Therapeutic potential of sulforaphane: modulation of NRF2 mediated PI3/AKT/mTOR pathway in oral fibrosis / Potencial terapêutico do sulforafano: modulação do NRF-2 mediado o pela via PI3/AKT/mTOR na fibrose oral

Oral Submucous Fibrosis is a potentially malignant disorder caused by habitual areca nut chewing, which contributes to the dispersion of active alkaloids into subepithelial tissues, stimulating excessive extracellular matrix deposition. Various treatment modalities are available; however, their efficacy in inhibiting fibrosis progression remains limited. Sulforaphane (SFN), an isothiocyanate found abundantly in cruciferous plants, is known to have effective antifibrotic properties. Objective: The present study investigated the antifibrotic effect of SFN via phosphatidylinositol 3 kinase (PI3K), Serine/Threonine Kinase 1 (AKT-1), mammalian target of rapamycin (mTOR) pathway in arecoline (AER) induced fibrosis in human gingival fibroblasts [HGFs]. Material and Methods: MTT assay determined the half-maximal inhibitory concentration of AER and SFN at 24h in the HGF cell line. Expression levels of transforming growth factor ß1 (TGFß1), collagen type 1 alpha 2 (COL1A2), hydroxyproline (HYP), PI3, AKT, mTOR, and nuclear factor erythroid 2­related factor 2 (NRF2) were assessed post-AER and SFN treatment using qPCR and western blot analysis. Results: The findings of the study revealed that AER elicited a stimulatory effect, upregulating TGFß1, COL1A2, HYP, PI3K, AKT, and mTOR and downregulating NRF2 expression. Conversely, SFN treatment significantly upregulated NRF2, inhibiting TGFß1 mediated PI3/AKT/mTOR pathway. Conclusion: These observations suggest that SFN can be used as a promising synergistic antifibrotic agent to combat fibrogenesis via the non-Smad pathway (AU)

Fibrose submucosa oral é uma desordem potencialmente maligna causada pelo habito de mascar a noz da areca, o que contribui para a dispersão de alcalóides ativos nos tecidos subepiteliais, estimulando a deposição excessiva de matriz extracelular. Há várias modalidades terapêuticas, no entanto, com eficácia limitada no controle da progressão da fibrose. O sulforafano (SFN), isotiocianato encontrado abundantemente em plantas crucíferas, é conhecido por suas propriedades antifibróticas. Objetivo: Investigar os efeitos antifibróticos do SFN na via fosfatidilinositol3-quinase (PI3K), via quinase serina/treonina 1 (AKT-1), via do alvo da rapamicina em mamíferos (mTOR), na fibrose induzida por arecolina (AER) em fibroblastos gengivais de humanos (HGFs). Material e Métodos: A meia concentração inibitória mínima de AER e SFN em 24 horas nas células HGFs foi determinada por MTT. Os níveis de expressão de ß1 (TGFß1), colágeno tipo 1 alfa 2 (COL1A2), hidroxiprolina (HYP), PI3K, AKT, mTOR, fator nuclear eritroide 2 relacionado ao fator 2 (NRF2) foram analisados após tratamento com ERA e SFN através de qPCR e western blot. Resultados: O ERA apresentou efeito estimulatório aumentando a expressão de TGFß1, COL1A2, HYP, PI3K, AKT e mTOR e diminuindo a expressão de NRF2. Por outro lado, tratamento com SFN aumentou significativamente a expressão de NRF2, inibindo a liberação de TGFß1 mediada pela via PI3/AKT/mTOR. Conclusão: Esses achados sugerem que o SFN pode ser um agente antifibrótico promissor no combate à fibrogênese decorrente da via não-Smad (AU)

