Human Blood Lymphocytes Research Articles

Effects of 660 nm red light and 850 nm near infrared light on human blood lymphocytes

Abstract The applications of light emitting diodes (LED) as a red light therapy (RLT) source for the management of various conditions such as wound treatment, control of inflammation and pain have been growing continually. Despite its extensive medical application, the effects of RLT on living cells are still highly contested. This study is conducted to test the effects of RLT using combined LED source with 660 nm (red) and 850 nm (near infrared) light on cultured human lymphocytes in vitro. To analyze the effect of RLT on human peripheral blood lymphocyte’s proliferation, the mitotic index (cytostatic effect) is monitored under different irradiation exposure time parameters and conditions. This value indicates how a particular treatment affected cell division, either proliferatively, inhibitory, or had no effect at all. The interpretation of the results of the mitotic index is done in relation to the mitotic index of the untreated (unirradiated) control cells. Our data shows higher lymphocyte proliferation for all of the irradiated samples, and is particularly enhanced by multiple exposures to red light. The effectiveness of RLT on cell activity is of importance in determination of suitable treatment for diseases related to the immune system. To better understand the molecular and metabolic mechanisms involved in red LED-induced photobiomodulation, the study will be extended to investigate the RLT effect on cell protein synthesis.

Vitamin B12 Protects Against Genotoxicity Induced by Cisplatin.

Cisplatin is an effective synthetic chemotherapeutic drug used for cancer treatment. Vitamin B12 has been shown to possess anti-genotoxic activity. This study aimed to investigate the effect of vitamin B12 on chromosomal damage induced by cisplatin. The level of sister chromatid exchanges (SCEs) and chromosomal aberrations (CAs) were measured in cultured human blood lymphocytes treated with cisplatin and/or vitamin B12. The results showed a significantly elevated frequency of CAs and SCEs of cisplatin-treated cultures compared to the control (P < 0.05). The CAs and SCEs induced by cisplatin were significantly lowered by pretreatment of cell cultures with vitamin B12. In addition, cisplatin caused a slight reduction in the mitotic index (MI), while vitamin B12 did not modulate the effect of cisplatin on MI. Vitamin B12 can protect human lymphocytes against genotoxicity associated with cisplatin.

Cytogenetic Violations in Blood Lymphocytes of Macaca Mulatta Monkeys in the Long Term after Irradiation with Accelerated Krypton Ions

A study has been performed of cytogenetic violations that occur in blood lymphocytes of Macaca mulatta monkeys after local exposure of the hippocampal region to accelerated 78Kr ions at a dose of 3 Gy. Analysis revealed a low level of chromosomal aberrations in the control group of monkeys. The number of cells with chromosomal aberrations did not exceed 1.67٪. The main fraction was chromatid-type aberrations. A cytogenetic analysis of peripheral blood lymphocytes of monkeys exposed to accelerated krypton ions revealed the maximum level of chromosomal aberrations 24 h after exposure, which exceeded the control level by 2.7 times. By day 96, the number of aberrations decreased 1.7-fold. However, it still exceeded the control level. By 460 days, some animals showed a slight increase in the number of aberrations compared to the previous observation period. In the long term, a number of irradiated animals showed deviations from the standard behavior of monkeys. Data on chromosomal aberrations in blood lymphocytes of monkeys and humans after in vitro irradiation with 170 MeV protons are compared. The relevance of this research is due to the preparation of interplanetary manned flights.

Toxicity of Dibutyl phthalate (DBP) toward isolated human blood lymphocytes: Apoptosis initiated from intracellular calcium enhancement and mitochondrial/lysosomal cross talk

