Apoptosis induction by wheat-flour sphingoid bases in DLD-1 human colon cancer cells

The apoptotic effects of plant sphingoid bases prepared from wheat-flour cerebroside on human colorectal cancer DLD-1 cells were examined. The viability of DLD-1 cells treated with such plant sphingoid bases was reduced in a dose-dependent manner and was similar to that of cells treated with sphingosine. Morphological changes such as condensed chromatin fragments were found, so those sphingoid bases reduced cell viability through causing apoptosis in these cells.     

Histochemical analysis of macrophage migration inhibitory factor in psoriasis vulgaris

Psoriasis is a persistent cutaneous disease characterized by skin inflammation and infiltration of immunocytes such as lymphocytes and monocytes/macrophages, concomitant with abnormal epidermal hyperproliferation. We previously showed that the serum level of macrophage migration inhibitory factor (MIF) and its production by peripheral blood mononuclear cells of patients with psoriasis were closely correlated with the severity of clinical symptoms; however, the precise role of MIF in psoriatic epidermis remains to be clarified. The current study was carried out to elucidate the possible involvement of MIF in psoriasis, using immunohistochemistry and in situ hybridization. In contrast to elevated serum MIF in psoriasis, MIF-positive staining in the lesional psoriatic epidermis was significantly decreased, as demonstrated by immunohistochemical analysis using an anti-MIF antibody. Consistent with this finding, we found, by in situ hybridization, that MIF mRNA concomitantly decreased in the psoriatic lesions. Although the reason for the different MIF levels in the psoriatic epidermis and in the circulation remains unknown, it is hypothesized that MIF, a potential growth factor, might be decreased in psoriatic lesions to counterregulate the abnormal epidermal proliferation caused by dysregulation of cytokines and growth factors.     

Role of macrophage migration inhibitory factor (MIF) in peripheral nerve regeneration: anti-MIF antibody induces delay of nerve regeneration and the apoptosis of Schwann cells

BACKGROUND: Macrophage migration inhibitory factor (MIF) is a pluripotent cytokine involved in inflammation and immune responses as well as in cell growth. Although we previously demonstrated the presence of MIF in peripheral nerves, and MIF mRNA expression was up-regulated after axotomy, the role of MIF in nerve injury and regeneration has not been evaluated. MATERIALS AND METHODS: To examine the potential role of MIF in nerve regeneration, we locally administered an anti-MIF polyclonal antibody into regenerating rat sciatic nerves using the silicone chamber model. The effect of the anti-MIF antibody on nerve regeneration was evaluated using an axonal reflex test. In addition, we carried out a terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) assay and immunohistochemical analysis of the damaged nerve segments with regard to apoptosis-related proteins such as p53 to evaluate the effects of anti- MIF antibodies on apoptosis during the regeneration process. RESULTS: The regeneration length of the nerve in the anti-MIF antibody-treated group was significantly shorter than that in the non-immune rabbit IgG-treated group at weeks 2, 4 and 6 after surgery. TUNEL assay showed that a large number of apoptotic cells, mostly Schwann cells, were observed in the intratubal and distal nerve segments at weeks 4 and 6 after surgery by the anti-MIF antibody treatment. Consistent with these results, Ki-67-positive cells were significantly decreased by the anti-MIF antibody treatment. Immunohistochemical analyses revealed that p53 and, to a lesser extent, Fas were more up-regulated in the anti-MIF antibody-treated nerves than in the controls. CONCLUSION: Taken together, these results suggest that MIF plays an important role in acceleration of peripheral nerve regeneration and in prevention of Schwann cell apoptosis, mainly through overcoming the apoptotic effect of p53.     

Receptors for recombinant feline interferon-omega in hemocytes of the Japanese pearl oyster Pinctada fucata martensii

In a previous study we determined that administration of recombinant feline interferon-omega (rFeIFN-omega) could protect Japanese pearl oysters Pinctada fucata martensii from akoya-virus infection. Our results suggested that rFeIFN-omega enhanced the potential of agranulocytes to phagocytize necrotic cells and to produce collagen fibers that repair the lesions caused by the akoya-virus. In the present study, using an indirect immunofluorescence technique with an anti-rFeIFN-omega rabbit serum, we examined whether hemocytes bear receptors that bind rFeIFN-omega. rFeIFN-omega receptors were present on agranulocytes and bound rFeIFN-omega, and appeared as green fluorescent spots in the cytoplasm under a fluorescent microscope. Around 56% of agranulocytes in Japanese pearl oysters bore the rFeIFN-omega receptors.     