Arecoline induces activation of human oral fibroblasts by promoting macrophage secretion of exosomes containing miR-155-5p / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which arecoline regulates the level of miR-155-5p in macrophage-secreted exosomes to induce the transformation of human oral mucosal fibroblasts (HOMFs) into fibroblast phenotype.@*METHODS@#Exosomes were harvested from human monocytic cell line THP-1 with or without arecoline treatment. The effects of arecoline-treated THP-1 cell culture supernatant (CS), THP-1-derived exosomes (EXO), exosome-depleted THP-1 cell supernatant (NES), miR-155-5p overexpression, and miR-155-5p inhibitor on migration ability of arecoline-treated HOMF cells were examined using Transwell migration assay. The polarization of THP-1 cells was detected using flow cytometry. DCFH-DA was used to detect the level of oxidative stress in the cells with different treatments. The mRNA and protein expressions of α- SMA, type I collagen and SOCS1 in the cells were detected with qRT-PCR and Western blotting.@*RESULTS@#Flow cytometry showed that arecoline-treated THP-1 cells exhibited obvious polarization from M0 to M1. Both the supernatant and exosomes from arecoline-treated THP-1 cells significantly enhanced the migration ability of HOMF cells, increased intracellular oxidative stress, up-regulated the expressions of miR-155- 5p and the mRNA and protein levels of α-SMA and type I collagen, and lowered the mRNA and protein expressions of SOCS1. In HOMF cells treated with exosomes from arecoline- treated THP-1 cells, overexpression of miR-155-5p significantly enhanced cell migration ability and increased cellular expressions of α-SMA and type I collagen, and miR-155-5p inhibitor caused the opposite changes.@*CONCLUSION@#Arecoline can up-regulate miR-155-5p expression in THP-1 cells and inhibit the expression of SOCS1 protein in HOMF cells via the exosome pathway, thus promoting the fibrotic phenotype transformation of HOMF cells.

Water migration and kinetics of Arecae Semen during moistening process / 中国中药杂志

In this study, low-field nuclear magnetic resonance(LF-NMR) and magnetic resonance imaging(MRI) were employed to analyze the water distribution, status, and migration in the moistening process of Arecae Semen. Peleg model was adopted to study the water absorption kinetics of Arecae Semen moistened at different water temperatures(10, 30, and 50 ℃). The Arecae Semen samples soaked at different water temperatures all contained four water states: binding water T_(21), non-flowing water T_(22), free water T_(23), and unbound water T_(24). Non-flowing water had the largest increase in peak area during the moistening process, followed by free water. The peak areas of non-flowing water, free water, and total water were correlated with the water content(P<0.01). Therefore, LF-NMR can quickly and non-destructively predict the water content of Arecae Semen during moistening. The peak area of non-flowing water and the content of free water were correlated with the content of arecoline in the soaking solution(P<0.01), which indicated that the faster flow of non-flowing water and more free water corresponded to more arecoline dissolved. The MRI images showed that the water migration pathway varied at different soaking temperatures, and the moistening degree obtained by this means was consistent with that obtained based on traditional experience. The rate constant K_1 fitted by Peleg model decreased with the increase in water temperature, while the capacity constant K_2 showed an opposite trend. The Arrhenius equation fitting of K_1 with temperature showed that the activation energy of Arecae Semen in the moistening process was 32.98 kJ·mol~(-1). LF-NMR/MRI can be used to analyze the water status and content and determine the end moisturing point of Arecae Semen. Peleg model can accurately describe the water absorption properties of Arecae Semen in the moistening process. The findings of this study can guide the moistening optimization and mechanism research of other seed Chinese medicinal materials.

