An in vitro antiviral evaluation of punicalagin toward influenza A virus.

Objective: Influenza complications are mild to serious, and can cause death in some cases. A great deal of attention has been paid in recent years to the development and use of new antiviral compounds to overcome drug resistance in certain strains of the influenza virus and treat the clinical implications. This study aimed to investigate the antiviral effect of punicalagin and its associated mechanism against influenza A (H1N1) virus in vitro. Materials and Methods: the ant-influenza activity of punicalagin was studied in Madin-Darby Canine Kidney (MDCK) cells using influenza virus A/Puerto Rico/8/34 (H1N1) (PR8) using Hemagglutinin assay (HA) and 50% tissue culture infective dose (TCID50). Then, the inhibition of haemagglutination, virucidal activity, inhibitory effect at different times, replication of viral RNA and expression of viral genes were investigated. Results: Punicalagin could inhibit influenza virus infection with 50% inhibitory concentration (IC50) of 3.98 µg/ml and selectivity index (SI) value of 6.1. Punicalagin decreased virus titers with an inhibitory effect on virus hemagglutination (p<0.05). Punicalagin also inhibited viral adsorption. The results of virus RNA replication and viral mRNA (NS1 and HA) expression after treatment with punicalagin showed significant suppression of viral mRNA expression but no effect on replication of viral RNA. Conclusion: The results of the present study indicated that punicalagin was effective against influenza infection most probably via inhibition of haemagglutination activity and virus binding.

The effect of celastrol in combination with 5-fluorouracil on proliferation and apoptosis of gastric cancer cell lines.

Background: Despite the availability of chemotherapy drugs such as 5-fluorouracil (5-FU), the treatment of some cancers such as gastric cancer remains challenging due to drug resistance and side effects. This study aimed to investigate the effect of celastrol in combination with the chemotherapy drug 5-FU on proliferation and induction of apoptosis in human gastric cancer cell lines (AGS and EPG85-257). Materials and Methods: In this in vitro study, AGS and EPG85-257 cells were treated with different concentrations of celastrol, 5-FU, and their combination. Cell proliferation was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The synergistic effect of 5-FU and celastrol was studied using Compusyn software. The DNA content at different phases of the cell cycle and apoptosis rate was measured using flow cytometry. Results: Co-treatment with low concentrations (10% inhibitory concentration (IC10)) of celastrol and 5-FU significantly reduced IC50 (p < 0.05) so that 48 h after treatment, IC50 was calculated at 3.77 and 6.9 µM for celastrol, 20.7 and 11.6 µM for 5-FU, and 5.03 and 4.57 µM for their combination for AGS and EPG85-257 cells, respectively. The mean percentage of apoptosis for AGS cells treated with celastrol, 5-FU, and their combination was obtained 23.9, 41.2, and 61.9, and for EPG85-257 cells 5.65, 46.9, and 55.7, respectively. In addition, the 5-FU and celastrol-5-FU combination induced cell cycle arrest in the synthesis phase. Conclusions: Although celastrol could decrease the concentration of 5-fluorouracil that sufficed to suppress gastric cancer cells, additional studies are required to arrive at conclusive evidence on the anticancer effects of celastrol.

The effect of infusion time on Echium amoenum extract -induced hepatotoxicity in vitro.

Echium amoenum is an annual herb native to the northern mountains of Iran which has medicinal application. Petals of Echium amoenum (Gole-Gavzaban) is one of the most valuable medicinal plants in Iranian folk medicine. The dry petals of E. amoenum have long been used as a sedative, tonic, anxiolytic and as a treatment for sore throat, cough and inflammation. Previous studies have shown that petals of E. amoenum contain four toxic pyrrolizidine alkaloids but conflicting results have been acquired in experimental studies investigating the hepatotoxicy of E. amoenum. However, the direct effect of E. amoenum on liver cells and the complete mechanisms of its possible cytotoxic effects toward these cells remain to be defined. The main aim of this study was to assay the mechanisms underlying the toxic effects of E. amoenum toward hepG2 cells. E. amoenum extract was obtained by infusion of dried petals in hot water (90 centigrade) for 15 or 30 min. Cell viability and mechanistic parameters were determined following 12 h incubation of hepG2 with E. amoenum extract that was obtained after 15 or 30 min infusion. The results indicated that E. amoenum extract exerts cytotoxic effects on hepG2 cells, probably through mitochondrial and lysosomal damage induced by glutathione depletion and oxidative stress.

