Bioaccumulation of pollutants in the green-lipped mussel Perna viridis: Assessing pollution abatement in Victoria Harbour and its adjacent aquaculture area, Hong Kong, and the minimal human health risks from mussel consumption.

The green-lipped mussel Perna viridis was utilised for pollution biomonitoring in Victoria Harbour and its adjacent aquaculture area in Hong Kong. P. viridis was collected from a reference site and redeployed at five study sites for five weeks during the dry and wet seasons of 2019. Our study found various polycyclic aromatic hydrocarbons (PAHs) and heavy metals in the mussel tissue, while polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) were not detected. P. viridis at the reference site generally displayed lower levels of pollutants. Comparing with previous research in the 1980s and 2000s, we observed substantial reduction in the tissue levels of PAHs, PCBs, OCPs and heavy metals in P. viridis. The human health risks associated with consuming these mussels were determined to be insignificant. Our findings imply that the Harbour Area Treatment Scheme has been effective in improving the water quality in Victoria Harbour and its adjacent aquaculture area.

Surgeon-designed patient-specific instrumentation improves glenoid component screw placement for reverse total shoulder arthroplasty in a population with small glenoid dimensions.

PURPOSE: Glenoid component loosening is a potential complication of reverse total shoulder arthroplasty (rTSA), occurring in part due to lack of adequate screw purchase in quality scapular bone stock. This study was to determine the efficacy of a surgeon-designed, 3D-printed patient-specific instrumentation (PSI) compared to conventional instrumentation (CI) in achieving longer superior and inferior screw lengths for glenoid component fixation. METHODS: A multi-centre retrospective analysis of patients who underwent rTSA between 2015 and 2020. Lengths of the superior and inferior locking screws inserted for fixation of the glenoid baseplate component were recorded and compared according to whether patients received PSI or CI. Secondary outcomes included operative duration and incidence of complications requiring revision surgery. RESULTS: Seventy-three patients (31 PSI vs. 42 CI) were analysed. Average glenoid diameter was 24.5 mm (SD: 3.1) and 81% of patients had smaller glenoid dimensions compared to the baseplate itself. PSI produced significantly longer superior (44.7 vs. 30.7 mm; P < 0.001) and inferior (43.0 vs. 31 mm; P < 0.001) mean screw lengths, as compared to CI. A greater proportion of maximal screw lengths for the given rTSA construct (48 mm) were observed in the PSI group (71.9% vs. 11.9% superior, 59.4% vs. 11.9% inferior). Operative duration was not statistically significantly different between the PSI and CI groups (150 min vs. 169 min, respectively; P = 0.229). No patients had radiographic loosening of the glenoid component with an average of 2-year follow-up. CONCLUSION: PSI facilitates longer superior and inferior screw placement in the fixation of the glenoid component for rTSA. With sufficient training, PSI can be designed and implemented by surgeons themselves.

Adverse impacts of high-density microplastics on juvenile growth and behaviour of the endangered tri-spine horseshoe crab Tachypleus tridentatus.

The impacts of high-density microplastics, namely polyamine 6,6 (nylon), polymethyl methacrylate (PMMA) and polyethylene terephthalate (PET), on growth and behaviour of the endangered tri-spine horseshoe crab Tachypleus tridentatus were investigated for 100 days. Negative changes in wet weight and prosomal width of the juveniles were observed in all treatments of microplastics, but significant difference was only detected in prosomal width between control and PMMA. T. tridentatus became significantly less active upon exposure to nylon and PET. The extent of burrowing by T. tridentatus did not significantly differ among the treatments but was overall significantly reduced towards day 100. T. tridentatus exposed to PET significantly showed the lowest survival probability (30 %), compared to the other treatments (70-90 %). In conclusion, high-density microplastics compromised growth and behaviour of juvenile horseshoe crabs. Among the polymers that were tested, PET was considered more harmful and associated with higher mortality.

A short review on the recent method development for extraction and identification of microplastics in mussels and fish, two major groups of seafood.

