bluuebunny/crossref_metadata_2025 huggingface dataset:
Chinese children with autism: A multiple chemical elements profile in erythrocytes
Authors: Wu Jing , Liu Duo‐Jian , Shou Xiao‐Jing , Zhang Ji‐Shui , Meng Fan‐Chao , Liu Ya‐Qiong , Han Song‐Ping , Zhang Rong , Jia Jin‐Zhu , Wang Jing‐Yu , Han Ji‐Sheng
Published: March 2018

Several lines of evidence suggested that abnormal levels of certain chemical elements may contribute to the development of autism spectrum disorders (ASD). The present work aimed to investigate the multiple chemical elements profile in the erythrocytes of autistic versus typically developing children (TDC) of China. Analyses were carried out to explore the possible association between levels of elements and the risk as well as the severity of ASD. Erythrocyte levels of 11 elements (32%) among 34 detected elements in autistic group were significantly different from those in the TDC group. To our knowledge, this is the first study which compared the levels of rare earth elements in erythrocytes between children with or without ASD. Five elements including Pb, Na, Ca, Sb, and La are associated with the Childhood Autism Rating Scale (CARS) total score. Also, a series of tendencies were found in this research which was believed to affect auditory response, taste, smell, and touch, as well as fear or nervousness. It can be concluded that Chinese autistic children suffer from multi‐chemical element imbalances which involves a complex combination of genetic and environmental factors. The results showed a significant correlation between abnormal levels of several chemical elements and the severity of the autistic syndrome.

Lay Summary

It is suggested that abnormal levels of some chemical elements may contribute to the development of autism spectrum disorders (ASD). In this work, the impact of element imbalances on the risk and severity of ASD was investigated, focusing on the analysis of abnormal levels of the multi‐chemical elements profile in erythrocytes compared with typically developing children. Furthermore, the results showed a significant correlation between abnormal levels of several chemical elements and the severity of the autistic syndrome.Autism Res2018, 11: 834–845. © 2018 International Society for Autism Research, Wiley Periodicals, Inc.
DNA Repair: The Search for Homology
Authors: Haber James E.
Published: March 2018

The repair of chromosomal double‐strand breaks (DSBs) by homologous recombination is essential to maintain genome integrity. The key step in DSB repair is the RecA/Rad51‐mediated process to match sequences at the broken end to homologous donor sequences that can be used as a template to repair the lesion. Here, in reviewing research about DSB repair, I consider the many factors that appear to play important roles in the successful search for homology by several homologous recombination mechanisms. See also the video abstract here: https://youtu.be/vm7-X5uIzS8.
Demarcation of Viral Shelters Results in Destruction by Membranolytic GTPases: Antiviral Function of Autophagy Proteins and Interferon‐Inducible GTPases
Authors: Brown Hailey M. , Biering Scott B. , Zhu Allen , Choi Jayoung , Hwang Seungmin
Published: March 2018

A hallmark of positive‐sense RNA viruses is the formation of membranous shelters for safe replication in the cytoplasm. Once considered invisible to the immune system, these viral shelters are now found to be antagonized through the cooperation of autophagy proteins and anti‐microbial GTPases. This coordinated effort of autophagy proteins guiding GTPases functions against not only the shelters of viruses but also cytoplasmic vacuoles containing bacteria or protozoa, suggesting a broad immune‐defense mechanism against disparate vacuolar pathogens. Fundamental questions regarding this process remain: how the host recognizes these membranous structures as a target, how the autophagy proteins bring the GTPases to the shelters, and how the recruited GTPases disrupt these shelters. In this review, these questions are discussed, the answers to which will significantly advance our understanding of the response to vacuole‐like structures of pathogens, thereby paving the way for the development of broadly effective anti‐microbial strategies for public health.
New Roles for the Nucleolus in Health and Disease
Authors: Núñez Villacís Lorena , Wong Mei S. , Ferguson Laura L. , Hein Nadine , George Amee J. , Hannan Katherine M.
Published: March 2018

Over the last decade, our appreciation of the importance of the nucleolus for cellular function has progressed from the ordinary to the extraordinary. We no longer think of the nucleolus as simply the site of ribosome production, or a dynamic subnuclear body noted by pathologists for its changes in size and shape with malignancy. Instead, the nucleolus has emerged as a key controller of many cellular processes that are fundamental to normal cell homeostasis and the target for dysregulation in many human diseases; in some cases, independent of its functions in ribosome biogenesis. These extra‐nucleolar or new functions, which we term “non‐canonical” to distinguish them from the more traditional role of the nucleolus in ribosome synthesis, are the focus of this review. In particular, we explore how these non‐canonical functions may provide novel insights into human disease and in some cases new targets for therapeutic development.
Hawaiian Drosophila as an Evolutionary Model Clade: Days of Future Past
Authors: O'Grady Patrick , DeSalle Rob
Published: March 2018

