bluuebunny/crossref_metadata_2025 huggingface dataset:
An Optimization Approach for the Production of Fatlike Protein Isolates from Different Leguminous Seeds Using Response Surface Methodology
Authors: Sussmann D. , Halter T. , Pickardt C. , Schweiggert‐Weisz U. , Eisner P.
Published: October 2013

Plant proteins are gaining increasing importance in the food industry and are isolated by extraction followed by precipitation. However, the yields obtained are often unsatisfactory, especially when isolating proteins for specific purposes. In this study, relevant processing parameters and different raw materials have been investigated systematically to obtain high yields of protein preparations with characteristic fatlike textural properties by salt‐assisted extraction followed by dilution precipitation. A d‐optimal design was used and analyzed by response surface methodology considering individual processing parameters (independent variables) and their influence on extraction and protein yield (dependent variables). The extraction parameters (pH value [5.0–6.5], NaCl concentration [0.0–1.0 M], solid‐to‐solvent ratio [1:7–1:13]) and different raw materials (peas, lentils, white beans, faba beans, soybeans, full‐fat und deoiled lupins) were investigated. Quadratic equations for the response variables were calculated and used to identify optimum protein extraction conditions for all raw materials used. The highest protein yields (38%) were calculated for full‐fat lupins as the raw material extracted at pH 5.0, a NaCl concentration of 0.6 M and a solid‐to‐solvent ratio of 1:7. The protein yields of other grain legumes were considerably lower. Furthermore, the sensory profile of the lupin protein isolate showed unique creamy, smooth and fatlike characteristics.

Practical Applications

Plant‐based proteins, particularly protein isolates from grain legumes, are becoming increasingly important to industry as food ingredients due to their nutritional and functional properties. The major draw back of the specific protein isolation is the poor economic efficiency of the process and the low protein yields. Even though some processing parameters for protein isolation are well established, there is a lack of systematic studies on protein preparations with specific properties using different raw materials. In this study, the influence of key process parameters on the overall protein yields were investigated using pea, lentil, white bean, faba bean, soybean and lupin as raw materials. The findings will facilitate the choice of different leguminous seeds for the production of protein isolates and their potential use as fat replacers.
How to realise flood risk management plans efficiently in an urban area – the Seseke project
Authors: Johann G. , Leismann M.
Published: October 2013

This article discusses the manner in which a flood risk management plan (FRMP) can be created for an urban area that is subject to many high‐level risks. When the work processes, from the provisional risk evaluation of waters through the determination of flood plains to the creation of the FRMP, are viewed as a whole, the following questions arise: on what basis can an FRMP be created, how much time and human resources are available for this and what objectives can be achieved under these conditions? These questions, which are critical for the efficacy of an FRMP, are discussed and possible practical measures are identified.
Screening and Performance of Lactobacillus plantarum E11 with Bacteriocin‐Like Substance Secretion as Fermentation Starter of Sichuan Pickle
Authors: Rao Yu , Chang Wei , Xiang Wenliang , Li Mingyuan , Che Zhenming , Tang Jie
Published: October 2013

Bacteriocinogenic Lactobacillus plantarum E11 was selected and applied for the controlled fermentation of Sichuan pickle. By agar well diffusion assay, Lb. plantarum E11 with bacteriocin‐like substances (BLISs) production against Escherichia coli, Listeria innocua and Staphylococcus aureus was selected from eight Sichuan pickle samples. Lb. plantarum E11 could grow in Chinese leaf mustard juice media and produce BLISs with favorable antibacterial characteristics. Novel starter consisting of Lb. plantarum E11 and Leuconostoc mesenteroides CICC (China Center of Industrial Culture Collection) 21859 were applied for controlled fermentation of Chinese leaf mustard, as well as commercial starter and spontaneous fermentation. The microbial dynamics in novel starter fermentation was similar as commercial starter fermentation and Lb. plantarum E11 comprised 35.3–63.8% of the total flora. The inhibition activity of novel starter fermentation against pathogens was achieved on the fifth day, which was faster and higher than commercial and spontaneous fermentation.

Practical Applications

The natural fermentation and nonsterile process make Sichuan pickle sensitive to foodborne pathogens. The innate immunity of Sichuan pickle conferred by natural lactic acid bacteria fermentation is not strong enough to suppress the foodborne pathogens especially at the early phase of fermentation. It is necessary to develop bacteriocinogenic starter culture with wide and stable antibacterial spectrum for Sichuan pickle fermentation. This study provided a novel starter including bacteriocinogenic Lb. plantarum E11, which could dominate the Sichuan pickle fermentation as traditional commercial starter and furthermore afford the Sichuan pickle self‐protection against foodborne pathogens at the early phase of fermentation.
Value of wash‐in and wash‐out time in the diagnosis between hepatocellular carcinoma and other hepatic nodules with similar vascular pattern on contrast‐enhanced ultrasound
Authors: Kong Wen‐Tao , Wang Wen‐Ping , Huang Bei‐Jian , Ding Hong , Mao Feng
Published: October 2013

Background and Aim

The study aims to demonstrate whether the wash‐in and wash‐out time can be reliable as a criterion in the differential diagnosis between hepatocellular carcinoma (HCC) and other hepatic nodules with vascular pattern similar to HCC on contrast‐enhanced ultrasound (CEUS).

Methods

From February 2012 to February 2013, 214 patients with hepatic nodules displayed rapid hyperenhancement and quick wash‐out on CEUS were included in this study. Before performing CEUS, all nodules were examined by grayscale ultrasonography and color Doppler techniques. CEUS was performed with SonoVue and low mechanical index technique. The initial time to enhancement, time to peak, time of the nodule being hypoenhanced were comparatively studied between HCCs and other hepatic nodules.

