Mouse Marrow Cells Research Articles

Acute and genetic toxicity of GS-E3D, a new pectin lyase-modified red ginseng extract

Korean red ginseng and its extract have been used as traditional medicines and functional foods in countries worldwide. Pectin lyase-modified red ginseng extract (GS-E3D) was newly developed as a dietary supplement for obesity, diabetes-related renal dysfunction, etc. In this study, the safety of GS-E3D on acute toxicity and genotoxicity was evaluated. For acute study, Sprague-Dawley rats were administrated by oral gavage at a dose of 5000 mg/kg GS-E3D. To evaluate genotoxicity of GS-E3D, we conducted three-battery tests, which are Ames test using Escherichia coli (WP2uvrA pKM101) and Salmonella typhimurium strains (TA98, TA100, TA1535 and TA1537), chromosomal aberration test -using Chinese hamster lung cells, and micronucleus test using ICR mice. In acute toxicity studies, there were no dead animals or abnormal necropsy findings in the control group and GS-E3D (5000 mg/kg) treated group. GS-E3D did not induce mutagenicity in the bacterial test, chromosomal aberrations in Chinese hamster lung cells and micronuclei in bone marrow cells of mice. Conclusively, the approximate lethal dose of GS-E3D was greater than 5000 mg/kg bw and GS-E3D has no genotoxic potential in the three-battery tests on genotoxicity.

Comparative Behaviour of Allogeneic Spleen versus Bone Marrow Cells in Non-Conditioned Neonatal Mice: Insights into Robust Heart Transplant Tolerance

Purpose Understanding the in vivo/in situ behaviour of fully allogeneic spleen (SC) vs bone marrow cells (BMC) in neonatal mice is important for optimizing appropriate synergistic interactions between the two to induce robust transplant tolerance. Allo-SC and -BMC were therefore injected either as a mixture or separately into neonates; their trafficking, fates, and effects on GVHD, HVGD and tolerance compared. Methods C3H (H-2k) neonates were injected with adult B6 (H2-2b) GFP+ SC or BMC. Cell trafficking, interactions and fates were monitored by whole body/organ imaging and high-resolution microscopy. GVHD was defined by reduced mouse growth/viability, HVGD by donor cell fragmentation and tolerance by donor-type heart transplantation. Results Allo-SC inoculum contained more immune effector cells than allo-BMC as determined by flow cytometry (CD8 T, NK, CD4 T cells). Neonates injected with allo-SC developed aGVHD with diarrhea, severe reduction in growth and death by 14 days post-injection (n=7/7) whereas allo-BMC-injected mice developed aGVHD infrequently (n=1/8). Mice injected with the SC/BMC mixture showed intermediate disease progression (n=6/7). Whole body/organ imaging showed allo-SC and -BMC each trafficked to secondary lymphoid organs, became activated, proliferated and spread throughout the mice suggesting systemic inflammation. GFP signal was stronger in mice injected with allo-SC than -BMC. Proliferating allo-SC and -BMC both contained T cells, including CD8 T cells. GFP signal intensities in syngeneic-SC and -BMC-injected mice were low at day 6 and confined to lymphoid organs indicating proliferation due to lymphopenia was limited. Surviving allo-BMC-injected mice transplanted as adults (n=6) with donor-type hearts showed moderate (2-3) to low (≤1) allograft beat scores at 100 days post-transplant. A 100-day allograft (score=1) showed cellular infiltration consisting of macrophages and T cells including CD8 T cells. Conclusion Both injected allo-SC and -BMC trafficked to secondary lymphoid organs, proliferated and spread throughout neonatal mice; allo-SC consistently caused aGVHD whereas allo-BMC induced non-robust heart transplant tolerance. These promising results suggest that robust tolerance induction will require different optimization procedures for allo-SC and -BMC. Understanding the in vivo/in situ behaviour of fully allogeneic spleen (SC) vs bone marrow cells (BMC) in neonatal mice is important for optimizing appropriate synergistic interactions between the two to induce robust transplant tolerance. Allo-SC and -BMC were therefore injected either as a mixture or separately into neonates; their trafficking, fates, and effects on GVHD, HVGD and tolerance compared. C3H (H-2k) neonates were injected with adult B6 (H2-2b) GFP+ SC or BMC. Cell trafficking, interactions and fates were monitored by whole body/organ imaging and high-resolution microscopy. GVHD was defined by reduced mouse growth/viability, HVGD by donor cell fragmentation and tolerance by donor-type heart transplantation. Allo-SC inoculum contained more immune effector cells than allo-BMC as determined by flow cytometry (CD8 T, NK, CD4 T cells). Neonates injected with allo-SC developed aGVHD with diarrhea, severe reduction in growth and death by 14 days post-injection (n=7/7) whereas allo-BMC-injected mice developed aGVHD infrequently (n=1/8). Mice injected with the SC/BMC mixture showed intermediate disease progression (n=6/7). Whole body/organ imaging showed allo-SC and -BMC each trafficked to secondary lymphoid organs, became activated, proliferated and spread throughout the mice suggesting systemic inflammation. GFP signal was stronger in mice injected with allo-SC than -BMC. Proliferating allo-SC and -BMC both contained T cells, including CD8 T cells. GFP signal intensities in syngeneic-SC and -BMC-injected mice were low at day 6 and confined to lymphoid organs indicating proliferation due to lymphopenia was limited. Surviving allo-BMC-injected mice transplanted as adults (n=6) with donor-type hearts showed moderate (2-3) to low (≤1) allograft beat scores at 100 days post-transplant. A 100-day allograft (score=1) showed cellular infiltration consisting of macrophages and T cells including CD8 T cells. Both injected allo-SC and -BMC trafficked to secondary lymphoid organs, proliferated and spread throughout neonatal mice; allo-SC consistently caused aGVHD whereas allo-BMC induced non-robust heart transplant tolerance. These promising results suggest that robust tolerance induction will require different optimization procedures for allo-SC and -BMC.

