Proof Of Concept Research Articles

Enhancing anti-inflammatory effect of brucine nanohydrogel using rosemary oil: a promising strategy for dermal delivery in arthritic inflammation.

Brucine (BRU), an active constituent of Strychnos nux-vomica L., is one of the potential agents to control subside swelling in arthritis. However, its hydrophobic nature, poor permeation, shorter half-life, narrow therapeutic window, and higher toxicity impede its clinical applications. Hence, this investigation was aimed to develop and evaluate novel BRU loaded β-cyclodextrin (β-CD) nanosponges (BRUNs) hydrogel consisting rosemary essential oil (RO), which have been tailored for delayed release, enhanced skin permeation, and reduced irritation, while retaining anti-oxidant and anti-inflammatory activities of this bioactive. Firstly, BRUNs were fabricated by melt technique and characterized appropriately. BRUNs6 demonstrated twofold enhancement in BRU solubility (441.692 ± 38.674) with minimum particle size (322.966 ± 54.456) having good PDI (0.571 ± 0.091) and zeta potential (-14.633 ± 6.357). In vitro release results demonstrated delayed release of BRU from BRUNs6 (67 ± 4.25%) over 24h through molecular diffusion mechanism. Further, preserved anti-inflammatory (53.343 ± 0.191%) and antioxidant potential (60.269 ± 0.073%) of bioactive was observed in BRUNs6. Hence, this Ns batch was engrossed with Carbopol®934 hydrogel with RO and characterized. In vitro (release and anti-inflammatory activity), ex-vivo (skin permeability) and in vivo (carrageenan-induced inflammation) assays along with irritation study were conducted for fabricated hydrogels. Results revealed that in vitro release of BRU was further delayed from Ns hydrogel with RO (56.45 ± 3.01%) following Fickian mechanism. Considerable enhancement in skin permeability (60.221 ± 0.322µg/cm2/h) and preservation of anti-inflammatory activity (94.736 ± 2.002%) was also observed in BRUNs6 hydrogel containing RO. The irritation of BRU was found reduced (half) after its entrapped in Ns. Further, as a proof of concept, BRUNs6 hydrogel with RO effectively reduced (75.757 ± 0.944%) carrageenan-induced inflammation in rat model in comparison to pure BRU (54.914 ± 1.081%). Hence, BRUNs hydrogel with RO can be considered as a promising alternative for dermal delivery of BRU in arthritis.

Manipulating cationic ordering toward highly efficient and zero-thermal-quenching cyan photoluminescence

Developing effective design principle in crystalline materials for superior optical properties remains a critical challenge. Herein, we establish the cationic ordering strategy to build rigid framework and significantly improve phosphor performance. As a proof of concept, the NaHf2-xYx(PO4)3:Eu2+ (x = 0.05–0.2) phosphor series is constructed, which possesses absolutely cationic ordered structure and the linked outstanding photoluminescent properties. Structurally, heterovalent substitution in the lattice maintains the single-site occupancy of Na+ ions in NaHf2-xYx(PO4)3:Eu2+, as confirmed by Rietveld refinements and NMR spectroscopy. This cationic arrangement keeps the cation/vacancy ordered distribution that ensures high structural rigidity, with Debye temperature, as the indicator of rigidity, extracted from low temperature heat capacities exceeding 379 K. The resultant high structural rigidity, coupled with intrinsic defects, induces zero-thermal-quenching luminescence up to 150 °C. Moreover, the substitution of Y3+ at Hf4+ site facilitates effective Eu3+ to Eu2+ reduction, making them excellent cyan emitters with quantum efficiency exceeding 90 % and intense violet absorption. This work provides an impetus for the discovery of the currently state-of-the-art cyan phosphors for practical applications and gives insight into the structural factors influencing luminescent properties.

Phase 1 Study of CK0801 in Treatment of Bone Marrow Failure Syndromes.

