The design and characterisation of sinusoidal toolpaths using sub-zero bioprinting of polyvinyl alcohol.

Sub-zero (°C) additive manufacturing (AM) systems present a promising solution for the fabrication of hydrogel structures with complex external geometry or a heterogeneous internal structure. Polyvinyl alcohol cryogels (PVA-C) are promising tissue-mimicking materials, with mechanical properties that can be designed to satisfy a wide variety of soft tissues. However, the design of more complex mechanical properties into additively manufactured PVA-C samples, which can be enabled using the toolpath, is a largely unstudied area. This research project will investigate the effect of toolpath variation on the elastic and viscoelastic properties of PVA-C samples fabricated using a sinusoidal toolpath. Samples were fabricated using parametric variation of a sinusoidal toolpath, whilst retaining the same overall cross-sectional area, using a sub-zero AM system. To mechanically characterise the samples, they were tested under tension in uniaxial ramp tests, and through dynamic mechanical analysis (DMA). The elastic and viscoelastic moduli of the samples are presented. No correlations between the parametric variation of the design and the Young's modulus were observed. Analysis of the data shows high intra-sample repeatability, demonstrated robust testing protocols, and variable inter-sample repeatability, indicating differences in the printability and consistency of fabrication between sample sets. DMA of the wavelength samples, show a frequency-dependent loss moduli. The storage modulus demonstrates frequency independence, and a large increase in magnitude as the sample increases to 3 wavelengths.

The orthotropic viscoelastic characterisation of sub-zero 3D-printed poly(vinyl alcohol) cryogel.

ABSTRACT: Poly(vinyl alcohol) cryogel (PVA) is a versatile biomaterial used to replicate the biomechanics of tissues. Additive manufacture (AM) at sub-zero (°C) temperatures enables the manufacture of PVA with complex geometry; however, the effect of processing parameters on the mechanical properties of PVA has not been evaluated. The aim of this study is to understand the impact of print nozzle diameter and orientation on the viscoelastic mechanical properties of PVA. Samples of sub-zero AM PVA, with different filament thicknesses, were tested under tension relative to the print direction, to calculate the storage and loss moduli. As the nozzle size was decreased, AM PVA exhibited more pronounced orthotropic properties; the smallest size showed a 33% decrease in storage moduli when tested perpendicular to the print direction, as opposed to parallel. This study has demonstrated the ability of sub-zero AM to tailor the orthotropic properties of PVA. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1557/s43580-021-00086-1.

The dynamic viscoelastic characterisation and magnetic resonance imaging of poly(vinyl alcohol) cryogel: Identifying new attributes and opportunities.

Poly(vinyl alcohol) (PVA) cryogel is a biocompatible, synthetic hydrogel, compatible with magnetic resonance (MR) imaging. It is widely used as a biomaterial in tissue scaffolds and mimics to test various diagnostic techniques. The aim of this study is to characterise the effect of varying PVA concentration, molecular weight (MW) and manufacturing protocol on the viscoelastic mechanical properties and MR T2 relaxation time. Further to this MR imaging (MRI) was investigated as a method to quantify material homogeneity. Cylindrical samples of PVA, of varying MW, concentration and number of freeze thaw cycles (FTCs), were manufactured. Dynamic mechanical analysis was performed to evaluate the storage and loss moduli between frequencies of 0.5 and 10 Hz. MR T2 relaxation maps were imaged using a 7 T MRI instrument. Storage and loss moduli were shown to increase with MW, concentration, or the number of FTCs; with storage modulus ranging from 55 kPa to 912 kPa and loss modulus ranging from 6 kPa to 103 kPa. MR T2 relaxation time was shown to increase linearly with PVA concentration. The qualitative and quantitative heterogeneity of the PVA sample were identified through MR T2 relaxation time maps. Excitingly, PVA demonstrated a composition-dependent casual correlation between the viscoelastic mechanical properties and MR T2 relaxation time. In conclusion, this research thoroughly characterised the viscoelastic mechanical properties of PVA to support its extensive use as a biomaterial, and demonstrated the use of MRI to non-invasively identify sample heterogeneity and to predict the composition-dependent viscoelastic properties of PVA.

