Immune responses of periodontal tissue may be regulated by products of sensory afferent nerve endings such as neuropeptides. Substance P(SP), a tachykinin neuropeptide, has been previously reported to stimulate the activities of T lymphocyte. Therefore, I examined the role of SP in IL-2 production and cell proliferation by using a homogeneous line of T lymphocytes(Jurkat and HuT78). Cell proliferation rate was determined by [$^3H$]-thymidine incorporation test, and IL-2 was quantitated by the growth rate of CD4+ IL-2-dependent T lymphocyte line CTLL-2. SP stimulated cell proliferation of T lymphocytes at the concentration of $10^{-12}$ and $10^{-8}$M in a biphasic bell-shape dose-dependent manner. However, SP alone did not induce IL-2 release at the concentration range of $10^{-6}$ to $10^{-14}$M. The upregulation of IL-2 release was observed when $10^{-12}$M SP was applied together with mitogens such as Con A or PHA+PMA on T cell lines, especially on Jurkat. Con A or PHA+PMA demonstrated to increase the rate of cell proliferation of Jurkat, which had shown to produce much amount of IL-2 indicating that mitogen-induced cell proliferation might be partially influenced by released IL-2. It was concluded that regulatory effects of SP on the immune/inflammatory response could be mediated through the costimulatory upregulation of IL-2 production and increase of cell proliferation of T lymphocyte.
Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca2+ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca2+-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca2+ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.
A study was undertaken to observe the distribution and ultrastructure of the sensory papillae of the liver and lung flukes which was obtained from experimentally infected rats and dogs. For this study, the rats were artificially infected with metacercariae isolated from Pseudorasbora parve, and the dogs were artificially infected with metacercariae isolated from Cambaroides similis. The liver flukes (Clonorchis sinensis) were collected from the bile ducts of the rats which were autopsied 5 weeks later since infection. The lung flukes (Paragonimus westermani) were collected from the lung of the dogs which were autopsied 3 months later since infection. With the collected worms, ultrastructure of sensory papillae was studied by means of a scanning and transmission electron microscope. The liver flukes were compared with the lung flukes. The results of the observation are as follows. 1. The sensory papillae of the liver flukes was classified into 3 types and most of sensory papillae were distributed on the oral sucker. 2. The sensory papillae of the lung flukes were distinguished 8 types. The sensory papilla type VIII was specifically distributed on the oral sucker and type I, II were distributed on the tegumental surface of the worm. The sensory papillae of the lung flukes were mostly observed between oral sucker and ventral sucker of the worm excusively 3. The sensory papillae of the liver flukes were distributed around the sucker while those of the lung flukes were developed between spine. 4. The sensory papillae were formed by many tegumental ridge in the liver flukes while was made of many small sensory papillae in the lung flukes. 5. The sensory cilia were observed between tegumental ridges in the sensory papilla of the liver flukes and also in lung flukes. And they are alike in that respect. 6. The sensory papillae were not developed in the posterior part of the liver and lung flukes but mostly nerve endings were formed in the tegumental syncytium, and it was connected with neuropile.
The neuropeptide substance P(SP) has been recognized to modulate immune systems, with close proximity between peptidergic sensory nerve endings and immune cells. These include the macrophage and neutrophil activation, IL-2 production in T cell, augmentation of Ig synthesis, mast cell degranulation, $PGE_2$ and collagenase secretion in synoviocytes. In this study I examined SP-induced various biological activities such as antimicrobial action, cytokine production, and mast cell degranulation in the presence or absence of other inflammatory cell activators. Antimicrobial studies showed that undifferentiated HL-60 cells were not affected by SP. However, SP significantly enhanced antimicrobial action of TPA-treated or dbcAMP-treated HL-60 cells which had been differentiated into PMN or macrophage/monocyte. I could not find synergistic relationship between SP and LPS in parallel experiments of the above. SP did not induce IL-l production from murine macrophage cell line RAW264.7 whether costimulated with LPS or not. Mast cell degranulation was occured only when stimulated with high dose ($10^{-5}M$) of SP and the degree of this activation was slightly reduced by simultaneous application of $MIP-1{\alpha}$. In addition, CGRP which is known to be a common coexisting neuropeptide with SP within specific fibers did not augment the function of SP on mast cell degranulation. These results suggest that immunoregulatory activities of SP could be mediated through direct upregulation of various functions of immune cells and also upregulation of responsiveness of immune cells to other immune activators.
