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Context: Today many materials have been introduced for root-end filling materials. One of them is mineral trioxide aggregate (MTA) that is mentioned as a gold standard., Aims: The purpose of this in vitro study was to evaluate the reaction of human periodontal ligament fibroblasts to the root-end filling materials, such as ProRoot MTA, Portland cement, and amalgam., Settings and Design: Eight impacted teeth were extracted in aseptic condition. The tissues around the roots were used to obtain fibroblast cells. After cell proliferation, they were cultured in the chamber slides and the extracts of the materials were added to the wells., Materials and Methods: Immunocytochemical method for measuring the expression of Fibronectin, collagen I and transforming growth factor beta (TGF®) was performed by Olysia Bioreport Imaging Software., Statistical Analysis Used: The results were analyzed by SPSS 13.0 and Tukey post hoc test with P<0.05 as the limit of significance., Results: Collagen expression in MTA specimens was higher than the other groups in 24 h significantly. After 48 h, the Portland cement group showed the most expression of collagen significantly and after 1 week, Portland cement and MTA groups had the most expression of collagen but there was no significant difference between these 2 groups. After 1 week, the Portland cement group demonstrated a higher amount of TGF® and fibronectin., Conclusions: The results suggest that Portland cement can be used as a less expensive root filling material with low toxicity. It has better effects than amalgam on the fibroblasts.

Runx3, a member of Runt-related transcription factor (Runx) proteins with tumor suppressor effect, is a tissue-restricted and cancer related transcription factor that regulate cell proliferation and growth, as well as differentiation. In the present study, exogenous Run3 was transiently expressed in AGS (human gastric adenocarcinoma), with undetectable Runx3 protein and in A549 (human lung carcinoma) with low levels of endogenous Runx3 protein. The GFP tagged Runx3 was transfected into AGS and A549 cells using fugene6 and PolyFect and Runx3 expression was confirmed by fluorescent microscopy and RT-PCR. The effect of Runx3 transfection on cell proliferation was determined by MTT assay and the results were confirmed by the trypan blue dye exclusion method. The effect of Runx3 expression on mRNA expression of BCL2-associated X protein (Bax) was evaluated using RT-PCR. In AGS and A549 cells, Runx3 expression inhibited cell proliferation (p < 0.01). The growth inhibition was less in A549 cells. We show that Runx3 expression increases Bax mRNA expression in AGS cells when compared with control (p < 0.05), but no significant differences in mRNA expression was observed in both examined cells. Runx3 expression has antiproliferative effect in AGS cell perhaps via increase in expression of Bax. The effect of Runx3 on A549 cells' viability which has endogenous level of Runx3 is not related to Bax. These findings implicate a complex regulation by Runx3 in inhibition of cell proliferation utilizing Bax.

Background and Objective: Enamel matrix proteins are involved in the development and regeneration of root cementum and in its attachment to dentin; however, the mechanisms through which this occurs have yet to be elucidated. The present study was therefore carried out to evaluate the mitogenic and proliferative responses of human periodontal fibroblast (HPLF) cells to Emdogain (EMD), and the potential role of cyclooxygenase 2 (COX-2) in this process., Material and Methods: We investigated the effects of EMD on 5-bromo-2'-deoxyuridine (BrdU) incorporation, colchicine freezing of mitosis, XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction and Trypan Blue dye exclusion, with or without celecoxibe, a selective cyclooxygenase-2 (COX-2) inhibitor; we also evaluated the expression of COX-2 mRNA and COX-2 protein in response to EMD., Results: EMD significantly enhanced mitosis in, and proliferation of, human periodontal ligament fibroblasts in a dose-dependent manner; however, there was a small increase of DNA synthesis only in response to a high dose of EMD (200 μg/mL). EMD (100 and 200 μg/mL) elicited an increase in COX-2 expression (p ≤ 0.05). Celecoxibe (20 μm) diminished the EMD-induced mitosis and proliferation of HPLF cells (p ≤ 0.05)., Conclusion: Celecoxibe hampered EMD-induced mitosis and proliferation, which, in association with EMD-increased COX-2 expression, indicates that COX-2 may be involved in the proliferative response of HPLF cells to EMD., (© 2010 John Wiley & Sons A/S.)

The aim of this study was to prepare and evaluate mucoadhesive core-shell nanoparticles based on copolymerization of thiolated chitosan coated on poly methyl methacrylate cores as a carrier for oral delivery of docetaxel. Docetaxel-loaded nanoparticles with various concentrations were prepared via a radical emulsion polymerization method using cerium ammonium nitrate as an initiator. The physicochemical properties of the obtained nanoparticles were characterized by: dynamic light-scattering analysis for their mean size, size distribution, and zeta potential; scanning electron microscopy and transmission electron microscopy for surface morphology; and differential scanning calorimetry analysis for confirmation of molecular dispersity of docetaxel in the nanoparticles. Nanoparticles were spherical with mean diameter below 200 nm, polydispersity of below 0.15, and positive zeta potential values. The entrapment efficiency of the nanoparticles was approximately 90%. In vitro release studies showed a sustained release characteristic for 10 days after a burst release at the beginning. Ex vivo studies showed a significant increase in the transportation of docetaxel from intestinal membrane of rat when formulated as nanoparticles. Cellular uptake of nanoparticles was investigated using fluoresceinamine-loaded nanoparticles. Docetaxel nanoparticles showed a high cytotoxicity effect in the Caco-2 and MCF-7 cell lines after 72 hours. It can be concluded that by combining the advantages of both thiolated polymers and colloidal particles, these nanoparticles can be proposed as a drug carrier system for mucosal delivery of hydrophobic drugs.

