Previously. we reported that the intraperitoneal administration of ginseng crude saponin increased: (I) nuclear RNA polymerase activity. (2) nuclear RNA synthesis. (3) cytoplasmic RNA synthesis. (4) cytoplasmic heavy polyrioosome content. (5) amino acid incorporation in vitro of microsome and polysome isolated rat liver. and (6) the incorporation rate of labeled amino acids into serum protein. In addition, a spectacular increase in the rough endoplasmic reticulum of hepatocyte administered crude saponin for four weeks orally was shown through electron microscopy. An increase in polysomal content in membrane-hound ribosome was shown through ultracentrifugation. Recently, successive intraperitoneal. administration .of $ginsenosid-Rb_2$ was given to streptozotocin (STZ) diaoetic rats of hypoproteinemia. The blood urea nitrogen and hepatic urea concentration were decreased significantly. The total protein and alhumin levels in the serum were increased in comparison to control values. In contrast. the $ginsenoside-Rb_2$ treated group of STZ diahetic rats showed a significant increase in liver RNA. total ribosome and membrane-bound ribosomal contents. The administration of $ginsenoside-Rb_2$ increased the incorporation rate of labeled - precursor into total serum protein. Additionally $ginsenoside-Rb_2$ improved the nitrogen balance of diabetic rats. On the bases of these experimental results, ginseng saponin has a metabolic stimulatory or anabolic action on RNA and protein synthesis.
The histochemical characteristics of the cutaneous granular glands and its fine structure of the Korean fire bellied toad, Bombino orientalis Boulenger the only species inhabitated in Korea, were studied with light and electron microscopes. By the granular size and histochemical properties, the granular glands were subdivided into two types. The type I glands have small secretory granules (3.6 $\mu$ m), whereas the type II glands contain relatively larger granules (7.5 $\mu$ m). These two types of glands were simple alveolar gland, and they were commonly composed of inner glandular epithelial cells and outermost myoepithelial cells. By the histochemical reactions, these two secretory granules were not responsed to Alcian blue, but weakly positive reaction was observed in type I granular glands by the PAS reaction. And by the scanning electron microscopic observation, extemal morphology of the opening site of the gland duct was resembled to the stomata of the plant leaves. Especially the epithelial cells of the type I glands had fine reticular surface whereas, type II glands had large vesicular surface. These types of the glands conimonly had larger cavities than those of mucous glands, and forming the multinucleated unicellulr structure (syncytium) at the glandular epithelial layers. Secretory granules of these glands were originated from the rough endoplasmic reticulum of the glandular epithelial cells, and by the maturation level of the granule, morphological diversities appeared.
Paraquat, a widely used herbicide, has been suggested as a potential risk factor for Parkinson's disease. Heme oxygenase-1(HO-1), a marker for oxidative stress and endoplasmic reticulum(ER) stress, is known to catalyze heme to biliverdin, carbon monoxide and free iron in response to various stimuli. Here we show that paraquat activates HO-1 expression in a time-and dose-dependent manner in substantia nigra(SN) dopaminergic neuronal cells. Activation of Ho-1 by paraquat was regulated primarily at the level of gene transcription. Deletion analysis of the promoter and the 5' distal enhancers, E1 and E2, of the HO-1 gene revealed that the E2 enhancer is a potent inducer of the paraquat-dependent Ho-1 gene expression in dopamninergic neuronal cells. Mutational analysis of the E2 enhacer further demonstrated that the transcription factor activator protein-1(AP-1) plays an important role in mediating paraquat-induced HO-1 gene transcription. Moreover, using specific inhibitors of the mitogen-activated protein kinases(MAPKs), we investigated the role of paraquat and MAPKs for HO-1 gene regulation in dopaminergic cells. The c-Jun N-terminal kinase(JNK) inhibitor SP600125 significantly suppressed the expression of HO-1 by paraquat. All these results demonstrate that induction of HO-1 by paraquat requies the activation of the AP-1 and JNK pathway.
