Vasculite urticariforme e exantema: uma reação de hipersensibilidade tardia mista a dimenidrinato ­ relato de caso / Urticarial vasculitis and rash: a mixed delayed hypersensitivity reaction to dimenhydrinate ­ case report

Dimenidrinato é um anti-histamínico H1 do grupo das etanolaminas, com importantes propriedades anticolinérgicas, antisserotoninérgicas e sedativas. Relatamos um caso de uma mulher que após 14 dias de ter usado dimenidrinato, iniciou quadro de exantema e vasculite urticariforme, além de sintomas constitucionais. Avaliação laboratorial sem alterações. Biopsia de pele evidenciou dermatite de interface do tipo vacuolar e púrpura com leucocitoclasia e derrame pigmentar. Imunofluorescência positiva para IgG, com presença de fluorescência dos núcleos dos queratinócitos da epiderme. Tratada com corticoide oral por 2 meses até remissão completa do quadro, e posterior realização de teste intradérmico, que foi positivo na leitura de 48h. A reação de hipersensibilidade tardia observada foi relacionada a mecanismo misto de Gell e Coombs (III e IV), com positividade no teste cutâneo intradérmico de leitura tardia em 48h (reação tipo IV) e biópsia compatível com vasculite cutânea (reação tipo III); lesões exantemáticas (reação tipo IV) e urticária vasculítica (reação tipo III). O teste cutâneo com dimenidrinato positivo reforça o diagnóstico de reação de hipersensibilidade.

Dimenhydrinate is an H1 antihistamine from the ethanolamine group, with important anticholinergic, antiserotoninergic and sedative properties. We report the case of a woman who, after 14 days of using dimenhydrinate, developed rash and urticarial vasculitis, in addition to constitutional symptoms. Laboratory tests were normal. Skin biopsy revealed interface purpuric dermatitis with leukocytoclasia and pigment effusion. Immunofluorescence was positive for IgG, showing nuclear fluorescence of epidermal keratinocytes. She was treated with oral corticosteroid for 2 months until complete remission of symptoms. Subsequent intradermal test resulted positive on the 48-h reading. The delayed hypersensitivity reaction was related to a mixed Gell and Coombs mechanism (III and IV), with positive results in the intradermal cutaneous test on the 48-h reading (type IV reaction) and a biopsy compatible with cutaneous vasculitis (type III reaction), exanthematous lesions (type IV reaction,) and urticarial vasculitis (type III reaction). The positive skin test for dimenhydrinate reinforces the diagnosis of hypersensitivity reaction.

Anti-Oxidative Effect of Myrtenal in Prevention and Treatment of Colon Cancer Induced by 1, 2-Dimethyl Hydrazine (DMH) in Experimental Animals

Colon cancer is considered as the precarious forms of cancer in many developed countries, with few to no symptoms; the tumor is often diagnosed in the later stages of cancer. Monoterpenes are a major part of plant essential oils found largely in fruits, vegetables and herbs. The cellular and molecular activities show therapeutic progression that may reduce the risk of developing cancer by modulating the factors responsible for colon carcinogenesis. Colon cancer was induced with DMH with a dose of (20 mg/Kg/body weight) for 15 weeks by subcutaneous injection once in a week. Myrtenal treatment was started with (230 mg/Kg/body weight) by intragastric administration, one week prior to DMH induction and continued till the experimental period of 30 weeks. The Invivo results exhibit the elevated antioxidant and lipid peroxidation levels in DMH treated animals. The Histopathological analysis of colon tissues well supported the biochemical alterations and inevitably proves the protective role of Myrtenal. Treatment with myrtenal to cancer bearing animals resulted in a remarkable increase in the inherent antioxidants and excellent modulation in the morphological and physiological nature of the colon tissue. It is thus concluded that myrtenal exhibits excellent free radical scavenging activity and anticancer activity through the suppression of colon carcinoma in Wistar albino rats.