Epidermal growth factor (EGF) promotes proliferation in human mesenchymal stem cells (hMSCs) during in vitro propagation. In this study, we investigated the effects of PI3K/AKT, ERK1/2, P38 and JNK on EGF signalling in hMSCs. The effects of EGF on MAPKs and PI3K/AKT crosstalk were investigated by immunoblotting; cyclooxygenase-2 (COX-2) expression was studied by real-time RT-PCR; and cell proliferation was evaluated by methylthiazolyl tetrazolium bromide assay. Our results showed that EGF immediately activated all four pathways, induced proliferation and increased COX-2 expression. Interestingly, inhibition of PI3K/AKT-enhanced EGF-stimulated ERK1/2 activity, and inhibition of ERK1/2 and JNK reduced AKT phosphorylation. Furthermore, EGF-induced proliferation as well as EGF-augmented COX2 expression was hindered by ERK1/2 and p38 inhibitors. The results of this study provide evidences to be used in extended proliferation of hMSCs for cell therapy., (Copyright © 2011 John Wiley & Sons, Ltd.)

Background: The objective of this study was to develop pegylated poly lactide-co-glycolide acid (PLGA) immunonanocarriers for targeting delivery of docetaxel to human breast cancer cells., Methods: The polyethylene glycol (PEG) groups on the surface of the PLGA nanoparticles were functionalized using maleimide groups. Trastuzumab, a monoclonal antibody against human epidermal growth factor receptor 2 (HER2) antigens of cancer cells, used as the targeting moiety, was attached to the maleimide groups on the surface of pegylated PLGA nanoparticles. Nanoparticles prepared by a nanoprecipitation method were characterized for their size, size distribution, surface charge, surface morphology, drug-loading, and in vitro drug release profile., Results: The average size of the trastuzumab-decorated nanoparticles was 254 ± 16.4 nm and their zeta potential was -11.5 ± 1.4 mV. The average size of the nontargeted PLGA nanoparticles was 183 ± 22 nm and their zeta potential was -2.6 ± 0.34 mV. The cellular uptake of nanoparticles was studied using both HER2-positive (SKBR3 and BT-474) and HER2-negative (Calu-6) cell lines., Conclusion: The cytotoxicity of the immunonanocarriers against HER2-positive cell lines was significantly higher than that of nontargeted PLGA nanoparticles and free docetaxel.

Active targeting could increase the efficacy of anticancer drugs. Methotrexate-human serum albumin (MTX-HSA) conjugates, functionalized by luteinizing hormone-releasing hormone (LHRH) as targeting moieties, with the aim of specifically targeting the cancer cells, were prepared. Owing to the high expression of LHRH receptors in many cancer cells as compared to normal cells, LHRH was used as the targeting ligand in this study. LHRH was conjugated to MTX-HSA nanoparticles via a cross-linker. Three types of LHRH targeted nanoparticles with a mean particle size between 120-138 nm were prepared. The cytotoxicity of LHRH targeted and non-targeted nanoparticles were determined on the LHRH positive and negative cell lines. The internalization of the targeted and non-targeted nanoparticles in LHRH receptor positive and negative cells was investigated using flow cytometry analysis and fluorescence microscopy. The cytotoxicity of the LHRH targeted nanoparticles on the LHRH receptor positive cells were significantly more than non-targeted nanoparticles. LHRH targeted nanoparticles were also internalized by LHRH receptor positive cells significantly more than non-targeted nanoparticles. There were no significant differences between the uptake of targeted and non-targeted nanoparticles to the LHRH receptor negative cells. The active targeting procedure using LHRH targeted MTX-HSA nanoparticles could increase the anti-tumoral activity of MTX.

To evaluate the effect of chronic exposure to diazinon in the gastrointestinal tract, Caco-2 cells were made resistant by growing in low concentrations of diazinon (0.02 μM) that was gradually increased to 20 μM within 4.5 months. Resistant cells showed significant higher growth in the presence of 15, 45 and 135 μM of diazinon (96.08, 81.80 and 65.16% of control) compared to parent cells (79.71, 71.76 and 29.50% of control, respectively; p < 0.05). P-glycoprotein (P-gp) expression increased significantly in resistant cells (P-gp to beta-actin ratio 0.586 for parent and 1.255 for resistant cells, respectively; p < 0.05) without any alteration in MDR-1 mRNA level (p > 0.05).

The biological mechanisms of tooth movement are based on the response of periodontal tissues to mechanical forces. The final result of these responses is remodeling of the extracellular matrix. Tissue reactions may vary depending upon the type, magnitude and duration of the applied forces. The purpose of the present study was to analyze the effects of centrifugal force on the transcription of collagen type-I (Col-I), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of metalloproteinase- 1 (TIMP-1) genes in human periodontal ligament (PDL) fibroblasts. Human fibroblasts obtained from the PDL were cultured and subjected to centrifugal forces (36.3 g/cm(2)) for 30, 60 and 90 min continuously. This was also carried out interruptedly, three times for 30 min and six times for 15 min. The mRNAs encoding for Col-I, MMP-1, and TIMP-1 were quantified using RT-PCR. The mRNA levels of Col-I and MMP-1 were increased when continuous force was applied for 30 min and 60 min respectively. The interrupted force had almost no effect on Col-I, MMP-1 and TIMP-1 genes. These results indicate that continuous forces may have a greater effect in inducing gene expression during the remodeling process of PDL compared to interrupted forces with short rest periods.