Implementation of a psychodermatology clinic at a major health system in Detroit.

The association between psychiatric and dermatologic disorders has been well characterized in the present literature with estimates of up to 40% of dermatology patients having concomitant psychiatric problems that are often related to their skin condition. Here, we present our experience regarding the implementation of a psychodermatology clinic in Detroit, Michigan. The most commonly referred conditions were delusions of parasitosis, neurotic excoriations, and isotretinoin initiation for patients with a history of psychiatric conditions. Seventy-three percent of referred patients were female. By creating a monthly clinic for patients who are diagnosed with skin conditions and associated psychiatric disorders or psychological symptoms, we are able to meet the needs of these patients with a synergistic relationship between health care providers.

The Effect of Chemical Treatments on Lampenflora and a Collembola Indicator Species at a Popular Tour Cave in California, USA.

Growth of invasive, opportunistic plants (i.e. lampenflora) in popular tour caves is a significant concern for land managers worldwide. Numerous chemicals at various concentrations have been utilized to remove phototrophic lampenflora colonizing artificially lit surfaces within these caves; however formulations, effectiveness, and impacts appear anecdotal and temporally limited. At Crystal Cave, Sequoia National Park, California, we study lampenflora and cave springtail (Tomocerus celsus) response to a single 0.05 ml/cm2 dose of 1.0% sodium hypochlorite, 0.5% sodium hypochlorite, and 15.0% hydrogen peroxide compared to no treatment over the course of one year. Additionally, we explore potential food web impacts resulting from invasive lampenflora in naturally oligotrophic caves by utilizing stable isotope analysis of T. celsus found on and off lampenflora. Time-effect decay models indicate 1.0 and 0.5% sodium hypochlorite effectively eliminate lampenflora in 11 and 21 days, respectively, while lampenflora decay projections exceed 600 days with 15.0% hydrogen peroxide treatment. Repeat surveys of T. celsus indicate a negative response to 1.0% sodium hypochlorite (P = 0.02), and the probability of observing T. celsus was inversely related to the effectiveness of each treatment. Further, T. celsus had similar diets regardless of their lampenflora association (P = 0.92). We conclude that treatments of sodium hypochlorite at or below 0.5% achieve management goals with limited impacts to the presence or diet of a common cave-adapted indicator species.

The simulation of magnetic resonance elastography through atherosclerosis.

The clinical diagnosis of atherosclerosis via the measurement of stenosis size is widely acknowledged as an imperfect criterion. The vulnerability of an atherosclerotic plaque to rupture is associated with its mechanical properties. The potential to image these mechanical properties using magnetic resonance elastography (MRE) was investigated through synthetic datasets. An image of the steady state wave propagation, equivalent to the first harmonic, can be extracted directly from finite element analysis. Inversion of this displacement data yields a map of the shear modulus, known as an elastogram. The variation of plaque composition, stenosis size, Gaussian noise, filter thresholds and excitation frequency were explored. A decreasing mean shear modulus with an increasing lipid composition was identified through all stenosis sizes. However the inversion algorithm showed sensitivity to parameter variation leading to artefacts which disrupted both the elastograms and quantitative trends. As noise was increased up to a realistic level, the contrast was maintained between the fully fibrous and lipid plaques but lost between the interim compositions. Although incorporating a Butterworth filter improved the performance of the algorithm, restrictive filter thresholds resulted in a reduction of the sensitivity of the algorithm to composition and noise variation. Increasing the excitation frequency improved the techniques ability to image the magnitude of the shear modulus and identify a contrast between compositions. In conclusion, whilst the technique has the potential to image the shear modulus of atherosclerotic plaques, future research will require the integration of a heterogeneous inversion algorithm.

Absence of selenoprotein P but not selenocysteine lyase results in severe neurological dysfunction.