Dexametasona versus dimenhidrinato en la profilaxis de las náuseas y vómitos postoperatorios en pacientes adultos sometidos a cirugía general y digestiva / Dexamethasone versus dimenhydrinate for prophylaxis of postoperative nausea and vomiting in adult patients undergoing general and digestive surgery

Introducción. Las náuseas y vómitos postoperatorios (NVPO) son complicaciones frecuentes en los pacientes quirúrgicos. Se han publicado una serie de guías clínicas y revisiones que recomiendan diferentes fármacos para la profilaxis y tratamiento de NVPO. Los fármacos dexametasona (DEX) y dimenhidrinato (DIM) son ampliamente utilizados para este fin; sin embargo, la evidencia científica que apoya la efectividad del DIM en el contexto de NVPO, es escasa. Objetivo. Determinar la dosis del fármaco (DIM o DEX con mayor efectividad en la profilaxis de náuseas y vómitos postoperatorios en pacientes adultos sometidos a cirugía general y laparoscópica. Asimismo, determinar la aparición de náuseas, vómitos, náuseas y vómitos postoperatorios, necesidad de tratamiento de rescate y efectos adversos de ambos medicamentos. Material y Método. Ensayo clínico aleatorizado realizado en el Hospital II EsSalud de Talara, Piura, Perú con 102 participantes (18 hombres y 84 mujeres) con un riesgo bajo y moderado para NVPO (09 y 93 respectivamente), los que fueron asignados en dos grupos de 51 pacientes cada uno. Un grupo de pacientes recibió DEX (4 mg) y otro DIM (50 mg) luego de la inducción de la anestesia general. Resultados. La incidencia de NVPO en la población tratada con DEX fue de 7,84% y de 39,22% en la población de pacientes que recibieron DIM. Conclusiones. La administración de 4 mg de dexametasona en el acto anestésico provee mejor profilaxis de náuseas y vómitos postoperatorios respecto a 50 mg de dimenhidrinato.

Background. Postoperative nausea and vomiting (PONV) are common complications in surgical patients. There have been a number of clinical guidelines and reviews that recommend different drugs for prophylaxis and treatment of PONV. Dexamethasone and dimenhydrinate drugs are widely used in our country for this purpose, but the scientific evidence supporting the effectiveness of dimenhydrinate in the context of PONV, is scarce. Objective. To determine the dose of the drug (dimenhydrinate or dexamethasone) more effective in the prophylaxis of postoperative nausea and vomiting in adult patients undergoing general and laparoscopic surgery. Also determine the occurrence of nausea, vomiting, PONV, need for rescue treatment and adverse effects of both drugs. Material and Method. Clinic randomized trial performed in the Hospital II Essalud Talara, Piura, Peru, with 102 participants (18 men and 84 women) with a low or moderate risk for PONV (09 and 93 respectively), which were assigned into two groups of 51 patients each. A group of patients received dexamethasone (4 mg) and another dimenhydrinate (50 mg) after induction of general anesthesia. Results. The incidence of PONV in the dexamethasonetreated population was 7,84% and 39,22% in the population of patients who received dimenhydrinate. Conclusions. The dexamethasone administration of 4 mg during anesthesia provides better prophylaxis of postoperative nausea and vomiting over dimenhydrinate administration of 50 mg.

Medicamentos antieméticos no tratamento da náusea e vômitos associados à gestação / Antiemetic medications in the treatment of nausea and vomiting associated with pregnancy