The tumor suppressor p16(INK4a) has earned widespread attention in cancer studies since its discovery as an inhibitor of cyclin-dependent kinases (CDKs) 4/6. Structurally, it consists of four complete ankyrin repeats, believed to be involved in CDK4 interaction. According to the previous disparities concerning the importance of domains and inactivating mutations in p16, we aimed to search for the domain possessing the functional properties of the full length protein. Upon our in silico screening analyses followed by experimental assessments, we have identified the novel minimum functional domain of p16 to be the C-terminal half including ankyrin repeats III, IV and the C-terminal flanking region accompanied by loops 2 and 3. Transfection of this truncated form into HT-1080 human fibrosarcoma cells, lacking endogenous p16, revealed that it is able to inhibit cell growth and proliferation equivalent to p16(INK4a). The functional analysis showed that this fragment like p16 can interact with CDK4/6, block the entry into S phase of the cell cycle and suppress growth as indicated by colony formation assay. Identification of p16 minimum functional domain can be of benefit to the future peptidomimetic drug design as well as gene transfer for cancer therapy., (© 2010 Wiley-Liss, Inc.)

A series of substituted 2-phenylthiazole-4-carboxamide derivatives were synthesized as potential cytotoxic agents and evaluated against three human cancer cell lines including T47D (Breast cancer), Caco-2 (Colorectal cancer) and HT-29 (Colon cancer). The SAR of the arylacetamido pendent connected to the para-position of 2-phenylthiazole were explored. It was found that substitution at the 4-position by a methoxy group led to improvement of activity against Caco-2 cells while 2-methoxy substituent could maintain the high activity against HT-29 and T47D cell lines. Also, 3-fluoro analog showed good cytotoxic activity profile against all cell lines with IC(50) values less than 10 μg/mL., (Copyright © 2010 Elsevier Masson SAS. All rights reserved.)

The aim of the present work was to prepare and characterize poly(methyl methacrylate) nanoparticles coated by chitosan-glutathione conjugate so as to encapsulate insoluble anticancer drugs. Nanoparticles were synthesized through radical polymerization of methyl methacrylate initiated by cerium (IV) ammonium nitrate. Paclitaxel (PTX), a model anticancer drug, was encapsulated in nanoparticles with a maximal encapsulation efficiency of 98.27%. These nanoparticles showed sustained in vitro release of the incorporated PTX (75% of the loaded dose was released in 10 days). All nanoparticles had positive charge and were spherical, with a size range of about 130-250 nm. The PTX-loaded nanoparticles showed cytotoxicity for NIH 3T3 and T47D breast carcinoma cells, along with no cytotoxicity for two colon cell lines (HT29, Caco2)., From the Clinical Editor: The aim of this work was to prepare and characterize poly(methyl methacrylate) nanoparticles coated by chitosan-glutathione conjugate in an effort to encapsulate Paclitaxel as a model of insoluble anticancer drugs. These nanoparticles showed sustained in vitro drug release., (Copyright © 2010. Published by Elsevier Inc.)

Studies on biomedical applications of nanoparticles are growing with a rapid pace. In medicine, nanoparticles may be the solution for multi-drug-resistance which is still a major drawback in chemotherapy of cancer. In the present study, we investigated the potential cytotoxic effect of silver nanoparticles (Ag NPs) and silver ions (Ag(+)) in both parent and tamoxifen-resistant T47D cells in presence and absence of tamoxifen. Ag NPs were synthesized (< 28 nm) and MTT assay was carried out. The associated IC(50) values were found to be: 6.31 µg/ml for Ag NPs/parent cells, 37.06 µg/ml for Ag NPs/tamoxifen-resistant cells, 33.06 µg/ml for Ag(+)/parent cells and 10.10 µg/ml for Ag(+)/resistant cells. As a separate experiment, the effect of subinhibitory concentrations of Ag NPs and Ag(+) on the proliferation of tamoxifen-resistant cells was evaluated at non-toxic concentrations of tamoxifen. Our results suggested that in non-cytotoxic concentrations of silver nanomaterials and tamoxifen, the combinations of Ag(+)-tamoxifen and Ag NPs-tamoxifen are still cytotoxic. This finding may be of great potential benefit in chemotherapy of breast cancer; since much lower doses of tamoxifen may be needed to produce the same cytotoxic effect and side effects will be reduced.

This is a fullerene-based low toxic nanocationite designed for targeted delivery of the paramagnetic stable isotope of magnesium to the doxorubicin (DXR)-induced damaged heart muscle providing a prominent effect close to about 80% recovery of the tissue hypoxia symptoms in less than 24 hrs after a single injection (0.03 - 0.1 LD50). Magnesium magnetic isotope effect selectively stimulates the ATP formation in the oxygen-depleted cells due to a creatine kinase (CK) and mitochondrial respiratory chain-focusing "attack" of 25Mg2+ released by nanoparticles. These "smart nanoparticles" with membranotropic properties release the overactivating cations only in response to the intracellular acidosis. The resulting positive changes in the energy metabolism of heart cell may help to prevent local myocardial hypoxic (ischemic) disorders and, hence, to protect the heart muscle from a serious damage in a vast variety of the hypoxia-induced clinical situations including DXR side effects.