Dibutyl phthalate (DBP) is a phthalate ester with wide application in industrial products, so human exposure can happen in workplaces and environment. Conflicting results have been acquired in researches which measured the influences of phthalates contact on immune responses in laboratory animals. Nevertheless, the straight influence of DBP on human lymphocytes and entire mechanisms of its effect against these cells continue to be unexplored. The major purpose of present research was to evaluate the mechanisms which lead to the DBP toxicity on human lymphocytes using accelerated cytotoxicity mechanisms screening (ACMS) technique. Cell viability was determined following12h incubation of lymphocytes with 0.05–1 mM DBP, and mechanistic parameters were assessed after 2, 4 and 6 h of lymphocyte treatment with ½ the IC5012h (0.3 mM), the IC5012h (0.6 mM) and twice the IC5012h (1.2 mM) of DBP. The IC5012 h of a chemical/toxicant is defined as concentration that kills 50 % of cells after 12 h of exposure. The results indicate that DBP exerts toxic effects on isolated human lymphocytes, probably through mitochondrial and lysosomal damage induced by glutathione depletion and oxidative stress. In this study, suppression of cytokines (IL2, INF-gamma and TNF-alpha) production and increase in intracellular calcium were also related to DBP induced lymphocyte toxicity.

Effects of Rubus idaeus extracts on some Cancerous Cell Lines in vitro Study Using MTT Assay

Our study investigated the cytotoxicity of Rubus idaeus alcoholic and aqueous extracts prepared from ripe blackberries were collected from the farmlands north of Baghdad, the extracts prepared at the same starting concentration, and tested on two cell line, the result show high toxic effects increase with the increase of concentration. The three extracted components from Rubus idaeus (purified flavonoid, polysaccharides, and carotenes) showed cytotoxic effect towards both: the primary cell culture of normal hepatic cells (HepG2 and L20B cell), and cancer hepatic cell lines (HepG2 and L20B cell) at 100μg/ml concentration for 24 hours treatment. Purified flavonoid exerted the potent effect on both cell lines among the other two extracted components. The cytotoxicity assay was held only for the purified flavonoid to investigate the mechanism by which the purified flavonoid affected living cells toward apoptosis. The most significant reduction (p≤0.05) in cell viable count was at the concentration 100μg/ml which appear to cause the induction of cell death via mitochondrial pathway for HepG2 and L20B cell line after 24 hours exposure. The purified flavonoid had no effect on the HepG-2 cell cycle. The Immunomodulation effect for the purified flavonoid and the extracted polysaccharides on normal human peripheral blood lymphocytes showed that the purified flavonoid suppress lymphocytes proliferation, while the later increased lymphocytes proliferation significantly. The immune stimulating effect of the polysaccharides caused alteration in IL-17 and level estimated by ELISA technique towards IL-2 elevation for the normal human blood lymphocytes against IL-17 level after 4 hours exposure at concentrations (250 and 500 μg/ml), while toxicity results were shown after 4 hours.

Impact of Ionic Liquids on the Physicochemical Behavior of Vesicles.

The effects of two ionic liquids (ILs), 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim]BF4) and 1-butyl-1-methyl pyrrolidinium tetrafluoroborate ([bmp]BF4), on a mixture of phospholipids (PLs) 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), and 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) (6:3:1, M/M/M, 70% PL) in combination with 30 mol % cholesterol (CHOL) were investigated in the form of a solvent-spread monolayer and bilayer (vesicle). Surface pressure (π)-area (A) isotherm studies, using a Langmuir surface balance, revealed the formation of an expanded monolayer, while the cationic moiety of the IL molecules could electrostatically and hydrophobically bind to the PLs on the palisade layer. Turbidity, dynamic light scattering (size, ζ-potential, and polydispersity index), electron microscopy, small-angle X-ray/neutron scattering, fluorescence spectroscopy, and differential scanning calorimetric studies were carried out to evaluate the effects of IL on the structural organization of bilayer in the vesicles. The ILs could induce vesicle aggregation by acting as a "glue" at lower concentrations (<1.5 mM), while at higher concentrations, the ILs disrupt the bilayer structure. Besides, ILs could result in the thinning of the bilayer, evidenced from the scattering studies. Steady-state fluorescence anisotropy and lifetime studies suggest asymmetric insertion of ILs into the lipid bilayer. MTT assay using human blood lymphocytes indicates the safe application of vesicles in the presence of ILs, with a minimal toxicity of up to 2.5 mM IL in the dispersion. These results are proposed to have applications in the field of drug delivery systems with benign environmental impact.