Rat model with oral submucous fibrosis induced by arecoline and mechanical stimulation / 华西口腔医学杂志

OBJECTIVE@#The aim of this study was to induce oral submucous fibrosis (OSF) in Sprague-Dawley(SD) rat models by arecoline and mechanical stimulation.@*METHODS@#Two factors factorial design was used to divide 48 rats into 8 groups (n=6). Different concentrations of arecoline (0, 0.5, 2, and 8 mg·mL⁻¹) and mechanical stimulation (with or without brush) were treated. After 16 weeks of treatment, the mouth opening was measured, the pathological changes of the buccal mucosa were observed, and the expressions of type Ⅲ collagen, transforming growth factor β1 (TGF-β1), and interferon-γ (IFN-γ) were detected.@*RESULTS@#In rats with moderate and high concentrations of arecoline, typical OSF pathological features were observed in the buccal mucosa, the mouth openings were significantly reduced, and the expression levels of type Ⅲ colla-gen and TGF-β1 were significantly increased (P0.05).@*CONCLUSIONS@#Moderate and high concentrations of arecoline can induce OSF in SD rats, but mechanical stimulation cannot induce OSF.

Induction of rat hepatic CYP2E1 expression by arecoline in vivo / 药学学报

The regulation mechanism of arecoline on rat hepatic CYP2E1 was studied in vivo. After oral administration of arecoline hydrobromide (AH; 4, 20 and 100 mg x kg(-1) x d(-1)) to rats for one week, the hepatic CYP2E1 mRNA level remained unchanged, but the hepatic CYP2E1 protein content was dose-dependently increased. Additionally, although the hepatic CYP2E1 activity was induced by AH treatment, the induction was attenuated with the increase in dosage. The results indicate that the effect of arecoline on rat hepaticdoes not involve transcriptional activation of the gene, but largely involves the stabilization of CYP2E1 protein against degradation or increased efficiency of CYP2E1 mRNA translation, and additionally involve the post- ranslational modification of CYP2E1 protein. Furthermore, the CYP2E1 response is fairly equal among the different species, the induction of rat hepatic CYP2E1 by arecoline suggests that there is a risk of metabolic interaction among the substrate drugs of CYP2E1 in betel-quid use human.

Changes of miRNA after oral submucous fibrosis co-cultured with Salvia and low-dose prednisolone / 中南大学学报(医学版)

OBJECTIVE@#To explore and analyze the the expression change of miRNA associated with oral submucous fibrosis (OSF) treated by the Salvia combined with law-dose prednisolone.@*METHODS@#Ten pairs of tissues from patients with typical early or advanced stage clinical pathological features of OSF and their paired normal tissues (internal control), were selected respectively. The miRNA expression profiles between the OSF and its paired controls were compared by the Affymetrix analysis. The primary normal oral mucous cells were cultured in arecoline (50 μg/mL) for 3, 6, 12 d (0 d ser ved as cont rol), and the primary OSF-fibroblast cells were cultured with Salvia (90 mg/mL) combined with low-dose prednisolone for 12, 24, 36 h (0 h served as control). The differential expression of miRNA was detected.@*RESULTS@#Arecoline induced the expression changes of miRNAs in normal mucosal cells. Salvia combined with low doses of prednisolone reversed the related miRNA expression.@*CONCLUSION@#MiRNAs play an essential role in the occurrence and development of OSF. Salvia combined with low-dose prednisolone can reverse the expression of related miRNAs in OSF cells.

