The importance of flowcytometry study with the first aspirate taken during bone marrow aspiration in the diagnosis of multiple myeloma and follow-up of minimal residual disease

Objective: Flow cytometry (FC) is a diagnostic method supporting traditional morphological examination in disease follow-up and the diagnosis of Multiple myeloma (MM). Normal and atypical plasma cells (PCs) can be told apart from each other by means of FC method. The plasma cell rate is the highest in the blood obtained in the first aspirate during bone marrow aspiration in MM. Material and methods: A total of 60 patients that have been diagnosed with MM between 2018 and 2020, including 30 patients whom flow cytometry was studied with the first aspirate during bone marrow aspiration, and 30 patients whom FC was studied with the second aspirate were included in our study. The characteristics of the patients were analyzed retrospectively from their files. Results: The median ratio of plasma cells (PCs) detected by FC and bone marrow biopsy was 17,5% and 44%, respectively. While this rate was median 37,5% in patients that flow cytometric study was performed with the first aspirate, the rate was found to be median 7% in patients that FC was performed with the second sample. The PCs rates were statistically significantly higher with the flow cytometric study with the first aspirate than the second one (p=0.000). Conclusion: Flow cytometric study with the first aspirate during bone marrow aspiration in patients with MM is diagnostically important.

In vitro and in vivo studies of cytotoxic effects of FeSO4 nanoparticles

Background and aims: Using iron as a food additive usually causes undesirable sensory changes and side effects in humans. In this study, we made iron (Fe) nanoparticles (NPs) and studied the cytotoxicity of FeSO4 bulk and NPs on HT-29 cells and different doses of these particles on rat intestine. Methods: Particle size of nanoscale was achieved by mechanical technique. Iron particles were characterized using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The effect of iron particles with different concentrations (6.25, 3.125, and 1.57 mM/mL) on the colon cell line was performed using the MTT assay at 24, 48, and 72 hours. Apoptosis and necrosis of the cells were assessed using Annexin V-FITC staining and propidium iodide (PI) at 24 h. In an in vivo study, Taftoon bread was produced from fortified wheat flour with FeSO4 bulk and NPs, which are recommended in human diet (9, 18, and 27 mg of elemental iron/kg flour). Wistar rats were fed daily with fortified bread for 21 days and their colon and small intestine were then evaluated histopathologically. Statistical analyses were performed using SPSS 22.0 software by chi-square test. Results: The synthesized FeSO4 NPs were smaller than 100 nm, and they had more adverse effects on the viability of the HT-29 cells compared to the bulk- FeSO4 at 72 hours. Flow cytometric study showed that the early apoptosis of cells by the bulk form was more than the NPs, but at the low concentration (1.57 mM/mL), the NPs induced more necrosis than the bulk particles (P=0.063). The survival rate of cells facing all concentrations of NPs and bulk- FeSO4 decreased dose dependently (P=0.075). In vivo results revealed that there were no pathological changes in rats’ intestinal tissues. Conclusion: The bulk and NPs of iron have adverse effects on the HT-29 cells, but no histopathological changes were seen on rats’ intestinal cells.

Characterization of endosperm development in Guggul genotypes based on flow cytometry

A comprehensive investigation on guggul endosperm development pattern was done by studying fruit set development under bagged condition and checked their ploidy level through flow cytometry. The study was carried out in 59 accessions maintained in the field gene bank of DMAPR. The flowers were bagged to avoid pollen grain and studied the fruit development pattern. It was found that, 48 accessions showed fruit set under bagged condition whereas 11 accessions failed to set the fruits. In these 11 accessions, the flowers were allowed for pollination and developing fruits (30-40 Days After Anthesis [DAA]) were collected and flow cytometric study was carried out to confirm triple fusion of endosperm development if there any. From the study it was observed that these showed 2x: 4x ploidy level, 2x:4x: 8x along with 2x: 3x which confirmed that, this apomictic species retains a low frequency of sexual reproduction. Hence it can be concluded that, both sexual and apomictic pathways are still active in the species which can be used as fixation of heterosis towards crop breeding improvement programme.

A Flow Cytometric Study of Reagent Cells to Resolve ABO Typing Discrepancy

Objectives RBC alloantibodies can lead to ABO grouping discrepancies unrelated to A or B antigens or antibodies posing challenges in the blood bank testing. Routine blood bank testing and flow cytometry were used to immunophenotype reagent cells and elucidate the cause of ABO discrepancies in two patients. Methods ABO discrepancy was identified in two patients after transfusion with several units of RBCs. For both patients, the pretransfusion type and screen demonstrated blood group A. Eight and 16 days later, both patients showed an apparent antibody to reagent group A cells, which prompted additional study with patients’ samples and flow cytometric testing of commercial reagent cells. Results In both patients’ specimens, posttransfusion evaluation demonstrated an emerging antibody to the Kell antigen (K). The RBCs of both patients typed negative for K, and both were transfused with K-positive RBCs. Flow cytometric analysis of reagent RBCs demonstrated that five of seven lot numbers were positive for K. Conclusions Emerging anti-K antibody led to agglutination of the K-positive reagent A1 cells, highlighting the importance of considering RBC alloantibodies and the composition of reagent cells when interpreting cases with an apparent ABO grouping discrepancy.

FUNCTIONAL ACTIVITY OF HEMOCYTES OF PULMONARY MOLLUSCS

Hemocytes, the cell of the hemolymph, play a key role in the immune response of pulmonate molluscs to various pathogens including trematode infection. The number of hemocytes is known to increase after immunization but the mechanism of their multiplication remains debatable. In this work we studied the functional and proliferative activity of hemocytes in two species of pulmonate molluscs: Biomphalaria glabrata, Planorbarius corneus. ImageStream technique was used for the study of the hemocyte populations of these molluscan species for the first time. The hemocytes of all the studied species were represented by two main types, granular cells and hyalinocytes. Microscopic and flow-cytometric study of the hemocytes with the use of EdU revealed some EdU-positive cells. However, the analysis of the cell cycle of the hemocytes showed that the amount of DNA in these cells was not increased.