High-fat diet disturbs lipid raft/TGF-\u03b2 signaling-mediated maintenance of hematopoietic stem cells in mouse bone marrow

Despite recent in vivo data demonstrating that high-fat diet (HFD)-induced obesity leads to major perturbations in murine hematopoietic stem cells (HSC), the direct role of a HFD is not yet completely understood. Here, we investigate the direct impact of a short-term HFD on HSC and hematopoiesis in C57BL/6J mice compared with standard diet-fed mice. We detect a loss of half of the most primitive HSC in the bone marrow (BM) cells of HFD-fed mice, which exhibit lower hematopoietic reconstitution potential after transplantation. Impaired maintenance of HSC is due to reduced dormancy after HFD feeding. We discover that a HFD disrupts the TGF-β receptor within lipid rafts, associated to impaired Smad2/3-dependent TGF-β signaling, as the main molecular mechanism of action. Finally, injecting HFD-fed mice with recombinant TGF-β1 avoids the loss of HSC and alteration of the BM’s ability to recover, underscoring the fact that a HFD affects TGF-β signaling on HSC.

Compatibility effects of ginseng and Ligustrum lucidum Ait herb pair on hematopoietic recovery in mice with cyclophosphamide-induced myelosuppression and its material basis

BackgroundGinseng (G) and Ligustrum lucidum Ait (LLA) are core traditional Chinese medicines in treating myelosuppression formula. The present study was designed to profile effect of G and LLA herb pair (G-LLA) on myelosuppressed mice. MethodsThe mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (Cy). Hematopoietic function of bone marrow was measured by hemopoietic progenitor cell culture and peripheral blood count, and serum hemopoietic factors were tested by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. HPLC was used to measure 20 potential chemical components related to myelosuppression, including ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1, Rg5, salidroside, and so on. ResultsG, LLA, and G-LLA improved the amount of peripheral blood cells and bone marrow cells of myelosuppressed mice (P < 0.01). They significantly increased the colony quantity of colony-forming unit–granulocyte macrophage, burst-forming unit−erythroid, colony-forming unit−erythroid, and colony-forming unit−megakaryocyte and amount of G2/M and S phase cells (P < 0.01). They also significantly decreased the amount of hematopoiesis-related cytokines (P < 0.01). The content of chemical components in G-LLA changed, and the change of rare saponin was the most obvious. ConclusionThese results show that G-LLA herb pair might produce synergistic or complementary compatibility effects on bone marrow suppression after chemotherapy. It suggests that the substance basis of G-LLA for treating bone marrow suppression may be effective chemical components.