An inflammatory bone marrow microenvironment contributes to acquired bone marrow failure syndromes. CK0801, an allogeneic T regulatory (Treg) cell therapy product, can potentially interrupt this continuous loop of inflammation and restore hematopoiesis. In this phase 1 dose-escalation study of CK0801 Treg cells, we enrolled patients with bone marrow failure syndromes with suboptimal response to their prior therapy to determine the safety and efficacy of this treatment for bone marrow failure syndromes. We enrolled nine patients with a median age of 57 years (range, 19 to 74) with an underlying diagnosis of aplastic anemia (n=4), myelofibrosis (n=4), or hypoplastic myelodysplasia (n=1). Patients had a median of three prior therapies for a bone marrow failure syndrome. Starting dose levels of CK0801 were 1 × 106 (n=3), 3 × 106 (n=3), and 10 × 106 (n=3) cells per kg of ideal body weight. No lymphodepletion was administered. CK0801 was administered in the outpatient setting with no infusion reactions, no grade 3 or 4 severe adverse reactions, and no dose-limiting toxicity. At 12 months, CK0801 induced objective responses in three of four patients with myelofibrosis (two had symptom response, one had anemia response, and one had stable disease) and three of four patients with aplastic anemia (three had partial response). Three of four transfusion-dependent patients at baseline achieved transfusion independence. Although the duration of observation was limited at 0.9 to 12 months, there were no observed increases in infections, no transformations to leukemia, and no deaths. In previously treated patients, CK0801 demonstrated no dose-limiting toxicity and showed evidence of efficacy, providing proof of concept for targeting inflammation as a therapy for bone marrow failure. (Funded by Cellenkos Inc.; Clinicaltrials.gov number, NCT03773393.).

Proteomics of human platelet lysates and insight from animal studies on platelet protein diffusion to hippocampus upon intranasal administration.

Human platelet lysates (HPLs) from allogeneic platelet concentrates (PCs) are biomaterials, which are rich in various trophic factors, increasingly used in regenerative medicine and biotherapy. Understanding how preparation methods influence the HPL protein profile, biological function, and clinical outcomes is crucial. Our study sheds light on the proteomes and functionality of different HPLs, with the aim of advancing their scientifically grounded clinical applications. To achieve this, PCs suspended in plasma underwent three distinct processing methods, resulting in seven HPL types. We used three characterization techniques: label-free proteomics and tandem mass tag (TMT)-based quantitative proteomics, both before and after the immunodepletion of abundant plasma proteins. Bioinformatic tools assessed the proteome, and western blotting validated our quantitative proteomics data. Subsequent pre-clinical studies with fluorescent labeling and label-free proteomics were used as a proof of concept for brain diffusion. Our findings revealed 1441 proteins detected using the label-free method, 952 proteins from the TMT experiment before and after depletion, and 1114 proteins from the subsequent TMT experiment on depleted HPLs. Most detected proteins were cytoplasmic, playing key roles in catalysis, hemostasis, and immune responses. Notably, the processing methodologies significantly influenced HPL compositions, their canonical pathways, and, consequently, their functionality. Each HPL exhibited specific abundant proteins, providing valuable insight for tailored clinical applications. Immunoblotting results for selected proteins corroborated our quantitative proteomics data. The diffusion and differential effects to the hippocampus of a neuroprotective HPL administered intranasally to mice were demonstrated. This proteomics study advances our understanding of HPLs, suggesting ways to standardize and customize their production for better clinical efficacy in regenerative medicine and biotherapy. Proteomic analyses also offered objective evidence that HPPL, upon intranasal delivery, not only effectively diffuses to the hippocampus but also alters protein expression in mice, bolstering its potential as a treatment for memory impairments.

