Anterior cervical discectomy and fusion has been associated with the development of adjacent segment degeneration (ASD), with clinical incidence of approximately 3% per year. Cervical total disc arthroplasty (TDA) has been proposed as an alternative to prevent ASD.

This study assessed the applicability of loop-mediated isothermal amplification (LAMP) for the detection of leptospirosis among domesticated animals and sewage rats. Specifically, it evaluated the ability of LAMP to amplify Leptospira spp. targeting the 16s rRNA gene in boiled urine samples.

Camelina sativa (C. sativa), an oilseed species rich in poly-unsaturated fatty acids, has gained great importance as an industrial oil platform crop in recent years. Despite the potential benefits of C. sativa for bioenergy applications, limited research has been conducted to improve its agronomic qualities. Hence, a simple and efficient technique for production of transgenic C. sativa plants is warranted. In the present study, shoot apical meristems of two C. sativa cultivars (Pl650159 and Pl650161) were transformed with Agrobacterium strain 'EHA 105' harboring the enhanced green fluorescent protein (EGFP) and neomycin phosphotransferase II (nptII) genes. After two days of co-cultivation in the dark, explants were transferred to selection medium. Transgenic shoots were identified on the basis of green fluorescence and kanamycin resistance. Shoots were then rooted and transferred to potting mix soil for acclimatization. This protocol describes an efficient method to generate transgenic C. sativa plants in as little as 4 weeks.

Lentiviral-mediated gene therapy has been proposed for the treatment of a range of diseases, and due to its genome integration properties, it offers the potential for long-lasting benefit from a once-off treatment. Production methods for pre-clinical studies in animal models, and ultimately for human clinical trials, must be capable of producing large quantities of high-quality lentiviral vector in an efficient and cost-effective manner. We report here a medium-scale method (from 1.5 L to 6 L of vector supernatant) for lentiviral vector production in adherent cell cultures using the NUNC™ EasyFill™ Cell Factory™ from Thermo Fisher Scientific. Downstream purification uses a Mustang Q XT5 anion exchange capsule from Pall, and an ultracentrifugation step to concentrate the vector. This method is capable of producing lentiviral vector with concentrated titres of 108-109 TU/ml, with reduced manual handling compared to single monolayer flask methods.

The nuclear receptors are transcription factors involved in the regulation of a variety of physiological processes whose activity can be modulated by binding to relevant small molecule ligands. Their dysfunction has been shown to play a role in disease states such as diabetes, cancer, inflammatory diseases, and hormonal resistance ailments, which makes them interesting targets for drug discovery. The nuclear receptor REV-ERBα is involved in regulating the circadian rhythm and metabolism. Its natural ligand is heme and there is significant interest in identifying novel synthetic modulators to serve as tools to characterize its function and to serve as drugs in treating metabolic disorders. To do so, we established a mammalian cell-based two-hybrid assay system capable of measuring the interaction between REV-ERBα and its co-repressor, nuclear co-repressor 1. This assay was validated to industry standard criteria and was used to screen a subset of the LOPAC®1280 library and 29568 compounds from a diverse compound library. Profiling of the primary hits in a panel of counter and selectivity assays confirmed that REV-ERBα activity can be modulated pharmacologically and chemical scaffolds have been identified for optimization.

Metastatic disease is the leading cause of pancreatic ductal adenocarcinoma (PDAC) associated death. PDAC cells invade and enter the bloodstream early, before frank malignancy can be detected. Our objective was to develop an in vivo assay enabling the identification and quantification of circulating tumor cells (CTCs) from primary orthotopic PDAC tumors. Human PDAC cells expressing luciferase and green fluorescent protein were orthotopically injected into the pancreas of mice utilizing ultrasound guidance. Bioluminescent imaging was conducted to identify and track tumor growth. CTCs were then isolated and analyzed by flow cytometry to detect GFP-expressing cancer cells. Tumor growth as measured by bioluminescent imaging increased over time. The concentration of CTCs correlated with the strength of bioluminescent imaging signal. In addition, livers bearing macroscopic disease were harvested for further imaging under fluorescence stereomicroscopy and confocal microscopy, which confirmed the presence of metastases. This study represents an orthotopic animal model that reliably detects the presence of CTCs from PDAC. There is a positive correlation between the concentrations of CTCs with overall tumor burden.