STUDYING THE POSSIBLE MUTAGENIC PROPERTIES OF NEW MEDICINE ON THE BASIS OF COMPLEX LITHIUM CITRATE, ALUMINUM OXIDE AND POLYMETHILSILOXANE

Aim of the study was to investigate the possible mutagenic properties of a new drug based on a lithium-containing substance – a complex of lithium citrate, polymethylsiloxane and aluminum oxide. Material and methods . Methods for testing mutagenicity using chromosomal aberrations in the bone marrow cells of CBA mice and somatic recombination in Drosophila melanogaster were used. Results . It was shown that a single intragastric administration of drug at a dose of 5000 mg/kg and a fivefold course of administration at a dose of 400 mg/kg to CBA mice did not increase the level of cytogenetic disorders in bone marrow cells. The study of the lithium complex drug in a somatic mosaicism test revealed that the preparation at a dose of 2000 mg/kg does not increase the frequency of mutations in Drosophila melanogaster. Conclusion . A single intragastric administration of the studied drug at a dose of 5000 mg/kg and its course administration (400 mg/kg × 5) do not increase the level of cytogenetic disorders in the bone marrow cells of CBA mice. In the somatic recombination (mosaicism) test system on D. melanogaster, no increase in the appearance of mutant setae and spots on the body and head was observed when using yellow and singed markers. The results of the study indicate that the studied drug does not have mutagenic properties.

Empirical Study on the Therapeutic Effect of Astragalus Polysaccharides in a Mouse Model of Ulcerative Colitis

This study was performed to study the effects of astragalus polysaccharides (APS) in the treatment of ulcerative colitis (UC), and to explore whether myeloid-derived suppressor cells (MDSC) involve in this process. UC model was established by dextran sulfate sodium salt (DSS) inducement in mouse. Then the effects of APS on UC was evaluate at the levels of cytology and molecular biology: exploring the influence of APS on MDSC by analyzing the levels of MDSC before and after the treatments of APS; evaluating the therapeutic effects of APS on UC by pathologic histology. Data showed that the levels of MDSC in bone marrow, spleen, peripheral blood of UC mice were significantly decreased after intervention with APS, indicating APS inhibited the level of MDSC significantly. After depletion bone marrow cells in mice, we further found that the therapeutic effects of APS in MDSC- group was significantly reduced compared with MDSC+ group. It was discovered through pathological analysis that, compared with UC model group, APS intervention group mainly manifested as reduced infiltrating acute and chronic inflammatory cells, necrotic epithelial cells and epithelial ulceration. Therefore, APS could reduce the inflammatory cell infiltration in colonic tissues of UC model mice, repair the damaged colonic mucosa, and promote ulcer healing. In conclusion, APS has a potential application in the treatment of ulcerative colitis, and is dependent or partially dependent on MDSC to achieve this effects.

Genotoxicity, anti-melanoma and antioxidant activities of Hymenaea courbaril L. seed extract.

Hymenaea courbaril has been used to treat different diseases, although its properties are yet to be scientifically validated. The objective of this study was to determine the cytotoxicity, genotoxicity, antigenotoxicity and antioxidant potentials of hydroethanolic extract from H. courbaril seeds. Therefore, for the cytotoxicity test an anti-melanoma assay was performed in B16F10 strain cells. The genotoxicity and antigenotoxicity was evaluated in bone marrow cells (Permit number: 002/2010) of mice, the antioxidant activity was determined by the DPPH test and the total flavonoid content was also determined. The hydroethanolic extract showed antigenotoxic effect and antioxidant activity. It was verified that total flavonoid content was 442.25±18.03 mg RE/g dry extract. HPLC-PAD chromatogram revealed presence of flavones as majority compound in evaluated extract. The results allowed us to also infer that the hydroethanolic extract from seeds shows cytotoxic activity against B16F10 melanoma cells line and it has dose-and-time-dependency.

