PROSPECTIVE SINGLE CENTER ANALYSIS OF OUTCOME STEM CELLS TRANSPLANTS IN PATIENTS WITH CEREBRAL PALSY.

OBJECTIVE: Aim: To evaluate efficacy and safety of autologous bone marrow-derived mononuclear stem cell transplantation intrathecal in children with cerebral palsy. PATIENTS AND METHODS: Materials and Methods: 35 children have levels I-V cerebral palsy aged 8-months to 8-years-old were enrolled from September (2021-2022) at Iraqi private hospital. Gross Motor Function was assessed by a pediatrician and neurologist specialist, 5 mcg/kg/day of G-CSF subcutaneous single injection daily for three consecutive days. Bone marrow harvested from posterior iliac crest under light general anesthesia. Bone marrow mononuclear cells (BMMNCs) separation was performed using density gradient centrifugation with Ficoll, the cell viability checked by propidium iodide dye in a TALI machine (Invitrogen) in average 98%. The viable BMMNCs injected intrathecal in L4-L5 over a period of 5-10 min. RESULTS: Results: Males accounted for 57.14% (20/35) while female 42.86% (15/35), and main neurological symptoms included spastic disorder spastic disorder (quadriplegia 24 (68.6), tetraplegia 2 (5.7), diplegia 5 (14.28), hemiplegia4 (11.42)). Gross Motor Function Classification System and Gross Motor Function Measure-66 (GMFM-66) showed II 10 (28.58), III 11(31.42) and IV 14 (40). On mean follow-up of 3 months post-stem cell transplant improvement was observed in 80% cases. The improvement showed in gross motor function (6/8) p=0.01, and speech (2/4) p=0.04, neck holding (5/5) p=0.0003, sitting balance (4/4) p=0.04, postural tone (5/5) p=0.0003, as well as significant reduction in seizure frequency (2/3) p=0.04 and improvement in cognition (6/7) p=0.01 were observed. CONCLUSION: Conclusion: Stem cell therapy for cerebral palsy shows a significant positive effect on the gross motor function, without long adverse effects.

Notch1 ligand signaling pathway activated in cervical cancer: poor prognosis with high-level JAG1/Notch1.

OBJECTIVE: Notch signalings are regulated multiple cellular processes during cancer progression. We aimed to investigate the significance and prognostic value of expression of Notch1 and JAG1 in cervical cancer to determine whether they could serve as prognostic predictors. METHODS/MATERIALS: The expression of Notch1/JAGD1 was investigated by real-time PCR, western blot assay and its association with overall survival of patients was analyzed by statistical analysis. RESULTS: Notch1 and JAGD1 expression level were significantly elevated in cervical cancer in comparison to normal specimens and other types of Notch receptors and ligands. It is also proved that Notch1 and JAGD1 expression were to be associated with cervical cancer invasion, lymph node metastasis, and FIGO system. In addition, survival analysis proved that elevated Notch1 and JAGD1 expression were associated with poor overall survival of patients (P = 0.01, P = 0.02 log-rank test), respectively. CONCLUSIONS: The present data proved the over-expression of Notch1/JAGD1 and its association with tumor progression in human cervical cancer, which might be a potential valuable biomarker for cervical cancer and further studies need.

Fibronectin promotes migration and invasion of ovarian cancer cells through up-regulation of FAK-PI3K/Akt pathway.

Ovarian cancer is the leading cause of death from gynecological malignancy, and the fourth most common cause of cancer death among American women. This study investigates the mechanism of fibronectin (FN) in stimulating ovarian cancer cell migration and invasion through up-regulation of focal adhesion kinase (FAK) pathway. Human ovarian cancer cells (OVCAR-3, A2780/CP70) were cultured and treated with fibronectin (10 µg/mL). Trans-well plates were used to conduct the migration assay, real-time RT-PCR for FAK mRNA expression, and FAK siRNA for blocking FAK expression. Western blots were used for P-FAK, P-PI3K, and P-Akt analysis. Fibronectin-treated OVCAR-3, A2780/CP70 cells have increased ability to migrate and invade. It significantly promoted this behavior through the phosphorylation of FAK. The cell displayed significantly increased signaling regulation of the FAK pathway (p-PI3K/P-Akt). Furthermore, siRNA FAK-treated cells had reduced the levels of p-PI3K/P-Akt after induced by fibronectin. Our results indicate that FAK inhibition can suppress ovarian cancer cells migration and invasion through inhibiting downstream signaling (PI3K/AKT), which might be a therapeutic target or biomarker for ovarian cancer.

