Macromolecular crowding in human tenocyte and skin fibroblast cultures: A comparative analysis.

Although human tenocytes and dermal fibroblasts have shown promise in tendon engineering, no tissue engineered medicine has been developed due to the prolonged ex vivo time required to develop an implantable device. Considering that macromolecular crowding has the potential to substantially accelerate the development of functional tissue facsimiles, herein we compared human tenocyte and dermal fibroblast behaviour under standard and macromolecular crowding conditions to inform future studies in tendon engineering. Basic cell function analysis made apparent the innocuousness of macromolecular crowding for both cell types. Gene expression analysis of the without macromolecular crowding groups revealed expression of tendon related molecules in human dermal fibroblasts and tenocytes. Protein electrophoresis and immunocytochemistry analyses showed significantly increased and similar deposition of collagen fibres by macromolecular crowding in the two cell types. Proteomics analysis demonstrated great similarities between human tenocyte and dermal fibroblast cultures, as well as the induction of haemostatic, anti-microbial and tissue-protective proteins by macromolecular crowding in both cell populations. Collectively, these data rationalise the use of either human dermal fibroblasts or tenocytes in combination with macromolecular crowding in tendon engineering.

Blood biomarker-based classification study for neurodegenerative diseases.

As the population ages, neurodegenerative diseases are becoming more prevalent, making it crucial to comprehend the underlying disease mechanisms and identify biomarkers to allow for early diagnosis and effective screening for clinical trials. Thanks to advancements in gene expression profiling, it is now possible to search for disease biomarkers on an unprecedented scale.Here we applied a selection of five machine learning (ML) approaches to identify blood-based biomarkers for Alzheimer's (AD) and Parkinson's disease (PD) with the application of multiple feature selection methods. Based on ROC AUC performance, one optimal random forest (RF) model was discovered for AD with 159 gene markers (ROC-AUC = 0.886), while one optimal RF model was discovered for PD (ROC-AUC = 0.743). Additionally, in comparison to traditional ML approaches, deep learning approaches were applied to evaluate their potential applications in future works. We demonstrated that convolutional neural networks perform consistently well across both the Alzheimer's (ROC AUC = 0.810) and Parkinson's (ROC AUC = 0.715) datasets, suggesting its potential in gene expression biomarker detection with increased tuning of their architecture.

Glial Cell Adhesion Molecule (GlialCAM) Determines Proliferative versus Invasive Cell States in Glioblastoma.

The malignant brain cancer glioblastoma (GBM) contains groups of highly invasive cells that drive tumor progression as well as recurrence after surgery and chemotherapy. The molecular mechanisms that enable these GBM cells to exit the primary mass and disperse throughout the brain remain largely unknown. Here we report using human tumor specimens and primary spheroids from male and female patients that glial cell adhesion molecule (GlialCAM), which has normal roles in brain astrocytes and is mutated in the developmental brain disorder megalencephalic leukoencephalopathy with subcortical cysts (MLC), is differentially expressed in subpopulations of GBM cells. High levels of GlialCAM promote cell-cell adhesion and a proliferative GBM cell state in the tumor core. In contrast, GBM cells with low levels of GlialCAM display diminished proliferation and enhanced invasion into the surrounding brain parenchyma. RNAi-mediated inhibition of GlialCAM expression leads to activation of proinvasive extracellular matrix adhesion and signaling pathways. Profiling GlialCAM-regulated genes combined with cross-referencing to single-cell transcriptomic datasets validates functional links among GlialCAM, Mlc1, and aquaporin-4 in the invasive cell state. Collectively, these results reveal an important adhesion and signaling axis comprised of GlialCAM and associated proteins including Mlc1 and aquaporin-4 that is critical for control of GBM cell proliferation and invasion status in the brain cancer microenvironment.SIGNIFICANCE STATEMENT Glioblastoma (GBM) contains heterogeneous populations of cells that coordinately drive proliferation and invasion. We have discovered that glial cell adhesion molecule (GlialCAM)/hepatocyte cell adhesion molecule (HepaCAM) is highly expressed in proliferative GBM cells within the tumor core. In contrast, GBM cells with low levels of GlialCAM robustly invade into surrounding brain tissue along blood vessels and white matter. Quantitative RNA sequencing identifies various GlialCAM-regulated genes with functions in cell-cell adhesion and signaling. These data reveal that GlialCAM and associated signaling partners, including Mlc1 and aquaporin-4, are key factors that determine proliferative and invasive cell states in GBM.

