Evaluation of in vivo and in vitro repopulation assays for murine haemopoietic stem cells

Three assays to determine the repopulation potential of stem cells in murine bone marrow grafts were evaluated on their reliability with regard to the ranges in graft size. In vivo, marrow repopulating ability, as calculated from the number of in vitro clonable progenitors (colony-forming units in culture, CFU-C) generated by the graft in the femur of an irradiated recipient, appeared to be independent of the input over a wide range of cell numbers grafted. A second assay, erythroid repopulating ability is a measure of the number of new reticulocytes or erythrocytes in the blood generated by the graft, and significantly underestimates stem cell activity of the graft. The third assay measures the long-term repopulating ability of bone marrow cells in vitro on pre-established stromal cell layers by determination of the number of CFU-C produced in these cultures. Calculations of short-term in vitro repopulating ability, done from measurements of the production of non-adherent CFU-C in the first week of culture, appeared to be independent of cell input. Long-term in vitro repopulating potential, measured by the CFU-C content of the adherent layer at 4 weeks, is also independent of the numbers of cell input.     

Accuracy and choice of procedures in 24-hour oesophageal pH monitoring with monocrystalline antimony electrodes

In 24 h pH monitoring, the evaluation is dependent on the absolute accuracy of the pH measurements. Several sources of error exist, such as the chemical composition of calibration buffers and reference electrode gel and the effect of temperature on both the pH and the reference electrodes. We investigated the magnitude of these errors for the monocrystalline antimony electrode. Similar analysis applies to other types of pH electrodes. The errors we found are important when choosing a calibration procedure. We recommend a calibration procedure in which the pH and reference electrodes are both put in a beaker with the calibration buffers prior to and after the 24 h measurements. The calibration buffers and the electrode gel should have a specially selected ion composition where, for example, the Cl-ion concentration is critical. Corrections for differences in temperature between the calibration and the in situ measurements must be added. The pH measurements can be checked by performing in situ calibration.     

Predominance of methicillin-resistant Staphylococcus aureus in the residents and environments of long-term care facilities in Taiwan

Background/purpose

This study investigated the distribution and persistence of multidrug resistant organisms (MDROs) including methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Enterobacteriaceae (CRE), carbapenem-resistant Pseudomonas aeruginosa (CRPA), and multidrug-resistant Acinetobacter baumannii (MDRAB) in six long-term care facilities (LTCFs).

PKA参与脊髓背角C-纤维诱发电位LTP的诱导和早期维持

目的: 探讨环一磷酸腺苷依赖的蛋白激酶 (PKA) 在脊髓背角C-纤维诱发电位长时程增强(LTP)的诱导和维持中的作用。方法:细胞外记录技术在脊髓腰膨大部记录背角浅层神经元C-纤维诱发电位。 结果:(1) 8-Br-cAMP诱发脊髓背角C-纤维诱发电位LTP,且8-Br-cAMP-诱导的 LTP 遮蔽强直刺激诱导的LTP。(2) PKA的选择性抑制剂Rp-CPT-cAMPS阻断C-纤维诱发电位LTP的诱导和时间依赖性翻转C-纤维诱发电位LTP。(3)蛋白质合成抑制剂茴香霉素彻底阻断8-Br-cAMP诱发的 LTP。(4)MAPK选择性抑制剂PD98059阻断8-Br-cAMP诱发的LTP。 结论:脊髓背角神经元存在PKA信号途径,并参与脊髓背角C-纤维诱发LTP的诱导和早期维持。     

Long-term potentiation in the interpositus and vestibular nuclei in the rat

Summary Previous unpublished experiments from this laboratory had revealed only post-activation depression effects in the cerebellar cortex when its inputs were activated by high frequency trains. In the experiments reported in this paper, we found reliable long-term potentiation (LTP) effects in the deep nuclei (interpositus and vestibular) when stimulation trains were applied to the white matter at the point where inferior peduncle fibers enter the cerebellum. LTP effects were found in both acute and chronic preparations. In the chronic preparations, LTP lasted for at least 8 days in all but one animal.     