The Hawaiian Drosophila have been a model system for evolutionary, ecological, and ethological studies since the inception of the Hawaiian Drosophila Project in the 1960s. Here we review the past and present research on this incredible lineage and provide a prospectus for future directions on genomics and microbial interactions. While the number of publications on this group has waxed and waned over the years, we assert that recent systematic, biogeographic, and ecological studies have reinvigorated Hawaiian Drosophila as an evolutionary model system. The characteristics that distinguish good model clades from good model organisms (e.g., Drosophila melanogaster) are somewhat different so we first define what constitutes a good evolutionary model. We argue that the Hawaiian Drosophila possess many desired aspects of a good evolutionary model, describe how this group of geographically isolated flies have been used in the past, and propose some exciting avenues for future evolutionary research on this diverse, dynamic clade of Drosophila.
How Does a Helicase Unwind DNA? Insights from RecBCD Helicase
Authors: Lohman Timothy M. , Fazio Nicole T.
Published: March 2018

DNA helicases are a class of molecular motors that catalyze processive unwinding of double stranded DNA. In spite of much study, we know relatively little about the mechanisms by which these enzymes carry out the function for which they are named. Most current views are based on inferences from crystal structures. A prominent view is that the canonical ATPase motor exerts a force on the ssDNA resulting in “pulling” the duplex across a “pin” or “wedge” in the enzyme leading to a mechanical separation of the two DNA strands. In such models, DNA base pair separation is tightly coupled to ssDNA translocation of the motors. However, recent studies of the Escherichia coli RecBCD helicase suggest an alternative model in which DNA base pair melting and ssDNA translocation occur separately. In this view, the enzyme‐DNA binding free energy is used to melt multiple DNA base pairs in an ATP‐independent manner, followed by ATP‐dependent translocation of the canonical motors along the newly formed ssDNA tracks. Repetition of these two steps results in processive DNA unwinding. We summarize recent evidence suggesting this mechanism for RecBCD helicase action.
On the performance of adaptive preprocessing technique in analyzing high‐dimensional censored data
Authors: Khan Md Hasinur Rahaman
Published: March 2018

Preprocessing for high‐dimensional censored datasets, such as the microarray data, is generally considered as an important technique to gain further stability by reducing potential noise from the data. When variable selection including inference is carried out with high‐dimensional censored data the objective is to obtain a smaller subset of variables and then perform the inferential analysis using model estimates based on the selected subset of variables. This two stage inferential analysis is prone to circularity bias because of the noise that might still remain in the dataset. In this work, I propose an adaptive preprocessing technique that uses sure independence screening (SIS) idea to accomplish variable selection and reduces the circularity bias by some popularly known refined high‐dimensional methods such as the elastic net, adaptive elastic net, weighted elastic net, elastic net‐AFT, and two greedy variable selection methods known as TCS, PC‐simple all implemented with the accelerated lifetime models. The proposed technique addresses several features including the issue of collinearity between important and some unimportant covariates, which is often the case in high‐dimensional setting under variable selection framework, and different level of censoring. Simulation studies along with an empirical analysis with a real microarray data, mantle cell lymphoma, is carried out to demonstrate the performance of the adaptive pre‐processing technique.
The use of a multiple schedule to decrease toe walking in a child with autism
Authors: Hodges Ansley C. , Wilder David A. , Ertel Hallie
Published: March 2018

We evaluated a procedure in which a wristband worn by a young boy with autism signaled the availability of praise for appropriate walking and the delivery of reprimands for toe walking. In the absence of the wristband, no programmed consequences were delivered for appropriate walking or toe walking. Once toe walking decreased to criterion levels with the wristband on, we increased the number of steps during which the participant wore the wristband. Results suggest that the procedure was effective to decrease toe walking in both clinic and community settings. To assess generalization, we had the participant's mother implement the procedure; it remained effective when implemented by her.
Mechanisms driving the lactate switch in Chinese hamster ovary cells
Authors: Hartley Fiona , Walker Tracy , Chung Vicky , Morten Karl
Published: March 2018

The metabolism of Chinese Hamster Ovary (CHO) cells in a production environment has been extensively investigated. However, a key metabolic transition, the switch from lactate production to lactate consumption, remains enigmatic. Though commonly observed in CHO cultures, the mechanism(s) by which this metabolic shift is triggered is unknown. Despite this, efforts to control the switch have emerged due to the association of lactate consumption with improved cell growth and productivity. This review aims to consolidate current theories surrounding the lactate switch. The influence of pH, NAD+/NADH, pyruvate availability and mitochondrial function on lactate consumption are explored. A hypothesis based on the cellular redox state is put forward to explain the onset of lactate consumption. Various techniques implemented to control the lactate switch, including manipulation of the culture environment, genetic engineering, and cell line selection are also discussed.
Unexpected instabilities explain batch‐to‐batch variability in cell‐free protein expression systems
Authors: Hunter Dominic J.B. , Bhumkar Akshay , Giles Nichole , Sierecki Emma , Gambin Yann
Published: March 2018