Cyclooxygenase-2 (COX-2) enzyme is believed to play a role in tumor angiogenesis, differentiation, and apoptosis. The inducible isoform of nitric oxide synthase (iNOS) also has the potential ability to damage DNA and conceivably contribute to tumor formation by a rise in nitric oxide production. Seventeen patients diagnosed with colorectal adenocarcinoma, who underwent surgical resection of the tumor, were enrolled in the study. Two macroscopic tissue samples, one from the tumor and the other from the tumor free surgical margin were collected from every patient as formalin fixed paraffin embedded blocks. Samples were analyzed for iNOS and COX-2 expression by immunohistochemistry and Western blotting. Results were digitized and semi-quantitatively analyzed. Immunohistochemistry revealed a similar pattern of expression for both iNOS and COX-2, as both were detected in tumor and epithelial cells. The mean iNOS and COX-2 levels determined by Western blotting method were significantly higher in tumor than in the tumor-free tissues (Wilcoxon signed-rank test, p < 0.001 both for iNOS and COX-2). Patients with lymph node involvement had higher levels of both enzymes in tumors (Mann-Whitney U test, p < 0.05). There was correlation between iNOS and COX-2 expression of tumor determined by immunohistochemistry and also by Western blotting (Spearman's rho test, R = 0.53, p = 0.03 and R = 0.57, p = 0.02, respectively). In conclusion, our results point out a relationship between iNOS and COX-2 expression in human colorectal adenocarcinomas and may also suggest a possible link between advanced stages of the disease and higher expression of iNOS and COX-2.

Inula oculus-christi L. (Compositae) extract was chromatographed and three sesquiterpene lactones ergolide, gaillardin and pulchellin C were isolated. The structures of these compounds were determined by analysis of their spectroscopic data, and their crystal structures were defined using X-ray crystallography; the isolation of ergolide and pulchellin C is reported for the first time from this species. These three compounds were evaluated for their in vitro cytotoxic activity against MDBK, MCF7 and WEHI164 cells; ergolide and gaillardin exhibited lower and significantly different IC50 values compared with pulchellin C (p<0.001).

The cell growth is controlled by the interaction of survival and cell growth arrest pathways as well as apoptosis mechanisms which determine the outcome of cell faith as proliferation or apoptosis. In this study, we have studied the activity of survival pathways, i.e., Akt and ERK1/2 with regard to XIAP (inhibitor of apoptosis) in serum starved and stimulated conditions. The HEK-293 cells were cultured in RPMI + 10% FBS. The cells were serum starved by switching to medium with 1% FBS for 24 h and serum stimulated by changing the medium to 10% FBS following serum starvation. The expression of p-Akt, p-ERK, Akt, ERK and XIAP was studied in various time points using western blot. The apoptosis was evaluated by DNA condensation using Hoechst 33258 and Caspase-3 assay. In serum starved condition expression of p-Akt and XIAP is very low. Serum stimulation increases p-Akt and p-ERK within 5 min and sustains a high level for 30 min. The expression of total Akt and ERK1/2 has not changed significantly for 24 h. XIAP expression starts at 6 h after serum stimulation, reaches to maximum level at 12 h and decreases to baseline within 24 h. Furthermore, serum starvation for 24 h does not induced apoptosis and DNA condensation. Taken together, the results indicate that serum activates Akt and ERK pathways earlier than XIAP expression. Furthermore, XIAP expression is low in serum starvation unlike p-ERK which suggests a survival role for ERK in serums starvation. The expression pattern of XIAP indicates induction by Akt and/or ERK activation which requires further studies.

Background and the Purpose of the Study: Non-steroidal anti-inflammatory drug-activated gene-1 (NAG-1) is involved in inflammation, apoptosis/survival and tumorigenesis as well as resistance to chemotherapy. NAG-1 protein is synthesized as pro-peptide, cleaved and secreted as mature protein. Regulation of NAG-1 is not completely discovered and increased level of NAG-1 has been reported in many cancers. The expression of NAG-1 in cancer cells could affect the progression of tumor growth. In addition the secretion of full length and mature forms of NAG-1 can influence cell proliferation in other cells. In this study the role of full length and mature forms of NAG-1 on viability of HT-1080 and MCF-7 cells were evaluated, and the cytotoxicity of celecoxib, indomethacin, tamoxifen and doxorubicin in HT1080 cells stably expressing NAG-1 were also tested., Methods: Full length and mature NAG-1 was cloned from cDNA library of HCT116 cells and stably transfected in HT1080 cells. Cells were treated with different concentrations of indomethacin, celecoxib, tamoxifen and doxorubicin and viability was assessed by MTT assay. The effect of conditioned medium of NAG-1 expressing cells on proliferation of MCF-7 and HT1080 cells were also tested., Results: The growth curves of HT1080 cells expressing full length and mature NAG-1 were not different. The viability of HT1080 cells expressing NAG-1 in the presence of indomethacin, doxorubicin and tamoxifen compared to untransfected cells was higher. The proliferation of HT1080 and MCF-7 cells were inhibited by conditioned medium of NAG-1 expressing cells in 24 and 48 hrs., Major Conclusion: NAG-1 expression enhances drug resistance to tamoxifen, indomethacin and doxorubicin in HT1080. In addition, condition medium of NAG-1 expression cells inhibits proliferation in MCF-7 and HT1080 cells. Thus, NAG-1 expression can induce drug resistance in NAG-1 expressing cells and inhibition of viability in non expressing cells. Thus, NAG-1 is suggested as a marker for effective cancer chemotherapy and tumor progression.

The aim of this study was to investigate the in vitro cytotoxic activity of total extract of MeOH (70%) and partition fractions of hexan, chloroform (CHCL3), ethylacetate (EtOAc) and MeOH-H2O of brown algae species (Sargassum swartzii, Cystoseira myrica, Colpomenia sinuosa) found in the Persian Gulf against in different cell lines including HT-29, Caco-2, T47D, MDA-MB468 and NIH 3T3 cell lines by MTT and AnnexinV-PI assay. The hexan fraction of S. swartzii and C. myrica showed selective cytotoxicity against proliferation of Caco-2 cells (IC50 < 100 μg/ml) T47D cell line (IC50<100 μg/ml), respectively. S. swartzii and C. myrica were also observed for increasing apoptosis in Caco-2 and T47D cells. Total extract and fractions of C. sinuosa did not show any significant cytotoxicity against the studied cell lines. MDA-MB468 cells were more sensitive to C. myrica than was T47D (IC50 99.9 ± 8.11 vs. 56.50' ± 0.88). This reflects an estrogen receptor independent mechanism for cytotoxicity of the extract. The IC50 of the hexan fraction of C. myrica on T47D parent cells was lower than it was on T47D-TR cells (IC50 99.9 ± 8.11 vs. 143.15 ± 7.80). This finding suggests a role for the MDR-1 in the development of possible future tolerance to the extract.

