P5746Human induced pluripotent stem cell-derived mesenchymal stem cell therapy effectively reduced brain infarct volume and preserved neurological function in rat after acute intracranial hemorrhage

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
Y C Li ◽  
F Y Lee ◽  
S Chua ◽  
H K Yip
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Neurological Function ◽  
Opposite Pattern ◽  
Group 4 ◽  
Tnf Α ◽  
Group 3 ◽  
Group 2 ◽  
Induced Pluripotent

Abstract Intracerebral hemorrhage (ICH) causes 10%-20% of all strokes and results in higher morbidity compared to other subtypes of cerebral stroke. Although early surgical intervention can clear the expanding hematoma, clinical outcomes following ICH have not significantly improved over the decades. Since ICH elicits neuroinflammation to exacerbate brain edema, damage the blood-brain barrier (BBB), lead to secondary neuronal injury, anti-inflammation may be a critical therapeutic strategy. Mesenchymal stem cell (MSC) therapy processes anti-inflammatory, immunomodulatory and tissue regenerative properties, suggesting that MSC therapy could be an effective therapy for ICH. Therefore, this study tested the hypothesis that human induced pluripotent stem cell-derived mesenchymal stem cell (iPSC-MSC) therapy could effectively reduce brain-infract volume (BIV) and improve neurological function in rat after acute ICH induced by a weight-drop device. Adult-male SD rats (n=40) were equally divided into group 1 (sham-operated control), group 2 (ICH), group 3 (ICH + hyaluronic acid (HA)/intracranial injection/3h after ICH), group 4 [ICH + HA + iPSC-MSC (1.2x106 cells/intracranial injection/3h after ICH)] and euthanized by day 28 after ICH procedure. In vitro study showed that hemorrhagic-brain tissue augmented protein expressions of inflammation (HMGB1/MyD88/TLR-4/TLR-2/NF-κB/TNF-α/iNOS/IL-1β) in cultured neurons that were significantly inhibited by iPSC-MSC treatment (all p<0.001). By days 7/14 after ICH procedure, circulating inflammatory levels of TNF-α/IL-6/MPO expressed were lowest in group 1, highest in group 2 and significantly lower in group 4 than in group 3 (all p<0.0001). By day 14 after ICH procedure, neurological function and BIV expressed an opposite pattern, whereas protein expressions of inflammation (HMGB1/MyD88/TLR-4/TLR-2/NF-κB/I-kB/TNF-α/iNOS/IL-1β/MMP-9), oxidative stress (NOX-1/NOX-2/oxidized protein) and apoptosis (mitochondrial-Bax/cleaved-caspase-2/PARP) in brain exhibited an identical pattern to circulating inflammation among the four groups (all p<0.001). Microscopy demonstrated that the number of vascular remodeling/GFAP+/53BP1+/γ-H2AX+ cells displayed an identical pattern of inflammation, whereas the NeuN+ cells displayed an opposite pattern of inflammation among the four groups (all p<0.001). In conclusion, iPSC-MSC therapy markedly reduced BIV and preserved neurological function mainly by inhibiting inflammatory/oxidative-stress generation. Acknowledgement/Funding Kaohsiung Chang Gung Memorial Hospital, Taiwan Society of Stem Cell Research

scholarly journals Additional benefit of induced pluripotent stem cell-derived mesenchymal stem cell therapy on sepsis syndrome-associated acute kidney injury in rat treated with antibiotic

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chih-Chao Yang ◽  
Pei‐Hsun Sung ◽  
Chih-Hung Chen ◽  
John Y. Chiang ◽  
Pei-Lin Shao ◽  
...  
Keyword(s):  
Stem Cell ◽  
Kidney Injury ◽  
Induced Pluripotent Stem Cell ◽  
Inflammatory Cells ◽  
Sepsis Syndrome ◽  
Group 4 ◽  
Group 6 ◽  
Group 2 ◽  
Induced Pluripotent ◽  
Group 1

