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The potentially detrimental effects of the worldwide deficiency of Omega-3 fatty acids on the COVID-19 pandemic have been underestimated. The Omega-3 Index (O3I), clinical variables, biometric indices, and nutritional information were directly determined for 74 patients with severe COVID-19 and 10 healthy quality-control subjects. The relationships between the OI3 and mechanical ventilation (MV) and death were analyzed. Results: Patients with COVID-19 exhibited low O3I (mean: 4.15%; range: 3.06-6.14%)-consistent with insufficient fish and Omega-3 supplement consumption, and markedly lower than the healthy control subjects (mean: 7.84%; range: 4.65-10.71%). Inverse associations were observed between O3I and MV (OR = 0.459; C.I.: 0.211-0.997) and death (OR = 0.28; C.I.: 0.08-0.985) in severe COVID-19, even after adjusting for sex, age, and well-known risk factors. Conclusion: We present preliminary evidence to support the hypothesis that the risk of severe COVID-19 can be stratified by the O3I quartile. Further investigations are needed to assess the value of the O3I as a blood marker for COVID-19.

Exotic fruits and their co-products may be valuable sources of antioxidant dietary fibres (DF) which are useful for food industry and human health. In this study, we aimed to characterize DF obtained from guavira fruit pomace and investigate its antioxidant potential employing TEAC assay as well as a cell model. The DF were chemically characterized as containing arabinan, highly-methoxylated homogalacturonan and arabinogalactan. The DF-containing fraction (CPW) presented ABTS free radical scavenger activity. MTT and DCFH-DA assay were performed to assess, respectively, changes in cell viability and the potential intracellular antioxidant activity against H 2 O 2 -induced oxidative stress in murine NIH 3T3 fibroblast. CPW exhibited no effects on cell viability, moreover, when administered 48 h prior the induction of H 2 O 2 toxic effects, it protected the cells, significantly increasing the cell viability compared to control. This protection may be related to the observed reduction of reactive oxygen species levels. Thus, the pre-treatment of cells with guavira DF for 48 h remarkably induced a cytoprotection against pro-oxidant conditions, and may be a valuable functional compound recovered from an unexploited agroindutrial waste., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: The mother-child relationship quality (MCRQ) may represent a transgenerational transmission mechanism of mental health problems. In this context, we examine the mother's recalled parental rearing behavior, actual attributions of her child's behavior, and her current psychopathology., Method: A clinical sample of mother-child dyads was assessed with the Questionnaire for the Assessment of Recalled Parental Rearing Behavior, the Parent Cognition Scale, and the Symptom Checklist at the Child Psychiatric Family Day Hospital for preschool children in Münster, Germany, at admission. MCRQ was assessed with the Parent-Infant Relationship Global Assessment Scale, a structured interview with the child (Strukturiertes Interview zur Erfassung der Kind-Eltern-Interaktion), and the Multiperspective Parent-Child Relationship Questionnaire., Results: Multiple regression analyses showed no direct association between the mother's recalled parental rearing behavior and any measure of the MCRQ. However, maternal dysfunctional attributions about her child's behavior and her actual psychopathology showed the expected negative associations with the multiperspective measures of MCRQ. The relationship quality assessments did not correlate significantly with each other., Conclusion: The divergent measures of MCRQ, which seem to assess different aspects, are a barrier to investigate the association between the mother's recalled parental rearing behavior and MCRQ. However, low MCRQ is associated with increased maternal psychopathology and maternal dysfunctional attributions on child behavior., (© 2020 The Authors. Infant Mental Health Journal published by Wiley Periodicals, Inc. on behalf of Michigan Association for Infant Mental Health.)

Objective: To report the experience with prenatal repair of open spina bifida (OSB) from 2 centers in Chile., Methods: Women with a second-trimester fetus with OSB were offered intrauterine neurosurgical repair following the protocol from the Management of Myelomeningocele Study (MOMS) trial. Pediatric follow-up with infants reaching 12 and 30 months of life was also reviewed., Results: Fifty-eight fetuses with OSB underwent intrauterine repair at an average (±SD) gestational age of 24.8 ± 0.9 weeks. There were 3 (5.1%) intrauterine deaths. The average gestational age at delivery of the remaining 55 cases was 33.3 ± 3.6 weeks, and the average birth weight was 2,172 ± 751 g. Delivery before 30 weeks occurred in 11 cases (20.0%). Two (3.6%) neonatal deaths (<28 days) occurred. At 12 months, a ventriculoperitoneal shunt or an endoscopic third ventriculostomy was required in 25% of the cases. At 30 months, 72.4% of the infants were able to walk., Discussion: Prenatal neurosurgical repair of OSB is a complex and challenging intervention. Major complications include perinatal death and severe prematurity. No major maternal complications occurred in our series. A reduction in the need for cerebrospinal fluid diversion and an improved ability to walk seem to be the greatest long-term advantages of this procedure., (© 2020 S. Karger AG, Basel.)

The Emotional Competence of Mothers and the Clinical Quality of Mother-Child Relationship in a Preschool Psychiatric Population The quality of mother-child interaction and relationship is an essential risk factor for the development of mental disorders at preschool age. We examine maternal emotional competence and maternal level of psychopathology as predictors for the quality of mother-child relationship. At the beginning of their treatment a clinical sample of mother-child-dyads at the Child Psychiatric Family Day Hospital in Münster was assessed concerning the emotional competence of mothers (EKF), the maternal psychopathology (SCL) and the mother-child relationship quality, the latter with three different instruments reflecting the mother's, the child's and the therapist's perspective (PIR-GAS, SKEI, M-PCR). The mothers showed decreased results in the EKF scales emotional regulation, recognition of emotions and in the EKF-overall-score and presented a higher level of maternal psychopathology compared to the normal population. There is a direct relation between emotional regulation and the M-PCR scales affective bond and functional-conflict. Besides the direct influence of the maternal psychopathology, the expected moderator effect of emotional expressiveness and the M-PCR scale functional-conflict, reported by mothers, was apparent. Certain aspects of the emotional competence of mothers and their psychopathology show an influence on mother-child relationship quality. The method and the perspective of assessment are crucial to the results.

Fluctuations in parenting behaviour are thought to be important for the development of child psychopathology. This study focusses on fluctuations in the parenting behaviour of mothers with 3-6-year-old children with a clinical diagnosis according to the International Statistical Classification of Diseases and Related Health Problems 10th Revision (ICD-10) (N = 39) and compared them with a control group of mothers with children without a clinical diagnosis (N = 41). In a laboratory setting, we compared the quality of mother-child interactions between both groups using three increasingly challenging co-operation tasks. At first, the mother and child interacted via a free play task. They then co-operated within a constructional play task and finally within a challenging problem-solving task. We analysed the mothers' parenting behaviour using the Laboratory Parenting Assessment Battery (LAB-PAB) and children's problem behaviours by means of their mothers' rating using the Child Behavior Checklist 1 ½-5 (CBCL). The results corroborated our hypotheses. Mothers of the group of children with clinical diagnoses had a lower parenting quality and higher fluctuations in parenting behaviour across situations compared with the non-clinical group. Further analysis revealed that specific fluctuations in maternal involvement and hostility uniquely predicted child psychopathology, measured with the CBCL, showing incremental validity of fluctuations in maternal involvement, when controlling for parenting quality and maternal difficulties in emotion regulation, measured with the Difficulties in Emotional Regulation scale. The results are discussed in terms of their implications for clinical interventions, as well as theoretical implications and future research.

