Keivan Zandi

Abstract: Background: Gracilaria changii (Xia et Abbott) Abbott, Zhang et Xia, a red algae commonly found in the coastal areas of Malaysia is traditionally used for foods and for the treatment of various ailments including inflammation and gastric ailments. The aim of the study was to investigate anti-inflammatory, gastroprotective and anti-ulcerogenic activities of a mass spectrometry standardized methanolic extract of Gracilaria changii. Methods: Methanolic extract of Gracilaria changii (MeOHGCM6 extract) was prepared and standardized using mass spectrometry (MS). Anti-inflammatory activities of MeOHGCM6 extract were examined by treating U937 cells during its differentiation with 10 mu g/ml MeOHGCM6 extract. Tumour necrosis factors-alpha (TNF-alpha) response level and TNF-alpha and interleukin-6 (IL-6) gene expression were monitored and compared to that treated by 10 nM betamethasone, an anti-inflammatory drug. Gastroprotective and anti-ulcerogenic activities of MeOHGCM6 extract were examined by feeding rats with MeOHGCM6 extract ranging from 2.5 to 500 mg/kg body weight (b.w.) following induction of gastric lesions. Production of mucus and gastric juice, pH of the gastric juice and non-protein sulfhydryls (NP-SH) levels were determined and compared to that fed by 20 mg/kg b.w. omeprazole (OMP), a known anti-ulcer drug. Results: MS/MS analysis of the MeOHGCM6 extracts revealed the presence of methyl 10-hydroxyphaeophorbide a and 10-hydroxypheophytin a, known chlorophyll proteins and several unidentified molecules. Treatment with 10 mu g/ml MeOHGCM6 extract during differentiation of U937 cells significantly inhibited TNF-alpha response level and TNF-alpha and IL-6 gene expression. The inhibitory effect was comparable to that of betamethasone. No cytotoxic effects were recorded for cells treated with the 10 mu g/ml MeOHGCM6 extract. Rats fed with MeOHGCM6 extract at 500 mg/kg b.w. showed reduced absolute ethanol-induced gastric lesion sizes by > 99% (p < 0.05). This protective effect was comparable to that conferred by OMP. The pH of the gastric mucus decreased in dose-dependent manner from 5.51 to 3.82 and there was a significant increase in NP-SH concentrations. Conclusions: Results from the study, suggest that the mass spectrometry standardized methanolic extract of Gracillaria changii possesses anti-inflammatory, gastroprotective and anti-ulcerogenic properties. Further examination of the active constituent of the extract and its mechanism of action is warranted in the future.

Abstract: Japanese encephalitis (JE), a mosquito-borne viral disease, is endemic to the entire east and southeast Asia, and some other parts of the world. Currently, there is no effective therapeutic available for JE; therefore, finding the effective antiviral agent against JEV replication is crucial. In the present study, the in vitro antiviral activity of baicalein and quercetin, two purportedly antiviral bioflavonoids, was evaluated against Japanese encephalitis virus (JEV) replication in Vero cells. Anti-JEV activities of these compounds were examined on different stages of JEV replication cycle. The effects of the compounds on virus replication were determined by foci forming unit reduction assay (FFURA) and quantitative RT-PCR. Baicalein showed potent antiviral activity with IC50 = 14.28 mu g/mL when it was introduced to the Vero cells after adsorption of JEV. Quercetin exhibited weak anti-JEV effects with IC50 = 212.1 mu g/mL when the JEV infected cells were treated with the compound after virus adsorption. However, baicalein exhibited significant effect against JEV adsorption with IC50 = 7.27 mu g/mL while quercetin did not show any anti-adsorption activity. Baicalein also exhibited direct extracellular virucidal activity on JEV with IC50 = 3.44 mu g/mL. However, results of quantitative RT-PCR experiments confirmed the findings from FFURA. This study demonstrated that baicalein should be considered as an appropriate candidate for further investigations, such as the study of molecular and cellular mechanism(s) of action and in vivo evaluation for the development of an effective antiviral compound against Japanese encephalitis virus.

Abstract: Background and Objectives: Glycoproteins D (gD) and G (gG) of herpes simplex virus type 1 (HSV-1) are virus envelope glycoproteins that are able to induce HSV-1 antibody production in infected persons. Therefore, those proteins could be in interest to develop the serodiagnostic test(s) for HSV antibody detection. The aim of present study was the comparison of anti-gD and anti-gG antibodies in HSV-1 infected individuals serum samples. Materials and Methods: In this study, recombinant gD and gG were prepared and used for western blot test to detect the antibodies against HSV-1. Results: Our data showed the total gD antibody titer was higher than gG antibody titer in the HSV-1 infected patient's sera but the gG antibody titer was high significantly. Conclusions: According to our results, gD and gG can be used for designing the diagnostic laboratory tests to evaluate total antibody against HSV-1 and HSV-2.