Results

Of all the 214 nodules, 209 were malignant (164 HCCs, 31 metastases, 10 intrahepatic cholangiocarcinomas (ICCs), 3 combined hepatocellular‐cholangiocarcinomas, 1 epithelioid hemangioendothelioma), and five were benign (two inflammatory pseudotumors, one focal nodular hyperplasia nodule, one hemangioma, and one hyperplastic nodule). Metastases and ICCs showed more rapid wash‐out than HCCs (P < 0.05): 16 of 31 metastases and 7 of all ICCs showed wash‐out by 40 s after injection.

Conclusions

Some focal liver lesions can show enhancement pattern similar to HCCs on CEUS. The wash‐out time may be useful in the differential diagnosis.
Systematic review of efficacy and outcomes of salvage liver transplantation after primary hepatic resection for hepatocellular carcinoma
Authors: Chan Daniel L , Alzahrani Nayef A , Morris David L , Chua Terence C
Published: October 2013

Background and Aim

Upfront liver transplantation is the gold standard in the treatment of patients with hepatocellular carcinoma (HCC) and cirrhosis, but a shortage of donor organs negatively impacts on survival outcomes, with significant disease progression during long waiting lists. This systematic review evaluates the safety and efficacy of salvage liver transplantation (SLT) as treatment for recurrent HCC after initial hepatic resection.

Methods

Electronic searches of Pubmed, Embase, and Medline databases identified 130 abstracts, from which 16 eligible studies comprising 319 patients were selected for review. Studies adopting SLT following primary hepatic resection for recurrent HCC with more than five patients were included. Demographic details, morbidity and mortality indices, and survival outcomes were collected from each study and were tabulated.

Results

All patients included in the studies had liver cirrhosis, with the majority being Child‐Pugh A (50%) and B (33%). The etiology of liver disease was hepatitis B in the majority of patients (84%). Disease recurrence occurred in 27–80% of patients at a median of 21.4 months (range 14.5–34) following initial resection. SLTs were performed on 41% of recurrences, and were associated with biliary complications (8%), infection (11%), bleeding (8%), and vascular complications (7%). There were 18 perioperative deaths (5.6%). The median 1‐, 3‐, and 5‐year overall and disease‐free survival was 89%, 80%, and 62%, and 86%, 68%, and 67%, respectively.

Conclusion

Synthesis of available observational studies suggests that SLT following primary hepatic resection is a highly applicable strategy with long‐term survival outcomes that are comparable to upfront liver transplantation.
Model for End‐Stage Liver Disease score versus Maddrey Discriminant Function score in assessing short‐term outcome in alcoholic hepatitis
Authors: Kadian Monil , Kakkar Rajesh , Dhar Minakshi , Kaushik Rajeev Mohan
Published: October 2013

Background and Aims

The Maddrey Discriminant Function (mDF) score and the Model for End‐Stage Liver Disease (MELD) score are standard prognostic scores for predicting disease severity and mortality in alcoholic hepatitis (AH).This prospective study compared the MELD score and the mDF score as predictors of short‐term outcome in AH.

Methods

The admission MELD score and the mDF score were assessed in 47 patients with a diagnosis of AH in the Himalayan Institute Hospital, Dehradun, India and the concordance (C) statistics of the two scores for 28‐day mortality were determined and compared.

Results

Both the MELD score and the mDF score on day 1 were significantly higher in non‐survivors than in survivors (P = 0.0001 each). The C‐statistic for 28‐day mortality for the MELD score was 0.91 (P < 0.0001, 95% confidence interval [CI] 0.79–0.97) and for the mDF score 0.90 (P < 0.0001, 95% CI 0.78–0.97). There was no significant difference between the C‐statistics of the two scores (P = 0.83, 95% CI −0.07 to 0.09). For predicting 28‐day mortality, the optimal MELD score of > 19 (sensitivity 91.6% and specificity 85.7%) corresponded to the mDF score of > 52.8 (sensitivity 91.6% and specificity 82.8%).

Conclusions

Both the MELD score and the mDF score at admission were strong and equally good predictors of 28‐day mortality in patients with AH, but the optimal mDF score corresponding to optimal MELD score was higher than the conventional one. Thus, MELD score may be used as an alternative to mDF score for predicting short‐term mortality in AH with an advantage.
A clinical‐pathological analysis of hepatitis B virus recurrence after liver transplantation in Chinese patients
Authors: Gao Yin‐jie , Zhang Min , Jin Bo , Meng Fan‐ping , Ma Xue‐mei , Liu Zhen‐wen , Su Hai‐bin , Zhao Jing‐min , Li Han‐wei
Published: October 2013

Background and Aim

Liver transplantation (LT) for hepatitis B virus (HBV)‐related disease can be complicated by HBV recurrence. The aim of this study was to evaluate the risk factors, prophylaxis treatment, and histological characteristics of HBV recurrence after LT when using long‐term, low‐dose hepatitis B immunoglobulin (HBIG) plus nucleoside analog (lamivudine [LAM] or entecavir [ETV]).

Methods

Retrospective data from 253 adult LT patients using long‐term, low‐dose HBIG plus nucleoside analog after LT, for a mean treatment duration of 1–72 months, were collected from a single center in Beijing, China. Univariate analyses were conducted to determine the association among gender, age, hepatocellular carcinoma, hepatitis B e antigen‐positive status, HBV‐DNA level and tyrosine‐methionine‐aspartate‐aspartate (YMDD) mutations on HBV recurrence in these patients.