A new source of pectin with a cytotoxic effect on glioblastoma cells is presented. A homogeneous GWP-FP-S fraction (M w of 29,170 g mol -1 ) was obtained by fractionating the crude pectin extract (GW) from Campomanesia xanthocarpa pulp. According to the monosaccharide composition, the GWP-FP-S was composed of galacturonic acid (58.8%), arabinose (28.5%), galactose (11.3%) and rhamnose (1.1%), comprising 57.7% of homogalacturonans (HG) and 42.0% of type I rhamnogalacturonans (RG-I). These structures were characterized by chromatographic and spectroscopic methods; GW and GWP-FP-S fractions were evaluated by MTT and crystal violet assays for their cytotoxic effects. Both fractions induced cytotoxicity (15.55-37.65%) with concomitant increase in the cellular ROS levels in human glioblastoma cells at 25-400 μg mL -1 , after 48 h of treatment, whereas no cytotoxicity was observed for normal NIH 3T3 cells. This is the first report of in vitro bioactivity and the first investigation of the antitumor potential of gabiroba pectins., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Chagas disease is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi. It is a significant public health problem, affecting millions of people worldwide. And although it was described 110 years ago, only two old nitroheterocyclic drugs, benznidazole and nifurtimox, are currently available for the treatment of Chagas disease and both have several limitations. Besides the clear unmet medical need, many challenges preclude the development of new treatments, some of them related to a lack of understanding of the pathophysiology of the disease and parasite-host interactions. New knowledge and tools are becoming available, but the number of new chemical entities progressing through the preclinical pipeline is inadequate. Therefore, it is still uncertain whether safe, effective and accessible new drugs will be available in the near future. The Chagas disease research community must commit to even greater collaboration to ensure that patients eventually benefit from better treatments., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Most longitudinal or follow-up mental health studies describe developmental pathways using dimensional measures of psychopathology, but seldom using pathways described by clinical disorders. Objective: We aim to describe diagnostic pathways by homotypic (within the disorder continuity) and heterotypic development (between the disorder continuity), with maternal psychopathology as moderator for both trajectories. Methods: Clinically referred children (0-7 years; N = 83) were assessed at preschool age and at primary-school age through a clinical interview. We built a disorder cluster of emotional disorders (ED; F32, F40, F42, F43, F93.0, F93.1, F93.2, F93.8, F95), behavioral disorders (BD; F68.8, F90, F91, F91.3, F93.3, F93.9, F94), and specific early onset disorders (SEO; F50, F51, F70, F98.0, F98.1, F98.2, F98.8, F98.9). We describe the prevalence, comorbidity, and clinical trajectories of various types of homotypic and heterotypic development. Results: We observed a high rate of comorbidity throughout the study (62.6% at admission and 67.5% at follow-up) and in general, a high continuity of mental health problems from preschool to primary-school age children (69.9% of the sample showed continuity), with 50.6% of the sample showing homotypic and 44.6% showing heterotypic development. Hierarchical multiple regression analyses suggest that heterotypic development may be influenced by maternal psychopathology. Conclusion: Currently, evidence-based mental health guidelines for preschool populations are designed and evaluated assuming a homotypic development. However, our findings indicate that treatment interventions and outcome measures should also be designed and evaluated for heterotypic development especially in case of increased maternal psychopathology.

Introduction: Chagas disease (CD) is one of the most neglected public health problems in the Americas, where <1% of the estimated 6 million people with the infection have been diagnosed and treated. The goal of treatment is to eliminate the parasite, decrease the probability of cardiomyopathy and other complications during the chronic stage of infection, and interrupt the cycle of disease transmission by preventing congenital infection. Currently, only benznidazole (BZN) and nifurtimox are recognized by the World Health Organization as effective drugs for treatment of CD. In this paper, we provide an overview of the clinical pharmacology of BZN. Areas covered: This review covers the historical background, chemistry, mechanism of action, pharmacokinetics, preclinical research, resistance, clinical research, toxicology, adverse effects, and current regulatory status of BZN. Expert commentary: Ongoing investigations aim to optimize BZN therapy by adjusting the current standard regimen or by combining BZN with new chemical entities. These studies are assessing alternatives that improve safety while maintaining or increasing the efficacy of BZN. Timely diagnosis and antitrypanosomal treatment are critical components of programs to eliminate CD as a public health problem, and can dramatically reduce the heavy burden of morbidity and mortality caused by the disease.

Background: The advent of less radical surgical approaches has generated concern about leaving locoregional lymph node metastases (LNM) unresected that could lead to adverse outcome. We examined the prognostic role of clinicopathological factors for ypN-positivity in patients with ypT0-2 rectal carcinoma treated within the CAO/ARO/AIO-94 and CAO/ARO/AIO-04 randomized phase 3 trials., Methods: The correlation of clinicopathological factors with ypN-status (ypN0 vs ypN1/2) was examined in n = 776 patients with ypT0-2 rectal carcinoma after preoperative CRT and total mesorectal excision surgery using Pearson's Chi-squared test for categorical variables and Kruskal-Wallis' test for continuous variables. Multivariable analysis was performed using binary logistic regression to identify independent prognosticators for ypN-positivity., Results: Residual LNM (ypN+) were found in 6%, 20.8% and 21.4% of patients with ypT0, ypT1 and ypT2 carcinomas, respectively. Independent prognosticators for LNM were advanced ypT category (p = 0.002) and lymphatic invasion (p = 0.020). In a separate multivariable analysis performed upon exclusion of ypT-category due to multicollinearity with residual tumor diameter (RTD), lymphatic invasion (p = 0.015) and RTD ≥10 mm (p = 0.005) demonstrated strong correlation with LNM., Conclusion: Advanced ypT-stage, lymphatic invasion and RTD ≥10 mm were prognostic factors for LNM in patients ypT0-2 rectal carcinoma treated with CRT and surgery within both phase 3 trials. The high incidence of LNM in the ypT1-2 group needs to be taken into consideration in the context of oncological safety and indicate that LE should be advocated with great caution in this patient subgroup. The prognostic pathological factor identified here could help guide decision of LE vs TME after standard CRT., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The Multiperspective Parent-Child Relationship Questionnaire (M-PCR) Parent-child relationship quality may considerably impact child mental health. Yet many instruments apply effortful behavioral observation methods. In this paper we aim to develop a multiperspective questionnaire of parent-child relationships. Several experts in the field of parent-child interaction treatment generated 18 items to assess aspects of the parent-child relationship. Mothers, therapists and assistant therapists rated 129 mother-child dyads that were treated at the Child Psychiatric Family Day Hospital for preschool children in Münster, Germany at the beginning and again at the end of treatment. For validity purposes, a global measure of relationship quality (PIRGAS) and measures of maternal psychopathology (SCL-90-R) and child behavior problems (CBCL/1.5-5; C-TRF/1.5-5) completed the assessment.Six principal component analyses and subsequent oblique rotation revealed a stable patternof two interpretable factors labeled as "Affective Bond" and "Functional-Conflict" with highinternal consistency and retest-reliability. Validity of the MEK scales was supported by significant correlations with the global rating of relationship quality PIRGAS by experts (therapistsand assistant therapists) and measures of maternal and child psychopathology, particularlyexternalizing problems, and therapy outcome. The MEK appears as a valid, reliable and user-friendly instrument for assessing the quality of parent-child relationship in clinical contextsfrom different perspectives.