Náusea e vômitos (NV) são os sintomas mais comuns durante a gravidez, geralmente iniciando-se entre a 6ª e a 8ª semana, atingindo a intensidade máxima em torno da 9ª semana e resolvendo-se até a 12ª semana. Embora sua etiologia seja, provavelmente, multifatorial, seu curso clínico se correlaciona com as concentrações plasmáticas da gonadotrofina coriônica humana. Por comprometerem a qualidade de vida, NV devem ser abordados com modificações dietéticas que incluem dieta fracionada e redução do consumo de alimentos gordurosos, entre outras. O uso de piridoxina pode melhorar a náusea de intensidade leve, embora não diminua significantemente os episódios de vômitos. Os anti-histamínicos são os medicamentos mais utilizados como terapia medicamentosa de primeira linha e têm sua segurança comprovada; dentre eles, o dimenidrinato determina início de ação mais rápido e menor sedação que a meclizina. Entre os antagonistas dopaminérgicos, a prometazina e a metoclopramida podem ser utilizadas, mas apresentam como desvantagem o potencial de eventos adversos maternos. O antagonista dos receptores 5-HT3, ondansetrona, pode ser considerado quando outros medicamentos não foram efetivos no tratamento de NV de intensidade grave. Do mesmo modo, os corticosteroides devem ter seu uso reservado para casos não responsivos a outros medicamentos e preferencialmente após a 10ª semana de gestação...

Comparison of the effectiveness of intravenous piracetam and intravenous dimenhydrinate in the treatment of acute peripheral vertigo in the emergency department

<p><b>INTRODUCTION</b>We aimed to compare the effectiveness of intravenous piracetam with that of intravenous dimenhydrinate in the treatment of acute peripheral vertigo in the emergency department.</p><p><b>METHODS</b>This double-blind study comprised a total of 200 patients, aged between 18 and 70 years, who had presented to the emergency department of Ankara Training and Research Hospital and were diagnosed with peripheral vertigo. Evaluation of the severity of the patients' vertigo was performed using a visual analogue scale, before and after drug administration.</p><p><b>RESULTS</b>Both drugs were found to be effective (p < 0.001) and had comparable effects (p < 0.474). Dimenhydrinate was also found to have about two times the side effects of piracetam. Drowsiness was found to be the most common side effect of these two drugs.</p><p><b>CONCLUSION</b>Dimenhydrinate and piracetam have similar levels of effectiveness with regard to acute vertigo. We conclude that piracetam, which has fewer side effects than dimenhydrinate, better vestibular compensation, and is effective for both acute and chronic vertigo, could be more frequently used in the emergency treatment of acute vertigo.</p>

Sequential Changes in Aberrant Crypt Foci and Lectin Expression in the Early and Late Stages of DMH-Induced Colon Carcinogenesis in Rats

BACKGROUND/AIMS: The purpose of this study was to investigate the malignant potential of aberrant crypt foci (ACF) by measuring the multiplicity of crypts and lectin expression in the early and late stages of 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis. METHODS: Six-week-old Wistar rats were injected subcutaneously with DMH for 27 weeks. We classified ACF according to the number of crypts per ACF as a few crypts ( or =4 crypts, NC ACF). Immunohistochemistry was used to evaluate lectin expression. RESULTS: In the early stage, FC ACF (590/1,902, 31.0%) occurred more frequently than NC ACF (35/449, 7.8%); whereas in the late stage, NC ACF (176/449, 39.2%) occurred more frequently than FC ACF (324/1,902, 17.0%). The number of ACF peaked at 15 to 20 weeks. The ratio of NC/FC ACF increased gradually during carcinogenesis. The expression of both UEA1 and PNA was higher in NC ACF than FC ACF. Lectin expression increased in the late stage compared with the early stage. CONCLUSIONS: The expression of lectin was higher in NC ACF and ACF in the late stage. Therefore, ACF with higher multiplicities in the late stage may have more malignant potential in DMH-induced colon carcinogenesis.

Physicochemical Requirement for the Vegetative Growth of Schizophyllum commune Collected from Different Ecological Origins

