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.     

Calcium intake in the elderly: validation of a dietary questionnaire

Epidemiological studies of calcium and osteoporosis have been hampered by the lack of a suitable tool for assessing calcium intake. This report describes a new frequency and amount questionnaire for measuring present and past calcium intake in the elderly. The validity of the questionnaire was tested against two commonly used standards of dietary assessment, five-day duplicate diets and seven-day weighed dietary inventories. The resulting correlation coefficients were, respectively, r = 0.76 and r = 0.69, while that for repeatability was r = 0.84. Furthermore, the questionnaire categorized subjects into thirds of the distribution of intake with almost no gross misclassification. It is suggested that the present findings may be extended to the majority of normal, healthy elderly subjects, implying wide application for the questionnaire in the assessment of calcium intake in the elderly.     

Insulinoma: Selective arterial calcium injection performed to confirm localization and complete resection

A case of insulinoma is reported in a patient in whom selective arterial calcium injection (SACI) tests were performed both to confirm tumor localization before surgery and to confirm complete tumor removal during surgery. An 18-year-old woman with hypoglycemic episodes was diagnosed with an insulinoma in the pancreatic body demonstrated by celiac arteriography. In a preoperative SACI test, calcium was injected into the splenic artery (SpA), gastroduodenal artery (GDA), and superior mesenteric artery (SMA). Serum immunoreactive insulin (IRI) and proinsulin levels were measured in hepatic venous samples. IRI was markedly increased after the injection of calcium into the GDA and SMA, while there was no response in IRI levels when calcium was injected into the SpA. Therefore, no occult insulinoma was revealed in the distal area fed by the SpA, although the presence of insulinoma was uncertain in the proximal pancreas. In the intraoperative SACI test, calcium was injected into the celiac artery. Insulin (determined by enzyme immunoassay) and proinsulin levels were measured in portal venous samples before and after resection of the tumor. After resection, these levels decreased in response to the calcium stimuli, confirming complete removal of the insulinoma. The SACI test was helpful to localize the insulinoma and was useful to confirm the complete removal of the tumor.     

0～16岁城乡儿童骨骼矿物质含量及影响因素的研究

使用单光子骨矿物测定仪，自１９９４年初到１９９５年底，分别在河南省扶沟县和新乡市测定了３３２名健康的０～１６岁城乡儿童右前臂１／３处桡骨和尺骨的骨矿物质含量（ＢＭＣ），采用逐步多元回归分析了多个独立变量对桡骨ＢＭＣ和测量部位骨宽度的影响。结果显示：桡骨和尺骨的ＢＭＣ、骨宽度（ＢＷ）、ＢＭＣ／ＢＷ和前臂长度均随年龄而增加，０～６岁城市儿童的ＢＭＣ高于农村儿童；桡骨ＢＭＣ与所测量部位的ＢＷ呈正相关，年龄、体重和身高对桡骨ＢＭＣ都显示出显著和独立的正影响；儿童的桡骨ＢＭＣ与年龄呈显著正相关；而桡骨ＢＭＣ与体重的比值与年龄呈显著负相关；测定部位桡骨ＢＷ和前臂长度随年龄的增加呈曲线增加。在以身高和体重调整后，性别、年龄和体重对桡骨ＢＷ的影响变得不显著，只有身高的影响显著。本研究证明儿童在０～１６岁范围，身高（而不是肥胖）是骨骼ＢＭＣ的主要决定因素。在比较儿童骨密度结果时，要参考多个变量，如ＢＭＣ、骨密度，ＢＭＣ／体重、ＢＭＣ／年龄等综合评价指标更有说明力。     

Focus groups: their role in developing calcium-related education materials

Because attitudes concerning a topic can diminish the effectiveness of educational materials, previously identified attitudes concerning calcium intake were explored through focus group interviews during the developmental stages of calcium education materials. Although four focus groups of six to seven participants were planned, each of the four groups consisted of two to six women. All focus groups followed the same format, lasting for 60–90 min; questions progressed from the general to more specific. The focus groups revealed several attitudinal barriers toward dietary behavioural change, including lack of prior interest in the topic and lack of time. Attitudes about dairy calcium included the belief that dairy foods were high in fat and should be avoided, and the belief that dairy foods would cause stomach upsets. Also, neither younger nor older women felt that osteoporosis was a problem their age group needed to address. Readability scales were not necessarily predictive of preference. This study shows that focus group interviews make a valuable contribution to planning and evaluating nutrition education materials.     

Study on lymphocyte activation and proliferation induced by anti-CD3 McAb

Summary T cell activation and proliferation via CD₃-TCR complex were investigated by lymphocyte DNA synthesis in vitro. Several interfering factors were also discussed. The result indicated that lymphocyte activation and proliferation are calciumdependent. A rise of cytoplasmic free Ca²⁺ quickly following activation with CD₃ McAb is mainly due to intracellular mobilization of Ca²⁺, while lymphocyte proliferation needs both intracellular mobilization of Ca²⁺ as well as influx of extracellular Ca²⁺. It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte³H-TdR incorporation obviously. PLC and PKC inhibitor neomycin and P. S. S could also decrease T cell proliferation.     

离体灌流鼠心胞浆游离钙——fura—2测定

本实验建立了一种浮动基点式ｆｕｒａ－２测定离体灌流心脏细胞内游离钙浓度的新模型，本模型中游动基点式测定地克服目前ｆｕｒａ－２技术中受细胞内某些物质产生的自身荧光变化干扰的缺点，实现了自动消除细胞自身荧光变化对实验结果的影响，并在此基础上将广泛用于测定游离状态的细胞内游离钙的ｔｕｒａ－２技术推广到离体灌流心脏，实现了在更接近心肌生理状态的条件下测定其胞浆游离钙。     

Involvement of glutamate receptors,lipases, and phospholipases in long-term potentiation and neurodegeneration

Glutamate is the primary excitatory amino acid in the mammalian central nervous system. Normal excitation of glutamate receptors initiates the stimulation of phospholipases and lipases with the generation of second messengers that are necessary for normal cell function. The overstimulation of glutamate receptors can initiate a cascade of biochemical events including stimulation of membrane phospholipid turnover, excessive calcium entry, abnormal phosphorylation, and proteolysis. These events may be responsible for neuronal injury and degeneration found in Alzheimer disease, ischemia, spinal cord trauma, epilepsy, and Huntington disease.     

用Fura-2测定周围血单个核细胞内游离钙浓度

本文用Fura-2测定了周围血单个核细胞内游离钙浓度，并对有关实验条件进行了探讨。对比研究表明，血标本在4℃保存4小时对测定结果无显著影响，测定标本中单个核细胞数以10~5～10~6/ml为宜。认为测定单个核细胞内游离钙浓度对判断单个核细胞的功能状况有一定帮助。