In addition to its function as an inhibitor of histone acetyltransferases, Nir (Noc2l) binds to p53 and TAp63 to regulate their activity. Here, we show that epidermis-specific ablation of Nir impairs epidermal stratification and barrier function, resulting in perinatal lethality. Nir-deficient epidermis lacks appendages and remains single layered during embryogenesis. Cell proliferation is inhibited, whereas apoptosis and p53 acetylation are increased, indicating that Nir is controlling cell proliferation by limiting p53 acetylation. Transcriptome analysis revealed that Nir regulates the expression of essential factors in epidermis development, such as keratins, integrins and laminins. Furthermore, Nir binds to and controls the expression of p63 and limits H3K18ac at the p63 promoter. Corroborating the stratification defects, asymmetric cell divisions were virtually absent in Nir-deficient mice, suggesting that Nir is required for correct mitotic spindle orientation. In summary, our data define Nir as a key regulator of skin development., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

Malpighia emarginata is a tropical fruit plant, found naturally in the Caribbean islands and South America that produces an edible fruit known as acerola or Barbados Cherry. Its polysaccharides were obtained by aqueous extraction, subjected to a freezing and thawing process and ultrafiltration. A homogeneous fraction (ACWS-01E) was analyzed by sugar composition, HPSEC, methylation and NMR spectroscopy analyses. The results showed an arabinan-rich pectic polysaccharide, with 6.1×10 4 g/mol and formed mainly by a high methyl esterified (DM=86%) homogalacturonan and branched arabinan. This latter is anchored in type I rhamnogalacturonan regions. The main chain of arabinan consisted of (1→5)-linked α-Araf, branched only at O-3. The potential ACWS-01E intracellular antioxidant activity against H 2 O 2 -induced oxidative stress in murine fibroblast cell line (3T3) was determined by DCFH-DA assay. The treatment with ACWS-01E significantly reduced H 2 O 2 -induced cytotoxic effect and the levels of reactive oxygen species (ROS). These findings suggested that ACWS-01E protected and improved NIH 3T3 cell viability from H 2 O 2 -induced toxicity by decreasing intracellular levels of ROS., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The indications for cochlear implantation (CI) have expanded over the last few years. There is evidence that some adult patients with pre- or perilingual onset of deafness may gain from implantation. Similarly, CI in patients with single-sided deafness may offer significant benefits in terms of quality of life and social as well as academic development. In this setting, directional hearing may be restored and speech comprehension, especially in noise, may be optimized. In patients with intractable tinnitus and profound hearing loss, CI not only improves speech perception, but also helps to reduce the tinnitus in the deaf ear., (© 2018 S. Karger AG, Basel.)

Clinical Validation of the Caregiver-Child Socioemotional and Relationship Rating Scale (SIRS) for Child Behavior in a Preschool-Age Sample The description of child behavior in mother-child-interaction is important in early detection and treatment of psychiatric disorders in preschool children. The Caregiver-Child Socioemotional and Relationship Rating Scale (SIRS) may serve this diagnostic purpose. We aim to examine interrater-reliability of SIRS and concurrent, convergent, and discriminant validity to maternal behavior by Play-PAB, and a measure of mother-child-relationsship by Parent-Infant-Global-Assessment-Scale (PIRGAS). Five raters assessed 47 ten-minute video sequences of parent-child-interaction recorded at the Family Day Hospital for Preschool Children with SIRS, Play-PAB, and PIRGAS. We report psychometric properties of SIRS, and present the association with Play-PAB and PIRGAS. SIRS shows a satisfying interrater-reliability for all items. Positive child behavior e. g. the SIRS' "child responsiveness" shows negative correlation to Play-PAB-scales' parental "hostility" and "intrusiveness", but independence of parental "involvement", "positive emotionality", and "discipline". Child and parental behavior show expected associations with the global relationship measure PIRGAS. The assessment of child behavior in parent-child-interaction with SIRS can be quickly learned and reliably applied without extensive training. SIRS shows meaningful relations to parental behavior and a clinical global measure of the caregiver-child-relationship. We recommend SIRS for clinical diagnostics to describe child behavior in mother-child-interaction.

Introduction: In Internet forums and other social media many reports regarding chronic headaches after cochlear implantation can be found. Although quite rare, there are also some reports in the literature. However, little is known regarding the true prevalence of headaches in persons who have undergone cochlear implant surgery., Objectives: The primary aim of this study was to investigate the 1-year prevalence of headache in patients having received a cochlear implantation ("cochlear implant group") in comparison with patients having undergone middle ear surgery ("surgery group") and persons with no history of head and neck surgery ("non-ear-nose-throat [ENT] group")., Methods: Cross-sectional, monocentric study using a validated headache questionnaire., Results: Three hundred persons were asked to participate. Two hundred thirty four valid questionnaires were returned. The participants' median age was 62 years, of whom 45% were women. The prevalence of headache was 31% (95%-confidence interval [CI]: [21; 42]) in the cochlear implant group and 46% (95%-CI: [35; 57]) in the surgery group with no significant difference between these two subgroups (p = 0.071). In the non-ENT group the prevalence of headache was significantly higher than in the other two subgroups (64%, 95%-CI: [52; 74])., Discussion: The prevalence of headache is not higher in cochlear implant patients in comparison to middle ear surgery patients, other, non-ENT patients and the general German or European population., Conclusion: Cochlear implantation does not seem to be associated with an increased risk for developing headache.

Previous work indicated that lysine-specific demethylase 1 (Lsd1) can positively regulate the oxidative and thermogenic capacities of white and beige adipocytes. Here we investigate the role of Lsd1 in brown adipose tissue (BAT) and find that BAT-selective Lsd1 ablation induces a shift from oxidative to glycolytic metabolism. This shift is associated with downregulation of BAT-specific and upregulation of white adipose tissue (WAT)-selective gene expression. This results in the accumulation of di- and triacylglycerides and culminates in a profound whitening of BAT in aged Lsd1-deficient mice. Further studies show that Lsd1 maintains BAT properties via a dual role. It activates BAT-selective gene expression in concert with the transcription factor Nrf1 and represses WAT-selective genes through recruitment of the CoREST complex. In conclusion, our data uncover Lsd1 as a key regulator of gene expression and metabolic function in BAT., (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)

Streptavidin is a homotetrameric protein binding the vitamin biotin and peptide analogues with an extremely high affinity, which leads to a large variety of applications. The biotin-auxotrophic yeast Pichia pastoris has recently been identified as a suitable host for the expression of the streptavidin gene, allowing both high product concentrations and productivities. However, so far only methanol-based expression systems have been applied, bringing about increased oxygen demand, strong heat evolution and high requirements for process safety, causing increased cost. Moreover, common methanol-based processes lead to large proportions of biotin-blocked binding sites of streptavidin due to biotin-supplemented media. Targeting these problems, this paper provides strategies for the methanol-free production of highly bioactive core streptavidin by P. pastoris under control of the constitutive GAP promoter. Complex were superior to synthetic production media regarding the proportion of biotin-blocked streptavidin. The optimized, easily scalable fed-batch process led to a tetrameric product concentration of up to 4.16 ± 0.11 µM of biotin-free streptavidin and a productivity of 57.8 nM h(-1) based on constant glucose feeding and a successive shift of temperature and pH throughout the cultivation, surpassing the concentration in un-optimized conditions by a factor of 3.4. Parameter estimation indicates that the optimized conditions caused a strongly increased accumulation of product at diminishing specific growth rates (μ ≈ D < 0.01 h(-1) ), supporting the strategy of feeding. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:855-864, 2016., (© 2016 American Institute of Chemical Engineers.)