Differentiation of bone marrow cells in arthritic mice with decreased complement activity

Differentiation of bone marrow cells in arthritic mice with decreased complement activity

Inducible Correction of a RUNX1 Mutation of Human AML Causes a Switch of AML to B-ALL

Background: Point mutations of RUNX1 gene are frequently seen in myeloid malignancies such as myelodysplasia (MDS) and acute myeloid leukemia (AML), and are associated with unfavorable clinical outcomes. AML patients bearing RUNX1 mutations have a significantly lower complete remission (CR) rate compared with patients with wild-type RUNX1 (30% vs. 73%), whereas in contrast, the CR rate of ALL patients, in which RUNX1 is rarely mutated, are at 92%. It remains unclear whether RUNX1 mutations found in AML patients are causal to AML maintenance, and if RUNX1 mutations serve as a valid therapeutic target in the treatment of such therapy-resistant AML.Experimental Approach: The RUNX1 mutations are found frequently associated with MLL-partial tandem duplication (MLL-PTD) in AML. We have developed two complementary mouse models that express a MLL-PTDknock-in mutation together with a RUNX1 S291fsX300 mutation found in AML patients: in the first model RUNX1 S291fsX300 was introduced into the bone marrow cells of the MLL-PTD knock-in mice by retrovirus transduction, while in the second model the tetracycline-inducible RUNX1 S291fsX300 mutation was knock-in at the Collagen a1 locus and the mice was crossed to MLL-PTD knock-in. In the second model, the mutant RUNX1 protein is only expressed when the mice were fed with doxycycline containing food and the RUNX1 mutant is absent or “corrected” upon doxycycline withdrawn. BM cells of these mice, as well as WT and MLL-PTD controls, were transplanted to syngeneic mice and the recipients were tracked for disease development and progression at bi-monthly intervals. The doxycycline induction or withdraw was carried out after secondary transplant, and the expression of RUNX1 S291fsX300 was monitored by a built-in GFP reporter and verified by RT-PCR.Under tetracycline induction, the RUNX1 S291fsX300 and MLL-PTD double mutation bearing mice developed a spontaneously AML and had a survival time 6-10 months after transplantation, and they showed symptoms of MDS/AML or AML, including thrombocytopenia, anemia, leukocytosis, splenomegaly, abnormal BM and spleen cell morphologies. We transplanted the AML mouse bone marrow to secondary recipients, and observed them with or without continuing doxycycline induction. By using the O-propargyl-puromycin to track the newly synthesized proteins, we found that the RUNX1 mut-on and RUNX1 mut-off mice showed comparable protein synthesis rate at full-blown leukemia stage, indicating that the RUNX1 mutation does not play a restrictive role to protein synthesis in transformed leukemia cells. The c-Kit+ AML cells cultured from the RUNX1 mut-off mice showed reduced viability and significantly reduced CFU-C activities, which are accompanied by a lineage switch in a decrease in expression of Mac1 and an increase in expression of B220. In vivo, the RUNX1 mut-off mice showed a slightly prolonged survival compared with RUNX1 mut-on group (182 vs. 126 days, p=0.0036). An analysis of the lineage distribution in their peripheral blood and bone marrow revealed that the RUNX1 mut-off AML mice underwent a dramatic lineage switch from myelocytes to lymphocytes (Fig. 1): while the Mac1+Gr1+population remained high in the RUNX1 mut-on AML mice, turning off RUNX1 mut expression caused an increased B cell population in expense of the Mac1+Gr1+population. Pathological analysis also showed a drastic decrease of the myeloid marker MPO and increased lymphoid marker B220 in the RUNX1mu-off mice. Consistently, a RT-PCR test of the BM c-Kit+ cells further showed that upon turning off the RUNX1mut, critical B cell regulatory genes including TCF3, EBF1, PAX5, CD79A were significantly up regulated whereas myeloid regulatory genes such as PU.1 were downregulated. In addition, RUNX1 S291fsX300 expression in the AML cells confers a resistance to chemotherapy (Ara-C) treatment compared with that of WT RUNX1 expression, indicating that the RUNX1 mutation is responsible for the AML resistance to therapy.Conclusion:Although RUNX1 mutations found in AML patient may not be essential for the hyperproliferative and growth phenotype of the AML, they play a causal role in maintaining myeloid lineage restriction, expansion and chemo-resistance. Correction or removal of the RUNX1 mutation in AML cells leads to a disease switch from AML to B-ALL, effectively changing the difficult-to-treat drug-resistant AML to another disease with more favorable CR (B-ALL). [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Interleukin 4 Induces Macrophage-Mediated Phagocytosis of Leukemia Cells