Complexities in attributing lead contamination to specific sources in an industrial area of Philadelphia, PA.

Globally, lead (Pb) contamination is one of the top ten chemical exposure issues affecting public health. The identification of specific Pb sources provides valuable information to determine assignment of liability for site cleanup, improve sampling plans and develop remedial strategies. This paper examines Pb concentrations and Pb isotopic data from samples collected at and near the site of a Pb paint production facility with a long operating history. Although high soil Pb concentrations were found at the site, Pb concentrations in surrounding neighborhoods did not simply decline with distance from the site. We evaluated soil concentrations and isotopic mixing lines to explore potential sources of Pb pollution. Three-isotope plots showed overlap of site samples and the surrounding neighborhood, consistent with pollution from the facility affecting offsite soils. A major challenge in separation of potential sources, however, is that the isotopic signatures of other potential Pb sources fall within the range of the soil data. The long operational site history, soil disturbances, the presence of nearby smelters, and other local and remote sources affect identification of lead sources. This analysis demonstrates that source attribution can be confounded by incomplete site and material sourcing information. An integrated approach that includes in-depth site characterization and an evaluation of historical activities (e.g., Pb ores used over time, amounts of Pb emitted by all area smelters, land use changes, and soil disturbances) is important for determining source attribution. This analysis provides insight into future site investigations where soil lead contamination has resulted from a long industrial history in an urban setting.

The alpha7 integrin subunit in astrocytes promotes endothelial blood-brain barrier integrity.

The blood-brain barrier (BBB) is a vascular endothelial cell boundary that partitions the circulation from the central nervous system to promote normal brain health. We have a limited understanding of how the BBB is formed during development and maintained in adulthood. We used quantitative transcriptional profiling to investigate whether specific adhesion molecules are involved in BBB functions, with an emphasis on understanding how astrocytes interact with endothelial cells. Our results reveal a striking enrichment of multiple genes encoding laminin subunits as well as the laminin receptor gene Itga7, which encodes the alpha7 integrin subunit, in astrocytes. Genetic ablation of Itga7 in mice led to aberrant BBB permeability and progressive neurological pathologies. Itga7-/- mice also showed a reduction in laminin protein expression in parenchymal basement membranes. Blood vessels in the Itga7-/- brain showed separation from surrounding astrocytes and had reduced expression of the tight junction proteins claudin 5 and ZO-1. We propose that the alpha7 integrin subunit in astrocytes via adhesion to laminins promotes endothelial cell junction integrity, all of which is required to properly form and maintain a functional BBB.

The Strigolactone Pathway Is a Target for Modifying Crop Shoot Architecture and Yield.

Due to their sessile nature, plants have developed the ability to adapt their architecture in response to their environment. Branching is an integral component of plant architecture, where hormonal signals tightly regulate bud outgrowth. Strigolactones (SLs), being a novel class of phytohormone, are known to play a key role in branching decisions, where they act as a negative regulator of bud outgrowth. They can achieve this by modulating polar auxin transport to interrupt auxin canalisation, and independently of auxin by acting directly within buds by promoting the key branching inhibitor TEOSINTE BRANCHED1. Buds will grow out in optimal conditions; however, when conditions are sub-optimal, SL levels increase to restrict branching. This can be a problem in agricultural applications, as reductions in branching can have deleterious effects on crop yield. Variations in promoter elements of key SL-related genes, such as IDEAL PLANT ARCHITECTURE1, have been identified to promote a phenotype with enhanced yield performance. In this review we highlight how this knowledge can be applied using new technologies to develop new genetic variants for improving crop shoot architecture and yield.

Factor structure of the modified Rivermead Post-concussion Symptoms Questionnaire (mRPQ): an exploratory analysis with healthy adult simulators.