The prevalence of microplastics in the marine environment poses potential health risks to humans through seafood consumption. Relevant data are available but the diverse analytical approaches adopted to characterise microplastics have hampered data comparison among studies. Here, the techniques for extraction and identification of microplastics are summarised among studies of marine mussels and fish, two major groups of seafood. Among the reviewed papers published in 2018-2021, the most common practice to extract microplastics was through tissue digestion in alkaline chemicals (46 % for mussels, 56 % for fish) and oxidative chemicals (28 % for mussels, 12 % for fish). High-density solutions such as sodium chloride could be used to isolate microplastics from other undigested residues by flotation. Polymer analysis of microplastics was mainly carried out with Fourier-transform infrared (FTIR) spectroscopy (58 % for both mussels and fish) and Raman spectroscopy (14 % for mussels, 8 % for fish). Among these methods, we recommend alkaline digestion for microplastic extraction, and the automated mapping approach of FTIR or Raman spectroscopy for microplastic identification. Overall, this study highlights the need for a standard protocol for characterising microplastics in seafood samples.

3D printed auxetic heel pads for patients with diabetic mellitus.

More than 422 million people worldwide suffered from diabetes mellitus (DM) in 2021. Diabetic foot is one the most critical complications resultant of DM. Foot ulceration and infection are frequently arisen, which are associated with changes in the mechanical properties of the plantar soft tissues, peripheral arterial disease, and sensory neuropathy. Diabetic insoles are currently the mainstay in reducing the risk of foot ulcers by reducing the magnitude of the pressure on the plantar Here, we propose a novel pressure relieving heel pad based on a circular auxetic re-entrant honeycomb structure by using three-dimensional (3D) printing technology to minimize the pressure on the heel, thus reducing the occurrence of foot ulcers. Finite element models (FEMs) are developed to evaluate the structural changes of the developed circular auxetic structure upon exertion of compressive forces. Moreover, the effects of the internal angle of the re-entrant structure on the peak contact force and the mean pressure acting on the heel as well as the contact area between the heel and the pads are investigated through a finite element analysis (FEA). Based on the result from the validated FEMs, the proposed heel pad with an auxetic structure demonstrates a distinct reduction in the peak contact force (∼10%) and the mean pressure (∼14%) in comparison to a conventional diabetic insole (PU foam). The characterized result of the designed circular auxetic structure not only provides new insights into diabetic foot protection, but also the design and development of various impact resistance products.

Patient-specific 3D-printed helmet for post-craniectomy defect - a case report.

BACKGROUND: Patients who undergo decompressive craniectomy (DC) are often fitted with a helmet that protects the craniectomy site from injury during rehabilitation. However, conventional "one-size-fits-all" helmets may not be feasible for certain craniectomy defects. We describe the production and use of a custom 3D-printed helmet for a DC patient where a conventional helmet was not feasible due to the craniectomy defect configuration. CASE PRESENTATION: A 65-year-old male with ethmoid sinonasal carcinoma underwent cranionasal resection and DC with free vastus lateralis flap reconstruction to treat cerebrospinal fluid leakage. He required an external helmet to protect the craniectomy site, however, the rim of a conventional helmet compressed the craniectomy site, and the straps compressed the vascular pedicle of the muscle flap. Computed topography (CT) scans of the patient's cranium were imported into 3D modelling software and used to fabricate a patient-specific, strapless helmet using fused deposition modelling (FDM). The final helmet fit the patient perfectly and circumvented the compression issues, while also providing better cosmesis than the conventional helmet. Four months postoperatively, the helmet remains intact and in use. CONCLUSIONS: 3D printing can be used to produce low-volume, patient-specific external devices for rehabilitation where standardized adjuncts are not optimal. Once initial start-up costs and training are overcome, these devices can be produced by surgeons themselves to meet a wide range of clinical needs.

Determination of microplastics in the edible green-lipped mussel Perna viridis using an automated mapping technique of Raman microspectroscopy.