Results

Overall, the HBV recurrence rate was 6.32% (16/253). There was no significant difference in the survival rate between the HBV recurrence and non‐recurrence groups. Risk factors for HBV recurrence were: hepatitis B e antigen positivity, HBV‐DNA > 105 copies/mL, hepatocellular carcinoma, and YMDD mutation. Sixteen patients receiving LAM had HBV recurrence (16/169; mean treatment duration: 61.8 ± 18.3 months). No HBV recurrence occurred in patients receiving ETV after LT (0/84; mean treatment duration: 57.1 ± 15.9 months). Differences in rate of mortality and HBV recurrence were not significant between the two groups.

Conclusions

LT is an effective treatment for HBV‐related end‐stage liver disease. The combination of ETV and intramuscular HBIG for HBV recurrence prophylaxis after LT was more effective than LAM, especially in Chinese patients with HBV recurrence risk factors.
Prevalence of Vitamin D Deficiency and Association with Functional Status in Newly Admitted Male Veteran Nursing Home Residents
Authors: Kojima Gotaro , Tamai Anna , Masaki Kamal , Gatchell Gregory , Epure James , China Craig , Ross G. Webster , Petrovitch Helen , Tanabe Marianne
Published: October 2013

Objectives

To provide the first report on prevalence of vitamin D deficiency in newly admitted nursing home (NH) residents and associations with functional disabilities and chronic diseases.

Design

Retrospective chart review.

Setting

Nursing home (NH).

Participants

Male veterans newly admitted to a NH for rehabilitation, skilled‐nursing care, intermediate care, or respite care between January 2011 and June 2012.

Measurements

Total serum 25‐hydroxyvitamin D (25(OH)D) levels were measured on admission. Vitamin D supplement users and those without 25(OH)D measurement within 7 days of admission were excluded, leaving an analytical sample of 104 residents. Vitamin D deficiency was defined as 25(OH)D less than 20 ng/mL. Data were collected on age, ethnicity, season, body mass index (BMI), functional disability in activities of daily living (ADLs) (mobility, bathing, dressing, toileting, continence, and feeding), and prevalent chronic diseases.

Results

Prevalence of vitamin D deficiency was 49.0%. In multivariate logistic regression models adjusted for age, ethnicity, and BMI, vitamin D deficiency was significantly associated with number of ADL disabilities (odds ratio (OR) = 1.4 for each 1‐point increase in ADL disability score, P = .03) and prevalent diabetes mellitus (OR = 3.0, P = .03). In regression models using each ADL disability as a separate variable, only disability in feeding (OR = 4.7, P = .05) and diabetes mellitus (OR = 2.9, P = .04) remained significant.

Conclusion

Almost half the individuals entering the NH and not taking vitamin D supplements had vitamin D deficiency. Greater number of ADL disabilities, disability in feeding, and prevalent diabetes mellitus were independently associated with vitamin D deficiency.
What do the terms wellness and wellbeing mean in dietary practice: an exploratory qualitative study examining women's perceptions
Authors: McMahon A. T. , O'Shea J. , Tapsell L. , Williams P.
Published: October 2013

Background

Wellness and wellbeing are terms associated with health within dietetic discourse. More broadly, these terms are found in social discourse as represented in food and nutrition consumer communications. With the increasing requirement for evidence‐based healthcare, there is an imperative to understand whether these terms are meaningful to individuals typically targeted for nutrition interventions and whether there are any implications for dietetic education.

Methods

To explore the understanding of these terms, eight semi‐structured focus groups were conducted with 32 female participants (age range 23–79 years) who were actively engaged in managing their health. Overall understanding of the terms, factors that impacted perceptions and any relationships with food behaviour were investigated with the groups. Group discussions were transcribed verbatim and each transcript was examined by two researchers. Inductive analysis linking codes into main thematic categories was conducted using the constant comparison approach across the full data set.

Results

Wellness and wellbeing were identified as meaningful terms associated with health. A theoretical framework of wellness and wellbeing reflecting these meanings was developed linking four dominant thematic areas. These were Desired outcomes (most sought after result); Taking control (self management strategies); Internal influences (various personal inner factors influencing behaviours); and External influences (plethora of peripheral factors influencing behaviours).

Conclusions

Wellness and wellbeing are terms that are relevant and aspirational for individuals typically targeted for nutrition intervention. A theoretical framework of dominant areas of influence on notions of wellness and wellbeing was identified. This theoretical framework is worthy of further research to determine usefulness and effectiveness in dietetic practice settings.
Development of a quantitative food frequency questionnaire for use among rural South Africans in KwaZulu‐Natal
Authors: Sheehy T. , Kolahdooz F. , Mtshali T. L. , Khamis T. , Sharma S.
Published: October 2013

Background

South Africa is experiencing a dietary and lifestyle transition as well as increased rates of noncommunicable chronic diseases. Limited information is available on the diets of rural populations. The present study aimed to characterise the diets of men and women from rural KwaZulu‐Natal (KZN) and develop a quantitative food‐frequency questionnaire (QFFQ) specific for this population.

Methods

A cross‐sectional study was carried out by collecting single 24‐h dietary recalls from 81 adults and developing a QFFQ in Empangeni, KZN, South Africa.