Expression of IL-32 modulates NF-κB and p38 MAP kinase pathways in human esophageal cancer.

BACKGROUND: Esophageal cancer is the seventh leading cause of cancer death in males in USA, and there is a strong link has been demonstrated between inflammation and esophageal cancer, interleukin (IL)-32 is a recently described pro-inflammatory cytokine characterized by the induction of nuclear factor NF-κB activation, the p38MAPK also plays an important role in key cellular processes related to inflammation and cancer. We investigated whether the IL-32 expression may be involved in esophageal carcinogenesis through modulates the activity of NF-κB and p-p38 MAPK. METHOD: Malignant esophageal tissue and blood samples were obtained from 65 operated untreated patients, normal samples was obtained from 35 patients operated for other reasons as control. IL-32 expression visualized by immunohistochemistry, Real time RT-PCR for IL-32 mRNA expression, NF-κB phosphorylation and phosphorylated p38mapk were analyzed by immunoblotting, ELISA for further detection IL-32 and cytokines (TNF-α, IL-1ß, IL-6 and IL-8) concentration in the patient's sera. RESULTS: IL-32 expression was increased in immunohistochemical staining for malignant esophageal tissue and it's correlated with the relative expression level of IL-32 mRNA P=0.007, the P-NF-κB level elevated in tumor tissue compared with control and no difference in the total NF-κB level P=0.003 while the IL-32 up-regulated the P-pNF-κB in the esophageal tumor P=0.005. There is increase in p-p38MAPK activation underlying IL-32 expression in tumor P=0.004, but no change in total p38 MAPK in malignant esophagus. The plasma level of IL-32 expression was increased in malignant esophageal patients P=0.01, with increased in the levels of the cytokines TNF-α, IL-6, and IL-1ßP<0.05. CONCLUSIONS: Understanding the pathway of IL-32 expression to stimulate the secretion cytokines via the activation of NF-κB and up-regulation of p-p38MAPK may or may not prove to be a therapeutic target, or a biomarker, and future studies will finally answer this hypothesis generated.

Autologous hematopoietic bone marrow and concentrated growth factor transplantation combined with core decompression in patients with avascular necrosis of the femoral head.

The study aimed to assess the effectiveness of autologous hematopoietic bone marrow and concentrated growth factor (CGF) transplantation and core decompression in patients with avascular necrosis of the femoral head (ANFH). We performed a single-center prospective study on 31 patients with non-traumatic early-stage (stage I to III) ANFH based on the 1994 classification of the Association Research Circulation Osseous (ARCO). The patients were subjected to bone marrow aspiration from the posterior iliac crest, separation, and concentration of growth factors from the bone marrow aspirate, core decompression of the femoral head, and injection of hematopoietic bone marrow and CGFs into the necrotic lesion. Patients were evaluated using the visual analogue scale, the WOMAC questionnaire, and X-ray and MRI examinations of the hip joints before, at 2, 4, and 6 months after the intervention. Patients had a mean age of 33 years (range 20-44 years), 19 (61%) of them being male and 12 (39%) females. The presentation of the disease was bilateral in 21 patients and unilateral in 10 patients. The main cause of ANFH was steroid treatment. The mean VAS and WOMAC scores were 48.37 (SD: 14.67) out of 100, and the mean VAS pain score was 50.83 out of 100 (SD: 20.46), respectively, before transplant. This value significantly improved to 22.31 (SD 12.12) of 100, and the mean VAS pain score was 21.31 of 100 (SD: 20.46) (P=0.04). MRI showed a significant improvement (P=0.012). Our results suggest that autologous hematopoietic bone marrow and CGFs transplantation with core decompression have a beneficial effect in early-stage ANFH.

Vitamin E and telmisartan attenuates doxorubicin induced cardiac injury in rat through down regulation of inflammatory response.