Microplastics are prevalent in marine environments and seafood and thus can easily end up in human diets. This has raised serious concerns worldwide, particularly in Hong Kong where the seafood consumption per capita can be three times higher than the global average. This study focused on the green-lipped mussel Perna viridis, a popular seafood species which is subject to a high risk of contamination by microplastics due to its filter-feeding nature. P. viridis was collected from five mariculture sites in Hong Kong and assessed for its body load of microplastics using an automated Raman mapping approach. Microplastics were found in all sites, with an average of 1.60-14.7 particles per mussel per site, or 0.21-1.83 particles per g wet weight. Polypropylene, polyethylene, polystyrene and polyethylene terephthalate were detected among the microplastics, mainly as fragments or fibres in the size range of 40-1000 µm. It was estimated that through consumption of P. viridis, the population in Hong Kong could ingest up to 10,380 pieces of microplastics per person per year. These estimated rates were high compared to the values reported worldwide, suggesting the potential human health risk of microplastics in Hong Kong and adjacent areas.

Improved Raman spectroscopy-based approach to assess microplastics in seafood.

Microplastics represent an emerging environmental issue and have been found almost everywhere including seafood, raising a great concern about the ecological and human health risks they pose. This study addressed the common technical challenges in the assessment of microplastics in seafood by developing an improved protocol based on Raman spectroscopy and using the green-lipped mussel Perna viridis and the Japanese jack mackerel Trachurus japonicus as the test models. Our findings identified a type of stainless-steel filter membranes with minimal Raman interference, and a combination of chemicals that achieved 99-100% digestion efficiency for both organic and inorganic biomass. This combined chemical treatment reached 90-100% recovery rates for seven types of microplastics, on which the surface modification was considered negligible and did not affect the accuracy of polymer identification based on Raman spectra, which showed 94-99% similarity to corresponding untreated microplastics. The developed extraction method for microplastics was further combined with an automated Raman mapping approach, from which our results confirmed the presence of microplastics in P. viridis and T. japonicus collected from Hong Kong waters. Identified microplastics included polypropylene, polyethylene, polystyrene and poly(ethylene terephthalate), mainly in the form of fragments and fibres. Our protocol is applicable to other biological samples, and provides an improved alternative to streamline the workflow of microplastic analysis for routine monitoring purposes.

Three-Dimensional Printing and Computer Navigation for Correction of Multiple Deformities in Osteogenesis Imperfecta: A Case Report.

CASE: A 44-year-old man with osteogenesis imperfecta presented with multiple debilitating musculoskeletal deformities. Bi-level osteotomies, assisted by 3-dimensional (3D)-printed patient-specific cutting guides, were performed to correct extraarticular valgus and procurvatum tibial deformities. Concomitant computer-navigated total knee arthroplasty was performed to restore neutral mechanical alignment. Postoperative x-ray showed good correction of deformities, and 1 year postoperatively, the patient is able to walk unaided with significant resolution of knee pain. CONCLUSION: 3D-printed osteotomy guides and computer navigation can be instrumental in procedures requiring a high degree of precision. With sufficient training, modern orthopaedic technologies can be implemented by surgeons themselves and combined to facilitate precise and personalized management of challenging conditions.

Early prognostication of COVID-19 to guide hospitalisation versus outpatient monitoring using a point-of-test risk prediction score.

INTRODUCTION: Risk factors of adverse outcomes in COVID-19 are defined but stratification of mortality using non-laboratory measured scores, particularly at the time of prehospital SARS-CoV-2 testing, is lacking. METHODS: Multivariate regression with bootstrapping was used to identify independent mortality predictors in patients admitted to an acute hospital with a confirmed diagnosis of COVID-19. Predictions were externally validated in a large random sample of the ISARIC cohort (N=14 231) and a smaller cohort from Aintree (N=290). RESULTS: 983 patients (median age 70, IQR 53-83; in-hospital mortality 29.9%) were recruited over an 11-week study period. Through sequential modelling, a five-predictor score termed SOARS (SpO2, Obesity, Age, Respiratory rate, Stroke history) was developed to correlate COVID-19 severity across low, moderate and high strata of mortality risk. The score discriminated well for in-hospital death, with area under the receiver operating characteristic values of 0.82, 0.80 and 0.74 in the derivation, Aintree and ISARIC validation cohorts, respectively. Its predictive accuracy (calibration) in both external cohorts was consistently higher in patients with milder disease (SOARS 0-1), the same individuals who could be identified for safe outpatient monitoring. Prediction of a non-fatal outcome in this group was accompanied by high score sensitivity (99.2%) and negative predictive value (95.9%). CONCLUSION: The SOARS score uses constitutive and readily assessed individual characteristics to predict the risk of COVID-19 death. Deployment of the score could potentially inform clinical triage in preadmission settings where expedient and reliable decision-making is key. The resurgence of SARS-CoV-2 transmission provides an opportunity to further validate and update its performance.