Results

The diet of this population was limited in variety, high in plant‐based foods (especially cereals and beans), and low in animal products, vegetables and fruits. Amaize meal staple (Phutu) was consumed by over 80% of subjects and accounted for almost 45% of energy intake, as well as making an important contribution to fat and protein intake. Most of the protein consumed by the study population was plant‐based protein, with almost 40% being obtained from the consumption of phutu and beans. A culturally appropriate QFFQ was developed that includes 71 food and drink items, of which 16 are composite dishes unique to this population.

Conclusions

Once validated, this QFFQ can be used to monitor diet‐disease associations, evaluate nutritional interventions and investigate dietary changes in this population.
Substance Flow Analysis of Wastes Containing Polybrominated Diphenyl Ethers
Authors: Vyzinkarova Dana , Brunner Paul H.
Published: October 2013

The present article examines flows and stocks of Stockholm Convention regulated pollutants, commercial penta‐ and octabrominated diphenyl ether (cPentaBDE, cOctaBDE), on a city level. The goals are to (1) identify sources, pathways, and sinks of these compounds in the city of Vienna, (2) determine the fractions that reach final sinks, and (3) develop recommendations for waste management to ensure their minimum recycling and maximum transfer to appropriate final sinks. By means of substance flow analysis (SFA) and scenario analysis, it was found that the key flows of cPentaBDE stem from construction materials. Therefore, end‐of‐life (EOL) plastic materials used for construction must be separated and properly treated, for example, in a state‐of‐the‐art municipal solid waste (MSW) incinerator. In the case of cOctaBDE, the main flows are waste electrical and electronic equipment (WEEE) and, possibly, vehicles. Most EOL vehicles are exported from Vienna and pose a continental, rather than a local, problem. According to the modeling, approximately 73% of cOctaBDE reached the final sink MSW incinerator, and 17% returned back to consumption by recycling. Secondary plastics, made from WEEE, may thus contain significant amounts of cOctaBDE; however, uncertainties are high. According to uncertainty analysis, the major cause is the lack of reliable values regarding cOctaBDE concentrations in European WEEE categories 3 and 4, including cathode ray tube monitors for computers and televisions. We recommend establishing a new, goal‐oriented data set by additional analyses of waste constituents and plastic recycling samples, as well as establishing reliable mass balances of polybrominated diphenyl ethers’ flows and stocks by means of SFA.
An Exploration of the Relationship between Socioeconomic and Well‐Being Variables and Household Greenhouse Gas Emissions
Authors: Wilson Jeffrey , Tyedmers Peter , Spinney Jamie E. L.
Published: October 2013

This research reports on a multivariate analysis that examined the relationship between direct greenhouse gas (GHG) emissions and socioeconomic and well‐being variables for 1,920 respondents living in Halifax Regional Municipality, Nova Scotia, Canada, using results from the Halifax Space‐Time Activity Research Project. The unique data set allows us to estimate direct GHG emissions with an unprecedented level of specificity based on household energy use survey data and geographic positioning system–verified personal travel data. Of the variables analyzed, household size, income, community zone, age, and marital status are all statistically significant predictors of direct GHG emissions. Birthplace, ethnicity, educational attainment, perceptions of health, life satisfaction, job satisfaction, happiness, volunteering, or community belonging did not seem to matter. In addition, we examined whether those reporting energy‐efficient behaviors had lower GHG emissions. No significant differences were discovered among the groups analyzed, supporting a growing body of research indicating a disconnect between environmental attitudes and behaviors and environmental impact. Among the predictor variables, those reporting to be married, young, low income, and living in households with more people have correspondingly lower direct GHG emissions than other categories in respective groupings. Our finding that respondents with lifestyles that generate higher GHG emissions did not report to be healthier, happier, or more connected to their communities suggest that individuals can experience similar degrees of well‐being regardless of the amount of GHG emissions associated with his or her respective lifestyle.
Quantifying relationships between rooting traits and water uptake under drought in Mediterranean barley and durum wheat
Authors: Carvalho Pedro , Azam‐Ali Sayed , Foulkes M. John
Published: October 2013

In Mediterranean regions drought is the major factor limiting spring barley and durum wheat grain yields. This study aimed to compare spring barley and durum wheat root and shoot responses to drought and quantify relationships between root traits and water uptake under terminal drought. One spring barley (Hordeum vulgare L. cv. Rum) and two durum wheat Mediterranean cultivars (Triticum turgidum L. var durum cvs Hourani and Karim) were examined in soil‐column experiments under well watered and drought conditions. Root system architecture traits, water uptake, and plant growth were measured. Barley aerial biomass and grain yields were higher than for durum wheat cultivars in well watered conditions. Drought decreased grain yield more for barley (47%) than durum wheat (30%, Hourani). Root‐to‐shoot dry matter ratio increased for durum wheat under drought but not for barley, and root weight increased for wheat in response to drought but decreased for barley. The critical root length density (RLD) and root volume density (RVD) for 90% available water capture for wheat were similar to (cv. Hourani) or lower than (cv. Karim) for barley depending on wheat cultivar. For both species, RVD accounted for a slightly higher proportion of phenotypic variation in water uptake under drought than RLD.
Genome‐wide analysis and molecular dissection of the SPL gene family in Salvia miltiorrhiza
Authors: Zhang Linsu , Wu Bin , Zhao Degang , Li Caili , Shao Fenjuan , Lu Shanfa
Published: October 2013

