Biotechnology and BioengineeringVolume 116, Issue 7 p. 1563-1566 ISSUE INFORMATIONFree Access Biotechnology and Bioengineering: Volume 116, Number 7, July 2019 First published: 01 June 2019 https://doi.org/10.1002/bit.26761AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Volume116, Issue7July 2019Pages 1563-1566 RelatedInformation

The front cover image is based on the Article Adenovirus 5 recovery using nanofiber ion-exchange adsorbents by Jordan Turnbull et al., DOI: 10.1002/bit.26972.

Viral vectors such as adenovirus have successful applications in vaccines and gene therapy but the manufacture of the high-quality virus remains a challenge. It is desirable to use the adsorption-based chromatographic separations that so effectively underpin the therapeutic protein manufacture. However fundamental differences in the size and stability of this class of product mean it is necessary to revisit the design of sorbent's morphology and surface chemistry. In this study, the behaviour of a cellulose nanofiber ion-exchange sorbent derivatised with quaternary amine ligands at defined densities is characterised to address this. This material was selected as it has a large accessible surface area for viral particles and rapid process times. Initially, the impact of surface chemistry on infective product recovery using low (440 µmol/g), medium (750 µmol/g), and high (1029 µmol/g) ligand densities is studied. At higher densities product stability is reduced, this effect increased with prolonged adsorption durations of 24 min with just ~10% loss at low ligand density versus ~50% at high. This could be mitigated by using a high flow rate to reduce the cycle time to ~1 min. Next, the impact of ligand density on the separation's resolution was evaluated. Key to understanding virus quality is the virus particle: infectious virus particle ratio. It was found this parameter could be manipulated using ligand density and elution strategy. Together this provides a basis for viral vector separations that allows for their typically low titres and labile nature by using high liquid velocity to minimise both load and on-column times while separating key product and process-related impurities.

[This corrects the article DOI: 10.3389/fpsyg.2013.00088.].

Large scale continuous cell-line cultures promise greater reproducibility and efficacy for the production of influenza vaccines, and adenovirus for gene therapy. This paper seeks to use an existing validated ultra scale-down tool, which is designed to mimic the commercial scale process environment using only milliliters of material, to provide some initial insight into the performance of the harvest step for these processes. The performance of industrial scale centrifugation and subsequent downstream process units is significantly affected by shear. The properties of these cells, in particular their shear sensitivity, may be changed considerably by production of a viral product, but literature on this is limited to date. In addition, the scale-down tool used here has not previously been applied to the clarification of virus production processes. The results indicate that virus infected cells do not actually show any increase in sensitivity to shear, and may indeed become less shear sensitive, in a similar manner to that previously observed in old or dead cell cultures. Clarification may be most significantly dependent on the virus release mechanism, with the budding influenza virus producing a much greater decrease in clarification than the lytic, non-enveloped adenovirus. A good match was also demonstrated to the industrial scale performance in terms of clarification, protein release, and impurity profile.

Title: Ultra scale-down approaches to study the centrifugal harvest for viral vaccine production. † Short running title: Centrifugation ultra scale-down for viral vaccines Author List: Beatrice J Melinek, Sandrine Dessoy*, Bernice Wright, Dan G Bracewell, Tarit K Mukhopadhyay Affiliations: + Dept. of Biochemical Engineering, Bernard Katz building, University College London, Gordon Street, London, WC1H 0AH * GSK Vaccines, Rixensart, Belgium Email for correspondence: ucbetkm@ucl.ac.uk Author Telephone numbers: Beatrice J Melinek: 07984330623 Dan Bracewell: +44(0)2076799580 Tarit Mukhopadhyay: Extn. 30438

