week for tea or other sedentary activities to control for possible positive effects of social interaction. All dependent variables will be assessed at baseline and after 6 weeks, and 3, 6, 9, 12 and 18 months of intervention. The dependent variables include neuropsychological tests to assess cognition, physical tests to determine physical functioning, questionnaires to assess ADL, QoL, level of depression and anxiety, actigraphy to assess rest-activity rhythm and pain scales to determine pain levels. Potential moderating variables at baseline are: socio-demographic characteristics, body mass index, subtype of dementia, apolipoprotein E (ApoE) genotype, medication use and comorbidities. This study evaluates the effect of regular walking as a treatment for older people with dementia. The strength of this study is that 1) it has a longitudinal design with multiple repeated measurements, 2) we assess many different health aspects, 3) the intervention is not performed by research staff, but by nursing staff which enables it to become a routine in usual care. Possible limitations of the study are that 1) only active minded institutions are willing to participate creating a selection bias, 2) the drop-out rate will be high in this population, 3) not all participants will be able to perform/understand all tests. NTR1482.


Drug-induced mRNA signatures are enriched for the minority of genes that are highly heritable.
The blood gene expression signatures are used as biomarkers for immunological and non- immunological diseases. Therefore, it is important to understand the variation in blood gene expression patterns and the factors (heritable/non-heritable) that underlie this variation. In this paper, we study the relationship between drug effects on the one hand, and heritable and non-heritable factors influencing gene expression on the other. Understanding of this relationship can help select appropriate targets for drugs aimed at reverting disease phenotypes to healthy states. In order to estimate heritable and non-heritable effects on gene expression, we use Twin-ACE model on a gene expression dataset MuTHER, measured in blood samples from monozygotic and dizygotic twins. In order to associate gene expression with drug effects, we use CMap database. We show that, even though the expressions of most genes are driven by non-heritable factors, drugs are more likely to influence expression of genes, driven by heritable rather than non-heritable factors. We further study this finding in the context of a gene regulatory network. We investigate the relationship between the drug effects on gene expression and propagation of heritable and non-heritable factors through regulatory networks. We find that the decisive factor in determining whether a gene will be influenced by a drug is the flow of heritable effects supplied to the gene through regulatory network.


Self-assembly of Janus particles under shear.
We investigate the self-assembly of colloidal Janus particles under shear flow by employing hybrid molecular dynamics simulations that explicitly take into account hydrodynamic interactions. Under quiescent conditions, the amphiphilic colloids form spherical micellar aggregates of different sizes, where the solvophobic hemispheres are directed towards the core and the solvophilic caps are exposed to the solvent. When sufficiently strong shear is applied, the micelles disaggregate with a consequent decay of the average cluster size. Nonetheless, we find an intermediate shear rate regime where the balance between rearrangement and dissociation favors the growth of the aggregates. Additionally, our simulations show that clusters composed of either 6 or 13 particles are the most stable towards the shear flow due to their high geometric symmetry. Our findings open up a new range of applications for Janus particles, ranging from biotechnology to sensor systems.


The objective measurement of physical activity and sedentary behaviour in 2-3 year olds and their parents: a cross-sectional feasibility study in the bi-ethnic Born in Bradford cohort.
The reported lower physical activity (PA) levels of British South Asians (SA) are suggested as a key influence in their increased risk of non-communicable diseases compared to their White British peers. Differences in objectively measured PA and sedentary behaviour (SB) between these ethnic groups have been observed during childhood (ages: 8-10 years). However, no information exists on objectively measured PA/SB in younger children, or how early in life differences in these behaviours emerge. Assessing PA/SB in the Born in Bradford (BIB) cohort study provides an opportunity to address such gaps in the literature, but previous studies have found recruiting and retaining SA participants challenging, and the feasibility of using accelerometers with SA children and parents is unknown. This study investigated the feasibility of recruiting and objectively measuring the habitual PA/SB of 2-3 year old SA and White British children and parents from the BIB study. Families were informed about the study during routine BIB assessments. Consenting families were visited at home for anthropometry measurements, interviews, material delivery and collection. Participants (child and parents) were instructed to wear the ActiGraph GT3X+ for 8 days. Descriptive statistics were computed, and ethnic differences tested (Chi-square) for recruitment uptake and compliance. 160 families (30 % SA) provided contact details, and 97 (22 % SA) agreed to enter the study. White British families showed lower refusal and higher intake into the study than SA (p = 0.006). Of 89 children issued with an accelerometer, 34 % complied with the 8-day protocol (significantly less SA; p = 0.015) and 75 % provided enough days (≥ 3) to assess habitual PA/SB (no ethnic differences). Parental rates of compliance with the protocol did not differ between ethnicities. Issues experienced with the protocol and accelerometer use, and successful implementation strategies/procedures are presented. Although greater efforts may be required to recruit SA, those consenting to participate were as likely as White British to provide enough data to assess habitual PA/SB. The issues and successful strategies reported in this feasibility study represent valuable information for planning future studies, and enhance recruitment and compliance with accelerometer protocols in SA and White British toddlers and parents.