Using models and maps to inform Target Product Profiles and Preferred Product Characteristics: the example of Wolbachia replacement

Background The global prevalence of diseases transmitted by Aedes aegypti mosquitoes, such as dengue, Zika and Yellow Fever, is increasing, but development of promising new mosquito control technologies could reverse this trend. Target Product Profiles (TPPs) and Preferred Product Characteristics (PPCs) documents issued by the World Health Organization can guide the research and development pathways of new products and product combinations transitioning from proof of concept to operational use. Methods We used high resolution global maps of the case and economic burden of dengue to derive programmatic cost targets to support a TPP for Wolbachia replacement. A compartmental entomological model was used to explore how release size, spacing and timing affect replacement speed and acceptability. To support a PPC for a hybrid suppress-then-replace approach we tested whether Wolbachia replacement could be achieved faster, more acceptably or at a lower cost if preceded by a mosquito suppression programme. Results We show how models can reveal trade-offs, identify quantitative thresholds and prioritise areas and intervention strategies for further development. We estimate that for Wolbachia replacement to be deployable in enough areas to make major contributions to reducing global dengue burden by 25% (in line with 2030 WHO targets), it must have the potential for cost to be reduced to between $7.63 and $0.24 (USD) per person protected or less. Suppression can reduce the number of Wolbachia mosquitoes necessary to achieve replacement fixation by up to 80%. A hybrid approach can also achieve fixation faster and potentially improve acceptability, but may not justify their cost if they require major new investments in suppression technologies. Conclusions Here we demonstrate the value dedicated modelling can provide for interdisciplinary groups of experts when developing TPPs and PPCs. These models could be used by product developers to prioritise and shape development decisions for new Wolbachia replacement products.

Multifunctional Meta-Devices for Full-Polarization Rotation and Focusing in the Near-Infrared

The creation of multi-channel focused beams with arbitrary polarization states and their corresponding optical torques finds effective applications in the field of optical manipulation at the micro-nanoscale. The existing metasurface-based technologies for polarization rotation have made some progress, but they have been limited to single functions and have not yet achieved the generation of full polarization. In this work, we propose a multi-channel and spatial-multiplexing interference strategy for the generation of multi-channel focusing beams with arbitrary polarization rotation based on all-dielectric birefringent metasurfaces via simultaneously regulating the propagation phase and the geometric phase and independently controlling the wavefronts at different circular polarizations. For the proof of concept, we demonstrate highly efficient multi-channel polarization rotation meta-devices. The meta-devices demonstrate ultra-high polarization extinction ratios and high focusing efficiencies at each polarization channel. Our work provides a compact and versatile wavefront-shaping methodology for full-polarization control, paving a new path for planar multifunctional meta-optical devices in optical manipulation at micro–nano dimensions.

Proof of concept of an experimental prototype for the prevention of parastomal hernia.

The aim of this study was to analyse the complications and problems associated with the use of an experimental prototype designed for the prevention of parastomal hernia (PSH), one of the most frequent complications in ostomates. A single-centre, non-comparative, proof-of-concept interventional pilot study of an experimental prototype designed to be used in conjunction with an abdominal compression binder to prevent PSH was conducted. The "Ostomy Fixation Device for Hernia Prevention" (patent P201531826) is a semi-rigid ostomy protector, to be used in conjunction with a compression binder. It is designed to adapt to the dimensions of standard ostomy bags from different brands and serves to transmit, in a localised manner, the support coming from the compression binder in the peristomal area without putting pressure on the collection bag. The main outcome measures were efficacy, safety, and patient-users' opinion/perception. Ten patients were studied for 12 months. Mean age was 61 years (± 11.59), 70% (7) were male, 80% (8) ostomised for colorectal cancer, 90% (9) underwent planned surgery and 80% (8) had a colostomy. the incidence of HPE was 10% (1). no participant experienced pain, discomfort, itching, stinging, leakage, pouch detachment, allergy to components, or injury to the stoma or peristomal skin due to rubbing or pressure. 90% (n = 9) were considered "very satisfied" or "satisfied" with the device. An innovative device designed in collaboration between healthcare professionals and end-users has been shown to be safe and effective in reducing PSH in the group of ostomates studied.

