Tailored Cyclodextrin Pore Blocker Protects Mammalian Cells from Clostridium difficile Binary Toxin CDT

Some Clostridium difficile strains produce, in addition to toxins A and B, the binary toxin Clostridium difficile transferase (CDT), which ADP-ribosylates actin and may contribute to the hypervirulence of these strains. The separate binding and translocation component CDTb mediates transport of the enzyme component CDTa into mammalian target cells. CDTb binds to its receptor on the cell surface, CDTa assembles and CDTb/CDTa complexes are internalised. In acidic endosomes, CDTb mediates the delivery of CDTa into the cytosol, most likely by forming a translocation pore in endosomal membranes. We demonstrate that a seven-fold symmetrical positively charged β-cyclodextrin derivative, per-6-S-(3-aminomethyl)benzylthio-β-cyclodextrin, which was developed earlier as a potent inhibitor of the translocation pores of related binary toxins of Bacillus anthracis, Clostridium botulinum and Clostridium perfringens, protects cells from intoxication with CDT. The pore blocker did not interfere with the CDTa-catalyzed ADP-ribosylation of actin or toxin binding to Vero cells but inhibited the pH-dependent membrane translocation of CDTa into the cytosol. In conclusion, the cationic β-cyclodextrin could serve as the lead compound in a development of novel pharmacological strategies against the CDT-producing strains of C. difficile.     

The Effects of Matrigel® on the Survival and Differentiation of a Human Neural Progenitor Dissociated Sphere Culture

We have previously developed an in vitro organotypic culture setting in order to investigate the performance of cellular substrates transplanted to the auditory nervous system. We have utilized this system to predict the efficacy of human neural progenitor cells (HNPCs) in transplantation to the auditory nerve to facilitate regeneration of sensory auditory nerve structures in vivo and in vitro. To optimize the growth and differentiation of HNPCs we have introduced an expansion of our in vitro system, exploring the impact of a growth factor-altered microenvironment. Here, we seeded HNPCs as a dissociated sphere culture on a hydrogel matrix coating (Matrigel®). We evaluated the performance of HNPCs by studying their survival, differentiation, and their axon-forming capacity. In identical culture conditions, we found that the overall survival rate of HNPCs on Matrigel coated surfaces was better than that on surfaces that were not coated with Matrigel. Furthermore, cells on Matrigel differentiated into neuronal cells to a far greater extent leading to strong synaptic marker signatures. Overall, our findings show that the present Matrigel matrix setting offers an experimental environment for the HNPCs to grow where these cells show novel and promising phenotypic characteristics suitable for further in vivo transplantation to the auditory nerve. Anat Rec, 303:441–450, 2020. © 2019 American Association for Anatomy     

Inkjet assisted fabrication of planar biocompatible memristors

In this study we address a novel design of a planar memristor and investigate its biocompatibility. An experimental prototype of the proposed memristor assembly has been manufactured using a hybrid nanofabrication method, combining sputtering of electrodes, patterning the insulating trenches, and filling them with a memristive substance. To pattern the insulating trenches, we have examined two nanofabrication techniques employing either a focused ion beam or a cantilever tip of an atomic force microscope. Inkjet printing has been used to fill the trenches with the functional titania ink. The experimental prototypes have qualitatively demonstrated memristive current–voltage behavior, as well as high biocompatibility.

A planar memristor was fabricated by a hybrid method combining AFM patterning and inkjet printing.     