Streptavidin - a protein secreted by the filamentous bacterium Streptomyces avidinii - is applied in a variety of methods, leading to numerous studies on its heterologous production. Development and characterization of a novel expression system for streptavidin genes by Hansenula polymorpha is described utilizing different target gene variants along with the two methanol-inducible promoters PMOX and PFMD. Extracellular product concentrations were higher for cultivation at 30 instead of 37°C. The best performing strain carrying the full-length streptavidin gene under control of PFMD was characterized in the bioreactor applying a synthetic medium and oxygen-controlled feeding of glucose. Derepression resulted in an extracellular concentration of 1.31±0.07μM of tetrameric streptavidin after 48h (27.3nMh(-1)). Feeding of glycerol improved biomass formation, but lowered the product concentration. By combining derepression and methanol induction the final extracellular streptavidin concentration increased to 11.42±0.22μM (approx. 751mgL(-1)), yielding a productivity of 52.5nMh(-1). Despite supplementing biotin the proportion of biotin-blocked binding sites in the supernatant dropped from 54.4±5.0 % after 18h to 17.2±6.5 % towards the end of glucose feeding to a final value of 1.1±3.8 %, indicating a highly bioactive product. Thus, H. polymorpha proved to be a suitable host for the production of streptavidin., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A regio- and stereoselective reverse prenylation of indole and tryptophan derivatives is presented. All four possible stereoisomers are accessible through this iridium-catalyzed reaction. The stereoselectivity is controlled by a chiral phosphoramidite ligand in combination with an achiral borane additive and can be switched by changing the nature of the borane. One enantiomer of the ligand is thus sufficient to prepare all possible isomers. The synthetic potential of this method was demonstrated by a short total synthesis of amauromine and its two natural diastereomers., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Due to its various applications the protein streptavidin is a highly interesting target for heterologous production. This study focuses on different Escherichia coli-based constructs targeting a high-level expression and secretion of streptavidin to the medium. The effect of various promoters, variants of the target gene, leader sequences and host strains on expression and secretion into the culture broth was analyzed. Constitutive production of full-length streptavidin fused with the leader sequence of the bglA gene from Bacillus amyloliquefaciens by the periplasmic 'leaky mutant' E. coli JW1667-5 (Δlpp-752:kan) at 30°C generated the highest yield of the conditions tested, surpassing the extracellular concentration of a conventional T7-based expression system. Supplementation of the medium by the non-ionic surfactants Triton(®) X-100 and X-45 led to an improved secretion of the protein to the culture supernatant. Tetrameric concentrations of streptavidin of 2790±166nM were reached in shake flasks at a productivity of 49.6nMh(-1). Optimization of conditions led to a successful transfer to the bioreactor, yielding a maximal concentration of 2608±169nM and a productivity of 65.2nMh(-1) in fed-batch operation. The proportion of biotin-blocked binding sites of 8.3±4.3% indicated a highly bioactive product., (Copyright © 2016 Elsevier B.V. All rights reserved.)

There has been an ongoing interest in the analysis and comparison of the efficiency of health care systems using nonparametric and parametric applications. The objective of this study was to review the current state of the literature and to synthesize the findings on health system efficiency in OECD countries. We systematically searched five electronic databases through August 2014 and identified 22 studies that analyzed the efficiency of health care production at the country level. We summarized these studies with view on their sample, methods, and utilized variables. We developed and applied a checklist of 14 items to assess the quality of the reviewed studies along four dimensions: reporting, external validity, bias, and power. Moreover, to examine the internal validity of findings we meta-analyzed the efficiency estimates reported in 35 models from ten studies. The qualitative synthesis of the literature indicated large differences in study designs and methods. The meta-analysis revealed low correlations between country rankings suggesting a lack of internal validity of the efficiency estimates. In conclusion, methodological problems of existing cross-country comparisons of the efficiency of health care systems draw into question the ability of these comparisons to provide meaningful guidance to policy-makers., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Hydrochlorothiazide is a thiazide diuretic widely used in clinics to treat arterial hypertension. It is a class IV drug according to the Biopharmaceutical Classification System, that is, it presents low solubility and low permeability and, consequently, low absorption in the gastrointestinal tract. As a strategy to improve stability and biopharmaceutical properties of hydrochlorothiazide, the use of cyclodextrins to produce inclusion complexes, applying different methods, was investigated. In the phase solubility studies, β-cyclodextrin was identified as the cyclodextrin which provided the most promising results in terms of the solubilization of the drug. The thermal analysis verified the interaction between hydrochlorothiazide and β-cyclodextrin, indicating the formation of inclusion complexes, and the thermal stability varied according to the preparation technique. The physicochemical characterization showed that in the inclusion complexes obtained by co-evaporation, kneading followed by freeze-drying and kneading followed by spray-drying the hydrochlorothiazide complexation mostly occurred with different degrees of amorphization and the drug solubility was improved. These three inclusion complexes presented better in vitro characteristics and the inclusion complex obtained by kneading followed by freeze-drying increased the in vivo diuretic activity of the drug accompanied by significant effects on natriuresis, kaliuresis and chloriuresis. The inclusion complex formation was effective in improving the biopharmaceutical properties of hydrochlorothiazide and protecting the drug from hydrolysis. This paper describes an important alternative approach to the development of liquid pharmaceutical formulations to pediatric administration, a real need of the current pharmaceutical market., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In this study the use of Euglena gracilis biomass for α-tocopherol, paramylon and biogas production in a value-added chain was investigated. Therefore, we analyzed the dry cell weight and product concentrations at different growth phases during heterotrophic, photoheterotrophic and photoautotrophic cultivation in a low-cost minimal medium. Furthermore, the specific biogas yields for differently derived biomass with and without product recovery were investigated. We demonstrate that growth phase and cultivation mode not only have a significant impact on product formation, but also influence the yield of biogas obtained from anaerobic digestion of Euglena gracilis biomass. The maximum dry cell weight concentration ranged from 12.3±0.14gL(-1) for heterotrophically to 3.4±0.02gL(-1) for photoautotrophically grown Euglena gracilis cells. The heterotrophically grown biomass accumulated product concentrations of 5.3±0.12mgL(-1) of α-tocopherol and 9.3±0.1gL(-1) of paramylon or 805±10.9mL of biogasgvs(-1) (per gram volatile solids). The results for photoautotrophically grown cells were 8.6±0.22mgL(-1) of α-tocopherol and 0.78±0.01gL(-1) of paramylon or 648±7.2mL of biogasgvs(-1). For an energy-saving downstream procedure the extracting agent methanol does not have to be removed strictly. Samples with residual methanol showed a significantly increased biogas yield, because the solvent can be used as an additional substrate for methane production by archaebacteria., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Nonsmall cell lung cancer (NSCLC) represents an important cause of mortality worldwide due to its aggressiveness and growing resistance to currently available therapy. Cucurbitacins have emerged as novel potential anticancer agents showing strong antiproliferative effects and can be promising candidates for combined treatments with clinically used anticancer agents. This study investigates the synergistic antiproliferative effects of a new semisynthetic derivative of cucurbitacin B (DACE) with three chemotherapy drugs: cisplatin (CIS), irinotecan (IRI), and paclitaxel (PAC) on A549 cells. The most effective combinations were selected for studies of the mechanism of action. Using an in silico tool, DACE seems to act by a different mechanism of action when compared with that of different classes of drugs already used in clinical settings. DACE also showed potent synergic effects with drugs, and the most potent combinations induced G2/M cell cycle arrest by modulating survivin and p53 expression, disruption of F-actin cytoskeleton, and cell death by apoptosis. These treatments completely inhibited the clonogenic potential and did not reduce the proliferation of nontumoral lung cells (MRC-5). DACE also showed relevant antimigratory and anti-invasive effects, and combined treatments modulated cell migration signaling pathways evolved with metastasis progression. The effects of DACE associated with drugs was potentiated by the oxidant agent l-buthionine-sulfoximine (BSO), and attenuated by N-acetilcysteine (NAC), an antioxidant agent. The antiproliferative effects induced by combined treatments were attenuated by a pan-caspase inhibitor, indicating that the effects of these treatments are dependent on caspase activity. Our data highlight the therapeutic potential of DACE used in combination with known chemotherapy drugs and offer important insights for the development of more effective and selective therapies against lung cancer.

