Ji Hyun Kim;Gen Murakami;Jose Francisco Rodriguez-Vazquez;Ryo Sekiya;Tianyi Yang;Sin-ichi Abe
Anatomy and Cell Biology
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v.57
no.2
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pp.278-287
/
2024
Striated muscle insertions into the skin and mucosa are present in the head, neck, and pelvic floor. We reexamined the histology of these tissues to elucidate their role in transmission of the force. We examined histological sections of 25 human fetuses (gestational ages of ~11-19 weeks and ~26-40 weeks) and 6 cadavers of elderly individuals. Facial muscle insertion or terminal almost always formed as an interdigitation with another muscle or as a circular arrangement in which muscle fiber insertions were sandwiched and mechanically supported by other muscle fibers (like an in-series muscle). Our examination of the face revealed some limited exceptions in which muscle fibers that approached the dermis were always in the nasalis and mentalis muscles, and often in the levator labii superioris alaeque nasi muscle. The buccinator muscle was consistently inserted into the basement membrane of the oral mucosa. Parts of the uvulae muscle in the soft palate and of the intrinsic vertical muscle of the tongue were likely to direct toward the mucosa. In contrast, the pelvic floor did not contain striated muscle fibers that were directed toward the skin or mucosa. Although 'cutaneous muscle' is a common term, the actual insertion of a muscle into the skin or mucosa seemed to be very rare. Instead, superficial muscle insertion often consisted of interdigitated muscle bundles that had different functional vectors. In this case, the terminal of one muscle bundle was sandwiched and fixed mechanically by other bundles.
Objectives : This is the experimental paper to study the curative and preventive effects of Dansambohyultang, Methods : Oriental herb medicine for gastro-intestinal disease caused by stress, on the rats put under the stress through starvation, heating and immoblization, the author used four different group; the normal group, which was not put under stress and well fed, and the control group, to which the drug was not administered and put under the stress through starvation, heating and immoblization, the Sample I, to which the drug was administered before they were put under stress, the Sample lI, to which the drug was administered after they were put under stress. Results : When a Masson's trichrome stain method was applied to the control group, a small size of ulcer was found in the fundus and the pylorus, and atrophy was observed in the neck region of mucous membrane. When the drug was administered to the Sample I and the Sample II, the former recovered from the ulcer and atrophy to almost normal and showed better results than the latter. When a Masson's trichrome stain method was applied to the control group, atrophy of mucous membrane was found all over the intestines. When the drug was administered to the Sample I and the Sample ll, the curative and recovered effects was seen in both, but the former showed better results than the latter. After application of Mayer's hematoxylin stain method, the observation of numerical changes of goblet cells in the small intestines showed the most significant decrease in the control group. In the Sample I, the number of the cells in the duodenum, the jejunum and the ileum returned to an almost normal level. In the Sample II, a slight increase was observed only in the ileum. After application of Mayer's hematoxylin stain method, the observation of numerical changes of goblet cell in the large intestines showed a significant decreased in the control group. Both the Sample I and the Sample II showed an increase in the number of goblet cells in comparison with the control group. When a serotonin-immunoreaction test was applied, the control group showed a general decrease in the number of gastro-entero-endocrine cells, The Sample I showed the effect in all over gastro.intestinal tract as compared with control group and the Sample II showed the effect in all over gastro-intestinal tract except the duodenum and the former showed better results than the latter. Conclusion : According to the above results, the regions which are most sensitive to stress were observed to the ileum and the colon. It was concluded that Dansambohyultang has a remedical value particularly on the abnormalities caused by stress in all over gastro-intestinal tract and it was found to be more effective in the Sample I than in the Sample ll. Judging from the fact that the Sample I showed better results than the Sample ll, Dansambohyultang can be said to have a preventive effect more than a curative effect on gastro-intestinal tract disease.