Schizophyllum commune is an edible and medicinal mushroom widely distributed in the world. The optimal growth conditions for the mycelia of 10 strains of the fungus were investigated. The temperature suitable for the mycelial growth and density was obtained at 30~35degrees C. Among the tested conditions, the minimum mycelial growth was found at 15degrees C. In case of pH, the most favorable growth was found at pH 5. The results indicated that this mushroom well adapted to high temperature and low pH for its mycelial growth. Considering growth phenotype of mycelia, Hamada, Hennerberg, PDA and YM were the most suitable and Lilly, Glucose triptone, Glucose peptone and Hoppkins were the most unfavorable among tested media for the mycelial growth of S. commune. Out of tested carbon sources, dextrin and fructose were the most suitable and lactose, mannose and sorbitol were the unsuitable for the fungus. Compact mycelial density was obtained from most of the carbon sources. Among used nitrogen sources, calcium nitrate, potassium nitrate and alanine were the most appropriate and the most incompatible were ammonium phosphate, histidine, urea and arginine for mycelial growth of S. commune on the culture media. Calcium nitrate, histidine and potassium nitrate showed moderately thin or thin, and rest of nitrogen sources showed compact or moderately compact mycelial density.

Role of metochlopramide and dimenhydrinate in orevention of postoperative nausea and vomiting in laparoscopic gynaecological surgery

To evaluate the role of metoclopramide and dimenhydrinate in controlling postoperative nausea and vomiting [PONV] and its cost effectiveness in gynaecological laparoscopy. This study was conducted in the department of anaesthesiology and intensive care unit, Pakistan Institute of Medical Sciences, Islamabad from June 2004 to March 2006. Ninety nine female patients belonging to American society of Anaesthesiologist [ASA] grading ASA-1 to ASA-111, scheduled for laparoscopic surgery, who did not fall in exclusion criteria were finally included. Anaesthetic technique was standardized for all patients. Injection Metocloparamide 10 mg and injection Dimenhydrinate 50 mg were administered 20 min before the procedure was over. At the end of procedure patients were transferred to the recovery room for observation for 10 hours. Four point verbal descriptive scale [VDS] was used to identify the presence and severity of PONV. Four out of 99 [4.04%] patients developed nausea soon after regaining consciousness and did not demand any medication for relief. Three [3.03%] patients developed vomiting. It was single episode and no rescue medication was needed. Most of the symptoms developed with in 10 -30 minutes of reversal. Metocloparamide and dimenhydrinate is a good combination to combat PONV and is costeffective in laparoscopic gynaecological surgery

A randomized comparison of ginger and dimenhydrinate in the treatment of nausea and vomiting in pregnancy.

OBJECTIVE: To study the efficacy of ginger and dimenhydrinate in the treatment of nausea and vomiting in pregnancy. STUDY DESIGN: Double blind randomized controlled trial. SETTING: Department of Obstetrics and Gynecology, Thammasat Hospital, Faculty of Medicine, Thammasat University. MATERIAL AND METHOD: Between January 2005 and December 2005, 170 pregnant women who attended at antenatal clinic Thammasat University Hospital with the symptoms of nausea and vomiting in pregnancy were randomly allocated into group A (n = 85) and group B (n = 85). The patients in group A received one capsule of ginger twice daily (one capsule contained 0.5 gm of ginger powder) while the patients in group B received the identical capsule of 50 mg dimenhydrinate twice daily. The visual analogue nausea scores (VANS) and vomiting times were evaluated at day 0-7 of the treatment. RESULTS: There was no significant difference in the visual analogue nausea scores (VANS) between group A and group B in day 1-7 of the treatment. The vomiting episodes of group A were greater than group B during the first and second day of the treatment with statistically significant difference. No difference in vomiting episodes during the day 3-7 of treatment was found in both groups. There was a statistically significant difference in the side effect of drowsiness after treatment in group B greater (77.64%) than group A (5.88%) (p < 0.01). CONCLUSION: From the presented data, ginger is as effective as dimenhydrinate in the treatment of nausea and vomiting during pregnancy and has fewer side effects.