Long term follow up of through-the-scope balloon dilation as compared to strictureplasty and bowel resection of intestinal strictures in crohn’s disease

Background & aims: Ileo-colonic strictures are common complication of Crohn’s disease (CD), and may result in repeated endoscopic or surgical therapy with a risk of further complications, such as perforation or short bowel syndrome. Strictures develop as a consequence of tissue remodelling and fibrosis due to chronic inflammation. This study compares the outcome of CD patients undergoing primarily endoscopic treatment with those undergoing surgery at an university hospital. Methods: In this study we retrospectively included 88 CD patients with intestinal strictures (37 males, 51 females, mean age 40 years, range 19-65 years) of both our medical and our surgical department, who underwent either surgical or endoscopic therapy between January 2002 and January 2006 with prospective, controlled follow-up, extended till January 2010 (mean follow-up period: 5 years; range 4-8 years). The primary end-point was operation- and symptom-free time. Patients were primarily divided into four groups: only surgical therapy, only endoscopic therapy, endoscopy with subsequent surgery, and initial surgical therapy followed by endoscopic dilations. Results: 53% of all patients remained surgery-free with mean follow-up of 49 months; a single endoscopic dilation was sufficient enough in 9 patients to achieve a surgery-free time of 51 months, other patients required up to 5 dilations. The average interval between first and second dilation was 6.5 months, between second and third 10.5 months. In the group of patients with only endoscopic therapy, surgery- and symptom-free time was shorter, as compared to the group of only surgical therapy. We found that stenoses in the surgical group with an average length of 6.5 cm were as expected longer, as compared to the endoscopic group (3 cm, ranging from 2-4 cm). The surgery-free time was 49 months (42-71 months, P = 0.723) with a symptom-free time of 12 months (4.5-46 months, P = 0.921). In the group of only surgically treated patients, 68.4% of the patients had only one stenosis, 18.4% had 2-3 stenoses and 13.2% more than 3 stenoses. In all patients the surgery- and symptoms- free time was 69 months (57-83 months, P = 0.850 and 0.908). The other two groups showed similar results. We found no significant effect of characteristic of stenosis (length, inflammation, the number of stenoses), injection of prednisolone, disease activity at the time of dilation and medication at the time of dilation on the long-term outcome. Importantly, the success of symptom free time correlated with the diameter of the balloon. Conclusions: Endoscopic dilation should be considered as a first-line therapy for short, accessible, fibrotic strictures. Careful patient selection and proper diagnostic imaging pre-procedure are essential requirements for safe and successful treatment. The balloon diameter seems to correlate positively with the long term outcome of dilation. However, at ever shorter intervals between endoscopic interventions, surgery should be discussed as an option for further treatment.     

The influence of the sacrificial agent nature on transformations of the Zn(OH)2/Cd0.3Zn0.7S photocatalyst during hydrogen production under visible light

Photocatalysts based on zinc hydroxide and a solid solution of CdS and ZnS were prepared via the precipitation method and used for photocatalytic hydrogen production from aqueous solutions of inorganic (Na₂S/Na₂SO₃) and organic (ethanol) sacrificial agents. The photocatalysts were tested in cyclic experiments for hydrogen evolution and studied using X-ray diffraction (XRD), UV-Vis diffuse reflectance spectroscopy, high-resolution transmission electron microscopy (HRTEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) techniques. Different transformations of the β-Zn(OH)₂ co-catalyst were observed in the presence of inorganic and organic sacrificial agents; namely, ZnS was formed in Na₂S/Na₂SO₃ solution, whereas the formation of ε-Zn(OH)₂ was detected in solution with ethanol. The composite Zn(OH)₂/Cd_1−xZn_xS photocatalysts have great potential in various photocatalysis processes (e.g., hydrogen production, CO₂ reduction, and the oxidation of organic contaminants) under visible light.

The nature of the sacrificial agent affects the transformations of a Zn(OH)₂ co-catalyst during photocatalytic hydrogen production.     

