Single-Cell Microgels for Diagnostics and Therapeutics

Cell encapsulation within hydrogel droplets is transforming what is feasible in multiple fields of biomedical science such as tissue engineering and regenerative medicine, in vitro modeling, and cell-based therapies. Recent advances have allowed researchers to miniaturize material encapsulation complexes down to single-cell scales, where each complex, termed a single-cell microgel, contains only one cell surrounded by a hydrogel matrix while remaining <100 μm in size. With this achievement, studies requiring single-cell resolution are now possible, similar to those done using liquid droplet encapsulation. Of particular note, applications involving long-term in vitro cultures, modular bioinks, high-throughput screenings, and formation of 3D cellular microenvironments can be tuned independently to suit the needs of individual cells and experimental goals. In this progress report, an overview of established materials and techniques used to fabricate single-cell microgels, as well as insight into potential alternatives is provided. This focused review is concluded by discussing applications that have already benefited from single-cell microgel technologies, as well as prospective applications on the cusp of achieving important new capabilities.     

Resveratrol as Chemosensitizer Agent: State of Art and Future Perspectives

Resistance to chemotherapy still remains a major challenge in the clinic, impairing the quality of life and survival rate of patients. The identification of unconventional chemosensitizing agents is therefore an interesting aspect of cancer research. Resveratrol has emerged in the last decades as a fascinating molecule, able to modulate several cancer-related molecular mechanisms, suggesting a possible application as an adjuvant in cancer management. This review goes deep into the existing literature concerning the possible chemosensitizing effect of resveratrol associated with the most conventional chemotherapeutic drugs. Despite the promising effects observed in different cancer types in in vitro studies, the clinical translation still presents strong limitations due to the low bioavailability of resveratrol. Recently, efforts have been moved in the field of drug delivery to identifying possible strategies/formulations useful for a more effective administration. Despite the necessity of a huge implementation in this research area, resveratrol appears as a promising molecule able to sensitize resistant tumors to drugs, suggesting its potential use in therapy-refractory cancer patients.     

Antibacterial and photocatalytic behaviour of green synthesis of Zn0.95Ag0.05O nanoparticles using herbal medicine extract

The present work aimed to synthesize Zn_0.95Ag_0.05O (ZnAgO) nanoparticles using rosemary leaf extracts as a green chemistry method. The characterization of Ag-doped ZnO nanoparticles was performed by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), and ultraviolet–visible spectrophotometry (UV–visible). The XRD, FTIR, and UV–visible spectra confirmed the formation of the presence of hexagonal ZnAgO nanoparticles. FESEM micrograph shows that the nanoparticles have been distributed homogeneously and uniformly. The morphology of ZnAgO nanoparticles is quasi-spherical configuration. Also, the mean particle size is in the range of 22–40 nm. The photocatalytic degradation of methylene blue in the presence of Ag-doped ZnO nanoparticles is nearly 98.5% after exposing 100 min. The ultraviolet lamp was used as the light source for photocatalyst degradation. The disc diffusion method was chosen to study the antibacterial activity of as-synthesized ZnAgO nanoparticles. Antibacterial activity of Zn_0.95Ag_0.05O nanoparticles against Staphylococcus aureus and Escherichia coli revealed that the as-synthesized ZnAgO nanoparticles were efficient in inhibition of bacterial growth.     

Effects of ultrafiltration combined with high-pressure processing,ultrasound and heat treatments on the quality of a blueberry–grape–pineapple–cantaloupe juice blend

This study investigated the effect of ultrafiltration (UF) combined with high-pressure processing (HPP) at 550 MPa, 25 °C for 5 min, ultrasound (US) at 520 W, 40 °C for 10 min and heat treatment (HT) at 90 °C for 3 min on the microbial, physicochemical and sensory properties of a blueberry–grape–pineapple–cantaloupe juice blend during 104 days of storage at 4 °C. After UF, the shelf life of the HPP- and US-treated clear juice blends were 104 and 72 days during the storage at 4 °C respectively. HPP, US and HT treatment minimally affected the anthocyanin and total phenol contents, while HPP better maintained the ascorbic acid levels and sensory properties in the clear juice blend during the storage. Therefore, HPP combined with UF was identified as a prospective processing technique in the fruit juice industry.     