Characterization of the Expression of PKCalpha(Isoform) in DMH-induced Vascular Endothelial Proliferation

PURPOSE: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of PKCalpha, and mu was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of PKCalpha on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. PKCmu will be investigated in further study. METHODS: The study was conducted with the cultured HUVECs group(control) and the 0.75x10(-9)M DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of PKCalpha was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular PKCalpha particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. RESULTS: The Western blot analysis showed increased PKCalpha expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. CONCLUSION: We believe that PKCalpha plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

Effect of Antivertigo Medications on Vestibular Function in Healthy Human Subjects / 항공우주의학회지

BACKGROUND: Motion sickness is one of the major problems of aerospace medical concern. Vestibule plays an important role in giving rise to motion sickness. Drugs preventing motion sickness have a suppressive effect on the vestibular function through the antagonistic effect to some receptors in vesibular nuclei and vomiting center of central nervous system. We identified and quantified the effects of anti-motion sickness drugs on vestibule-ocular reflex in healthy human subjects. METHODS: Fourty-five healthy male subjects were grouped to one of placebo, dimenhydrinate 50 mg, scopolamine (1 patch), or both scopolamine and dimenhydrinate group, and received rotation chair test before and after drug administration to obtain Vestibulo-ocular reflex (VOR) gain and phase in sinusoidal harmonic acceleration (SHA) with frequencies of 0.01, 0.02, 0.04 and 0.08 Hz. The delta gain and the delta phase by the drug administration were obtained and analyzed as pharmacodynamic effects. RESULTS: Baseline gain and phase data were not different by the groups in all SHA frequencies. VOR gains were significantly decreased by 0.15~0.17 after dimenhydrinate administration. In the scopolamine group, there were significant decreases in 0.04 and 0.08 Hz by 0.14 and 0.15, respectively, but no difference in 0.01 and 0.02 Hz was observed. Increasing tendency in VOR phase lead was observed, especially in dimenhydrinate, but not significantly. There was no additive effect on the reduction of VOR gain when the two drugs were co-administered. CONCLUSION: We quantitatively characterized how much the VOR parameters were changed by the drugs with different kinds of mechanism. Dimenhydrinate reduced the VOR gain by around 0.16. However, scopolamine probably has a minimal or no additive effect on VOR suppression.

Role of Protein Kinase C in Abnormal Proliferation of Vascular Endothelial Cell induced by 1,2-Dimethylhydrazine; Analysis of Isoform

PURPOSE: Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, have been known to take a role in signal transduction pathway of angiogenesis. The authors confirmed that PKC is the most noticeable mediator for abnormal proliferation of vascular endothelial cells through in vitro study model using the inhibitors, targeting the formation of three co-enzymes. In this study, we would investigate which isoform of PKC play an important role in abnormal angiogenesis of vascular endothelial cell. METHODS: In 96 well plates, 10(4) HUVECs(human umbilical vein endothelial cells) were evenly distributed. Two groups were established; the control group without administration of DMH(1,2-dimethylhydrazine) and the DMH group with administration of 7.5x10(-9)M DMH. RNA was extracted from vascular endothelial cell of each group and expression of the PKC isoform was analyzed by RT-PCR(reverse transcriptase-polymerase chain reaction) method. RESULTS: RT-PCR analysis showed that PKCalpha, -betaI, -betaII, -eta, -micron and -zeta were expressed in vascular endothelial cells of each group. DMH incresed the expression of PKCalpha and PKCmicron, and decreased PKCbetaI, PKCbetaII expression dominantly. CONCLUSION: Based on the result of this study, it was suggested that PKCalpha and PKCmicron may have significant role in abnormal proliferation of vascular endothelial cell.

Activity of Protein Kinase C in Abnormally Proliferated Vascular Endothelial Cells