Effects of Partial-Body, Continuous/Pulse Irradiation at Dose Rates from FLASH to Conventional Rates on the Level of Surviving Blood Lymphocytes: Modeling Approach II. Two- and Multiple-Pulse Irradiation.

Mathematical models, which describe effects of partial-body, two- and multiple-pulse irradiation at high total doses D and at average dose rates N from FLASH to conventional rates on the level of surviving blood lymphocytes in humans and mice, have been developed originating in the previously proposed approach. These models predict that levels of surviving blood lymphocytes in humans and mice increase with increasing the dose rate from N=D/TR (TR is the time of the blood flowing into or out of the irradiated segment of the blood circulatory system) to FLASH rates and approach an upper limiting level equal to (1-vR), where vR is the fraction of blood volume in the irradiated segment of the blood circulatory system. Levels of surviving blood lymphocytes computed at total doses D of 10-40 Gy and at average of dose rates N, which are equal to or exceed 40 Gy/s for humans and 400 Gy/s for mice, are nearly indistinguishable from the upper limiting level. These results can be interpreted as the models reproducing the optimal blood lymphocyte sparing in these mammals after such exposures. With decreasing the dose rate from N=D/TR to conventional rates, at multiple-pulse irradiation the levels of surviving blood lymphocytes in humans and mice decrease to lower limiting levels, whereas at two-pulse irradiation they change cyclically and do not fall below their values for the delivery time equal to TR. Additionally, effects of two- and multiple-pulse irradiation of the whole abdomen in mice on the level of surviving blood lymphocytes are simulated within the developed models. Regimens of two- and multiple-pulse irradiation are taken the same as those reported in experiments, where effects of such exposures on the level of surviving crypts in mice were studied. Juxtaposing the modeling results with the experimental data reveals that the level of surviving blood lymphocytes in mice after two- and multiple-pulse irradiation of the abdomen at average dose rates N from FLASH to conventional rates modulates the level of surviving crypts in these animals after such exposures. A hypothesis is proposed to explain this phenomenon.

Effects of Partial-Body, Continuous/Pulse Irradiation at Dose Rates from FLASH to Conventional Rates on the Level of Surviving Blood Lymphocytes: Modeling Approach. I. Continuous Irradiation.

A mathematical model developed by Cucinotta and Smirnova is extended to describe effects of continuous, partial-body irradiation at high doses D and at dose rates N from FLASH to conventional rates on the level of surviving blood lymphocytes in humans and small laboratory animals (mice). Specifically, whereas the applicability of the model is limited to the exposure times shorter than a single cardiac cycle T0, the extended model is capable of describing such effects for the aforementioned and longer exposure times. The extended model is implemented as the algebraic equations. It predicts that the level of surviving blood lymphocytes in humans and mice increases with increasing the dose rate from N= D/T0 to FLASH rates and approaches the upper limiting level of 1-vR, where vR is the fraction of blood volume in the irradiated part of the blood circulatory system. Levels of surviving blood lymphocytes computed at doses from 10 Gy to 40 Gy and at dose rates N, which equal or exceed 40 Gy/s for humans and 400 Gy/s for mice, are nearly indistinguishable from the upper limiting level. In turn, the level of surviving blood lymphocytes in humans and mice decreases with decreasing the dose rate from N= D/T0 to conventional rates and approaches a lower limiting level. This level strongly depends on the dose D (it is smaller at larger values of D) with a slight dependence on the dose rate N. The model with the parameters specified for mice (together with the model of the dynamics of lymphopoietic system in mice after partial-body irradiation) reproduce, on a quantitative level, the experimental data, according to which the concentration of blood lymphocytes measured in mice in one day after continuous, partial-body irradiation at a high dose and conventional dose rate is smaller at the larger value of vR. Additionally, the model predicts at the same high dose (>10 Gy) a faster restoration of the blood lymphocyte population in humans exposed to continuous, partial-body irradiation at a FLASH dose rate compared to a conventional dose rate.