Cell‐free methods of protein synthesis offer rapid access to expressed proteins. Though the amounts produced are generally only at a small scale, these are sufficient to perform protein‐protein interaction assays and tests of enzymatic activity. As such they are valuable tools for the biochemistry and bioengineering community. However the most complex, eukaryotic cell‐free systems are difficult to manufacture in house and can be prohibitively expensive to obtain from commercial sources. The Leishmania tarentolae system offers a relatively cheap alternative which is capable of producing difficult to express proteins, but which is simpler to produce in large scale. However, this system suffers from batch‐to‐batch variability, which has been accepted as a consequence of the complexity of the extracts. Here we show an unexpected origin for the variability observed and demonstrate that small variations in a single parameter can dramatically affect expression, such that minor pipetting errors can have major effects on yields. L. tarentolae cell‐free lysate activity is shown to be more stable to changes in Mg2+ concentration at a lower ratio of feed solution to lysate in the reaction than typically used, and a higher Mg2+ optimum. These changes essentially eliminate batch‐to‐batch variability of L. tarentolae lysate activity and permit their full potential to be realized.
Population-based cohort study of surgical myotomy and pneumatic dilatation as primary interventions for oesophageal achalasia
Authors: Markar S R , Mackenzie H , Askari A , Faiz O , Hoare J , Zaninotto G , Hanna G B
Published: March 2018

Background

The aim of this national population-based cohort study was to compare rates of reintervention after surgical myotomy versus sequential pneumatic dilatation for the primary management of oesophageal achalasia.

Methods

Patients with oesophageal achalasia diagnosed between 2002 and 2012, and without an intervention in the preceding 5 years were identified from the Hospital Episode Statistics database. Patients were divided into two groups based on the primary treatment, and propensity score matching was used to compensate for differences in baseline characteristics.

Results

Some 14 705 patients were diagnosed with oesophageal achalasia, of whom 7487 (50·9 per cent) received interventional treatment: 1742 (23·3 per cent) surgical myotomy, 4534 (60·6 per cent) pneumatic dilatation and 1211 (16·2 per cent) endoscopic botulinum toxin injection. As age increased, the proportion of patients receiving myotomy decreased and the proportion undergoing dilatation increased. Patients who underwent surgical myotomy were younger (mean age 44·8 years versus 58·5 years among those who had pneumatic dilatation; P < 0·001), a greater proportion had a Charlson co-morbidity index score of 0 (90·1 versus 87·7 per cent; P = 0·003) and they were more commonly men (55·6 versus 51·8 per cent; P = 0·020). Following propensity score matching, the safety of the two initial treatment approaches was equivalent, with no difference in incidence of oesophageal perforation (1·3 and 1·4 per cent after myotomy and dilatation respectively; P = 0·750). However, dilatation was associated with increased need for reintervention (59·6 versus 13·8 per cent; P < 0·001) and frequency of reinterventions (mean 0·34 versus 0·06 per year; P < 0·001).

Conclusion

Surgical myotomy was associated with a lower rate of reintervention and could be offered as primary treatment in patients with oesophageal achalasia who are fit for surgery. For those unfit for surgery, pneumatic dilatation may provide symptomatic relief with approximately 60 per cent of patients requiring reintervention.
Thirty-day mortality in patients undergoing laparotomy for small bowel obstruction
Authors: Peacock O , Bassett M G , Kuryba A , Walker K , Davies E , Anderson I , Vohra R S
Published: March 2018

Background

Small bowel obstruction (SBO) is a common indication for emergency laparotomy. There are currently variations in the timing of surgery for patients with SBO and limited evidence on whether delayed surgery affects outcomes. The aim of this study was to evaluate the impact of time to operation on 30-day mortality in patients requiring emergency laparotomy for SBO.

Methods

Data were collected from the National Emergency Laparotomy Audit (NELA) on all patients aged 18 years or older who underwent emergency laparotomy for all forms of SBO between December 2013 and November 2015. The primary outcome measure was 30-day mortality, with date of death obtained from the Office for National Statistics. Patients were grouped according to the time from admission to surgery (less than 24 h, 24–72 h and more than 72 h). A multilevel logistic regression model was used to explore the impact of patient factors, primarily delay to surgery, on 30-day mortality.

Results

Some 9991 patients underwent emergency laparotomy requiring adhesiolysis or small bowel resection for SBO. The overall mortality rate was 7·2 per cent (722 patients). Within each time group, 30-day mortality rates were significantly worse with increasing age, ASA grade, Portsmouth POSSUM score and level of contamination. Patients undergoing emergency laparotomy more than 72 h after admission had a significantly higher risk-adjusted 30-day mortality rate (odds ratio 1·39, 95 per cent c.i. 1·09 to 1·76).

Conclusion

In patients who require an emergency laparotomy with adhesiolysis or resection for SBO, a delay to surgery of more than 72 h is associated with a higher 30-day postoperative mortality rate.
Associations between apparent diffusion coefficient and electromyography parameters in myositis—A preliminary study
Authors: Meyer Hans‐Jonas , Emmer Alexander , Kornhuber Malte , Surov Alexey
Published: March 2018

Objective

MRI is widely used in several muscle disorders. Diffusion‐weighted imaging (DWI) is an emergent imaging modality sensitive to microstructural alterations in tissue. The apparent diffusion coefficient (ADC) is used to quantify the random motion of water molecules. Electromyography (EMG) is a clinically used diagnostic tool in myositis. The aim of this study was to elucidate possible associations between ADC values and EMG findings in myositis patients.