Optic lobes of Todarodes pacificus and Octopus minor are largely divided into cortex and medulla, the cortex being composed of three layers (an outer granule cell layer, a plexiform layer, and an inner granule cell layer). The cortex of Todarodes pacificus is about $420{\sim}450{\mu}m$ thick, being $170{\sim}200{\mu}m$ thicker than that of Octopus minor of which thickness is about $250{\sim}290{\mu}m$. In the outer granule cell layer of Todarodes pacificus, three types of nerve cells (type-A, type-B and type-C) and neuroglial cells that surround or contact with the neurons are observed, while in the outer granule cell layer of Octopus minor, two types of nerve cells (type-A and type-B) and a single type of neuroglial cells are observed. In a plexiform layer, a presynaptic bag and nerve endings are connected to each other, consequently forming various types of synaptosomes. The synaptosomes of Todarodes pacificus contain electron dense vesicles, electron dense-core vesicles and electron lucent vesicles, either individually or in a mixture. On the other hand, three types of synaptosomes a mixture of electron dense-core vesicles and electron lucent vesicles, electron lucent vesicles only, and electron dense-core vesicles only are observed in Octopus minor. The structures of the inner granule cell layer are almost similar in the two species. It is composed of two types of nerve cells (type-A, type-B) and a single type of neuroglial cells. In the medulla of Todarodes pacificus, the cells of $7{\times}5{\mu}m$ are arranged to a line and form the palisade cell layer, but these are not observed in Octopus minor.
In order to evaluate the mechanism of transmission as well as processing of sensory information originating from low-threshold mechanoreceptor in oral and maxillofacial region at primary synaptic region of trigeminal nervous system, vibrissa afferent fibers of adult cat were labeled with intra-axonal HRP injection. Serial sections containing labeled boutons were obtained from the piece of trigeminal interpolar nucleus. Under electron microscope, total 30 labeled boutons were observed, and ultrastructural characteristics, frequency of occurence, synaptic organizations of vibrissa afferent terminals were analysed. The results were as follows: 1. Labeled boutons contained clear, spherical synaptic vesicles with diameter of 45$\sim$55nm. They formed asymmetrical synapse with dendrites showing definite postsynaptic density, larger synaptic cleft, multiple synaptic structures at various regions. With unlabeled axon terminals(p-ending) containing polymorphic synaptic vesicles, they formed symmetrical synapse showing indefinite postsynaptic density and narrower synaptic area. 2. Each labeled bouton formed 1 to 15 synapses, the average of 4.77$\pm$3.37 contacts per labeled bouton, with adjacent neuronal profiles. Relatively complex synaptic organization, which formed synapses with more than 5 neuronal profiles, was observed in a large number(46.7%, n=14) of labeled boutons. 3. Axo-somatic synapse was not observed. The number of axo-dendritic synapse was 1.83$\pm$1.37 per labeled bouton. Majority(85.0%) of axo-dendritic synapses were formed with dendritic shafts, nonprimary dendrites(n=47, 1.57$\pm$1.38/1 bouton), however, synapses formed with primary dendrites(n=6, 0.20$\pm$0.41/1 bouton) or dendritic spines(n=2, 0.07$\pm$0.25/1 bouton) were rare. 4. 76.7%(n=23) of labeled boutons formed axo-axonic synapse (2.93$\pm$2.36/1 bouton) with p-endings containing pleomorphic vesicles. Synaptic triad, in which p-endings formed synapses with labeled boutons and dendrites adjacent to the labeled boutons simultaneoulsy, were also observed in 60.0%(n=18) of labeled boutons. From the above results, vibrissa afferent terminals of adult cat showed distinctive synaptic organization in the trigeminal interpolar nucleus, thus, suggests their correlation with the function of the trigeminal interpolaris nucleus, which participates in processing of complex sensory information such as two-point discrimination and motivational-affective action. Further studies on physiologic functions such as quantitative analysis on ultrastructures of afferent terminals and nerve transmitters participating in presynaptic inhibition are required.
Xerostomia and xerophthalmia are delicate or serous side effects, occuring when the radiotherapy is administered to the head and neck cancer patient. It is known that the cause of the above side effect is radiosensitivity of serous cells. In this study, the ultrastructural features of the parotid glands of the X-irradiated rats were observed. Sprague-Dawley rats weighing 200-250g each were anesthetized with sodium thiopental, and placed on the Mitsubishi linear accelerator. Only the head and neck areas of animals were exposured at the distance of 80cm, within the area of $30X30cm$, in the depth of 1cm, with the speed of 200R/min. Total doses applied were 3,000R or 6,000R depending on the experimental groups. Animals were sacrificed on the 6th hour, 2nd day and 6th day after the irradiation. Parotid glands were fixed in the 2.5% glutaraldehyde-1.5% paraformaldehyde solution, and followed by refixation in the 1% osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture, and ultrathin sections were cut. Sections were contrasted with the solution of uranyl acetate and lead citrate, and observed with JEM 100 CX-II electron microscope. The results were as follows: 1. Normal parotid acinar cells are two types; the light and the dark acinar cells. The light acinar cell contains dense secretory granules, whereas dark acinar cells contains granules of medium density with some darker spots within them, or other cells contain granules of medium density with darker rims. 2. Six hours after the irradiation, many acinar cells were degenerated showing variable stages of cytolytic bodies, light bodies, or dense degenerations. Within the acinar cell, Golgi apparatus and granular endoplasmic reticula were most severely altered elements. Granules showed more contrasting densities and irregularities. 3. Two days after the irradiation, some cytolytic bodies, and focal lucent degeneration of cytoplasm, and fine granular alteration of cytoplasmic matrix were pronounced. But other elements including secretory granules are rather looked unlatered. 4. Six days after the irradiation, most severe alterations were seen. Many intracellular canaliculi (or secretion figures), quanta of cytoplasm containing secretion antecedants, severely irregular luminal border, and again contrasting density of secretory granules showing tigroid spots or dense rims were noted. And myoepithelial degenerations were observed not uncommonly. 5. Irregular densities of secretory granules were interpreted as abnormal components of protein or carbohydrate portion are synthesized or abnormally metabolized under severe X-irradiation. 6. Myoepithelial degeneration and related alteration of nerve endings, etc., were suggested as the other causes of xerostomia following X-irradiation. 7. It is requested that radiation doses should be arranged, considering in mind not only the sensitivity of acinar cells but also the myoepithelial and neural functions.