A Novel Bespoke Hypertrophic Scar Treatment: Actualizing Hybrid Pressure and Silicone Therapies with 3D Printing and Scanning.

The treatment of hypertrophic scars (HSs) is considered to be the most challenging task in wound rehabilitation. Conventional silicone sheet therapy has a positive effect on the healing process of HSs. However, the dimensions of the silicone sheet are typically larger than those of the HS itself which may negatively impact the healthy skin that surrounds the HS. Furthermore, the debonding and displacement of the silicone sheet from the skin are critical problems that affect treatment compliance. Herein, we propose a bespoke HS treatment design that integrates pressure sleeve with a silicone sheet and use of silicone gel using a workflow of three-dimensional (3D) printing, 3D scanning and computer-aided design, and manufacturing software. A finite element analysis (FEA) is used to optimize the control of the pressure distribution and investigate the effects of the silicone elastomer. The result shows that the silicone elastomer increases the amount of exerted pressure on the HS and minimizes unnecessary pressure to other parts of the wrist. Based on this treatment design, a silicone elastomer that perfectly conforms to an HS is printed and attached onto a customized pressure sleeve. Most importantly, unlimited scar treating gel can be applied as the means to optimize treatment of HSs while the silicone sheet is firmly affixed and secured by the pressure sleeve.

Microplastics from effluents of sewage treatment works and stormwater discharging into the Victoria Harbor, Hong Kong.

Sewage treatment works and stormwater outfalls were identified as sources of microplastics in the Victoria Harbor, Hong Kong. Local treated sewage and stormwater effluents contained up to 10,816 pieces per m3 of microplastics, mainly polyethylene (PE) and polypropylene (PP), being discharged at an average rate of 3.5 mg per capita daily. Bioaccumulation of microplastics in marine fish collected from the vicinity of the effluent discharge outfalls was also studied. The temporal variations of microplastics in terms of abundance, shape and polymer type in fish were found consistent with those in the effluents and surface water. The abundance of microplastics was significantly higher in March 2017 (dry season). Microplastics in fish were predominantly in fiber form and identified as PE and PP. The observed temporal variations suggest uptake of microplastics by fish from the treated sewage and stormwater effluents.

Negative expectations interfere with the analgesic effect of safety cues on pain perception by priming the cortical representation of pain in the midcingulate cortex.

It is well known that the efficacy of treatment effects, including those of placebos, is heavily dependent on positive expectations regarding treatment outcomes. For example, positive expectations about pain treatments are essential for pain reduction. Such positive expectations not only depend on the properties of the treatment itself, but also on the context in which the treatment is presented. However, it is not clear how the preceding threat of pain will bias positive expectancy effects. One hypothesis is that threatening contexts trigger fearful and catastrophic thinking, reducing the pain-relieving effects of positive expectancy. In this study, we investigated the disruptive influence of threatening contexts on positive expectancy effects while 41 healthy volunteers experienced laser-induced heat pain. A threatening context was induced using pain-threatening cues that preceded the induction of positive expectancies via subsequent pain-safety cues. We also utilised electroencephalography (EEG) to investigate potential neural mechanisms underlying these effects. Lastly, we used the Fear of Pain Questionnaire to address whether the disruptive effect of negative contexts on cued pain relief was related to the degree of fear of pain. As predicted, participants responded less to pain-safety cues (i.e., experienced more pain) when these were preceded by pain-threatening cues. In this threatening context, an enhancement of the N2 component of the laser-evoked potential was detected, which was more pronounced in fearful individuals. This effect was localised to the midcingulate cortex, an area thought to integrate negative affect with pain experience to enable adaptive behaviour in aversive situations. These results suggest that threatening contexts disrupt the effect of pain relief cues via an aversive priming mechanism that enhances neural responses in the early stages of sensory processing.

Osseointegrated implants into a variety of composite free flaps: A comparative analysis.

