Association between delayed recall and T2* relaxation time of the subiculum in adolescents: Implications for ultra-high-field magnetic resonance imaging.

AIM: The aim of this study was to assess neuropsychological correlations with the T2* relaxation time (T2*-RT) of hippocampal subregions in adolescents using ultra-high-field (UHF) 7.0-T magnetic resonance imaging (MRI). METHODS: We assessed the T2*-RT of hippocampal subregions in 31 healthy 11th- or 12th-grade high school students using an UHF 7.0-T MRI system. T2*-RT of the cornu ammonis (CA) 1, CA2, CA3, and CA4 subregions and the subiculum were calculated for both the left and right hippocampus. Seven subtests of the Cambridge Neuropsychological Test Automated Battery were administered to the subjects to assess visuospatial memory. RESULTS: Poor performances in delayed recall in the pattern-recognition test were significantly correlated with longer T2*-RT in the bilateral subiculum (right, r = -0.480, P = 0.006; left, r = -0.648, P < 0.001) and the left CA2 (r = -0.480, P = 0.006). CONCLUSION: This study showed that longer T2*-RT in the subiculum were associated with poorer performances in delayed recall in the visual memory tasks. This finding suggests that the subiculum might play a predominant role in delayed recall in adolescents.

Quantitative analysis of the hippocampus using images obtained from 7.0 T MRI.

In-vivo volumetric measurements of hippocampus have proven to be highly informative for studying various neurological diseases such as Alzheimer's disease. The usefulness of volumetric imaging, however, has been limited due to the poor image resolutions obtained by currently available MRI images. In this study, a new result of volumetric image measurement of the hippocampus using 7.0 T MRI images of high contrast and resolution is described. To verify the usefulness of the proposed method, its reliability and sensitivity were examined and compared with existing imaging techniques such as 1.5 T or 3.0 T MRI imaging. The results of our study with 7.0 T MRI clearly demonstrated superior boundary detection for the hippocampal head, body, and tail compared with low field MRIs. In conclusion, robust and reproducible volumetric measurements as well as 3D images of clear contrast obtained with 7.0 T suggest the usefulness of high field MR imaging and its eventual use for the accurate diagnosis of hippocampal diseases and related research.

Tat-mediated protein transduction of human brain pyridoxine-5-P oxidase into PC12 cells.

Pyridoxine-5-P oxidase catalyses the terminal step in the biosynthesis of pyridoxal-5-P, the biologically active form of vitamin B6 which acts as an essential cofactor. Here, a human brain pyridoxine-5-P oxidase gene was fused with a gene fragment encoding the HIV-1 Tat protein transduction domain (RKKRRQRRR) in a bacterial expression vector to produce a genetic in-frame Tat-pyridoxine-5-P oxidase fusion protein. Expressed and purified Tat-pyridoxine-5-P oxidase fusion protein transduced efficiently into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced Tat-pyridoxine-5-P oxidase protein showed catalytic activity and was stable for 48 h. Moreover, the formation of pyridoxal-5-P was increased by adding exogenous Tat-pyridoxine-5-P oxidase to media pre-treated with the vitamin B6 precursor pyridoxine. In addition, the intracellular concentration of pyridoxal-5-P was markedly increased when Tat-pyridoxal kinase was transduced together with Tat-pyridoxine-5-P oxidase into cells.These results suggest that the transduction of Tat-pyridoxine-5-P oxidase fusion protein presents a means of regulating the level of pyridoxal-5-P and of replenishing this enzyme in various neurological disorders related to vitamin B6.

Transduced Tat-alpha-synuclein protects against oxidative stress in vitro and in vivo.

Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of alpha-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant alpha-synuclein genes were fused with a gene fragment encoding the nine amino acid transactivator of transcription (Tat) protein transduction domain of HIV-1 in a bacterial expression vector to produce a genetic in-frame WT Tat-alpha-synuclein (wild type) and mutant Tat-alpha-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-alpha-synucleins in vitro and in vivo. WT Tat-alpha-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-alpha-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-alpha-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-alpha-synuclein. These results suggest that transduced Tat-alpha-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.

Transduced human copper chaperone for Cu,Zn-SOD (PEP-1-CCS) protects against neuronal cell death.

Reactive oxygen species (ROS) contribute to the development of various human diseases. Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS. SOD activity is dependent upon bound copper ions supplied by its partner metallochaperone protein, copper chaperone for SOD (CCS). In the present study, we investigated the protective effects of PEP-1-CCS against neuronal cell death and ischemic insults. When PEP-1-CCS was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Moreover, transduced PEP-1-CCS markedly increased endogenous SOD activity in the cells. Immunohistochemical analysis revealed that it prevented neuronal cell death in the hippocampus in response to transient forebrain ischemia. These results suggest that CCS is essential to activate SOD, and that transduction of PEP-1-CCS provides a potential strategy for therapeutic delivery in various human diseases including stroke related to SOD or ROS.

Human brain pyridoxal-5'-phosphate phosphatase: production and characterization of monoclonal antibodies.

We cloned and expressed human pyridoxal-5\'-phosphate (PLP) phosphatase, the coenzymatically active form of vitamin B6, in Escherichia coli using pET15b vector. Monoclonal antibodies (mAb) were generated against purified human brain PLP phosphatase in mice, and four antibodies recognizing different epitopes were obtained, one of which inhibited PLP phosphatase. The binding affinities of these four mAbs to PLP phosphatase, as determined using biosensor technology, showed that they had similar binding affinities. Using the anti-PLP phosphatase antibodies as probes, we investigated their cross-reactivities in various mammalian and human tissues and cell lines. The immunoreactive bands obtained on Western blots had molecular masses of ca. 33 kDa. Similarly fractionated extracts of several mammalian cell lines all produced a single band of molecular mass 33 kDa. We believe that these PLP phosphatase mAbs could be used as valuable immunodiagnostic reagents for the detection, identification, and characterization of various neurological diseases related to vitamin B6 abnormalities.

Altered T2* relaxation time of the hippocampus in major depressive disorder: implications of ultra-high field magnetic resonance imaging.

Previous studies with 1.5 T or 3.0 T magnetic resonance imaging (MRI) have produced mixed results regarding the structural changes of the hippocampus in major depressive disorder (MDD). Subtle region-specific hippocampal tissue changes might be more sensitively detected by measuring the T2* relaxation time (T2*-RT) by ultra-high-field (UHF) MRI, as it provides much higher contrast and sensitivity and consequently greater resolution. We assessed the T2*-RTs of hippocampal sub-regions in 16 MDD patients (9 with recurrent MDD) and 16 control subjects using an UHF 7.0 T MRI system. T2*-RTs of CA1, CA2, CA3, CA4, and subiculum were calculated for both left and right hippocampus. MDD patients had significantly longer T2*-RTs in the right CA1 and subiculum than control subjects. Patients with recurrent MDD had significantly longer T2*-RTs in the right subiculum than those experiencing a first depressive episode, and longer T2*-RTs in the right CA1, CA3, and subiculum than control subjects. Values for T2*-RTs of the right CA3 were significantly correlated with illness duration. In conclusion, we report that T2*-RTs in the right subiculum and CA1 were increased in patients with MDD, especially in cases of recurrent MDD. These findings suggest that region-specific hippocampal damage may be occurring in recurrent depression.