Method

Seven patients (eight investigated muscles) with myositis (mean age 51.43 ± 19 years) were included in this study. The diagnosis was confirmed by histopathology in every case. DWI was obtained with a 1.5‐T scanner using two b‐values 0 and 1000 s/mm². In all patients, a needle electromyography (EMG) was performed within 3 days to the MRI. The following EMG parameters were studied: motor unit action potential (MUAP) amplitudes and durations, as well as pathological spontaneous activity. Spearman's correlation coefficient was used to analyze associations between investigated parameters.

Results

The estimated mean ADCmean value was 1.51 ± 0.29 × 10−3 mm²/s, mean ADCmin was 1.28 ± 0.27 × 10−3 mm²/s, and mean ADCmax was 1.73 ± 0.28 × 10−3 mm²/s. Correlation analysis identified significant associations between ADCmean and duration of the MUAP (p = .78 P = .0279) and between ADCmin and duration of the MUAP (p = .85, P = .01). There were no significant differences according to pathological spontaneous activity.

Conclusion

ADCmean and ADCmin showed strong positive correlations with the duration of the MUAP in myositis patients. Both modalities might similarly reflect muscle fiber loss in myositis patients.
An overview of protective strategies against ischemia/reperfusion injury: The role of hyperbaric oxygen preconditioning
Authors: Hentia Ciprian , Rizzato Alex , Camporesi Enrico , Yang Zhongjin , Muntean Danina M. , Săndesc Dorel , Bosco Gerardo
Published: March 2018

Introduction

Ischemia/reperfusion (I/R) injury, such as myocardial infarction, stroke, and peripheral vascular disease, has been recognized as the most frequent causes of devastating disorders and death currently. Protective effect of various preconditioning stimuli, including hyperbaric oxygen (HBO), has been proposed in the management of I/R.

Methods

In this study, we searched and reviewed up‐to‐date published papers to explore the pathophysiology of I/R injury and to understand the mechanisms underlying the protective effect of HBO as conditioning strategy.

Results

Animal study and clinic observation support the notion that HBO therapy and conditioning provide beneficial effect against the deleterious effects of postischemic reperfusion. Several explanations have been proposed. The first likely mechanism may be that HBO counteracts hypoxia and reduces I/R injury by improving oxygen delivery to an area with diminished blood flow. Secondly, by reducing hypoxia–ischemia, HBO reduces all the pathological events as a consequence of hypoxia, including tissue edema, increased affective area permeability, postischemia derangement of tissue metabolism, and inflammation. Thirdly, HBO may directly affect cell apoptosis, signal transduction, and gene expression in those that are sensitive to oxygen or hypoxia. HBO provides a reservoir of oxygen at cellular level not only carried by blood, but also by diffusion from the interstitial tissue where it reaches high concentration that may last for several hours, improves endothelial function and rheology, and decreases local inflammation and edema.

Conclusion

Evidence suggests the benefits of HBO when used as a preconditioning stimulus in the setting of I/R injury. Translating the beneficial effects of HBO into current practice requires, as for the “conditioning strategies”, a thorough consideration of risk factors, comorbidities, and comedications that could interfere with HBO‐related protection.
Eleven years’ experience with Intrathecal Baclofen – Complications, risk factors
Authors: Pucks‐Faes Elke , Hitzenberger Gabriel , Matzak Heinrich , Fava Elena , Verrienti Giulio , Laimer Ilse , Fritz Josef , Saltuari Leopold
Published: March 2018

Objective

Treatment with intrathecal baclofen (ITB) is commonly used in patients with severe spasticity. However, complications may occur after implantation of the ITB‐device, albeit mainly procedure‐ and device‐related problems. The aim of the study was to assess surgical‐ as well as catheter‐ and pump‐related complications and define their risk factors.

Methods

We retrospectively evaluated all patients with an implanted ITB‐device who were treated at the Department of Neurology, Hochzirl Hospital, Zirl, Austria, between 2006 and 2016.

Results

Twenty‐nine of 116 (25%) patients experienced 32 complications: 5 procedure‐ and 27 device‐related (4 pump‐ and 23 catheter‐associated) problems occurred. Risk factors for sustaining any complication were a spinal localization of lesion (odds ratio [OR] OR 2.71, p = .021), other catheter types than an Ascenda® catheter (OR 3.87, p = .041), a lower modified Rankin Scale (median 4 vs. 5; OR 2.86, p = .015) and a higher Barthel Index (median 53 vs. 0; OR 2.84, p = .006). The median time from the last ITB‐related surgery to the first complication was 18 (IQR 1‐57) months. Overall, 47% complications occurred within the first year after any surgical procedure regarding the ITB‐device, thereof 25% within the first month.