Despite growing evidence for demonstrated efficacy of mindfulness in various disorders, there is a continuous concern about the relationship between mindfulness practice and psychosis. As schizotypy is part of the psychosis spectrum, we examined the relationship between long-term mindfulness practice and schizotypy in two independent studies. Study 1 included 24 experienced mindfulness practitioners (19 males) from the Buddhist tradition (meditators) and 24 meditation-naïve individuals (all males). Study 2 consisted of 28 meditators and 28 meditation-naïve individuals (all males). All participants completed the Schizotypal Personality Questionnaire (Raine, 1991), a self-report scale containing 9 subscales (ideas of reference, excessive social anxiety, magical thinking, unusual perceptual experiences, odd/eccentric behavior, no close friends, odd speech, constricted affect, suspiciousness). Participants of study 2 also completed the Five-Facet Mindfulness Questionnaire which assesses observing (Observe), describing (Describe), acting with awareness (Awareness), non-judging of (Non-judgment) and non-reactivity to inner experience (Non-reactivity) facets of trait mindfulness. In both studies, meditators scored significantly lower on suspiciousness and higher on magical thinking compared to meditation-naïve individuals and showed a trend towards lower scores on excessive social anxiety. Excessive social anxiety correlated negatively with Awareness and Non-judgment; and suspiciousness with Awareness, Non-judgment and Non-reactivity facets across both groups. The two groups did not differ in their total schizotypy score. We conclude that mindfulness practice is not associated with an overall increase in schizotypal traits. Instead, the pattern suggests that mindfulness meditation, particularly with an emphasis on the Awareness, Non-judgment and Non-reactivity aspects, may help to reduce suspiciousness and excessive social anxiety.

Tissue engineering is a rapidly advancing field that is likely to transform how medicine is practised in the near future. For hollow organs such as those found in the cardiovascular and respiratory systems or gastrointestinal tract, tissue engineering can provide replacement of the entire organ or provide restoration of function to specific regions. Larger tissue-engineered constructs often require biomaterial-based scaffold structures to provide support and structure for new tissue growth. Consideration must be given to the choice of material and manufacturing process to ensure the de novo tissue closely matches the mechanical and physiological properties of the native tissue. This review will discuss some of the approaches taken to date for fabricating hollow organ scaffolds and the selection of appropriate biomaterials.

AIM:Fecal and urinary incontinence are devastating consequences of obstetric-related perineal injury. The aim of the present study is to determine acceptability to parous women of autologous cell-based therapy for fecal and urinary incontinence that arises due to pelvic diaphragm tearing during vaginal childbirth.MATERIALS & METHODS:A multiple choice questionnaire survey was offered to 76 parous women at the Maternity Unit, University College Hospital, London, UK. Seventy completed questionnaires - response rate: 92%.RESULTS:In total, 84% of women indicated a willingness to accept autologous cell-based therapy for obstetric injury-induced incontinence rather than surgery.CONCLUSION:These observational data provide an indication of likely acceptance of autologous cell-based therapies for birth injury incontinence and will help with designing new therapeutic approaches.

BACKGROUND:This study examined the effects of cultivated (i.e. developed through training) and dispositional (trait) mindfulness on smooth pursuit (SPEM) and antisaccade (AS) tasks known to engage the fronto-parietal network implicated in attentional and motion detection processes, and the fronto-striatal network implicated in cognitive control, respectively. METHODS:Sixty healthy men (19-59years), of whom 30 were experienced mindfulness practitioners and 30 meditation-naïve, underwent infrared oculographic assessment of SPEM and AS performance. Trait mindfulness was assessed using the self-report Five Facet Mindfulness Questionnaire (FFMQ). RESULTS:Meditators, relative to meditation-naïve individuals, made significantly fewer catch-up and anticipatory saccades during the SPEM task, and had significantly lower intra-individual variability in gain and spatial error during the AS task. No SPEM or AS measure correlated significantly with FFMQ scores in meditation-naïve individuals. CONCLUSIONS:Cultivated, but not dispositional, mindfulness is associated with improved attention and sensorimotor control as indexed by SPEM and AS tasks.

Cognitive impairment, particularly in attention, memory and executive function domains, is commonly present and associated with poor functional outcomes in schizophrenia. In healthy adults, environmental noise adversely affects many cognitive domains, including those known to be compromised in schizophrenia. This pilot study examined whether environmental noise causes further cognitive deterioration in a small sample of people with schizophrenia. Eighteen outpatients with schizophrenia on stable doses of antipsychotics and 18 age and sex-matched healthy participants were assessed on a comprehensive cognitive battery including measures of psychomotor speed, attention, executive functioning, working memory, and verbal learning and memory under three different conditions [quiet: ~30dB(A); urban noise: building site noise, 68–78dB(A); and social noise: background babble and footsteps from a crowded hall without any discernible words, 68–78dB(A)], 7–14days apart, with counter-balanced presentation of noise conditions across participants of both groups. The results showed widespread cognitive impairment in patients under all conditions, and noise-induced impairments of equal magnitude on specific cognitive functions in both groups. Both patient and healthy participant groups showed significant disruption of delayed verbal recall and recognition by urban and social noise, and of working memory by social noise, relative to the quiet condition. Performance under urban and social noise did not differ significantly from each other for any cognitive measure in either group. We conclude that noise has adverse effects on the verbal and working memory domains in schizophrenia patients and healthy participants. This may be particularly problematic for patients as it worsens their pre-existing cognitive deficits.