Background: Objective measures of physical activity are currently not considered in clinical guidelines for the assessment of hyperactivity in the context of Attention-Deficit/Hyperactivity Disorder (ADHD) due to low and inconsistent associations between clinical ratings, missing age-related norm data and high technical requirements., Methods: This pilot study introduces a new objective measure for physical activity using compressed webcam video footage, which should be less affected by age-related variables. A pre-test established a preliminary standard procedure for testing a clinical sample of 39 children aged 6-16 years (21 with a clinical ADHD diagnosis, 18 without). Subjects were filmed for 6 min while solving a standardized cognitive performance task. Our webcam video-based video-activity score was compared with respect to two independent video-based movement ratings by students, ratings of Inattentiveness, Hyperactivity and Impulsivity by clinicians (DCL-ADHS) giving a clinical diagnosis of ADHD and parents (FBB-ADHD) and physical features (age, weight, height, BMI) using mean scores, correlations and multiple regression., Results: Our video-activity score showed a high agreement (r = 0.81) with video-based movement ratings, but also considerable associations with age-related physical attributes. After controlling for age-related confounders, the video-activity score showed not the expected association with clinicians' or parents' hyperactivity ratings., Conclusions: Our preliminary conclusion is that our video-activity score assesses physical activity but not specific information related to hyperactivity. The general problem of defining and assessing hyperactivity with objective criteria remains.

Acyclovir, an analog of 2'-deoxyguanosine, is one of the most important drugs in the current approved antiviral treatment. However, it's biopharmaceutical properties, contribute to acyclovir's poor oral bioavailability, which restricts the clinical use of the drug. In this view, the aim of this work was to improve the dissolution rate and intestinal permeability of acyclovir through the development of ball milling solid dispersions with the hydrophilic carriers Pluronic F68®, hydroxypropylmethyl cellulose K100M® and chitosan. Solid dispersions were obtained and completely characterized through different solid state techniques. The solid state data demonstrated a decrease in the crystallinity (amorphous phase and defects) and the presence of hydrogen bonds for SD HPMC and SD CTS. The enhancement of dissolution rates was observed for all SDs developed. In addition, no detrimental effects over the in vitro antiviral activity were detected. The solid dispersions with Pluronic F68® significantly improved the intestinal permeability of acyclovir across Caco-2 cells. In summary, the SDs developed in this study could be considered as potential systems for solid dosage forms containing acyclovir with superior biopharmaceutical properties., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study describes the treatment outcomes of preschoolers with severe mental health problems who were treated at the child psychiatric family day hospital for preschool children in Münster, Germany. The eclectic multi-modal treatment combines behavioral and psychodynamic techniques for both parents and children in various settings within an intermittent attendance structure provided by a multi-disciplinary team. This study evaluated 185 children with the Caregiver-Teacher Report Form (C-TRF/1.5-5), which was completed by therapists, and the Child Behavior Checklist (CBCL/1.5-5), which was completed by mothers, at admission and discharge. The mothers' ratings of their children were statistically adjusted for the distortion caused by their own psychopathology. After treatment, the patients showed significant improvement on the C-TRF/CBCL Total Problem score with an average Cohen's d = -0.50 based on therapists' ratings, d = -0.97 for the non-adjusted maternal ratings, and d = -0.68 for the adjusted maternal ratings. We conclude that specialized family day hospitals may successfully treat preschool psychiatric patients.

Background: Friedreich's ataxia is a rare autosomal recessive neurodegenerative disorder. Here we report cross-sectional baseline data to establish the biological and clinical characteristics for a prospective, international, European Friedreich's ataxia database registry., Methods: Within the European Friedreich's Ataxia Consortium for Translational Studies (EFACTS) framework, we assessed a cohort of patients with genetically confirmed Friedreich's ataxia. The primary outcome measure was the Scale for the Assessment and Rating of Ataxia (SARA) and secondary outcome measures were the Inventory of Non-Ataxia Signs (INAS), the performance-based coordination test Spinocerebellar Ataxia Functional Index (SCAFI), the neurocognitive phonemic verbal fluency test, and two quality-of-life measures: the activities of daily living (ADL) part of the Friedreich's Ataxia Rating Scale and EQ-5D. The Friedreich's ataxia cohort was subdivided into three groups: early disease onset (≤14 years), intermediate onset (15-24 years), and late onset (≥25 years), which were compared for clinical characteristics and outcome measures. We used linear regression analysis to estimate the annual decline of clinical outcome measures based on disease duration. This study is registered with ClinicalTrials.gov, number NCT02069509., Findings: We enrolled 592 patients with genetically confirmed Friedreich's ataxia between Sept 15, 2010, and April 30, 2013, at 11 sites in seven European countries. Age of disease onset was inversely correlated with the number of GAA repeats in the frataxin (FXN) gene: every 100 GAA repeats on the smaller repeat allele was associated with a 2·3 year (SE 0·2) earlier onset. Regression analyses showed significant estimated annual worsening of SARA (regression coefficient 0·86 points [SE 0·05], INAS (0·14 points [0·01]), SCAFI Z scores (-0·09 [0·01]), verbal fluency (-0·34 words [0·07]), and ADL (0·64 points [0·04]) during the first 25 years of disease; the regression slope for health-related quality-of-life state from EQ-5D was not significant (-0·33 points [0·18]). For SARA, the predicted annual rate of worsening was significantly higher in early-onset patients (n=354; 1·04 points [0·13]) and intermediate-onset patients (n=137; 1·17 points [0·22]) than in late-onset patients (n=100; 0·56 points [0·10])., Interpretation: The results of this cross-sectional baseline analysis of the EFACTS cohort suggest that earlier disease onset is associated with larger numbers of GAA repeats and more rapid disease progression. The differential estimated progression of ataxia symptoms related to age of onset have implications for the design of clinical trials in Friedreich's ataxia, for which SARA might be the most suitable measure to monitor disease progression., Funding: European Commission., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

From the perspective of attachment theory, insecure attachment can be seen as a key risk factor for the development of anxiety symptoms and anxiety disorders. This systematic review addresses the current state of empirical research on the relationship between attachment status and anxiety symptoms respective anxiety disorders in childhood and adolescence. 21 current international studies published between 2010 and 2014 were included in this systematic review. These studies were heterogeneous in target populations, methods and study design. The majority of studies supported the assumed correlation between insecure attachment and anxiety symptoms or anxiety disorders. These findings are more evident in studies with school-age children than with preschool children or adolescents. Furthermore, the disorganized-disoriented type of attachment seems to be a particular risk factor for the development of anxiety symptoms and anxiety disorders. Results were discussed in relation to attachment theory and with reference to the results of previous relevant reviews.