Local and Remote Effects of Mesenchymal Stem Cell Administration on Skin Wound Regeneration

Wound healing is an important medical problem. We evaluated the efficacy of locally administered mesenchymal stem cells (MSCs) isolated from human umbilical cords on the dynamics of skin wound healing. The study was conducted on the backs of Wistar rats, where two square wounds were created by removing all layers of the skin. Four groups were studied in two series of experiments: (1) a Control_NaCl group (the wounds were injected with 0.9% NaCl solution) and a Control_0 group (intact wounds on the opposite side of the same rat’s back); (2) an MSC group (injected MSCs, local effect) and a Control_sc group (intact wounds on the opposite side of the back, remote MSC effect). The area and temperature of the wounds and the microcirculation of the wound edges were measured. Histological and morphometric studies were performed on days 3 and 7 after the wounds were created. The results showed that the injection trauma (Control_NaCl) slowed the regeneration process. In both MSC groups (unlike in either control group), we observed no increase in the area of the wounds; in addition, we observed inhibition of the inflammatory process and improved wound regeneration on days 1–3 in the remote group and days 1–5 in the local (injected) group. The MSC and Control_sc groups demonstrated improved microcirculation and suppression of leukocyte infiltration on day 3. On day 7, all the studied parameters of the wounds of the Control_0 group were the same as those of the wounds that received cell therapy, although in contrast to the results of the Control_ NaCl group, fibroblast proliferation was greater in the MSC and Control_sc groups. The dynamics of the size of the wounds were comparable for both local and remote application of MSCs. Thus, even a one-time application of MSCs was effective during the first 3–5 days after injury due to anti-inflammatory processes, which improved the regeneration process. Remote application of MSC, as opposed to direct injection, is advisable, especially in the case of multiple wounds, since the results were indistinguishable between the groups and injection trauma was shown to slow healing.     

Determination of sodium and potassium ions in patients with SARS-Cov-2 disease by ion-selective electrodes based on polyelectrolyte complexes as a pseudo-liquid contact phase

Nowadays, there are several methods for the detection of various bioelements during SARS-CoV-2. Many of them require special equipment, high expenses, and a long time to obtain results. In this study, we aim to use polyelectrolyte multilayers for robust carbon fiber-based potentiometric sensing to determine the ion concentration in human biofluids of COVID-19 patients. The polyethyleneimine/polystyrene sulfonate complex is hygroscopic and has the ability to retain counterions of inorganic salts. This fact makes it possible to create a flexible ionometric system with a pseudo-liquid connection. The formation of the polyethyleneimine/polystyrene sulfonate complex allows for the adhesion of a hydrophobic ion-selective membrane, and creates a Nernst response in a miniature sensor system. This approach discloses the development of miniaturized ion-selective electrodes and their future application to monitor analyte changes as micro and macroelement ions in the human body to identify correlation to SARS-CoV-2. An imbalance in the content of potassium and sodium in urine and blood is directly related to changes in the zinc content in patients with coronavirus. The proposed method for assessing the condition of patients will allow fast determination of the severity of the course of the disease.

Supramolecular assemblies based on polyelectrolyte complexes made it possible to create complex interfaces with predictable properties. Polyelectrolyte complexes serve as a pseudo-liquid contact in ion-selective electrodes.     

Improving the Conductivity of the PEDOT:PSS Layers in Photovoltaic Cells Based on Organometallic Halide Perovskites

Among conductive polymers, PEDOT films find the widest application in electronics. For photovoltaic applications, studies of their optical properties, stability, and electrical conductivity are of greatest interest. However, the PEDOT:PSS transport layers, when used in photovoltaic cells, have a high electrical resistance, which prevents solar cells from increasing their efficiency. One of the promising ways to improve their electrical properties is the use of composite materials based on them, in which the conductivity can be increased by introducing various additives. In this work, conductive polymer films PEDOT:PSS (poly (3,4-ethylenedioxythiophene):polystyrene sulfonate acid) doped with a number of amines (Pentylamine, Octylamine, Diethylamine, Aniline with carbon nanotubes) were obtained and studied. It is shown that, depending on the concentration of dopants, the electrical conductivity of PEDOT:PSS films can be significantly improved. In this case, the light transmission of the films practically does not change. The process of improving the conductivity by treating the surface of the finished film with amines, followed by heat treatment, was studied. It is assumed that the improvement in conductivity is the result of the self-assembly of monolayers of organic molecules on the surface of the PEDOT:PSS film leading to its p-doping due to intermolecular interaction.