Stimulation of endothelial non-neuronal muscarinic receptor attenuates the progression of atherosclerosis via inhibiting endothelial cells activation / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of non-neuronal muscarinic receptors (NNMR) stimulation on atherosclerosis and endothelial cells activation.</p><p><b>METHODS</b>Atherosclerosis model was established in ApoE-/- mice by a high fat diet for 7 weeks. During the experimental periods, animals were received a low (7 mg/kg/d) or a high (21 mg/kg/d) dose of arecoline by gavage. At the termination of the treatments, serum total cholesterol and NO levels were measured, and the aorta morphology was analyzed by hematoxylin and eosin staining. The gene expression of monocyte chemoattractant protein-1 (MCP-1) and adhesion molecules in the thoracic aortas was determined by RT-PCR, and the MCP-1 protein expression and NF-κB activity were detected by Western blot analysis. NO production, MCP-1 secretion in cultured rat aortic endothelial cells (RAECs), and monocyte-endothelium adhesion assay were also performed after arecoline treatments.</p><p><b>RESULTS</b>Arecoline efficiently decreased atherosclerotic plaque areas, increased serum nitric oxide (NO) content, suppressed the mRNA and protein expression of MCP-1, and modulated the IκB-α degradation and P65 phosphorylation in the aortae of ApoE-/- mice. Furthermore, arecoline promoted NO production and suppressed MCP-1 secretion in cultured RAECs after ox-LDL exposure, and either atropine or NG-nitro-L-arginine methylester could abrogate these effects. Arecoline also significantly inhibited the adherence of U937 monocytes to the ox-LDL injured human umbilical vein endothelial cells, which could be abolished by atropine.</p><p><b>CONCLUSION</b>Our results indicate that arecoline attenuates the progression of atherosclerosis and inhibits endothelial cells activation and adherence by stimulating endothelial NNMR. These effects, at least in part, are due to its modulation on NF-κB activity.</p>

The role of arecoline on hepatic insulin resistance in type 2 diabetes rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effects of arecoline on hepatic insulin resistance in type 2 diabetes rats and to elucidate its possible mechanism.</p><p><b>METHODS</b>Forty five Wistar rats were fed with high fructose diet for 12 weeks to induce type 2 diabetic rat model. rats were randomly divided into 5 groups (n = 8): control group, model group and model group were treated with different dose (0, 0.5, 1, 5 mg/kg) of arecoline. After 4 weeks, the fasting blood glucose, blood lipid and insulin level measured , mRNA expression of liver constitutive androstane receptor (CAR), pregnane X receptor (PXR), glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by reverse transcription polymerase chain reaction (RT-PCR), the protein expression of p-AKT and glucose transporter4 (GLUT4) were detected by Western blot.</p><p><b>RESULTS</b>1.5 mg/kg arecoline could significantly decrease the level of fasting blood glucose, blood lipid, blood insulin level and liver G6Pase, PEPCK, IL-6, TNF-alpha mRNA level in type 2 diabetes rats. 1.5 mg/kg arecoline also could significantly increase CAR, PXR mRNA level and p-AKT and GLUT4 protein expression.</p><p><b>CONCLUSION</b>Arecoline improved hepatic insulin resistance in type 2 diabetes rats by increasing the mRNA levels of CAR and PXR leading to the creased glucose metabolism and inflammation related genes expression.</p>

The Hepatotoxicity and Testicular Toxicity Induced by Arecoline in Mice and Protective Effects of Vitamins C and E

Arecoline is a major alkaloid of areca nuts which are widely chewed by southeast Asian and it manifests various toxic effects in different organs of human and animals. In this work, mature mice were treated by vitamins C plus E, arecoline, or both daily for four weeks. The results showed that arecoline significantly increased the levels of serum alkaline phosphatase (ALP), glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and significantly decreased the levels of reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) in the liver tissues. Additionally, the body weight, testis weight, sperm counts, motility and normal sperms also were significantly decreased. The supplement of vitamins C and E can bring the activities of ALP and GPT to normal levels and partially restore the sperm counts compared to the arecoline-treated group but have no other positive effects. In conclusion, the vitamins C and E partially attenuated the arecoline-induced hepatotoxiciy but basically had on protective effects against the arecoline-induced testicular toxicity.

