Mechanochemical Reactivity of a 1,2,4-Triazoline-3,5-dione-Anthracene Diels-Alder Adduct.

Force-responsive molecules that produce fluorescent moieties under stress provide a means for stress-sensing and material damage assessment. In this work, we report a mechanophore based on Diels-Alder adduct TAD-An of 4,4'-(4,4'-diphenylmethylene)-bis-(1,2,4-triazoline-3,5-dione) and initiator-substituted anthracene that can undergo retro-Diels-Alder (rDA) reaction by pulsed ultrasonication and compressive activation in bulk materials. The influence of having C-N versus C-C bonds at the sites of bond scission is elucidated by comparing the relative mechanical strength of TAD-An to another Diels-Alder adduct MAL-An obtained from maleimide and anthracene. The susceptibility to undergo rDa reaction correlates well with bond energy, such that C-N bond containing TAD-An degrades faster C-C bond containing MAL-An because C-N bond is weaker than C-C bond. Specifically, the results from polymer degradation kinetics under pulsed ultrasonication shows that polymer containing TAD-An has a rate constant of 1.59×10-5  min-1 , while MAL-An (C-C bond) has a rate constant of 1.40×10-5  min-1 . Incorporation of TAD-An in a crosslinked polymer network demonstrates the feasibility to utilize TAD-An as an alternative force-responsive probe to visualize mechanical damage where fluorescence can be "turned-on" due to force-accelerated retro-Diels-Alder reaction.

Inkjet-printed morphogenesis of tumor-stroma interface using bi-cellular bioinks of collagen-poly(N-isopropyl acrylamide-co-methyl methacrylate) mixture.

Recent advances in biomaterials and 3D printing/culture methods enable various tissue-engineered tumor models. However, it is still challenging to achieve native tumor-like characteristics due to lower cell density than native tissues and prolonged culture duration for maturation. Here, we report a new method to create tumoroids with a mechanically active tumor-stroma interface at extremely high cell density. This method, named "inkjet-printed morphogenesis" (iPM) of the tumor-stroma interface, is based on a hypothesis that cellular contractile force can significantly remodel the cell-laden polymer matrix to form densely-packed tissue-like constructs. Thus, differential cell-derived compaction of tumor cells and cancer-associated fibroblasts (CAFs) can be used to build a mechanically active tumor-stroma interface. In this methods, two kinds of bioinks are prepared, in which tumor cells and CAFs are suspended respectively in the mixture of collagen and poly (N-isopropyl acrylamide-co-methyl methacrylate) solution. These two cellular inks are inkjet-printed in multi-line or multi-layer patterns. As a result of cell-derived compaction, the resulting structure forms tumoroids with mechanically active tumor-stroma interface at extremely high cell density. We further test our working hypothesis that the morphogenesis can be controlled by manipulating the force balance between cellular contractile force and matrix stiffness. Furthermore, this new concept of "morphogenetic printing" is demonstrated to create more complex structures beyond current 3D bioprinting techniques.

Synthesis of patterned polyHIPE-hydrogel composite materials using thiol-ene chemistry.

Elastomeric materials combining multiple properties within a single composite are highly desired in applications including biomaterials interfaces, actuators, and soft robotics. High spatial resolution is required to impart different properties across the composite for the intended application, but many techniques used to prepare these composites rely on multistep and complex methods. There is a need for the development of simple and efficient platforms to design layered composite materials. Here, we report the synthesis of horizontally- and vertically-patterned composites consisting of PDMS-based polymerized high internal phase emulsion (polyHIPE) porous elastomers and PDMS/PEG hydrogels. Composites with defined interfaces that were mechanically robust were prepared, and rheological analysis of the polyHIPE and hydrogel layers showed storage moduli values of âˆ¼ 35 kPa and 45 kPa respectively. The compressive Young's Modulus and maximum strain of the polyHIPEs were dependent on the thiol to ene ratio in the formulation and obtained values ranging from 6 to 25 kPa and 50-65% respectively. The mechanical properties, total porosity of the polyHIPE, and swelling ratio of the hydrogel were unaffected by the patterning technique compared to non-patterned controls. PolyHIPE-hydrogel composite materials having up to 7-different horizontally pattered layers could be prepared that could expand and contract up hydration and drying.

