Upstream Process Protocol for MSCs Isolated from Different Human-Based Tissue Origins.

This chapter introduces the increasing significance of mesenchymal stromal/stem cell (MSC) production in regenerative medicine and cellular therapeutics, outlines the growing interest in MSCs for various medical applications, and highlights their potential in advanced therapy medicinal products (ATMPs) and the advancements in cell culture technologies that have facilitated large-scale MSC production under Good Manufacturing Practices (GMP), ensuring safety and efficacy. This chapter describes an optimized upstream protocol for laboratory-scale MSC production from different tissue sources. This protocol, conducted in flasks, controls critical parameters and lays the foundation for downstream processing to generate ATMPs. This comprehensive approach underscores the potential of MSCs in clinical applications and the importance of tailored production processes.

Coffee-Derived Exosome-Like Nanoparticles: Are They the Secret Heroes?

BACKGROUND: Regular coffee consumption has beneficial and preventative effects on liver and chronic neurodegenerative diseases. However, the studies performed with the ingredients found in coffee beverages have not clarified the responsible mechanisms. Exosomes are small, membrane-coated cargo packages secreted by prokaryote and eukaryote cells. Exosomes regulate intercellular communication and affect cellular metabolic activities even among different species. In this study, we aimed to isolate and characterize the edible plant-derived exosome-like nanoparticles from roasted hot coffee beverages, hypothesizing that the edible plant-derived exosome-like nanoparticles were responsible for the beneficial effects of coffee. METHODS: Size exclusion chromatography and commercial kits were used for the isolation process. Efficient coffee edible plant-derived exosome-like nanoparticle fractions were determined by an ultraviolet-visible spectrophotometer. Harvested coffee edible plant-derived exosome-like nanoparticles were characterized by transmission electron microscopy. The quantification procedure was performed using a commercial kit. Coffee edible plant-derived exosome-like nanoparticles' proliferative effects on human hepatic stellate cells and human hepatocellular carcinoma cells were studied using an MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. Whole-exosome RNA sequencing was performed. RESULTS: Transmission electron microscopy scanning analysis indicated round-shaped nanoparticles with sizes ranging from 40 to 100 nm. Both size exclusion chromatography and kit-isolated edible plant-derived exosome-like nanoparticle samples showed maximum absorbance at 227.5 nm in ultraviolet-visible spectrophotometer analysis. Regarding the quantitation results, kit isolation was more efficient than the size exclusion chromatography method when the harvested particle numbers were compared. An important MTT assay finding confirmed the observed beneficial effects of coffee beverages: coffee edible plant-derived exosome-like nanoparticles significantly suppressed hepatocellular carcinoma cell proliferation. As a result of sequencing, we identified 15 mature miRNAs. A MapReduce-based MicroRNA Target Prediction Method (The DIANA tools' MR-microT algorithm) highlighted 2 genes specifically associated with the miRNAs that we obtained: KMT2C and ZNF773. CONCLUSION: For the first time in the literature, coffee edible plant-derived exosome-like nanoparticles were identified. These nanoparticles may have therapeutic effects on chronic liver diseases. Experimental studies, therefore, should be performed on disease models to demonstrate their efficacy.

The cost and profitability analysis of different dairy farm sizes.

Dairy cattle production is a substantial industry in Türkiye's livestock sector, with a significant economic impact. Because of its proximity to Türkiye's major market zone, the Mediterranean and Aegean regions, the province of Isparta can develop its milk production capacity and make investments in dairy production. Therefore, the purpose of this study was to look into production costs and drivers of profitability for dairy farms of various sizes in Isparta, a major milk-producing region in Türkiye. The study included data from 159 dairy cattle farms operating in the production period 2020-2021 in the designated area. Dairy farms were separated into three groups based on size and analysed consequently. The findings revealed that total feed consumption was higher on small-scale farms, while milk production was lower. Feed costs were the highest portion of the total production cost among the cost items (72.86%), followed by permanent labour costs (7.12%). Furthermore, milk sales income (64.39%) was the largest contributor to the average income in terms of the gross product value. Aside from milk production, it was discovered that cattle value appreciation (24.12%) increased farm income. Consequently, as farm size increased, production costs per animal unit fell while net profit rose. Finally, feed is the most significant input that boosts milk production costs. Also, larger farms were found to be more profitable in the study area. Thus, it was concluded that policies that could have a favourable effect on an increase in the cattle population on the farm should be advanced in the study area.

Method validation for the determination of lead in raw cow's milk by electrothermal atomic absorption spectrometry.

In this study, lead in raw cow's milk has been determined by validated electrothermal atomic absorption spectrometry (ETAAS) with Zeeman-effect background correction. Maximum pyrolysis and optimum atomization temperatures of lead were determined in the presence of modifiers. Pd + Mg(NO3)2 has been found a powerful modifier mixture for the determination of lead. The analytical parameters of the method such as limit of detection, limit of quantification and the effect of interfering ions have been investigated. The detection limit (3sigma) achieved by the method was calculated to be 0.62 ng/mL for Pb. Repeatability of the method evaluated as the relative standard deviation of 16-17 replicates using 5 ng/mL, under optimum experimental conditions were about 1.5% for synthetic sample solution and about 3% for real sample (N = 5). The described method has been validated by analyzing certified reference material (BCR-CRM 150) and by comparing the results with those obtained by inductively coupled plasma mass spectrometry (ICP-MS). The validated method was applied to raw cow's milk samples produced in 7 different regions of Turkey in 2003-2004. Raw cow's milk contained a mean (range) of 31.4 (2.5 - 313) microg kg(-1) lead with a relative error below 2%.

On-line speciation and determination of Cr(III) and Cr(VI) in drinking and waste water samples by reversed-phase high performance liquid chromatography coupled with atomic absorption spectrometry.

A simple, rapid, and selective on-line method for the speciation and determination of Cr(III) and Cr(VI) in aqueous solutions by ion-pairing HPLC coupled with flame atomic absorption spectrometry (FAAS) is described. The composition of the mobile phase has been optimized for better separation. The effects of column temperature, volume of injection loop, fuel flow rate of FAAS, and nebulizer suction rate of FAAS have also been investigated. Separation is accomplished in almost 2.5 min on a 25 cm length C18 column at 40 degrees C. The selectivity of the method has been established by investigating the effect of interfering elements on chromium determination. The detection limit (3sigma) achieved by the method was calculated as 3.7 ng/mL for Cr(III) and 2.0 ng/mL for Cr(VI). The proposed method has been validated by analyzing certified reference material (BCR 544) and successfully applied to the analysis of drinking water and wastewater samples with a relative error below 6%.