Constituents Released from Streptococcus mutans Attenuate Arecoline-mediated Cytotoxicity in HGF Cells by Altering Intracellular Ca2+ Signaling

Streptococcus mutans (S. mutans) is a facultative anaerobic bacterium mainly found in the oral cavity and is known to contribute to tooth decay and gingivitis. Recent studies on intestinal microbiota have revealed that microorganisms forming a biofilm play important roles in maintaining tissue homeostasis through their own metabolism. However, the physiological roles of oral microorganisms such as S. mutans are still unclear. In our current study, we identified that constituents released from S. mutans (CR) reduce arecoline-mediated cytotoxicity without producing toxic effects themselves. Arecoline, as a major alkaloid of areca nut, is known to mediate cytotoxicity on oral epithelial cells and induces a sustained intracellular Ca2+ ([Ca2+]i) increase that is cytotoxic. The exposure of human gingival fibroblast (HGF) cells to CR not only inhibited the sustained [Ca2+]i increase but also the initial [Ca2+]i elevation. In contrast, CR had no effects on the gene regulation mediated by arecoline. These results demonstrate that S. mutans has physiological role in reducing cytotoxicity in HGF cells and may be considered a novel pharmaceutical candidate.

Non-neuronal muscarinic receptor activation prevents apoptosis of endothelial cells induced by homocysteine / 中国应用生理学杂志

<p><b>OBJECTIVE</b>Endothelial apoptosis plays an important role in the initiation of atherosclerosis. It would be useful to clarify whether activation of non-neuronal muscarinic receptor (NNMR) could prevent endothelial apoptosis and atherosclerosis. We investigated the effects of NNMR activation on regulating rat aortic endothelial cells (RAECs) apoptosis induced by homocysteine, an independent risk factor of atherosclerosis, and further studied its molecular mechanism.</p><p><b>METHODS</b>RAECs were incubated using homocysteine at the concentration of 2.7 mmol/L for 36 h. RAECs were also pre-treated with carbachol or arecoline to examine their effects. RT-PCR was used to assess changes in the gene expression related to cell apoptosis.</p><p><b>RESULTS</b>Incubation of RAECs with homocysteine at the concentration of 2.7 mmol/L resulted in morphologic changes, such as cellular shrinkage, membrane blebbing, chromatin condensation and margination. These could be attenuated by pretreatment with carbachol and arecoline at the concentration of 10 micromol/L for 12 h. Homocysteine induced apoptosis in RAECs and the molecular mechanisms were associated with the regulation of fas, fas-L and caspase-8 in the death receptor pathway, bcl-2, bcl-xL and bax in the mitochondrial pathway, caspase-12 in the endoplasmic reticulum pathway and caspase-3, caspase-6 and p53 as downstream effectors. Carbachol and arecoline attenuated the effects of homocysteine on genes in the death receptor pathway, in the mitochondrial pathway and in the downstream pathway. Atropine could reverse all of the effects of arecoline.</p><p><b>CONCLUSION</b>Activation of NNMR by carbacol and arecoline inhibits homocysteine-induced endothelial cell apoptosis mainly through regulation of death receptor pathway, mitochondrial pathway and downstream effectors.</p>

Efectos bucales de la nuez de betel / Oral effects of the betel nut

La nuez de betel o nuez de areca es una semilla de la palmetra de betel (areca catechu), una de las plantas más polulares del mundo. Diversos pueblos asiáticos, por influencias culturales, tienen la costumbre de masticar la semilla de esta especie vegetal. Entre sus principios activos, la arecaina y la arecolina, son alcaloides comparables a la nicotina en los efectos nerviosos estimulantes. La sustancia activadora causa un relajamiento agradable en la boca, sensación que se propaga al sistema nervioso central. Sin embargo, masticar regularmente la nuez de betel, tiñe la saliva de rojo vivo y ennegrece los dientes, siendo extremadamente perjudicial para la salud oral, causando la pérdida precoz de dientes. A pesar de sus efectos maléficos, existe la dificultad de erradicar el hábito, debido a su carácter cultural, donde las manchas son motivo de orgullo. La agencia internacional de investigación del cáncer (IARC) clasifica a la nuez de betel como un cancerígeno, existiendo numerosos estudios que relacionen la costumbre con neoplasias bucales. El estudio tiene por finalidad realizar un análisis crítico sobre el uso de esta sustancia, buscando informar a la comunidad para prevenir sus efectos maléficos.