RABiT-III: an Automated Micronucleus Assay at a Non-Specialized Biodosimetry Facility.

Micronuclei, detected through the cytokinesis-block micronucleus assay, are valuable indicators of ionizing radiation exposure, especially in short-term lymphocyte cultures. The peripheral human blood lymphocyte assay is recognized as a prime candidate for automated biodosimetry. In a prior project at the Columbia University Center for Radiological Research, we automated this assay using the 96-well ANSI/SLAS microplate standard format and relied on established biotech robotic systems named Rapid Automated Biodosimetry Tool (RABiT). In this study, we present the application of a similar automated biotech setup at an external high-throughput facility (RABiT-III) to implement the same automated cytokinesis-block micronucleus assay. Specifically, we employed the Agilent BRAVO liquid-handling system and GE IN Cell Analyzer 6000 imaging system in conjunction with the PerkinElmer Columbus image data storage and analysis system. Notably, this analysis system features an embedded PhenoLOGIC machine learning module, simplifying the creation of cell classification algorithms for CBMN assay image analysis and enabling the generation of radiation dose-response curves. This investigation underscores the adaptability of the RABiT-II CBMN protocol to diverse RABiT-III biotech robotic platforms in non-specialized biodosimetry centers. Furthermore, it highlights the advantages of machine learning in rapidly developing algorithms crucial for the high-throughput automated analysis of RABiT-III images.

HPLC analysis, genotoxic and antioxidant potential of Achillea millefolium L. and Chaerophyllum villosum Wall ex. Dc

BackgroundMethanolic and chloroformic extract of Achillea millefolium and Chaerophyllum villosum were evaluated for HPLC analysis, genotoxic and antioxidant potential.Materials and methodsGenotoxic activity was carried out on human blood lymphocytes via comet assay and antioxidant activity was studied through DPPH method.ResultsThe genotoxic potential of A. millefolium and C. villosum’s methanolic and chloroformic extract was analysed using comet assay technique. Comet shaped human lymphocytes cells were observed when treated with different concentrations (50 mg/mL, 75 mg/mL, 100 mg/mL) of methanolic and chloroformic extract of both plants. Reading was taken on the basis of damaged DNA head and tail length. Greater the length of tail as compared to head, greater will be the damage and vice versa. Total comet score was obtained from A. millefolium subjected to different concentrations. After a time interval of 24 h both the extract showed dose dependant genoprotection with maximum genoprotectivity at 98.7 ± 12.7 and 116 ± 5.3 at 50 mg/100 mL for methanolic and chloroformic extract respectively. Similarly Total Comet score was obtained from C. villosum subjected to different concentrations of methanolic and chloroformic extract. After 24 h exhibited dose dependent genoprotection with maximum protectivity at 85.7 ± 22.0 and 101.7 ± 8.6 at 50 mg/100 mL for methanolic and chloroformic extract were determined. The antioxidant activity revealed that methanolic extract of A. millefolium showed highest antioxidant activity (84.21%) at 300 mg/ml after 90 min while the chloroformic extract of C. villosum exhibited highest (68.46%) antioxidant activity (59.69%) at 300 µg/ml after 90 min but less than the standard drug ascorbic acid (88.72%). Quantitative phytochemical screening revealed high percentage of alkaloids (27.4%), Phenols (34.5%), Flavonoids (32.4%) as compared to Tannins (12%) in methanolic extract of A.millefolium. While high percentage of alkaloids (31.4), Phenols (19.3%), Flavonoids (35.5%) as compared to Tannins (16.6%) in chloroformic extract of C. villosum.ConclusionThe present results showed that A. millefolium and C. villosum possess a number of important compounds and revealed genoprotective property which may be used to treat several genetic disorders such as alzeimer’s disease in future (Grodzicki W, Dziendzikowska K, Antioxidants 9(3):229, 2020).