SQUAMOSA promoter binding protein‐likes (SPLs) are plant‐specific transcription factors playing vital regulatory roles in plant growth and development. There is no information about SPLs in Salvia miltiorrhiza (Danshen), a significant medicinal plant widely used in Traditional Chinese medicine (TCM) for >1,700 years and an emerging model plant for TCM studies. Through genome‐wide identification and subsequent molecular cloning, we identified a total 15 SmSPLs with divergent sequence features, gene structures, and motifs. Comparative analysis showed sequence conservation between SmSPLs and their Arabidopsis counterparts. A phylogenetic tree clusters SmSPLs into six groups. Many of the motifs identified commonly exist in a group/subgroup, implying their functional redundancy. Eight SmSPLs were predicted and experimentally validated to be targets of miR156/157. SmSPLs were differentially expressed in various tissues of S. milltiorrhiza. The expression of miR156/157‐targeted SmSPLs was increased with the maturation of S. miltiorrhiza, whereas the expression of miR156/157 was decreased, confirming the regulatory roles of miR156/157 in SmSPLs and suggesting the functions of SmSPLs in S. miltiorrhiza development. The expression of miR156/157 was negatively correlated with miR172 during the maturation of S. miltiorrhiza. The results indicate the significance and complexity of SmSPL‐, miR156‐, and miR172‐mediated regulation of developmental timing in S. miltiorrhiza.
Prevalence of aggressive challenging behaviours in intellectual disability and its relationship to personality status: Jamaican study
Authors: Tyrer P. , Oliver P. , Tarabi S. A.
Published: October 2013

Background

Both the classification of personality disorder in intellectual disability (ID) and its identification in practice are deemed to be difficult. A simpler approach to classification and its relationship to challenging behaviours were tested in an adult Jamaican population with ID.

Method

The study was carried out in Kingston, Jamaica, as part of a programme of field trials to determine the utility of the proposed revision of personality disorders in the 11th Revision of the International Classification of Diseases (ICD‐11), in a population of adults with ID living with their families or in supported care homes. Thirty‐eight people with borderline (n = 5), mild (n = 16), moderate (n = 14) and severe (n = 3) ID were assessed at face‐to‐face interview and with relatives or staff using the provisional criteria for severity of personality disorder and its associated domain traits, and challenging behaviour was assessed using the Problem Behaviour Check List (PBCL) (a 5‐point, 7‐item scale).

Results

Using the severity scale 18 patients (47%) had no personality disorder, 7 (18%) had personality difficulty, 9 (24%) had mild personality disorder, and 4 (11%) had moderate personality disorder. None of the sample had severe personality disorder in which there is high risk of harm to self or others. Of the four major trait domains, provisionally named anankastic, detached, emotional and dissocial, three were evenly distributed in those with personality disturbance with the antagonistic (antisocial) trait less commonly shown (6 only). Scores on the PBCL were higher in those with increasing severity of personality disorder (P = 0.03) and those in the antagonistic personality trait domain had the highest PCL scores.

Conclusions

Despite previous difficulties in assessing personality disorder in intellectual difficulties the ICD‐11 classification was easy to administer in practice in this population, and the higher problem behaviour scores in those with greater severity of personality disturbance support its construct validity.
Assessing Play‐Based Activities, Child Talk, and Single Session Outcome in Family Therapy with Young Children
Authors: Willis Amber B. , Walters Lynda H. , Crane D. Russell
Published: October 2013

This exploratory, observational study was designed to reveal descriptive information regarding therapists' actual practices with preschool‐ and school‐aged children in a single session of family therapy and to investigate change mechanisms in family play therapy that have been proposed to make this approach effective. A purposive sample of 30 families receiving family therapy was recruited and video‐taped during a family session where at least one child between the ages of 4 and 12 was present. Following the session, the therapist and parent(s) completed questionnaires while one of the children (aged 4–12) was interviewed. Session recordings were coded, minute‐by‐minute, for participant talk time, visual aids or props used, and therapy technique type (e.g., play‐based/activity vs. talk‐only techniques). Hierarchical regression and canonical correlational analyses revealed evidence supporting the theory that play‐based techniques promote young children's participation, enhance the quality of the child–therapist relationship, and build positive emotional experiences in family therapy.
Time‐dependent reduction of structural complexity of the buccal epithelial cell nuclei after treatment with silver nanoparticles
Authors: PANTIC I. , PAUNOVIC J. , PEROVIC M. , CATTANI C. , PANTIC S. , SUZIC S. , NESIC D. , BASTA‐JOVANOVIC G.
Published: October 2013

Recent studies have suggested that silver nanoparticles (AgNPs) may affect cell DNA structure in in vitro conditions. In this paper, we present the results indicating that AgNPs change nuclear complexity properties in isolated human epithelial buccal cells in a time‐dependent manner. Epithelial buccal cells were plated in special tissue culture chamber / slides and were kept at 37°C in an RPMI 1640 cell culture medium supplemented with L‐glutamine. The cells were treated with colloidal silver nanoparticles suspended in RPMI 1640 medium at the concentration 15 mg L−1. Digital micrographs of the cell nuclei in a sample of 30 cells were created at five different time steps: before the treatment (controls), immediately after the treatment, as well as 15 , 30 and 60 min after the treatment with AgNPs. For each nuclear structure, values of fractal dimension, lacunarity, circularity, as well as parameters of grey level co‐occurrence matrix (GLCM) texture, were determined. The results indicate time‐dependent reduction of structural complexity in the cell nuclei after the contact with AgNPs. These findings further suggest that AgNPs, at concentrations present in today's over‐the‐counter drug products, might have significant effects on the cell genetic material.
A conserved sequence in calmodulin regulated spectrin‐associated protein 1 links its interaction with spectrin and calmodulin to neurite outgrowth
Authors: King Mikayala D. A. , Phillips Gareth W. , Bignone Paola A. , Hayes Nandini V. L. , Pinder Jennifer C. , Baines Anthony J.
Published: October 2013