Antimicrobial Susceptibility Testing for Colistin: Extended Application of Novel Quantitative and Morphologic Assay Using Scanning Electron Microscopy

Background. Colistin (Polymyxin E) has reemerged in the treatment of MDR Gram-negative infections. Traditional Colistin AST methods have long turnaround times and are cumbersome for routine use. We present a SEM-AST technique enabling rapid detection of Colistin resistance through direct observation of morphological and quantitative changes in bacteria exposed to Colistin. Methods. Forty-four Gram-negative reference organisms were chosen based on their Colistin susceptibility profiles. Bacterial suspensions of ∼107 CFU/mL were exposed to Colistin at EUCAST-ECOFF, with controls not exposed, incubated at 37°C, and then sampled at 0, 15, 30, 60, and 120 minutes. Phosphotungstic Acid (PTA) staining was applied, followed by SEM imaging using Hitachi TM4000PlusII-Tabletop-SEM at ×2000, ×5000 and ×7000 magnifications. Bacterial viability analysis was performed for all conditions by quantifying viable and dead organisms based on PTA-staining and morphologic changes. Results. We identified a significant drop in the percentage of viable organisms starting 30 minutes after exposure in susceptible strains, as compared to nonsignificant changes in resistant strains across all tested organisms. The killing effect of Colistin was best observed after 120 minutes of incubation with the antibiotic, with significant changes in morphologic features, including bacterial inflation, fusion, and lysis, observed as early as 30 minutes. Our observation matched the results of the gold standard-based broth microdilution method. Conclusions. We provide an extended application of the proof of concept for the utilization of the SEM-AST assay for Colistin for a number of clinically relevant bacterial species, providing a rapid and reliable susceptibility profile for a critical antibiotic.

P.083 Use of jugular venous pressure to optimize outcomes of vestibular schwannoma resection: a review of the literature and proof of concept

Background: Surgical resection of vestibular schwannoma (VS) is often curative if gross total resection is achieved, however, it is a delicate procedure with high risk to the facial nerve. With retrosigmoid approach for resection, the head is positioned to maximize lateral head rotation and neck flexion in order to optimize the surgical field. However, this may inadvertently occlude cerebral venous drainage, elevating intracranial pressure (ICP) and increasing intraoperative bleeding. Methods: Here, we review relevant literature regarding the effects of head rotation and neck flexion on internal jugular vein (IJV) occlusion and ICP, and highlight the notion that head rotation and flexion may occlude the ipsilateral IJV, increasing ICP. Subsequently, we propose a novel technique using continuous, real-time monitoring of jugular bulb pressure (JBP) to detect obstructions in jugular venous flow and guide optimal head positioning prior to VS resection. Results: As proof of concept, we present a case in which JBP monitoring was employed to optimize head positioning prior to a VS resection, which shows a significant reduction in JBP compared to traditional positioning. Conclusions: This innovative approach offers promise in enhancing the safety and efficacy of intracranial surgery for VS and potentially other neurosurgical procedures.

Programmable metasurfaces with high angular stability for arbitrarily-polarized obliquely-incident waves

Abstract Programmable metasurfaces show enormous potential in real-time electromagnetic (EM) manipulation and their stable responses for variable EM waves including incident angle and polarization changes is crucial for related applications. However, the demonstrated programmable metasurfaces are commonly considered in normal-incident wave with fixed polarization; how to achieve stable performance in more realistic scenarios of wide-angle and full-polarized wave incidences is still a challenge. Here, we propose and realize a wide-angle and full-polarization programmable metasurface, which can generate stabilized amplitude and phase responses at both transverse electric (TE) and transverse magnetic (TM) oblique incidences. These are achieved by introducing the metallic walls around the metasurface element to reduce element coupling, which greatly improves the angular stability of its reflection responses. The mechanisms of angular insensitivity are analyzed comprehensively, and as a proof of concept, the obliquely-incident beam steering, large-angle beam scanning and oblique vortex beam emitting are demonstrated on this programmable metasurface. Both the simulation and experimental results demonstrate that the programmable metasurface can operate well at both TE and TM wide-angle oblique incidences in the upper half space. Our work offers an actual route for improving the angular stability and polarization insensitivity of metasurface, which could push it one step closer towards more complicated applications.