Abstract: Background: Dengue is a serious arboviral disease currently with no effective antiviral therapy or approved vaccine available. Therefore, finding the effective compound against dengue virus (DENV) replication is very important. Among the natural compounds, bioflavonoids derived mainly from plants are of interest because of their biological and medicinal benefits. Methods: In the present study, antiviral activity of a bioflavonoid, baicalein, was evaluated against different stages of dengue virus type 2 (DENV-2) replication in Vero cells using focus forming unit reduction assay and quantitative RT-PCR. Results: Baicalein inhibited DENV-2 replication in Vero cells with IC50 = 6.46 mu g/mL and SI = 17.8 when added after adsorption to the cells. The IC50 against DENV-2 was 5.39 mu g/mL and SI = 21.3 when cells were treated 5 hours before virus infection and continuously up to 4 days post infection. Baicalein exhibited direct virucidal effect against DENV-2 with IC50 = 1.55 mu g/mL and showed anti-adsorption effect with IC50 = 7.14 mu g/mL. Conclusions: Findings presented here suggest that baicalein exerts potent antiviral activity against DENV. Baicalein possesses direct virucidal activity against DENV besides its effects against dengue virus adsorption and intracellular replication of DENV-2. Baicalein, hence, should be considered for in vivo evaluation in the development of an effective antiviral compound against DENV.

Abstract: The aim of this population-based study was to investigate the association of the percentage of body fat (BF) and high-sensitivity C-reactive protein (hs-CRP) when the infectious burden was adjusted for. A total of 1,546 subjects were randomly selected. BF was determined using bioelectrical impedance analysis. Sera were analyzed for IgG antibodies to Chlamydia pneumoniae, herpes simplex virus type 1, Helicobacter pylori, and cytomegalovirus using ELISA. Measurement of C-reactive protein (CRP) by a high-sensitivity CRP assay was performed. A linear relationship between an increase in the number of pathogens and CRP concentrations was observed (p = 0.007). Age-adjusted serum hs-CRP levels were correlated with percentage of BF in men (r = 0.28, p < 0.0001) and women (r = 0.37, p < 0.0001). In multiple regression analyses, hs-CRP showed significant correlations with percentage of BF after controlling for age, sex, cardiovascular risk factors, and the infectious burden was divided into two, three, and four pathogens [(beta = 0.24, p < 0.0001), (beta = 0.2 1, p < 0.0001), and (beta = 0.23, p = < 0.0001), respectively]. In conclusion, there was a strong association between hs-CRP and percentage of body fat independent of viral and bacterial pathogens that had been previously associated with coronary artery disease as well as carotid atherosclerosis.

Abstract: This study investigates the effects of 2-phenyl-1-benzopyran-4-one (flavone) on DENV-2 infectivity in Vero cells. Virus adsorption and attachment and intracellular virus replication were investigated using a foci forming unit assay (FFUA) and quantitative RT-PCR, respectively. Addition of flavone (100 mu g/mL) significantly increased the number of DENV-2 foci by 35.66% +/- 1.52 and 49.66% +/- 2.51 when added during and after virus adsorption to the Vero cells, respectively. The average foci size after 4 days of infection increased by 33% +/- 2.11 and 89% +/- 2.13. The DENV-2 specific RNA copy number in the flavone-treated infected cells increased by 6.41- and 23.1-fold when compared to the mock-treated infected cells. Flavone (100 mu g/mL) did not promote or inhibit Vero cell proliferation. The CC50 value of flavone against Vero cells was 446 mu g/mL. These results suggest that flavone might enhance dengue virus replication by acting antagonistically towards flavonoids known to inhibit dengue virus replication.

Abstract: Background: Epidemiological studies on genital human papilloma viruses infection (HPVs) in general population are crucial for the implementation of health policy guidelines for developing the strategies to prevent the primary and secondary cervical cancer. In different parts of Iran, there is a lack of population-based studies to determine the prevalence of HPV in the general population. The aim of this population-based study is to compare the prevalence rate of genital HPV infection among reproductive women with our previous clinic-based data, which showed a prevalence rate of 5% in women in southern Iran. Results: Using general primers for all genotypes of HPV, of 799 randomly selected women, five (0.63%, 95% CI 0.23-1.55%) tested positive for HPV DNA. Overall, seven different HPV genotypes were detected: six types (16, 18, 31, 33, 51 and 56) were carcinogenic, or "high risk genotypes" and one genotype (HPV-66) was "probably carcinogenic." Conclusions: In a population-based study, the prevalence of HPV infection among southern Iranian women was lower than that observed worldwide. However, our gynaecological clinic-based study on the prevalence of HPV infection showed results comparable with other studies in the Middle East and Persian Gulf countries. Since gynaecological clinic-based data may generally overestimate HPV prevalence, estimates of prevalence according to clinic-based data should be adjusted downward by the population-based survey estimates.

