Transient Brain Ischaemia Provokes Ca2+, PIP2 and Calpain Responses Prior to Delayed Neuronal Death in Monkeys

To clarify the mechanism of postischaemic delayed cornu Ammonis (CA)-1 neuronal death, we studied correlations among calpain activation and its subcellular localization, the immunoreactivity of phosphatidylinositol 4,5-bisphosphate (PIP₂) and Ca²⁺ mobilization in the monkey hippocampus by two independent experimental approaches: in vivo transient brain ischaemia and in vitro hypoxia-hypoglycaemia of hippocampal acute slices. The CA-1 sector undergoing 20 min of ischaemia in vivo showed microscopically a small number of neuronal deaths on day 1 and almost global neuronal loss on day 5 after ischaemia. Immediately after ischaemia, CA-1 neurons ultrastructurally showed vacuolation and/or disruption of the lysosomes. Western blotting using antibodies against inactivated or activated μ-calpain demonstrated μ-calpain activation specifically in the CA-1 sector immediately after ischaemia. This finding was confirmed in the perikarya of CA-1 neurons by immunohistochemistry. CA-1 neurons on day 1 showed sustained activation of μ-calpain, and increased immunostaining for inactivated and activated forms of μ- and m-calpains and for PIP₂. Activated μ-calpain and PIP₂ were found to be localized at the vacuolated lysosomal membrane or endoplasmic reticulum and mitochondrial membrane respectively, by immunoelectron microscopy. Calcium imaging data using hippocampal acute slices showed that hypoxia-hypoglycaemia in vitro provoked intense Ca²⁺ mobilization with increased PIP₂ immunostaining specifically in CA-1 neurons. These data suggest that transient brain ischaemia increases intracellular Ca²⁺ and PIP₂ breakdown, which will activate calpain proteolytic activity. Therefore, we suggest that activated calpain at the lysosomal membrane, with the possible release of biodegrading enzyme, will cause postischaemic CA-1 neuronal death.     

Epstein-Barr Virus-positive Intestinal Diffuse Large B-cell Lymphoma in a Japanese Patient with Celiac Disease: First Reported Case and a Literature Review

A 60-year-old Japanese woman was diagnosed with celiac disease (CeD) and treated with a gluten-free diet. For five years, she had a good clinical course. However, she complained of inappetence and nausea. Colonoscopy revealed ulcerative tumors in the terminal ileum. A histological examination of biopsy specimens from the ulcerative tumor showed diffuse infiltration of large atypical lymphocytes. Immunohistologically, the atypical lymphoid cells were positive for cluster of differentiation (CD)10 and CD20. Many Epstein-Barr virus-encoded small RNA (EBER)-positive atypical lymphocytes were detected by in situ hybridization. This represents the first reported case of Epstein-Barr virus-positive intestinal diffuse large B-cell lymphoma complicated with CeD.     

Hypothermia inhibits translocation of CaM kinase II and PKC-alpha, beta, gamma isoforms and fodrin proteolysis in rat brain synaptosome during ischemia-reperfusion

To clarify the involvement of intracellular signaling pathway and calpain in the brain injury and its protection by mild hypothermia, immunoblotting analyses were performed in the rat brain after global forebrain ischemia and reperfusion. After 30 min of ischemia followed by 60 min of reperfusion, Ca2+/calmodulin-dependent kinase II (CaM kinase II) and protein kinase C (PKC)-alpha, beta, gamma isoforms translocated to the synaptosomal fraction, while mild hypothermia (32 degrees C) inhibited the translocation. The hypothermia also inhibited fodrin proteolysis caused by ischemia-reperfusion, indicating the inhibition of calpain. These effects of hypothermia may explain the mechanism of the protection against brain ischemia-reperfusion injury through modulating synaptosomal function.     