OBJECTIVE: Persistent postconcussion symptoms (PPCS) are challenging to diagnose. An improved diagnostic process could consider typical and atypical postconcussion symptoms. This study examined the structure of a modified Rivermead Post-concussion Symptoms Questionnaire (mRPQ) with both symptom types. METHOD: 298 adult volunteers were randomized into groups: honest responders, mild traumatic brain injury (mTBI) simulators (MS), and biased mTBI simulators (BMS). Both mTBI simulating groups were coached about mTBI and primed about the simulation context (compensation evaluation). The BMS group was also encouraged to bias (exaggerate) symptoms. The participants completed an online battery of tests, including the mRPQ. RESULTS: An exploratory factor analysis of the mRPQ (full sample) revealed a three-factor solution, including a separate dimension for atypical symptoms (all item loadings >0.45, ~4% of explained variance). The overall and group analyses of the standard RPQ items (typical symptoms) found a one- or two-factor solution, as did the analyses of atypical symptoms. CONCLUSIONS: Consistent with prior RPQ research, a unidimensional or bifactor structure was measurable from standard RPQ symptoms. Whilst this study did not find support for domain-level symptom scores for either typical or atypical symptoms, the findings support the use of an overall atypical symptoms score.

Validating the modified Rivermead Post-concussion Symptoms Questionnaire (mRPQ).

OBJECTIVE: Response distortions in the reporting of postconcussion symptoms can occur for many reasons. The Rivermead Post-concussion Symptoms Questionnaire (RPQ) was recently modified to include an embedded symptom validity indicator to test for atypical symptoms. The present study used a simulation design to investigate the psychometric properties of the modified RPQ (mRPQ). METHOD: 298 adult volunteers were randomised into three groups: honest responding (Controls, C) who reported actual, current symptoms; mild traumatic brain injury (mTBI) simulators (MS) who role played being injured, and; biased mTBI simulators (BMS) who role played being injured and were asked to bias (exaggerate) their response. The MS and BMS participants received instructions to support the simulation. All participants completed the mRPQ and a modified Neurobehavioral Symptom Inventory (mNSI). RESULTS: A 2 × 3 mixed ANOVA with one within-group variable (Symptom type: Standard or Atypical) and one between-group variable (Instruction type: C, MS, BMS) found a significant two-way interaction (p < .05, ηp2 = .08). CONCLUSIONS: The BMS group had score elevations for both standard and atypical postconcussion symptoms; therefore, both symptom types should be considered when evaluating for biased responding. The mRPQ has promising psychometric properties and should be further developed.

Longitudinal assessment of listening skills in UK infants with hearing aids using the LittlEARS® auditory questionnaire.

OBJECTIVE: To investigate listening skills in infant hearing aid users using the LittlEARS® Auditory Questionnaire (LEAQ). DESIGN: Caregivers completed the LEAQ, and hearing aid data logging was recorded, at infant age 3-7 months and 7-21 months. STUDY SAMPLE: Seventy infant hearing aid users with permanent bilateral hearing loss, no developmental comorbidities, aged 3-7 months at first visit. RESULTS: Infants with mild and moderate losses tended to have scores within the normative range at the early time point (88%), but 29% were below the normative range when older. Thirty percent of infants with severe hearing loss were outside the normative range at the early time point and 60% outside the normative range when older. Infants with profound loss were almost always (95%) outside the normative range. At the later time point, and for infants with severe-to-profound loss, low LEAQ scores were associated with fewer daily hours hearing aid use. Scores were poorer than previous reports in the literature for infant hearing aid users. CONCLUSIONS: This study provides further knowledge on infant listening performance and hearing aid use over time that can be used to guide management of individual cases and to develop and audit service quality improvements.

A review of causal discovery methods for molecular network analysis.

BACKGROUND: With the increasing availability and size of multi-omics datasets, investigating the casual relationships between molecular phenotypes has become an important aspect of exploring underlying biology andgenetics. There are an increasing number of methodlogies that have been developed and applied to moleular networks to investigate these causal interactions. METHODS: We have introduced and reviewed the available methods for building large-scale causal molecular networks that have been developed and applied in the past decade. RESULTS: In this review we have identified and summarized the existing methods for infering causality in large-scale causal molecular networks, and discussed important factors that will need to be considered in future research in this area. CONCLUSION: Existing methods to infering causal molecular networks have their own strengths and limitations so there is no one best approach, and it is instead down to the discretion of the researcher. This review also to discusses some of the current limitations to biological interpretation of these networks, and important factors to consider for future studies on molecular networks.