Coevolution of HTLV-1-HBZ, Tax, and proviral load with host IRF-1 and CCNA-2 in HAM/TSP patients.

Background HTLV-1-associated myelopathy (HAM/TSP) is a progressive neurodegenerative inflammatory condition of HTLV-1 infection. Viral-host interactions are a significant contributor to the symptoms of HTLV-1-associated diseases. Therefore, in this study, the expression of the main regulatory viral factors and proviral load (PVL) and two host transcription molecules were evaluated in HAM/TSP patients. Materials and methods The study population included 17 HAM/TSP patients, 20 asymptomatic carriers (ACs), and 19 healthy controls (HCs). RNA and DNA were extracted from PBMCs for assessment of the gene expressions and PVL assessment using RT-qPCR and TaqMan method. Results HTLV-1-PVL was higher in HAM/TSPs (395.80 ± 99.69) than ACs (92.92 ± 29.41) (P = 0.001). The Tax expression in HAM/TSPs (7.8 ± 5.7) was strongly higher than ACs (0.06 ± 0.04) (P = 0.02), while HTLV-1-HBZ was only increased around three times in HAM/TSPs (3.17), compared to ACs (1.20) and not significant. The host IRF1 expression in HAM/TSPs (0.4 ± 0.31) was higher than ACs (0.09 ± 0.05) (P = 0.02) and also HCs (0.16 ± 0.07) (P = 0.5), but lower in ACs than HCs (p = 0.01). Although, in HAM/TSPs (0.13 ± 0.09) and ACs (0.03 ± 0.02) CCNA-2 expression was statistically fewer than HCs (0.18 ± 0.06) (P = 0.03, P = 0.001, respectively), in HAM/TSP was higher than ACs (P = 0.1), but did not meet a 95% confidence interval. Conclusion The study showed that HTLV-1-PVL and Tax, along with host IRF-1, could be considered biomarkers in HAM/TSP development. Furthermore, IRF-1, as an essential transcription factor, can be considered a pivotal target in HAM/TSPs treatment.

Anethole Ameliorates Acetic Acid-Induced Colitis in Mice: Anti-Inflammatory and Antioxidant Effects.

Anethole has possessed anti-inflammatory and antioxidant responses in numerous studies. Oxidative stress has a pivotal role in the pathophysiology of colitis. The current study is designed to determine the effect of anethole on acetic acid-induced colitis in mice in view of its possible anti-inflammatory and antioxidant properties. In this study, 48 mice were grouped into 6 groups (n = 8), and colitis was induced with 0.2 ml of 7% acetic acid. Mice received intraperitoneally (i.p.) for 7 constant days normal saline and/or anethole at doses of 31.25, 62.5, 125, and 250 mg/kg, respectively. After treatments, the colon was dissected out, and histopathological changes, expression of inflammatory genes (IL-1ß, TNF-α, and TLR4), and evaluation of malondialdehyde (MDA) levels and total antioxidant capacity (TAC) were assessed. The results showed that colitis is associated with edema and inflammatory responses in all layers and severe damage to the epithelium of the colon. Colitis causes a decrease in TAC, an increase in MDA levels, and an increase in inflammatory genes in the colon. Findings determined that anethole ameliorated the adverse effects of acetic acid-induced colitis in the colon. It is concluded that anethole, partially at least, possessed protective effects in acetic acid-induced colitis in mice through attenuation of oxidative stress and inflammatory response.

