Minimally-invasive Measurement Research Articles

Neurodegenerative fluid biomarkers are enriched in human cervical lymph nodes.

In animal models, brain neurodegeneration biomarkers drain into cervical lymph nodes (CLNs), and this drainage function is reduced with ageing. If this occurred in humans, CLNs may provide a readily accessible measure of this aspect of protein clearance. We tested this hypothesis in people using ultrasound-guided fine needle aspiration (FNA). We measured amyloid-beta 40 and 42, phospho-Tau-181, glial-fibrillary-acidic-protein, and neurofilament-light using single molecule array in CLN aspirates and plasma from: i) a discovery cohort of 25 autoimmune patients, and from ii) plasma, CLNs and capillary blood in four healthy volunteers, an optimisation cohort. FNA was well-tolerated by all participants. In both cohorts, all biomarkers were detected in all plasma and CLN samples, other than neurofilament-light (8/17 of discovery cohort). CLN biomarker concentrations were significantly greater than plasma concentrations for all except neurofilament-light, most markedly for phospho-Tau-181 (266-fold; P<0.02), whose CLN concentrations decreased with age (Spearman r=-0.66, P=0.001). This study presents the first evidence that neurodegenerative biomarkers are detectable in human CLNs. Raised CLN:plasma biomarker ratios suggest their concentration in CLNs may offer a distinct compartment for minimally-invasive measurement of brain clearance and lymphatic drainage, with potential applicability to study of ageing and future clinical trials.

Minimally-Invasive Measurement of Small Intestinal and Stool Microbiomes in Humans Reveals Distinct Community Structure and Composition: A Novel Tool for Dietary Studies

Minimally-Invasive Measurement of Small Intestinal and Stool Microbiomes in Humans Reveals Distinct Community Structure and Composition: A Novel Tool for Dietary Studies

Interleukin-17A blockade with secukinumab results in decreased neutrophil infiltration in psoriasis: minimally-invasive measurement by tape stripping

Psoriasis is a well characterized interleukin (IL)-17A-driven skin disease with neutrophil infiltration and epidermal hyperkeratosis. Several biomarkers, most prominently β-defensin-2 (BD-2), have been identified using local andsystemic invasive measurements as responsive markers of IL-17A-driven skin pathology. We sought to determinewhether measurements of epidermal proteins by tape stripping could offer a minimally-invasive method to assess treatment responses. We compared the expression of 170 proteins in the epidermis (tape stripping) and dermis (open flowmicroperfusion) of 8 psoriatic subjects before and after administration of a single dose of subcutaneous (s.c.) antiIL-17A mAb secukinumab. Proteomic analyses of tape strips revealed a &gt;3-fold decrease in 32 epidermal and inflammatory cell proteins in response to secukinumab. The epidermal proteins with the largest (&gt;10-fold) decreases were: matrix metalloproteinase-8 (MMP-8, 15.68-fold, p&lt;0.05); myeloperoxidase (MPO, 14.72-fold, p&lt;0.005); IL-8 (11.93-fold, p&lt;0.05); MMP-9 (10.81-fold, p&lt;0.005); and IL-1β (10.35-fold, p&lt;0.05). For these proteins, greater-fold protein changes were detected in the epidermis compared to dermis. Immunohistochemical analysis confirmed that neutrophils are the predominant cell type in psoriatic skin lesions that express MPO, MMP-8 and MMP-9, and that secukinumab treatment dramatically decreases neutrophil accumulation. Thus, tape stripping may be used to assess epidermal neutrophils, and protein biomarker responses to anti-IL-17A therapy in psoriasis.

An Activity-Based Nanosensor for Minimally-Invasive Measurement of Protease Activity in Traumatic Brain Injury.