Hong, Young Mi;Yoon, Woong Hee;Lee, You Ra;Kim, Soo Ji;Ngabire, Daniel;Narayanasamy, Badrinath;Ornella, Mefotse Saha Cyrelle;Kim, Myunghee;Cho, Euna;Lee, Bora;Hwang, Tae-Ho
Journal of Life Science
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v.31
no.11
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pp.1028-1036
/
2021
Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.
Chloride($Cl^-$) channels play critical roles in cell homeostasis and its specific functions such as volume regulation, differentiation, secretion, and membrane stabilization. The presence of these channels have been reported in all kinds of cells and even in frog oocytes. These essential role of $Cl^-$ channels in cell homeostasis possibly play any role in egg homeostasis and in the early stage of development, however, there has been no report about the presence of $Cl^-$ channel in the mammalian oocyte. This study was performed to elucidate the presence of $Cl^-$ channels in hamster eggs. When allowing only $Cl^-$ to pass through the channel of the egg membrane by using impermeant cation such as N-methyl-D-glucamine(NMDG), single channel currents were recorded. These channel currents showed typical long-lasted openings interrupted by rapid flickering. Mean open $time({\tau}o)$ was 43${\pm}$10.14 ms(n=9, at 50 mV). The open probability(Po) was decrease with depolarization. The current-voltage relation showed outward rectification. Outward slop conductance(32${\pm}$5.4 pS, n=22) was steeper than the inward slop conductance(10${\pm}$1.3 pS). Under the condition of symmetrical 140 mM NaCl, single channel currents were reversed at 0 mV(n=4). This reversal potential(Erev) was shifted from 0 mV at 140 mM concentration of internal NaCl(140 mM [Na+]i) to 9.8${\pm}$0.5 mV(n=4) at 70 mM [Na+]i and 11.5${\pm}$1.9 mV at 280 mM [Na+]i(n=4) respectively, strongly suggesting that these are single $Cl^-$ channel currents. To examine further whether this channel has pharmacological property of the $Cl^-$ channel, specific Cl channel blockers, IAA-94(Indanyloxyacetic acid-94) and DIDS(4, 4'-diisothiocyan ostillben- 2-2'disulfonic acid) were applied. IAA-94 inhibited the channel current in a dose-dependent manner and revealed a rapid and flickering block. From these electrophysiological and pharmacological resluts, we found the novel $Cl^-$ channel present in the hamster oocyte membrane. The first identification of $Cl^-$ channel in the hamster oocyte may give a clue for the further study on the function of $Cl^-$ channel in the fertilization and cell differentiation.
Background: Among the injurious agents to which the lung airspaces are constantly exposed are reactive species of oxygen. It has been widely believed that reactive oxygen species may be implicated in the etiology of lung injuries. In order to elucidated how this oxidant causes lung cell injury, we investigated the effects of exogenous $H_2O_2$ on alveolar epithelial barrier characteristics. Methods: Rat type II alveolar epithelial cells were plated onto tissue culture-treated polycarbonate membrane filters. The resulting confluent monolayers on days 3 and 4 were mounted in a modified Ussing chamber and bathed on both sides with HEPES-buffered Ringer solution. The changes in short-circuit current (Isc) and monolayer resistance (R) in response to the exogenous hydroperoxide were measured. To determine the degree of cellular catalase participation in protection against $H_2O_2$ injury to the barrier, experiments were repeated in the presence of 20 mM aminotriazole (ATAZ, an inhibitor of catalase) in the same bathing fluid as the hydroperoxide. Results: These monolayers have a high transepithelial resistance (>2000 ohm-$cm^2$) and actively transport $Na^+$ from apical fluid. $H_2O_2$(0-100 mM) was then delivered to either apical or basolateral fluid. Resulting indicated that $H_2O_2$ decreased Isc and R gradually in dose-dependent manner. The effective concentration of apical $H_2O_2$ at which Isc (or R) was decreased by 50% at one hour ($ED_{50}$) was about 4 mM. However, basolateral $H_2O_2$ exposure led to $ED_{50}$ for Isc (and R) of about 0.04 mM. Inhibition of cellular catalase yielded $ED_{50}$ for Isc (and R) of about 0.4 mM when $H_2O_2$ was given apically, while $ED_{50}$ for basolateral exposure to $H_2O_2$ did not change in the presence of ATAZ. The rate of $H_2O_2$ consumption in apical and basolateral bathing fluids was the same, while cellualr catalase activity rose gradually with time in culture. Conclusion: Our data suggest that basolateral $H_2O_2$ may affect directly membrane component (e.g., $Na^+,\;K^+$-ATPase) located on the basolateral cell surface. Apical $H_2O_2$, on the other hand, may be largely degraded by catalase as it passes through the cells before reaching these membrane components. We conclude that alveolar epithelial barrier integrity as measured by Isc and R are compromised by $H_2O_2$ being relatively sensitive to basolateral (and insensitive to apical) $H_2O_2$.