Poly (lactic-co-glycolic) acid microspheres are amenable to a number of biomedical procedures that support delivery of cells, drugs, peptides or genes. Hydrophilisation or wetting of poly (lactic-co-glycolic) acid are an important pre-requisites for attachment of cells and can be achieved via exposure to plasma oxygen or nitrogen, surface hydrolysis with NaOH or chloric acid, immersion in ethanol and water, or prolonged incubation in phosphate buffered saline or cell culture medium. The aim of this study is to develop a simple method for wetting poly (lactic-co-glycolic) acid microspheres for cell delivery applications. A one-step ethanol immersion process that involved addition of serum-supplemented medium and ethanol to PLGA microspheres over 30 min–24 h is described in the present study. This protocol presents a more efficient methodology than conventional two-step wetting procedures. Attachment of human skeletal myoblasts to poly (lactic-co-glycolic) acid microspheres was dependent on extent of wetting, changes in surface topography mediated by ethanol pre-wetting and serum protein adsorption. Ethanol, at 70% (v/v) and 100%, facilitated similar levels of wetting. Wetting with 35% (v/v) ethanol was only achieved after 24 h. Pre-wetting (over 3 h) with 70% (v/v) ethanol allowed significantly greater ( p ≤ 0.01) serum protein adsorption to microspheres than wetting with 35% (v/v) ethanol. On serum protein-loaded microspheres, greater numbers of myoblasts attached to constructs wetted with 70% ethanol than those partially wetted with 35% (v/v) ethanol. Microspheres treated with 70% (v/v) ethanol presented a more rugose surface than those treated with 35% (v/v) ethanol, indicating that more efficient myoblast adhesion to the former may be at least partially attributed to differences in surface structure. We conclude that our novel protocol for pre-wetting poly (lactic-co-glycolic) acid microspheres that incorporates biochemical and structural features into this biomaterial can facilitate myoblast delivery for use in clinical settings.

Flavonoids reduce cardiovascular disease risk through anti-inflammatory, anti-coagulant and anti-platelet actions. One key flavonoid inhibitory mechanism is blocking kinase activity that drives these processes. Flavonoids attenuate activities of kinases including phosphoinositide-3-kinase, Fyn, Lyn, Src, Syk, PKC, PIM1/2, ERK, JNK and PKA. X-ray crystallographic analyses of kinase-flavonoid complexes show that flavonoid ring systems and their hydroxyl substitutions are important structural features for their binding to kinases. A clearer understanding of structural interactions of flavonoids with kinases is necessary to allow construction of more potent and selective counterparts. We examined flavonoid (quercetin, apigenin and catechin) interactions with Src family kinases (Lyn, Fyn and Hck) applying the Sybyl docking algorithm and GRID. A homology model (Lyn) was used in our analyses to demonstrate that high-quality predicted kinase structures are suitable for flavonoid computational studies. Our docking results revealed potential hydrogen bond contacts between flavonoid hydroxyls and kinase catalytic site residues. Identification of plausible contacts indicated that quercetin formed the most energetically stable interactions, apigenin lacked hydroxyl groups necessary for important contacts and the non-planar structure of catechin could not support predicted hydrogen bonding patterns. GRID analysis using a hydroxyl functional group supported docking results. Based on these findings, we predicted that quercetin would inhibit activities of Src family kinases with greater potency than apigenin and catechin. We validated this prediction using in vitro kinase assays. We conclude that our study can be used as a basis to construct virtual flavonoid interaction libraries to guide drug discovery using these compounds as molecular templates.