Current screening and diagnostic tools for traumatic brain injury (TBI) have limitations in sensitivity and prognostication. Aberrant protease activity is a central process that drives disease progression in TBI and is associated with worsened prognosis; thus direct measurements of protease activity could provide more diagnostic information. In this study, a nanosensor is engineered to release a measurable signal into the blood and urine in response to activity from the TBI-associated protease calpain. Readouts from the nanosensor were designed to be compatible with ELISA and lateral flow assays, clinically-relevant assay modalities. In a mouse model of TBI, the nanosensor sensitivity is enhanced when ligands that target hyaluronic acid are added. In evaluation of mice with mild or severe injuries, the nanosensor identifies mild TBI with a higher sensitivity than the biomarker GFAP. This nanosensor technology allows for measurement of TBI-associated proteases without the need to directly access brain tissue, and has the potential to complement existing TBI diagnostic tools.

Non-Invasive and Minimally-Invasive Cerebral Autoregulation Assessment: A Narrative Review of Techniques and Implications for Clinical Research.

The process of cerebral vessels regulating constant cerebral blood flow over a wide range of systemic arterial pressures is termed cerebral autoregulation (CA). Static and dynamic autoregulation are two types of CA measurement techniques, with the main difference between these measures relating to the time scale used. Static autoregulation looks at the long-term change in blood pressures, while dynamic autoregulation looks at the immediate change. Techniques that provide regularly updating measures are referred to as continuous, whereas intermittent techniques take a single at point in time. However, a technique being continuous or intermittent is not implied by if the technique measures autoregulation statically or dynamically. This narrative review outlines technical aspects of non-invasive and minimally-invasive modalities along with providing details on the non-invasive and minimally-invasive measurement techniques used for CA assessment. These non-invasive techniques include neuroimaging methods, transcranial Doppler, and near-infrared spectroscopy while the minimally-invasive techniques include positron emission tomography along with magnetic resonance imaging and radiography methods. Further, the advantages and limitations are discussed along with how these methods are used to assess CA. At the end, the clinical considerations regarding these various techniques are highlighted.

Electrical Bio-Impedance Proximity Sensing for Vitreo-Retinal Micro-Surgery

Retinal vein occlusion is a widespread eye vascular disease leading to vision loss because of clots obstructing a retinal vessel. Recent research investigated retinal vein cannulation, a promising treatment in which a microneedle injects a thrombolytic agent inside the clotted vessel. During this surgical procedure, the surgeon only relies on a microscope, looking through the patient eye's lens. Such visual feedback gives poor depth perception, with a risk to inject the agent underneath the retina. Such situation endangers the patient's eyesight and is to be avoided. This letter explores bio-impedance as a means to estimate the proximity between an insulated electrode and a retinal vessel. Experiments on five ex-vivo pig eyes showed that the impedance phase decreases as the electrode tip approaches the retinal vessel. Specific patterns in the impedance magnitude were detected. A dedicated algorithm showed 98 $\%$ sensitivity and 100 $\%$ specificity at a distance of 775 ± 275 μm away from the retinal vessels. Knowledge of the proximity can help the surgeon prevent damaging fragile retinal structures during retinal surgeries such as cannulation and scar tissue peeling. A minimally-invasive measurement principle of this technique is demonstrated on an ex-vivo pig head to show the feasibility of such technique in more realistic conditions.

Maternal salivary cortisone to cortisol ratio in late pregnancy: An improved method for predicting offspring birth weight