Effects of genome instability under irradiation in different CT scanning modes. Results of ex vivo pilot cohort study

Introduction. Medical radiation is one of the leading sources of public exposure in the world. In recent decades, the total number of X-ray diagnostic procedures has increased significantly, and with the increase in the volume of computed tomography (CT), a significant gain in the total cumulative radiation dose is also associated. &#x0D; The aim of the work is to compare the genotoxic effects of irradiation of human blood lymphocytes using various CT protocols. &#x0D; Materials and methods. Among patients of different genders and ages who sought for a preventive examination, nine practically healthy volunteers (donors) who signed an informed consent, were randomly selected to participate in the ex vivo experiment. 4 venous blood samples from each donor were irradiated on various CT protocols (0.82–11.8 mSv) in an anthropometric phantom of the human chest. Aliquots of each sample were cultured under conditions of cytokinesis block, recorded and analyzed according to the protocol of cytomic analysis in a micronucleus test.&#x0D; Results. Irradiation of blood samples in the Ultra-NDCT mode (0.82 mSv) revealed a peak in the proliferation of rapidly dividing cells and the frequency of genetic damage in them, and also demonstrated a high probability of the formation and consolidation of genetic damage in generations of dividing cells. This indicates increased genotoxicity and, most likely, immunotropicity of the studied irradiation mode.&#x0D; Llimitation of the study is the inadmissibility of involving a person in an experiment or using biomaterials without obtaining his consent, as well as the inadmissibility of causing physical harm or harm to human honor and dignity during the experiment&#x0D; Conclusion. When choosing CT modes, it is necessary to take into account not only the levels of effective doses, but also the possibility of developing the effects &#x0D; of genome instability. However, this approach requires additional genotoxic studies of CT protocols in the range from &lt;1 to 100 mSv.

Assessing the Association of the Degree of DNA Methylation and the Frequency of Chromosomal Aberrations in Human Lymphocytes in a Single Irradiation of Blood &lt;i&gt;in vitro&lt;/i&gt;

The most sensitive biomolecule under radiation exposure is DNA, whose damage manifests itself in the form of chromosomal aberrations (CA). The processes of DNA methylation, which are involved in the regulation of gene expression, replication, DNA repair, etc., are also affected by gamma radiation. The aim of the study was to evaluate the relationship between the degree of DNA methylation and the frequency of CA after acute in vitro irradiation of human blood lymphocytes with gamma radiation. The study involved 10 conditionally healthy workers of the Siberian Chemical Combine, in whose blood lymphocytes the degree of methylation of CpG-dinucleotides (wide-genome bisulfite sequencing, XmaI-Reduced representation bisulfite sequencing – XmaI-RRBS) and the frequency of CA (cytogenetic study) after acute in vitro blood irradiation with doses of 0 and 1.5 Gy were evaluated. After acute exposure to gamma radiation in lymphocytes, the frequency of aberrant cells, dicentric chromosomes, chromatid and chromosomal fragments increased. Correlation analysis of the status of CpG-dinucleotide methylation and the frequency of CA revealed changes in the degree of methylation of 97 genes, which strongly correlated positively (56 genes) or negatively (41 genes) with an increased frequency of CA. A primary genome-wide screening of genes whose methylation is correlates with a high frequency of CA was carried out. Many of the identified genes are promising as potential markers of radiation exposure and to study the mechanisms of formation of radiosensitivity of the body and radioresistance of tumors during radiation therapy.

Влияние внутриклеточной регуляции метаболизма на популяционный состав лимфоцитов периферической крови человека