Calmodulin regulated spectrin‐associated protein 1 (CAMSAP1) is a vertebrate microtubule‐binding protein, and a representative of a family of cytoskeletal proteins that arose with animals. We reported previously that the central region of the protein, which contains no recognized functional domain, inhibited neurite outgrowth when over‐expressed in PC12 cells [Baines et al., Mol. Biol. Evol. 26 (2009), p. 2005]. The CKK domain (DUF1781) binds microtubules and defines the CAMSAP/ssp4 family of animal proteins (Baines et al. 2009). In the central region, three short well‐conserved regions are characteristic of CAMSAP‐family members. One of these, CAMSAP‐conserved region 1 (CC1), bound to both βIIΣ1‐spectrin and Ca2+/calmodulin in vitro. The binding of Ca2+/calmodulin inhibited spectrin binding. Transient expression of CC1 in PC12 cells inhibited neurite outgrowth. siRNA knockdown of CAMSAP1 inhibited neurite outgrowth in PC12 cells or primary cerebellar granule cells: this could be rescued in PC12 cells by wild‐type CAMSAP1‐enhanced green fluorescent protein, but not by a CC1 mutant. We conclude that CC1 represents a functional region of CAMSAP1, which links spectrin‐binding to neurite outgrowth.

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Knockdown of the cytoskeletal protein CAMSAP1 using siRNA inhibited NGF‐induced (nerve growth factor) neurite outgrowth from PC12 cells, and axon production by cerebellar granule cells in culture. This activity is linked to a spectrin‐ and Ca2+/calmodulin‐binding region (CC1), since over‐expression of isolated CC1 inhibited neurite production from PC12 cells. We previously showed that CAMSAP1 binds microtubules at the C‐terminal CKK domain. Our data indicates CAMSAP1 is a cytoskeletal interconnector required for neurite and axon production.
CaMKII represses transcriptionally active β‐catenin to mediate acute ethanol neurodegeneration and can phosphorylate β‐catenin
Authors: Flentke George R. , Garic Ana , Hernandez Marcos , Smith Susan M.
Published: October 2013

Prenatal ethanol exposure causes persistent neurodevelopmental deficits by inducing apoptosis within neuronal progenitors including the neural crest. The cellular signaling events underlying this apoptosis are unclear. Using an established chick embryo model, we previously identified ethanol's activation of calmodulin‐dependent protein kinase II (CaMKII) as a crucial early step in this pathway. Here, we report that CaMKII is pro‐apoptotic because it mediates the loss of transcriptionally active β‐catenin, which normally provides trophic support to these cells. β‐catenin over‐expression normalized cell survival in ethanol's presence. CaMKII inhibition similarly restored β‐catenin content and transcriptional activity within ethanol‐treated cells and prevented their cell death. In contrast, inhibition of alternative effectors known to destabilize β‐catenin, including glycogen synthase kinase‐3β, Protein Kinase C, JNK, and calpain, failed to normalize cell survival and β‐catenin activity in ethanol's presence. Importantly, we found that purified CaMKII can directly phosphorylate β‐catenin. Using targeted mutagenesis we identified CaMKII phosphorylation sites within human β‐catenin at T332, T472, and S552. This is the first demonstration that β‐catenin is a phosphorylation target of CaMKII and represents a novel mechanism by which calcium signals could regulate β‐catenin‐dependent transcription. These results inform ethanol's neurotoxicity and offer unexpected insights into other neurodevelopmental and neurodegenerative disorders having dysregulated calcium or β‐catenin signaling.

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We proposed that ethanol initiates apoptosis in early neuroprogenitors through an intracellular calcium transient that activates CaMKII, which in turn destabilizes transcriptionally‐active beta‐catenin, perhaps through direct phosphorylation at Thr332, Thr472 and Ser552. This represents a novel mechanism of calcium‐modulation of Wnt signaling and of ethanol's neurotoxicity.
The secretogranin II gene is a signal integrator of glutamate and dopamine inputs
Authors: Iwase Katsuro , Ishihara Akinori , Yoshimura Shuntaro , Andoh Yoshio , Kato Masaki , Seki Naohiko , Matsumoto Eriko , Hiwasa Takaki , Muller Dominique , Fukunaga Kohji , Takiguchi Masaki
Published: October 2013

Cooperative gene regulation by different neurotransmitters likely underlies the long‐term forms of associative learning and memory, but this mechanism largely remains to be elucidated. Following cDNA microarray analysis for genes regulated by Ca2+ or cAMP, we found that the secretogranin II gene (Scg2) was cooperatively activated by glutamate and dopamine in primary cultured mouse hippocampal neurons. The Ca2+ chelator 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N’,N’‐tetraacetic acid acetoxymethyl ester (BAPTA‐AM) and the mitogen‐activated protein kinase (MAPK) kinase (MEK) inhibitor PD98059 prevented Scg2 activation by glutamate or dopamine; thus, the Ca2+/MEK pathway is predicted to include a convergence point(s) of glutamatergic and dopaminergic signaling. Unexpectedly, the protein kinase A inhibitor KT5720 enhanced Scg2 activation by dopamine. The protein‐synthesis inhibitor cycloheximide also enhanced Scg2 activation, and the proteasome inhibitor ZLLLH diminished the KT5720‐mediated augmentation of Scg2 activation. These results are concordant with the notion that dopaminergic input leads to accumulation of a KT5720‐sensitive transcriptional repressor, which is short‐lived because of rapid degradation by proteasomes. This repression pathway may effectively limit the time window permissive to Scg2 activation by in‐phase glutamate and dopamine inputs via the Ca2+/MEK pathway. We propose that the regulatory system of Scg2 expression is equipped with machinery that is refined for the signal integration of in‐phase synaptic inputs.