The present study was undertaken to explore the possible biochemical activities of Hyaenanche globosa Lamb. and its compounds. Two different extracts (ethanol and dichloromethane) of four different parts (leaves, root, stem, and fruits) of H. globosa were evaluated for their possible antibacterial, antityrosinase, and anticancer (cytotoxicity) properties. Two pure compounds were isolated using column chromatographic techniques. Active extracts and pure compounds were investigated for their antioxidant effect on cultured 'Hela cells'. Antioxidant/oxidative properties of the ethanolic extract of the fruits of H. globosa and purified compounds were investigated using reactive oxygen species (ROS), ferric-reducing antioxidant power (FRAP), and lipid peroxidation thiobarbituric acid reactive substance (TBARS) assays. The ethanolic extract of the leaves and fruits of H. globosa showed the best activity, exhibiting a minimum inhibitory concentration (MIC) of 3.1 mg/ ml and a minimum bactericidal concentration (MBC) of 1.56 and 6.2 mg/ml, respectively, against M. smegmatis. The ethanolic extract of the fruits of H. globosa (F.E) showed the highest percentage of inhibitory activity of monophenolase (90.4% at 200 mug/ml). In addition, F.E exhibited 50% inhibitory concentration (IC(50)) of 37.7 mug/ml on the viability of 'HeLa cells' using cytotoxicity MTT assay. Subsequently, F.E was fractionated using phase-partitioning with n-hexane, ethyl acetate, and n-butanol. The cytotoxicity of these fractions were determined in vitro using different cancer cell lines. The n-hexane fraction exhibited the highest activity of toxicity. Therefore, this fraction was subjected to further separation by chromatographic methods. Two pure compounds known as: 'Tutin' and 'hyenanchin' were isolated and their structures were determined by NMR spectroscopic methods. Unpredictably, none of them showed significant (P < 0.01) inhibition on cell viability/proliferation at the concentrations that were used. F.E showed significant anti-tyrosinase, antibacterial, and cytotoxicity effects, therefore it can be considered as an effective inhibitor alone or in combination with other plant extracts.

Background and the Purpose of the Study: Etoposide is an antineoplastic agent used in multiple cancers. It is known that etoposide induce cell death via interaction with topoisomerase II; however, the etopoisde cellular response is poorly understood. Upon etoposide induced DNA damage, many stress signaling pathways including JNK are activated. In response to DNA damage, it has been shown that WWOX, a recently introduced tumor suppressor, can be activated. In this study the activation of WWOX and JNK and their interaction following etoposide treatment were evaluated., Materials and Methods: HEK293 cells treated with etoposide were lysed in a time course manner. The whole cell lysates were used to evaluate JNK and WWOX activation pattern using Phospho specific antibodies on western blots. The viability of cells treated with etoposide, JNK specific inhibitor and their combination was examined using MTT assay., Results: Findings of this study indicate that WWOX and JNK are activated in a simultaneous way in response to DNA damage. Moreover, JNK inhibition enhances etoposide induced cytotoxicity in HEK293., Conclusion: Taken together, our results indicate that etoposide induces cytotoxicity and WWOX phosphorylation and the cytotoxicty is augmented by blocking JNK pathway.

Docetaxel (DTX) is one of the most effective antineoplastic drugs. However, its current clinical administration, formulated in tween80, causes serious side effects. This study is focused on preparation and evaluation of poly(lactide-co-glycolide) nanoparticles (NPs) containing DTX to remove tween80. Drug encapsulation efficiency, in-vitro drug release, cellular cytotoxicity, and in-vivo biodistribution of NPs in mice after intravenous administration were investigated. The average diameter of the NPs was approximately 172-178 nm with encapsulation efficiency of 68%. A burst release of approximately 30% (w/w) of the loaded drug followed by a sustained release profile was observed. Cellular mortality of the NPs was more than or at least as great as DTX free drug; for example, cell viability measured at 100 nmol/l drug concentration was decreased from 50.9% for DTX free drug to 15.9% for the NP formulation after 48 h incubation with T47D cells. The DTX plasma amount remained at a good level (13% of the initial dose) in the NP formulation compared with the DTX conventional formulation, which is approximately 0.5% of the initial dose, was present in plasma up to 2 h. Poly(lactide-co-glycolide) NPs containing DTX prepared in this study may be regarded as a suitable and superior formulation for the current formulation in the market containing tween80 with improved cancerous cell mortality and biodistribution characteristics.

Vinca minor L. (Apocynaceae) is a medicinal plant that has long been used to treat cerebral and memory disorders in European folk medicine. Furthermore, it contains more than 50 alkaloids, some of them having bisindole structure such as the antineoplastic alkaloids present in Catharanthus roseus (L.) G. Don (Apocynaceae). In this study, the plant's alkaloid extract was divided into three fractions and the cytotoxic effects on cell proliferation of HT-29, Caco-2, T47D, and NIH/3T3 cell lines were examined. All alkaloid fractions showed a dose-dependent cytotoxic effect on the cell lines. IC(50) values confirmed that the growth and proliferation of NIH/3T3 cells were less affected in comparison to other cell lines.