Background : The cell-mediated immune response plays an important role in tuberculosis. After being activated by mycobacterial antigens, T lymphocytes express a high affinity receptor (IL-2R) for interleukin-2 (IL-2) on their own surface and release a soluble fraction of the IL-2 receptor (sIL-2R) from the cell membrane into the circulation. Neopterin is a metabolite of guanosine-triphosphate, which is produced by stimulated macrophages under the influence of IFN-$\gamma$ with a T lymphocyte origin. Therefore, the utility of sIL-2R, IFN-$\gamma$ and the neopterin levels as immunologic indices of the cell-mediated immune response and severity of disease in patients with pulmonary tuberculosis was assessed. Methods : The serum sIL-2R, IFN-$\gamma$ and neopterin levels were measured in 39 patients with pulmonary tuberculosis, 6 patients with tuberculous lymphadenitis prior to treatment and 10 healthy subjects. The serum and pleural sIL-2R, neopterin and ADA levels were measured in 22 patients with tuberculous pleurisy. The patients with pulmonary tuberculosis were divided into a mild, moderate and severe group according to the severity by ATS guidelines. To compare the results from these patients with those of the pretreatment levels, the sIL-2R, IFN-$\gamma$ and neopterin levels were measured in 36 of the 39 patients(1 patient, expired; 2 patients were referred to a sanitarium) with pulmonary tuberculosis after 2 months of treatment. Results : 1) the serum sIL-2R and IFN-$\gamma$ levels were elevated in patients with tuberculosis when compared to those of healthy subjects (p>0.05). The neopterin concentration in the serum was significantly lower in patients with pulmonary tuberculosis($2967{\pm}2132.8$ pg/ml) than in healthy controls($4949{\pm}1242.1$ pg/ml)(p<0.05). 2) In the pulmonary tuberculosis group, the serum sIL-2R and IFN-$\gamma$ levels were higher in patients with severe disease than those in patients with mild and moderate disease. However, the neopterin levels declined as the pulmonary tuberculosis became more severe (p<0.01). 3) The mean serum sIL-2R and IFN-$\gamma$ levels declined from $1071{\pm}1139.4$ U/ml to $1023{\pm}1920.9$ U/ml(p>0.05), $41{\pm}52.8$ pg/ml to $22{\pm}23.9$ gm/ml(p<0.05), respectively, after 2 month of treatment. The mean serum neopterin levels increased from $3158{\pm}2272.6$ pg/ml to $3737{\pm}2307.5$ pg/ml(p>0.05) after a 2 month of treatment. These findings were remarkable in the severe group of pulmonary tuberculosis with a clinical correlation. 4) In the patients with tuberculous pleurisy, the serum sIL-2R and ADA were significantly higher than those in the pleural fluid, However, the neopterin levels in the sera and pleural effusion were similar. Conclusion : On the basis of this study, sIL-2R, IFN-$\gamma$ and neopterin measurements may not only provide an insight into the present state of the cell-mediated immune response, but also serve as parameters monitoring of the prognosis of the disease, particularly in patients with severe pulmonary tuberculosis. In addition, an assay of the pleural sIL-2R levels might signal a stimulated local immunity including T cell activation in the tuberculous pleural effusion.