Metabolic activity has a significant impact on the differentiation, proliferation and functioning of T cells. Different lymphocyte subpopulations use, to varying degrees, glycolysis and mitochondrial metabolism, whose main regulators are hypoxia-inducible factor 1-alpha (HIF-1α) and sirtuin 3 (SIRT3), respectively. The purpose of this paper was to study changes in the population composition of peripheral blood lymphocytes in humans depending on the level of the intracellular metabolic regulators SIRT3 and HIF-1α. Materials and methods. 227 residents of the city of Arkhangelsk and the Arkhangelsk Region were examined (mean age 42 ± 11 years). Absolute lymphocyte count was determined using the Sysmex XS 500i haematology analyser, while CD3+, CD4+, CD8+, CD10+, CD25+ and CD95+ phenotypes content, by indirect immunoperoxidase reaction. Intracellular adenosine triphosphate (ATP) concentration was measured using the luciferase bioluminescence method. HIF-1α and SIRT3 concentrations were measured in lymphocyte lysate using enzyme immunoassay. To divide the total sample into groups according to SIRT3 and HIF-1α content, k-means clustering was utilized. Results. Changes in SIRT3 and HIF-1α intracellular concentrations correlated with ATP concentration. It was found that in the group with high HIF-1α content, the proportion of CD4+, CD8+, CD10+ and CD25+ lymphocytes was greater than in the group with high SIRT3 concentration, which had a greater proportion of CD95+ lymphocytes. Thus, the content of intracellular metabolic regulators that regulate ATP production pathways in the cell, i.e. oxidative phosphorylation (SIRT3) and glycolysis (HIF-1α), affects the population composition of lymphocytes and is therefore important for assessing the immune response.

Strawberry (Fragaria × ananassa) and Kiwifruit (Actinidia deliciosa) Extracts as Potential Radioprotective Agents: Relation to Their Phytochemical Composition and Antioxidant Capacity

Ionising radiation is an important form of treatment for human cancer; however, the side effects associated with oxidative damage caused by radiation compromise its effectiveness. This work aimed to quantify the major bioactive components of freeze-dried kiwifruit (KD) and strawberry (SD) extracts and assess their potential efficacy as radioprotective agents in human blood lymphocytes. Their possible genotoxic and cytotoxic effects were also evaluated. The study was conducted by pre-treating human lymphocytes with KD and SD (50, 400, and 800 µg/mL) before radiation at 2 Gy. The results showed that SD presented a higher antioxidant capacity (12.6 mmol Trolox equivalents/100 g db) and higher values of total phenolic compounds (2435 mg of gallic acid equivalents/100 g db), while KD had the highest vitamin C content (322 mg ascorbic acid/100 g db). Regarding phenolic compounds, pelargonidin-3-glucoside was the most abundant in SD (1439 mg/1000 g db) and quercetin-3-O-galactoside in KD (635 mg/1000 g db). None of the tested concentrations of both fruit extracts showed a genotoxic effect. SD (800 µg/mL) reduced the sister chromatid exchange frequency and mitotic index. The efficacy of KD (400 and 800 µg/mL) in lowering the dicentric chromosome frequency demonstrated its radioprotective activity.

Mitochondrial Transplantation Attenuates Toxicity in Human Lymphocytes Caused by Clozapine and Risperidone

Background: Clozapine (CLZ) and risperidone (RIS) are drugs that have the ability to disrupt mitochondrial function. Also, these drugs increase the level of free radicals. Mitochondrial dysfunction plays a role in the etiology of various diseases. Replacement and treatment of defective mitochondria with healthy mitochondria have been considered. Mitochondrial therapy (mitotherapy) or exogenous mitochondria transplantation is a method that can be used to replace dysfunctional mitochondria with healthy mitochondria. This method can help in the treatment of diseases related to mitochondria. Methods: In this study, we investigated the transplantation effect of isolated lymphocyte mitochondria on the toxicity induced by CLZ and RIS on human blood lymphocytes. Lymphocytes were isolated using the Ficoll standard method. Mitochondria of human lymphocytes were used for mitotherapy. This study was conducted in 6 groups. After treatment, the level of reactive oxygen species (ROS), mitochondrial membrane potential (MMP), reduced glutathione (GSH) content, oxidized glutathione (GSSG) content, and adenosine triphosphate (ATP) content were evaluated. Results: Our data showed that CLZ (70 µm) and RIS (24 nM) caused cytotoxicity on human blood lymphocytes which are associated with ROS generation, collapse in MMP, decrease in GSH content, increase in GSSG content and change in ATP content. Mitochondria transplantation results showed that adding mitochondria of lymphocytes could protect the lymphocytes against the toxicity effects caused by CLZ and RIS. Furthermore, the results showed that pre-incubation with cytochalasin D considerably reserved the protective effects of mitotherapy in the human lymphocytes. Conclusion: We proposed that mitochondria transplantation or mitotherapy-affected blood lymphocytes with exogenous mitochondria could be used to treat CLZ and RIS-induced toxicity.