Nanoparticle ZnS:Ag/6LiF - A new high count rate neutron scintillator with pulse shape discrimination

Abstract A neutron sensitive scintillator employing nanoparticles of ZnS:Ag has been developed for the first time. Pulse shape differences between neutron and gamma signals were observed in this material and neutron-gamma discrimination was applied. With initial signal processing parameters, gamma sensitivities of 8.5 × 10-6 to 60Co gammas were achieved. The average primary decay of neutron scintillation in nanoparticle ZnS:Ag/6LiF was measured to be 18ns, and afterglow was significantly suppressed in comparison to standard ZnS:Ag/6LiF scintillators that employ micron sized ZnS:Ag. Fast decay times and minimal afterglow indicate potential for use in high count rate capability applications. Prospective count rate capabilities were investigated here as proof of concept, with rates of 1.12Mcps measured for a single readout channel with less than 3.5% count loss. This is approximately 70 times greater than the count rate capability of the current standard ZnS:Ag/6LiF scintillation detectors. With improvements to signal processing and scintillator composition, nanoparticle ZnS:A/6LiF could be a promising candidate for future high rate capability neutron detectors.

Poly(aniline‐co‐pyrrole) as Adsorbent for Removal of Pharmaceuticals: Preparation, Characterization, Kinetics, Isotherms and Application on Sample Preparation

AbstractHerein, poly(aniline‐co‐pyrrole) has been synthetized and investigated to adsorb phenylbutazone (PBZ), ceftiofur (CEF) and prednisolone (PRD) in aqueous medium as proof of concept. After proper adsorbent characterization, some studies were carried out in batch process, i. e., contact time, pH, and concentration. The adsorption mechanism has been conducted using kinetic and isothermal models. It was found that the reaction equilibrium was reached in 20 min for all analytes. The best fit for the experimental data were obtained employing the models of dual‐site Langmuir–Freundlich, Fritz–Schlunder and Langmuir, with R2=0.999, 0.952 and 0.974, for PBZ, CEF and PRD, respectively. For PBZ, CEF and PRD, poly(aniline‐co‐pyrrole) provided a Qmax=238.8 mg g−1 and Kmax=0.086 L g−1, Q=0.028 mg g−1 and Kmax=3.258 L g−1, Q=13.66 mg g−1 and KL=0.005 L g−1, respectively. Finally, the copolymer was evaluated in solid phase extraction, which showed recovery around 70 % for PBZ, offering also exciting opportunities for further application in the separation process.

Rotating twisted templates for imprinting polarization gratings with a sub- to dozen-micron period

In this Letter, we report and experimentally demonstrate what is to our knowledge a novel scheme for imprinting polarization gratings (PGs) with a pair of templates. Compared with the traditional method that a single template can only imprint PG with a single period, cascading two templates can control the period of imprinted PG at will. However, the low diffraction efficiency is inevitably caused by cascading two templates. Therefore, a rigorous coupled wave analysis (RCWA) is adopted to design a multi-twisted template to address this challenge. As a proof of concept, two multi-twisted templates with a period of 1.6 μm were fabricated, and PGs with a large period range from 0.4 to 48.6 μm were successfully imprinted. The proposed scheme is expected to enable rapid, robust, and high-quality mass production of beam steering, large-angle deflectors, and diffractive optical couplers.