Reproductive tract disease in Irish grazing dairy cows: Retrospective observational study examining its association with reproductive performance and accuracy of 2 diagnostic tests

The detection of reproductive tract disease (RTD) 3 wk postpartum is important because of its effect on subsequent reproductive outcomes. Numerous methods for the diagnosis of RTD are described, some of which are more practical and instantaneous in terms of diagnosis. Two of these methods involve identification of purulent vaginal discharge (PVD) and evidence of ultrasonographic uterine changes indicative of endometritis (UE). The objectives of our retrospective observational study were (1) to assess the association of PVD or UE score at the prebreeding examination (PBE) with the hazard of pregnancy within the subsequent breeding season; (2) to determine the test sensitivity (Se) and specificity (Sp) at the point of sampling of both tests using a Bayesian latent class model; and (3) to determine the effect of varying positivity thresholds on test accuracy. To achieve these objectives, we analyzed an initial data set of 5,049 PBE from 2,460 spring-calved cows in 8 herds between 2014 and 2018. Each PBE was conducted once between 25 and 86 d in milk. At each PBE, vaginal discharge was obtained with a Metricheck device (Simcro) whereas uterine contents were assessed using transrectal ultrasonography. Purulent vaginal discharge was scored on a scale of 0 to 3 depending on discharge character, and UE was scored on a scale of 0 to 4 depending on the presence and consistency of intraluminal fluid. Cows with scores of ≥2 in either test had received treatment. Fertility data were available from 4,756 PBE after data exclusion. The association between PVD or UE score at the PBE and subsequent hazard of pregnancy was analyzed using a Cox proportional hazards model. Cows with a PVD score of 2 or 3 were less likely to conceive than cows with a PVD score 0 [score 2 hazard ratio (HR) = 0.74; 95% confidence interval (CI): 0.59–0.94; score 3 HR = 0.65; 95% CI: 0.51–0.84]. Cows with a UE score of 1, 2, 3, or 4 were less likely to conceive than cows with a UE score of 0 (score 1 HR = 0.82; 95% CI: 0.73–0.93; score 2 HR = 0.79; 95% CI: 0.62–1.00; score 3 HR = 0.43; 95% CI: 0.43–0.90; score 4 HR = 0.39; 95% CI: 0.26–0.58). To determine the Se and Sp of PVD or UE score for diagnosis of RTD at the time of PBE, a Bayesian latent class model was fitted on 2,460 individual cow PBE. Flat priors were used for the Se and Sp of UE, whereas informative priors were used for PVD Se (mode = 65%, 5th percentile = 45%) and Sp (mode = 90%, 5th percentile = 80%) and RTD prevalence (mode = 20%, 5th percentile = 10%). Posterior estimates (median and 95% Bayesian probability intervals; BPI) were obtained using ‘rjags' (R Studio). The optimal test thresholds (PVD and UE score ≥1) were selected by assessing the effect of different thresholds on test estimates and using a misclassification cost analysis. Based on these, median (95% BPI) Se for PVD and UE score ≥1 were 44% (29–60%) and 67% (33–100%), respectively. Median Sp for PVD and UE score ≥1 were 90% (86–93%) and 91% (86–93%), respectively. Higher scores in both tests were associated with impaired fertility, and UE scoring with a threshold of ≥1 had the highest test Se and Sp estimates although test Se was conditional on days in milk when the PBE occurred.     

Scallops as a new source of food protein: high-intensity ultrasonication improved stability of oil-in-water emulsion stabilised by myofibrillar protein

In this study, the effect of high-intensity ultrasound (HIUS) (200 and 400 W for 0, 5, 10 and 15 min respectively) on conformational changes, physicochemical, rheological and emulsifying properties of scallop (Patinopecten yessoensis) myofibrillar protein (SMP) was investigated. HIUS-treated SMP had lower α-helix content and higher β-sheet content compared with the native SMP. HIUS treatment induced the unfolding of SMP and increased the surface hydrophobicity. The particle size of SMP decreased and the absolute zeta-potential increased after ultrasonication, which in turn increased the solubility of SMP. The conformational changes and the improvement of physicochemical properties of SMP increased the ability for SMP to lower the interfacial tension at the oil–water interface and increased the percentage of adsorbed protein. As a result, the emulsifying properties, rheological properties of SMP and storage stability of emulsions were also improved. In conclusion, HIUS treatment has future potential for improving the emulsifying properties of SMP.     