Designing Interfacial Reactions for Nanometer-Scale Surface Patterning of PDMS with Controlled Elastic Modulus.

Control over the surface chemistry of elastomers such as polydimethylsiloxane (PDMS) is important for many applications. However, achieving nanostructured chemical control on amorphous material interfaces below the length scale of substrate heterogeneity is not straightforward, and can be particularly difficult to decouple from changes in network structure that are required for certain applications (e.g., variation of elastic modulus for cell culture). We have recently reported a new method for precisely structured surface functionalization of PDMS and other soft materials, which displays high densities of ligands directly on the material surface, maximizing steric accessibility. Here, we systematically examine structural factors in the PDMS components (e.g., base and cross-linker structures) that impact efficiency of the interfacial reaction that leads to surface functionalization. Applying this understanding, we demonstrate routes for generating equivalent nanometer-scale functional patterns on PDMS with elastic moduli from 0.013 to 1.4 MPa, establishing a foundation for use in applications such as cell culture.

Special considerations for studies of extracellular vesicles from parasitic helminths: A community-led roadmap to increase rigour and reproducibility.

Over the last decade, research interest in defining how extracellular vesicles (EVs) shape cross-species communication has grown rapidly. Parasitic helminths, worm species found in the phyla Nematoda and Platyhelminthes, are well-recognised manipulators of host immune function and physiology. Emerging evidence supports a role for helminth-derived EVs in these processes and highlights EVs as an important participant in cross-phylum communication. While the mammalian EV field is guided by a community-agreed framework for studying EVs derived from model organisms or cell systems [e.g., Minimal Information for Studies of Extracellular Vesicles (MISEV)], the helminth community requires a supplementary set of principles due to the additional challenges that accompany working with such divergent organisms. These challenges include, but are not limited to, generating sufficient quantities of EVs for descriptive or functional studies, defining pan-helminth EV markers, genetically modifying these organisms, and identifying rigorous methodologies for in vitro and in vivo studies. Here, we outline best practices for those investigating the biology of helminth-derived EVs to complement the MISEV guidelines. We summarise community-agreed standards for studying EVs derived from this broad set of non-model organisms, raise awareness of issues associated with helminth EVs and provide future perspectives for how progress in the field will be achieved.

Self-Assembled Asperities for Pressure-Tunable Adhesion.

Control of adhesion is important in a host of applications including soft robotics, pick-and-place manufacturing, wearable devices, and transfer printing. While there are adhesive systems with discrete switchability between states of high and low adhesion, achieving continuously variable adhesion strength remains a challenge. In this work, a pressure-tunable adhesive (PTA) that is based on the self-assembly of stiff microscale asperities on an elastomeric substrate is presented. It is demonstrated that the adhesion strength of the PTA increases with the applied compressive preload due to the unique contact formation mechanism caused by the asperities. Additionally, a contact mechanics model is developed to explain the resulting trends. For a specific PTA design, the critical pull-off force can be increased from 0.4 to 30 mN by increasing the applied preload from 1 to 30 mN. Finally, the applicability of precision control of adhesion strength is demonstrated by utilizing the PTA for pick-and-place material handling. The approach in pressure-tunable adhesive design based on self-assembly of asperities presents a scalable and versatile approach that is applicable to a variety of material systems having different mechanical or surface properties.

Systemic Corticosteroids in the Management of Pediatric Cystic Fibrosis Pulmonary Exacerbations.