The betel nut or areca nut is the seed of the palm tree of bétele (areca catechu), one of the most popular plants of the world. Asians, by cultural influence, have the custom of chew the seed of this sort of vegetable. Between its active principles: arecaine and arecoline, they are alkaloids comparable to the nicotine in the neural stimulation effects. The active substance causes a pleasant relaxion sensation in the mouth, sensation that is extended to the central nervous system. However, chewing regularly betel nut dryes of red the saliva and blackens the teeth being extremely harmful to the mouth health causing early the loss of the teeth. Despite of the harmful effects, the difficulty to eliminate this habit is due to its deep cultural character. Chewing bétel nuts and have the teeth stained it's a motive of proud. The International Agency of Research of the Cancer (IARC) classifies the bétel nut as carcinogenic acquaintance, and the literature have many studies that relate this custom with the oral cancer. The study has as goal to do a critical analysis of the utilization of this substance, searching to inform to the community their bad effects.

Frequency- and state-dependent blockade of human ether-a-go-go-related gene K+ channel by arecoline hydrobromide / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The rapidly activating delayed rectifier potassium current (I(Kr)), whose pore-forming alpha subunit is encoded by the human ether-a-go-go-related gene (hERG), is a key contributor to the third phase of action potential repolarization. The aim of this study was to investigate the effect and mechanism of arecoline hydrobromide induced inhibition of hERG K(+) current (I(hERG)).</p><p><b>METHODS</b>Transient transfection of hERG channel cDNA plasmid pcDNA3.1 into the cultured HEK293 cells was performed using Lipofectamine. A standard whole-cell patch-clamp technique was used to record the I(hERG) before and after the exposure to arecoline.</p><p><b>RESULTS</b>Arecoline decreased the amplitude and the density of the I(hERG) in a concentration-dependent manner (IC(50) = 9.55 mmol/L). At test potential of +60 mV, the magnitude of I(hERG) tail at test pulse of -40 mV was reduced from (151.7 ± 6.2) pA/pF to (84.4 ± 7.6) pA/pF (P < 0.01, n = 20) and the magnitude of I(hERG) tail at test pulse of -110 mV was reduced from (-187.5 ± 9.8) pA/pF to (-97.6 ± 12.6) pA/pF (P < 0.01, n = 20). The blockade of arecoline in the open and inactivated state was significant in a state-dependent manner. The maximal blockade was achieved in the inactivated state. Studies of gating mechanism showed that the steady-state activation curve of I(hERG) was significantly negatively shifted by arecoline. Time constants of activation were shortened. Steady-state inactivation curve and time constants of fast inactivation were not significantly affected by arecoline. Furthermore, the inhibition of I(hERG) by arecoline was characterized markedly by a frequency-dependent manner from 0.03 to 1.00 Hz pulse.</p><p><b>CONCLUSION</b>Arecoline could potently block I(hERG) in both frequency and state-dependent manner.</p>

Determination of synephrine and arecoline contents in Xiao'er Xiaoji Zhike oral liquid / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a HPLC method for the determination of synephrine and arecoline contents in Xiao'er Xiaoji Zhike oral liquid.</p><p><b>METHOD</b>The analysis was performed on a Symmetry C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-methanol-0.1% acetic acid solution (containing potassium dihydrogen phosphate 0.6 g and SDS 1.0 g per 1 000 mL) (15: 30: 55) as mobile phase. The detection wavelength was set at 215 nm.</p><p><b>RESULT</b>The synephrine and arecoline concentrations had good linear relationship between 0.281 5-2.815 and 0.040 16-0.401 6 microg (r > 0.999 7), and the average recoveries of the two compounds were 97.1% (RSD 1.4%) and 100% (RSD 1.3%), respectively.</p><p><b>CONCLUSION</b>The method is simple, accurate and sensitive.</p>

Oral submucous fibrosis : current concepts in etiopathogenesis.