Overexposure to maternal cortisol in utero has been associated with lower birth weight of offspring. In order to regulate fetal exposure to this hormone, placental expression of 11-β-hydroxysteroid dehydrogenase-2, an enzyme that converts active cortisol to inactive cortisone, increases across pregnancy. Because of this increase in 11-βHSD2 activity, measuring maternal cortisol in isolation may not reflect actual fetal exposure to the hormone. Previous work by Hellgren et al. (2016) has shown that maternal serum cortisone:cortisol ratio was a better predictor of offspring birth weight than cortisol measured in isolation. This paper sought to replicate these results when examining maternal salivary cortisone:cortisol ratio. Data come from 55 pregnant women from Auckland, New Zealand. Cortisol and cortisone were measured in saliva samples collected at waking and prior to going to sleep on two consecutive weekdays between 34 and 36 weeks of gestation. We found that salivary cortisol and cortisone followed the expected diurnal rhythm and that cortisone was higher than cortisol at both times of day. Maternal bedtime cortisone:cortisol ratio was significantly and inversely related to offspring birth weight. However, waking and bedtime cortisol, as well as waking cortisone:cortisol ratio, were unrelated to birth weight. These results show that maternal salivary cortisone:cortisol ratio, like serum cortisone:cortisol ratio, is a more sensitive biomarker for predicting infant birth weight than cortisol measured in isolation. This ratio could be a valuable, minimally-invasive measurement for future studies interested in understanding the relationship between maternal HPA-axis function and offspring birth weight.

Microneedle-based minimally-invasive measurement of puncture resistance and fracture toughness of sclera.

The sclera provides the structural support of the eye and protects the intraocular contents. Since it covers a large portion of the eye surface and has relatively high permeability for most drugs, the sclera has been used as a major pathway for drug administration. Recently, microneedle (MN) technology has shown the possibility of highly local and minimally-invasive drug delivery to the eye by MN insertion through the sclera or the suprachoroidal space. Although ocular MN needs to be inserted through the sclera, there has been no systematic study to understand the mechanical properties of the sclera, which are important to design ocular MNs. In this study, we investigated a MN-based method to measure the puncture resistance and fracture toughness of the sclera. To reflect the conditions of MN insertion into the sclera, force-displacement curves obtained from MN-insertion tests were used to estimate the puncture resistance and fracture toughness of sclera tissue. To understand the effect of the insertion conditions, dependency of the mechanical properties on insertion speeds, pre-strain of the sclera, and MN sizes were analyzed and discussed. Measurement of mechanical property of soft biological tissue is challenging due to variations between tissue samples or lack of well-defined measurement techniques. Although non-invasive measurement techniques such as nano/micro indentation were employed to locally measure the elastic modulus of soft biological materials, mechanical properties such as puncture resistance or fracture toughness, which requires "invasive" measurement and is important for the application of "microneedles or hypodermic needles", has not been well studied. In this work, we report minimally-invasive measurement of puncture resistance and fracture toughness of sclera using a double MN insertion method. Parametric studies showed that use of MN proved to be advantageous because of minimally-invasive insertion into tissue as well as higher sensitivity to sub-tissue architecture during the measurement.

Minimally-invasive transepidermal potentiometry with microneedle salt bridge.

A commercial painless microneedle was filled with physiological saline agar, and this needle-based salt bridge was inserted into the skin (a piece of porcine skin and a flank skin of a live mouse) to make an electrical contact with its subepidermal region. The transepidermal potential (TEP), the potential difference between the skin surface and the subepidermal region, was measured using this inner electrode and a conventional agar electrode on the surface of the skin. Control of penetration depth of the inner electrode with a spacer and hydrophilic pretreatment with ozone plasma were found to be necessary for stable measurement. The TEP was reduced upon damages on the skin surface by tape stripping and acetone defatting, which indicated the fabricated needle electrode is useful for the minimally-invasive measurement of TEP and evaluation of skin barrier functions. Furthermore, we showed that the device integrating two electrodes into a single compact probe was useful to evaluate the local barrier functions and their mapping on a skin. This device could be a personal diagnostic tool in the fields of medicine and cosmetics in future.