Abstract: Background xD;Dengue is a major mosquito-borne disease currently with no effective antiviral or vaccine available. Effort to find antivirals for it has focused on bioflavonoids, a plant-derived polyphenolic compounds with many potential health benefits. In the present study, antiviral activity of four types of bioflavonoid against dengue virus type -2 (DENV-2) in Vero cell was evaluated. Anti-dengue activity of these compounds was determined at different stages of DENV-2 infection and replication cycle. DENV replication was measured by Foci Forming Unit Reduction Assay (FFURA) and quantitative RT-PCR. Selectivity Index value (SI) was determined as the ratio of cytotoxic concentration 50 (CC50) to inhibitory concentration 50 (IC50) for each compound. xD; xD;Results xD;The half maximal inhibitory concentration (IC50) of quercetin against dengue virus was 35.7 μg mL-1 when it was used after virus adsorption to the cells. The IC50 decreased to 28.9 μg mL-1 when the cells were treated continuously for 5 h before virus infection and up to 4 days post-infection. The SI values for quercetin were 7.07 and 8.74 μg mL-1, respectively, the highest compared to all bioflavonoids studied. Naringin only exhibited anti-adsorption effects against DENV-2 with IC50 = 168.2 μg mL-1 and its related SI was 1.3. Daidzein showed a weak anti-dengue activity with IC50 = 142.6 μg mL-1 when the DENV-2 infected cells were treated after virus adsorption. The SI value for this compound was 1.03. Hesperetin did not exhibit any antiviral activity against DENV-2. The findings obtained from Foci Forming Unit Reduction Assay (FFURA) were corroborated by findings of the qRT-PCR assays. Quercetin and daidzein (50 μg mL-1) reduced DENV-2 RNA levels by 67% and 25%, respectively. There was no significant inhibition of DENV-2 RNA levels with naringin and hesperetin. xD; xD;Conclusion xD;Results from the study suggest that only quercetin demonstrated significant anti-DENV-2 inhibitory activities. Other bioflavonoids, including daidzein, naringin and hesperetin showed minimal to no significant inhibition of DENV-2 virus replication. These findings, together with those previously reported suggest that select group of bioflavonoids including quercetin and fisetin, exhibited significant inhibitory activities against dengue virus. This group of flavonoids, flavonol, could be investigated further to discover the common mechanisms of inhibition of dengue virus replication.

Abstract: Rotaviruses are recognized as the most common causes of severe gastroenteritis and death among children worldwide. The aim of this study was to evaluate the disease burden of rotavirus gastroenteritis and the prevalence of different G genotypes of rotaviruses circulating in children aged <5 years old who were hospitalized for acute gastroenteritis in Borazjan City, Iran. This cross sectional-descriptive study was done on 316 fecal samples collected from children aged <5 years old with acute gastroenteritis. All the stool specimens were tested for rotavirus with enzyme immunoassays (EIA). Rotavirus-positive specimens were genotyped by the Nested reverse transcription polymerase chain reaction (RT-PCR) and using different type specific primers. Out of total collected samples rotavirus infection was detected in 88 (27.85%). Of the rotavirus episodes, 79.54% occurred during the first 2 years of life, with the peak prevalence of severe rotavirus disease occurring in cold seasons. Among the common genotypes, G1 was the most predominant (52.27% of strains) and other identi�ed genotypes included non-typeable, G9 and G4, 40.90%, 4.54% and 2.27% of isolates, respectively. Because of the high frequency of rotavirus infection it is important to continue rotavirus surveillance in the other regions of Iran to determine accurately the burden of rotavirus disease and the emerging new genotypes. This will assist policy makers in decision making on rotavirus vaccine introduction.

Abstract: Organic matters in raw water have a potential to generate harmful disinfection by-products such as trihalomethanes (THMs) during the chlorination process. The objectives of this study were to investigate the trihalomethane formation potential (THMFP) in Karoon River water and to determine the effect of several factors including total organic carbon (TOC), pH, chlorine dosage, water temperature, and seasonal variation. The results showed that, among all factors, TOC and water temperature have a remarkable effect on THMFP. The experimental results from batch studies indicated that increasing of pH value yielded a greater THMFP concentration for Karoon River water. THMFP levels of Karoon River water in summer times, when water temperature exceeded 26A degrees C, were 1.2-1.6 times higher than in the spring and fall seasons, when water temperature was below 15A degrees C. It was found that the measured THMFP at Karoon River water in the spring and fall seasons were very rarely higher than 100 mu g/L.