Abstract Background This study tested whether human induced-pluripotent stem-cell-derived mesenchymal-stem-cells (iPS-MSCs) would offer an additional benefit to the rodent with acute kidney injury (AKI) (ischemia for 1 h followed by reperfusion for 120 h) associated sepsis syndrome (SS) (by cecal-ligation-puncture immediately after AKI-induction) undergoing ciprofloxacin therapy. Results Male-adult SD rats (n = 80) were categorized into group 1 (sham-operated-control, n = 10), group 2 (AKI + SS, n = 24), group 3 (AKI + SS + ciprofloxacin/3 mg/kg, orally for 120 h, n = 12), group 4 (AKI + SS + iPS-MSCs/1.2 × 106/intravenously administered by 3 h after AKI, n = 12), group 5 (AKI + SS + iPS-MSCs/1.2 × 106/intravenously administered by 18 h after AKI, n = 12), group 6 (AKI + SS + iPS-MSCs/1.2 × 106/intravenously administered by 3 h after AKI induction + ciprofloxacin, n = 10] and euthanized by 120 h. The result showed that the mortality was significantly higher in group 2 than in other groups (all p < 0.01). The creatinine level was highest in group 2, lowest in group 1, significantly lower in group 6 than in groups 3, 4 and 5, (all p < 0.0001), but it showed no difference among the latter 3 groups. Flow cytometric analysis showed that the circulatory inflammatory cells (Ly6G/CD11b/c), early (AN-V+/PI−)/late (AN-V+/PI+) apoptosis, and circulatory/splenic immune cells (CD3+/CD4+, CD3+/CD8a+) were highest in group 2, lowest in group 1, significantly lower in group 6 than in groups 3/4/5 and significantly lower in group 4 than in groups 3/5 (all p < 0.0001), but they showed no difference between groups 3/5. Protein expressions of oxidative-stress (NOX-1/NOX2/oxidized protein), apoptotic (cleaved-caspase3/cleaved-PARP/mitochondrial-Bax), fibrotic (TGF-ß/Smad3), inflammatory (MMP-9/IL-6/TNF-α) and autophagic (Atg5/Beclin) biomarkers in kidney exhibited an identical pattern of circulatory inflammatory cells (all p < 0.0001). Conclusion Combined iPS-MSCs-ciprofloxacin therapy was superior to either one alone for protecting AKI complicated by SS.

scholarly journals Acute Phase Effect of Memantine Hydrochloride in Experimental Peripheral Nerve Injury

2019 ◽  
Vol 08 (02) ◽  
pp. 113-118
Author(s):  
Hakan AK ◽  
Iskender Samet Daltaban ◽  
Sevilay Vural
Keyword(s):  
Peripheral Nerve ◽  
Nerve Injury ◽  
Peripheral Nerve Injury ◽  
Group 4 ◽  
Tnf Α ◽  
Group 3 ◽  
The Mean ◽  
Cox 2 ◽  
Group 2 ◽  
Group 1

Abstract Aim In this experimental study, we aimed to investigate possible healing effects of memantine hydrochloride, an N-methyl-d-aspartate (NMDA) antagonist, with clinical, biochemical, and histopathologic methods on acute peripheral nerve injury (PNI). Material and Method Forty-eight adult Wistar albino rats were divided into four groups (n = 12). The groups were arranged as sham-operated group (group 1), acute compression model group (group 2), trauma + low-dose memantine group (group 3), and trauma + high-dose memantine group (group 4). Memantine was administered intraperitoneally for 7 days. Subjects were sacrificed after the measurement of the sciatic nerve function index (SNFI) on the eighth day. Cyclooxygenase 2 (COX-2) and tumor necrosis factor-α (TNF-α) levels were measured in nerve tissues. Histopathologic evaluation was performed by electron microscopy. Results The mean sciatic function index (SFI) scores of groups 1 to 4 were +3.27 (standard deviation [SD] ±4.66),–18.2 (SD = ±11.7),–8.5 (SD = ±7.5), and–2.5 (SD = ±9), respectively. The mean COX-2 values were 0.98 ng/mL (SD = ±0.51), 1.89 ng/mL (SD = ±0.22), 1.39 ng/mL (SD = ±0.36), and 1.35 ng/mL (SD = ±0.59), respectively. TNF-α values were 0.09 pg/mL (SD = ±0.23), 1 pg/mL (SD = ±0.96), 0.46 pg/mL (SD = ±0.55), and 0.48 pg/mL (SD = ±0.78), respectively. Group 1 showed normal histologic findings. Group 2 showed marked edema particularly in large-diameter myelins. Myelin configurations were detected in large myelinated axons in group 3. The number of mast cells in endoneurium was high in group 4. Conclusion The efficacy of memantine in the acute phase of PNI appears to be significant according to the SNFI and biochemical tests. However, histologic findings suggest that high doses of memantine have a negative effect on PNI.