E.2 Proof of concept for liquid biopsy: positive correlation between extracellular vesicles shed by high grade gliomas and volume of hypervascular tumour tissue on MRI

Background: High grade gliomas (HGGs) shed extracellular vesicles (EVs) into the bloodstream. EV-derived RNA (EV-RNA) can be detected in plasma, making it a potential biomarker for HGG recurrence after treatment. We sought to establish a baseline relationship between EV-RNA in plasma and hypervascular HGG tissue on MRI. Methods: Eight patients with a new diagnosis of HGG had measurements of plasma EV-RNA and contemporaneous dynamic susceptibility contrast (DSC) MRI. Patient-specific median signal intensity of corpus callosum (mSI-CC) was determined from 10 measurements on the relative cerebral blood volume (rCBV) map. Tumour tissue with signal intensity > mSI-CC and > 2x, > 3x, > 4x and > 5x mSI-CC was segmented on the rCBV map. EV-RNA plasma concentration was correlated with tissue volumes. Results: Pearson correlation showed a significant positive relationship between EV-RNA plasma concentration and tissue volume with signal intensity > mSI-CC (r(6) = 0.899, p = 0.002). No significant relationship could be detected for progressively smaller tissue volumes with signal intensity > 2x, > 3x, > 4x and > 5x mSI-CC. Conclusions: EV-RNA plasma concentration correlates strongly with the total volume of hypervascular HGG tissue on DSC MRI at baseline and merits further evaluation as a biomarker of tumour behaviour in longitudinal imaging studies.

Photodissociation and induced chemical asymmetries on ultra-hot gas giants

Context. Recent observations have resulted in the detection of chemical gradients on ultra-hot gas giants. Notwithstanding their high temperature, chemical reactions in ultra-hot atmospheres may occur in disequilibrium, due to vigorous day-night circulation and intense UV radiation from their stellar hosts. Aims. The goal of this work is to explore whether photochemistry is affecting the composition of ultra-hot giant planets, and if it can introduce horizontal chemical gradients. In particular, we focus on hydrogen cyanide (HCN) on WASP-76 b, as it is a photochemically active molecule with a reported detection on only one side of this planet. Methods. We used a pseudo-2D chemical kinetics code to model the chemical composition of WASP-76 b along its equator. Our approach improved on chemical equilibrium models by computing vertical mixing, horizontal advection, and photochemistry. Results. We find that the production of HCN is initiated through the thermal and photochemical dissociation of CO and N2 on the day side of WASP-76 b. The resulting radicals are subsequently transported to the night side via the equatorial jet stream, where they recombine into different molecules. This process results in an HCN gradient with a maximal abundance on the planet’s morning limb. We verified that photochemical dissociation is a necessary condition for this mechanism, as thermal dissociation alone proves insufficient. Other species produced via night-side disequilibrium chemistry are SO2 and S2. Conclusions. Our model acts as a proof of concept for chemical gradients on ultra-hot exoplanets. We demonstrate that even ultra-hot planets can exhibit disequilibrium chemistry and recommend that future studies do not neglect photochemistry in their analyses of ultra-hot planets.

Process and Impact of Development of an Adolescent Emergency Psychiatry Unit at a Large Urban Hospital.

Boarding of adolescent patients with mental health concerns requiring ongoing observation and treatment is of increasing concern across US emergency departments. The objective was a proof of concept of developing an adolescent psychiatric emergency unit and assessment of the impact of this unit on lengths of stay (LOS). We describe the creation of the unit designed to allow safe assessment and boarding of patients, and appropriate interventions and services, while arranging transfer to inpatient facility or safe discharge home. Using a precreation and postcreation analysis and comparison with a similar facility that did not create such a unit, we utilized linear regression to investigate the primary outcome of total length of stay and secondary outcomes of psychiatric emergency department and pediatric emergency department length of stay for both unit-eligible patients and all patients. The overall length of stay was not associated with a statistically significant change for unit-eligible patients; however, there was a significant decrease in the pediatric emergency department LOS for unit-eligible patients. This was associated with a decrease in beds lost to boarding in the pediatric emergency department of 544 hours per month. Creation of an adolescent psychiatric emergency unit without allotment of significant additional resources is an option to decrease pediatric emergency department boarding times for adolescent patients requiring ongoing emergent therapy for mental health concerns.