Introduction: Although islet isolation and transplantation techniques have improved extensively in recent years, the loss of healthy functional islets is one of the major obstacles in this enterprise. A biostimulatory effect of low-level laser irradiation has been proven on proliferation of some kinds of cells. The aim of this study was to evaluate the effect of low-level laser irradiation on the function of isolated rat pancreatic islets after 24 hours of preculture., Methods: Pancreatic islets isolated from male rats (250 to 300 g) were cultured for 24 hours in RPMI 1640 media. Groups of islets then received different energy densities (1, 3, 5 joules/cm(2) or silent) at 2 wavelengths (810 nm and 630 nm) using laser devices. Insulin concentrations in buffer media were measured as indices of islet function., Results: Irradiation of incubated islets with 830 nm low-level laser significantly increased insulin secretion after a glucose challenge test (P < .05). There was a significant increase in insulin secretion after irradiation with joules/cm(2) 630 nm energy density (P < .001)., Conclusion: These findings suggest that low-level laser irradiations improved islet cell function before transplantation.

Docetaxel (DTX) is one of the most active chemotherapeutic agents for treating metastatic breast cancer. Its aqueous solubility is very low, hence the available formulation of DTX for clinical use consists of high concentrations of tween80, which has been associated with several hypersensitivity reactions. To reduce the systemic toxicity of DTX as well as to avoid the use of tween80, in this study DTX was chemically conjugated with human serum albumin via a succinic spacer. A high-performance liquid chromatography method was developed for the determination of DTX-albumin conjugate. T47D and SKOV3 cells were used for the evaluation of the in vitro cytotoxicity of the conjugate by MTT assay. Studies were then done on balb/c mice to elucidate the tissue distribution of conjugates after intravenous administration. The albumin-conjugated formulation of DTX with the particle size of 90-110 nm showed enhanced solubility and in vivo characteristics and significantly higher cytotoxicity against tumor cells, for example, IC50 of 6.30 +/- 0.73 nM for T47D cell line compared to free DTX with IC50 of 39.4 +/- 1.75 nM. Conjugation also maintained DTX plasma level at 16.19% up to 2 h after injection compared with 2.51% for Taxotere, hence increasing the chance of nanoparticles uptake by tumor cells., (Copyright 2009 Wiley-Liss, Inc.)

Hyperglycaemia has been observed with exposure to organophosphate insecticides. This study was designed to compare the effects of calcium channel blockers, alpha-adrenergic, beta-adrenergic, and muscarinic receptor blockers, and of free radical scavengers on insulin secretion from diazinon-treated islets of Langerhans isolated from the pancreas of rats using standard collagenase digestion, separation by centrifugation, and hand-picking technique. The islets were then cultured in an incubator at 37 degrees C and 5 % CO2. In each experimental set 1 mL of 8 mmol L(-1) glucose plus 125 microg mL(-1) or 625 microg mL(-1) of diazinon were added, except for the control group, which received 8 mmol L(-1) glucose alone. The cultures were then treated with one of the following: 30 micromol L(-1) atropine, 100 micromol L(-1) ACh + 10 micromol L(-1) neostigmine, 0.1 micromol L(-1) propranolol, 2 micromol L(-1) nifedipine, 50 micromol L(-1) phenoxybenzamine, or 10 micromol L(-1) alphatocopherol. In all experiments, diazinon significantly reduced glucose-stimulated insulin secretion at both doses, showing no dose dependency, as the average inhibition for the lower dose was 62.20 % and for the higher dose 64.38 %. Acetylcholine and alpha-tocopherol restored, whereas atropine potentiated diazinon-induced hyposecretion of insulin. Alpha-, beta- and calcium channel blockers did not change diazinon-induced effects. These findings suggest that diazinon affects insulin secretion mainly by disturbing the balance between free radicals and antioxidants in the islets of Langerhans and by inducing toxic stress.

Aim: To assess the hepatic changes after induction of different periods of renal ischemia., Methods: Rats were subjected to either sham operation or ischemia (30, 45 and 60 min) followed by 60 min reperfusion. Liver and renal functional indices were measured. Hepatic glutathione (GSH) and ferric reducing antioxidant power levels and the concentration of interleukin (IL)-10 and tumor necrosis factor (TNF-alpha) were evaluated. Portions of liver and kidney tissues were fixed for histological evaluation., Results: Forty-five minutes renal ischemia followed by 60 min reperfusion caused significant changes in liver structure and a significant reduction in renal function. These rats showed a significant decrease in liver GSH, as well as a significant increase in TNF-alpha and IL-10 concentrations. These results demonstrated that renal ischemia caused changes in liver histology, function, oxidative stress and inflammatory status, which led to a reduction in hepatic antioxidant capacity. With 30 min ischemia, the magnitude of these changes was less than those with 45 or 60 min ischemia., Conclusion: A minimum of 45 min ischemia is needed to study the effects of renal injury on the liver as a remote organ.

A quantitative approach has been proposed to evaluate the competitive inhibition of Escherichia coli and Salmonella typhi by live and heat-inactivated laboratory isolated Lactobacillus sp. on adhesion to monolayer of Caco-2 cells. Three species of Lactobacillus (L. casei, L. acidophilus, L. agilis) isolated from human neonate feces and two commercial probiotic strains (L. casei, L. acidophilus) have been compared for probiotic activity. All lactobacilli were able to attach to the Caco-2 cells, however, the degree of adhesion was bacterial strain-dependent. The adhesion indices of the two commercial probiotic strains were not significantly different from the values obtained for the other two similar fecal strains (p > 0.01). The inhibition of attachment of the pathogenic bacteria by inactivated cells of fecal L. acidophilus was examined and compared to the results of live bacteria. The inhibition pattern was similar for live and heat-inactivated L. acidophilus (p > 0.01). The number of attached pathogenic bacteria to the Caco-2 cells decreased when the number of L. acidophilus increased from 10(6) to 10(9) CFU/mL. The heat-inactivated L. acidophilus displayed similar probiotic activity compared to the live bacteria.