BACKGROUND: The importance of doxorubicin (Dox), as a potent antitumor antibiotic, is limited by the development of life-threatening cardiomyopathy. It has been shown that free radicals are involved in acute doxorubicin-induced toxicity. The aim of this study was to determine the protective effect of vitamin E and telmisartan in acute doxorubicin induced cardiotoxicity. METHODS: Thirty two male Sprague - Dawley rats were involved in this study and were randomly separated into 4 groups, eight rats in each group, one group received normal saline I.P as control and second group received doxorubicin 20 mg/kg I.P, the other two groups also received doxorubicin 20 mg/kg I.P as single dose after seven cumulative doses (for seven days) of vitamin E (100 mg/kg) and telmisartan (1 mg/kg) respectively. Immunofluorescent staining for monocytes infiltration and analyses of plasma by (ELISAs) for MCP-1and troponin I. Western immunoblotting assay for ICAM-1, while left ventricular function was analyzed by microcatheter, also estimated the level of oxidative stress parameters (MDA and Catalase) and cardiac enzymes activities (CK-MB and LDH) before starting drugs treatment and after treatment period by 48 hours. RESULTS: The immunofluorescent staining showed that administration of vitamin E and telmisartan are attenuated of mononuclear cell infiltration; (p < 0.05 vs. Dox group), also reduced the level of chemokines MCP-1 and ICAM-1 expression compared with Dox group only, and there is marked reduction of myocardial troponin-I levels with improved LV function in vitamin E and telmisartan treated group. Doxorubicin treatment increased MDA, LDH, CK-MB levels significantly (P < 0.01), and were counteracted by administration of vitamin E and telmisartan, but did not significantly affect serum catalase activity. CONCLUSIONS: Antioxidant effect (vitamin E and telmisartan) have been shown to decrease doxorubicininduced cardiotoxicity.

Novel Toll-like receptor-4 deficiency attenuates trastuzumab (Herceptin) induced cardiac injury in mice.

BACKGROUND: Cardiac inflammation and generation of oxidative stress are known to contribute to trastuzumab (herceptin) induced cardiac toxicity. Toll-like receptors (TLRs) are a part of the innate immune system and are involved in cardiac stress reactions. Since TLR4 might play a relevant role in cardiac inflammatory signaling, we investigated whether or not TLR4 is involved in trastuzumab induced cardiotoxicity. METHODS: Seven days after a single injection of herceptin (2 mg/kg; i.p.), left ventricular pressure volume loops were measured in HeN compotent (TLR4+/+) and HeJ mutant (TLR4-/-) treated with trastuzumab and control mice. Immunofluorescent staining for monocyte infiltration and analyses of plasma by (ELISAs) for different chemokines including: MCP-1and tumor necrosis factor-α (TNF-α), Western immunoblotting assay for ICAM-1, and used troponin I for cardiac injury marker. RESULTS: Trastuzumab injection resulted in an impairment of left ventricular function in TLR-4 competent (HeN), in contrast TLR4-/- trastuzumab mice showed improved left ventricular function EF%, CO; p < 0.05, attenuation of mononuclear cell infiltration in TLR4 -/-; p < 0.05 vs.TLR-4 competent (HeN), reduced level of cytokines TNF-α, MCP-1 and ICAM-1 expression in TLR4-/-, marked reduction of myocardial troponin-I levels in TLR4-deficient mice. Data are presented as means ± SE; n = 8 in each group p < 0.05 vs.TLR-4 competent (HeN). CONCLUSIONS: Treatment with trastuzumab induces an inflammatory response that contributes to myocardial tissue TLR4 mediates chemokine expression (TNF-α, MCP-1and ICAM-1), so in experimental animals TLR4 deficiency improves left ventricular function and attenuates pathophysiological key mechanisms in trastuzumab induced cardiomyopathy.

Indocyanine Green-001 (ICG-001) Attenuates Wnt/ß-catenin-induces Myocardial Injury Following Sepsis.