Therapeutic effectiveness of percutaneous drainage and factors for performing an interval appendectomy in pediatric appendiceal abscess.
In this study, we studied the therapeutic effectiveness of percutaneous drainage with antibiotics and the need for an interval appendectomy for treating appendiceal abscess in children with a research-oriented dataset released by the Bureau of National Health Insurance in Taiwan through the Collaboration Center for Health Information Application (CCHIA). We identified 1225 patients under 18 years of age who had non-surgical treatment for an appendiceal abscess between 2007 and 2012 in a Taiwan CCHIA dataset. The treatment included percutaneous drainage with antibiotics or antibiotics alone. We also analyzed data of patient's baseline characteristics, outcomes of percutaneous drainage, and indicating factors for performing an interval appendectomy. Totally, 6190 children had an appendiceal abscess, an 1225 patients received non-operative treatment. Of 1225 patients, 150 patients received treatment with percutaneous drainage and antibiotics, 78 had recurrent appendicitis, 185 went on to receive an interval appendectomy, and 10 had postoperative complications after the interval appendectomy. We found that patients treated with percutaneous drainage and antibiotics had a significantly lower rate of recurrent appendicitis (p < 0.05), a significantly smaller chance of receiving an interval appendectomy (p < 0.05), and significantly fewer postoperative complications after the interval appendectomy (p < 0.05) than those without percutaneous drainage treatment. Older children (13 ~ 18 years) patients were found to have a significantly smaller need to receive an interval appendectomy than those who were ≤ 6 years of age (odd ratio (OR) = 2.071, 95 % confidence interval (CI) = 1.34-3.19, p < 0.01), and those who were 7 ~ 12 years old (OR = 1.662, 95 % CI = 1.15-2.41, p < 0.01). In addition, those treated with percutaneous drainage were significantly less indicated to receive an interval appendectomy later (OR = 2.249, 95 % CI = 1.19 ~ 4.26, p < 0.05). In addition, those with recurrent appendicitis had a significantly increased incidence of receiving an interval appendectomy later (OR = 3.231, 95 % CI = 1.95 ~ 5.35, p < 0.001). In this study, we used nationwide data to demonstrate therapeutic effectiveness of percutaneous drainage and antibiotics was more beneficial than only antibiotics in treating patients with an appendiceal abscess. We also found three factors that were significantly associated with receiving an interval appendectomy: recurrent appendicitis, being aged ≤ 13 years, and treatment with antibiotics only.


Inferior Auditory Time Perception in Children With Motor Difficulties.
Accurate time perception is crucial for hearing (speech, music) and action (walking, catching). Motor brain regions are recruited during auditory time perception. Therefore, the hypothesis was tested that children (age 6-7) at risk for developmental coordination disorder (rDCD), a neurodevelopmental disorder involving motor difficulties, would show nonmotor auditory time perception deficits. Psychophysical tasks confirmed that children with rDCD have poorer duration and rhythm perception than typically developing children (N = 47, d = 0.95-1.01). Electroencephalography showed delayed mismatch negativity or P3a event-related potential latency in response to duration or rhythm deviants, reflecting inefficient brain processing (N = 54, d = 0.71-0.95). These findings are among the first to characterize perceptual timing deficits in DCD, suggesting important theoretical and clinical implications.


