Adipose-derived stem cells (ASCs) are believed to have potential use for treating many illnesses. Most cells, including ASCs, are generally cultured in medium containing fetal bovine serum (FBS). However, FBS, which could induce an immune response or infection, is not recommended for clinical applications. In the present study, we evaluated the morphology, proliferation rate, and characterization of rabbit ASCs grown in medium containing autologous serum (AS) and compared these cells to ones cultured with FBS. Morphological changes were monitored by microscopy and flow cytometry. Proliferation rates were assessed with cell counting and ASC phenotypes were characterized by flow cytometry using representative surface markers (CD44 and CD45). Expression of epidermal growth factor, brain-derived neurotrophic factor, and vascular endothelial growth factor was measured by reverse transcription-polymerase chain reaction. Results of our study showed that ASCs had a greater expansion rate in AS without developing morphological heterogeneity than cells grown in FBS. AS-cultured ASCs expressed representative growth factors, CD44 but not CD45, similar to cells cultured in FBS. Expression levels of some growth factors were different between AS and FBS. In conclusion, our findings indicated that AS could potentially be used as a culture medium supplement for the expansion of autologous ASCs.
Brain-Derived Neurotrophic Factor ; Bystander Effect ; Cell Count ; Epidermal Growth Factor ; Flow Cytometry ; Intercellular Signaling Peptides and Proteins ; Microscopy ; Phenotype ; Population Characteristics ; Stem Cells ; Vascular Endothelial Growth Factor A

Five monoclonal antibodies (mAbs) that recognize human glutamate dehydrogenase (GDH) have been selected and designated as monoclonal antibodies hGDH60-6, hGDH60-8, hGDH63-10, hGDH63-11, and hGDH91-14. A total of five mAbs recognizing different epitopes of the enzyme were obtained, two of which inhibited human GDH activity. When total proteins of human homogenate separated by SDS- PAGE, were probed with mAbs, a single reactive protein band of 55 kDa, which co-migrated with purified recombinant human GDH was detected. When the purified GDH was incubated with each of the mAbs, its enzyme activity was inhibited by up to 58%. Epitope mapping analysis identified, two subgroups of mAbs recognizing different peptide fragments. Using the individual anti-GDH antibodies as probes, the cross reactivities of brain GDH obtained from human and other animal brain tissues were investigated. For the human and animal tissues tested, immunoreactive bands on Western blots appeared to have the same molecular mass of 55 kDa when hGHD60-6, hGHD60-8, or hGHD91-14 mAbs were used as probes. However, the anti-human GDH mAbs immunoreactive to bands on Western blots reacted differently on the immunoblots of the other animal brains tested, i.e., the two monoclonal antibodies hGDH63-10 and hGDH63-11 only produced positive results for human. These results suggest that human brain GDH is immunologically distinct from those of other mammalian brains. Thorough characterization of these anti-human GDH mAbs could provide potentially valuable tool as immunodiagnostic reagents for the detection, identification and characterization of the various neurological diseases related to the GDH enzyme.
Animals ; Antibodies, Monoclonal/*immunology/isolation & purification ; Cross Reactions ; Electrophoresis, Polyacrylamide Gel ; Glutamate Dehydrogenase/classification/*immunology/isolation & purification ; Human ; Mice ; Organ Specificity ; Rats

OBJECTIVES: Symptoms of attention-deficit hyperactivity disorder (ADHD) during childhood may persist into adulthood. This study included the development and validation process of the Korean Adult ADHD Rating Scale (K-AARS), which was developed for screening and monitoring treatment of adults with ADHD. METHODS: Preliminary questionnaires of the K-AARS were based on the reviews of previous adult ADHD scales and clinical experiences of the board certified child and adolescent psychiatrists in Korea. For this study, 136 adults (18-50 years old) with inattention, hyperactivity and/or impulsivity symptoms were enrolled as ADHD subjects, and compared with 406 control subjects (18-50 years old) without ADHD symptoms. Construct validity was examined using explorative factor analysis and Cronbach's alpha to obtain internal reliability coefficients. Concurrent validity was evaluated by comparison with the Conners' Adult ADHD Rating Scale (CAARS). RESULTS: An explorative factor analysis showed that the K-AARS had 8 factors (inattention, hyperactivity, impulsivity, antisocial personality disorder/conduct disorder/oppositional defiant disorder, impairment, driving, emotional dysregulation, disorganization). K-AARS was highly reliable in terms of internal consistency (Cronbach's alpha 0.77-0.95) and correlation between factors (0.57-0.86). Concurrent validity with the CAARS and discriminant validity were statistically significant. CONCLUSION: The K-AARS is a valid and reliable measure for assessment of Korean adults with ADHD.
Adolescent ; Adult* ; Antisocial Personality Disorder ; Child ; Factor Analysis, Statistical ; Humans ; Impulsive Behavior ; Korea ; Mass Screening ; Psychiatry ; Weights and Measures

Plasminogen activator inhibitor-1 (PAI-1) is a member of serine protease inhibitor family, which regulates the activity of tissue plasminogen activator (tPA). In CNS, tPA/PAI-1 activity is involved in the regulation of a variety of cellular processes such as neuronal development, synaptic plasticity and cell survival. To gain a more insights into the regulatory mechanism modulating tPA/PAI-1 activity in brain, we investigated the effects of proteasome inhibitors on tPA/PAI-1 expression and activity in rat primary astrocytes, the major cell type expressing both tPA and PAI-1. We found that submicromolar concentration of MG132, a cell permeable peptide-aldehyde inhibitor of ubiquitin proteasome pathway selectively upregulates PAI-1 expression. Upregulation of PAI-1 mRNA as well as increased PAI-1 promoter reporter activity suggested that MG132 transcriptionally increased PAI-1 expression. The induction of PAI-1 downregulated tPA activity in rat primary astrocytes. Another proteasome inhibitor lactacystin similarly increased the expression of PAI-1 in rat primary astrocytes. MG132 activated MAPK pathways as well as PI3K/Akt pathways. Inhibitors of these signaling pathways reduced MG132-mediated upregulation of PAI-1 in varying degrees and most prominent effects were observed with SB203580, a p38 MAPK pathway inhibitor. The regulation of tPA/PAI-1 activity by proteasome inhibitor in rat primary astrocytes may underlie the observed CNS effects of MG132 such as neuroprotection.
Animals ; Astrocytes* ; Brain ; Cell Survival ; Humans ; Neurons ; p38 Mitogen-Activated Protein Kinases ; Plasminogen Activator Inhibitor 1 ; Plasminogen Activators* ; Plasminogen* ; Plastics ; Proteasome Endopeptidase Complex ; Proteasome Inhibitors ; Rats* ; RNA, Messenger ; Serine Proteases ; Tissue Plasminogen Activator ; Ubiquitin ; Up-Regulation*