Conclusions

Procedure‐ and device‐related complications are frequent after implantation of an ITB‐device with catheter‐associated complications as the most frequently encountered problems. Patients with a spinal origin of spasticity, a lower modified Rankin Scale and a higher Barthel Index have a higher risk to sustain a complication.
In vitro studies on the degradation of poly(cis‐1,4‐isoprene)
Authors: Andler R. , Altenhoff A.‐L. , Mäsing F. , Steinbüchel A.
Published: March 2018

Cleavage of the backbone of poly(cis‐1,4‐isoprene) (IR) in solid rubber material was accomplished by the addition of partially purified latex clearing protein (Lcp1VH2) using a 200‐mL enzyme reactor. Two strategies for the addition of Lcp1VH2 were studied revealing that the daily addition of 50 µg mL−1 of Lcp1VH2 for 5 days was clearly a more efficient regime in comparison to a one‐time addition of 250 µg of Lcp1VH2 at the beginning. Soluble oligo(cis‐1,4‐isoprene) molecules occurred as degradation products and were identified by ESI‐MS and GPC. Oxygenase activity of Lcp1VH2 with solid IR particles as substrate was shown for the first time by measuring the oxygen consumption in the reaction medium. A strong decrease of the dissolved oxygen concentration was detected at the end of the assay, which indicates an increase in the number of cleavage reactions. The oligo(cis‐1,4‐isoprene) molecules comprised 1 to 11 isoprene units and exhibited an average molecular weight (Mn) of 885 g mol−1. Isolation of the oligo(cis‐1,4‐isoprene) molecules was achieved by using silica gel column chromatography. The relative quantification of the isolated products was performed by HPLC‐MS after derivatization with 2,4‐dinitrophenilhydrazyne yielding a concentration of total degradation products of 1.62 g L−1. Analysis of the polymer surface in samples incubated for 3 days with Lcp1VH2 via ATR‐FTIR indicated the presence of carbonyl groups, which occurred upon the cleavage reaction. This study presents a cell‐free bioprocess as an alternative rubber treatment that can be applied for the partial degradation of the polymer. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:890–899, 2018
Insulin secretion kinetics from single islets reveals distinct subpopulations
Authors: Guruswamy Damodaran Rajesh , Poussard Alexandre , Côté Benoît , Andersen Parker L. , Vermette Patrick
Published: March 2018

Type II diabetes progresses with inadequate insulin secretion and prolonged elevated circulating glucose levels. Also, pancreatic islets isolated for transplantation or tissue engineering can be exposed to glucose over extended timeframe. We hypothesized that isolated pancreatic islets can secrete insulin over a prolonged period of time when incubated in glucose solution and that not all islets release insulin in unison. Insulin secretion kinetics was examined and modeled from single mouse islets in response to chronic glucose exposure (2.8‐20 mM). Results with single islets were compared to those from pools of islets. Kinetic analysis of 58 single islets over 72 h in response to elevated glucose revealed distinct insulin secretion profiles: slow‐, fast‐, and constant‐rate secretors, with slow‐secretors being most prominent (ca., 50%). Variations in the temporal response to glucose therefore exist. During short‐term (<4 h) exposure to elevated glucose few islets are responding with sustained insulin release. The model allowed studying the influence of islet size, revealing no clear effect. At high‐glucose concentrations, when secretion is normalized to islet volume, the tendency is that smaller islets secrete more insulin. At high‐glucose concentrations, insulin secretion from single islets is representative of islet populations, while under low‐glucose conditions pooled islets did not behave as single ones. The characterization of insulin secretion over prolonged periods complements studies on insulin secretion performed over short timeframe. Further investigation of these differences in secretion profiles may resolve open‐ended questions on pre‐diabetic conditions and transplanted islets performance. This study deliberates the importance of size of islets in insulin secretion. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1059–1068, 2018
Development of generic raman models for a GS‐KOTM CHO platform process
Authors: Webster Thaddaeus A. , Hadley Brian C. , Hilliard William , Jaques Colin , Mason Carrie
Published: March 2018

The monitoring and control of bioprocesses is of the utmost importance in order to provide a consistent, safe, and high‐quality product for consumers. Current monitoring and control schemes rely on infrequent and time consuming offline sampling methods, which inherently leads to some variability in the process which may impact the product quality profile. As part of Lonza's dedication to process analytical technology (PAT) initiatives this study evaluated the ability to generate generic calibration models, which are independent of the cell line, using Raman probes to monitor changes in glucose, lactate, glutamate, ammonium, viable cell concentration (VCC), total cell concentration (TCC) and product concentration. Calibration models were developed from cell culture using two different CHOK1SV GS‐KOTM cell lines producing different monoclonal antibodies (mAbs). Developed predictive models, measured changes in glucose, lactate, ammonium, VCC, and TCC with average prediction errors of 0.44, 0.23, 0.03 g L−1, 1.90 × 106 cells mL−1, and 1.85 × 106 cells mL−1, respectively over the course of cell culture with minimal cell line dependence. The development of these generic models allows the application of spectroscopic PAT techniques in clinical and commercial manufacturing environments, where processes are typically run once or twice in GMP manufacturing based on a common platform process. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:730–737, 2018
Behavior of several pseudomonas putida strains growth under different agitation and oxygen supply conditions
Authors: Rodriguez Alberto , Escobar Sebastian , Gomez Emilio , Santos Victoria E. , Garcia‐Ochoa Felix
Published: March 2018