In the present study, the vacuolar apparatus were investigated in the hepatocytes of rats treated with DA by transmission electron microscopy of conventional and cytochemical thin sections. In the rats after 20 min of dehydrocholic acid treatment, the cis Golgj cisterns were sacculated in line. The saccule occasionally occured by elongation and attenuated neck. The lysosomes also showed protrudent saccule. The vesicles were observed near the cis Golgi cisterns, lysosome and bile canaliculi. Some of the vesicles appeared to be fused to bile canaliculi. The cis Golgi cisterns usually faced toward the bile canaliculi both in normal and experimental groups. The cis Golgi cisterns, protrudent saccule and vesicles were almost devoid of visible contents. The osmium deposits were heavy on the protrudent saccule as well as on the cis Golgi cisterns or on the vesicles isolated near by, but they were light or not observed on the vesicles in the immediate vicinity of bile canaliculi. The acid phosphatase activities appeared on the lysosome and vesicles located near by, but did not appear on the vesicles as approaching closer to the bile canaliculi. The evidence suggests that the vesicles are derived from the cis Gogi cistern and lysosomes and fuse to bile canaliculi for exocytosis, and that the activity in the vesicles is diminished as approaching closer to the bile canaliculi.
The aim of the present study was to examine in detail, both at light and electron microscopical levels, the morphological variations in ameloblast of the fetal rat incisor enamel organ. Rats were started on distilled water at the beginning of pregnancy. The pups were sacrificed 11 days after delivery and animals were perfused intravascularly with glutaraldehyde and the incisors were removed. To examine on the ultrastructure of the ameloblast, the study employed primary light microscopy but electron microscopy was used to clarify some of the light microscopic finding. Longitudinal sections through the incisors of the rat show a continuous layer of ameloblasts on the labial surface of the tooth. This layer contains the entire sequence of developmental stages in enamel production. The ameloblast layer was divided into three main zones: 1) Presecretory zone, region of ameloblasts facing pulp. 2) Secretory zone, region of inner and outer enamel secretion. 3) Maturation zone, region of reduced ameloblasts. In particularly, the present study has shown that two distinctively different types of ameloblasts appear in the enamel organ during enamel maturation in the rat incisor. These two types have been designated ruffle-ended ameloblasts (rAB) and smooth-ended ameloblasts (sAB). The fluoride produces marked alteration in the fine structure of ameloblast from teeth of young rats, such as large confluent distensions of the endoplasmic reticulum and swelling of isolated mitochondria. This experimental data suggested that exposure prolonged of animal to high level of fluoride appears to induce a few dramatic changes in the normal appositional growth and initial mineralization of enamel created during amelogenesis.
The development of the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm of crown-rump length (10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cell, primitive neuroblast, primitive supporting cell, and unmyelinated fibers. At 70mm fetus, the neuroblasts and their processes were ensheated by the bodies or processes of satellite cells. The cytoplasm of the neuroblast contained rough endoplasmic reticulum, mitochondria, Golgi complex, Nissl bodies and dense-cored vesicles. As the neuroblasts grew and differentiated dense-cored vesicles moved away from perikaryal cytoplasm into developing processes. Synaptic contacts between the cholinergic axon and dendrites of postganglionic neuron and a few axosomatic synapses were first observed at 70 mm fetus. At 90 mm fetus the superior cervical ganglion consisted of neuroblasts, satellite cells, granule-containing cells, and unmyelinated nerve fibers. The ganglion cells increased somewhat in numbers and size by 150 mm fetus. Further differentiation resulted in the formation of young ganglion cells, whose cytoplasm was densely filled with cell organelles. During next prenatal stage up to 260 mm fetus, the cytoplasm of the ganglion cells contained except for large pigment granules, all intracytoplasmic structures which were also found in mature superior cervical ganglion. A great number of synaptic contact zones between the cholinergic preganglionic axon and the dendrites of the postganglionic neuron were observed and a few axosomatic synapses were also observed. Two morphological types of the granule-containing cells in the superior cervical ganglion were first identified at 90 mm fetus. Type I granule-containing cell occurred in solitary, whereas type II tended to appeared in clusters near the blood capillaries. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma of type I granule-containing cells and preganglionic axon termials were observed. In addition, synaptic junctions between the processes of the granule- containing cells and dendrites of postganglionic neuron were also observed from 150 mm fetus onward. In conclusion, superior cervical ganglion cells and granule-containing cells arise from a common undifferentiated cell precursor of neural crest . The granule-containg cells exhibit a local modulatory feedback system in the superior cervical ganglion and nay serve as interneurons between the preganglionic and postganglionic cells.