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We proposed hypothetical mechanism for the regulation of the secretogranin II gene as a signal integrator of glutamate and dopamine inputs. Glutamate or dopamine activates the Ca2+/MEK/ERK pathway, which thus contributes to the signal integration. Concurrently, activation of the PKA inhibitor KT5720‐sensitive pathway by dopamine leads to accumulation of the repressor protein X that is otherwise susceptible to proteasome degradation. This repression system may determine the time window permissive to the cooperative activation by in‐phase glutamate and dopamine inputs.
EphrinA/EphA‐induced ectodomain shedding of neural cell adhesion molecule regulates growth cone repulsion through ADAM10 metalloprotease
Authors: Brennaman Leann H. , Moss Marcia L. , Maness Patricia F.
Published: October 2013

EphrinA/EphA‐dependent axon repulsion is crucial for synaptic targeting in developing neurons but downstream molecular mechanisms remain obscure. Here, it is shown that ephrinA5/EphA3 triggers proteolysis of the neural cell adhesion molecule (NCAM) by the metalloprotease a disintegrin and metalloprotease (ADAM)10 to promote growth cone collapse in neurons from mouse neocortex. EphrinA5 induced ADAM10 activity to promote ectodomain shedding of polysialic acid‐NCAM in cortical neuron cultures, releasing a ~ 250 kDa soluble fragment consisting of most of its extracellular region. NCAM shedding was dependent on ADAM10 and EphA3 kinase activity as shown in HEK293T cells transfected with dominant negative ADAM10 and kinase‐inactive EphA3 (K653R) mutants. Purified ADAM10 cleaved NCAM at a sequence within the E‐F loop of the second fibronectin type III domain (Leu671‐Lys672/Ser673‐Leu674) identified by mass spectrometry. Mutations of NCAM within the ADAM10 cleavage sequence prevented EphA3‐induced shedding of NCAM in HEK293T cells. EphrinA5‐induced growth cone collapse was dependent on ADAM10 activity, was inhibited in cortical cultures from NCAM null mice, and was rescued by WT but not ADAM10 cleavage site mutants of NCAM. Regulated proteolysis of NCAM through the ephrin5/EphA3/ADAM10 mechanism likely impacts synapse development, and may lead to excess NCAM shedding when disrupted, as implicated in neurodevelopmental disorders such as schizophrenia.

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PSA‐NCAM and ephrinA/EphA3 coordinately regulate inhibitory synapse development. Here, we have found that ephrinA5 stimulates EphA3 kinase and ADAM10 activity to promote PSA‐NCAM cleavage at a site in its second FNIII repeat, which regulates ephrinA5‐induced growth cone collapse in GABAergic and non‐GABAergic neurons. These findings identify a new regulatory mechanism which may contribute to inhibitory connectivity.
Cathepsin E generates a sumoylated intracellular fragment of the cell adhesion molecule L1 to promote neuronal and Schwann cell migration as well as myelination
Authors: Lutz David , Wolters‐Eisfeld Gerrit , Schachner Melitta , Kleene Ralf
Published: October 2013

The cell adhesion molecule L1 regulates cellular responses in the developing and adult nervous system. Here, we show that stimulation of cultured mouse cerebellar neurons by a function‐triggering L1 antibody leads to cathepsin E‐mediated generation of a sumoylated 30 kDa L1 fragment (L1‐30) and to import of L1‐30 into the nucleus. Mutation of the sumoylation site at K1172 or the cathepsin E cleavage site at E1167 abolishes generation of L1‐30, while mutation of the nuclear localization signal at K1147 prevents nuclear import of L1‐30. Moreover, the aspartyl protease inhibitor pepstatin impairs the generation of L1‐30 and inhibits L1‐induced migration of cerebellar neurons and Schwann cells as well as L1‐dependent in vitro myelination on axons of dorsal root ganglion neurons by Schwann cells. L1‐stimulated migration of HEK293 cells expressing L1 with mutated cathepsin E cleavage site is diminished in comparison to migration of cells expressing non‐mutated L1. In addition, L1‐stimulated migration of HEK293 cells expressing non‐mutated L1 is also abolished upon knock‐down of cathepsin E expression and enhanced by over‐expression of cathepsin E. The findings of the present study indicate that generation and nuclear import of L1‐30 regulate neuronal and Schwann cell migration as well as myelination.