Abstract: Background and Objectives: Streptococcus pyogenes ( S. pyogenes) is an important cause of pharyngitis. Rapid detection of this microorganism in throat specimens is essential to promptly start antibiotic therapy which could be lead to prevent complications and stop transmission of infection to other individuals. In the present study, fluorescent in situ hybridization ( FISH) was compared with culture method for the detection of S. pyogenes in throat swab specimens. Materials and Methods: One hundred eleven patients with pharyngitis were included in this study. The throat swab specimens of these patients were investigated by both conventional culturing and FISH. Results: Based on the results of this investigation, the sensitivity and specificity of FISH were 88.9% and 97.8%, respectively. Strikingly, in the specimen of one patient who had received antibiotic previous to clinical sampling, S. pyogenes was detected by means of FISH, whereas the culture method could not detect this bacterium. Conclusions: It seems that FISH is a suitable method for quick identification of S. pyogenes in throat swab specimens. When FISH is positive, culturing is not necessary. But because of the limited sensitivity of FISH for detection of S. pyogenes in throat swab specimens, culturing shoud be performed if FISH was negative.

Abstract: In vitro antiviral activities of three flavonoids; fisetin, naringenin and rutin against DENV-2 (NGC strain) were evaluated. Inhibitory effects of each compound at the different stages of DENV-2 infection were examined using foci forming unit reduction assay (FFURA) and quantitative real-time polymerase chain amplification (qRT-PCR). Fisetin, rutin and naringenin showed cytotoxic effects against Vero cells with 50% cytotoxicity (CC(50)) values of 247, > 1000, and 87 mu g/mL, respectively. Fisetin when added to Vero cells after virus adsorption inhibited DENV replication with a half maximal inhibition concentration (IC(50)) value of 55 mu g/mL and selectivity index (SI) of 4.49. The IC(50) value of fisetin was 43.12 mu g/mL with SI=5.72 when Vero cells were treated for 5 h before virus infection and continuously up to 4 days post-infection. There was no direct virucidal activity or prophylactic activity of fisetin against DENV-2. Rutin and naringenin did not inhibit DENV-2 replication in Vero cells. Naringenin however, exhibited direct virucidal activity against DENV-2 with IC(50) = 52.64 mu g/mL but the SI was <1. The present study suggests that among the flavonoids examined, only fisetin showed significant in vitro anti dengue virus replication activity.

Abstract: Background and Objectives: With due attention to the development of drug-resistant bacteria, discovering of new antibacterial compounds is needed. Algae produce numerous bioactive substances which may have pharmacological properties such as antibacterial activity. The objective of this investigation was to in vitro study of antibacterial activity of brown alga Sargassum oligocystum collected along the Bushehr coast of Persian Gulf (south west of Iran). Materials and Methods: Hot water extract, cold water extract, and hot glycerin extract were prepared. The effect of the extracts were investigated on Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 14990), Pseudomonas aeruginosa (ATCC 27853), and Escherichia coli (ATCC 25922). xD; xD;Results: Hot water extract exhibited antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa. Cold water extract and hot glycerin extract did not show antibacterial activity on any of the four test bacteria. The minimum inhibitory concentration (MIC) of hot water extract for both Staphylococcus aureus and Staphylococcus epidermidis was 3.175 mg/ml. However, the MIC of this extract for Pseudomonas aeruginosa was 9.556 mg/ml. xD; xD;Discussion: In this study gram-positive bacteria were more susceptible to hot water extract than gram-negative bacteria. Extract of Sargassum oligocystum could be a candidate for purification and further in vivo studies.

Abstract: Background: Some genotypes of human papillomaviruses can infect the genital tract and they are important infectious agents which their oncogenicity is regardable. Thus the aim of this study was to determine the prevalence of various genital human papillomaviruses (HPV) among women being subjected to routine pap smear test in Bushehr city of Iran. Results: Based on the collected data, 11(5.5%) samples were detected positive for HPV DNA and 189(94.5%) samples out of 200 samples were detected negative for HPV DNA. Meanwhile 4(2%) samples detected positive for HPV DNA by PCR were detected positive for HPV by pap smear test as well. On the other hand 5 samples which were detected positive for HPV by pap smear test didn't have HPV DNA after being tested by PCR method. Among the 11 positive samples 7 samples were identified as HPV-16, 3 samples were HPV-18 and one was HPV-53. Conclusion: Regarding the prevalence of highly carcinogen genotypes of HPV in our study determination of genital HPV prevalence among the normal population of women of Bushehr city is recommended.