scholarly journals Extracorporeal Shock Wave Therapy Protected the Functional and Architectural Integrity of Rodent Urinary Bladder against Ketamine-Induced Damage

Biomedicines ◽  
2021 ◽  
Vol 9 (10) ◽  
pp. 1391
Author(s):  
Yen-Ta Chen ◽  
Kuan-Hui Huang ◽  
John Y. Chiang ◽  
Pei-Hsun Sung ◽  
Chi-Ruei Huang ◽  
...  
Keyword(s):  
Urinary Bladder ◽  
Cyclophilin D ◽  
Group 4 ◽  
Extracorporeal Shock Wave ◽  
Tnf Α ◽  
Bladder Contraction ◽  
Group 3 ◽  
Group 2 ◽  
Cell Stress Response ◽  
Group 1

This study tested the hypothesis that extracorporeal-shock-wave (ECSW) protected the functional and anatomical integrity of rat urinary-bladder against ketamine-induced damage. In in vitro study, the rat bladder smooth muscle cells (RBdSMCs) were categorized into G1 (sham-control), G2 (RBdSMCs + menadione), G3 (RBdSMCs + ECSW) and G4 (RBdSMCs + menadione + ECSW). The results showed protein expressions of oxidative-stress/mitochondrial-damaged biomarkers (NOX-1/NOX-2/oxidized protein/cytosolic-cytochrome-C/cyclophilin-D), inflammatory markers (MyD88/TRAF6/p-IKB-α/NF-κB/TNF-α/IL-6/IL-1ß/MMP-9/iNOS), and cell-stress response signalings (ASK1/p-MKK4/p-MKK7/ERK1/2//p-JNK/p-p38/p-53) were significantly increased in G2 than in G1 and G3, and those were significantly reversed in G4 (all p < 0.0001). Adult-male SD rats (n = 24) were equally categorized into group 1 (sham-control), group 2 (ketamine/30 mg/kg/daily i.p. injection for four weeks), group 3 [ketamine/30 mg/kg + ECSW/optimal energy (0.12 mJ/mm2/120 impulses/at 3 h and days 3/7/14/21/28 after ketamine administration)] and group 4 [(ketamine/30 mg/kg + ECSW/higher energy (0.16 mJ/mm2/120 impulses)] and animals were euthanized by day 42. The results showed the urine levels of pro-inflammatory cytokines (TNF-α/IL-6) were lowest in group 1, highest in group 2 and significantly higher in group 3 than in group 4 at days 1/7/14/28 (all p < 0.0001). The duration of urinary bladder contraction was lowest in group 2, highest in group 1 and significantly higher in group 4 than in group 3, whereas the maximal pressure of urinary bladder exhibited an opposite pattern of bladder contraction among the groups (all p < 0.0001). The histopathological findings of fibrosis/inflammation/keratinization and protein expressions of oxidative-stress/mitochondrial-damaged biomarkers (NOX-1/NOX-2/oxidized protein/cytosolic-cytochrome-C/cyclophilin-D), and inflammatory (TLR-2/TLR-4/MyD88/TRAF6/p-IKB-α/NF-κB/TNF-α/IL-1ß/MMP-9/iNOS) and cell-stress response (ASK1/p-MKK4/p-MKK7/ERK1/2//p-JNK/p-p38) signalings and apoptotic/fibrotic biomarkers (cleaved-caspas3/cleaved-PARB/Smad3/TFG-ß) exhibited an identical pattern of urine proinflammatory cytokine among the groups (all p < 0.0001). ECSW effectively attenuated ketamine-induced bladder damage and dysfunction.