Background/aims: Chorein is a protein expressed in various cell types. Loss of function mutations of the chorein encoding gene VPS13A lead to chorea-acanthocytosis, an autosomal recessive genetic disease characterized by movement disorder and behavioral abnormalities. Recent observations revealed that chorein is a powerful regulator of actin cytoskeleton in erythrocytes, platelets, K562 and endothelial HUVEC cells., Methods: In the present study we have used Western blotting to study actin polymerization dynamics, laser scanning microscopy to evaluate in detail the role of chorein in microfilaments, microtubules and intermediate filaments cytoskeleton architecture and RT-PCR to assess gene transcription of the cytoskeletal proteins., Results: We report here powerful depolymerization of actin microfilaments both, in erythrocytes and fibroblasts isolated from chorea-acanthocytosis patients. Along those lines, morphological analysis of fibroblasts from chorea-acanthocytosis patients showed disarranged microtubular network, when compared to fibroblasts from healthy donors. Similarly, the intermediate filament networks of desmin and cytokeratins showed significantly disordered organization with clearly diminished staining in patient's fibroblasts. In line with this, RT-PCR analysis revealed significant downregulation of desmin and cytokeratin gene transcripts., Conclusion: Our results provide for the first time evidence that defective chorein is accompanied by significant structural disorganization of all cytoskeletal structures in human fibroblasts from chorea-acanthocytosis patients., (© 2015 S. Karger AG, Basel.)

With community samples, parent behavior can be assessed in standardized mother-child interaction situations with the observational instrument Lab-PAB by Wilson und Durbin (2012) with respect to five dimensions (Involvement, Positive Emotionality, Hostility, Intrusiveness and Discipline). We examined an adaptation of Lab-PAB for non-standardized free-play interaction situations (Play-PAB version) with a preschool psychiatric sample. We examined the internal consistency, interrater reliability and dimensional structure of the Play-PAB and its associations to the clinical relationship assessment scale PIR-GAS from DC:0-3R. Interaction sequences of 47 parent-child dyads who were treated in our Family Day Clinic were evaluated at admission with the Play-PAB, in addition to the PIR-GAS-rating. Each instrument was rated by two independent raters. We report means, standard deviations, internal consistencies and the interrater-reliability for each of the five Play-PAB scales. Furthermore we examine if the scale intercorrelations are reasonable in comparison to the original version. Finally, the associations to PIR-GAS are presented. The Play-PAB scales reflect sufficient variation of parenting, good internal consistencies and satisfactory interrater reliability. The adaptation shows psychometric properties that are comparable to the original version. The scale intercorrelations as well as the associations to PIR-GAS are reasonable. The Play-PAB is a promising instrument for assessing different aspects of parent behavior in a preschool psychiatric sample, with meaningful associations to parent-child-relationship quality.

Evaluation of Treatment of Mothers at the Family Day Hospital in Münster, Germany. Mothers of preschool children have limited access to mental health treatment services. The Family Day Hospital for Preschool Children at the University Hospital Münster, Germany, offers therefore a specialized treatment for mothers with their preschool children. The therapy outcome of mothers is evaluated in effectiveness study by a pre-post-design. For mothers, therapy was composed of individual session and couple sessions with the partner, video-based parent-child-interaction therapy, and parent group sessions. We evaluated the psychiatric symptom burden of N = 103 mothers at admission and discharge with the Symptom Checklist-90-Revised (SCL-90-R) above the clinical cut-off &#8805; 0.57. After treatment the mothers showed significant improvement on the global severity index (GSI) with an average Cohen's d = 1.64 (p0.001). We identified the following positively associated moderator variables of maternal improvement by a multiple regression analysis: the initial symptom burden, the educational level of the mother, not restricted housing conditions, and the age of the child. We conclude that especially distressed parents benefit from the treatment in the Family Day Hospital for Preschool Children.

The major threat in prostate cancer is the occurrence of metastases in androgen-independent tumor stage, for which no causative cure is available. Here we show that metastatic behavior of androgen-independent prostate tumor cells requires the protein-kinase-C-related kinase (PRK1/PKN1) in vitro and in vivo. PRK1 regulates cell migration and gene expression through its kinase activity, but does not affect cell proliferation. Transcriptome and interactome analyses uncover that PRK1 regulates expression of migration-relevant genes by interacting with the scaffold protein sperm-associated antigen 9 (SPAG9/JIP4). SPAG9 and PRK1 colocalize in human cancer tissue and are required for p38-phosphorylation and cell migration. Accordingly, depletion of either ETS domain-containing protein Elk-1 (ELK1), an effector of p38-signalling or p38 depletion hinders cell migration and changes expression of migration-relevant genes as observed upon PRK1-depletion. Importantly, a PRK1 inhibitor prevents metastases in mice, showing that the PRK1-pathway is a promising target to hamper prostate cancer metastases in vivo. Here we describe a novel mechanism controlling the metastatic behavior of PCa cells and identify PRK1 as a promising therapeutic target to treat androgen-independent metastatic prostate cancer.

Lysine-specific demethylase 1 (LSD1) was shown to control gene expression and cell proliferation of androgen-dependent prostate cancer (PCa) cells, whereas the role of LSD1 in androgen-independent metastatic prostate cancer remains elusive. Here, we show that depletion of LSD1 leads to increased migration and invasion of androgen-independent PCa cells. Transcriptome and cistrome analyses reveal that LSD1 regulates expression of lysophosphatidic acid receptor 6 (LPAR6) and cytoskeletal genes including the focal adhesion adaptor protein paxillin (PXN). Enhanced LPAR6 signalling upon LSD1 depletion promotes migration with concomitant phosphorylation of PXN. In mice LPAR6 overexpression enhances, whereas knockdown of LPAR6 abolishes metastasis of androgen-independent PCa cells. Taken together, we uncover a novel mechanism of how LSD1 controls metastasis and identify LPAR6 as a promising therapeutic target to treat metastatic prostate cancer.

Mutations in the gene coding for Sequestosome 1 (SQSTM1) have been genetically associated with amyotrophic lateral sclerosis (ALS) and Paget disease of bone. In the present study, we analyzed the SQSTM1 coding sequence for mutations in an extended cohort of 1,808 patients with frontotemporal lobar degeneration (FTLD), ascertained within the European Early-Onset Dementia consortium. As control dataset, we sequenced 1,625 European control individuals and analyzed whole-exome sequence data of 2,274 German individuals (total n = 3,899). Association of rare SQSTM1 mutations was calculated in a meta-analysis of 4,332 FTLD and 10,240 control alleles. We identified 25 coding variants in FTLD patients of which 10 have not been described. Fifteen mutations were absent in the control individuals (carrier frequency <0.00026) whilst the others were rare in both patients and control individuals. When pooling all variants with a minor allele frequency <0.01, an overall frequency of 3.2 % was calculated in patients. Rare variant association analysis between patients and controls showed no difference over the whole protein, but suggested that rare mutations clustering in the UBA domain of SQSTM1 may influence disease susceptibility by doubling the risk for FTLD (RR = 2.18 [95 % CI 1.24-3.85]; corrected p value = 0.042). Detailed histopathology demonstrated that mutations in SQSTM1 associate with widespread neuronal and glial phospho-TDP-43 pathology. With this study, we provide further evidence for a putative role of rare mutations in SQSTM1 in the genetic etiology of FTLD and showed that, comparable to other FTLD/ALS genes, SQSTM1 mutations are associated with TDP-43 pathology.