Mechanistic insights on the copolymerization of polar vinyl monomers with neutral Ni(II) catalysts.
The Ni(II) complexes [(N--O)Ni(H)(PMe(3))] (1) and [(N--O)Ni(CH(2)CH(3))(dmso)] (3) (NwedgeO = kappa(2)-{(2,6-(3,5-(F(3)C)(2)C(6)H(3))(2)C(6)H(3))-N=C(H)-(3,5-I(2)-2-O-C(6)H(2))}) were found to be well-defined model compounds to study the reactivity of polymerization active neutral Ni(II) species toward polar vinyl monomers. Methyl acrylate (MA) insertion into the Ni(II)-hydride bond of 1 was monitored at T >or= -40 degrees C by NMR spectroscopy. 2,1-Insertion yields the functionalized Ni(II) alkyl complex [(N--O)Ni(C(alpha)H(CH(3))C(beta)(O)OCH(3))(PMe(3))] (4). Low-temperature 2D ROESY data indicate a weak Ni(II)...O=C(beta) interaction in 4. This is supported by ab initio calculations at the gradient-corrected DFT (BP86/LACPV*) level of theory. Exposure of 1 to equal amounts of MA and ethylene afforded 4 and the Ni(II) ethyl complex 7 in a 9:1 ratio, which indicates that MA and ethylene effectively compete with each other for coordination and insertion. NMR spectroscopic monitoring revealed that 4 is stable in the absence of residual 1 at low temperatures but is subject to rapid bimolecular elimination of the functionalized alkyl moiety in the presence of free Ni(II) hydride species even at T = -40 degrees C. Isomerization into the 1,2-MA-insertion product was observed at T = 25 degrees C but occurred slowly compared to decomposition, which occurs at 0 degrees C by reaction of 4 with Ni(II) hydride formed by beta-elimination from 4 itself at this temperature. Exposure of the higher Ni(II) alkyl complex 3 to MA in the presence of excess ethylene results in the immediate formation of methyl pentanoate as the ultimate decomposition product. Functionalized Ni(II) alkyl species formed from 2,1-insertion of MA into the metal-carbon bond of higher Ni(II) alkyls are subject to rapid hydrolysis in the presence of trace amounts of water in the reaction medium, which contrasts the stability of nonfunctionalized Ni(II) alkyls toward water. Exposure of 1 to vinyl acetate (VA) affords the kinetic 1,2-insertion product [(NwedgeO)Ni(CH(2)CH(2)OC(gamma')(O)CH(3))(PMe(3))] (5) at temperatures T >or= -10 degrees C, which rearranges into the thermodynamically favored 2,1-insertion product [(N--O)Ni(CH(CH(3))OC(O)CH(3))(PMe(3))] (6). NMR data and ab initio calculations suggest a Ni(II)...O=C(gamma') interaction in 6. 5 decomposes via beta-acetate elimination to yield ethylene and Ni(II) acetate species. Notably, VA does not undergo observable nickel-carbon bond insertion with 3, but reacted with Ni(II) hydride species in equilibrium with 3 to yield 5* which is subject to rapid decomposition via beta-acetate elimination.


Organizational complexity and demographic scale in primary states.
The relationship between organizational complexity and demographic scale is an enduring research problem at the intersection of the natural and social sciences and has far reaching implications for the study of social evolution, particularly the emergence and collapse of complex social organizations such as chiefdoms, states and empires. Anthropological models of social evolution universally assume that population growth plays a critical role in the development of organizational complexity; however, the relationship between organizational complexity and demographic scale has not been formalized and cross-culturally validated. There is a rich yet unsystematized body of diachronic organizational and demographic data describing the evolution of organizational complexity in 10 archaeologically known cases of primary state formation. Using this dataset, this essay proposes and tests a complex network model that describes state societies as discrete, self-similar, hierarchical social networks. The model accurately describes how organizational complexity and population scale in all cases. The complex network architecture of state societies suggests that further advances in our understanding of modern social organization may be found by a deeper investigation of the role of human nature in the evolution of human societies.


Interneuronal exchange and functional integration of synaptobrevin via extracellular vesicles.
Recent studies have investigated the composition and functional effects of extracellular vesicles (EVs) secreted by a variety of cell types. However, the mechanisms underlying the impact of these vesicles on neurotransmission remain unclear. Here, we isolated EVs secreted by rat and mouse hippocampal neurons and found that they contain synaptic-vesicle-associated proteins, in particular the vesicular SNARE (soluble N-ethylmaleimide-sensitive factor [NSF]-attachment protein receptor) synaptobrevin (also called VAMP). Using a combination of electrophysiology and live-fluorescence imaging, we demonstrate that this extracellular pool of synaptobrevins can rapidly integrate into the synaptic vesicle cycle of host neurons via a CD81-dependent process and selectively augment inhibitory neurotransmission as well as specifically rescue neurotransmission in synapses deficient in synaptobrevin. These findings uncover a novel means of interneuronal communication and functional coupling via exchange of vesicular SNAREs.