PURPOSE: To understand the pathogenesis of the disease that presents abnormally proliferated vascular endothelial cells, a model of DMH(1,2-dimethylhydrazine)-induced abnormal proliferation of HUVECs(Human Umbilical Vein Endothelial Cells) was made. We indirectly determined that Protein Kinase C(PKC) restricts the cellular proliferation and inhibits the manifestation of growth factor by using several inhibiting substances of the transmitter through our previous studies. Thereupon, we attempted to observe direct enzymatic activities of PKC and its correlation with the abnormal proliferation of vascular endothelial cells. METHODS: 10(5) HUVECs cells were applied to 6 individual well plates in three different groups; A control group cultured without treatment, a group concentrated with 0.75x10(-8)M DMH only, and a group treated with DMH & 5x10(-9)M Calphostin C, inhibitor of PKC. In analyzing the formation of intracellular PKC enzyme, protein separation was performed, and separated protein was quantitatively measured. PKC enzyme reaction was analyzed through Protein Kinase C Assay System (Promega, USA), and the results were analyzed according to Beer's law. RESULTS: Enzymatic activity of PKC presented the highest in all reaction time of a group concentrated only with DMH, and the lowest in the control group. The group treated with DMH and the inhibitor revealed statistically lower enzymatic activity than group only with DMH in all reaction time, although higher than the control group. CONCLUSION: From the enzymatic aspect, most active and immediate reaction of the PKC was observed in the group concentrated with DMH only. The group treated with DMH & PKC inhibitor showed meaningful decrease. Accordingly, PKC holds a significant role in DMH-induced abnormal proliferation of vascular endothelial cells.

Contribution to the treatment of nausea and emesis induced by chemotherapy in children and adolescents with osteosarcoma

CONTEXTO E OBJETIVO: A êmese induzida por quimioterapia é fator limitante no tratamento de crianças com câncer. O uso de quimioterapia com drogas emetogênicas tem aumentado a freqüência desse efeito colateral. O objetivo é comparar a eficácia e a toxicidade do granisetron às da combinação de altas doses de metoclopramida e dimenidrato em crianças com osteossarcoma utilizando a mesma quimioterapia. TIPO DE ESTUDO E LOCAL: Aberto, prospectivo, randomizado, realizado no Instituto de Oncologia Pediátrica, Departamento de Pediatria, Universidade Federal de São Paulo, Brasil. MÉTODOS: Entre fevereiro e agosto de 1994, 26 crianças com idade de 7 a 18 anos (média de 14 anos), recebendo quimioterapia para osteossarcoma, entraram no estudo. A quimioterapia consistiu de ciclos repetidos de: A) ifosfamida 2.500 mg/m² + epirrubicina 75 mg/m²; B) ifosfamida 2.500 mg/m² + carboplatina 600 mg/m²; C) carboplatina 600 mg/m² + epirrubicina 75 mg/m². 80 tratamentos quimioterápicos foram avaliados para o controle de náuse e vômito. Os pacientes foram randomizados para receber dose única de granisetron (50 µ/kg) ou metoclopramida (2 mg/kg) mais dimenidrato (5 mg/kg) infundidos por oito horas. Êmese e náusea foram monitoradas por 24 horas por meio de escore de MANE (Morrow Assessment of Nausea and Emesis). Foram utilizados testes de Qui-quadrado, t e Mann Whitney, além da técnica de análise exploratória de dados. RESULTADOS: O granisetron induziu resposta completa em 62,5% dos pacientes submetidos aos tratamentos quimioterápicos comparado a apenas 10% obtidos com a combinação de metoclopramida associado ao dimenidrato (p < 0,0001). CONCLUSÕES: Concluímos que o granisetron é droga segura e eficiente em crianças com osteossarcoma superior à associação de metoclopramida e dimenidrato no controle de náuseas e vômitos induzidos por quimioterapia para osteossarcoma em crianças.

A Study for the Mechanism of Abnormal Proliferation in Vascular Endothelial Cells using Inhibitors to the Signal Transduction Pathway