Pomegranate peel extract inhibits internalization and replication of the influenza virus: An in vitro study.

OBJECTIVE: Influenza virus, which is associated with high level of morbidity and mortality, has been recently considered a public health concern; however, the methods of choice to control and treat it are limited. Our previous study showed anti-influenza virus activity of pomegranate peel extract (PPE). In this study, the mechanism through which PPE acts against influenza virus A/Puerto Rico/8/34 (H1N1; PR8) was investigated. MATERIALS AND METHODS: Ethyl alcohol extract of pomegranate (Punica granatum L.) peel was prepared, and the action mechanism of PPE in inhibiting influenza replication was studied by time-of-drug-addition assay, virucidal activity, RNA replication, hemagglutination inhibition assay, viral mRNA expression, and western blot analysis. RESULTS: PPE inhibited viral polymerase activity, viral RNA replication, and viral protein expression but could not affect hemagglutination inhibition and virucidal activity. According to time-of-drug-addition assay results, PPE inhibited the virus adsorption and early steps of influenza replication. CONCLUSION: This study demonstrated that the antiviral effect of PPE on influenza virus is most probably associated with inhibition of viral adsorption and viral RNA transcription.

MicroRNA-122 in patients with hepatitis B and hepatitis B virus-associated hepatocellular carcinoma.

Hepatitis B virus (HBV) infection is known as a serious problem in the domain of public health and approximately 350 million people across the world are affected with this infectious disease. As well, microRNAs are recognized as a type of small non-coding RNAs that can be widely used as a diagnostic biomarker and prognosis method of special diseases. In this respect, microRNA-122 or miR-122 can play a significant role in the pathogenesis of several hepatic diseases. Given the importance of microRNA-122 in the liver as well as its pathology, this study focused on the potential functions of microRNA-122 in pathogenesis, diagnosis, and treatment of HBV infection. In this regard, the findings of previous studies had indicated that expression of microRNA-122 in patients with HBV infection could be significantly deregulated. The results of this study were consistent with the idea that diagnosis and treatment of this infectious disease using microRNA-122 could be an efficient method.

Anti-Influenza Virus Activity and Phenolic Content of Pomegranate (Punica granatum L.) Peel Extract and Fractions.

BACKGROUND: Influenza virus, associated with high level of morbidity and mortality, has been recently considered a public health concern while the choices for the control and treatment of the disease are limited. The present study was conducted to evaluate activity of pomegranate peel extract and its fractions against Influenza A virus in vitro . METHODS: In this research, ethyl alcohol extract of pomegranate peel was prepared and subjected to fractionation with different polarities. The potential in vitro anti-influenza A virus activity of the extract and fractions was assessed using Cytopathic Effect (CPE) reduction assay, Hemagglutinin Assay (HA), and 50% Tissue Culture Infectious Doses (TCID50) method in Madin-Darby Canine Kidney (MDCK) cells. RESULTS: The crude pomegranate peel extract and its n-butanol and ethyl acetate fractions had the highest inhibitory effect against influenza A virus with IC50 value of 6.45, 6.07 and 5.6 µg/ml in MDCK cells, respectively. Our results also showed that, the production of virus was significantly reduced upon treatment with crude extract, n-butanol and ethyl acetate fractions in a dose-dependent manner (p<0.05). CONCLUSION: Based on our results, the ethyl alcohol extract and its polar fractions of pomegranate peel can inhibit influenza A virus replication in vitro. Therefore, further characterization of its active ingredients and the mechanism of action should be carried out.

In vitro immunomodulatory activity of celastrol against influenza A virus infection.