Acute myeloid leukemia (AML) is an aggressive disease associated with poor prognosis, hence, new therapeutic strategies are needed. The AML cells reside in the bone marrow niche and depend on external signals such as cytokines for their growth and survival. In contrast to providing a supporting role, cytokines can act as negative regulators of AML cells. We previously identified that interleukin 4 (IL-4) induces apoptosis of AML cells in a Stat6-dependent manner and demonstrated strong anti-leukemic activity of IL-4 in a murine AML model driven by MLL-AF9 (Peña-Martínez et al., Leukemia 2017). Ectopic expression of IL-4 in AML cells resulted in prolonged survival in C57BL/6 mice, with mice almost devoid of leukemia cells in bone marrow and spleen upon the time of sacrifice. However, the IL-4-induced apoptosis of AML cells did not fully explain the strong antileukemic activity of IL-4 in vivo.To investigate whether the anti-leukemic effect of ectopic IL-4 expression in vivo was microenvironment dependent, we used IL-2 receptor gamma(-/-) immunodeficient mice, which lack a functional IL-4 receptor type I complex, as recipients for transplantation of murine MLL-AF9 AML cells. Notably, there was no antileukemic activity of IL-4 in these mice, suggesting that the in vivo anti-leukemic effect of IL-4 was predominantly microenvironment-dependent. To identify the cell population that was responsible for the antileukemic effect of IL-4 stimulation, we performed flow cytometry analysis of bone marrow and spleen cells in mice exposed to high IL-4 levels. We found that the anti-leukemic activity of IL-4 was associated with a significant expansion of F4/80+ macrophages in both the bone marrow and spleen, which was confirmed by immunohistochemistry.To determine whether macrophages were responsible for the anti-leukemic effect of IL-4, we depleted macrophages with clodronate liposomes in mice receiving IL-4-secreting AML cells. Depletion of macrophages partially rescued the anti-leukemic effect of IL-4, indicating that macrophages stimulated with IL-4 were responsible for the antileukemic activity. Consistent with these findings, in macrophage differentiation assays with either murine or human monocytes, IL-4 enhanced the phagocytosis of leukemia cells.To characterize the IL-4-induced macrophages, we performed RNA sequencing of F4/80+ macrophages from animals exposed to high IL-4 levels. Gene expression analysis revealed IL-4-induced M2 polarization of the macrophages, as key markers of M2 activation such as Arg1, Retnla and Chil3 were upregulated. Moreover, consistent with the in vitro findings, gene set enrichment analysis revealed an enrichment of phagocytosis signatures in the macrophages stimulated with IL-4.Strikingly, we found that exposure of AML cells to IL-4 resulted in strong upregulation of CD47, a ‘don't eat me signal’, which protects cells from macrophage-mediated phagocytosis. By CRISPR/Cas9-mediated disruption of Stat6 in IL-4 treated AML cells, we found that the CD47 upregulation was Stat6 dependent, recognizing a previously unknown link between IL-4 signalling and CD47 upregulation. Consistent with this finding, combined antibody blockade of CD47 on leukemia cells and IL-4 stimulation of macrophages resulted in enhanced phagocytosis of AML cells.Taken together, this study shows that IL-4 has anti-leukemic activity by activating macrophages, findings that might translate into new immunotherapeutic opportunities in AML and possibly other malignancies. DisclosuresNo relevant conflicts of interest to declare.