Fluoxetine, a widely used anti-depressant compound, has several additional effects, including blockade of voltage-gated ion channels. We examined whether fluoxetine affects ATP-induced calcium signaling in PC12 cells by using fura-2-based digital calcium imaging and assay for $[^3H]-inositol$ phosphates (IPs). Treatment with ATP $(100\;{\mu}M)$ for 2 min induced $[Ca^{2+}]_i$ increases. The ATP-induced $[Ca^{2+}]_i$ increases were significantly decreased by removal of extracellular $Ca^{2+}$ and treatment with the inhibitor of endoplasmic reticulum $Ca^{2+}$ ATPase thapsigargin $(1\;{\mu}M)$. Treatment with fluoxetine for 5 min blocked the ATP-induced $[Ca^{2+}]_i$ increase concentration-dependently. Treatment with fluoxetine $(30\;{\mu}M)$ for 5 min blocked the ATP-induced $[Ca^{2+}]_i$ increase following removal of extracellular $Ca^{2+}$ and depletion of intracellular $Ca^{2+}$ stores. While treatment with the L-type $Ca^{2+}$ channel antagonist nimodipine for 10 min inhibited the ATP-induced $[Ca^{2+}]_i$ increases significantly, treatment with fluoxetine alone blocked the ATP-induced responses. Treatment with fluoxetine also inhibited the 50 mM $K^+-induced$$[Ca^{2+}]_i$ increases completely. However, treatment with fluoxetine did not inhibit the ATP-induced $[^3H]-IPs$ formation. Collectively, we conclude that fluoxetine inhibits ATP-indueed $[Ca^{2+}]_i$ increases in PC12 cells by inhibiting both an influx of extracellular $Ca^{2+}$ and a release of $Ca^{2+}$ from intracellular stores without affecting IPs formation.
This study is to characterize the postulated anti-inflammatory effect of the hot water extracts from the Phellinus baumii. RAW264.7, macrophage cell line, was activated by lipopolysaccharide (LPS) and, further, treated with Phellinus baumii's aqueous extract. When the cultured macrophage cells were treated with LPS, they show typical signs of endoplasmic reticulum stress (ERS) and an increment in secretion of inflammatory cytokine compared to the non-treated control: The expression of glucose-regulated protein78 (Grp78), Grp94, and C/EBP homologous protein/GADD 153 (CHOP) increased along with augmented secretion of interlukin-6. Cellular nitric oxide content also significantly went up in comparison to the non-LPS treatment. When the LPS-treated RAW264.7 was treated with the aqueous Phellinus baumii extracts, however, the expression of ERS markers markedly reduced and the release of nitric oxide declined. Also, the expression of induced nitric oxide synthase (iNOS) notably diminished similarly as the NO content. In conclusion, this study strongly indicated that aqueous Phellinus baumii extract can be utilized directly as anti-inflammation agent and serves as a source of functional ingredient to lessen the inflammation.