Protein tyrosine kinase(PTK), protein kinase C(PKC), oxidase, as a mediator, take a significant role in signal transduction pathway of angiogenesis. The authors utilized the inhibitors, targeting the formation of three co-enzyme in signal transduction pathway in order to quantify the suppression of abnormal vascular endothelial cell proliferation induced by DMH, to compare the level suppression in each up-regulated growth factors, CTGF, CYR61, ITGbeta1, FHL2, and to identify the relationship between abnormal cell proliferation and signal transduction pathway. Five groups were established; Control group, Group of DMH, Group of DMH-mixed Herbimycin, inhibitor of protein tyrosine kinase, Group of DMH-mixed Calphostin C, inhibitor of protein kinase C, Group Of Dmh-Mixed 10U Catalase, Inhibitor Of oxidase. The rise of vascular endothelial cell was compared by MTT assay, and four growth factors were analysed with RT-PCR method, at pre-administration, 4, 8, 12, and 24 hours after administration. In comparison of abnormal proliferation of vascular endothelial cell induced by DMH, suppression was noticed in Herbimycin and Calphostin C group, and Calphostin C group revealed higher suppression effect. Nevertheless, Catalase group did not have any suppression. In manifestation of four growth factors, Herbimycin and Calphostin C group presented similar manifestation with control group, except in ITGbeta. Catalse group had similar manifestation with DMH group in all four growth factors. Abnormal proliferation of vascular endothelial cell induced by DMH have a direct relationship with PTK and PKC, more specifically to PKC. Oxidase was confirmed not to have any relevance.

Effect of Different Amount of Dietary n-3 PUFA on Colon Carcinogenesis in DMH-treated Rats

The objective of the study was to observe the effect of n-3 PUFA on cell proliferation and apoptosis by determining mRNA and protein of COX-2 and eicosanoid product and the mRNA and protein of Bu and Bcl-2 related to apoptosis in colon carcinogenesis of 1,2- dimethylhydrazine (DMH)-treated rats. Ninety male Sprague Dawley rats weighing about 170g were divided into 3 groups, control and n-3 PUFA supplemented groups (FO group: 6.2 mmoles n-3 PUFA; 2FO group: 12.4 mmoles n-3 PUFA) and fed experimental diet for 14 weeks. All rats were intramuscularly injected with DMH 15 mg/kg twice a week for 6 weeks to deliver total dose of 180 mg/kg body weight. Compared with the control group, 6.2 mmoles n-3 PUFA significantly reduced the levels of mRNA and protein expression of COX-2 and 2-series eicosanoids (TXB2 and PGE2 and decreased cell proliferation in colonic mucosa. However, high levels of n-3 PUFA supplementation significantly increased the levels of mRNA and protein expression of COX-2, TXB2 and PGE2. and increased cell proliferation which was similar level to that of control group. Compared with the control group, n-3 PUFA, regardless of the amount, significantly increased apoptotic index in colonic mucosa. Western blot and RT-PCR analyses showed that the levels of mRNA and protein expression of Bax were significantly increased by 6.2 mmoles n-3 PUFA, but decreased by 12.4 mmoles n-3 PUFA. The analyses also showed the levels of mRNA and protein expression of Bcl-2 were significantly reduced by 6.2 mmoles n-3 PUFA, but increased by 12.4 mmoles n-3 PUFA. The ratio of Bcl-2/Bax in mRNA and protein was significantly reduced by 6.2 mmoles n-3 PUFA but increased by 12.4 mmoles n-3 PUFA. Overall, these results indicate that n-3 PUFA could be effective in preventing colon carcinogenesis by reducing cell proliferation with lower level of COX-2 and 2-series eicosanoid, and increasing apoptosis by inducing pro-apoptotic gene, Bax and inhibiting anti-apoptotic gene, Bcl-2 in the colonic mucosa of DMH-treated rats. However, high level of n-3 PUFA supplementation could stimulate colon carcinogenesis by increasing cell proliferation and inhibiting apoptosis.