DNA damage induction in bone marrow cells of mice after farnesenes and 2,5-dimethylpyrazine sniffing

Background. Pheromones are an important regulatory link of synecological contacts in numerous animal species. Chemo-signaling participates in establishing of population social structure, it regulates different types of behavior, changes hormonal state and maturation rate, etc. It also can affect the genetic material expression and integrity.&#x0D; Material and methods. Groups of adult males of CBA/Lac/Sto/Rap strain were exposed to volatile chemosignals (mixture of - and -farnesenes or 2,5-dime thylpyrazine) for 2 or 24 hours. Bone marrow cells were prepared for single cell gel electrophoresis (comet assay test). Content of DNA in comet cells were analyzed. In case of 24 hours exposure bone marrow cells were fixed also for ana-telophase analysis.&#x0D; Results. It is shown that exposures with farnesenes or 2,5-DMP both damage genetic material of bone marrow cells. It also followed by induction of mitotic aberration frequency. Simultaneous exposure with all chemosignals does not increase damaging effect.&#x0D; Conclusion. Chemosignals which serve as stress-pheromones in mice decrease also the integrity of genetic material in bone marrow cells of recipients. It could be a mechanism of pheromonal impact on density and space-genetic structure of mouse populations.

Toxicological evaluation of Microbacterium foliorum SYG27B-MF

Microbacterium foliorum is a naturally occurring bacteria in cruciferous vegetables and ripened cheese. The safety of M. foliorum SYG27B-MF has been assessed in both acute and subchronic studies and a battery of mutagenicity and clastogenicity tests. In a single dose acute study, the LD50 of M. foliorum SYG27B-MF was greater than 3 g/kg bw or 5.1 × 1016 colony forming unit (CFU)/kg bw, the highest dose tested. In a 90-day subchronic toxicity study in 80 Sprague-Dawley rats, no animals died and there were no treatment-related abnormalities at doses of 0, 500, 1000, or 2000 mg/kg bw. In a 90-day repeated toxicity test, the no-observed-adverse-effect level (NOAEL) M. foliorum SYG27B-MF was 2000 mg/kg/day or 3.4 × 1016 CFU/kg bw/day, the highest level tested. A mutagenicity study using reverse bacterial mutation tests and a genotoxicity study employing cultured hamster ovarian fibroblasts (CHO-K1) cell showed that M. foliorum SYG27B-MF was not mutagenic or clastogenic in the presence or absence metabolic activation. In an in vivo mouse micronucleus assay, M. foliorum SYG27B-MF did not induce did not induce micronuclei formation in the bone marrow cells of mice, indicating that it is non-clastogenic. The results from these studies support the safety of M. foliorum SYG27B-MF for use as a production organism for human food ingredients.

The genotoxic effects of fruit extract of Crataegus oxyacantha (hawthorn) in mice

ABSTRACT Crataegus oxyacantha L. (Rosaceae) is a medicinal plant with a long history of use in European, Chinese, and American. The majority of pharmacological activities associated with fruit extracts of C. oxyacantha L. are related to cardio-stimulant properties utilized in the treatment of atherosclerosis, hypertension with myocardic insufficiency, angina pectoris, cardiac rhythm alterations, and heart failure. Some other therapeutic uses for renal calculi, dyspnea, as well as a diuretic, sedative, and anxiolytic were also reported. Due to the beneficial potential of C. oxyacantha fruits extract but evidence in vitro of genetic toxicity, the aim of the present study was to examine the genotoxic potential of plant extract in vivo in mice. The extract was administered orally, daily by gavage at doses of 50, 100, and 200 mg/kg body weight for seven days. Data demonstrated that C. oxyacantha extract did not markedly induce DNA damage in leukocytes and bone marrow cells by the comet assay; however, the extract produced a significant rise in micronucleated polychromatic erythrocytes (PCE) at all tested doses in a non-dose dependent manner as evidenced by the micronucleus test. The PCE/normochromatic erythrocytes (NCE) ratio indicated no significant cytotoxicity. Under our experimental conditions, C. oxyacantha fruits extract exhibited weak clastogenic and/or aneugenic effects in bone marrow cells of male mice, confirming our previous in vitro findings that this plant extract induced genotoxicity suggesting that prolonged or high dose use needs to be undertaken with caution.

Deregulated iron metabolism in bone marrow from adenine-induced mouse model of chronic kidney disease.