Oral submucous fibrosis (OSF) is now accepted globally as an Indian disease, having highest malignant potential than any other oral premalignant lesions. The understanding of the exact role of alkaloids and other etiological agents with respect to pathogenesis will help the management and minimize the blind clinical trials and treatment modalities. This article provides an overview of the etiopathogenesis with stress on the recent concepts related to this chronic “Indian Disease”.

Effects of arecoline and nicotine on the expression of hTERT in oral keratinocytes / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of arecoline and nicotine on the expression of human telomerase reverse transcriptase (hTERT) mRNA and protein in cultured normal human oral keratinocytes (KC).</p><p><b>METHODS</b>The experiments were divided into arecoline group, arecoline/nicotine group and control group. The hTERT mRNA and protein expression of KC was examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot.</p><p><b>RESULTS</b>Arecoline could induce the hTERT mRNA and protein expression of KC in a dose dependent manner, the hTERT mRNA and protein expression of KC was higher in 0.030, 0.060, 0.090 g/L arecoline group than control group (P < 0.001). Nicotine (0.025 g/L) increased hTERT mRNA and protein expression of KC induced by arecoline.</p><p><b>CONCLUSIONS</b>Arecoline could increase the expression of hTERT mRNA and protein in oral keratinocytes. Nicotine had a synergistic effect on arecoline. hTERT over-expression induced by arecoline and nicotine may play an important role in the malignant transformation of oral submucous fibrosis.</p>

Effect of arecoline on the differentiation of myofibroblasts of oral mucosa / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the origin of myofibroblasts in oral submucous fibrosis.</p><p><b>METHODS</b>The oral keratinocytes and fibroblasts were isolated and cultured. The expression of the alpha-smooth muscle actin in the fibroblasts was examined by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>No difference was found in expression of alpha-smooth muscle actin between the fibroblasts that were directly stimulated by arecoline and the control. The expression of alpha-smooth muscle actin in the keratinocyte and fibroblast-cocultured group was higher than in the control group, and higher in fibroblasts cocultured with keratinocytes preprocessed by arecoline than in fibroblasts cocultured with keratinocytes without preprocessed by arecoline.</p><p><b>CONCLUSIONS</b>The differentiation of myofibroblasts from fibroblasts in oral submucous fibrosis might be induced by the interaction of arecoline and keratinocyte.</p>

Vigilancia epidemiologica de la equinococcosis quistica en perros, establecimientos ganaderos y poblaciones humanas en la Provincia de Rio Negro / Vigilancia epidemiologica de la equinococcosis quistica en perros, establecimientos ganaderos y poblaciones humanas en la Provincia de Rio Negro

La vigilancia de la equinococcosis quística para detectar infestación por Echinococcus granulosus en la Provincia de Río Negro en el período 1980-2002 fue efectuada en el hombre mediante encuestas serológicas y ultrasonográficas en población joven, y en el perro por el test de arecolina. Dadas las limitaciones de esta técnica, se planteó suplantarla por el complejo copro ELISA Western Blot en heces caninas recolectadas del suelo. El objetivo del presente trabajo fue comparar las ventajas y limitaciones de las dos técnicas para medir la prevalencia de la infección en el perro, y evaluar la prevalencia actual de la infección en el hombre y en el perro. Elárea de trabajo comprendió 7 Departamentos endémicos con Programas de desparasitación canina sistemática (Area Programa) y 4 Departamentos no endémicos como Area Testigo. El test de arecolina se aplicó en los perros, con concurrencia voluntaria de sus propietarios (muestreo no aleatorizado). Las muestras para detección de coproantígenosfueron obtenidas de establecimientos ganaderos seleccionados en forma aleatorizada. En el hombre se determinó la prevalencia mediante tamizajes ultrasonográficos en escolares de 6 a 14 años y la incidencia por medio del sistema oficial de notificación de casos sintomáticos. Se dosificaron con arecolina 416 perros resultando 19 (5.2%) positivos en el Area Programa y ninguno positivo en el Area Testigo. Para la detección de coproantígenos se obtuvieron 748 muestras de materia fecal de 271 establecimientos ganaderos, resultando 37 muestras y 32 establecimientos (13.6%) positivos en el Area Programa y 4 muestras y 4 establecimientos (11.4%, IC: 0.3-32.3) positivos en el Área Testigo. En el Area Programa se efectuaron 7421 ecografías abdominales a escolares, detectándose 40 (0.5%) casos conimágenes compatibles con hidatidosis, mientras en el área testigo se efectuaron 1732 ecografías con 9 (0.5%) casos positivos...

