An attempt was made to clarify the effects of the modified NLA on the central nervous system using Diazepam and Pentazocine. The unanesthetized rabbit was used as the experimental animal. The results obtained were as follows. 1) Threshold of arousal reaction and evok ed muscular discharge rose more remarkably when Diazepam and Pentazocine were administered in the ratio of 1 to 3 than when either one of them was administered. This finding showed that the combination of the two agents had more inhibitory effect on consciousness and motor activity than either of them. 2) Average threshold of arousal reaction in the hippocampus rose more rem arkably than in the cerebral cortex when the two agents were administered in the same way. 3) Under the combination of the two agents, average threshold rose most remarkably in hind limbs, followed by fore limbs, the hippocampus and the cerebral cortex in this order. 4) Under the combination of the two agents, threshold of evoked muscular d ischarge showed a marked elevation, about double the threshold of arousal reaction. This finding proved that the modified NLA had an inhibitory effect on the sensory nerve, and an even more powerful inhibitory effect on the motor nerve. 5) Pentazocine showed as equally inhibitory effect as the combination of the two agents did on the early components of average evoked potential in the cerebral cortex. Consequently, it was clear that the effect on the afferent pathway was mostly attributed to Pentazocine. 6) The late components (N4 and N5) of the average evoked potential in creased in the cerebral cortex under the combination of the two agents. 7) The late components of the average evoked potential in the hippocampus were powerfully inhibited under the same condition. This finding made it clear that the combination of the two agents powerfully suppressed pain reaction. 8) M and H waves showed no incre ase under the combination of the two agents, whereas they did with the administration of either one of them. This fact is advantageous to anesthetization in a surgical operation. 9) The combina tion of the two agents showed powerful inhibition on the microvibration (MV). 10) With regard to photopalpebral reflex (PPR), PPR6 showed an increase under the combination of the two agents. 11) The rate of in crease of the threshold of arousal reaction and evoked muscular discharge diminished remarkably after the destruction of nucleus ventralis anterior (VA) under the combination of the two agents. This finding elucidated that the VA was concerned with the increase of the threshold of arousal reaction and evoked muscular discharge. 12) The increase of N4, N5 and PPR6 under the combination of the two agents disappeared after the destruction of VA. It was therefore obvious that the increase of these responses was caused by excitement of the diffuse thalamocortical projection system. 13) Recruiting response caused by excitement of the d iffuse thalamocortical projection system showed an increase under the combination of the two agents. 14) The action potentials, VAN4 and VAN5, induced from the cerebral cortex by a 1Hz stimulation on the VA increased under the combination of the two agents. This finding led us to the conclusion that the increase in N4, N5 and PPR6 was a direct expression of the excitement of the diffuse thalamocortical projection system.
To date, a number of resarchers have reported that platelets contain some growth factors, including Platelet-derived growth factor (PDGF), Epidermal growth factor (EGF), Transforming growth factor-β (TGF-β) and others. These growth factors are considered to play a role in restoration of wounded blood vessels, arteriosclerosis, metastasis of malignant tumors and other vascular diseases. Though each growth factor has been investigated in details, the study of interaction of these growth factors in vivo has been neglected. The author investigates, by the use of flow cytometry, how these growth factors as a whole stimulate the proliferation of fibroblasts. These recent studies found that a part of those growth factors are concerned in the prrogress of the G0→G1 phase and another part is concerned in the progress of the G1→S phase. The former growth factor is called the “competence factor”, and the latter is called the “progression factor”. The studies to date suggest that PDGF acts as “competence factor” in the cell growth and that EGF and TGF-β; act as “progression factors”. The author's study suggests that the platelet-dependent growth factors, as a whole, act as “competence factors” on 3T3 fibroblasts, but do not act on SV40 transformed 3T3 fibroblasts. On the other hand, this study points out that action as the “progression factor” of those growth factors is not a majar one in the proliferation of both fibroblasts. The author surmises that platelet-dependent growth factors in cooperation turn the cells of blood vessel from G0 phase (passive phase) to G1 phase (activate phase) and couse proliferation of the cells of blood vessels in cooperation with other serum factors, when the blood vessels are damaged.
Neutrophil chemotaxis is an important mechanism in the first stage of the inflammatory response against the invasion of microorganism in human and other species. During the past 100 years many investigations have looked at the fundamentals of neutrophil chemotaxis in vitro and went on to develop many types of assay to quantify neutrophil chemotaxis in the laboratory. In 1962, Boyden reported a new method of neutrophil chemotaxis with a special chamber with a lower compartment, where a chemoattractant was placed separated by a filter on top of which a cell suspension was placed. In searching for a sensitive and more conv inient method of neutrophil chemotaxis, we have modified the previous method of 51-chromium labeled neutrophil chemotaxis. Normal human subjects, aged 24 to 37 years, donates the blood. Polymorphonuclear cells were purified by using Ficoll-Hypaque and dextran sedimentation. Final chromium uptake was determined to be 2.78±0.98 % of the original count. We used acrylic blind well chemotactic chambers which were divided using 3 or 5, um pore sized polycarbonate filter (Nuclepore) (13mm diameter) on top of 3 or 5 μm pore sized cellulose ester filter (Millipore)(13mm diameter) and compared to original method. To find the optimum incubation time for chemotaxis using a 5 μm Nuclepore on top of the 5 μm Millipore, we counted all of the compartments of this chamber system. By varying incubation time from 15 to 150 minutes, we noted that the neutrophils begin migration within 15 minutes. At 60 minutes, a marked difference was detected in the lower filters of the chambes with EAP vs media. As incubation time was lengthened mere counts were found in the lower filters and less counts were found in the upper chamber fluids. Chemotactic activity was correlated with the cell concentration from 2.875×105 cells/ml to 6.9×106 cells/ml. In our 22 experiments using 5, μm Nuclepore filters with 5 pm Millipore filters, the target: non target ratio was 4.68±1.83when incubated for 90min. These results sho w that our method using Nuclepore on top of a Millipore filter reduced incubation time from 3 hours to 90 minutes while maintaining successful chemotaxis. A shorter incubation time maximizes laboratory efficiency by reducing spontaneous 51-chromium release from injured and dying cells yielding more reproducible results and also by allowing a laboratory to test more samples in less time.