Purpose: A frequently used chemotherapeutic agent in hyperthermic intraperitoneal chemotherapy (HIPEC) is mitomycin C (MMC) which induces DNA damage and apoptosis in tumor cells. In addition, MMC activates DNA damage response (DDR) leading to repair mechanisms counteracting the effect of chemotherapy. COP9 signalosome (CSN) positively influences the DDR pathway by its intrinsic deneddylating and associated kinase activities. In an in vitro HIPEC model, we studied the impact of curcumin, an inhibitor of CSN-associated kinases, and of the microRNA (miRNA) let-7a-1, an inhibitor of CSN subunit expression, on the MMC-induced apoptosis in human HT29 colon cancer cells., Methods: Cells were incubated at 37 °C and indicated concentrations of MMC in a medium preheated to 42 °C as under HIPEC conditions for 1 or 4 h. HT29 cells were cotreated with 50 μM curcumin or transfected with let-7a-1 miRNA mimic. After incubation, cells were analyzed by Western blotting, densitometry, and caspase-3 ELISA., Results: An increase of CSN subunits in response to MMC treatment was detected. Apoptosis was only measured after 4 h with 50 μM MMC. MMC-induced apoptosis was elevated by cotreatment with curcumin. Transfection of HT29 cells with let-7a-1 reduced the expression of tested CSN subunits associated with the accumulation of the pro-apoptotic factors p27 and p53., Conclusions: In response to MMC treatment, the CSN is elevated as a regulator of DDR retarding apoptosis in tumor cells. The therapeutic effect of HIPEC can be increased by inhibiting CSN-associated kinases via curcumin or by blocking CSN expression with let-7a-1 miRNA.

Background: Erythropoietin (EPO) derivatives have been found to increase frataxin levels in Friedreich's ataxia (FRDA) in vitro. This multicenter, double-blind, placebo-controlled, phase II clinical trial aimed to evaluate the safety and tolerability of Lu AA24493 (carbamylated EPO; CEPO)., Methods: Thirty-six ambulatory FRDA patients harboring >400 GAA repeats were 2:1 randomly assigned to either CEPO in a fixed dose (325 µg thrice-weekly) or placebo. Safety and tolerability were assessed up to 103 days after baseline. Secondary outcome measures of efficacy (exploration of biomarkers and ataxia ratings) were performed up to 43 days after baseline., Results: All patients received six doses of study medication. Adverse events were equally distributed between CEPO and placebo. There was no evidence for immunogenicity of CEPO after multiple dosing. Biomarkers, such as frataxin, or measures for oxidative stress and ataxia ratings did not differ between CEPO and placebo., Conclusion: CEPO was safe and well tolerated in a 2-week treatment phase. Secondary outcome measures remained without apparent difference between CEPO and placebo., (© 2014 International Parkinson and Movement Disorder Society.)

Increased maternal psychopathology may bias mothers' ratings about child psychopathology. In this study we examined whether internalizing and externalizing behavior problems in young children were biased through maternal psychopathology. The clinical sample comprised 247 preschool-age patients who attended the Family Day Hospital in Münster, Germany. Internalizing and externalizing behavior problems were assessed by the CBCL/1.5-5, and maternal psychopathology was assessed by the SCL-90-R Global Severity Index (GSI). Three theoretical perspectives were tested by comparing the model fit of three structural equation models, namely the accuracy, distortion, and combinatory model. All of the models aimed to integrate multi-informant ratings from mother, therapists, and kindergarten teachers, but differed in the question which paths had to be significant. The distortion model fit the data best and supported the notion that there was a psychopathology-related bias in mothers' ratings. On the basis of this finding, we developed correction formulas comparable to Müller and Furniss (2013), in order to statistically control for this distortion. We discussed post-hoc explanations about why mothers with increased psychopathology gave higher ratings on the CBCL/1.5-5, including a better recall of internalizing symptoms, less flexible and effective parenting, and more perceived distress by child externalizing behavior., (© 2013 Published by Elsevier Ireland Ltd.)

Objective: To study the anti-herpes simplex virus (HSV) activity of a (1→6)-(1→3)-β-D-glucan isolated from Agaricus brasiliensis fruiting bodies (FR) as well as its chemically sulfated derivative (FR-S)., Methods: The antiherpetic activity and mechanism of action was studied by viral plaque assay applying different methodological strategies., Results: Although FR presented no in vitro antiherpetic action at 1 mg/ml, FR-S displayed promising anti-HSV-1 and anti-HSV-2 activities in both simultaneous and postinfection treatments, resulting in selectivity indices (CC₅₀/EC₅₀) higher than 393. FR-S had no virucidal effect, but significantly suppressed HSV-1 (EC₅₀ = 0.32 µg/ml) and HSV-2 (EC₅₀ = 0.10 µg/ml) adsorption. FR-S was less effective on adsorption inhibition of mutant virus strains devoid of gC (HSV-1 gC⁻39 and HSV-2 gCneg1), indicating a possible interaction with this glycoprotein. The reduction of viral adsorption upon cell pretreatment with FR-S also suggests its interaction with cellular components. FR-S inhibited HSV-1 (EC₅₀ = 8.39 µg/ml) and HSV-2 (EC₅₀ = 2.86 µg/ml) penetration more efficiently than heparin. FR-S reduced HSV-1 and HSV-2 cell-to-cell spread. A synergic effect between FR-S and acyclovir was also detected., Conclusions: FR-S displays an interesting mechanism of antiviral action and represents a promising candidate for the treatment and/or prevention of herpetic infections, to be used as a single therapeutic agent or in combination with acyclovir., (© 2014 S. Karger AG, Basel.)

Psychiatric treatment of children in preschool age (0-6 years) and their parents is an expanding field of research due to its high clinical significance. Specific family psychiatric treatment programs have been developed to meet the demands of this young age group, but are little known. A multiprofessional intermittent treatment approach sensitive to developmental and family context has been established in the Preschool Family Day Hospital for Infants, Toddlers and Preschoolers and their Families at Münster University Hospital, Germany. Group and individual therapeutic interventions for both children and parents, video-based parent-child-interaction therapy, psychiatric and psychotherapeutic treatments of parents and family therapeutic interventions integrating siblings are supporting and enhancing each other in an innovative and integrated family psychiatric program. First results of evaluation studies are reported that show that this treatment is effective.

The often-reported low informant agreement about child psychopathology between multiple informants has lead to various suggestions about how to address discrepant ratings. Among the factors that may lower agreement that have been discussed is informant credibility, reliability, or psychopathology, which is of interest in this paper. We tested three different models, namely, the accuracy, the distortion, and an integrated so-called combined model, that conceptualize parental ratings to assess child psychopathology. The data comprise ratings of child psychopathology from multiple informants (mother, therapist and kindergarten teacher) and ratings of maternal psychopathology. The children were patients in a preschool psychiatry unit (N=247). The results from structural equation modeling show that maternal ratings of child psychopathology were biased by maternal psychopathology (distortion model). Based on this statistical background, we suggest a method to adjust biased maternal ratings. We illustrate the maternal bias by comparing the ratings of mother to expert ratings (combined kindergarten teacher and therapist ratings) and show that the correction equation increases the agreement between maternal and expert ratings. We conclude that this approach may help to reduce misclassification of preschool children as 'clinical' on the basis of biased maternal ratings., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Chorea-acanthocytosis (ChAc), a lethal disease caused by defective chorein, is characterized by neurodegeneration and erythrocyte acanthocytosis. The functional significance of chorein in other cell types remained ill-defined. The present study revealed chorein expression in blood platelets. As compared to platelets from healthy volunteers, platelets from patients with ChAc displayed a 47% increased globular/filamentous actin ratio, indicating actin depolymerization. Moreover, phosphoinositide-3-kinase subunit p85 phosphorylation, p21 protein-activated kinase (PAK1) phosphorylation, as well as vesicle-associated membrane protein 8 (VAMP8) expression were significantly reduced in platelets from patients with ChAc (by 17, 22, and 39%, respectively) and in megakaryocytic (MEG-01) cells following chorein silencing (by 16, 54, and 11%, respectively). Activation-induced platelet secretion from dense granules (ATP release) and α granules (P-selectin exposure) were significantly less (by 55% after stimulation with 1 μg/ml CRP and by 33% after stimulation with 5 μM TRAP, respectively) in ChAc platelets than in control platelets. Furthermore, platelet aggregation following stimulation with different platelet agonists was significantly impaired. These observations reveal a completely novel function of chorein, i.e., regulation of secretion and aggregation of blood platelets.