BACKGROUND: Significant oral function is often lost after surgical therapy for head and neck cancer. The use of osseointegrated implants for reconstruction in patients with head and neck surgery has shown to significantly improve the quality of life for these patients. Variable success rates range from 99% to 70%. METHODS: A retrospective audit of patient records was performed looking at cumulative survival of implants. Inclusion criteria were patients who were treated at 1 of 2 designated Australian Head and Neck Units and received oral osseointegrated implants. RESULTS: Fifty-nine patients were included for analysis. One hundred ninety-nine implants were placed into vascularized bone grafts (VBGs). There were 11 implant failures with an overall success rate of 94.5%. There was 1 significant adverse outcome with a pathological fracture of a flap after implant placement. Implant success in scapula and iliac crest flaps was comparable to fibula flaps. CONCLUSION: Implants placed into VBGs have a reasonable success rate in well-selected patients. © 2016 Wiley Periodicals, Inc. Head Neck 39: 443-447, 2017.

The perception of affective touch in Parkinson's disease and its relation to small fibre neuropathy.

Affective touch sensation is conducted by a sub-class of C-fibres in hairy skin known as C-Tactile (CT) afferents. CT afferents respond maximally to gentle skin stroking at velocities between 1 and 10 cm/s. Parkinson's disease (PD) is characterised by markedly reduced cutaneous C-fibres. It is not known if affective touch perception is influenced by C-fibre density and if affective touch is impaired in PD compared to healthy controls. We predicted that perceived pleasantness to gentle stroking in PD would correlate with C-afferent density and that affective touch perception would be impaired in PD compared to healthy controls. Twenty-four PD patients and 27 control subjects rated the pleasantness of brush stroking at an optimum CT stimulation velocity (3 cm/s) and two sub-optimal velocities (0.3 and 30 cm/s). PD patients underwent quantification of C-fibre density using skin biopsies and corneal confocal microscopy. All participants rated a stroking velocity of 3 cm/s as the most pleasant with significantly lower ratings for 0.3 and 30 cm/s. There was a significant positive correlation between C-fibre density and pleasantness ratings at 3 and 30 cm/s but not 0.3 cm/s. Mean pleasantness ratings were consistently higher in PD patients compared to control subjects across all three velocities. This study shows that perceived pleasantness to gentle touch correlates significantly with C-fibre density in PD. The higher perceived pleasantness in PD patients compared to controls suggests central sensitisation to peripheral inputs, which may have been enhanced by dopamine therapy.

Coupling of TRPV6 and TMEM16A in epithelial principal cells of the rat epididymis.

The epididymis establishes a congenial environment for sperm maturation and protection. Its fluid is acidic, and the calcium concentration is low and declines along the length of the epididymal tubule. However, our knowledge of ionic currents and mechanisms of calcium homeostasis in rat epididymal epithelial cells remains enigmatic. In this study, to better understand calcium regulation in the epididymis, we use the patch-clamp method to record from single rat cauda epididymal principal cells. We detect a constitutively active Ca(2+) current with characteristics that match the epithelial calcium channel TRPV6. Electrophysiological and pharmacological data also reveal a constitutively active calcium-activated chloride conductance (CaCC). Removal of extracellular calcium attenuates not only the TRPV6-like conductance, but also the CaCC. Lanthanide block is time dependent such that the TRPV6-like component is inhibited first, followed by the CaCC. The putative CaCC blocker niflumic acid partially inhibits whole-cell currents, whereas La(3+) almost abolishes whole-cell currents in principal cells. Membrane potential measurements reveal an interplay between La(3+)-sensitive ion channels and those that are sensitive to the specific TMEM16A inhibitor tannic acid. In vivo perfusion of the cauda epididymal tubule shows a substantial rate of Ca(2+) reabsorption from the luminal side, which is dose-dependently suppressed by ruthenium red, a putative blocker of epithelial Ca(2+) channels and CaCC. Finally, we discover messenger RNA for both TRPV6 and TMEM16A in the rat epididymis and show that their proteins colocalize in the apical membrane of principal cells. Collectively, these data provide evidence for a coupling mechanism between TRPV6 and TMEM16A in principal cells that may play an important role in the regulation of calcium homeostasis in the epididymis.