Use of a synthetic biodegradable temporising matrix after necrotising fasciitis infection of the thigh.

Biodegradable temporising matrix (BTM NovoSorb; PolyNovo, Melbourne, Australia) is a novel synthetic polyurethane dermal substitute. Licensed in Europe in 2020, it was developed primarily for reconstruction of infected wounds. We present a case of a healthy man in his 60s with necrotising fasciitis of his left thigh. His medical history was significant for recurrent left thigh liposarcoma that was treated years earlier with surgical excision and adjuvant radiotherapy. The affected area was within the previously irradiated tissue, debrided down to fascia and dressed with a vacuum-assisted closure to help regenerate the wound bed. Reconstruction options were limited by having a circumferential thigh defect that was infected. Following the use of NovoSorb BTM, the area was dressed with Acticoat Flex 7 antimicrobial barrier dressing for 5 weeks. Patient mobilisation was permitted. The material integrated very well and formed a soft, pliable healthy dermal layer that was autografted with split thickness skin grafts. This resulted in durable cover of the thigh with good aesthetic contour and minimal contracture.

Examining the Content and Outcomes of Training in Dysphagia and Mealtime Management: A Systematic Review Informing Co-Design of New Training.

PURPOSE: Dysphagia (swallowing difficulty) impacts physical health, quality of life, and mealtime enjoyment. Staff who provide mealtime assistance to people with dysphagia require adequate training to help ensure that the mealtimes are safe and enjoyable. This systematic review examined literature relating to training in dysphagia (e.g., recognizing signs and symptoms) and mealtime assistance, its components, and benefits for people with dysphagia. METHOD: In July 2020, five scientific databases were searched for papers meeting the inclusion criteria relating to mealtime assistance training. The quality of the studies was evaluated using the Quality Assessment Tool for Studies of Diverse Design, with scores ranging from 38.1% to 83.3%. We completed a qualitative synthesis using the data extracted from the included studies. RESULTS: Twenty-four studies met the inclusion criteria. Participants in these studies benefited from both group training and one-on-one training. Training programs had many formats including computer-based, face-to-face, individual training, and group training. Each included study demonstrated some level of positive impact to the learners, such as improved knowledge and skills in mealtime management for people with dysphagia. No studies reported negative outcomes. Training duration ranged from 30 min to 5 days. CONCLUSIONS: The benefits of different components of mealtime training (e.g., group training, or face-to-face training) for mealtime assistance for people with dysphagia were reviewed. Further research is needed to compare the effectiveness of different training formats, involving not only the assistant but also people with dysphagia as both trainers and trainees, and determine the health outcomes of training programs for people with dysphagia.

Malignant epithelioid hemangioendothelioma of the ear.

Skin cancer is the most common cancer in Ireland. Our patient presented for removal of a cutaneous lesion on his ear. The histopathological diagnosis was malignant epithelioid hemangioendothelioma. It is very rare for this to present primarily as a cutaneous lesion. Here, we discuss the management of this patient and the surveillance he required. It is important to consider alternative histological diagnoses in patients presenting with cutaneous lesions and how this affects management and prognosis.

Shedding Light on SARS-CoV-2, COVID-19, COVID-19 Vaccination, and Auditory Symptoms: Causality or Spurious Conjunction?