CONTEXT: The influenza A virus (IAV) causes severe respiratory disease that remains a leading reason for morbidity and mortality. Previous studies have indicated that influenza complications in addition to viral replication are due to overproduction of pro-inflammatory cytokines. Therefore, a new compound is needed to be used with current antiviral drugs to modulate overproduction of pro-inflammatory cytokines in IAV infection. OBJECTIVE: This study investigated the effect of celastrol on mRNA expression and concentration levels of the pro-inflammatory cytokines tumor necrosis factor alpha (TNFα) and interleukin-6 (IL6) that are induced by influenza A/Puerto Rico/8/34 (H1N1; PR8) in Madin-Darby Canine Kidney (MDCK) cells. The effect of this compound on virus titration and viral mRNA expression was also investigated. METHODS: Confluent MDCK cells were infected with influenza virus (H1N1; PR8) and treated with celastrol at different concentrations. After incubation, mRNA expression and concentrations of TNFα and IL6 were investigated using real-time polymerase chain reaction (PCR) and ELISA. The viral mRNA expression and virus titration were investigated using real-time PCR and 50% tissue culture infectious dose (TCID50) assay, respectively. RESULTS: mRNA expression and concentrations of TNFα and IL6 increased significantly in control virus compared to cell control, and decreased significantly when compared with control virus after celastrol treatment. Viral mRNA expression and virus titration did not decrease after celastrol treatment. CONCLUSION: Due to reducing mRNA expression and concentrations of TNFα and IL6, celastrol can serve as a suitable choice to control cytokine-induced inflammation in IAV infection, and therefore it can be used with current antiviral drugs.

Traditional uses of medicinal plants to prevent and treat diabetes; an updated review of ethnobotanical studies in Iran.

BACKGROUND: Obesity and physical inactivity are currently on the rise due to industrialization of the communities, which has recently led to increased incidence of different diseases such as diabetes. Epidemiological studies and figures have demonstrated the growing incidence of diabetes. Relevantly, the side effects of chemical drugs have led patients to use medicinal plants and traditional approaches despite advances in development of chemical drugs. The aim of this review article is to report the medicinal plants and their traditional uses to prevent and treat diabetes according to the findings of ethnobotanical studies conducted in different regions of Iran. EVIDENCE ACQUISITIONS: The search terms including ethnobotany, ethnomedicine, ethnopharmacology, phytopharmacology, phytomedicine, Iran, and traditional medicine in combination with diabetes, blood sugar and hyperglycemic were searched from scientific databases. RESULTS: The results of this article can be a comprehensive guideline, based on ethnobotany of different regions of Iran, to prevent and treat diabetes. According to this review article, certain plant species such as Urtica dioica L., popularly called nettle, in eight regions, Teucrium polium L., popularly called poleigamander, in five regions, and Trigonella foenum-graecum L., Citrullus colocynthis (L.), Schrad., and Juglans regia L. in four regions, were reported to be frequently used to prevent and treat diabetes. CONCLUSIONS: The introduced medicinal plants in this review can be investigated in further research and produce new drugs with limited side effects.

The Radioprotective Effect of Resveratrol Against Genotoxicity Induced by γ-Irradiation in Mice Blood Lymphocytes.

In this study, we evaluated whether the protective potential of resveratrol (RSV; 3,5,4'-trihydroxy-trans-stilbene) against γ-radiation caused damages in peripheral blood lymphocyte of mice. Resveratrol as a polyphenolic compound scavenges free radicals. Various doses of RSV were administered intraperitoneally 2 hours to adult male mice before a single dose of whole-body γ-irradiation (2 Gy). To assess the protective ability of RSV, the alkaline comet assay in blood lymphocyte of mice was performed and the total comet score was evaluated. The results of the alkaline comet assay showed that RSV significantly inhibited radiation-induced DNA damage. We observed that RSV protects blood lymphocyte against radiation-induced damage in mice.

In vitro antiviral effects of Peganum harmala seed extract and its total alkaloids against Influenza virus.