Impact of Peripheral Blood Stem Cell Recruitment on Outcome of Single or Double Autologous Hematopoietic stem Cell Transplantation for Multiple Myeloma.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 5221-5221
Author(s):  
Mauricette Michallet ◽  
Quoc-Hung Le ◽  
Anne-Sophie Michallet ◽  
Anne Thiebaut ◽  
Emmanuelle Tavernier ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Multivariate Analysis ◽  
Stem Cell ◽  
Light Chains ◽  
Hematopoietic Stem ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Abstract Multiple myeloma remains one of the best indication for intensive chemotherapy followed by autologous hematopoietic stem cell transplantation (autoT). Intensive therapy followed by autologous transplantation is superior to conventional chemotherapy and it was demonstrated that two autoT were superior to one except for patients in very good partial response or in complete response after the first autotransplant. Peripheral blood stem cells (PBSC) can be used as well as bone marrow as HSC source with the same efficacy but very few data have been reported regarding PBSC recruitment. The main goal of our work was to study the impact on overall and event-free survival (OS and EFS) of PBSC recruitment using either growth factors (GF) alone (steady state) or chemotherapy followed by GF. Secondly, we performed a multivariate analysis studying influence on OS and EFS of sex, age, lines of therapy, pretransplant status, TBI, PBSC recruitment and number of autoT. We have analyzed 193 PBSC autoT (1 autoT=160, 2 autoT=33) performed for 160 MM patients [81 males and 71 females, mean age: 55 years (39–71)]. At diagnosis, 88 patients presented a MM Ig G (70k and 18l), 28 Ig A (16k and 12l), 3 Ig D (1k and 2l), 21 light chains k and 13 light chains l, 3 non secreting and 4 with plasmocyte leukemias. According to Durie and Salmon classification 75% of patients were in stage III, 15% in stage II and 10% in progressive I. Before transplantation, patients have received 1 line of poly-chemotherapy (n=141), 2 lines (n=15) or 3 lines (n=4) and 154 were evaluated for the response with 11 complete remission, 113 partial remission and 30 stable or evolutive disease just before transplant. As HSC (n=189), patients received PBSC which were recruited by GF alone (n=105) or cyclophosphamide+GF (n= 84). Conditioning (n=189),consisted in melphalan and TBI (n=51), melphalan alone (n=132), melphalan associated to cyclophosphamide or busulfan (n=6). We divided the population into 4 groups : group 1 who received one autoT of PBSC recruited by GF (n=76), group 2 one autoT of PBSC recruited by chemotherapy+GF (n=50), group 3 two autoT of PBSC recruited by GF (n=16) and group 4 two autoT of PBSC recruited by chemotherapy+GF (n=17). The median follow-up (FU) of the 4 groups were different with shorter FU (group 3: 9.9 months, group 4: 13 months) for patients who received tandem autoT because of the recent character of this strategy as compared to a long term follow-up for patients who received only one transplant (group 1: 35months, group 2: 55.3 months). Probabilities of OS and EFS at 2 years were 76% (95%CI 67–87) and 60% (95%CI 49.5–73) for group 1, 77% (95%CI 65–90.5) and 70% (95%CI 57.5–85) for group 2, 87.5% (95%CI 73–100) and 72.9% (95%CI 49–100) for group 3, 100% and 66.7% (95%CI 36–100) for group 4. The difference was not significant because of follow-up differences between the 4 groups and small number of patients in groups 3 and 4. In addition, multivariate analysis did not show any significant influence of the different studied parameters on OS and EFS. Nevertheless, because of these interesting preliminary results, a longer follow-up is warranted for definitive conclusions.

Leukemic and Hematopoietic Stem Cells Compete for the Same Functional Marrow Niche in a MLL-AF9 Murine Leukemia Model