Rationale: Pulmonary exacerbation (PEx) events contribute to lung function decline in people with cystic fibrosis (CF). CF Foundation PEx guidelines note that a short course of systemic corticosteroids may offer benefit without contributing to long-term adverse effects. However, insufficient evidence exists to recommend systemic corticosteroids for PEx treatment. Objectives: To determine if systemic corticosteroids for the treatment of in-hospital pediatric PEx are associated with improved clinical outcomes compared with treatment without systemic corticosteroids. Methods: We conducted a retrospective cohort study using the CF Foundation Patient Registry-Pediatric Health Information System linked database. People with CF were included if hospitalized for a PEx between 2006 and 2018 and were 6-21 years of age. Time to next PEx was assessed by Cox proportional hazards regression. Lung function outcomes were assessed by linear mixed-effect modeling and generalized estimating equations. To address confounding by indication, inverse probability treatment weighting was used. Results: A total of 3,471 people with CF contributed 9,787 PEx for analysis. Systemic corticosteroids were used in 15% of all PEx. In our primary analysis, systemic corticosteroids were not associated with better pre- to post-PEx percent predicted forced expiratory volume in 1 second responses (mean difference, -0.36; 95% confidence interval [CI], -1.14, 0.42; P = 0.4) or a higher odds of returning to lung function baseline (odds ratio, 0.97; 95% CI, 0.84-1.12; P = 0.7) but were associated with a reduced chance of future PEx requiring intravenous antibiotics (hazard ratio, 0.91; 95% CI, 0.85-0.96; P = 0.002). When restricting the analysis to one PEx per person, lung function outcomes remained no different among PEx treated with or without systemic corticosteroids, but, in contrast to our primary analysis, the use of systemic corticosteroids was no longer associated with a reduced chance of having a future PEx requiring intravenous antibiotics (hazard ratio, 0.96; 95% CI, 0.86, 1.07; P = 0.42). Conclusions: Systemic corticosteroid treatment for in-hospital pediatric PEx was not associated with improved lung function outcomes. Prospective trials are needed to better evaluate the risks and benefits of systemic corticosteroid use for PEx treatment in children with CF.

Associations between Gastrointestinal Nematode Infection Burden and Lying Behaviour as Measured by Accelerometers in Periparturient Ewes.

The application of precision livestock farming (PLF) technologies will underpin new strategies to support the control of livestock disease. However, PLF technology is underexploited within the sheep industry compared to other livestock sectors, and research is essential to identify opportunities for PLF applications. These opportunities include the control of endemic sheep disease such as parasitic gastroenteritis, caused by gastrointestinal nematode infections, which is estimated to cost the European sheep industry EUR 120 million annually. In this study, tri-axial accelerometers recorded the behaviour of 54 periparturient Welsh Mule ewes to discover if gastrointestinal nematode (GIN) infection burden, as measured by faecal egg count (FEC), was associated with behavioural variation. Linear mixed models identified that increasing FECs in periparturient ewes were significantly associated with a greater number of lying bouts per day and lower bout durations (p = 0.013 and p = 0.010, respectively). The results demonstrate that FECs of housed periparturient ewes are associated with detectable variations in ewe behaviour, and as such, with further investigation there is potential to develop future targeted selective treatment protocols against GIN in sheep based on behaviour as measured by PLF technologies.

The effect of postprandial exercise on mean blood glucose concentrations following high and maintenance carbohydrate content meals in healthy dogs.

OBJECTIVES: To compare the effect of 15 min of exercise 30 min post-meal on mean blood glucose concentrations in 5 well-conditioned versus 5 over-conditioned dogs. To compare the effect of exercise on glycemic control in dogs eating their maintenance diet as compared to a high carbohydrate meal. ANIMALS: Ten healthy staff or student owned dogs, five well- and five over-conditioned. PROCEDURES: This was a crossover study over 5 days. Continuous glucose monitors (CGM) were placed on day 1. On days 2 and 3, dogs received their maintenance diet and a high carbohydrate meal, respectively and were walked on the treadmill for 15 min following each meal. On day 4, dogs were given their maintenance diet in hospital without treadmill activity. On day 5, the CGM were removed. The mean blood glucose 30 min post-meal, during exercise, 15 min after completing exercise, and the 1-3 h period after completing the exercise were compared to detect any effect of exercise, diet composition, or body condition. RESULTS: Dogs consuming a high carbohydrate meal had a significantly higher mean blood glucose 15 min post-exercise. Mean glucose values at all time points following a high carbohydrate meal were significantly higher than mean glucose values on the non-exercise day. CONCLUSIONS AND CLINICAL RELEVANCE: No impact of post-prandial exercise on glucose concentrations were identified in this study, however, the carbohydrate content of the meal impacted post-prandial glycemic responses in healthy dogs regardless of body condition. Evaluating the impact of post-prandial exercise in insulin-dependent or glucose-intolerant dogs is warranted to determine if these findings persist.

Activation of Mechanophores in a Thermoset Matrix by Instrumented Scratch.