This research was aimed to evaluate the in vitro antiviral effect and the mechanism of the effect of Peganum. harmala seeds extract against influenza A virus infection using Madin-Darby canine kidney (MDCK) cells. In this research, ethyl alcohol extract of P. harmala seeds and its total alkaloids was prepared. The potential antiviral activity of the extract and its total alkaloids against influenza A/Puerto Rico/8/34 (H1N1; PR8) virus was assessed. The mode of action of the extract to inhibit influenza replication was investigated using virucidal activity, hemagglutination inhibition assay, time of addition assays, RNA replication, western blot analysis and RNA polymerase blocking assay. The crud extract of P. harmala seed and its total alkaloids showed the best inhibitory effect against influenza A virus replication in MDCK cells using MTT assay, TCID50 method and hemagglutination assay. Our results indicated that the extract inhibits viral RNA replication and viral polymerase activity but did not effect on hemagglutination inhibition and virucidal activity. This study showed that, in vitro antiviral activity of P. harmala seed extract against influenza virus is most probably associated with inhibiting viral RNA transcription. Therefore, this extract and its total alkaloid should be further characterized to be developed as anti-influenza A virus agent.

In vitro and in vivo effects of Peganum harmala L. seeds extract against influenza A virus.

OBJECTIVE: Influenza A virus infections are still a major health problem and the choices available for the control and treatment of the disease are limited. This research evaluated in vitro and in vivo antiviral effects of Peganum harmala L. seeds (PHS) extract against influenza A virus. MATERIALS AND METHODS: In this research, in vitro anti-influenza A virus activity of the extract was assessed in Madin-Darby canine kidney (MDCK) cells. In order to evaluate anti-influenza activity of PHS extract in vivo, BALB/c mice were infected with 5LD50 of mouse-adapted influenza virus (H1N1; PR8) and received 200 mg/kg/day of PHS extract or 20 mg/kg/day oseltamivir. Lungs of seven mice per group were removed on day 3 post-infection and lung virus titers were determined by qRT-PCR. Mice survival, body weights and general conditions were observed for up to 14 days post-infection. RESULTS: The results demonstrated that, the ethanolic extract of PHS possesses high activity against influenza virus with IC50 value of 15.7 (CI95%:11.7-21) µg/ml in MDCK cells. Our results also showed that, oral administration of PHS extract (200 mg/kg/day) or oseltamivir (20 mg/kg/day) to infected mice, increased the survival rate, reduced body weight loss, and decreased lung virus titer. CONCLUSION: Based on our findings, P. harmala seeds extract can inhibit influenza A virus replication in vitro and in vivo. Therefore, isolation and characterization of the plant's active compounds and investigation of the underlying mechanisms of its antiviral action are highly suggested.

Anti-Herpes Simplex Virus Type-1 Activity and Phenolic Content of Crude Ethanol Extract and Four Corresponding Fractions of Quercus brantii L Acorn.

This research was aimed to evaluate anti-herpes simplex virus type-1 (anti-HSV-1) activity of crude ethanol extract and 4 corresponding fractions of Quercus brantii acorn in vitro. Crude ethanol extract was prepared and subjected to fractionation with different polarity. Anti-HSV-1 activity was evaluated on baby hamster kidney cell line using MTT assay. The inhibitory effect of the plant materials on adsorption and/or post-adsorption stages of HSV-1 replication cycle were determined. Regression analysis was used to determine 50% inhibitory concentration and 50% cytotoxicity concentration, from which selective index was calculated. Based on our results, the chloroform fraction and the crude extract had the highest effect against HSV-1 with selectivity indices of 53.8 and 48.4, respectively. The n-hexane, n-butanol, and chloroform fractions inhibited HSV-1 replication in postadsorption stage ( P < .001). The results obtained indicated that the chloroform fraction of Q brantii acorn with high inhibitory effect against HSV-1 replication could be a new promising anti-HSV-1 agent.

A review study on the effect of Iranian herbal medicines against in vitro replication of herpes simplex virus.