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4897-4897
Author(s):  
Ronan G. Desmond ◽  
Taha Bat ◽  
Olena Kamenyeva ◽  
Benjamin Mizukawa ◽  
James C. Mulloy ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Murine Model ◽  
Leukemic Cells ◽  
Hematopoietic Stem ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Abstract Abstract 4897 Much is known regarding the location, cellular composition, signaling pathways, and functional role of the normal hematopoietic stem cell (HSC) niche in the bone marrow microenvironment. Microenvironmental cells including osteoblasts, other specialized mesenchymal cells, and vascular endothelial cells exert control over HSC self-renewal, differentiation, and engraftment. Niche occupancy appears to be competitive and limiting in terms of controlling the number of HSCs per organism. Leukemia stem cells (LSCs), through their inherent properties of quiescence and resistance to chemotherapeutic agents, are thought to be one of the principal mechanisms underlying disease relapse in patients. Much less is known regarding the interaction of LSCs and the marrow microenvironment. It is not clear whether LSCs localize to the same niches as HSCs, compete with HSCs for niche occupancy, or share dependence on niche signals, and whether those signals affect tumor responses to chemotherapy. Using a human pre-B ALL xenograft mouse model, Colmone et al (Science 2008) recently showed that leukemic cells may alter the normal microenvironment, resulting in initial homing of transplanted normal HSPCs in distinct atypical niches. Shiozawa et al (JCI 2011) showed that metastatic prostate cancer cells, a tumor type known to target bone, impeded HSC engraftment in a murine model, suggesting competition for the same niche. To investigate the relationship between HSC and LSC niche localization and functional occupancy, we used murine progenitor cells transduced with an MLL-AF9 vector expressing GFP in a murine syngeneic competitive transplantation model. MLL-AF9 cells are highly enriched for LSCs, particularly the c-kit+ compartment (Somervaille Cancer Cell 2006). We found that between approximately 21% and 24% of cells were c-kit+ by FACS in 2 separate experiments. In our model, mice transplanted with unsorted MLL-AF9 cells (1×107) died of AML with a latency of 11–14 days. We cotransplanted a fixed number of MLL-AF9-GFP cells (1×106) with increasing numbers of normal mouse whole bone marrow (WBM) cells, derived from dsRed transgenic mice to facilitate distinction from the GFP+ MLL-AF9 cells, into mice irradiated with 1000 rads: 1×105 [group 1], 1×106 [group 2], 1×107 [group 3], 5×107 [group 4]. Control groups received 1×105 and 1×106 normal WBM cells only. Survival was monitored daily. The control group receiving 1×105 cells only all died with median time to death of 16.5 days from lack of count recovery, those receiving 1×106 cells are still alive 35 days after transplant, indicating that 1×106 cells is adequate to rescue from irradiation. Mice were bled weekly until death and samples were analyzed by flow cytometry. Complete blood counts, blood smears, and splenic sections were obtained from these mice. As expected, there were no circulating blasts detected 7 days post transplant and all mice were healthy. However, 14 days after transplant the percentages of GFP+ leukemic cells detected in the blood were inversely proportional to the number of normal dsRed WBM cells transplanted (group 1 vs. group 2 vs. group 3 vs. group 4 mean percentage of GFP+ cells, 83.97 v 66.53 v 18.73 v 9.275 p< 0.0001). At day 15, mice from group 1, but not from groups 2 to 4, became moribund and were sacrificed. Spleens in this group were heavier than in those mice transplanted with 1×105 normal WBM cells alone and 2 out of 3 showed leucocytosis compared to leucopenia in all mice in the group transplanted with normal cells alone. When mice in the other groups had blood samples taken for analysis while moribund, GFP+ cells were greater than 80% suggesting that mice in group 1 died from complications relating to leukemic infiltration. Confocal microscopy confirmed the colocalization of normal HSPCs and MLL-AF9-GFP LSCs in the niche. Most interestingly, survival was proportional to the numbers of normal WBM cells transplanted, with a continuous delay in leukemic death proportional to the number of normal WBM cells cotransplanted with the same dose of MLL-AF9 cells (Figure 1). Hence, this murine model of leukemia suggests that normal and leukemic cells compete for the same functional niche, that manipulation of the niche could impact on response to anti-leukemic therapies, and that cell dose in the context of stem cell transplantation for leukemia may have an impact on outcome via niche competition. Figure 1 Figure 1. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Changes in the immunomorphological indicators of the placenta in women with exacerbation of cytomegalovirus infection in the second trimester of pregnancy complicated by chronic placental insufficiency