Effect of a $Na^+$ gradient on $Ca^{2+}$ uptake was studied in isolated sarcolemmal vesicles of cat ileal longitudinal muscle. $Ca^{2+}$ uptake was markedly stimulated in the presence of an outwardly directed $Na^+$ gradient. External $Na^+$, monensin and A23187 abolished the $Na^+-dependent$$Ca^{2+}$ uptake. Monovalent cations such as $K^+$, $Li^+$, $Rb^+$, $Cs^+$ and choline could not substitute for $Na^+$ in enhancement of $Ca^{2+}$ uptake. Divalent cations such as $Ba^{2+}$, $Sr^{2+}$, $Mn^{2+}$ and $Cd^{2+}$ but not $Mg^{2+}$ inhibited the $Na^+-dependent$$Ca^{2+}$ uptake. Increase in external pH in the range of 6.0 to 8.0 stimulated the $Na^+-dependent$$Ca^{2+}$ uptake. Amiloride inhibited the $Na^+-dependent$$Ca^{2+}$ uptake at concentrations above 0.5 mM, whereas diltiazem or vanadate did not. The apparent Km of the $Na^+-dependent$$Ca^{2+}$ uptake for $Ca^{2+}$ was 18.2 ${\mu}M$ and apparent Vmax was 689.7 pmole/mg protein/5 sec. Kinetic analysis of the $Na^+-dependent$$Ca^{2+}$ uptake showed a noncompetitive interaction between internal $Na^+$ and external $Ca^{2+}$. The dependence of $Ca^{2+}$ uptake on internal $Na^+$ showed sigmoidal kinetics and Hill coefficient for internal $Na^+$ was 2.52. Inside positive membrane potential generated by imposing an inwardly directed $K^+$ gradient and valinomycin significantly stimulated the $Na^+-dependent$$Ca^{2+}$ uptake. These results indicate that a $Na^+-Ca^{2+}$ exchange system exists in the sarcolemmal membranes isolated from cat ileal longitudinal muscle and it might operate as an electrogenic process.
The characteristics of anion exchange with internal $HCO_3\;^{-}\;(or\;OH^-)$ was studied by determining the time course of hemolysis in isoosmotic ammonium salt solution in human erythrocytes. The effects of inhibitors, pH and temperature on the exchange between internal $HCO_3\;^-\;(or\;OH^-)$ and external $Cl^-$ were observed and the permeabilities of various organic and inorganic anions were also measured. The results were compared with data previously reported from the experiments using radioisotopes. The results are as follows; 1) SITS $H_2DIDS$ and furosemide inhibited the hemolysis of erythrocytes in isoosmotic $NH_4Cl$ solution in a dose·dependent manner, and the concentrations for lengthening twice the time for $half-hemloysis(t_{1/2})\;were\;2.3{\times}10^{-7},\;1.3{\times}10^{-7}\;and\;2.5{\times}10^{-5}M$, respectively. 2) Acetazolamide also shifted the time-dependent hemolytic curve to the right in a dose-dependent manner, and the concentrations for lengthening twice $t_{1/2}\;was\;2.4{\times}10^{-5}M$. 3) The time-dependent hemolysis was delayed by decreasing pH from 7.0 to 6.2, but w·as not affected by the change of pH in the range of 7.0 to 8.2. 4) The time for $half-hemloysis(t_{1/2})$ showed a temperature-dependency and Arrhenius plot exhibited a break point at $20^{\circ}C$. The apparent activation energy calculated from this plot was 18.1 kcal/mol between $2^{\circ}C-20^{\circ}C$ and 11.2 kcal/mol between $20^{\circ}C-37^{\circ}C$, respectively. 5) The apparent permeabilities of various inorganic anions based on $t_{1/2}$ were in the order of $Cl^->NO_{3}\;^->SCN^->SO_4\;^{2-}>SSO_3\;^{2-}>HPO_4\;^{2-}$. which was similar with the previous reports based on the experiment using radioisotopes. The results Obtained from this study are comparable with the previous data reported from the experiments using radioisotopes. This indicates that the hemolysis of erythrocytes in isoosmotic ammonium salt solution can be used as a simple and good method for the study of anion exchange in erythrocyte membrane.