RT-PCR of Up-Regulated Factors in Abnormally Proliferated Vascular Endothelial Cells by 1,2- Dimethylhydrazine

Many studies for verifying angiogenesis have been in progress, especially in the field of abnormal vascular proliferation to explain the pathogenesis and to develop a treatment of several diseases. In our previous experiments, endothelial cell proliferations were induced by DMH stimulation in vitro, and the 177 factors(142 up- regulated and 35 down-regulated factors) were identified. Among the up-regulated factors, 9 substances (EFEMP1, CTGF, CYR61, ITGbeta1, FHL2, SERPINE1, MYC, PTTG1 and MSH6) were selected, which were related to cell proliferation and showed high signal intensities. The RNA was isolated from HUVECs at the time of 0, 6, 12, 24 hours after the DMH treatment, and RNA of control group HUVECs was also isolated. Genetic information of selected molecules was used to make primer for each, and RT-PCR was performed to analyze both groups. In control and treatment groups, each substance presented variety of manifestation degree according to time differences. EFEMP1, CTGF, CYR61, ITGbeta1, FHL2 and MYC were related to abnormal vascular proliferation steadily and SERPINE1, PTTG1 and MSH6 were related secondarily. CTGF was related to both normal and abnormal proliferation, but it played a more significant role in abnormal proliferation from earlier stage. EFEMP1, CYR61, ITGbeta1, FHL2 and MYC were similar to CTGF, although the relation appeared lately. Further study should be performed to analyze the expressions and the interactions of growth factors, which could be utilized in the new therapeutic development.

Effect of Dietary Conjugated Linoleic Acid (CLA) Isomers on Tumor Incidence and the Protein Expression of Cyclooxygenase-2 and Protein Kinase C in Colonic Mucosa of DMH-Treated Rats

This study was designed to compare the anti-carcinogenic effect of conjugated linoleic acid isomers on tumor incidence, cell proliferation and the levels of thromboxane (TX)B2, prostaglandin (PG)E2 and 1,2-diacylglycerol (DAG), and the related enzyme expression of cyclooxygenase (COX)-2 and protein kinase C (PKC) in colonic mucosa of 1,2-dimethylhydrazine (DMH)-treated rats. One hundred eight male Sprague Dawley rats were randomly divided into 3 groups depending on the types of CLA isomers, i.e. control group (no CLA contained), c9t11 group (cis-9, trans-11 CLA contained), and t10c12 group (trans-10, cis-12 CLA contained). The experimental diet was composed of protein at 20%, carbohydrate at 56.2%, and fat at 14.5% including 1.0% CLA isomers by weight. The experimental diet was fed for 30 weeks with the initiation of intramuscular injection of DMH, which was injected twice a week for 6 weeks to give total dose of 180 mg per kg body weight. Two CLA isomers (c9, t11; t10, c12) significantly reduced tumor incidence and cell proliferation by reducing the protein expression of COX-2 and PKC, and the level of TXB2, PGE2, and DAG in colonic mucosa. However, there was no significant difference in anti-carcinogenic effect between c9t11-CLA and t10c12-CLA.

Gene Expression Profiling of 1,2-Dimethylhydrazine-Stimulated Human Umbilical Vein Endothelial Cells

The purposes of this study are to establish standard model in which endothelial cell proliferations are induced by DMH stimulation in vitro, and to analyze the gene expressions of proliferative HUVECs using DNA chip technique which could evaluate the mechanisms of angiogenesis, and the development of vascular tumors. To perform the MTT assay in 96-well microplates, 104 cells were seeded in each well which were cultured in medium. On the third day, the cells were treated with 5 different concentrations of diluted DMH from 10 to 10-3 ng/ml. Five DMH-treated groups were compared with the control group which was not treated with DMH. The optical densities in each group were measured at the time of 0, 6, 12, 24, 36, 48, and 72 hours after DMH treatment. The same experiment was repeated 9 times. Statistically significant cell proliferations were observed in 1 and 10-1ng/ml group. The RNAs were isolated from HUVECs of control group and 1ng/ml DMH-treated group, and they were used to analyze the gene expressions using DNA chip technique. One hundred and seventy-seven genes(142 of up-retulated genes and 35 down-regulated genes) were identified, and several genes were associated with VEGF and FGF production. Also DMH could affect expression of genes that involve oncogenesis. Further study should be performed to evaluate the processes of angiogenesis and morphogenesis of vascular tumors, which could be utilized in the development of new therapeutic approaches.