2021 ◽  
pp. 80-85
Author(s):  
I. N. Gorikov ◽  
I. A. Andrievskaya
Keyword(s):  
Cytomegalovirus Infection ◽  
Placental Insufficiency ◽  
Second Trimester ◽  
Chorionic Villi ◽  
Group 4 ◽  
Terminal Villi ◽  
Tnf Α ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Aim. To assess the change of immunomorphological parameters in the placenta in women with exacerbation of cytomegalovirus infection (CMVI) in the second trimester of pregnancy, complicated by chronic placental insufficiency.Materials and methods. The concentration of TNF-α in the homogenate of 120 placentas and the histometric parameters of chorionic villi were determined in patients who underwent latent CMVI and exacerbation of CMVI in the second trimester of gestation. Group 1 included 30 placentas from seronegative women with a physiological course of pregnancy, group 2 included 30 placentas from patients with latent CMVI and chronic compensated placental insufficiency, group 3 – 30 placentas from women with exacerbation of CMVI and chronic compensated placental insufficiency; and group 4 – 30 placentas from pregnant women with exacerbation of CMVI and chronic subcompensated placental insufficiency.Results. In the 1st group in the placenta homogenate the concentration of TNF-α was 16.8±1.86 pg/mL; the number of villi with a diameter of 30-50 microns was 25.4±2.08%, with a diameter of 60-90 microns – 64.4±2.43% and with a diameter of more than 90 microns – 10.2±0.88%; the number of terminal villi with 1-3 capillaries was 27.0±2.29%, with 4- 6 capillaries – 42.1±2.02%, with 7-10 capillaries – 23.9±1.58% and villi with more than 10 capillaries – 7.0±0.79%. In group 2, the concentration of TNF-α in the placenta homogenate was amounted to 22.1±2.06 pg/mL (p>0.05); among the villi, anatomical forms with a diameter of 30-50 μm (p<0.01) were found 1.41 times more often, and villi with a diameter of 60-90 μm (p<0.01) were 1.19 times less common; the number of villi with 4-6 capillaries decreased by 1.21 times (p<0.05) and the number of villi with 7-10 capillaries increased by 1.43 times (p<0.001). In the placentas of group 3, compared with group 2 in the homogenate, there was an increase in the concentration of TNF-α to 60.2±3.47 pg/mL (p<0.001) against the background of a decrease in the concentration of villi with a diameter of 30-50 μm to 26.4±2,61% (p<0.05), villi with 7-10 capillaries up to 20.7±1.53% (p<0.001) and an increase in the number of villi with 1-3 capillaries up to 34.8±3.05% (p<0.05). In the placental homogenate of group 4, compared with group 3, the concentration of TNF-α (p<0.05) increased 1.31 times, the number of villi with a diameter of 60-90 μm increased to 70.2±1.59%, (p<0,01) and villi with 1- 3 capillaries to 46.8±3.76% (p<0.05) with a decrease in the number of villi with a diameter of 30-50 μm to 18.9±1.69% (p<0,05), with 7-10 capillaries up to 13.3±1.36% (p<0.001) and with 10 or more capillaries – up to 3.9±0.43% (p<0.01).Conclusion. In women with exacerbation of CMVI in the second trimester of pregnancy and the development of chronic subcompensated placental insufficiency, inhibition of the growth and angiogenesis of terminal villi is observed against the background of the maximum concentration of TNF-α in the medium.

Characterisation of Leukaemic Stem Cell Populations in Childhood AML Indicate Distinct Disease Subtypes with Diverse Clinical Features

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1500-1500
Author(s):  
Victoria J Weston ◽  
Tracey A Perry ◽  
Katie Brown ◽  
Shaun R Wilson ◽  
Pamela R Kearns
Keyword(s):  
Stem Cell ◽  
Cd38 Expression ◽  
Group 4 ◽  
Childhood Aml ◽  
Group 3 ◽  
The Mean ◽  
Group 2 ◽  
Stem Cell Populations ◽  
Group 1