Several new acenaphtho[1,2-b]quinoxaline derivatives were prepared by the reaction of o-phenylenediamines with acenaphthenequinones. The response of different carcinoid cell lines to these compounds were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide) assay and trypan blue exclusion tests. The cytotoxicity of 3,4-dinitroacenaphtho[1,2-b]quinoxaline (IIId) on the tested cell lines was confirmed by both tests. Furthermore, the MTT test showed a significant difference (p < 0.05) between the cytotoxicity of this compound on malignant cell lines of Caco-2, HT-29, T47D and non malignant mouse fibroblast cell line of NIH-3T3. An apoptosis inducing effect of compound IIId on K562 cells was detected by flow cytometry using Annexin-V-fluorescein isothiocyanate (AnV-FITC) and propidium iodide (PI) staining. The apoptosis induction (PI-/AnV+) in treated K562 cells was significantly (p < 0.01) more at 0.5 microg/ml concentration of compound IIId in comparison to all other concentrations of this compound and also doxorubicin (CAS 25316-40-9) (250 nM).

Tamoxifen causes a mitochondrial transmembrane potential dysfunction and ATP depletion, which may play a role in tamoxifen cytotoxicity. Administration of oligomycin-2 deoxy glucose (2DG) enhanced tamoxifen antiproliferative effects, which may be due to exacerbated ATP depletion following tamoxifen and oligomycin-2DG coadministration. Sodium nitroprusside (SNP) did not significantly change tamoxifen responsiveness at 0.1, 0.5, and 1 mM; however, 2 mM SNP hampered tamoxifen effects on cell proliferation and cell cycle. Oligomycin-2DG neither changed iNOS expression nor altered its attenuated expression due to tamoxifen exposure, suggesting that ATP depletion-mediated sensitivity to tamoxifen seems to be apart from iNOS.

Protein-protein interactions studies can greatly increase the amount of structural and functional information pertaining to biologically active molecules and processes. The information obtained from such studies can lead to design and application of new modification in order to obtain a desired bioactivity. Many application packages and servers performing docking, such as HEX, DOT, AUTODOCK, and ZDOCK are now available for predicting the lowest free energy state of a protein complex. In this study, we have focused on cyclin-dependent kinase 4 (Cdk4), a key molecule in the regulation of cell cycle progression at the G(1)-S phase restriction point and p16(INK4a), a tumor suppressor which inhibits Cdk4 activity. Truncated structures were created to find the more critical regions of p16 for interaction. The tertiary structures were determined by ProSAL, GENO3D Web Server. We evaluated their interactions with Cdk4 using two docking systems, HEX 4.5 and DOT 1. Calculations were performed on a high-speed computer. Minimizations and visualizations were carried out by PdbViewer 3.7. Considering shape and shape/electrostatic total energy, structures containing ANK II, III and IV motifs that lack the N-terminal region of the full length p16 molecule showed the best fit complexes among the p16 truncated forms. The free energies were compatible with that of p16 full length original form, the full length. It seems that the N-terminal of the molecule is not crucial for the interaction since the truncated structure containing only this region did not show a good total energy.

Recombinant human Factor IX (rFIX) was cloned in a mammalian expression vector and transfected into CHO and HEK-293. Treatment with 10(-9) M methyl testosterone increased rFIX production by 30-50% in CHO and HEK clones. However, 10(-9) M 17beta-oestradiol increased production of rFIX by ~50% in CHO-F7 clone and decreased production by 48% and 37% in CHO-F8 and HEK-F2-6, respectively. Progesterone treatment inhibited rFIX production in both cell lines. Production of rFIX can thus be increased by sex hormone treatment and therefore used to enhance biotechnological production in mammalian cells.

For folate-receptor-targeted anticancer therapy, docetaxel (DTX) nanoparticles (NPs) were produced employing polylactide-co-glycolide-polyethylene glycol-folate (PLGA-PEG-FOL) conjugate. The FOL-conjugated di-block copolymer was synthesized by coupling the PLGA-PEG-NH(2) di-block copolymer with an activated folic acid. It was expected that FOL moieties were exposed on the micellar surface. The conjugates assisted in the formation of DTX NPs with an average size of 200 nm in diameter through an emulsification/solvent diffusion method. The FOL-targeted NPs showed a greater extent of intracellular uptake in FOL-receptor-positive cancer cells (SKOV3) in comparison with the non-targeted NPs, indicating that the FOL-receptor-mediated endocytosis mechanism could have a role in the cellular uptake of NPs. These results suggested that FOL-targeted DTX NPs could be a potentially useful delivery system for FOL-receptor-positive cancer cells.