Synergistic effect of high-intensity ultrasound and β-cyclodextrin treatments on browning control in apple juice

This work evaluated the synergistic effects of combined high-intensity ultrasound (HIU) with β-cyclodextrin (β-CD) treatments on inhibiting browning of apple juice and explored the mechanism through simulation system. The combined treatment of 300 W HIU with 0.006 g mL⁻¹ β-CD had a synergistic impact on maintaining juice colour, resulting in a 39.06% reduction in browning degree, only a 36.64% decrease in total phenolic content, and a 17.82% reduction in PPO activity. The inhibition of enzymatic browning in simulated system revealed that HIU suppressed the enzyme (Polyphenol oxidase, PPO) and β-CD inhibited enzyme (PPO) and embedded substrate (polyphenol). The results of spectroscopic analysis showed that the particle-size distribution of PPO narrowed, the content of α-helix in the secondary structure increased, the fluorescence intensity increased, and the maximum wavelength was red-shifted after HIU and β-CD treatment. Changes in structure could further result in PPO activity loss. Hence, the combined treatment could synthetically alleviate the browning of apple juice.     

Characterization of Dog Glutathione Transferase P1-1, an Enzyme Relevant to Veterinary Medicine

Glutathione transferases (GSTs) form a family of detoxication enzymes instrumental in the inactivation and elimination of electrophilic mutagenic and carcinogenic compounds. The Pi class GST P1-1 is present in most tissues and is commonly overexpressed in neoplastic cells. GST P1-1 in the dog, Canis lupus familiaris, has merits as a marker for tumors and as a target for enzyme-activated prodrugs. We produced the canine enzyme CluGST P1-1 by heterologous bacterial expression and verified its cross-reactivity with antihuman-GST P1-1 antibodies. The catalytic activity with alternative substrates of biological significance was determined, and the most active substrate found was benzyl isothiocyanate. Among established GST inhibitors, Cibacron Blue showed positive cooperativity with an IC₅₀ value of 43 nM. Dog GST P1-1 catalyzes activation of the prodrug Telcyta, but the activity is significantly lower than that of the human homolog.     

Effects of ultrasound-assisted resting on the qualities of whole wheat dough sheets and noodles

The effects of resting under ultrasonic treatment on the properties of whole wheat dough sheets and noodles were investigated. Compared with the control group, the resting time to reach the maximum breaking force and extensibility of whole wheat dough sheets treated with ultrasound was shortened by 20 min. The proportion of strongly bound water (T₂₁) decreased, while the proportion of weakly bound water (T₂₂) increased during resting, and the ultrasonic treatment could accelerate this moisture redistribution in the dough. The extent of the increase in gluten macropolymer and the decrease in –SH content of ultrasound-treated wheat dough was higher than that of the control one during the initial 5–15 min resting, which could be related to the improvement of whole wheat dough extensibility. Whole wheat noodles showed a better breaking distance for 10 min of ultrasonic-assisted resting. Ultrasonic treatment could be used to reduce the resting time of whole wheat dough sheets and improve the texture properties of noodles.     

Effects of high-intensity ultrasound treatment on physiochemical properties of caseins-cyanidin-3-galactoside conjugates

The application of protein-polyphenol complexes is increasing and broadening in recent years. Due to their unique properties, these complexes are attracting increased research interest. In this study, the cyanidin-3-galactoside was grafted to the caseins using free radicles induced by ultrasound. The formation of conjugation between caseins and cyanidin-3-galactoside was confirmed by the decrease in free amino groups and sulfhydryl content. Conjugation with cyanidin-3-galactoside resulted in aggregation of caseins, which also led to increase in particle size, relative fluorescence intensity and random coil. Conjugation also disrupted the hydrogen bonds and decreased the electrostatic repulsion and hydrophobic interactions between caseins. The caseins-cyanidin-3-galactoside conjugate had better emulsifying, gelation properties and lower thermal stability than that of caseins.