Proceedings of the Technology Innovation Conference
/
1997.07a
/
pp.222-223
/
1997
By analyzing the successful innovation case of potato microtuber mass production technology, a representative case of technology-push type creative innovation in an imitation oriented research culture, this paper attempts to figure out conceptual model of creative innovation that is initiated by the public laboratories in catching-up country, Stages of creative innovation can be divided into the internal R&D stage and the external commercialization stage. Success of the internal R&D stage depended on autonomy to secure creative research idea and commitment of individual researchers. Psychological pressure evoked from sportlights of mass media and commitment of sponsor increased the intensity of research efforts of the researcher Recognition of research problem and its significance was intensified by site visits of agricultural fields, and the recognized higher impacts of expected research results and knowledge creation achieved were a fundamental source of self-motivation. In the stage of commercialization stage, various legal, socio-economic, and psychological barriers were confronted. In a catching-up country lacking of experiences of creative innovation, creative innovation process can be regarded as a barrier elimination and cultural revolution process. Among the barriers, psychological refusal of farmers to corn-sized potato seeds was critical, which finally enforced to further researches to enlarge the size of potato seeds. In addition, the researcher has concentrated his research efforts in one specialized research area by getting a series of similar research project funds rather than diversification. It was lucky for him to have a chance to carry out a series of similar researches in one research area during the last 10 years. In getting research funds from government and private companies continuously in one research area, both internal and external promoters played significant roles.
Background: It has been well known that bronchial asthma is a chronic inflammatory disorder. The "activation" of lymphocytes has a significant role in the pathogenesis of bronchial asthma. Among these lymphocytes, TH2-like rather than TH1-like lymphoytes are activated in the bronchial tissues from patients with atopic bronchial asthma. However, the difference of cytokines expression is not well documented between the atopic normal subjects and atopic asthmatics. Methods: Bronchial tissues were obtained from the tweleve atopic and non-atpoic asthmatics and tweleve atopic and non-atopic healthy subjects for in stiu hybridizatin of IL-2, IL-4, IL-5, and INF-$\gamma$. The probe of cytokines were tagged with digoxigenin by random priming method. Results: The infiltration of many inflammatory cells on submucosa and denuded epithelium were observed in the bronchial tissue from patients with bronchial asthma. The RNase-treated bronchial tissues did not have the brown signal on the tissue, but, RNasc-untreated bronchial tissues had the positive brown signal on the inflammatory cells under the basement membrane. The IL-2 positive signals were detected in 2 cases, IFN-$\gamma$ in 1 casc, IL-4 in 2 cases, IL-5 in 2 cases among 6 non-atopic healthy subjects. The atopic healthy subjects showed 1 case of positive signal of IL-2 and IFN-$\gamma$, but did not show any signals of IL-4 and IL-5. The positive signals of IL-2 were detected in 4 cases among 6 atopic and 6 non-atopic asthmatics, 2 cases and 1 case of IFN-$\gamma$ respectively, 4 cases and 3 cases of IL-4 respectively, 4 cases and 3 cases of IL-5 respectively. Conclusion: The lymphocytes were activated in the bronchus of asthmatics. Among lymphocytes, TH2-like lymphocytes may be involved in the pathogenesis of bronchial asthma. However, futher study with immunohistochemical stain may be necessary for defining the source of cytokines, because of TH2-like lymphocytes were also activated in some atopic healthy subjects.
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