Background: The Parent-Infant Relationship Global Assessment Scale (PIR-GAS) signifies a conceptually relevant development in the multi-axial, developmentally sensitive classification system DC:0-3R for preschool children. However, information about the reliability and validity of the PIR-GAS is rare. A review of the available empirical studies suggests that in research, PIR-GAS ratings can be based on a ten-minute videotaped interaction sequence. The qualification of raters may be very heterogeneous across studies., Methods: To test whether the use of the PIR-GAS still allows for a reliable assessment of the parent-infant relationship, our study compared a PIR-GAS ratings based on a full-information procedure across multiple settings with ratings based on a ten-minute video by two doctoral candidates of medicine. For each mother-child dyad at a family day hospital (N = 48), we obtained two video ratings and one full-information rating at admission to therapy and at discharge. This pre-post design allowed for a replication of our findings across the two measurement points. We focused on the inter-rater reliability between the video coders, as well as between the video and full-information procedure, including mean differences and correlations between the raters. Additionally, we examined aspects of the validity of video and full-information ratings based on their correlation with measures of child and maternal psychopathology., Results: Our results showed that a ten-minute video and full-information PIR-GAS ratings were not interchangeable. Most results at admission could be replicated by the data obtained at discharge. We concluded that a higher degree of standardization of the assessment procedure should increase the reliability of the PIR-GAS, and a more thorough theoretical foundation of the manual should increase its validity.

Background: An increasing number of empirical studies indicate that infants, toddlers and preschoolers may suffer from non-transient mental illnesses featuring developmental psychopathology. A few innovative child psychiatric approaches have been developed to treat infants, toddlers and preschoolers and their families, but have not yet been conceptually presented and discussed in the framework of different healthcare systems. The organizational and clinical experience gained while developing specific approaches may be important across disciplines and guide future developments in psychiatric treatment of infants, toddlers, preschoolers and their families., Results: This article introduces the Preschool Family Day Hospital for Infants, Toddlers and Preschoolers and their Families at Münster University Hospital, Germany. This hospital is unique in the German healthcare system with regard to its social-service institution division of labor. Specifically, it uses an intermittent treatment approach and an integrated interactional family psychiatric approach to treat children and their parents as separate patients. This multidisciplinary, developmentally and family-oriented approach includes components of group treatments with children and separate treatments with parents. Specific techniques include video-assisted treatments of the parent-child interaction, psychiatric and psychotherapeutic treatments for parents, and conjoint family therapies that include both parents and siblings., Conclusions: The Family Day Hospital for infants, toddlers and preschoolers and their families offers innovative family-oriented treatments for those who suffer from a wide range of severe child psychiatric disorders that cannot be sufficiently treated in outpatient settings. Treatment is based on the need for family-oriented approaches to the early psychiatric treatment of infants, toddlers and preschoolers. Family day hospitals are an innovative approach to preschool child psychiatry that requires further evaluation.

Background/aim: Interleukin-12 (IL-12) and interferon gamma (IFN-γ) are key cytokines in immunemediated equine melanoma therapy. Currently, a method for accurate simultaneous quantification of these equine cytokines is lacking. Therefore, we sought to establish an assay that allows for accurate and simultaneous quantification of equine IL-12 (eIL-12) and IFN-γ (eIFN-γ)., Materials and Methods: Several antibodies were evaluated for cross-reactivity to eIL-12 and eIFN-γ and were used to establish a bead-based Luminex assay, which was subsequently applied to quantify cytokine concentrations in biological samples., Results: Cytokine detection ranged from 31.5-5,000 pg/ml and 15-10,000 pg/ml for eIL-12 and eIFN-γ, respectively. eIL-12 was detected in supernatants of stimulated peripheral blood mononuclear cells (PBMCs) and supernatants/cell lysates of eIL-12 expression plasmid-transfected cells. Low or undetectable cytokine concentrations were measured in negative controls. In equine serum samples, the mean measured eIL-12 concentration was 1,374 ± 8 pg/ml. The bead-based assay and ELISA for eIFN-γ used to measure eIFN-γ concentrations, showed similar concentrations., Conclusion: Results demonstrate, to our knowledge for the first time, that cross-reactive antibody pairs to eIL-12 and eIFN-γ and Luminex bead-based technology allow for accurate, simultaneous and multiplexed quantification of these key cytokines in biological samples.

Streptavidin is a tetrameric protein with an extremely high affinity to biotin and different biotin-like peptide-tags. This characteristic causes its widespread use in biotechnology. Streptavidin is produced by the fermentation of wild type Streptomyces avidinii or by recombinant Streptomyces lavendulae, Escherichia coli, and Bacillus subtilis strains. However, little is known about the influence of power input and oxygen supply as well as feeding strategies on the production of streptavidin by S. avidinii. This paper provides a systematic analysis of the effect of rotary frequency of the stirrer, leading to a plateau-like streptavidin formation behaviour between 400 and 700 min(-1). This plateau was characterized by specific power inputs between 79 and 107 W L(-1) and corresponding maximal product concentrations of 6.90 μM in 6 days. Lower as well as higher rotary frequencies were not beneficial. Subsequently, a linear fed-batch procedure could be established reproducibly yielding 39.20 μM streptavidin in 14 days, characterized by a constant productivity of 114 nM h(-1). Fed-batch procedures based on dissolved oxygen were less efficient. The linear feeding strategy presented in this paper led to the highest streptavidin concentration ever reported and exceeded the maximal product level given in the literature drastically by a factor of 8.5., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We assessed the geographical distribution of C9orf72 G(4) C(2) expansions in a pan-European frontotemporal lobar degeneration (FTLD) cohort (n = 1,205), ascertained by the European Early-Onset Dementia (EOD) consortium. Next, we performed a meta-analysis of our data and that of other European studies, together 2,668 patients from 15 Western European countries. The frequency of the C9orf72 expansions in Western Europe was 9.98% in overall FTLD, with 18.52% in familial, and 6.26% in sporadic FTLD patients. Outliers were Finland and Sweden with overall frequencies of respectively 29.33% and 20.73%, but also Spain with 25.49%. In contrast, prevalence in Germany was limited to 4.82%. In addition, we studied the role of intermediate repeats (7-24 repeat units), which are strongly correlated with the risk haplotype, on disease and C9orf72 expression. In vitro reporter gene expression studies demonstrated significantly decreased transcriptional activity of C9orf72 with increasing number of normal repeat units, indicating that intermediate repeats might act as predisposing alleles and in favor of the loss-of-function disease mechanism. Further, we observed a significantly increased frequency of short indels in the GC-rich low complexity sequence adjacent to the G(4) C(2) repeat in C9orf72 expansion carriers (P < 0.001) with the most common indel creating one long contiguous imperfect G(4) C(2) repeat, which is likely more prone to replication slippage and pathological expansion., (© 2012 Wiley Periodicals, Inc.)