Cytotoxicity against human cancer cells and lymphocytes based on cerium oxide nanostructures

Nanomedicine has emerged as a promising avenue for targeted cancer therapy, aiming to develop innovative approaches with improved efficacy and reduced side effects. CeO2 Nanostructures (CeO2 NSs) have attracted considerable attention in biomedical research due to their unique physicochemical properties and potential applications in cancer therapy. In the present article, we studied various methods of synthesis of biocompatible cerium oxide nanoparticles through phyto synthesis (CeO2-P) using Heritiera fomes plant extract and the chemical reducing method (CeO2-C). As obtained, CeO2-P and CeO2-C were characterized using XRD (X-ray diffraction), FTIR(Fourier Transformation Infrared spectroscopy), UV-Visible spectroscopy, and field emission scanning electron microscopy. Further, cytotoxic activity was studied on human cancer cell lines – MDA-MB-231-human breast cancer cell line; HeLa -human cervical cancer cell line; HCT-116- human colon cancer cell line, and human blood mononuclear cells (lymphocytes). A comparative study evaluating the induction of apoptosis by CeO2-P and CeO2-C was performed using propidium iodide staining on the mentioned human cancer cell lines. Toxicity assessment results showed the better performance of CeO2-P nanoparticles against HCT-116 at an IC50 concentration of 6.37 μg/mL compared to CeO2-C nanoparticles (HCT-116 - 8.08μg/mL). Similarly, the CeO2 nanoparticles showed significant cytotoxicity against Hela and MDA-MB-231 cells after 24 h treatments. Interestingly, CeO2-P nanoparticles are less toxic to human blood lymphocytes than CeO2-C nanoparticles. Furthermore, propidium iodide staining revealed the signs of apoptosis in all three human cancer cell lines treated with CeO2-P compared to CeO2-C nanoparticles. In conclusion, these findings provide valuable insights into the potential use of CeO2 nanostructures as a promising therapeutic agent in cancer treatment while exhibiting minimal toxicity towards lymphocytes. Further investigations are warranted to explore the underlying mechanisms and optimize the therapeutic efficacy of CeO2 Nanostructures for targeted cancer therapy.

Intracellular Potassium in Cell Growth and Proliferation of Human Mesenchymal Stem Cells and Blood Lymphocytes

Many data show that K+ ions are essential for cell proliferation. In this brief review, we summarize our own studies and literature data that characterize the relationship between modulations of intracellular K+ content and the intensity of cell proliferation. Using flame emission assay we compared the transport of monovalent cations in transformed cells and in human mesenchymal stem cells in growing cultures, as well as during stress-induced cell cycle arrest and transition from monolayer (2D) to three-dimensional (3D) spheroids. A decrease in the intracellular content of K+ (evaluated as the ratio of the content of K+ in cells to the mass of cellular protein) associated with the accumulation of G1 cells in the population and accompanied by a stop in proliferation was revealed. The relationship between intracellular K+ and initiation of cell proliferation was further analyzed in human blood lymphocytes (HBL) as a model for the transition of cells from quiescence to proliferation. In HBL stimulated to proliferate, the content of K+ in cells increases during the transition G0/G1/S, accompanying the growth of small lymphocytes into blasts. Cellular water content, calculated from buoyant cell density, is higher in proliferating HBLs and in Jurkat leukemic T cells than in resting HBL. Available data suggest that intracellular K+ is important for successful cell proliferation as the main intracellular ion involved in the regulation of cell volume during the transition from quiescence to proliferation, and high K+ content and associated high water content in the cell are a characteristic feature of cell proliferation and transformation.