Proliferation and enrichment of CD133(+) glioblastoma cancer stem cells on 3D chitosan-alginate scaffolds.

Emerging evidence implicates cancer stem cells (CSCs) as primary determinants of the clinical behavior of human cancers, representing an ideal target for next-generation anti-cancer therapies. However CSCs are difficult to propagate in vitro, severely limiting the study of CSC biology and drug development. Here we report that growing cells from glioblastoma (GBM) cell lines on three dimensional (3D) porous chitosan-alginate (CA) scaffolds dramatically promotes the proliferation and enrichment of cells possessing the hallmarks of CSCs. CA scaffold-grown cells were found more tumorigenic in nude mouse xenografts than cells grown from monolayers. Growing in CA scaffolds rapidly promoted expression of genes involved in the epithelial-to-mesenchymal transition that has been implicated in the genesis of CSCs. Our results indicate that CA scaffolds have utility as a simple and inexpensive means to cultivate CSCs in vitro in support of studies to understand CSC biology and develop more effective anti-cancer therapies.

A simple material model to generate epidermal and dermal layers in vitro for skin regeneration.

There is an urgent need for a rationally-designed, cellularized skin graft capable of reproducing the micro-environmental cues necessary to promote skin healing and regeneration. To address this need, we developed a composite scaffold, namely, CA/C-PEG, composing of a porous chitosan-alginate (CA) structure impregnated with a thermally reversible chitosan-poly(ethylene glycol) (C-PEG) gel to incorporate skin cells as a bi-layered skin equivalent. Fibroblasts were encapsulated in C-PEG to simulate the dermal layer while the keratinocytes were seeded on the top of CA/C-PEG composite scaffold to mimic the epidermal layer. The CA scaffold provided mechanical support for the C-PEG gel and the C-PEG gel physically segregated the keratinocytes from fibroblasts in the construct. Three different tissue culture micro-environments were tested: CA scaffolds without C-PEG cultured in cell culture medium without air-liquid interface (-gel-interface), CA scaffolds impregnated with C-PEG and cultured in cell culture medium without air-liquid interface (-gel-interface), and CA scaffolds impregnated with C-PEG cultured in cell culture medium with air-liquid interface (-gel- interface). We found that the presence of C-PEG increased the cellular proliferation rates of both keratinocytes and fibroblasts, and the air-liquid interface induced keratinocyte maturation. This CA/C-PEG composite scaffold design is able to recapitulate micro-environments relevant to skin tissue engineering, and may be a useful tool for future skin tissue engineering applications.

Effect of nano- and micro-scale topological features on alignment of muscle cells and commitment of myogenic differentiation.

Skeletal muscle injury can lead to severe motor deficits that adversely affect movement and quality of life. Current surgical treatments for skeletal muscle are hindered by the poor formation of organized myotube bundles at the wound site. Tissue-engineered skeletal muscle constructs to date have been unable to generate high degrees of myotube density and alignment. Generating a suitable in vitro tissue-engineered skeletal muscle construct requires the design of a scaffold that recapitulates the structural combination of nanoscale collagen fibrils and aligned microscale basal lamina tracks present in the native extracellular matrix (ECM). We hypothesized that a 3D aligned tubular porous scaffold containing aligned nanofibers inside the pores can mimic the native muscle tissue environment. We constructed a laminar section of the hypothesized scaffold with aligned chitosan-PCL nanofibers arranged co-axially with the aligned microscale chitosan scaffold bands to mimic the required myogenic environment. A 6-day study of C2C12 mouse myoblast cells cultured on this hybrid scaffold indicated that the nanofibers and scaffold bands in the scaffold played a synergetic role in directing cell orientation, interaction, migration and organization. Our results showed that aligned nanofibers mediated cell alignment and the aligned scaffold bands induced the formation of a more compact assembly of myotube cells as compared to various control substrates including chitosan films, nanofibers, and chitosan bands. The expression levels of both early and late-stage myogenic differentiation genes associated with myogenin and myosin heavy chain, respectively, were higher on the hybrid substrate than on control substrates. Our study suggests that the combination of nano and microscale topological features in the ECM can direct myogenic differentiation, and the hybrid material has the potential to improve the outcome of skeletal tissue engineering.