The growth rate of four strains of Pseudomonas putida, KT2440, KT2442, KTH2, and KTH2 (pESOX3), under different fluid dynamic conditions has been studied. The cultures were conducted in a stirred tank bioreactor by changing the stirrer speed. Several process variables, such as biomass concentration, dissolved oxygen concentration, oxygen mass transfer rate and oxygen uptake rate, have been measured or calculated. Also cell viability was determined by viable colony counting in Petri dishes and culture samples were subjected into a transmission electron microscopy analysis, in order to describe the integrity of the individual cells. The experimental results show that the genetically modified organisms, the strains KTH2 and KTH2 (pESOX3), present a different growth under low agitation conditions, and low oxygen supply level, while the growth of the wild type strains, KT2440 and KT2442, followed the typical sigmoidal evolution that could be described by the logistic equation. The presence of outer membrane vesicles has been observed in the GMO strains. When the cultures were conducted at low stirrer speed, and so at low oxygen transfer rate, these vesicles were detected, indicating the bacterial response to oxidative stress, caused by the catalytic activity of the HpaC enzyme. For all of the strains tested, no hydrodynamic stress has been detected, even at very high agitation levels. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:900–909, 2018
Monitoring mAb cultivations with in‐situ raman spectroscopy: The influence of spectral selectivity on calibration models and industrial use as reliable PAT tool
Authors: Santos Rafael M. , Kessler Jean‐Michel , Salou Patrick , Menezes Jose C. , Peinado Antonio
Published: March 2018

Raman spectroscopy is a suitable monitoring technique for CHO cultivations. However, a thorough discussion of peaks, bands, and region assignments to key metabolites and culture attributes, and the interpretability of produced calibrations is scarce. That understanding is vital for the long‐term predictive ability of monitoring models, and to facilitate lifecycle management that comply with regulatory guidelines. Several fed‐batch lab‐scale mAb mammalian cultivations were carried out, with in situ Raman spectroscopy used for process state estimation and attribute monitoring. The goal was to evaluate its use as a process analytical technology (PAT) tool to detect residual glucose and lactate levels, understand their dynamics and interconversion, and eventually estimate key performance culture and product quality attributes. Glucose and lactate models were optimized up to 0.31 g L−1 with 3 Latent Variables (LVs) and 0.19 g L−1 (2 LVs) accuracy, respectively. Glutamine and product titer models, were not specific and accurate enough, even though indirect calibrations were obtained with a RMSEP of 0.12 g L−1 (4 LVs) and 0.29 g L−1 (5 LVs), respectively. A critical discussion and details about the extensive work done in calibration development and optimization are provided. Namely, considering a risk‐based selection of variability sources impacting sample spectra, executing designed experiments with spiked cultivations, and using advanced chemometric procedures for variable selection and model cross validation. A strategy is presented to evaluation Raman spectroscopy as a reliable PAT technology fit‐for industrial use. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:659–670, 2018
Trainee estimates of working alliance with upper‐ and working‐class clients
Authors: Kaiser Dakota J. , Prieto Loreto R.
Published: March 2018

Background

Although social class is recognized as an important variable within multicultural psychology, relatively little research has been conducted on the effects of social class on psychotherapy processes and outcomes.

Aims

Using a vignette‐based methodology, we examined the effects of therapy trainees’ social class identity and attitudes on their estimation of forming a working alliance with clients from an upper‐ and working‐class status.

Materials & Methods

Participants were randomly assigned to read one of two clinical vignettes of either an upper‐ or working‐class client presenting for individual therapy. Participants then rated the anticipated strength of their working alliance with the client and their attitudes related to social class.

Results

Regarding training, our results indicated that participants received very few clock hours of instruction related to social class issues. After controlling for socially desirable responding, although we detected no statistically significant bias against lower‐class clientele, we did find a measure of bias among trainees towards those from the upper class. Lower trainee social class; a greater belief that social class status is earned through personal actions, merit and ability; and a greater level of anticipated difficulty working with their vignette client explained the greatest amount of trainee bias towards those from the upper social class.

Discussion

We discussed the implications of these findings for training and future research.
The rosy side and the blue side of emotional brand attachment
Authors: Hung Hsiu‐Yu , Lu Hao‐Te
Published: March 2018

This paper examines the rosy side (positive emotion) and the blue side (negative emotion) of brand attachment in consumer–brand relationships and their distinct motivational triggers. Adopting attachment theory, self‐determination theory, and the relationship investment model, this paper provides a richer understanding of the positivity and negativity of emotional brand attachment in terms of consequential branding outcomes. Two hierarchical constructs—autonomous and controlled motivations—are proposed and examined using partial least squares path modelling. Data were collected through a 3‐stage survey to gain 507 samples of customers' brand attachments to 9 international car brands. This research reveals that customers' positive‐side (rosy‐side) emotional brand attachments are driven by autonomous motivations including satisfying the intrinsic motivation of pursuing pleasure and fulfilling needs for self‐determination and self‐congruence in the car brand consumption context. Customers' negative‐side (blue‐side) emotional brand attachment reflects feelings of separation distress, anxiety, and sadness that are triggered by their controlled motivations in car brand consumption contexts. The results indicate that positive emotions (the rosy side) of brand attachment are a more effective predictor of brand repurchasing intentions and word‐of‐mouth behaviours than are negative emotions (the blue side) of brand attachment. This study applies Deci and Ryan's autonomous and controlled motivations to explain the positivity and negativity of consumers' emotional brand attachment in brand–consumer relationships.
Directed Evolution of Hyaluronic Acid Synthase from Pasteurella multocida towards High‐Molecular‐Weight Hyaluronic Acid
Authors: Mandawe John , Infanzon Belen , Eisele Anna , Zaun Henning , Kuballa Jürgen , Davari Mehdi D. , Jakob Felix , Elling Lothar , Schwaneberg Ulrich
Published: March 2018