Purpose: Hands are the chief organs for physically manipulating the environment, using anywhere from the roughest motor skills to the finest, and since the fingertips contain some of the densest areas of nerve endings on the human body, they are continuously used organ with complex functions, and therefore, often gets injured. To prevent any functional loss, a detailed anatomical knowledge is required to have a perfect surgical treatment. Also it is necessary to have a thorough understanding of arrangements of the human extensor tendons and intertendinous connections when tenoplasty or tendon transfer is required. We performed a study of the arrangements of the human extensor tendons and the configuration of the intertendinous connections over the dorsum of the wrist and hand. Methods: A total of 58 hands from Korean cadavers were dissected. The arrangements of extensor indicis proprius, extensor digitorum communis, and extensor digiti minimi tendons and intertendinous connections were studied. Results: The most common distribution patterns of the extensor tendons of the fingers were as follows: a single extensor indicis proprius (EIP) tendon which inserted ulnar to the extensor digitorum-index (EDC-index); a single EDC-index; a single EDC-middle; a double EDC-ring; an absent EDC-little; a double extensor digiti minimi (EDM), a single EDC-index (98.3%), a single EDC-middle (62%), a double EDC-ring (50%), and an absent (65.5%) or a single (32.8%) EDC-little. A double (70.6%) EDM tendons were seen. Intertendinous connections were classified into 3 types: type 1 with thin filamentous type, type 2 with a thick filamentous type, and type 3 with a tendinous type subdivided to r shaped 3r type and y shaped 3y type. The most common patterns were type 1 in the 2nd intermetacarpal space, type 2 in the 3rd intermetacarpal space, and type 3r in the 4th intermetacarpal space. And in the present study, we observed one case of the extensor digitorum brevis manus (EDBM) on the boht side. Conclusion: A knowledge of both the usual and possible variations of the extensor tendon and the intertendinous connection is useful in the identification and repair of these structures.
Journal of the Korea Academia-Industrial cooperation Society
/
v.18
no.1
/
pp.295-301
/
2017
Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.
Song, Dae Jin;Song, Woo-Jung;Kwon, Jae-Woo;Kim, Gun-Woo;Kim, Mi-Ae;Kim, Mi-Yeong;Kim, Min-Hye;Kim, Sang-Ha;Kim, Sang-Heon;Kim, Sang Hyuck;Kim, Sun-Tae;Kim, Sae-Hoon;Kim, Ja Kyoung;Kim, Joo-Hee;Kim, Hyun Jung;Kim, Hyo-Bin;Park, Kyung-Hee;Yoon, Jae Kyun;Lee, Byung-Jae;Lee, Seung-Eun;Lee, Young Mok;Lee, Yong Ju;Lim, Kyung-Hwan;Jeon, You Hoon;Jo, Eun-Jung;Jee, Young-Koo;Jin, Hyun Jung;Choi, Sun Hee;Hur, Gyu Young;Cho, Sang-Heon;Kim, Sang-Hoon;Lim, Dae Hyun
Allergy, Asthma & Immunology Research
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v.10
no.6
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pp.591-613
/
2018
Chronic cough is common in the community and causes significant morbidity. Several factors may underlie this problem, but comorbid conditions located at sensory nerve endings that regulate the cough reflex, including rhinitis, rhinosinusitis, asthma, eosinophilic bronchitis, and gastroesophageal reflux disease, are considered important. However, chronic cough is frequently non-specific and accompanied by not easily identifiable causes during the initial evaluation. Therefore, there are unmet needs for developing empirical treatment and practical diagnostic approaches that can be applied in primary clinics. Meanwhile, in referral clinics, a considerable proportion of adult patients with chronic cough are unexplained or refractory to conventional treatment. The present clinical practice guidelines aim to address major clinical questions regarding empirical treatment, practical diagnostic tools for non-specific chronic cough, and available therapeutic options for chronic wet cough in children and unexplained chronic cough in adults in Korea.
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