Abstract: Antiviral drug resistance is one of the most common problems in medicine, and, therefore, finding new antiviral agents, especially from natural resources, seems to be necessary. This study was designed to assay the antiviral activity of curcumin and its new derivatives like gallium-curcumin and Cu-curcumin on replication of HSV-1 in cell culture. The research was performed as an in vitro study in which the antiviral activity of different concentrations of three substances including curcumin, Gallium-curcumin and Cu-curcumin were tested on HSV-1. The cytotoxicity of the tested compounds was also evaluated on the Vero cell line. The CC50 values for curcumin, gallium-curcumin and Cu-curcumin were 484.2 mu g/mL, 255.8 mu g/mL and 326.6 mu g/mL, respectively, and the respective IC50 values 33.0 mu g/mL, 13.9 mu g/mL and 23.1 mu g/mL. The calculated SI values were 14.6, 18.4 and 14.1, respectively. The results showed that curcumin and its new derivatives have remarkable antiviral effects on HSV-1 in cell culture.

Abstract: Enterococci are among prominent causes of nosocomial wound infections. Since the rapid detection of causative agents could make earlier administration of choice antibiotics and quick recovery of patients, so the application of rapid diagnostic methods is important. Therefore, this study was designed to evaluate fluorescent in situ hybridization (FISH) for the detection of Enterococcus in wound swab samples. The time needed for FISH procedure is about 3 h. Specimens taken from 33 hospitalized patients were examined by both FISH and culturing procedures. By using conventional culture, 10 of 33 wound samples were culture-positive. Out of these 10 specimens, eight were FISH-positive, but two specimens were FISH-negative for Enterococcus. The remaining 23 wound specimens were xD;Enterococcus negative according to the both methods. Therefore, the specificity of FISH was 100%; however, this method showed 80% sensitivity. Because of high specificity of FISH, the combined application of FISH and cultivation methods would be suggested for detection of enterococci from wound specimens in situations in which rapid diagnosis has an advantage in the treatment of patients.

Abstract: Background and Objectives: Antitumor drug resistance and side effects of antitumor compounds are the most common problems in medicine. Therefore, finding new antitumor agents with low side effects could be interesting. This study was designed to assay antitumor activity of the extract from brown alga Sargassum oligocystum, gathered from Persian Gulf seashore, against K562 and Daudi human cancer cell lines. Materials and Methods: The research was performed as an in vitro study. The effect of the alga extract on proliferation of cell lines were measured by two methods: MTT assay and trypan blue exclusion test. Results and Conclusion: The most effective antitumor activity has been shown at concentrations 500 mu g/ml and 400 mu g/ml of the alga extract against Daudi and K562 cell lines, respectively. The results showed that the extracts of brown alga Sargassum oligocystum have remarkable antitumor activity against K562 and Daudi cell lines. It is justified to be suggested for further research such as algal extract fractionation and purification and in vivo studies in order to formulate natural compounds with antitumor activities.

Abstract: Toxoplasmosis is a parasitic disease prevalent all over the world and it�s causal agent is Toxoplasma gondii. Toxoplasmosis is very important in pregnant women. Seroepidemiological survey of toxoplasmosis in young girls before marriage for identifying non immune girls could be used to prevent congenital toxoplasmosis. A seroepidemiological study of toxoplasma IgG-antibody in 516 high school girls was conducted. Sample cases were chosen randomly from public high school girls. Blood samples were collected and the obtained sera were examined for anti toxoplasma IgG and IgM antibodies by using ELISA method. Prevalence of toxoplasmosis in high school girls in Bushehr city was 22.1%. There was significant correlation between seropositivity of toxoplasma antibodies and factors such as, contact with cat, raw milk consumption, raw vegetables consumption and food consumption habits, but there was no significant correlation between seropositivity and availability of drinking water, keeping pet and information about toxoplasmosis. About 78% of high school girls in Bushehr city were seronegative in toxoplasma IgG antibody. It is recommended to the health managers to design the educational measures and develop the serological tests for identifying the non immune girls before marriage to prevent congenital toxoplasmosis.