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Cell adhesion molecule L1 regulates cellular responses in the developing and adult nervous system. L1 stimulation triggers sumoylation and cleavage of L1, thus generating the L1‐70 fragment (1) which is cleaved by cathepsin E (2) yielding the L1‐30 fragment that is imported to the nucleus (3), may bind to DNA and/or nuclear proteins (4), to regulate diverse cellular functions.
GABAA receptor transmembrane amino acids are critical for alcohol action: disulfide cross‐linking and alkyl methanethiosulfonate labeling reveal relative location of binding sites
Authors: Borghese Cecilia M. , Hicks Jessica A. , Lapid Daniel J. , Trudell James R. , Harris R. Adron
Published: October 2013

Alcohols and inhaled anesthetics modulate GABAA receptor (GABAAR) function via putative binding sites within the transmembrane regions. The relative position of the amino acids lining these sites could be either inter‐ or intra‐subunit. We introduced cysteines in relevant TM locations and tested the proximity of cysteine pairs using oxidizing and reducing agents to induce or break disulfide bridges between cysteines, and thus change GABA‐mediated currents in wild‐type and mutant α1β2γ2 GABAARs expressed in Xenopus laevis oocytes. We tested for: (i) inter‐subunit cross‐linking: a cysteine located in α1TM1 [either α1(Q229C) or α1(L232C)] was paired with a cysteine in different positions of β2TM2 and TM3; (ii) intra‐subunit cross‐linking: a cysteine located either in β2TM1 [β2(T225C)] or in TM2 [β2(N265C)] was paired with a cysteine in different locations along β2TM3. Three inter‐subunit cysteine pairs and four intra‐subunits cross‐linked. In three intra‐subunit cysteine combinations, the alcohol effect was reduced by oxidizing agents, suggesting intra‐subunit alcohol binding. We conclude that the structure of the alcohol binding site changes during activation and that potentiation or inhibition by binding at inter‐ or intra‐subunit sites is determined by the specific receptor and ligand.

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Alcohols modulate GABAA receptor function. The relative location of alcohol binding pockets in transmembrane domains was assessed by substituting critical amino acids with cysteines, and testing GABA responses and alcohol modulation before and after crosslinking double cysteine mutants and labeling single cysteine mutants. Lines show absence of crosslinking (red), and crosslinking that modified GABA responses (black) and alcohol modulation (grey).
Blocking the apolipoprotein E/Amyloid β interaction in triple transgenic mice ameliorates Alzheimer's disease related amyloid β and tau pathology
Authors: Liu Shan , Breitbart Ariel , Sun Yanjie , Mehta Pankaj D. , Boutajangout Allal , Scholtzova Henrieta , Wisniewski Thomas
Published: October 2013

Inheritance of the apolipoprotein E4 (apoE4) genotype has been identified as the major genetic risk factor for late‐onset Alzheimer's disease (AD). Studies have shown that the binding between apoE and amyloid‐β (Aβ) peptides occurs at residues 244–272 of apoE and residues 12–28 of Aβ. ApoE4 has been implicated in promoting Aβ deposition and impairing clearance of Aβ. We hypothesized that blocking the apoE/Aβ interaction would serve as an effective new approach to AD therapy. We have previously shown that treatment with Aβ12‐28P can reduce amyloid plaques in APP/PS1 transgenic (Tg) mice and vascular amyloid in TgSwDI mice with congophilic amyloid angiopathy. In the present study, we investigated whether the Aβ12‐28P elicits a therapeutic effect on tau‐related pathology in addition to amyloid pathology using old triple transgenic AD mice (3xTg, with PS1M146V, APPSwe and tauP30IL transgenes) with established pathology from the ages of 21 to 26 months. We show that treatment with Aβ12‐28P substantially reduces tau pathology both immunohistochemically and biochemically, as well as reducing the amyloid burden and suppressing the activation of astrocytes and microglia. These affects correlate with a behavioral amelioration in the treated Tg mice.

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The apolipoprotein E (apoE)/amyloid beta (Aβ) interaction plays an important role in the aggregation and clearance of Aβ. We show that the apoE/Aβ binding blocker, Aβ12‐28P, elicits a therapeutic effect on tau related pathology, in addition to amyloid deposition, in 3xTg Alzheimer's disease model mice. These affects correlate with a behavioral amelioration in the treated Tg mice. We believe this presents a novel therapeutic approach for AD.
Annexin A1 Complex Mediates Oxytocin Vesicle Transport
Authors: Makani V. , Sultana R. , Sie K. S. , Orjiako D. , Tatangelo M. , Dowling A. , Cai J. , Pierce W. , Butterfield D. A. , Hill J. , Park J.
Published: October 2013

Oxytocin is a major neuropeptide that modulates the brain functions involved in social behaviour and interaction. Despite of the importance of oxytocin for the neural control of social behaviour, little is known about the molecular mechanism(s) by which oxytocin secretion in the brain is regulated. Pro‐oxytocin is synthesised in the cell bodies of hypothalamic neurones in the supraoptic and paraventricular nuclei and processed to a 9‐amino‐acid mature form during post‐Golgi transport to the secretion sites at the axon terminals and somatodendritic regions. Oxytocin secreted from the somatodendritic regions diffuses throughout the hypothalamus and its neighbouring brain regions. Some oxytocin‐positive axons innervate and secrete oxytocin to the brain regions distal to the hypothalamus. Brain oxytocin binds to its receptors in the brain regions involved in social behaviour. Oxytocin is also secreted from the axon terminal at the posterior pituitary gland into the blood circulation. We have discovered a new molecular complex consisting of annexin A1 (ANXA1), A‐kinase anchor protein 150 (AKAP150) and microtubule motor that controls the distribution of oxytocin vesicles between the axon and the cell body in a protein kinase A (PKA)‐ and protein kinase C (PKC)‐sensitive manner. ANXA1 showed significant co‐localisation with oxytocin vesicles. Activation of PKA enhanced the association of kinesin‐2 with ANXA1, thus increasing the axon‐localisation of oxytocin vesicles. Conversely, activation of PKC decreased the binding of kinesin‐2 to ANXA1, thus attenuating the axon‐localisation of oxytocin vesicles. The result of the present study suggest that ANXA1 complex coordinates the actions of PKA and PKC to control the distribution of oxytocin vesicles between the axon and the cell body.