The present study was carried out to examine the effect of a calcinogen, aflatoxin B1 on the ultrastructural changes of ciliary epithelial cells in mice infected with Clonerchis sinensis. A total of 93 male albino mice(BALB/c strain) was divided into 3 groups; group I, treated with 1.0 ppm aflatoxin Bl for 12 weeks; group II, given 50 C. sinensis n;etacercariae, and group III, given 50 metacercariae and treated with 1.0 ppm aflatoxin Bl for 12 weeks. Three mice served for untreated-uninfected controls. From 4 weeks after the treatsment and/or in(ection, three mice from each group were sacrificed at 4 week intervals up to the 40th week, and their hepatobiliary tissues were prepared for transmission electron microscopy. The most prominent ultrastructural changes in group I were remarkable enlargement of nuclear size, separation of nucleolus, dispersed chromatin granules in nuclei and increased dense granules along the inner membrane of nuclei. In the cytoplasm there was slight proliferation of mitochondria and endoplasmic reticulum (ER) at earlier stage. At the 12th week separation of fibrillar and granular components of the nucleolus was a characteristic finding. As the time elapsed, epithelial cells showed fiattened-cuboidal form and a tendency of atrophy. Most of the nuclei were elongated and polygonal in shape. In group II the appearance of elaborate interwoven folds of lateral cytoplasm forming a labyrinth of interconnected intercellular space and variety in nuclear shape were the prominent fadings at earlier stage. The cytoplasm showed slight proliferation and dilatation of mitochondria and ER, and a small number of mucin droplets. In the basement membrane scanty fibrous cells were seen. With time, variety in nuclear shape, marked proliferation and dilatation of rough ER and some collagen fibrils were demonstrated. Other features of intracellular organelles and mucin droplets persisted. In group III cuboidal epithelial cells showed their remarkably enlarged and irregular nuclei, increased chromatin granules in the nuclei, separated nucleoli, proliferated and dilated rough ER. With time, sequestered mitochondria showed blob-like evaginations which lacked cristae and dense matrix, and were limited by a single membrane. Since the 20th week, microvilli were relatively scanty and poorly developed. Organelles and inclusions in the cytoplasm of metaplastic cells were poor. Nuclei were variable in shape. The nlost prominent changes at later stage were separation of nuclei from the cytoplasm, and appearance of numerous and irregularly angled electron dense granules in the nuclei.
Saturated fatty acids contribute to β-cell dysfunction in the onset of type 2 diabetes mellitus. Cellular responses to lipotoxicity include oxidative stress, endoplasmic reticulum (ER) stress, and blockage of autophagy. Palmitate induces ER Ca2+ depletion followed by notable store-operated Ca2+ entry. Subsequent elevation of cytosolic Ca2+ can activate undesirable signaling pathways culminating in cell death. Mitochondrial Ca2+ uniporter (MCU) is the major route for Ca2+ uptake into the matrix and couples metabolism with insulin secretion. However, it has been unclear whether mitochondrial Ca2+ uptake plays a protective role or contributes to lipotoxicity. Here, we observed palmitate upregulated MCU protein expression in a mouse clonal β-cell, MIN6, under normal glucose, but not high glucose medium. Palmitate elevated baseline cytosolic Ca2+ concentration ([Ca2+]i) and reduced depolarization-triggered Ca2+ influx likely due to the inactivation of voltage-gated Ca2+ channels (VGCCs). Targeted reduction of MCU expression using RNA interference abolished mitochondrial superoxide production but exacerbated palmitate-induced [Ca2+]i overload. Consequently, MCU knockdown aggravated blockage of autophagic degradation. In contrast, co-treatment with verapamil, a VGCC inhibitor, prevented palmitate-induced basal [Ca2+]i elevation and defective [Ca2+]i transients. Extracellular Ca2+ chelation as well as VGCC inhibitors effectively rescued autophagy defects and cytotoxicity. These observations suggest enhanced mitochondrial Ca2+ uptake via MCU upregulation is a mechanism by which pancreatic β-cells are able to alleviate cytosolic Ca2+ overload and its detrimental consequences.
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