Macrophage behavior is of great interest in response to tissue injury and promotion of regeneration. With increasing numbers of zebrafish reporter-based assays, new capabilities now exist to characterize macrophage migration, and their responses to biochemical cues, such as reactive oxygen species. Real time detection of macrophage behavior in response to oxidative stress using quantitative measures is currently beyond the scope of commercially available software solutions, presenting a gap in understanding macrophage behavior. To address this gap, we developed an image analysis pipeline solution to provide real time quantitative measures of cellular kinetics and reactive oxygen species content in vivo after tissue injury. This approach, termed Zirmi, differs from current software solutions that may only provide qualitative, single image analysis, or cell tracking solutions. Zirmi is equipped with user-defined algorithm parameters to customize quantitative data measures with visualization checks for an analysis pipeline of time-based changes. Moreover, this pipeline leverages open-source PhagoSight, as an automated keyhole cell tracking solution, to avoid parallel developments and build upon readily available tools. This approach demonstrated standardized space- and time-based quantitative measures of (1) fluorescent probe based oxidative stress and (2) macrophage recruitment kinetic based changes after tissue injury. Zirmi image analysis pipeline performed at execution speeds up to 10-times faster than manual image-based approaches. Automated segmentation methods were comparable to manual methods with a DICE Similarity coefficient > 0.70. Zirmi provides an open-source, quantitative, and non-generic image analysis pipeline. This strategy complements current wide-spread zebrafish strategies, for automated standardizations of analysis and data measures.

The budding yeast Saccharomyces cerevisiae is a major model system in the study of aging. Like metazoans, yeast lifespan is extended by caloric restriction and treatment with pharmacological agents which extend lifespan. A major workhorse of aging research in budding yeast is the chronological lifespan assay. Traditionally, chronological lifespan assays consist of taking regular samples of aging yeast cultures, plating out aliquots on agar, and counting the resulting colonies. This method, while highly reliable, is labor-intensive and expensive in terms of materials consumed. Here, we report a novel MTT-based method for assessing chronological lifespan in yeast. We show that this method is equal to the colony counting method in its rigorous and reliable measurement of lifespan extension in yeast as a result of caloric restriction, and is able to distinguish known long-lived and short-lived yeast strains. We have further developed this method into a high-throughput assay that allows rapid screening of potential anti-aging compounds as well as yeast strains with altered lifespan. Application of this method permits the rapid identification of anti-aging activities in yeast and may facilitate identification of materials with therapeutic potential for higher animals and, most importantly, humans.

IL23 receptor (IL23R) binding by IL23 is required for the maturation of CD4+ cells into Th17 cells and subsequent generation of IL17A and TNF. As IL23R variations contribute to AS susceptibility, we investigated the effect of IL23R variants on cytokine levels and disease measures in an AS cohort.

The aim was to evaluate the quality and acceptability of patient-reported outcome measures used to assess fatigue in patients with axial spondyloarthritis.

Emerging evidence suggests that multisensory illusions can modulate pain and can lead to changes in body perception. The aim of this study was to investigate whether contextual factors could explain the analgesic effects of multisensory body illusions on pain and body perception in people with hand OA (HOA).

Developing environment-friendly active and selective catalysts for oxidative dehydrogenation of propane for on-purpose propene synthesis is challenging despite tremendous industrial potential for this reaction. Herein, we report on catalytic activity of high surface area hexagonal boron nitride, toward oxidative dehydrogenation of propane. It shows remarkable selectivity for alkenes (∼70%) at very high conversion (of ∼50%) of propane. Propene and ethene selectivities as high as 53 and 18%, respectively, were obtained at a conversion of 52%. The catalytic activity is retained continuously for 5 h. Regeneration in ammonia brings back the catalytic activity to its full potential. Oxidation of surface B-N bonds in oxygen leads to the diminishing catalytic activity after 5 h which, on heating in ammonia, reduced back to their native form, regaining the indigenous activity. Remarkably, the addition of ammonia in the reaction feed showed stable activity for more than 100 h.

In this article, we have investigated the unusual dynamics of tert-butyl alcohol (TBA)-water and trimethylamine N-oxide (TMAO)-water binary mixtures using solvation dynamics as a tool. For this purpose, femtosecond transient absorption spectroscopy has been employed. Although these two molecules are isosteres to each other, a significant difference in water dynamics has been observed. The solvation times in TBA-water binary mixtures are found to be between 1.5 and 15.5 ps. On the contrary, we have observed very fast dynamics in TMAO-water binary mixtures (between 210 and 600 fs). Interestingly, unusual retardation in dynamics is observed at 0.10 mole fraction of TBA and TMAO in both the binary mixtures.

Design strategies for small molecular probes lay the foundation of numerous synthetic chemosensors. A water-soluble colorimetric calcium molecular probe inspired by the ionophore-based ion-selective optode is presented here with a tunable detection range (around micromolar at pH 7). The binding of Ca2+ resulted in the deprotonation of the probe and thus a significant spectral change, mimicking the ion-exchange process in ion-selective optodes. The 1:1 exchange between Ca2+ and H+ was confirmed with Job's plot. Computational studies revealed possible monomer and dimer forms of the probe-Ca2+ complexes.

Formic acid and its salts are an alternative source for hydrogen generation. In this study, we store hydrogen using the formate-bicarbonate cycle. Aqueous sodium bicarbonate is hydrogenated to form sodium formate, which can then be decomposed to release hydrogen and sodium bicarbonate. The hydrogenation step is carried out under mild conditions in the presence of a homogeneous ruthenium catalyst. Hydrogen charge is realized at 70 °C under a hydrogen pressure of 20 bar, achieving yields > 80% and turnover number > 610. The catalyst is stable and robust through numerous cycles of the hydrogenation reaction. The formate ion formed during the bicarbonate hydrogenation is assayed and quantified by ion chromatography.