OBJECTIVE: There are a number of published data indicating in vitro anti-HSV activity of some of Iranian herbal extracts with no systematic review to discuss these results. Therefore, this article was aimed to review and discuss the methods carried out and the phytochemistry and bioactivity of the extracts used and also conclusions provided in these publications. MATERIALS AND METHODS: Published articles both in English (from Medline, Science Direct, EMBASE, Scopus, Pro Quest, Google scholar, Cochrane Library) and in Persian (from SID, Iran Medex and Magiran) databases, from 1966 to October 2014 were incorporated in this review. The in vitro studies that lacked CC50, IC50, were excluded. RESULTS: Only 42 published reports were found to examine Iranian herbs against HSV replication in vitro. Seventeen out of 42 studies in which 23 kinds of medicinal plants were subjected to crude extraction were included. The review of data showed that some of the herbal extracts including Hyssopus officinalis methanolic extract, Melissa officinalis aqueous extract, Quercus persica L. hydroalcoholic extract and Securigeras ecuridaca methanolic extract with selective index (SI) of 234, 877, >778 and 250, respectively were highly effective against HSV in vitro. CONCLUSION: More comprehensive studies using more advanced methods are needed to be done to achieve promising anti-HSV agents from the bioactive compounds isolated from these herbs.

Anti-adenovirus activity, antioxidant potential, and phenolic content of black tea (Camellia sinensis Kuntze) extract.

BackgroundAdenovirus (ADV) causes a number of diseases in human, and to date, no specific antiviral therapy is approved against this virus. Thus, searching for effective anti-ADV agents seems to be an urgent requirement. Many studies have shown that components derived from medicinal plants have antiviral activity. Therefore, the present study was aimed to evaluate in vitro anti-ADV activity and also antioxidant potential and total phenolic compounds of black tea (Camellia sinensis) crude extract. MethodsIn this study, the hydroalchoholic extract of black tea was prepared and its anti-ADV activity was evaluated on HEp2 cell line using MTT [3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay. The 50 % inhibitory concentration (IC50) and 50 % cytotoxicity concentration (CC50) of the extract were determined using regression analysis. Its inhibitory effect on adsorption and/or post-adsorption stages of the virus replication cycle was evaluated. To determine antioxidant activity, total phenol content, and flavonoids content of the extract, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, Folin-Ciocalteu method, and aluminum chloride colorimetric method were used, respectively. ResultsThe CC50 and the IC50 of the extract were 165.95±12.7 and 6.62±1.4 µg/mL, respectively, with the selectivity index (SI) of 25.06. This extract inhibited ADV replication in post-adsorption stage. The IC50 of DPPH radical was 8±1.41 µg/mL, compared with butylated hydroxytoluene, with IC50 of 25.41±1.89 µg/mL. The total phenol and flavonoid contents of the extract were 341.8±4.41 mg gallic acid equivalent per gram and 21.1±2.11 mg/g, respectively. ConclusionsHaving SI value of 25.06 with inhibitory effect on ADV replication, particularly during the post-adsorption period, black tea extract could be considered as a potential anti-ADV agent. The antiviral activity of this extract could be attributed to its phenolic compounds.

In vitro antiproliferative and apoptosis-inducing activities of crude ethyle alcohole extract of Quercus brantii L. acorn and subsequent fractions.

Cancer cell resistance to widely used chemotherapeutic agents is gradually developed. Natural products, mainly isolated from medicinal plants, have been considered as valuable sources for herbal anticancer drugs. The present study aimed to evaluate in vitro antiproliferative and apoptosis-inducing activities of crude ethyle alcohole extract and four fractions of Q. brantii acorn. Crude ethyle alcohole extract of Q. brantii acorn was prepared and subjected to fractionation with different polarity. Subsequently, the extract and the fractions wereevaluated for their in vitro antiproliferative activity in two cancerous (Hela and AGS) and one normal (HDFs) cell lines using MTT [3-(4, 5-dimethylthiazol-2ol) 2, 5 diphenyltetrazoliumbromide] assay. To determine whether the cytotoxicity of these compounds involved the induction of apoptosis, Hela cells were treated with IC50 concentrations of test compounds, stained with both propidium iodide (PI) and Annexin V-fluorescein isothiocyanate (FITC), and analyzed by flow cytometry. In vitro cytotoxicity assay showed that the cell viability was significantly reduced in a dose-dependent manner following treatment with crude ethyle alcohole extract and Cholophorm and n-Butanol fractions. Based on the probit regression model, antiproliferative activities of crude ethyle alcohole extract, Cholophorm fraction, and n-Butanol fraction on Hela and AGS cells and HDFs cells were significantly different (P < 0.001). The results of flow cytometric analysis showed that crude ethyle alcohole extract and two fractions of Q. brantii acorn induced early apoptotic cell death. These findings suggest that crude ethyle alcohole extract and Cholophorm and n-Butanol fractions of Q. brantii acorn suppress the proliferation of cancer cells through induction of early apoptosis.