A unique, highly conserved secretory invertase is differentially expressed by promastigote developmental forms of all species of the human pathogen, Leishmania.
Leishmania are protozoan pathogens of humans that exist as extracellular promastigotes in the gut of their sand fly vectors and as obligate intracellular amastigotes within phagolysosomes of infected macrophages. Between infectious blood meal feeds, sand flies take plant juice meals that contain sucrose and store these sugars in their crop. Such sugars are regurgitated into the sand fly anterior midgut where they impact the developing promastigote parasite population. In this report we showed that promastigotes of all Leishmania species secreted an invertase/sucrase enzyme during their growth in vitro. In contrast, neither L. donovani nor L. mexicana amastigotes possessed any detectable invertase activity. Importantly, no released/secreted invertase activity was detected in culture supernatants from either Trypanosoma brucei or Trypanosoma cruzi. Using HPLC, the L. donovani secretory invertase was isolated and subjected to amino acid sequencing. Subsequently, we used a molecular approach to identify the LdINV and LmexINV genes encoding the ~72 kDa invertases produced by these organisms. Interestingly, we identified high fidelity LdINV-like homologs in the genomes of all Leishmania sp. but none were present in either T. brucei or T. cruzi. Northern blot and RT-PCR analyses showed that these genes were developmentally/differentially expressed in promastigotes but not amastigotes of these parasites. Homologous transfection studies demonstrated that these genes in fact encoded the functional secretory invertases produced by these parasites. Cumulatively, our results suggest that these secretory enzymes play critical roles in the survival/growth/development and transmission of all Leishmania parasites within their sand fly vector hosts.


Characterization of a monocyanide model of FeFe hydrogenases - highlighting the importance of the bridgehead nitrogen for catalysis.
An azadithiolate bridged monocyanide derivative [Fe2(adt)(CO)5(CN)]- of [Fe2(adt)(CO)4(CN)2]2- has been prepared and extensively characterized as a model of the [FeFe]-hydrogenase active site, using a combination of FTIR spectroscopy, electrochemical methods and catalytic assays with chemical reductants. The presence of two basic nitrogen sites opens up multiple protonation pathways, enabling catalytic proton reduction. To our knowledge [Fe2(adt)(CO)5(CN)]- represents the first example of a cyanide containing [FeFe]-hydrogenase active site mimic capable of catalytic H2 formation in aqueous media.


Postnatal Corticosteroids Policy for Very Preterm Infants and Bronchopulmonary Dysplasia.
Postnatal corticosteroids (PNC) are effective for reducing bronchopulmonary dysplasia (BPD) in very preterm neonates but are associated with adverse effects including an increased risk of cerebral palsy. PNC use in Europe is heterogeneous across regions. This study aimed to assess whether European neonatal intensive care units (NICUs) with a low use of PNC or an explicit policy to reduce PNC use had higher risks of mortality or BPD. We included 3,126 infants in 105 NICUs born between 24 + 0 and 29 + 6 weeks' gestational age in 19 regions in 11 countries in the EPICE cohort. First, we identified clusters of NICUs using hierarchical clustering based on PNC use and BPD prevalence and compared case mix and mortality between the clusters. Second, a multilevel analysis was performed to evaluate the association between a restrictive PNC policy and BPD occurrence. There were 3 clusters of NICUs: 52 with low PNC use and a low BPD rate, 37 with low PNC use and a high BPD rate, and 16 with high PNC use and a medium BPD rate. Neonatal mortality did not differ between clusters (p = 0.88). A unit policy of restricted PNC use was not associated with a higher risk of BPD (odds ratio 0.68; 95% confidence interval: 0.45-1.03) after adjustment. Up to 49% of NICUs had low PNC use and low BPD rates, without a difference in mortality. Infants hospitalized in NICUs with a stated policy of low PNC use did not have an increased risk of BPD.