Changes in center-of-pressure dynamics during upright standing related to decreased balance control in young adults: fractional Brownian motion analysis

We investigated the relationships between the ability to maintain balance in an upright stance and center-of-pressure (COP) dynamic properties in young adults. Included in this study were 10 healthy male subjects in each of two groups with respect to balance ability. Balance ability was evaluated according to the length of time a subject stood on one leg with his eyes closed. The means and ranges of this one-leg balancing time were 17.9 s (3-43 s) and 118.3 s (103-120 s) for the off-balance and balance groups, respectively. The time-varying displacements of the COP under a subject's feet during quiet two-leg (normal) standing were measured by an instrumented force platform. Each subject was tested in both the eyes-open and eyes-closed conditions. The COP trajectories were analyzed as fractional Brownian motions according to the procedure of 'stabilogram-diffusion analysis', proposed by Collins and De Luca (1993). The extracted parameters were the effective diffusion coefficients (D) for the short-term (less than about 1.0 s) and long-term intervals, respectively, as well as the Hurst exponents (H) for the short-term and long-term intervals, and some critical-point coordinates (i.e., critical mean square displacements and critical time intervals). The off-balance group showed significantly higher values for short-term D, short-term H, and critical mean square displacements than the balance group. No significant differences between the groups were found in the long-term D and H or in the critical time intervals. That is, for the off-balance subjects, an increase in the stochastic activity and positively correlated (persistent) behavior of the postural sway during shorter timescales may cause postural instability. These results suggest that the difference in balance ability for young adults is related to the open-loop (i.e., short-term) control mechanisms but not to the corrective feedback (i.e., long-term) mechanisms used to maintain balance in an upright stance.     

Diagnostic evaluation of 2', 5'-oligoadenylate synthetase activities and antibodies against Epstein-Barr virus and Coxiella burnetii in patients with chronic fatigue syndrome in Japan

To investigate the association of viral infections with chronic fatigue syndrome (CFS), we assayed 2', 5'-oligoadenylate synthetase (2-5AS) activities in peripheral blood mononuclear cells from CFS patients in Japan. These patients were diagnosed in two hospitals, H1 and H2, located in different areas of the country. The activities were detected in 19 (86%) and 7 (32%) of each of the 22 patients in H1 and H2, respectively, while they were detected in only four (11%) out of the 38 healthy controls. IFN-alpha was similarly detected in a few CFS patients and healthy controls. We also assayed the antibody titers against Epstein-Barr virus (EBV) and Coxiella burnetii in these patients. The EBV anti-EA-IgG antibodies were detected in two (9%) and seven (32%) of each of the 22 patients in H1 and H2, respectively. Anti-C. burnetii IgG antibodies were detected in six (27%) out of 22 patients in H1 but not in 22 patients in H2, while they were detected in one (11%) of the nine healthy controls. Some CFS patients may be associated with EBV or C. burnetii infection. There were some statistical correlations between the 2-5AS activities and antibody titers of EA-IgG (P < 0.05, Student's t-test) but not to the antibody titers of C. burnetii. The up-regulation of 2-5AS activities suggests immunological dysfunctions with some virus infections in the CFS patients. Our results indicate that 2-5AS activities are useful for a diagnostic marker of CFS and for exploring the complicated pathogenesis of CFS.     

Synthesis of a callosic substance during rhizoid differentiation in Spirogyra

Spirogyra living in running water is anchored to the substratum by rhizoids that form at the ends of the filaments. A new terminal cell differentiates into a rhizoid cell if the filament is injured. The mode of growth changes from diffuse to tip growth when rhizoid differentiation begins. In this study, we found that a callosic substance was synthesized during rhizoid differentiation. Decreasing the cell turgor, lowering extracellular Ca2+ or adding Gd3+, all inhibited the commencement of rhizoid differentiation as well as synthesis of the callose-like substance at the tip of the terminal cell. A callosic substance was also synthesized during formation of the conjugation tube.     

The antioxidant cocktail,effective microorganism X (EM-X), protects retinal neurons in rats against N-methyl-D-aspartate excitotoxicity in vivo

Injection of the glutamate agonist N-methyl-D-aspartate (NMDA) into the vitreous body of rats resulted in severe degeneration of neurons in the retina, with a loss of 81% of ganglion cells and 43% of non-ganglion cells. The cocktail EM-X is a novel antioxidant drink derived from ferment of unpolished rice, papaya and sea-weeds with effective microorganisms (EM-X). In animals treated with an intraperitoneal injection of EM-X, the loss of ganglion cells was reduced to 55% and that of non-ganglion cells to 34% when compared to untreated NMDA-injected retinas. Cell degeneration resulting from NMDA excitotoxicity, is thought to be mediated via oxidative stress mechanisms. The neuroprotective effect of the EM-X in this system is therefore likely to be due, at least in part, to its flavonoids, saponins, vitamin E and ascorbic content.