High Prevalence of Symptomatic Hepatitis A Infection in Rural Area of Chaharmahal VA Bakhtiari Province, Iran.

INTRODUCTION: Hepatitis A virus (HAV) is the most common cause of hepatitis during childhood and is an important public health problem. Therefore, the aim of this study was to investigate an outbreak of symptomatic viral hepatitis in children and in young adults in a rural area from Chaharmahal Va Bakhtiari Province, Iran. MATERIALS AND METHODS: Serum samples from the 70 patients with icterus, who were suspected for HAV infection, referred to a therapeutic center in a central province of Iran from February to July, 2010 were tested for IgM specific antibody to HAV, using Enzyme linked Fluorscent assay (ELFA) Kit (General Biological Corp., Hsinchu, Taiwan). RESULTS: All of the 70 children had jaundice. The ELFA results showed that 48 out of 70 (68.6%) tested positive for anti-HAV specific antibody (IgM). The mean age of the individuals were 12.81+12.2 and 23 of them (32.8%) were females. There was significant relationship between seropositivity for IgM anti-HAV antibody and age group in the patients studied (p<0.05). CONCLUSION: The high number of cases identified, may indicate an outbreak of hepatitis A in this region with the children as the most susceptible age group to this symptomatic infection.

Antispasmodic effects of yarrow (Achillea millefolium L.) extract in the isolated ileum of rat.

Achillea millefolium L. is cultivated in Iran and widely used in traditional medicine for gastrointestinal disorders. The aim of this study was to determine the effect of hydroalcoholic extract of A. millefolium on the contraction and relaxation of isolated ileum in rat. In this experimental study, aerial parts of A. millefolium were extracted by maceration in ethanol 70% for 72 h. Terminal portion of ileum in 100 male Wistar rats was dissected and its contractions were recorded isotonically in an organ bath containing Tyrode solution (37 °C, pH 7.4) under one gram tension. Acetylcholine (1mM) and KCl (60mM) were used to create isotonic contractions. Propranolol and Nω-Nitro-L-arginine methylester hydrochloride (L-NAME) were used to investigate the mechanisms of action prior to giving the extract to the relevant groups. Data were compared by ANOVA and Turkey's post hoc test.. The results showed that the ileum contraction was induced by KCl and acetylcholine induced contraction was significantly reduced by A. millefolium extract. The cumulative concentrations of A. millefolium relaxed the KCl and acetylcholine induced contractions (n=14, p<0.001). The inhibitory effect of extract on contraction induced by KCl and acetylcholine was not significantly affected neither by propranolol (1µM) nor by L-NAME (100 µM). There was no significant difference in the rate of relaxation by propranolol and L-NAME between the two groups. In conclusion, A. millefolium can inhibit contraction of smooth muscle of ileum in rat, and it can be used for eliminating intestinal spasms. These results suggest that the relaxatory effect of A. millefolium on ileum contractions can be due to the blockade of voltage dependent calcium channels. In addition, the ß-adrenoceptors, cholinergic receptors and nitric oxide production are not powerful actors in inhibitory effect of A. millefolium. So, the nitric oxide and adrenergic systems may also be involved in the antispasmodic effect of A. millefolium.