Although the primary cause of anemia in chronic kidney disease (CKD) is lack of sufficient erythropoietin (EPO), other factors may be involved, including the deregulation of iron metabolism. To clarify the mechanism of deranged erythropoiesis in CKD, we evaluated bone marrow (BM) cells in adenine-induced CKD mice. They showed even higher EPO expression in the kidney. Hepatic hepcidin mRNA and plasma hepcidin and ferritin levels were increased. Flow cytometry revealed a decrease in the number of cells expressing transferrin receptor (TfR), or late erythroid progenitors in BM; these cells correspond to proerythroblasts, and basophilic and polychromatic erythroblasts. In CKD mice, levels of erythroferrone mRNA in BM and splenic cells were significantly decreased, and MafB protein levels in BM cells were significantly increased. These results suggest that, in BM, the decrease in TfR, which may be associated with increased MafB levels, and the decrease in erythroferrone increase hepatic hepcidin expression, which may perturb iron recycling and erythropoiesis.

Establishment of Trace Amount Bone Marrow Hematopoietic Stem Cells of Adult Mouse Transplantation System

To establish a transplantation system for a trace amount of adult mouse bone marrow hematopoietic stem cells (HSCs). The HSCs from bone marrow cells of GFP adult mouse were seperated by using marker combination ESLAM (CD45+CD201+CD150+CD48-) The goal cell population acquired by flow cytometry sorting were injected into lethally irradiated recipient mouse plus protective cells. And the peripheral blood cells from recipient mice tail veins at different time points were acquired to observe the reconstruction level and lineage differentiation. Then the receptor mouse was killed and the cells from thymus, spleen, marrow, and peripheral blood were acquired to measure tissue chimerism. The results of peripheral blood of recipient mice after transplantation showed that the reconstructive efficiency was 100% after 4th week. The proportion of GFP+ cells at different time points in various lines (myeloid, erythroid, B, T and megakaryocytes) were different and the change tendency of reconstruction were different. From different tissues (peripheral blood, bone marrow, spleen, thymus) after 6 months, the intramedullary spleen and thymus chimerism can be seen and showed a high proportion of GFP+cells in the peripheral blood erythroid and megakaryocytes and there were no GFP+cells in the myeloid and lymphatic cells, but the myeloid cells in the bone marrow cells of the recipient mice contained a higher proportion of GFP+cells. The B cells in the spleen had a higher proportion of GFP+ cells, and the T cells in the thymus contained a higher proportion of GFP+ cells. The ESLAM combination can be highly efficient one for the enrichment of HSCs with long-term reconstructive ability, and the results of peripheral blood 6 months after the transplantation suggests that it may have the inclination to erythroid and megakaryocyte differentiation.

Antimutagenic efficacy of purified anthocyanin From the in vitro culture of teak leaves on b Marrow cells of mice

Antimutagenic efficacy of purified anthocyanin From the in vitro culture of teak leaves on b Marrow cells of mice

Predator Odor Destabilizes the Cell Genome of the Mouse Bone Marrow

The effect of urine chemosignals of domestic male cats (Felis catus L.) on the stability of the bone marrow cell genome and corticosterone level in blood plasma of recipient mice (Mus musculus L.) is studied. It is demonstrated that a two-hour sniffing of volatile chemosignals leads to an increase in the frequency of bone marrow cells with damaged DNA, while 24 h exposure to chemosignals leads to an increase of chromosomal aberrations frequency in these cells. Thus, the effect of genomic instability induction in bone marrow cells of house mice by predator’s chemosignals is for the first time demonstrated in the work. At the same time, no increase in the corticosterone level is detected in the blood plasma either 30 or 60 min after the beginning of the exposure to the chemosignals of cat urine (CU). The physiological factors contributing to DNA damages accumulation in bone marrow cells, as well as the long-term consequences of the effect of these chemosignals on the genome stability, are discussed.

