Intrathecal Transplantation of Autologous Adherent Bone Marrow Cells Induces Functional Neurological Recovery in a Canine Model of Spinal Cord Injury.

Spinal cord injury (SCI) results in demyelination of surviving axons, loss of oligodendrocytes, and impairment of motor and sensory functions. We have developed a clinical strategy of cell therapy for SCI through the use of autologous bone marrow cells for transplantation to augment remyelination and enhance neurological repair. In a preclinical large mammalian model of SCI, experimental dogs were subjected to a clipping contusion of the spinal cord. Two weeks after the injury, GFP-labeled autologous minimally manipulated adherent bone marrow cells (ABMCs) were transplanted intrathecally to investigate the safety and efficacy of autologous ABMC therapy. The effects of ABMC transplantation in dogs with SCI were determined using functional neurological scoring, and the integration of ABMCs into the injured cords was determined using histopathological and immunohistochemical investigations and electron microscopic analyses of sections from control and transplanted spinal cords. Our data demonstrate the presence of GFP-labeled cells in the injured spinal cord for up to 16 weeks after transplantation in the subacute SCI stage. GFP-labeled cells homed to the site of injury and were detected around white matter tracts and surviving axons. ABMC therapy in the canine SCI model enhanced remyelination and augmented neural regeneration, resulting in improved neurological functions. Therefore, autologous ABMC therapy appears to be a safe and promising therapy for spinal cord injuries.

Enhancement of Neural Stem Cells after Induction of Depression in Male Albino Rats (A histological & Immunohistochemical Study).

BACKGROUND AND OBJECTIVES: Depression is one of the most prevalent psychiatric disorders. Endogenous neural stem cells (NSCs) could replace damaged Hippocampal neurons in depression. This work was planned to evaluate Rhodiola rosea (Rr) extract possible role in stimulation of NSCs proliferation and in depression improvement. METHODS AND RESULTS: Thirty adult male albino rats were divided into three groups; control, untreated depressed model and Rr model. After depression induction by chronic mild stress, rats received Rr extract 1.5 g/kg/day for three weeks. The sucrose preference test (SP) was done before, after depression induction and 3 weeks after supplementation of Rr. The brain was removed and processed for H&E and immunohistochemical staining for caspase 3, glial fibrillary acid protein (GFAP) and proliferating cell nuclear antigen (PCNA). Rr group revealed improved sucrose preference, increased undamaged neurons and decreased dark neurons. Moreover, Caspase 3 +ve cells were not detected, GFAP +ve cells increased and PCNA +ve cells were detected only in Rr group. CONCLUSIONS: This work points to the role of Rr in depression improvement and in stimulation of NSCs proliferation.

Myelo-enhancement by astragalus membranaceus in male albino rats with chemotherapy myelo-suppression. Histological and immunohistochemical study.

BACKGROUND AND OBJECTIVES: Myelosuppression is the most common toxicity encountered in the oncology clinic today. This study was planned to investigate the possible protective and therapeutic role of the traditional Chinese Medicinal Herb; Astragalus Membranaceus (AM), on chemotherapy-induced myelosuppression. METHODS AND RESULTS: This study was carried out on thirty six adult male albino rats. They were divided into: Group I Control Group (n=6) received a vehicle of phosphate buffered saline (PBS) solution. Group II (n=12) were injected I.P. with cyclophosphamide (CY) for 3 days (gIIa n =6) and continued for one more week to receive AM orally (gIIb n=6). Group III (n=6) received CY I.P. together with AM orally for 3 days. Group IV (n=12) received AM orally for one week (gIVa n=6) and continued for extra three days receiving CY I.P. with AM orally (gIVb n=6). Blood samples were analysed for Total Leucocytic Count and Lymphocytic Count. Counting of CD34 +ve cells in bone marrow was performed by flowcytometry. Bone marrow sections were subjected to H&E stain as well as immunohistochemical staining for anti- CD20 antibody. The mean area % of cellular bone marrow regions occupied by developing haemopoietic cells, mean area of fat cells and mean number of CD20 immunopositive B lymphocytes in the bone marrow were measured by histomorphometric studies and statistically compared. AM proved to have a myelo-protective and myelo-therapeutic capacity, evidenced at both laboratory and morphological levels. CONCLUSIONS: The greatest myelo-potentiating effect of AM was achieved when supplied before and together with CY therapy.

The effect of in vivo mobilization of bone marrow stem cells on the pancreas of diabetic albino rats (a histological & immunohistochemical study).

BACKGROUND AND OBJECTIVES: The rapidly increasing number of diabetic patients across the world drew the attention to develop more effective therapeutic approaches. Recent investigations on newly differentiated insulin producing cells (IPCs) revealed that they could be derived from embryonic, adult mesenchymal and hematopoietic stem cells. This work was planned to evaluate the role of StemEnhance (Aphanizomenon flos-aquae [AFA] plant extract) in mobilizing naturally occurring bone marrow stem cells as well as in improving streptozotocin-induced diabetic rats. METHODS AND RESULTS: Twenty adult male albino rats were divided into four groups namely the control, the diabetic, the positive control-StemEnhance and the diabetic-StemEnhance groups. After diabetes induction by streptozotocin (STZ), rats received StemEnhance for four weeks. The mean number of blood CD34 immunopositive cells was measured by flowcytometry and random blood sugar was measured weekly. The pancreas was removed from the sacrificed rats and processed for staining with H&E and immunohistochemical staining for CD34+ve and insulin +ve cells. CD34+ve cells increased in the blood after introduction of StemEnhance. CD34+ve cells were observed in the pancreas and the insulin producing cells in the islets of Langerhans were increased from the second to the fourth week of treatment. Blood glucose level improved but it was still higher than the control level after four weeks of StemEnhance treatment. CONCLUSIONS: This work points to the significant role of StemEnhance in stem cell mobilization and the improvement of diabetes mellitus.

Novel antitumor acetamide, pyrrole, pyrrolopyrimidine, thiocyanate, hydrazone, pyrazole, isothiocyanate and thiophene derivatives containing a biologically active pyrazole moiety.

4-Amino-N-(1-phenyl-1H-pyrazol-5-yl)-benzenesulfonamide (sulfaphenazole) 1 was selected as strategic starting material for the synthesis of some novel acetamide 2, pyrrole 4, pyrrolo[2,3-d]pyrimidine 5, thiocyanate 6, hydrazone 7a,b pyrazole 8, isothiocyanate 9 and thiophene 12 derivatives to evaluate theantitumor activity. Compound 4 was more effective than the reference drug, doxorubicin (CAS 23214-92-8) as positive control.

cDNA cloning and spatiotemporal expression during avian embryogenesis of hnRNP A1, a regulatory factor in alternative splicing.

Alternative splicing and selective transport of RNA transcripts from cell nuclei are important regulatory mechanisms of gene expression during embryonic development. Here we report the molecular characterization and developmental expression in several tissue and organ systems of chicken hnRNP A1, a nucleo-cytoplasmic 'shuttle' protein which in mammalian systems has been shown to function in the regulation of RNA alternative splicing by antagonizing constitutive splicing factors such as SF2/ASF. We show that hnRNP A1 is represented in the chicken by a single gene which is widely expressed at early embryonic stages, with particularly high levels of expression in the brain, skin, developing gut, and other ectodermal and endodermal derivatives. At later stages, expression of its mRNA and protein product become progressively confined to specific organ primordia and cell types, where both transient and persistent expression patterns are observed. HnRNP A1 protein is expressed at sites of active neurogenesis in the developing central and peripheral nervous systems, regions of known extensive alternative splicing.