Postextrasystolic potentiation does not distinguish ischaemic from stunned myocardium

Myocardial function is impaired by ischaemia, and it remains depressed during reperfusion following short periods of ischaemia (stunned myocardium). We tested whether ischaemic and reperfusion dysfunction, in particular the time course of its recovery, can be distinguished by postextrasystolic potentiation (PESP). In eight open-chest dogs, posterior systolic wall thickening (sonomicrometry) was reduced by graded occlusion of the left circumflex coronary artery (LCX) from 17.4±6.8% (SD) during control conditions to 10.7±1.3% (mild ischaemic dysfunction), 7.2±2.3% (moderate ischaemic dysfunction), 3.6±1.4% (severe ischaemic dysfunction), and -4.4±3.6% (complete coronary occlusion). Extrasystoles with constant prematurity and a fully compensated postextrasystolic interval were induced after at least 4 min steady-state ischaemia. After each ischaemic period full recovery of posterior systolic wall thickening was assured. During 8 h of reperfusion following a 15-min LCX occlusion, extrasystoles were induced when posterior systolic wall thickening was comparable to one degree of the preceding ischaemic dysfunction. The increases in posterior systolic wall thickening induced by PESP were 10.5±5.8% during control conditions, during ischaemia they were 11.5±3.5% (mild dysfunction), 12.3±4.6% (moderate dysfunction), 12.6±4.1% (severe dysfunction) and 10.4±4.4% (complete coronary occlusion), and during reperfusion they were 12.8±8.2% (severe dysfunction), 13.0±9.7% (moderate dysfunction) and 10.7±2.2% (mild dysfunction). These increments in systolic wall thickening as well as those in ejection thickening were not significantly different. PESP can thus not distinguish between ischaemic and reperfusion dysfunction nor between different degrees of myocardial dysfunction.This study was supported by the Deutsche Forschungsge-meinschaft (He 1320/3-2). cand. med. S. Schäfer was involved in some of these experiments and presented part of the data at the 56th Annual Meeting of the Deutsche Gesellschaft für Herz- und Kreislaufforschung in Mannheim (Z Kardiol 79 [Suppl 1]: 24,1990). Part of the data were also presented at the 11th Congress of the European Society of Cardiology in Nice (Eur Heart J 10 [Suppl]: 242, 1989) and at the 73rd Annual Meeting of the Federation of American Societies for Experimental Biology in New Orleans (FASEB J 3: A841, 1989)     

Persistent enhancement of transmitter release accompanying long-term potentiation in the guinea pig hippocampus

In order to examine temporal changes in enhancement of transmitter release during long-term potentiation (LTP), we examined amplitude fluctuation of excitatory postsynaptic potentials (EPSPs) for longer periods than 2 h after tetanic stimulation (up to 4 h in the longest observation). The relative magnitude of excitatory postsynaptic potentiation (EPSP) fluctuation (coefficient of variation, CV) reduced throughout the observation periods in association with an increase in EPSP amplitude after tetanic stimulation. The reciprocals of squared CVs (= mean²/variance) were almost in proportion to the magnitude of LTP, and the ratio of 1/CV² and the LTP magnitude did not change significantly for up to 4 h. These findings suggest that a prolonged enhancement of transmitter release from presynaptic terminals underlies LTP, and the relative contribution of this presynaptic enhancement does not change significantly for 2 h (maybe up to 4 h, or longer) after tetanic stimulation.     

Reduction of ECS produced retrograde amnesia by post-trial introduction of strychnine

Strychnine sulfate was used to reduce the amount of retrograde amnesia induced by electroconvulsive shock (ECS). Male Swiss-Webster mice were trained on the step-down passive avoidance task, given ECS and then injected intraperitoneally with either 0.1 mg/kg strychnine or saline. The animals were tested in the apparatus one to twenty-four hours later. Mice given ECS alone displayed partial memory for the training (by responding with longer latencies) if tested within two hours after training, but did not display retention twenty-four hours later. Animals given strychnine following ECS had significantly longer step-down latencies at twenty-four hours than did the saline or no injection controls. Further experiments replicated the basic results, and also showed that strychnine injections delayed two hours after the training trial attenuated ECS-induced retrograde amnesia, but that a three hour injection delay was ineffective. The results were interpreted in terms of changes in short-term and long-term memory.     

The Effects of Corticoliberin at Different Concentrations on Long-Term Potentiation in Slices of Rat Olfactory Cortex

Experiments were performed to investigate evoked focal potentials in slices of rat olfactory cortex. The results showed that 1 M corticotropin-releasing factor (CRF; corticoliberin) increases the frequency of posttetanic potentiation, which had a shorter delay phase than in control experiments. Posttetanic potentiation during perfusion with 0.1 M corticoliberin had a longer delay phase than in control experiments.