The surveillance of infection for Echinococcus granulosus in the Provinceof Rio Negro during 1980-2002 included serological and ultrasonographic screening in humans and arecoline testin dogs. In lieu of the limitations of the arecoline test the proposal was to supplant that test for the copro Elisacopro/Western Blot complex applied to feces collected from the environment. The objective was to compare the pros and cons of the two tests and to evaluate the human and the canine infection prevalence. The working area encompassed 7 Departments with systematic canine parasiticide activities (Program Area) and 4 Departments, not endemic, as Control Area. The arecoline test was applied to the dogs in assembled groups with the voluntary participation of their owners (not randomized sampling). Samples for the detection of coproantigens were obtained from sheep farms selected at random and analyzed by the complex copro-LISA /Western Blot. Prevalence inman was determined by screening the school population (6 to 14 years old) by ultrasound, and by means of the compulsory notification of cases from the official system. Dogs (416) were tested with arecoline, 365 of whichbelonged to the Program Area. Of these 19 (5.2%) resulted positive, while none of 51 dogs from the Control Areawere positive. Samples (748) of feces were tested to detect coproantigens, obtaining 37 positive samples withinthe Program Area and 4 within the Control Area. Farms (271) from the livestock estate unit were evaluated, outof which 236 belonged to the Program Area, gave 32 (13.6%) positive results, while 4 (11.4%) of 35 from theControl Area resulted positive. Sonography tests (7421) were done in the Program Area detecting 40 (0.5%)carriers, while in the Control Area, over 1732 tests, 9 (0.5%) resulted positive...

Synergic effects of synthesis arecoline in combination with snail-killing drugs niclosamide / 中华预防医学杂志

<p><b>OBJECTIVE</b>To prove that synthetic Are combination with snail-killing drug Nic can increase the effects of snail-killing remarkably.</p><p><b>METHODS</b>In indoor immersing experimentation, the experiments were divided into 4 groups, 30 snails in each group, to observe the rate of opening operculum, the rate of climbing adhesion and the rate of death at 3, 6 and 24 hours respectively. In field experimentation, we intermixed 0.1 mg/L Are with 0.2 mg/L Nic as sample as contrasted with 2 mg/L Nic and non-drug group. Immersing method (we chose three slots each size were 10 m x 2 m x 1 m.) and insufflation method (we chose three patch of bottomlands each area were 10 m x 5 m.) were used to kill snails separately and the death rate of fish, at the same time was observed.</p><p><b>RESULTS</b>In the room, as we added 0.1 mg/L Are to the solution of 0.1 mg/L and 0.2 mg/L Nic separately, the opening operculum rate for 6 hours was increased from 20% and 12% to 100% and 95%, the climbing adhesion rate for 6 hours decreased from 17% and 53% to 3% and 5%, the death rate for 24 hours increased from 25% and 40% to 90% and 100%. In the field, the snails death rate in sample group and in contrastive group applied with immersing method and insufflation method for 72 hours were 95.9%, 93.3% and 100%, 95.8%; only one small fish (2 cm long) died in sample group, and all fishes died in Nic group, and all fish were alive in non-drug group.</p><p><b>CONCLUSION</b>It proved that synthetic Are combination with snail-killing drug Nic might decrease Nic dosage and toxicity and increase the effects of snail-killing.</p>