Cytoskeleton disruption causes apoptotic degeneration of dentate granule cells in hippocampal slice cultures

Colchicine, a potent microtubule-depolymerizing agent, is well known to selectively kill dentate granule cells in the hippocampal formation in vivo. Using organotypic cultures of rat entorhino-hippocampal slices, we confirmed that in vitro exposure to 1 μM and 10 μM of colchicine reproduced a specific degeneration of the granule cells after 24 h. Similar results were obtained with other types of microtubule-disrupting agents, i.e., nocodazole, vinblastine, and Taxol. Interestingly, the actin-depolymerizing agents cytochalasin D and latrunculin A also elicited selective neurotoxicity in the dentate gyrus without affecting survival of hippocampal pyramidal cells. The selective pattern of degeneration was observable 24 h after a brief treatment with the toxins as short as 5 min, but this delayed neuronal death was unlikely to be a result of excitotoxicity because it was virtually unaffected by glutamate receptor antagonists, tetrodotoxin, or extracellular Ca²⁺-free conditions. The damaged tissues contained a large number of TUNEL-positive neurons and exhibited an increased level in caspase-3-like activity, suggesting that cytoskeleton disruption triggers an apoptosis-like process in dentate granule cells. Thus, this study may provide a basis for understanding the distinctive mechanism that supports granule cell survival.     

Skin problems among fiber-glass reinforced plastics factory workers in Japan

Two surveys, one in winter the other in summer time, examined the skin problems of the entire manual workers (N=148) from 11 small-to-medium sized fiber-glass reinforced plastics (FRP) factories located in Kyushu, Japan. The workers were exposed to unsaturated polyester resin, including styrene and auxiliary agents such as cobalt naphthenate, hardeners such as methyl ethyl ketone peroxides, glass fiber and dust including shortened glass fiber and plastic particles. Eighty-seven workers (58.8%) reported having skin problems (mainly itching or dermatitis) since they started to work in FRP manufacturing and 25 workers had consulted a physician because of their skin problems; one worker was forced to take sick leave because of his severe dermatitis. History of allergic diseases and shorter occupational period (duration of employment) in a FRP factory were associated with greater probability of having a history of work-related skin symptoms. Workers in factories where dust-generating and lamination sites were located in different buildings were significantly less likely to have a history of skin problems than those in factories where the two sites were located in the same building. Of the 67 workers examined in both seasons closed to double the prevalence of dermatitis was found in summer (23.3%) than winter (13.4%).     

Evaluation of an exercise program for obese males

PURPOSE: We developed a one-year exercise program based on evaluation of life style and physical fitness and investigated its efficacy. SUBJECTS AND METHODS: Sixty one Japanese males (30 < or = age < 59, body mass index: BMI > or = 26.4 kg/m2) were enrolled in the stady. The exercise program consisted of four sections: (1) Preparation for preventing injury; (2) Increase of daily activity; (3) Enjoying exercise; (4) Preparation and practice of individual-based on a exercise program. The subjects visited Okayama Southern Institute of Health and were being monitored weekly for one-year. They were instructed to check daily steps every day and increase daily walking to at least 1000 steps more than the daily walk at baseline. Before and after one-year follow up study, we evaluated life style and physical fitness in terms of body composition, aerobic exercise level, muscle strength and flexibility. RESULTS: The continuation rate for the program was 66%. Body weight (pre 81.5 +/- 7.4 kg vs post 78.6 +/- 7.3 kg), body fat percentage (pre 30.3 +/- 4.1% vs post 28.4 +/- 5.0%) and waist hip ratio (pre 0.95 +/- 0.04 vs post 0.92 +/- 0.04) were significantly reduced. Aerobic exercise level and muscle strength were increased. Regarding volition for exercise, execution, continuation and steps per day were also improved. CONCLUSION: Regular exercise at an institution (once a week) and change in daily life style may play critical role in reducing body weight. This new program proved quite useful for Japanese obese males.     

Voxel-based comparison of regional cerebral glucose metabolism between PSP and corticobasal degeneration