Scratches in polymer coatings and barrier layers negatively impact optical properties (haze, light transmission, etc.), initiate routes of degradation or corrosion (moisture permeability), and nucleate delamination of the coating. Detecting scratches in coatings on advanced materials systems is an important component of structural health monitoring but can be difficult if the defects are too small to be detected by the naked eye. The primary focus of the present work is to investigate scratch damage using fluorescence lifetime imaging microscopy (FLIM) and mechanical activation of a mechanophore (MP)-containing transparent epoxy coating. The approach utilizes a Berkovich tip to scratch MP-epoxy coatings under a linearly increasing normal load. The goal is to utilize the fluorescent behavior of activated MPs to enable the detection of microscale scratches and molecular scale changes in polymeric systems. Taking advantage of the amine functionality present in a polyetheramine/bisphenol A epoxy network, a modified rhodamine dye is covalently bonded into a transparent, thermoset polymer network. Following instrumented scratch application, subsequent fluorescence imaging of the scratched MP-epoxy reveals the extent of fluorescence activation induced by the mechanical deformation. In this work, the rhodamine-based mechanophore is used to identify both ductile and fracture-dominated processes during the scratch application. The fluorescence intensity increases linearly with the applied normal load and is sensitive to fracture dominated processes. Fluorescence lifetime and hyperspectral imaging of damage zones provide additional insight into the local (nanoscopic) environment and molecular structure of the MP around the fracture process zone, respectively. The mechanophore/scratch deformation approach allows a fluorescence microscope to probe local yielding and fracture events in a powerful way that enhances the optical characterization of damage zones formed by standard scratch test methods and leads to novel defect detection strategies.

Evidence of Immune Modulators in the Secretome of the Equine Tapeworm Anoplocephala perfoliata.

Anoplocephala perfoliata is a neglected gastro-intestinal tapeworm, commonly infecting horses worldwide. Molecular investigation of A. perfoliata is hampered by a lack of tools to better understand the host-parasite interface. This interface is likely influenced by parasite derived immune modulators released in the secretome as free proteins or components of extracellular vesicles (EVs). Therefore, adult RNA was sequenced and de novo assembled to generate the first A. perfoliata transcriptome. In addition, excretory secretory products (ESP) from adult A. perfoliata were collected and EVs isolated using size exclusion chromatography, prior to proteomic analysis of the EVs, the EV surface and EV depleted ESP. Transcriptome analysis revealed 454 sequences homologous to known helminth immune modulators including two novel Sigma class GSTs, five α-HSP90s, and three α-enolases with isoforms of all three observed within the proteomic analysis of the secretome. Furthermore, secretome proteomics identified common helminth proteins across each sample with known EV markers, such as annexins and tetraspanins, observed in EV fractions. Importantly, 49 of the 454 putative immune modulators were identified across the secretome proteomics contained within and on the surface of EVs in addition to those identified in free ESP. This work provides the molecular tools for A. perfoliata to reveal key players in the host-parasite interaction within the horse host.

Design and validation of a modular micro-robotic system for the mechanical characterization of soft tissues.

The mechanical properties of tissues are critical design parameters for biomaterials and regenerative therapies seeking to restore functionality after disease or injury. Characterizing the mechanical properties of native tissues and extracellular matrix throughout embryonic development helps us understand the microenvironments that promote growth and remodeling, activities critical for biomaterials to support. The mechanical characterization of small, soft materials like the embryonic tissues of the mouse, an established mammalian model for development, is challenging due to difficulties in handling minute geometries and resolving forces of low magnitude. While uniaxial tensile testing is the physiologically relevant modality to characterize tissues that are loaded in tension in vivo, there are no commercially available instruments that can simultaneously measure sufficiently low tensile force magnitudes, directly measure sample deformation, keep samples hydrated throughout testing, and effectively grip minute geometries to test small tissues. To address this gap, we developed a micromanipulator and spring system that can mechanically characterize small, soft materials under tension. We demonstrate the capability of this system to measure the force contribution of soft materials, silicone, fibronectin sheets, and fibrin gels with a 5 nN - 50 µN force resolution and perform a variety of mechanical tests. Additionally, we investigated murine embryonic tendon mechanics, demonstrating the instrument can measure differences in mechanics of small, soft tissues as a function of developmental stage. This system can be further utilized to mechanically characterize soft biomaterials and small tissues and provide physiologically relevant parameters for designing scaffolds that seek to emulate native tissue mechanics. STATEMENT OF SIGNIFICANCE: The mechanical properties of cellular microenvironments are critical parameters that contribute to the modulation of tissue growth and remodeling. The field of tissue engineering endeavors to recapitulate these microenvironments in order to construct tissues de novo. Therefore, it is crucial to uncover the mechanical properties of the cellular microenvironment during tissue formation. Here, we present a system capable of acquiring microscale forces and optically measuring sample deformation to calculate the stress-strain response of soft, embryonic tissues under tension, and easily adaptable to accommodate biomaterials of various sizes and stiffnesses. Altogether, this modular system enables researchers to probe the unknown mechanical properties of soft tissues throughout development to inform the engineering of physiologically relevant microenvironments.