There are reports of associations between SARS-CoV2, COVID-19, COVID-19 vaccines, and auditory symptoms (hearing difficulty, tinnitus). However, most studies have relied on self-report and lack baseline and/or non-COVID control groups. This makes it problematic to differentiate if symptoms are associated with SARS-CoV2, COVID-19, the vaccine, psychosocial factors or recall bias. In this study, we differentiate these by comparing hearing and tinnitus survey data collected pre- and during the pandemic. The survey conducted during the pandemic asked about the onset and change in three types of symptom. Type One-known association (loss of smell, memory/concentration issues, persistent fatigue), Type Two-indeterminate association (auditory symptoms), and Type Three-no established association with COVID-19 (toothache). We hypothesized that if auditory symptoms are directly associated with COVID-19, their onset and change would be similar to Type One symptoms, but if indirectly associated (reflecting psychosocial factors and/or recall bias) would be more similar to Type Three symptoms. Of the 6,881 individuals who responded, 6% reported confirmed COVID-19 (positive test), 11% probably had COVID-19, and 83% reported no COVID-19. Those with confirmed or probable COVID-19 more commonly reported new and/or worsened auditory symptoms than those not reporting COVID-19. However, this does not imply causality because: (1) new auditory symptoms coincided with COVID-19 illness among just 1/3 of those with confirmed or probable COVID-19, and another 1/3 said their symptoms started before the pandemic-despite reporting no symptoms in the pre-pandemic survey. (2) >60% of individuals who had COVID-19 said it had affected their Type 3 symptoms, despite a lack of evidence linking the two. (3) Those with confirmed COVID-19 reported more Type 1 symptoms, but reporting of Type 2 and Type 3 symptoms did not differ between those with confirmed COVID-19 and those without COVID-19, while those who probably had COVID-19 most commonly reported these symptom types. Despite more reports of auditory symptoms in confirmed or probable COVID-19, there is inconsistent reporting, recall bias, and possible nocebo effects. Studies that include appropriate control groups and use audiometric measures in addition to self-report to investigate change in auditory symptoms relative to pre-COVID-19 are urgently needed.

GMP-Compliant Production of Autologous Adipose-Derived Stromal Cells in the NANT 001 Closed Automated Bioreactor.

In recent years mesenchymal stromal cells (MSCs) have received a great deal of interest for the treatment of major diseases, but clinical translation and market authorization have been slow. This has been due in part to a lack of standardization in cell manufacturing protocols, as well as a lack of biologically meaningful cell characterization tools and release assays. Cell production strategies to date have involved complex manual processing in an open environment which is costly, inefficient and poses risks of contamination. The NANT 001 bioreactor has been developed for the automated production of small to medium cell batches for autologous use. This is a closed, benchtop system which automatically performs several processes including cell seeding, media change, real-time monitoring of temperature, pH, cell confluence and cell detachment. Here we describe a validation of the bioreactor in an environment compliant with current good manufacturing practice (cGMP) to confirm its utility in replacing standardized manual processing. Stromal vascular fraction (SVF) was isolated from lipoaspirate material obtained from healthy donors. SVF cells were seeded in the bioreactor. Cell processing was performed automatically and cell harvesting was triggered by computerized analysis of images captured by a travelling microscope positioned beneath the cell culture flask. For comparison, the same protocol was performed in parallel using manual methods. Critical quality attributes (CQA) assessed for cells from each process included cell yield, viability, surface immunophenotype, differentiation propensity, microbial sterility and endotoxin contamination. Cell yields from the bioreactor cultures were comparable in the manual and automated cultures and viability was >90% for both. Expression of surface markers were consistent with standards for adipose-derived stromal cell (ASC) phenotype. ASCs expanded in both automated and manual processes were capable of adipogenic and osteogenic differentiation. Supernatants from all cultures tested negative for microbial and endotoxin contamination. Analysis of labor commitment indicated considerable economic advantage in the automated system in terms of operator, quality control, product release and management personnel. These data demonstrate that the NANT 001 bioreactor represents an effective option for small to medium scale, automated, closed expansion of ASCs from SVF and produces cell products with CQA equivalent to manual processes.

A retrospective cohort study of cutaneous squamous cell carcinoma of the scalp: features of disease and influence of sociodemographic factors on outcomes.