Microfabrication through Self‐Ordering of Cracks: Mechanism, Upscaling and Application for Transparent Electrodes

AbstractWhen drying a colloidal solution, cracks appear in the resulting colloidal film. In certain cases, spontaneous order is observed, and cracks form arrays of periodic patterns. Although this phenomenon might be envisioned as a patterning method, overcoming practical challenges is necessary to transform it into a technological tool for microfabrication. This study explores various technological aspects aimed at leveraging the self‐assembly of cracks as a scalable microfabrication tool for large‐scale device production. Through a series of analyses, including time‐resolved Grazing‐Incidence Small‐Angle X‐Ray Scattering (GISAXS), it is offered novel insights into controlling the crack self‐ordering mechanism, minimizing defects, and implementing strategies for large‐scale patterning and pattern transfer. The process proves to be surprisingly robust, maintaining its efficacy with the same colloidal solution even after two years. By introducing biphasic dip‐coating, large‐scale crack patterns up to 100 cm2, while preserving their periodicity and ordering is achieved. As a proof of concept, the use of crack‐patterned colloidal films as masks for fabricating metallic sub‐micrometer objects, that serve as transparent electrodes with adjustable transparency and conductivity is showcased. Overall, this method presents significant advantages over conventional lithography, being cost‐effective, versatile, environmentally friendly, and scalable, thereby offering new perspectives for diverse applications requiring cost‐effective and large‐scale patterning.

#1805 Renal progenitor cells isolated from urine of IgA nephropathy patients formed renal spheroids that can recapitulate the typical IgA1 deposition

Abstract Background and Aims Recent developments in stem cell biology have made it possible to create spheroids and kidney organoids from human and mouse pluripotent stem cells and renal stem cells. Spheroids are a suitable in vitro model for studies of drug studies and can transform into organoids whose rapidly developing field has proven useful in simulating nephrotoxicity, kidney disorders, and kidney development. However, to date in all studies the generation of organoids and tubuloids required the use of sophisticated and complex differentiation protocols involving external administration of several growth factors and needs compliance with a precise and specific timescale. Here we showed for the first time that human adult renal progenitor cells (ARPCs) can be isolated from urines of healthy subjects (HS) or IgA Nephropathy (IgAN) patients and can form spheroids generating very long tubule-like structures naturally, without the external use of chemokines or growth factors to artificially induce the process and recapitulating the IgA1 deposition that is the typical characteristic of the disease. Method Spheroids and tubule-like structures were characterized by whole-mounting and frozen section immunofluorescence (IF) assays and by Flow Cytometry Analysis using CD133, NanoG, Oct3/4, GATA-3, SSEA4, CD249, Aminopepidase N, ZO-1, Uromodulin, and Lotus Tetragonolobus Lectin (LTL) antibodies. ELISA was used to determine the renin secretion and CAM assays to determine ARPCs and ARPC-derived Spheroids angiogenic properties. PKH-26 labeling was used for their in vitro and in vivo tracking. Serum of HS and IgAN patients in culture with spheroids was used to recapitulate the disease and IgA1 deposits were measured by IF. Results ARPC-derived spheroids reflected the renal progenitor properties and recapitulated organoids generating very long tubule-like structures that expressed structural and functional renal tubule markers such as CD249, Aminopepidase N, ZO-1, Uromodulin, and LTL and in some cases shared many structural similarities with some regions of nephrons, such as the distal convoluted tubule, the loop of Henle and proximal convoluted tubules. Moreover, ARPC-derived spheroids secreted elevated levels of renin and, as the cellular counterpart, showed angiogenic properties. To study the proof of concept that ARPC-derived spheroids can be used to establish in vitro models recapitulating renal diseases, we generated IgAN patient-specific renal spheroids using the ARPC mixed cell population method. ARPC-derived spheroids of IgAN patients showed deposits of IgA1 already after four days of culture with patient serum (p = 0.004) and, with a different pattern, after 8 and 15 days (p = 0.0029 and p = 0.0012, respectively). The IgA1 deposition resembled that in the glomeruli of patients diagnosticated with IgAN. In contrast, no positive signal was detected in either IgAN spheroids in culture with FBS, or with inactivated IgAN serum, as well as in spheroids derived from urine of HS in culture with IgAN serum. Conclusion The ability of ARPCs to form spheroids and differentiate into tubular-like structures without the need for external chemokines or growth factors is a significant advancement in the field, even more considering that these cells can be isolated from the urine of patients. These findings open up new possibilities for the regenerative medicine in studying kidney development, disease mechanisms, and regenerative therapy development for renal disorders.