Dietary selenium restriction in mammals causes bodily selenium to be preferentially retained in the brain relative to other organs. Almost all the known selenoproteins are found in brain, where expression is facilitated by selenocysteine (Sec)-laden selenoprotein P. The brain also expresses selenocysteine lyase (Scly), an enzyme that putatively salvages Sec and recycles the selenium for selenoprotein translation. We compared mice with a genetic deletion of Scly to selenoprotein P (Sepp1) knockout mice for similarity of neurological impairments and whether dietary selenium modulates these parameters. We report that Scly knockout mice do not display neurological dysfunction comparable to Sepp1 knockout mice. Feeding a low-selenium diet to Scly knockout mice revealed a mild spatial learning deficit without disrupting motor coordination. Additionally, we report that the neurological phenotype caused by the absence of Sepp1 is exacerbated in male vs. female mice. These findings indicate that Sec recycling via Scly becomes limiting under selenium deficiency and suggest the presence of a complementary mechanism for processing Sec. Our studies illuminate the interaction between Sepp1 and Scly in the distribution and turnover of body and brain selenium and emphasize the consideration of sex differences when studying selenium and selenoproteins in vertebrate biology.

Reduction of plasma clot stability by a novel factor XIIIa inhibitor from the Giant Amazon Leech, Haementeria ghilianii.

The blood-sucking leech, Haementeria ghilianii, has evolved a number of agents that attenuate haemostasis. Recently we have isolated a potent inhibitor of factor XIIIa, tridegin, in the salivary glands which is almost certainly involved in feeding. Addition of purified natural tridegin to plasma, prior to clotting with thrombin, results in clots that deform more readily as adjudged by the greatly reduced development of the storage modulus on application of a shear force. The increase in the storage modulus in developing plasma clots is a slow process and continues for many hours. The effect of tridegin is particularly great when the clots are permitted to age in this way, demonstrating the role of factor XIIIa in the process. The IC50 for this inhibition is 138 ng/ml. Clots formed in the presence of tridegin are also lysed more rapidly in vitro by the leech's own fibrinolytic enzyme, hementin (time for 50% lysis, 16.0 +/- 0.8 h versus 22.3 +/- 2.0 h, P < 0.05). The synergy with which these agents act together may provide lessons for therapy of thrombosis in man.

Tridegin, a new peptidic inhibitor of factor XIIIa, from the blood-sucking leech Haementeria ghilianii.

1. Crude salivary gland extract of the giant Amazon leech, Haementeria ghilianii, contains an inhibitor of plasma factor XIIIa. 2. The inhibitory agent was purified to homogeneity by anion-exchange, cation-exchange, gel-filtration and reverse-phase chromatography to yield a single band on SDS/PAGE with an apparent molecular mass of 7.3 kDa. It has been named tridegin. 3. Micro-sequencing of proteolytic fragments showed tridegin to be a peptide of 66 amino acids. The sequence is unique with little similarity to other leech-derived proteins. 4. Inhibition of plasma factor XIIIa activity was confirmed by four independent methods: tridegin increased the solubility of fibrin clots in urea, inhibited ammonia produced from the incorporation of ethylamine into casein, inhibited the incorporation of 5'-(biotinamido)pentylamine into casein and prevented gamma-dimer formation in clotting fibrinogen. 5. The IC50 of tridegin (approx. 9.2 nM) is very close to the concentration of factor XIIIa used in the assay and in fact depends on its concentration. This is the most potent inhibitor of factor XIIIa yet described. 6. Tridegin also inhibits platelet factor XIIIa (factor XIIIAa) with a similar potency to that of the plasma enzyme. 7. Tridegin also inhibits tissue transglutaminase but with lower potency and independently of the enzyme concentration. 8. Tridegin appears to be specific for transglutaminases, since it has no effect on the coagulation times of human plasma, on thrombin or factor Xa. Moreover it has no effect on other thiol-containing enzymes and has no ability to digest fibrinogen or cleave the isopeptide substrate, L-gamma-glutamyl-4-nitroanilide.

Tridegin, a novel peptidic inhibitor of factor XIIIa from the leech, Haementeria ghilianii, enhances fibrinolysis in vitro.