Hyaluronic acid (HA), with diverse cosmetic and medical applications, is the natural glycosaminoglycan product of HA synthases. Although process and/or metabolic engineering are used for industrial HA production, the potential of protein engineering has barely been realised. Herein, knowledge‐gaining directed evolution (KnowVolution) was employed to generate an HA synthase variant from Pasteurella multocida (pmHAS) with improved chain‐length specificity and a twofold increase in mass‐based turnover number. Seven improved pmHAS variants out of 1392 generated by error‐prone PCR were identified; eight prospective positions were saturated and the most beneficial amino acid substitutions were recombined. After one round of KnowVolution, the longest HA polymer (<4.7 MDa), through an engineered pmHAS variant in a cell‐free system, was synthesised. Computational studies showed that substitutions from the best variant (T40L, V59M and T104A) are distant from the glycosyltransferase sites and increase the flexibility of the N‐terminal region of pmHAS. Taken together, these findings suggest that the N terminus may be involved in HA synthesis and demonstrate the potential of protein engineering towards improved HA synthase activity.
Protein Crystallography and Site‐Direct Mutagenesis Analysis of the Poly(ethylene terephthalate) Hydrolase PETase from Ideonella sakaiensis
Authors: Liu Bing , He Lihui , Wang Liping , Li Tao , Li Changcheng , Liu Huayi , Luo Yunzi , Bao Rui
Published: March 2018

Unlike traditional recycling strategies, biodegradation is a sustainable solution for disposing of poly(ethylene terephthalate) (PET) waste. PETase, a newly identified enzyme from Ideonella sakaiensis, has high efficiency and specificity towards PET and is, thus, a prominent candidate for PET degradation. On the basis of biochemical analysis, we propose that a wide substrate‐binding pocket is critical for its excellent ability to hydrolyze crystallized PET. Structure‐guided site‐directed mutagenesis revealed an improvement in PETase catalytic efficiency, providing valuable insight into how the molecular engineering of PETase can optimize its application in biocatalysis.
Biochemical Characterization of β‐Amino Acid Incorporation in Fluvirucin B2 Biosynthesis
Authors: Barajas Jesus F. , Zargar Amin , Pang Bo , Benites Veronica T. , Gin Jennifer , Baidoo Edward E. K. , Petzold Christopher J. , Hillson Nathan J. , Keasling Jay D.
Published: March 2018

Naturally occurring lactams, such as the polyketide‐derived macrolactams, provide a diverse class of natural products that could enhance existing chemically produced lactams. Although β‐amino acid loading in the fluvirucin B2 polyketide pathway was proposed by a previously identified putative biosynthetic gene cluster, biochemical characterization of the complete loading enzymes has not been described. Here we elucidate the complete biosynthetic pathway of the β‐amino acid loading pathway in fluvirucin B2 biosynthesis. We demonstrate the promiscuity of the loading pathway to utilize a range of amino acids and further illustrate the ability to introduce non‐native acyl transferases to selectively transfer β‐amino acids onto a polyketide synthase (PKS) loading platform. The results presented here provide a detailed biochemical description of β‐amino acid selection and will further aid in future efforts to develop engineered lactam‐producing PKS platforms.
Steroidal hormone and morphological responses in the prostate anterior lobe in different cancer grades after Celecoxib and Goniothalamin treatments in TRAMP mice
Authors: Silva Rafael Sauce , Kido Larissa Akemi , Montico Fabio , Vendramini‐Costa Débora Barbosa , Pilli Ronaldo Aloise , Cagnon Valeria Helena Alves
Published: March 2018

Prostate cancer is the second most diagnosed cancer in the world, and alternative methods to prevent and treat different lesion grades need to be evaluated. The objective was to evaluate the morphological, hormonal, and inflammatory responses in the prostate anterior lobe in transgenic adenocarcinoma of the mouse prostate (TRAMP), following Celecoxib and Goniothalamin (GTN) treatments. All animals were treated for 4 weeks, from 8 weeks of age and euthanized either immediately after treatment (12‐week‐old mice: immediate response) or later (22‐week‐old mice: late response). The results showed a significant increase of high‐grade prostatic intraepithelial neoplasia (HGPIN) and well‐differentiated adenocarcinoma (WDA), according to the age in the control groups. Celecoxib treatment decreased the WDA incidence in the late response group. GTN led to a significant healthy tissue increase, and an LGPIN and HGPIN decrease in the immediate response group. In the late response group, GTN led to healthy area increase and there was no occurrence of WDA. AR and ERα immunoexpressions were reduced by both treatments in the immediate response groups. However, only GTN was able to decrease the ERα level in the late response group. Regarding COX‐2 immunoreactivity, both treatments reduced the frequency of this enzyme. We can conclude that the prostate anterior lobe is a good model to study prostate cancer, considering its slow progression. Both treatments led to cancer delay in the prostate anterior lobe. However, GTN pointed towards a better treatment spectrum in the signaling pathways in the prostate microenvironment, particularly in ERα.
Falling number of soft white wheat by near‐infrared spectroscopy: A challenge revisited
Authors: Delwiche Stephen R. , Higginbotham Ryan W. , Steber Camille M.
Published: March 2018

Background and objectives

Wheat Hagberg falling number (FN) is a long‐standing quality test that, by means of measuring the viscosity of a heated water‐meal or water‐flour mixture, characterizes the activity of endogenous α‐amylase, the enzyme primarily responsible for starch hydrolysis. The accuracy, time requirement, and cost of this test have come under heightened scrutiny, particularly in seasons when weather conditions have been favorable to preharvest sprouting or late maturity amylase. Near‐infrared (NIR) spectroscopy, an analytical approach routinely used in the grain industry to measure contents of protein and moisture, was reexamined as a possible alternative to the FN procedure.