Effect of Carbafuran Pesticide on Mitotic Chromosomes of Mus musculus

The Genotoxic effect of the carbafuran pesticide was evaluated in the metaphasic chromosomes of bone marrow cells of female Swiss albino mice, Mus musculus . Six week old mice were put into different groups(six mice per group) and treatment were done according to experimental plan (CLD15 i.e. lower dose =30ppm or 0.03mg/ml and CHD30 i.e. higher dose =300ppm or 0.3mg/ml).The test material was administered orally for 15 days in CLD15 and for 30 days in CHD30 test groups of female mice. The result revealed that the frequency of chromosomal abnormalities i.e. structural and numerical changes were11.5% in control, 74.0%in CLD15 and 99.0%in CHD30, respectively in bone marrow cells of female mice.Int. J. Appl. Sci. Biotechnol. Vol 6(2): 169-173

Inducible nitric oxide synthase during the late phase of sepsis is associated with hypothermia and immune cell migration

Hypothermia is a significant sign of sepsis, which is associated with poor prognosis, but few mechanisms underlying the regulation of hypothermia are known. Inducible nitric oxide synthase (iNOS) is a key inflammatory mediator of sepsis. However, the therapeutic benefit of iNOS inhibition in sepsis is still controversial, and requires elucidation in an accurate model system. In this study, wild-type (WT) mice showed temperature drops in a biphasic manner at the early and late phase of sepsis, and all mice died within 48 h of sepsis. In contrast, iNOS-knockout (KO) mice never showed the second temperature drop and exhibited improved mortality. Plasma nitric oxide (NO) levels of WT mice increased in the late phase of sepsis and correlated to hypothermia. The results indicate that iNOS-derived NO during the late phase of sepsis caused vasodilation-induced hypothermia and a lethal hypodynamic state. The expression of the iNOS mRNA was high in the lung of WT mice with sepsis, which reflects the pathology of acute respiratory distress syndrome (ARDS). We obtained the results in a modified keyhole-type cecal ligation and puncture model of septic shock induced by minimally invasive surgery. In this accurate and reproducible model system, we transplanted the bone marrow cells of GFP transgenic mice into WT and iNOS-KO mice, and evaluated the role of increased pulmonary iNOS expression in cell migration during the late phase of sepsis. We also investigated the quantity and type of bone marrow-derived cells (BMDCs) in the lung. The number of BMDCs in the lung of iNOS-KO mice was less than that in the lung of WT mice. The major BMDCs populations were CD11b-positive, iNOS-negative cells in WT mice, and Gr-1-positive cells in iNOS-KO mice that expressed iNOS. These results suggest that sustained hypothermia may be a beneficial guide for future iNOS-targeted therapy of sepsis, and that iNOS modulated the migratory efficiency and cell type of BMDCs in septic ARDS.

Mequindox Induced Genotoxicity and Carcinogenicity in Mice.

Mequindox (MEQ), acting as an inhibitor of deoxyribonucleic acid (DNA) synthesis, is a synthetic heterocyclic N-oxides. To investigate the potential carcinogenicity of MEQ, four groups of Kun-Ming (KM) mice (50 mice/sex/group) were fed with diets containing MEQ (0, 25, 55, and 110 mg/kg) for one and a half years. The result showed adverse effects on body weights, feed consumption, hematology, serum chemistry, organ weights, relative organ weights, and incidence of tumors during most of the study period. Treatment-related changes in hematology, serum chemistry, relative weights and histopathological examinations revealed that the hematological system, liver, kidneys, and adrenal glands, as well as the developmental and reproductive system, were the main targets after MEQ administration. Additionally, MEQ significantly increased the frequency of micronucleated normochromatic erythrocytes in bone marrow cells of mice. Furthermore, MEQ increased the incidence of tumors, including mammary fibroadenoma, breast cancer, corticosuprarenaloma, haemangiomas, hepatocarcinoma, and pulmonary adenoma. Interestingly, the higher incidence of tumors was noted in M25 mg/kg group, the lowest dietary concentration tested, which was equivalent to approximately 2.25 and 1.72 mg/kg b.w./day in females and males, respectively. It was assumed that the lower toxicity might be a reason for its higher tumor incidence in M25 mg/kg group. This finding suggests a potential relationships among the dose, general toxicity and carcinogenicity in vivo, and further study is required to reveal this relationship. In conclusion, the present study demonstrates that MEQ is a genotoxic carcinogen in KM mice.