Abstract: Gracilaria corticata is a red alga which can be collected from many sea coasts around the world such as China, India, Persian Gulf, etc. The Persian Gulf is a unique marine habitat infested with diverse seaweeds. The aim of the present study is to explore anticancer potential of the crude extracts from G. corticata which was collected from the Bushehr coast (South west of Iran). Here, different concentration of the aqueous extract from G. corticata was tested for probable antitumoral activity on Jurkat and molt-4 human lymphoblastic leukemic cell lines. The cells were treated by different concentration of algal extract and the number of viable cells was determined by trypan blue. Also, cytotoxicity of the extract was evaluated by methyl thiazolyl tetrazolium (MTT) assay. The results showed that 9.336 and 9.726 mu g/mu l of algal extract were the most effective concentrations against Jurkat and molt-4 cells, respectively. The water crude extract of red alga G. corticata had significant anticancer activity and it might be a good candidate for further investigations in order to develop a natural compound as an anticancer agent which can be used for the production of potential anticancer drug and novel pharmaceutical leads.

Abstract: Toxoplasmosis is a parasitic disease caused by an intracellular protozoan, Toxoplasma gondii and congenital form is one of the most important clinical aspects of this disease. Seroepidemiological studies among women of childbearing age could provide appropriate approaches to design prevention measures to prevent congenital Toxoplasmosis. In this study, 303 blood samples were collected from women of childbearing age who referred to the health center clinics for premarital examinations in Bushehr city. Sera were tested for anti-Toxoplasma IgG and IgM antibodies by using ELISA method. 71 cases (23.4%) were seropositive and 232 (76.6%) serum samples were seronegative for anti-Toxoplasma IgG antibody, 10 samples (3.3%) were seropositive and 293 samples (96.7%) were seronegative for anti-Toxoplasma IgM antibody. According to the results, there was no significant association between seropositivity and age groups, area of residency, history of contact with cat, educational levels and nutritional behaviors. Most women of childbearing age in Bushehr City do not have immunity against Toxoplasmosis. These women are at risk of acquired Toxoplasmosis in their pregnancy period. It is recommended to health managers to design the educational measures to prevent congenital Toxoplasmosis.

Abstract: Background and Objectives: Molecular epidemiology of hepatitis C virus (HCV) is very important for the treatment of hepatitis C infection. The aim of this study was to determine the distribution of HCV genotypes in Bushehr province (South West of Iran). Materials and Methods: A total of 100 patients who were detected as positive for HCV antibody (by using ELISA method and RIBA test) referred to Arya Virology Laboratory between 2007-2009 in order to molecular diagnosis and furthermore virus genotyping. After detection of HCV, RNA genotyping of virus was done by using genotype specific primers. Results: Genotype 1a was found in 49% of the patients and genotype 3a was found in 40% of the patients and 1b in 5% of patients, while the genotype of the virus could not be identified in 5% of the patients. Finally, in 1% of patients coinfection due to 1a-3a genotypes was identified. Conclusion: The dominant genotype of HCV in Bushehr province, Iran, was determined as 1a.

Abstract: Avicenniaceae family is a member of true mangrove plants which has one genus, 11 species and several subspecies. Avicennia marina is the most current species among these plants in Iranian mangrove forest. Regarding to the presence of many active biological constituents in this plant and their applications in traditional and alternative medicine, the in vitro antiviral activity of its leaf extract on herpes simplex virus type-1 (HSV-1) and vaccine strain of polio virus (Sabin) in Vero cell line were determined. The CC50 of the extract was 5750.96 for Vero cells. The antiviral effect of the extract on HSV-1 and vaccine strain of polio virus before and after the attachment of the virus particles to Vero cells were assessed. The IC50 values of the extract were 66 mu g/ml and 137.24 mu g/ml for before and after virus attachment stages of HSV-1 replication cycle respectively. The IC50 values of extract for vaccine strain of poliovirus were 145.7 and 314.3 mu g/ml for before and after attachment stages of virus replication respectively. The SI values of the extract for the before and after virus attachment stages of viral replication cycle were 87.1 and 41.9 for HSV-1. The SI for the vaccine strain of poliovirus were calculated 39.5 and 18.3 for before and post attachment stages of this virus replication cycle ordinarily. The obtained SI values indicate that hot glycerin extract of A. marina leaves could be a good candidate for further studies in the area of antiviral compound developing.

