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.

Screening of Variations in CD22 Gene in Children with B-Precursor Acute Lymphoblastic Leukemia.

BACKGROUND: CD22 is expressed on the surface of B-cell lineage cells from the early progenitor stage of pro-B cell until terminal differentiation to mature B cells. It plays a role in signal transduction and as a regulator of B-cell receptor signaling in B-cell development. OBJECTIVES: We aimed to screen exons 9-14 of the CD22 gene, which is a mutational hot spot region in B-precursor acute lymphoblastic leukemia (pre-B ALL) patients, to find possible genetic variants that could play role in the pathogenesis of pre-B ALL in Turkish children. METHODS: This study included 109 Turkish children with pre-B ALL who were diagnosed at Losante Hospital for Children with Leukemia. Genomic DNA was extracted from both peripheral blood and bone marrow leukocytes. Gene amplification was performed with PCR, and all samples were screened for the variants by single strand conformation polymorphism. Samples showing band shifts were sequenced on an automated sequencer. RESULTS: In our patient group a total of 9 variants were identified in the CD22 gene by sequencing: a novel variant in intron 10 (T2199G); a missense variant in exon 12; 5 intronic variants between exon 12 and intron 13; a novel intronic variant (C2424T); and a synonymous in exon 13. Thirteen of 109 children (11.9%) carried the T2199G novel intronic variant located in intron 10, and 17 of 109 children (15.6%) carried the C2424T novel intronic variant. CONCLUSION: Novel variants in the CD22 gene in children with pre-B ALL in Turkey that are not present, in the Human Gene Mutation Database or NCBI SNP database, were found.

Factor V Leiden and Prothrombin 20210A Mutations among Turkish Pediatric Leukemia Patients.

This study was undertaken to determine the prevalence of the Factor V 1691 G-A and PT 20210 G-A mutations in Turkish children with leukemia. We genotyped 135 pediatric leukemia patients with for these mutations. Eleven (8%) of the 135 patients were heterozygous for the FV 1691 G-A mutation. Seven (5,1%) of the patients carried the PT 20210 G-A heterozygous mutation. Of the 135 patients, only three had thrombotic event, none of which had these two mutations, which is common in Turkish population. Our findings revealed a controversial compared to the previous reports, which needs further investigation.

Minor anomalies in children with hematological malignancies.

BACKGROUND: Despite the presence of reports on correlation between major congenital defects and cancer, very few studies have investigated the frequency of minor anomalies in childhood malignancy. The aim of this study was to determine the prevalence of minor anomalies in children with hematological malignancy. PROCEDURE: A total of 62 well-defined minor anomalies were determined in 109 patients. The patients were compared with age- and sex-matched healthy control subjects. RESULTS: The results of this study showed that, 64.22% of the patients and 26.6% of the controls had at least one minor anomaly. Among the minor anomalies detected, pigmented nevi and café-au-lait spots were significantly more frequent in the patients. The prevalence of minor anomalies in the patients was significantly higher than that of the controls in the present study. CONCLUSION: Our results contribute to the understanding of the role of genetic factors in childhood hematological malignancies. Future studies may be directed toward identifying the developmental pathways and the relevant genes that are involved in the overlap between childhood hematological malignancies and minor anomalies.

A prospective series of musculoskeletal system rehabilitation of arthropathic joints in young male hemophilic patients.

The study aims to evaluate prospective results of physical therapy interventions and rehabilitation of arthropathic joints in young male hemophilic patients. Given that the effectiveness of the therapy would be higher if therapy is provided at early ages, a series of males aged 18 years or younger were included in the study. The study is a descriptive case series of 31 hemophilic children who consecutively referred to the physical therapy and rehabilitation clinic from the pediatric hematology clinic of the same hospital over a period of 12 months. After getting their oral informed consent, all the patients underwent an intensive, 4-week clinical rehabilitation program, concomitant with factor replacement treatment. A total of 65 arthropathic joints were evaluated in the study and physical therapy interventions were applied 5 days a week for 1 h daily. The range of motion (ROM), pain, clinical evaluation and disability scores were compared and contrasted before and after the 4-week physiotherapy program. The ROM of all involved joints improved (at knee, ankle and elbow) and the scores of pain, clinical evaluation and disability revealed statistically significant improvement (P < 0.001). A multidisciplinary approach is important in the management of hemophilic arthropathy and appropriate physiotherapy combined with adequate replacement factor therapy promotes maintenance of the musculoskeletal function in male hemophilic patients.

All-trans-retinoic acid-induced myositis in a child with acute promyelocytic leukemia.

Anthracyclin-based regimens and all-transretinoic acid (ATRA, tretinoin) as differentiating agent are commonly utilized for the treatment of acute promylelocytic leukemia (APL). There are many adverse effects that may be seen during the use of ATRA in patients with APL. Of these, ATRA-induced myositis is rarely described in adults and rare in the children with APL. Herein, we report an 11-year-old girl with APL who developed ATRA-induced myositis during induction treatment.

Functional disability in children with hemophilic arthropathy.

Hemophilia is a coagulation disorder characterized by acute hemorrhages into the musculoskeletal system, leading eventually to arthropathy and disability. We investigated the functional loss, namely disability, in hemophiliacs. The clinical and radiological characteristics of joint involvement were also evaluated. There were 31 patients between the ages of 3 and 18 years and 65 involved joints. The knees were the most commonly affected joints followed by the elbows and ankles. There was a positive correlation between the radiological and clinical evaluation scores; however, the clinical evaluation score did not correlate with age. The radiological score increased in conjunction with increasing age of the patients. We observed a significant relationship between the disability score and the clinical evaluation and radiological scores. These observations suggest that hemophilia is a life-long condition, with a high potential for functional disability if not promptly and adequately controlled. The main principle in the treatment of hemophilic arthropathy is the restoration of the patient's lifestyle and mobility with a comprehensive multidisciplinary approach.

Immunological status of thalassemia syndrome.

A major cause of morbidity and mortality in thalassemic patients is infections, assumed to be the result of immunological changes. In this study immune functions of peripheral blood lymphocytes have been studied in 38 beta-thalassemia major, 12 beta-thalassemia trait, and 17 healthy children. Results show decrease in CD4+/CD8+ ratios in the beta-thalassemia major group and no difference according to absolute T-lymphocyte numbers and activated T-cell numbers. These results do not correlate with the tendency to infection. No significant difference was found in humoral immunity. The study of other factors in thalassemia is needed to detect those who are more susceptible to infections.