Tridegin is a potent inhibitor of factor XIIIa from the leech, Haementeria ghilianii, which inhibits protein cross-linking. It modifies plasmin-mediated fibrin degradation as shown by the absence of D-dimer and approximately halves the time for fibrinolysis. Plasma clots formed in the presence of Tridegin lyse more rapidly when either streptokinase, tissue plasminogen activator or hementin is added 2 h after clot formation. The effect of Tridegin is markedly increased if clots are formed from platelet-rich plasma. Platelet-rich plasma clots are lysed much more slowly by the fibrinolytic enzymes used and if Tridegin is present, the rate of lysis returns almost to that of platelet-free clots. These studies indicate the important role of platelets in conferring resistance to commonly used fibrinolytic enzymes and suggest that protein cross-linking is an important step in this effect. Moreover they indicate that Tridegin, a small polypeptide, may have potential as an adjunct to thrombolytic therapy.

The in vitro characterization and biostability of 99mTc-dextran and its accumulation within the inflamed paws of adjuvant-induced arthritic rats.

The in vitro and in vivo stability in normal and adjuvant-induced arthritic rats of 99mTc-dextrans (10, 40 and 500 kDa) have been investigated. The circulation half-lives were molecular weight dependent, with 10 and 40 kDa fractions being cleared relatively rapidly due to their ability to cross the glomerular basement membrane. The 500 kDa dextran was eliminated more slowly although 79% had been removed from the circulation 4 h post injection which probably was due to its degradation by dextranases and subsequent glomerular excretion. Dextran accumulation by the RES was found to be similar for all molecular weight preparations with no significant differences found. The sequestration of the dextrans by tissues of the RES (liver, spleen and lung) was independent of clearance rate. No differences were seen between normal and arthritic groups. Accumulation of the polymers by inflamed paws greatly exceeded that of normal paws for the 10 kDa (5-fold) and 500 kDa (6-fold) although no differences were seen with the 40 kDa dextran.

Accumulation of polyvinylpyrrolidone within the inflamed paws of adjuvant-induced arthritic rats.

125I-Labelled polyvinylpyrrolidone ([125I]PVP) of a range of molecular weights (mol. wt 10, 40 and 360 kDa) was injected i.v. into adjuvant-induced arthritic and normal rats and the blood clearance and tissue distribution of the polymers determined. The half-life of PVP in the circulation increased with increasing mol. wt; 10, 40 and 360 kDa polymers had mean terminal half-lives of 2.2, 6.9 and 16.4 h, respectively. Tissue uptake was also found to be mol. wt dependent, the largest PVP molecule accumulating to a greater extent in the spleen, liver, lungs and paws in both normal and arthritic rats (P less than 0.01) than the two lower mol. wt polymers. Accumulation of the polymer in inflamed paws (g tissue)-1 greatly exceeded that of normal paws (P less than 0.01). This difference was particularly noticeable with 360 kDa PVP, where arthritic paws amassed 7 times more PVP than normal paws.

Prevention of herpesvirus infections in renal allograft recipients by low-dose oral acyclovir.

Forty patients with serum antibody against herpes simplex virus (HSV) were enrolled in a randomized, placebo-controlled, double-blind investigation of acyclovir given orally in a low dosage as prophylaxis against recurrent HSV infection after renal transplantation. During 30 postoperative days of medication, 14 of 21 placebo-treated and one of 19 acyclovir-treated patient(s) developed reactivation of HSV infection. Eleven of the former, but not the latter, had herpetic lesions. The protection against active infection with HSV during the period of prophylaxis with acyclovir is statistically highly significant. From 30 to 90 days after transplantation when no antiviral medicine was given, 60% (3/5) of the remaining placebo recipients and 44% (7/16) of the acyclovir patients developed active HSV infections. Herpetic lesions occurred in two of three and two of seven of infected people in the respective groups. No adverse effects of the drug were observed. The results show that HSV infections in immunosuppressed renal allograft recipients can be safely prevented, deferred, and ameliorated by an initial period of prophylaxis with a low dose of oral acyclovir.