Abstract:

Abstract: Studies on curcumin, the principal element of turmeric powder, have demonstrated several biological actions such as antibacterial activity. Evaluation of new analogs or new compounds of curcumin for their antibacterial effect is interesting for researchers. In this in vitro study, we attempted to test the antibacterial activity of indium curcumin (In(CUR)(3)), indium diacetylcurcumin (In(DAC)(3)), and diacetylcurcumin (DAC) in comparison with curcumin. The action of these agents were examined on Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 14990), Pseudomonas aeruginosa (ATCC 27853), and Escerichia coli (ATCC 25922). Curcumin was effective against S. aureus and S. epidermidis, whereas In(DAC)(3) showed activity against S. epidermidis and P. aeruginosa. The effect of In(DAC)(3) on P. aeruginosa is an advantage. Strikingly, In(CUR)(3) exhibited antibacterial activity on all the four mentioned strains. DAC did not show antibacterial effect on any of the four test bacteria. The minimum inhibitory concentration (MIC) of curcumin was 187.5 mu g/ml for S. aureus, and 46.9 mu g/ml for S. epidermidis. However, the MIC of In(CUR)(3) was lower for the same bacterial strains (93.8 mu g/ml for S. aureus and 23.4 mu g/ml for S. epidermidis). Therefore, In(CUR)(3) was found to have more antibacterial effect than curcumin itself and could be a suitable candidate for further in vivo investigations.

Abstract: The hot water extract of a brown marine alga, Cystoseira myrica, from the Persian Gulf was evaluated as an antiviral compound against KOS strain of HSV-1 in cell culture. The extract exhibited antiviral activity against herpes simplex virus type 1 (HSV-1) not only during absorption of virus to the cells, but also on post attachment stages of virus replication. The water extract of C. myrica was sterilized by filtration and autoclaving, respectively. The IC50 for filtered extract was 99 mu g/ml and the IC50 for autoclaved extract was 125 mu g/ml. Based on resulted selectivity index (SI) values of the extracts, which were 33.4 and 28.2 for filtered and autoclaved extracts, respectively, we found that the antiviral compound(s) in the water extract of C. myrica to be heat stable. Also, the SI values for inhibition of the post attachment stages of HSV-1 replication were 23.1 and 21.7 for filtered and autoclaved extracts, respectively. The IC50 in this phase of study were 143 and 162 mu g/ml for filtered and autoclaved extracts, respectively. Therefore, C. myrica could be a good candidate as a natural source for anti-HSV-1 compound(s) isolation.

Abstract: In this study we tested the antiviral activity of a crude hot glycerine extract of Aloe vera gel which was grown in Bushehr (Southwest of Iran) against HSV-2 replication in Vero cell line. The extract showed antiviral activity against HSV-2 not only before attachment and entry of virus to the Vero cells but also on post attachment stages of virus replication. The IC50 before attachment and entry of virus to the cells is 428 mu g/ml and the CC50 value which is the cytotoxicity of the extract for Vero cells is 3238 mu g/ml, while the calculated selectivity index (SI) is 7.56. Also, IC50 of extract on post attachment stages of replication is 536 mu g/ml and the SI value for inhibition of the post attachment stages of HSV-2 replication is 6.04. Therefore, compounds of Aloe vera from Bushehr could be a good candidate as a natural source for antiviral drug development against HSV-2.

Abstract: Sea hares have greatly attracted the interest of all those investigating chemical defense substances. Most of these substances are low molecular weight compounds derived from algal diets. In vitro anticancer effect of a 60 kDa protein isolated from the purple fluid of Aplysia dactylomela on four human cancer cell lines was investigated in this study. A 60 kDa protein was purified from secreted purple fluid of A. dactylomela, a sea hare from Persian Gulf. The protein purification procedure consisted basically of ammonium sulfate precipitation, ion exchange chromatography using DEAE Sepharose and ultra-filtration method. In vitro antiproliferative and cytotoxic activity of the protein of interest were evaluated on L929, K562, HL60 and NB4 human cancer cell lines. The antiproliferative and cytotxic effects of 60 kDa protein on human cancer cell lines were measured by MTT assay. Results showed that the 60 kDa protein of the purple fluid of A. dactylomela exhibited the antiproliferative effect on human cancer cell lines, especially on NB4 cell line. It was maximally active at 0.5-1.5 mu g/ml on NB4 cell line. Interestingly, the protein did not show significant cytotoxic effects.

Abstract: Limited information is available concerning the role of measles-specific cell mediated immunity as a correlate of long-term protection from measles infection. Although serological responses are determined in epidemiological studies and high antibody titer is a good indicator of protection, the role of Cell-Mediated Immunity (CMI) has to be defined more clearly. In this study, Lymphocyte Proliferation (LP) and Viral Neutralization Test (VNT) were used in order to measure measles-specific cellular and humoral immune responses of 100 high school students in Tehran. From total number of subjects studied, 33 were girls and 67 were boys and all were in good health. Of these, 77 had protective neutralizing measles antibody titers and 23 did not have such titer. The results of LP showed that 89 subjects had protective cellular immune responses and 11 did not. A quantitative relationship between humoral and cellular immune responses was not observed. These findings suggest that measles-specific protective CMI is measurable for longer time in comparison to humoral immunity. These data suggest that LP responses may be better sustained than antibody titers in some children.