OBJECTIVES: Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are neurodegenerative disorders that may be accompanied by dementia and parkinsonism as clinical symptoms. The purpose of this study was to elucidate cerebral metabolic differences of these two diseases with cognitive impairments by [18F] fluorodeoxyglucose (FDG) and positron emission tomography (PET). METHODS: A total of 12 patients with PSP (age: 62.8+/-6.0 years old, m: 7, f: 5, Mini-Mental State Examination (MMSE): 23.4+/-2.6), 12 patients with CBD (age: 64.8+/-6.3 years old, m: 6, f: 6, MMSE: 22.9+/-4.5), and age-matched healthy subjects (normal control (NC)) (age: 63.8+/-7.7 years old, m: 7, f: 5) were subjected to FDG-PET to obtain glucose metabolic images. We compared regional cerebral metabolic images by a voxel-by-voxel analysis with statistical parametric mapping (SPM) among PSP, CBD, and NC subjects, and evaluated differences of hypometabolic regions. RESULTS: The patients with PSP showed reduced cerebral glucose metabolism in the medial and lateral frontal gyri, basal ganglia, and midbrain compared with NC, whereas the patients with CBD showed significant reduction in the parietal lobes (p<0.001). SPM also revealed parietal hypometabolism in CBD patients compared with PSP patients (p<0.001). CONCLUSIONS: The predominant parietal glucose metabolic reduction in CBD patients was different from previously reported findings. This finding would be the characteristic substance of patients with CBD accompanying cognitive impairments. Our findings suggest that measurement of glucose metabolism by PET and a voxel-based analysis is useful to understand the pathophysiology of these two diseases with cognitive impairments.     

Variation in FDG uptakes in different regions in normal human brain as a function of the time (30 and 60 minutes) after injection of FDG

OBJECTIVE: The authors' goal was to determine whether FDG uptakes in various regions of the brain are different for early and late scanning time in positron emission tomography (PET). METHOD: F-18 fluorodeoxyglucose (FDG) PET was performed on 15 healthy normal subjects to obtain early and late acquisition glucose metabolic images (30 and 60 min after FDG injection), respectively. The two sets of images were compared in a voxel-by-voxel analysis. RESULTS: In the bilateral posterior cingulate gyrus, parietal and frontal association cortices, and subcallosal cortices, the FDG uptakes were larger on the late scan image than on the early scan image, and the FDG uptakes were larger in the cerebellar hemisphere, vermis and frontal basis on the early scan image than on the late scan image. CONCLUSIONS: These results suggest that there are different regional FDG uptakes depending on the scanning time after FDG injection and we must be careful in replacing conventional FDG PET scanning with early scanning in FDG PET study.     

Antitumor effects on mouse melanoma elicited by local secretion of interleukin-12 and their enhancement by treatment with interleukin-18

To investigate the mechanism of the antitumor effect of locally secreted interleukin-12 (IL-12), we introduced the IL-12 p35 and p40 cDNAs into mouse B16 melanoma cells. IL-12 gene-transfected B16 melanoma (B16/IL12) showed marked retardation of tumor growth when implanted subcutaneously into syngeneic mice. In these mice, depletion of not only Natural Killer (NK) cells but also CD8+ T cells diminished the antitumor effect of locally secreted IL-12. Immunohistochemical analysis showed that NK cells and macrophages accumulated more densely at the center and periphery of B16/IL12 tumors than that of parental B16 tumors, whereas CD4+ T cells and CD8+ T cells accumulated sparsely only at the periphery of both transfected and untransfected tumors. Systemic treatment with interleukin-18 (IL-18) markedly inhibited the growth of B16/IL12 but did not influence the tumor growth of parental B16 cells in vivo. These results suggest that local IL-12 secretion can retard the growth of B16 melanoma mediated primarily by NK cells and indirectly by CD8+ T cells and that its antitumor effect is augmented by systemic treatment with the novel cytokine IL-18.     

Roles for the protein tyrosine phosphatase SHP-2 in cytoskeletal organization, cell adhesion and cell migration revealed by overexpression of a dominant negative mutant

SHP-2, a SRC homology 2 domain-containing protein tyrosine phosphatase, mediates activation of Ras and mitogen-activated protein kinase by various mitogens and cell adhesion. Inhibition of endogenous SHP-2 by overexpression of a catalytically inactive (dominant negative) mutant in Chinese hamster ovary cells or Rat-1 fibroblasts has now been shown to induce a marked change in cell morphology (from elongated to less polarized) that is accompanied by substantial increases in the numbers of actin stress fibers and focal adhesion contacts. Overexpression of the SHP-2 mutant also increased the strength of cell-substratum adhesion and resulted in hyperphosphorylation of SHPS-1, a substrate of SHP-2 that contributes to cell adhesion-induced signaling. Inhibition of SHP-2 also markedly increased the rate of cell attachment to and cell spreading on extracellular matrix proteins such as fibronectin and vitronectin, effects that were accompanied by enhancement of adhesion-induced tyrosine phosphorylation of paxillin and p130Cas. In addition, cell migration mediated by fibronectin or vitronectin, but not that induced by insulin, was impaired by overexpression of the SHP-2 mutant. These results suggest that SHP-2 plays an important role in the control of cell shape by contributing to cytoskeletal organization, and that it is an important regulator of integrin-mediated cell adhesion, spreading, and migration as well as of tyrosine phosphorylation of focal adhesion contact-associated proteins.