Temporal dynamics of trematode intermediate snail host environmental DNA in small water body habitats.

Environmental DNA (eDNA) surveying has potential to become a powerful tool for sustainable parasite control. As trematode parasites require an intermediate snail host that is often aquatic or amphibious to fulfil their lifecycle, water-based eDNA analyses can be used to screen habitats for the presence of snail hosts and identify trematode infection risk areas. The aim of this study was to identify climatic and environmental factors associated with the detection of Galba truncatula eDNA. Fourteen potential G. truncatula habitats on two farms were surveyed over a 9-month period, with eDNA detected using a filter capture, extraction and PCR protocol with data analysed using a generalized estimation equation. The probability of detecting G. truncatula eDNA increased in habitats where snails were visually detected, as temperature increased, and as water pH decreased (P < 0.05). Rainfall was positively associated with eDNA detection in watercourse habitats on farm A, but negatively associated with eDNA detection in watercourse habitats on farm B (P < 0.001), which may be explained by differences in watercourse gradient. This study is the first to identify factors associated with trematode intermediate snail host eDNA detection. These factors should be considered in standardized protocols to evaluate the results of future eDNA surveys.

Influence of alignment and microstructure features on the mechanical properties and failure mechanisms of cellulose nanocrystals (CNC) films.

The mechanical properties of cellulose nanocrystal (CNC) films critically depend on many microstructural parameters such as fiber length distribution (FLD), fiber orientation distribution (FOD), and the strength of the interactions between the fibers. In this paper, we use our coarse-grained molecular model of CNC to study the effect of length and orientation distribution and attractions between CNCs on the mechanical properties of neat CNCs. The effect of misalignment of a 2D staggered structure of CNC with respect to the loading direction was studied with simulations and analytical solutions and then verified with experiments. To understand the effect of FLD and FOD on the mechanical performance, various 3D microstructures representing different case studies such as highly aligned, randomly distributed, short length CNCs and long length CNCs were generated and simulated. According to the misalignment study, three different failure modes: sliding mode, mixed mode, and normal mode were defined. Also, comparing the effects of FOD, FLD, and CNC interaction strength, shows that the adhesion strength is the only parameter that can significantly improve the mechanical properties, regardless of loading direction or FOD of CNCs.

Interruptions to Intensive Care Nurses and Clinical Errors and Procedural Failures: A Controlled Study of Causal Connection.

OBJECTIVES: Interruptions occur frequently in the intensive care unit (ICU) and are associated with errors. To date, no causal connection has been established between interruptions and errors in healthcare. It is important to know whether interruptions directly cause errors before implementing interventions designed to reduce interruptions in ICUs. The aim of the study was to investigate whether ICU nurses who receive a higher number of workplace interruptions commit more clinical errors and procedural failures than those who receive a lower number of interruptions. METHODS: We conducted a prospective controlled trial in a high-fidelity ICU simulator. A volunteer sample of ICU nurses from a single unit prepared and administered intravenous medications for a patient manikin. Nurses received either 3 (n = 35) or 12 (n = 35) scenario-relevant interruptions and were allocated to either condition in an alternating fashion. Primary outcomes were the number of clinical errors and procedural failures committed by each nurse. RESULTS: The rate ratio of clinical errors committed by nurses who received 12 interruptions compared with nurses who received 3 interruptions was 2.0 (95% confidence interval = 1.41-2.83, P < 0.001). The rate ratio of procedural failures committed by nurses who received 12 interruptions compared with nurses who were interrupted 3 times was 1.2 (95% confidence interval = 1.05-1.37, P = 0.006). CONCLUSIONS: More workplace interruptions during medication preparation and administration lead to more clinical errors and procedural failures. Reducing the frequency of interruptions may reduce the number of errors committed; however, this should be balanced against important information that interruptions communicate.