Abstract: The herpes simplex viruses are important causes of disease worldwide. Herpes simplex virus type 1 (HSV-1) is the primary cause of oral-facial and pharyngeal infections and may cause herpetic whitlow, eye infections as well as severe and sometimes dangerous infections of the eyes and brain. HSV-1 also accounts for 10-15% of all genital herpetic infections. Therefore, laboratory diagnosis of this virus and development of diagnostic serological techniques for HSV-1 is of particular importance. In the present study, pTrc His2A-gG1 plasmid, containing the full-length glycoprotein G (gG) protein, was produced in a prokaryotic system for the first time. Upon confirmation of a 37-kDa gG-1 protein production in a prokaryotic system based on western blotting and monoclonal antibodies, the protein was produced at a large scale and purified by ion-exchange chromatography using DEAE-sepharose. An HSV-1 type-specific diagnostic kit was designed and developed and the specificity and sensitivity of this kit were demonstrated to be 89.5% and 100%, respectively, as compared with a commercially available kit. A significant correlation was shown between the developed kit and the commercial kit.

Abstract: The metabolism and disposition of varenicline ( 7,8,9,10-tetrahydro6,10-methano-6H-pyrazino[2,3-h][3]benzazepine), a partial agonist of the nicotinic acetylcholine receptor for the treatment of tobacco addiction, was examined in rats, mice, monkeys, and humans after oral administration of [ C-14] varenicline. In the circulation of all species, the majority of drug-related material was composed of unchanged varenicline. In all four species, drug-related material was primarily excreted in the urine. A large percentage was excreted as unchanged parent drug ( 90, 84, 75, and 81% of the dose in mouse, rat, monkey, and human, respectively). Metabolites observed in excreta arose via N-carbamoyl glucuronidation and oxidation. These metabolites were also observed in the circulation, in addition to metabolites that arose via N-formylation and formation of a novel hexose conjugate. Experiments were conducted using in vitro systems to gain an understanding of the enzymes involved in the formation of the N-carbamoylglucuronide metabolite in humans. N-Carbamoyl glucuronidation was catalyzed by UGT2B7 in human liver microsomes when incubations were conducted under a CO2 atmosphere. The straightforward dispositional profile of varenicline should simplify its use in the clinic as an aid in smoking cessation.

Abstract: Herpes simplex virus produces primary and latent infections with periodic recurrency. The prime-boost immunization strategies were studied using a DNA vaccine carrying the full-length glycoprotein D-1 gene and a baculovirus-derived recombinant glycoprotein D, both expressing herpes simplex virus glycoprotein D-1 protein. Immunization with recombinant DNAs encoding antigenic proteins could induce cellular and humoral responses by providing antigen expression in vivo. Higher immune response, however, occurred when the recombinant proteins followed DNA inoculation. While all groups of the immunized mice and positive control group could resist virus challenge, a higher virus neutralizing antibody level was detected in the animals receiving recombinant protein following DNA vaccination.

Abstract: To investigate the functional relationship between the ability of the adenovirus-5 E1A oncogene product to transform with its ability to block adipocytic differentiation and induce apoptosis, we expressed E1A in the 3T3 Ll preadipocytic cell line. The results demonstrate a dramatic, quantitative reciprocal regulation of differentiation and several transformation-associated properties in response to graded levels of E1A expression, with the suppression of differentiative capacity, focus formation, and anchorage-independent proliferation requiring increasing levels of Ell A. Progressively higher E1A levels were accompanied by apoptosis induction. The effect of E1A upon adipocytic differentiation as well as transformation and apoptosis required binding to the retinoblastoma-susceptibility gene product. These data reveal a dissociation between E1A signals leading to transformation, suppression of differentiation and induction of apoptosis, based on levels of expression. (c) 2005 Wiley-Liss, Inc.

Abstract: The degradation product of ezlopitant was isolated from low specific activity material and identified by solution phase hydrogen/deuterium (H/D) exchange and electrospray ionization tandem mass spectrometry (ESI/MS/MS) to be an isopropyl peroxide analog of ezlopitant. The structure of the degradant was further confirmed by nuclear magnetic resonance (NMR) spectroscopy utilizing complete H-1 and C-13 assignments. Studies were also performed to identify the factors responsible for the oxidative degradation of ezlopitant, which included salt form, storage conditions and salt formation solvent. Of all the variable studies over a 3 weeks period, only a change in the salt form prevented this oxidative degradation. (C) 2003 Elsevier Science B.V. All rights reserved.