Cell therapy involves the delivery of cells into damaged or diseased tissue to induce regeneration or repair. The direct transplantation of cells without carrier systems, however, results in the vast majority dying rapidly once they are placed in the body. Limitations encountered with direct cell infusions may be eliminated by their local delivery using biomaterial carrier systems. Cell encapsulation in appropriate biomaterials is a promising approach for long-term delivery of therapeutic agents. The biomaterial-based methodology for cell therapy has proven successful for the regeneration of a number of tissues including bone, myocardium, cornea, and liver. Alginate hydrogels in particular, have received considerable attention for the therapeutic delivery of cells and for the formation of tissue intended for transplantation. Advantages of alginate gels include easily tuneable biophysical properties, immunoisolatory abilities, and excellent biocompatibility. The biomechanical and biochemical properties of alginate gels can be modified to create biomimetic systems which direct stem cell phenotype, support in situ tissue engineering and preserve therapeutic cells. In this chapter we describe the versatility of alginate encapsulation technology in the context of its application to cell-based therapies. We focus on cell types that are at the preclinical stages of therapeutic application.

Noise stress (NS) is detrimental to many aspects of human health and behavior. Understanding the effect of noise stressors on human cognitive function is a growing area of research and is crucial to helping clinical populations, such as those with schizophrenia, which are particularly sensitive to stressors. A review of electronic databases for studies assessing the effect of acute NS on cognitive functions in healthy adults revealed 31 relevant studies. The review revealed (1) NS exerts a clear negative effect on attention, working memory and episodic recall, and (2) personality characteristics, in particular neuroticism, and sleep influence the impact of noise stressors on performance in interaction with task complexity. Previous findings of consistent impairment in NS-relevant cognitive domains, heightened sensitivity to stressors, elevated neuroticism and sleep disturbances in schizophrenia, taken together with the findings of this review, highlight the need for empirical studies to elucidate whether NS, a common aspect of urban environments, exacerbates cognitive deficits and other symptoms in schizophrenia and related clinical populations.

Chemical and biochemical modification of hydrogels is one strategy to create physiological constructs that maintain cell function. The aim of this study was to apply oxidised alginate hydrogels as a basis for development of a biomimetic niche for limbal epithelial stem cells that may be applied to treating corneal dysfunction. The stem phenotype of bovine limbal epithelial cells (LEC) and the viability of corneal epithelial cells (CEC) were examined in oxidised alginate gels containing collagen IV over a 3-day culture period. Oxidation increased cell viability (P ≤ 0.05) and this improved further with addition of collagen IV (P ≤ 0.01). Oxidised gels presented larger internal pores (diameter: 0.2–0.8 µm) than unmodified gels (pore diameter: 0.05–0.1 µm) and were significantly less stiff (P ≤ 0.001), indicating that an increase in pore size and a decrease in stiffness contributed to improved cell viability. The diffusion of collagen IV from oxidised alginate gels was similar to that of unmodified gels suggesting that oxidation may not affect the retention of extracellular matrix proteins in alginate gels. These data demonstrate that oxidised alginate gels containing corneal extracellular matrix proteins can influence corneal epithelial cell function in a manner that may impact beneficially on corneal wound healing therapy. © 2013 The Authors. Journal of Biomedical Materials Research Part A Published byWiley Periodicals, Inc. Part A: 102A: 3393–3400, 2014.

Limbal epithelial stem cells may ameliorate limbal stem cell deficiency through secretion of therapeutic proteins, delivered to the cornea in a controlled manner using hydrogels. In the present study the secretome of alginate-encapsulated limbal epithelial stem cells is investigated. Conditioned medium was generated from limbal epithelial stem cells encapsulated in 1.2% (w/v) calcium alginate gels. Conditioned medium proteins separated by 1-D gel electrophoresis were visualized by silver staining. Proteins of interest including secreted protein acidic and rich in cysteine, profilin-1, and galectin-1 were identified by immunoblotting. The effect of conditioned medium (from alginate-encapsulated limbal epithelial stem cells) on corneal epithelial cell proliferation was quantified and shown to significantly inhibit (P≤0.05) their growth. As secreted protein acidic and rich in cysteine was previously reported to attenuate proliferation of epithelial cells, this protein may be responsible, at least in part, for inhibition of corneal epithelial cell proliferation. We conclude that limbal epithelial stem cells encapsulated in alginate gels may regulate corneal epithelialisation through secretion of inhibitory proteins.