Abstract Abstract 1500 Five year survival rates for childhood AML in children are currently 55–65%. AML is an extremely heterogeneous disease, and while prognostic significance of some karyotypic abnormalities has become evident, the biology of the disease remains largely unknown. Full characterisation of leukaemia initiating cells which may be responsible for relapse has not yet been undertaken. We investigated the leukaemic stem cell populations from 10 childhood primary AML samples by comparing expression of CD34, CD38 and CD45RA, a marker of a committed granulocyte-macrophage progenitor (GMP)-like population frequent in adult AML, in vitro daunorubicin sensitivity and engraftment in immuno-compromised NOD/Shi-scid/IL-2Rgnull (NOG) mice. Consequently, we were able to classify AML samples into 4 subgroups. These comprised Group 1, CD34+CD38- AML (n=1); Group 2, CD34-CD38+CD45RA- (<10% CD34+ blasts) AML (n=4); Group 3, CD34+CD38+CD45RA- (>10% CD34+ blasts) AML (n=4); and Group 4, CD34-CD38+CD45RA+ (<10% CD34+ and >10% CD45RA+ blasts) (n=1). There was no apparent enrichment for high risk prognostic karyotypes in any of the groups. The Group 1 AML presented at 3y with t(16;21); In Group 2 AMLs, the mean presentation age was 11y, 2 carried good prognostic t(8;21), while 1 had MLL involvement and 1 had FLT3-ITD with chromosome 13 isodisomy, both higher risk indicators; The Group 3 AMLs presented with a mean age of 11.9y and 2 carried good prognostic inv(16) whereas 2 had FLT3-ITD one with additional chromosome 13 isodisomy, t(5;11) and TP53 loss. Finally, the Group 4 AML presented at 1.5y with a normal karyotype. When we compared the 2 most frequent subgroups, Group 2 had a much shorter mean EFS of 122d (n=2) compared with a 275d (n=4) for Group 3 (the mean follow up was 282d and 1013.5d, respectively). We next sorted four cell subpopulations based on CD34 and CD38 expression (CD34+CD38-, CD34+CD38+, CD34-CD38- and CD34-CD38+ blasts) and compared in vitro sensitivity to daunorubicin. In Group 1, CD34- and CD34+ cells were equally sensitive at nanomolar IC50 doses. In 2 of the Group 2 AMLs,CD34-CD38- cells were the most resistant to daunorubicin at micromolar IC50 doses (2.5-10mM) whereas the CD34-CD38+ cells (also the dominant subpopulation in this group) were the most sensitive cells exhibiting nanomolar IC50 doses (750–800nM). In contrast, the Group 3 AMLs were overall more sensitive to daunorubicin exhibiting lower nanomolar IC50 doses. Again in this group, the CD34-CD38- cells were typically the most resistant (this time being the dominant subpopulation) whereas the CD34+CD38+ were the most sensitive cells. Finally, in the Group 4 AML, while CD45RA+ cells rapidly underwent spontaneous apoptosis, CD45RA- cells exhibited extreme resistance to daunorubicin (IC50 >10mM) and CD38 expression had no impact on sensitivity. The reduced sensitivity of Group 2 AMLs to daunorubicin compared with Group 3 could, therefore, be an underlying factor contributing to shorter EFS. Finally, we initiated comparison of the stem cell qualities of the different subpopulations from representative samples from each of the two major subgroups, first by assessment of differentiation potential in vitro, and second by engraftment in vivo using NOG mice. In on-going experiments, the time to leukaemia will be compared between mice injected with unsorted and sorted cells and, at terminal cull, cells harvested from organs will be characterised by karyotype and immunophenotype and tested for clonogenic potential via subsequent serial transplantations. Peripheral blood sampling currently suggests higher human CD45+ engraftment in mice injected with sorted versus unsorted cells, and these are CD34+CD33+CD3- recapitulating the AML phenotype. We anticipate that particular subpopulations will be enriched for AML stem cells with the ability to repopulate the leukaemia. Overall, we have shown that childhood AML is diverse with respect to stem cell characteristics. AMLs with low CD34 (Groups 2 and 4) exhibit the greatest overall resistance to daunorubicin as well as shorter EFS. Furthermore, in the majority of AMLs, CD34-CD38- blasts exhibit the least sensitivity to daunorubicin. Novel therapies which can target these resistant subpopulations with leukaemia initiating activity could significantly improve the treatment responses in this clinically challenging disease. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Effect of cogon grass root ethanol extract on fatty acid binding protein 4 and oxidative stress markers in a sepsis mouse model

F1000Research ◽  
2022 ◽  
Vol 10 ◽  
pp. 1161
Author(s):  
Mirasari Putri ◽  
Bening Mauliddina Rastiarsa ◽  
Raden Aliya T. M. Djajanagara ◽  
Ghaliby Ardhia Ramli ◽  
Neni Anggraeni ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Mouse Model ◽  
Fatty Acid Binding ◽  
Stress Markers ◽  
Group 4 ◽  
Tnf Α ◽  
Grass Root ◽  
Group 3 ◽  
Group 2 ◽  
And Oxidative Stress