Textural Precursor Compositions Harvested for Independent Signal Generators: Scaling Micron-Sized Flower-Like Metal-Organic Frameworks as Amplifying Units for Dual-Mode Glycoprotein Assay.

In this work, a micron-sized flower-like metal-organic frameworks (MOFs)-based boronate-affinity sandwich-type immunoassay was fabricated for the dual-mode glycoprotein assay. For proof of concept, the flower-like MOFs were synthesized from transition Cu nodes and tetrakis (4-carboxyphenyl) porphyrin (TCPP) ligands by spontaneous standing assembly. In addition, the specificity toward glycoprotein involved the antigen recognition as well as covalent bonding via the boronate-glycan affinity, and the immediate signal responses were initiated by textural decomposition of the flower-like MOFs. Intriguingly, Cu nodes, of which the valence state is dominant by CuI species, can endow the Fenton-like catalytic reaction of the fluorogenic substrate for generating fluorescence signals. For benefits, TCPP ligands, in which each TCPP molecule has four guest donors, can provide multiple valences for the assembly of cyclodextrin-capped gold nanoparticles via host-guest interaction for colorimetry output. Albeit important, the scaling micrometer patterns for the flower-like MOFs carrying numerous Cu nodes and TCPP ligands can also function as amplifying units, signifying the output signal. The detection limit of the dual-mode glycoprotein assay can reach 10.5 nM for the fluorescence mode and 18.7 nM for the colorimetry mode, respectively. Furthermore, the merits of harvesting different signal generators toward the multimodal readout patterns can allow the mutual verification and make the analytical results more reliable. Collectively, our proposed assay may offer a new idea in combining the inherent textural merits from MOFs for dual signal generators, which can also emphasize accurate detection capability for glycoprotein assay.

A double-blinded randomised controlled study of fluid restriction versus liberal fluid during induction of labour: A pilot study.

Excess intravenous fluid for women requiring an induction of labour may adversely affect the duration of labour and maternal/neonatal outcomes. This study aimed to determine the difference in duration of labour and outcomes with a low background infusion rate, compared to liberal background intravenous fluid management. A double blind randomised controlled pilot study was performed on 200 women who underwent induction of labour at a single institution. Women were randomised to an intravenous rate of 40 mL/h versus 250 mL/h of Hartmann's solution. Fluid boluses were strictly controlled to limit bias. This trial was registered with the Australian clinical trial registry: ACTRN12621001298808. Analysis of the total amount of fluid received showed good separation with Group 1 (40 mL/h) receiving 1,736 mL less than Group 2 (250 mL/h), median (interquartile range) 841 mL (458, 1691) versus 2,577 mL (1620, 4326) (P < 0.001). Median duration of labour was shorter in Group 1 by 24 min (P = ns). Subset analysis of nulliparous women showed that duration of labour was shorter in Group 1 by 83.5 min (P = ns). As this was a pilot study, a significant difference in duration of labour or secondary outcomes was not seen. Given the increasing numbers of nulliparous women having an induction of labour, potential for adverse maternal and neonatal outcomes and the associated higher rate of operative birth, this study guides power calculations and supports proof of concept for future research into optimum fluid management during induction of labour for these women.