Pressure tunable adhesion of rough elastomers.

The ability to control adhesion is critical in various technologies including wearable electronics, pressure sensitive adhesives, and robotic systems. Biomimetic fibrillar structures, random surface roughness, and chemical surface treatments have been employed to modify the adhesion energy of materials used in these applications. However, polymer thin film dewetting has not been investigated as a surface modification tool to control adhesion. In this work, polystyrene thin films are thermally annealed on a polydimethylsiloxane substrate, causing them to dewet and form stiff, microscopic asperities on the soft substrate. The size of the asperities increases with increasing pre-annealing film thickness. Adhesion is quantified by flat-punch normal indentation testing. The largest asperities exhibited a decrease in adhesion to below the sensitivity of the instrument. More interestingly, the surfaces covered with the smallest asperities displayed a pressure-dependent adhesive response. By increasing the normal compressive stress applied prior to separation, the total debonding energy increased monotonically on the smallest asperity-covered surfaces.

Rapid detection of Galba truncatula in water sources on pasture-land using loop-mediated isothermal amplification for control of trematode infections.

BACKGROUND: Fascioliasis caused by the trematodes Fasciola hepatica and F. gigantica, is a global neglected zoonotic disease estimated to cost the livestock industry over €2.5 billion annually. Farm management measures and sustainable use of anthelmintics can, in principle, effectively control trematode infection in livestock and reduce the rate of developing anthelmintic resistance. Previously, we designed an environmental DNA (eDNA) assay to identify a common trematode intermediate host, the freshwater snail Galba truncatula, in water sources to measure specific trematode infection risk areas on pasture-land. To improve this procedure, we now report a loop-mediated isothermal amplification (LAMP) assay to identify G. truncatula eDNA. METHODS: A LAMP assay was designed and optimised (e.g. temperature, time duration and primer concentration) to identify G. truncatula DNA. The ability of the LAMP assay to target G. truncatula DNA was identified, and LAMP assay limit of detection was investigated in comparison to conventional PCR. In the field, 48 water samples were collected from stream, ditch and water pool habitats in four locations at two Aberystwyth University farms over a seven week period to investigate the applicability of the LAMP assay for use on eDNA samples, in comparison to conventional PCR. RESULTS: The LAMP assay delivered detectable results in 30 min at 63 °C. The assay discriminated between G. truncatula DNA and non-target DNA, presenting a level of DNA detection comparable to conventional PCR. No significant difference was found between the ability of the LAMP and PCR assay to identify G. truncatula eDNA in water samples. Kappa coefficient analysis revealed a moderate level of agreement between LAMP and PCR assays. CONCLUSIONS: This study demonstrated that the LAMP assay can detect G. truncatula eDNA in a simple and rapid manner. The LAMP assay may become a valuable tool to determine optimum pasture management for trematode parasite control.

Evidence of sequestration of triclabendazole and associated metabolites by extracellular vesicles of Fasciola hepatica.

Fascioliasis is a neglected zoonotic disease that infects humans and ruminant species worldwide. In the absence of vaccines, control of fascioliasis is primarily via anthelminthic treatment with triclabendazole (TCBZ). Parasitic flatworms, including Fasciola hepatica, are active secretors of extracellular vesicles (EVs), but research has not been undertaken investigating EV anthelmintic sequestration. Adult F. hepatica were cultured in lethal and sub-lethal doses of TCBZ and its active metabolites, in order to collect EVs and evaluate their morphological characteristics, production and anthelmintic metabolite content. Transmission electron microscopy demonstrated that F. hepatica exposed to TCBZ and its metabolites produced EVs of similar morphology, compared to non-TCBZ exposed controls, even though TCBZ dose and/or TCBZ metabolite led to measurable structural changes in the treated F. hepatica tegument. qNano particle analysis revealed that F. hepatica exposed to TCBZ and its metabolites produced at least five times greater EV concentrations than non-TCBZ controls. A combined mass spectrometry and qNano particle analysis confirmed the presence of TCBZ and the TCBZ-sulphoxide metabolite in anthelmintic exposed EVs, but limited TCBZ sulphone was detectable. This data suggests that EVs released from adult F. hepatica have a biological role in the sequestration of TCBZ and additional toxic xenobiotic metabolites.