Background: Sepsis causes several immunological and metabolic alterations that induce oxidative stress. The modulation of fatty acid-binding protein 4 (FABP4) has been shown to worsen this condition. Extract of cogon grass root (ECGR) contains flavonoids and isoeugenol compounds that exhibit anti-inflammatory and antioxidant properties. This study aimed to assess the effects of ECGR on FABP4 and oxidative stress–related factors in a sepsis mouse model. Methods: Twenty-nine male mice (Mus musculus) of the Deutsche Denken Yoken strain were divided into four groups: group 1, control; group 2, mice treated with 10 μL/kg body weight (BW) lipopolysaccharide (LPS); and groups 3 and 4, mice pre-treated with 90 and 115 mg/kg BW, respectively, and then treated with 10 μL/kg BW LPS for 14 d. Blood, liver, lymph, and cardiac tissue samples were collected and subjected to histological and complete blood examinations. Antioxidant (Glutathione peroxidase 3 (GPx3) and superoxide dismutase), FABP4 levels, and immune system-associated biomarker levels (TNF-α, IL-6 and IL-1β ) were measured. Results: Significant increases in platelet levels (p = 0.03), cardiomyocyte counts (p =0.004), and hepatocyte counts (p = 0.0004) were observed in group 4 compared with those in group 2. Conversely, compared with those in group 2, there were significant decreases in TNF-α expression in group 3 (p = 0.004), white pulp length and width in group 4 (p = 0.001), FABP4 levels in groups 3 and 4 (p = 0.015 and p = 0.012, respectively), lymphocyte counts in group 4 (p = 0.009), and monocyte counts (p = 0.000) and polymorphonuclear cell counts in the livers (p = 0.000) and hearts (p = 0.000) of groups 3 and 4. GPx3 activity was significantly higher in group 3 than in group 1 (p = 0.04). Conclusions: ECGR reduces FABP4 level and modulating oxidative stress markers in sepsis mouse model.

scholarly journals Neurotoxic Assessment of Chronic Abuse of Pregabalin in Wistar Rats

2021 ◽  
pp. 58-66
Author(s):  
Neveen A. Salem ◽  
Amani M. Alsaedi ◽  
Bedor G. Alasmari ◽  
Razan Z. Almarghalani ◽  
Shahad M. Algobe ◽  
...  
Keyword(s):  
Brain Tissue ◽  
Wistar Rats ◽  
Gamma Aminobutyric Acid ◽  
Stress Markers ◽  
Group 4 ◽  
Tnf Α ◽  
Male Wistar Rats ◽  
Group 3 ◽  
Group 2 ◽  
Antioxidant Markers

Pregabalin (Lyrica) is an analog of the gamma-aminobutyric acid neurotransmitter,   approved for the treatment of epilepsy, generalized anxiety disorder, neuropathic pain, and fibromyalgia. The possibility for abuse and/or dependence on pregabalin has risen recently. Pregabalin is controlled in many countries including Saudi Arabia. However, unofficial use of this substance is also on the increase. The purpose of this study is to assess the potential neurotoxic effects associated with overdose prolonged pregabalin supplementation. Forty male Wistar rats were divided into Group (1) normal control received distilled water, Group (2) received pregabalin (150mg/kg), Group (3) received pregabalin (300 mg/kg), and Group (4) received pregabalin (600 mg/kg). pregabalin consumption in different doses resulted in significant dysregulation in neurotransmitter release, upsurge oxidative stress markers via enhancing lipid peroxidation and depleting antioxidant markers. Also, pregabalin doses evoked brain tissue inflammation through elevating TNF-α, IL-1β, and MCP-1, Moreover promoted brain tissue apoptosis by activating caspase -3 and suppressed Bcl2. Pregabalin effects on the aforementioned parameters were dose-dependent. These findings could highlight the potential neurotoxic effect of prolonged abuse of pregabalin supplementation through dysregulating brain neurochemical, inflammatory, oxidant/antioxidant, and apoptotic mediators.

Sign in / Sign up

Export Citation Format

Share Document