OBJECTIVE: To investigate the mechanistic pathway of both indocyanine green (ICG)-001 in attenuated endotoxemia-induced cardiac depression through downregulation cardiac Wnt/ ß-catenin cell signaling. MATERIALS AND METHODS: Adult (4-6 months) male Albino-Webster mice, their weights ranged from 25 to 30 g, were pretreated with ICG-001 i.p., following cecal ligation and puncture (CLP). Left ventricle (LV) function was assessed using a microcatheter system. Monocyte chemoattractant protein-1 (MCP-1) and cytokines mediators in plasma and myocardium were analyzed by enzyme-linked immunosorbent assay. Further, the cardiac Wnt protein measured by quantitative real-time polymerase chain reaction while ß-catenin analysis through Western blotting procedure. The pathological changes and cells injury in the myocardium were examined using hematoxylin and eosin staining. RESULTS: CLP mice displayed worse LV function. The exaggerated cardiac depression in CLP mice was associated with higher levels of MCP-1 and cytokines in plasma and myocardium together with greater cardiac levels of cardiac troponin-I and Wnt/ß-catenin. Neutralization of sepsis by either ICG-001resulted in improved LV function and reductions in inflammatory mediators. CONCLUSION: Taken together, these data showed that ICG-001 improved LV function following sepsis through downregulation of Wnt/ß-catenin and serve as a potential mechanistic pathway ICG-001 in therapeutic cardiac endotoxemia in animal model.

Herbal extract targets in Leishmania tropica.

The present study aims to investigate the effect of some herbal extract such as phenolic compounds on the viability of Leishmania tropica promastigotes in vitro. Four tested chemical agents (caffeic acid (CA), ferulic acid (FA), syringic acid (SA) and 4-hydroxybenzoic acid (4-HBA)) were used in this study. The viability of Leishmania tropica promastigotes was investigated under five different concentrations (10, 15, 20, 25 and 30 mg/ml) of each agent after (72 h). CA was the most active agent on the promastigotes viability after 72 h exposure to 30 mg/ml concentration so that the parasiticidal effect reach (53 × 10(4)) promastigote/ml. FA is the second agent in parasiticidal effect that parasiticidal effect reach to (50 × 10(4) promastigote/ml) at a concentration (30 mg/ml), 4-HBA is the third agent in parasiticidal effect that reach to (48 × 10(4) promastigote/ml) at a concentration (30 mg/ml), SA is the weakest agent in parasiticidal activity that reach to (44 × 10(4) promastigote/ml) at a concentration (30 mg/ml). It can be concluded that (CA, FA, SA and 4-HBA) possess acidal effect on the Leishmania tropica promastigotes in vitro.

Role of NF-κß and oxidative pathways in atherosclerosis: cross-talk between dyslipidemia and candesartan.

PURPOSE: The objective of this study is to assess the effect of the candesartan on the progression of atherosclerosis through the downregulation of NF-κß and interference with oxidative pathway. METHODS: Twenty-four rabbits were assigned to three groups: control group fed normal diet; induced untreated group fed 1% cholesterol diet; and treated candesartan group also fed 1% cholesterol diet. Plasma lipid profiles were measured, and ELISA for plasma cytokines and chemokine was performed. Analyses of NF-κß and VCAM-1 were performed using Western blotting with RT-PCR for NF-κB activity at mRNA. Doppler ultrasound was used to evaluate aortic intima-media thickness, and atheroma was detected by H&E staining. Immunofluorescent staining was performed to confirm accumulation of monocytes and PMNs. RESULTS: Candesartan markedly reduced the levels of the plasma lipid profile including total cholesterol [TC], triglycerides [TG], and LDL-C, while significantly elevating levels in the plasma HDL-C, in addition to reducing cytokine (TNF-α, IL-6, IL-1ß) and chemokine levels (MCP-1). Also, it decreased the aortic malondialdehyde (MDA) concentration and elevated the aortic glutathione (GSH) level compared with untreated animals (P < 0.05). The triplex Doppler ultrasound study confirmed that the candesartan attenuated intima-media thickness at 6 months of study. All candesartan-treated rabbits showed significantly attenuated atherosclerosis lesions with reduced accumulation of monocytes and had significantly reduced VCAM-1 expression and NF-κß activity. CONCLUSION: Candesartan retards the progression of atherosclerosis via interference with NF-κß and oxidative pathways.