Molecular Immuno-response Effects for Iraqi Lycium barbarm Carotenes upon Normal Human lymphocytes culture

Objectives: The study aimed to investigate the effects of Iraqi Lycium barbarm Carotene on normal human blood lymphocytes at molecular level.&#x0D; Methods: The normal human blood lymphocytes culture was exposed to Carotene extracted from Iraqi Lycium barbarm to estimate the interleukins expression in treated lymphocytes represented by IL-10 and TNF-α as a moderator of immune response at a molecular scale, and their CD markers changes expression (CD3, CD4&amp;CD8) by flow-cytometer apparatus.&#x0D; Results: Total carotenes content was 0.33mg/g powdered dried fruit. Potent carotene effect was appeared at concentration of 500µg/ml for treated lymphocytes with increasing CD3 level to get upper level after 2 hours interval, while after 4 hours exposure both CD4 and CD8 levels increased dramatically for lymphocytes treated with 125 µg/ml, and 250µg/ml Lycium carotene. An alteration in cytokines gene expression for IL-10 production in response to carotene treatments at (125,250,500) µg/ml for both intervals, with suppress in tumor necrosis factor (TNF-α) gene expression in relation to β-actin the internal control and the PHA mitogenic agent.&#x0D; Conclusion: The study concluded that the carotenes extracted from Lycium barbarm fruits seemed to initiate the immune system toward Th2 cell activation rather thanTh1, besides the extract may potentiate IL-10 production.

Molecular design, synthesis and anticancer activity of new thiopyrano[2,3-d]thiazoles based on 5-hydroxy-1,4-naphthoquinone (juglone)

A series of 11-substituted 9-hydroxy-3,5,10,11-tetrahydro-2H-benzo[6,7]thiochromeno[2,3-d][1,3]thiazole-2,5,10-triones 3.1–3.13 were synthesized via hetero-Diels-Alder reaction of 5-ene-4-thioxo-2-thiazolidinones and 5-hydroxy-1,4-naphthoquinone (juglone). The structure of newly synthesized compounds was established by means of spectral data and a single-crystal X-ray diffraction analysis. The synthesized compounds were tested on a panel of cell lines representing different types of cancer as well as normal and pseudonormal cells and peripheral human blood lymphocytes. Compound 3.10 was found to be the most active derivative, exhibiting a cytotoxic effect similar to doxorubicin's one (IC50 ranged from 0.6 to 5.98 μM), but less toxic to normal and pseudonormal cells. All synthesized compounds were able to interact with DNA, although their anticancer activity did not correlate with the potency of interaction with DNA. The status of p53 in colorectal cancer cells correlated with the activity of the synthesized derivatives 3.1, 3.7, and 3.10. Compound 3.10 did not have an acute toxic effect on the body of С57BL/6 mice, unlike the well-known anticancer drug doxorubicin, which was used as a positive control. The injection of 3.10 (20 mg/kg) to mice had no effect on the counts of leukocytes, erythrocytes, platelets and hemoglobin level in their blood, in contrast to doxorubicin, which caused anemia and leukopenia, indicating bio-tolerance of 3.10in vivo.

Green synthesis of copper nanoparticles and their evaluation for antimicrobial activity and bio-compatibility

Developing antimicrobial resistance by microorganism is one of the major concerns of modern society. The emergence of antimicrobials resistance in bacteria against commonly used effective antibiotics results in the need for stronger and costly therapy. Nanoparticles seem to be a viable solution to tackle the problem of antibiotic resistance in bacteria. As the most of the nanoparticles was use more than one mechanism to kill microorganism. Here, the study was planned to synthesize plant extract mediated CuO nanoparticles with antimicrobial property using green synthesis method. The synthesized nanoparticles were characterized using the UV–vis spectrophotometer, Scanning electron microscope, and Fourier transfer infrared spectroscopy. Schefflera arboricola based synthesized nanoparticles were shows antimicrobial activity against gram negative (E. coli, P. aeruginosa) and Gram positive (B. subtilis,) group of bacteria. Human blood lymphocytes shows normal growth when in presence of Schefflera arboricola leaves extract based synthesized nanoparticles.