BOLD fMRI in awake prairie voles: A platform for translational social and affective neuroscience

The advancement of neuroscience depends on continued improvement in methods and models. Here, we present novel techniques for the use of awake functional magnetic resonance imaging (fMRI) in the prairie vole (Microtus ochrogaster) — an important step forward in minimally-invasive measurement of neural activity in a non-traditional animal model. Imaging neural responses in prairie voles, a species studied for its propensity to form strong and selective social bonds, is expected to greatly advance our mechanistic understanding of complex social and affective processes. The use of ultra-high-field fMRI allows for recording changes in region-specific activity throughout the entire brain simultaneously and with high temporal and spatial resolutions. By imaging neural responses in awake animals, with minimal invasiveness, we are able to avoid the confound of anesthesia, broaden the scope of possible stimuli, and potentially make use of repeated scans from the same animals. These methods are made possible by the development of an annotated and segmented 3D vole brain atlas and software for image analysis. The use of these methods in the prairie vole provides an opportunity to broaden neuroscientific investigation of behavior via a comparative approach, which highlights the ethological relevance of pro-social behaviors shared between voles and humans, such as communal breeding, selective social bonds, social buffering of stress, and caregiving behaviors. Results using these methods show that fMRI in the prairie vole is capable of yielding robust blood oxygen level dependent (BOLD) signal changes in response to hypercapnic challenge (inhaled 5% CO2), region-specific physical challenge (unilateral whisker stimulation), and presentation of a set of novel odors. Complementary analyses of repeated restraint sessions in the imaging hardware suggest that voles do not require acclimation to this procedure. Taken together, awake vole fMRI represents a new arena of neurobiological study outside the realm of traditional rodent models.

The effect of consignment to broodmare Sales on physiological stress measured by faecal glucocorticoid metabolites in pregnant Thoroughbred mares

BackgroundValidation of a method for the minimally-invasive measurement of physiological stress will help understanding of risk factors that may contribute to stress-associated events including recrudescence of Equid herpesvirus (EHV), which is anecdotally associated with sales consignment of pregnant Thoroughbred mares. In this study we compared two similar groups of late-gestation Thoroughbred broodmares on the same farm: a consigned Sales group (N = 8) and a non-consigned Control group (N = 6). The Sales mares were separated from their paddock companions and grouped prior to their preparation for, transport to, and return from the sales venue. Both groups were monitored by sampling at regular intervals from 5 days prior to until 14 days after the sales date (D0) to measure physiological stress in terms of changes in faecal glucocorticoid metabolite (FGM) concentrations, and for event-related viral recrudescence via daily body temperature measurements and periodic nasal swabs for PCR analysis for EHV-1 and -4 DNA.ResultsIn both groups, FGM levels increased post-sales before returning to pre-sales levels. Specifically, FGM concentrations in the Sales mares were significantly higher on D + 3 and D + 10 than on D-4 and D-3 (F = 12.03, P < 0.0001, Post hoc: P = 0.0003 – 0.0008) and in the Control group FGM concentrations were higher on D + 10 than D-4 (F = 5.52, P = 0.004, Post hoc: P = 0.005). Interestingly, mean FGM levels in Control mares were significantly higher at 4 of the 5 sampling points (t = 5.64 – 2.25, p = 0.0001 – 0.044). Only one (Sales) mare showed PCR evidence of EHV-1 shedding.ConclusionsUsing FGM to measure physiological stress was supported by the increases observed in all mares after Sales consignment, including those not consigned to the sale. Monitoring FGM levels therefore represents an appropriate, minimally-invasive method for future studies to assess the contribution of physiological stress to EHV recrudescence in horses transported to sales or equestrian events.

Characterisation of periodically poled materials using nonlinear microscopy

Periodically poled crystalline materials are extremely attractive for processes such as second harmonic generation and optical parametric generation due to their very high conversion efficiency. For optimal performance, fabrication of poled regions with sub-micron tolerance is required. In this paper we introduce multi-photon laser scanning luminescence microscopy as a powerful minimally-invasive measurement technique which provides information about internal device structure with high spatial resolution that cannot be easily obtained with existing methods. A comparative study of confocal and multi-photon imaging of periodically poled crystalline materials is also performed.