BACKGROUND: Cutaneous squamous cell carcinoma (SCC) is an increasingly prevalent and potentially fatal disease with considerable implications if not recognized early and treated promptly. Several disease features contribute to a higher risk profile and adverse outcomes in affected patients. AIMS: Given the clinical observation that elderly males from rural communities often present with large SCCs of the scalp, we sought to investigate and describe features of disease and sociodemographic factors from a cohort of patients with scalp SCCs. METHODS: Histology reports of scalp primary SCCs were retrospectively assessed. Disease and demographic features were recorded. Descriptive statistics were generated, and statistical analyses (Fisher's exact, Mann-Whitney U and Spearman's rank test) were utilized to examine relationships between high-risk disease features and sociodemographic features. RESULTS: Ninety-three occurrences of scalp SCC in 61 patients were assessed. The average age at presentation was 78.81 years. Males were predominantly affected at a 14:1 ratio. Half of all tumours were greater than 2 cm (47/93 (50.54%)). The geographical distance from treatment was significantly associated with larger tumours at presentation. (rs = .34 P = 0.002). Recurrence and metastasis rates were determined amongst 188 patients with a primary scalp SCC, and low rates were observed (2.66% and 2.13%, respectively). CONCLUSIONS: Elderly males are inordinately affected by scalp SCC compared to females. Those living further from care exhibited larger tumours at presentation. Data from this study characterize features of SCC of the scalp and provide evidence to suggest that rural isolation may act as a mediator of high-risk presentation and larger tumour size.

Skin cancer margin detection using nanosensitive optical coherence tomography and a comparative study with confocal microscopy.

Excision biopsy and histology represent the gold standard for morphological investigation of the skin, in particular for cancer diagnostics. Nevertheless, a biopsy may alter the original morphology, usually requires several weeks for results, is non-repeatable on the same site and always requires an iatrogenic trauma. Hence, diagnosis and clinical management of diseases may be substantially improved by new non-invasive imaging techniques. Optical Coherence Tomography (OCT) is a non-invasive depth-resolved optical imaging modality based on low coherence interferometry that enables high-resolution, cross-sectional imaging in biological tissues and it can be used to obtain both structural and functional information. Beyond the resolution limit, it is not possible to detect structural and functional information using conventional OCT. In this paper, we present a recently developed technique, nanosensitive OCT (nsOCT), improved using broadband supercontinuum laser, and demonstrate nanoscale sensitivity to structural changes within ex vivo human skin tissue. The extended spectral bandwidth permitted access to a wider distribution of spatial frequencies and improved the dynamic range of the nsOCT. Firstly, we demonstrate numerical and experimental detection of a few nanometers structural difference using the nsOCT method from single B-scan images of phantoms with sub-micron periodic structures, acting like Bragg gratings, along the depth. Secondly, our study shows that nsOCT can distinguish nanoscale structural changes at the skin cancer margin from the healthy region in en face images at clinically relevant depths. Finally, we compare the nsOCT en face image with a high-resolution confocal microscopy image to confirm the structural differences between the healthy and lesional/cancerous regions, allowing the detection of the skin cancer margin.

Interpreting wide-band neural activity using convolutional neural networks.

Rapid progress in technologies such as calcium imaging and electrophysiology has seen a dramatic increase in the size and extent of neural recordings. Even so, interpretation of this data requires considerable knowledge about the nature of the representation and often depends on manual operations. Decoding provides a means to infer the information content of such recordings but typically requires highly processed data and prior knowledge of the encoding scheme. Here, we developed a deep-learning framework able to decode sensory and behavioral variables directly from wide-band neural data. The network requires little user input and generalizes across stimuli, behaviors, brain regions, and recording techniques. Once trained, it can be analyzed to determine elements of the neural code that are informative about a given variable. We validated this approach using electrophysiological and calcium-imaging data from rodent auditory cortex and hippocampus as well as human electrocorticography (ECoG) data. We show successful decoding of finger movement, auditory stimuli, and spatial behaviors - including a novel representation of head direction - from raw neural activity.

Novel steroid infiltration technique of the carpal tunnel.

This report describes a novel technique of steroid infiltration of the wrist to treat symptomatic carpal tunnel syndrome. Our approach potentially reduces direct trauma to the median nerve when compared with current conventional techniques. The use of a cannula allows infiltration directly into the carpal tunnel and advancement of the blunt tip minimises the risk of sharp trauma to the median nerve and adjacent tendons. This avoids the unpleasant, shooting pain frequently experienced by patients using traditional needle infiltration. We anticipate this would be of particular benefit in reducing pain associated with the procedure.