Primate neurons show different vulnerability to transient ischemia and response to cathepsin inhibition

Previously, we reported "calpain-induced leakage of lysosomal enzyme cathepsin" as a mechanism of ischemic neuronal death specific for primates. Cathepsin inhibitors such as CA-074 and E-64c were demonstrated to significantly inhibit hippocampal neuronal death. Pyramidal neurons of the hippocampus, Purkinje cells in the cerebellum, and neurons in the caudate nucleus, outer putamen and cortical III, V layers, are known to be vulnerable to ischemia. However, regional differences of the vulnerability and response to neuroprotectants, have not been studied in detail. Here, the monkey brains undergoing transient ischemia were studied to clarify such regional differences by the microscopic counting of surviving neurons. The dead neurons were characterized by eosinophilic coagulation necrosis without apoptotic bodies. The control postischemic brain without treatment showed surviving neurons in caudate nucleus (55.8%), outer putamen (44.4%), cortical III layer (37.8%), CA4 (35.3%), cortical V layer (34.1%), cerebellum (28.2%), CA3 (24.3%), CA2 (16.2%), and CA1 (2.0%). Only the CA1 showed an almost total neuronal loss. In contrast, a single postictal injection of CA-074 or E-64c led to significant inhibition of postischemic neuronal death in all brain regions studied. Overall, more surviving neurons were seen after E-64c treatment than with CA-074: cerebellum, 91.6% vs 85.6%; CA4, 88.6% vs 77.3%; caudate nucleus, 86.1% vs 89.8%; CA2, 83.6% vs 53.0%; outer putamen, 81.3% vs 87.7%; CA1, 80.1% vs 47.4%; CA3, 79.6% vs 60.3%; cortical layer III, 75.5% vs 67.7%; and cortical layer V, 75.0% vs 65.9%, for E-64c and CA-074, respectively. Cathepsin plays a critical role in ischemic neuronal death, and its inhibitors may protect neurons throughout the brain.     

MRI findings on healing process of vertebral fracture in osteoporosis

The healing process of vertebral fracture was investigated in 37 senile osteoporotic patients on serial magnetic resonance imagings (MRIs), including enhancement study with gadolinium-diethylenetriaminepentaacetic acid (Gd-DTPA). The healing process was classified retrospectively into six categories in terms of the changes and recovery of T1 and T2 signal intensity and in terms of the local vascularity. Two types of damage foci were identified, and, in each type, three patterns of healing were noted. In the partial collapse type, the focus of damage in the fractured vertebral body was located near the cranial or caudal endplates; in the total collapse type, the focus was located at the center of the body. In each of these two types, fracture healing was smooth, belated, or resulted in nonunion. In the partial collapse type, vertebral fractures healed smoothly (smooth pattern) in 8 cases, belatedly (belated pattern) in 11, and resulted in nonunion (nonunion pattern) in 5. In the total collapse type, vertebral fractures healed smoothly (smooth pattern) in 6 cases, belatedly (belated pattern) in 4, and resulted in nonunion (nonunion pattern) in 3. The percent height of the anterior wall, central portion, and posterior wall was defined to allow accurate calculatation of the collapse of the fractured vertebral body. Chronological changes in the vertebral body height were investigated. Progressive collapse of the vertebral body was minimal in the smooth pattern cases, and most severe in the nonunion pattern cases. Statistical analysis indicated that prediction of the course of the fracture was difficult only with the height of the fractured vertebral body in acute phase. Enhancement study with Gd-DTPA showed that, in fractures with favorable prognosis, the ischemic area in the body tended to be smaller from the beginning, and restoration of vascularity was prompt. On the contrary, in fractures with unfavorable prognosis, the ischemic area was wider, and restoration of vascularity was poor.     

Clinical manifestation of small thalamic hemorrhage

CT scan is useful for the simultaneous evaluation of the relation between the thalamic lesions and the clinical manifestations. According to CT findings, twenty-three patients with thalamic hemorrhage measuring less than 2 cm in size could be classified into 4 groups: 1) anterior group--hematoma located in the anterior nuclear group, 2) medial group--hematoma located in the medial nuclear group, 3) lateral group--hematoma located in the lateral nuclear group close to the internal capsule, 4) posterior group--hematoma located in the pulvinar. The clinical manifestations of both the anterior and medial groups were characterized by the disturbance of consciousness followed by the mental impairment; the lateral group, by the hemiparesis or hemiplegia with the sensory disturbance, and the posterior group, especially with left thalamic lesions, by the speech disturbance. The motor palsy in cases of thalamic hemorrhage differed from that of putaminal hemorrhage: the patients with thalamic hemorrhage could move their fingers despite being unable to move their shoulders and elbows, or the motor weakness was more severe in their lower extremities than in their upper ones. As the sensory disturbance, the sensory impairment (hypesthesia) was frequently associated with the numbness (dysesthesia). The prognosis of motor palsy, ocular manifestations, and speech disturbance was good, whereas that of sensory and mental disturbance was not always good.