Findings

Partial least squares (PLS) regression quantitative models developed on a genetically diverse set of Washington grown white wheat demonstrated low accuracy, with standard errors of performance ranging from 40 to 77 s. Alternatively, linear discriminant analysis and PLS discriminant analysis (PLSDA) qualitative models, developed and tested using a FN cutoff (pass/fail) value, also demonstrated low accuracy, with the best model correctly identifying 67% and 71% of the samples, respectively, above and below a threshold value established as the median value of FN in a calibration set of several hundred samples.

Conclusions

Replacement of the FN test with one based on NIR spectroscopy on either whole grain or ground meal for making decisions on segregating wheat lots according to α‐amylase activity is not recommended.

Significance and novelty

Because NIR spectroscopy is not sufficiently accurate to quantitatively model FN or differentiate low from high FN grain, viscometry procedures for starch integrity, such as FN, will continue their use in grain commerce.
Effect of organic acids and NaCl on the rheological properties of dough prepared using Pembina and Harvest CWRS wheat cultivars
Authors: Stone Andrea K. , Lam Ricky S. H. , Hopkins Erin J. , Hucl Pierre , Scanlon Martin G. , Nickerson Michael T.
Published: March 2018

Background and objectives

The effect of organic acids (1.2 mmol/100 g flour: acetic, citric, lactic or fumaric acid) and NaCl content (0%, 1% and 2%, flour weight basis) on the rheological properties and stickiness of dough prepared using two CWRS wheat cultivars (Harvest and Pembina) was investigated as a means of understanding the interaction of salt and acid on dough viscoelastic behavior.

Findings

Overall higher salt levels made the doughs stiffer and less sticky as evident by |G*| and Jel increasing and dough stickiness decreasing as salt content increased. The addition of citric, acetic, and lactic acids caused an overall increase in |G*| and Jel, and a decrease in tan δ and Jmax, whereas fumaric acid had the inverse effect. Dough stickiness increased regardless of the type of acid added at all salt levels but more so when no salt was added to the dough. The effect of acid addition on dough rheology was cultivar dependent as evident by a small overall increase in |G*| with acid addition to Pembina and an overall decrease in |G*| when using Harvest.

Conclusions

The effect of cultivar, salt content and acid type was highly interdependent for the dough complex modulus, tan δ, maximum creep compliance, and relative elasticity along with dough stickiness.

Significance and novelty

This research provides fundamental knowledge on the effect of salt reduction and acidulants on dough rheology with a view to understanding how water incorporation in the gluten network affects attempts to reduce sodium content in the dough without impairing dough handling properties.
Focal atrophy of the unilateral masticatory muscles caused by pure trigeminal motor neuropathy: case report
Authors: Kämppi Antti , Kämppi Leena , Kemppainen Pentti , Kanerva Mari , Toppila Jussi , Auranen Mari
Published: March 2018

Key Clinical Message

Patients with unknown clinical or radiological asymmetry in the face structures combined with atrophy and weakness of the masticatory muscles should be comprehensively examined clinically and with MRI, neurophysiological measurements, and serologically. Malignant lesions or benign idiopathic unilateral trigeminal motor neuropathy should be considered as an etiological explanation for the asymmetry.
DOPA and Adrenaline Oxidation Kinetics and Intermediates Identified by Electrospray Ionization Mass Spectrometry in Real Time
Authors: Iftikhar Imran , El‐Nour Kholoud Mohammed Abou , Brajter‐Toth Anna
Published: March 2018

DOPA and adrenaline (Adr) oxidation intermediates were identified using electrospray ionization mass spectrometry (ESI MS) by exploiting electrochemical oxidation reactions that occur during ESI. In addition, on‐line MS monitoring of the oxidations allowed evaluation of DOPA and Adr oxidation kinetics with approximately 3 ms time resolution. Kinetic information was confirmed by on‐line electrochemistry/ESI MS (EC/ESI MS). Application of an EC cell voltage in EC/ESI MS increased the oxidation rates and MS detection sensitivity of intermediates. During ESI and EC/ESI MS, oxidation rates were
, cyclization rates of quinone oxidation products were
, and cyclic leucochrome oxidation rates were DA>Adr>DOPA. Formation of radical semiquinones of Adr and DOPA through OH. radical‐initiated oxidation pathways was indicated, as reported for DA. Images of catecholamine oxidation pathways obtained with high time resolution, using MS, underscore the differences between the reactivity of DOPA, Adr, and DA.