"Side chain engineering" research has yielded many promising and beneficial results, with applications in various fields. However, this research did not receive sufficient focus when nature-sourced polymers are concerned. In this study, we have performed side chain engineering on chitosan, a nature-sourced polysaccharide, by coupling it with a number of aliphatic aldehydes of varying chain lengths. The side chains' length and the pursuing effect on the modified products' properties were studied in great detail. In terms of coupling yields, it was found that some substituents have displayed more favorable results than others by a factor of over 35 times. When studying the modified polymers' physical and mechanical properties, some of them were found to exhibit more rigid mechanical properties by a factor of 3.5 times than others. The effect was also expressed through self-assembly concentrations and encapsulation capabilities of the modified polymers. Remarkably, the combined experimental and calculated kinetic studies showed the results do not necessarily follow a linear progression relating to substituent chain length, but rather a parabolic pattern with a specific extremum point. This study has assisted in shedding light on the inspected phenomenon, explaining that not only steric and electronic factors but also interfacial solubility related factors govern the coupling reaction and the resulting modified polymers' properties. As chemical protocols in various academic, clinical, and industrial studies around the world slowly shift their norms toward finding safer ways for the production of novel materials and technologies, nature-sourced polymers hold great promise as virtually inexhaustible raw materials. The perfection of their chemical modification is therefore relevant now more than ever, with far-reaching and diverse applicative prospects.

A novel application of [DTBNpP] Pd(crotyl)Cl (DTBNpP = di-tert-butylneopentylphosphine) (P2), an air-stable, commercially available palladium precatalyst that allows rapid access to a monoligated state, has been identified for room-temperature, copper-free Sonogashira couplings of challenging aryl bromides and alkynes. The mild reaction conditions with TMP in dimethyl sulfoxide afford up to 97% yields, excellent functional group tolerability, and broad reaction compatibility with access to one-pot indole formation.

The valorization of olive oil mill solid wastes (OMW) has been addressed by considering it as a possible source of humic-like substances (HLSs), to be used as auxiliary substances for photo-Fenton, employing caffeine as a target pollutant to test the efficiency of this approach. The OMW-HLS isolation encompassed the OMW basic hydrolysis, followed by ultrafiltration and drying. OMW-HLS structural features have been investigated by means of laser light scattering, fluorescence, size exclusion chromatography, and thermogravimetric analysis; moreover, the capability of OMW-HLS to generate reactive species under irradiation has been investigated using spin-trap electronic paramagnetic resonance. The caffeine degradation by means of photo-Fenton process driven at pH = 5 was significantly increased by the addition of 10 mg/L of OMW-HLS. Under the mechanistic point of view, it could be hypothesized that singlet oxygen is not playing a relevant role, whereas other oxidants (mainly OH• radicals) can be considered as the key species in promoting caffeine degradation.

A new approach to develop novel hollow urchinlike copper oxide (CuO) microspheres by the hydrothermal method was reported, and zwitterionic alkyldimethylbetaine (BS) surfactants were employed as templates in the classic copper-ammonia complex systems. Effects of numerous environmental factors on the morphology of CuO particles were studied systematically, in which the concentration and structure of BS predominantly affected the developed CuO materials. It was noticed that hollow urchinlike CuO microspheres were generally formed in the presence of BS regardless of the reaction temperature and time and the source of copper ions. Generally speaking, high concentrations of BS and BS with longer chain length strongly favored the formation of hollow urchinlike CuO microspheres. The microstructures of synthesized CuO particles were studied in detail, and the corresponding formation mechanism of hollow urchinlike CuO microspheres was also proposed based on the selective adsorption of BS on the particular crystal facets of CuO crystals. Moreover, hollow urchinlike CuO microspheres showed excellent performance in the lithium-ion batteries as anode materials with a reversible capability of 511 mA h·g-1 at 0.1 C after 40 charge-discharge cycles, which was one of the best values of CuO materials reported in this field.

Realizing quantitative surface-enhanced Raman scattering (SERS) analysis is extremely helpful and challenging. Here, we utilize a facile method to synthesize spiked plasmonic nanorods with an interior gap. The Raman signal from the molecules embedded in the gap can be dramatically enhanced, leading to strong, stable, and reproducible SERS signals that can be used as an internal reference for quantitative SERS analysis. We demonstrate that the rough exterior surface has a good performance in enhancing the Raman signal of polycyclic aromatic hydrocarbon molecules adsorbed on the surface. The result shows that this method is applicable for a large range of analyte concentrations and there is an excellent linear relationship between the SERS intensity ratio and the analyte concentration (0.5-100 μM).