Damage to the leech or mammalian CNS increases nitric oxide (NO) production and causes accumulation of phagocytic microglial cells at the injury site. Opioids have been postulated to modulate various parameters of the immune response. Morphine and leech morphine-like substance are shown to release NO and suppress microglial activation. Regarding the known immuno-modulatory effects of selective mu and kappa ligands, we have assessed the effect of these agents on accumulation of microglia at the site of injury in leech CNS. Leech nerve cords were dissected, crushed with fine forceps and maintained in different concentrations of opiates in culture medium for 3 h and then fixed and double stained with Hoechst 33258 and monoclonal antibody to endothelial nitric oxide synthase (NOS). Morphine and naloxone (> or =10(-3) M) but not selective mu agonist, DAMGO [d-Ala2, N-Me-Phe-Gly5(ol)-enkephalin] and antagonist, CTAP [D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2] inhibited the microglial accumulation. The effect of morphine was abrogated by pre-treatment with naloxone and also non-selective NOS inhibitor, l-NAME [N(omega)-nitro-l-arginine-methyl-ester; 10(-3) M] implying an NO-dependent and mu-mediated mechanism. These results are similar to properties of recently found mu-3 receptor in leech, which is sensitive to alkaloids but not peptides. Both selective kappa agonist, U50,488 [3,4-dichloro-N-methyl-N-(2-(1-pyrrolidinyl)cyclohexyl)-benzeneacetamide; > or =10(-3) M], and antagonist, nor-binaltorphimine (nor-BNI; > or =10(-3) M), inhibited the accumulation. The effect of nor-BNI was reversed by l-NAME. Immunohistochemistry showed decreased endothelial NOS expression in naloxone and U50,488-treated cords. Since, NO production at the injury site is hypothesized to act as a stop signal for microglias, opioid agents may exert their effect via changing of NO gradient along the cord resulting in disruption of accumulation. These results suggest an immuno-modulatory role for mu and kappa opioid receptors on injury-induced microglial accumulation which may be mediated via NO.

Two novel dual functional agents, 3[3-(2,4-dinitro-phenylamino)-propyl]-5-fluoro-1H-pyrimidine-2,4-dione 7 and N-[3-(2,4-dinitro-phenylamino)-propoxy]urea 8, resulting from linkage of 2,4-dinitrophenylamine through three carbon atoms with 5-fluorouracil 5 and hydroxyurea 6, respectively, were prepared and their in vitro aerobic cytotoxicities in HT-29 cell line with and without radiation were determined. Compounds 7 and 8 unlike their components were not cytotoxic but showed radiosensitizing activity.

Previous studies showed that malathion induces hyperglycemia mainly through influence on glucose metabolism in liver and skeletal muscles. The main objective of the present study was to determine what will happen on pancreatic key enzymes of insulin secretion, including glucokinase (GK) and glutamate dehydrogenase (GDH), if animals would be in acute or subchronic exposure to various doses of malathion, an organophosphorous insecticide in rats. In the subchronic study, malathion was administered orally at doses of 100 to 400 ppm for 4 weeks. In the acute experiment, animals received various doses of 3 to 75 mg/kg of malathion intraperitoneally. In each experiment, islets were isolated from the pancreas of rats by a standard collagenase digestion, separation by centrifugation, and hand-picking technique. The activities of the mitochondrial GDH and the nonmitochondrial GK enzymes were determined in islets homogenates spectrophotometrically. Blood sample was taken by cardiac puncture for glucose and insulin assays. In the acute experiment, malathion (3, 15, 75 mg/kg) increased blood glucose and insulin (15 and 75 mg/kg). In the subchronic experiment, malathion (100, 200, 400 ppm) increased blood glucose and insulin (200 and 400 ppm). All doses in both acute and subchronic experiments increased the mitochondrial GDH activity. Acute (15 and 75 ppm) and subchronic (200 and 400 ppm) increased the islets GK activity. It was concluded that pancreatic islet key enzymes are stimulated following acute and subchronic exposure to malathion though not enough to overcome to hyperglycemia. Activation of islets muscarinic receptors by malathion in favor of hyperinsulinemia, overproduction of glutamate/glutathione by GDH, and overproduction of glucose via increased glycogenolysis in counteracting with malathion-induced oxidative stress are possible mechanisms for observed effects. A new NOAEL acceptable daily intake must be established for malathion.

In this study, we investigated the effects of intrahippocampal infusion of indomethacin as a non-selective cyclooxygenase inhibitor and celecoxib as a selective cyclooxygenase-2 inhibitor on spatial memory in the Morris water maze. Rats were trained for 3 days; each day included two blocks, and each block contained 4 trials. Tests were performed 48 h after surgery. Bilateral intrahippocampal infusion of indomethacin (0.01, 0.1, or 1 M) did not show any significant effect on spatial memory retention at these concentrations in rats. We also examined effects of infusion of celecoxib (0.02, 0.06, or 0.1 M) on memory retention. Bilateral infusion of 0.1 M celecoxib significantly altered escape latency and traveled distance in rats. These results strongly suggest that cyclooxygenase-2 is involved in spatial memory retention.

Regarding the widespread use of organophosphorous pesticides (OP) especially malathion in environment and reported cases of muscle disturbances in human and animal, the present work was undertaken to explore effects of malathion subchronic exposure on rat leg skeletal muscle glucose metabolism by measuring key enzymes of glycogenolysis and glycolysis. Malathion was administered through food for 4 weeks at concentrations of 100, 200, and 400ppm to rats. Activities of enzymes including glycogen phosphorylase (GP), hexokinase (HK), and phosphofructokinase-1 (PFK) were measured in skeletal muscle homogenate of exposed rats. Levels of glucose and insulin were measured in blood. Four weeks administration of malathion at doses of 200 and 400ppm increased blood glucose concentrations to 44.4 and 60.6% of control, respectively. Malathion at doses of 200 and 400ppm increased blood insulin concentration to 36.6 and 143.2% of control, respectively. Malathion at doses of 100, 200, and 400ppm increased muscle PFK activity to 40.4, 53.5, and 83.1% of control, respectively. Malathion at doses of 400ppm increased skeletal muscle GP to 91.6% of control. Skeletal muscle HK was not influenced by malathion treatment. It is concluded that malathion influences muscle glycogenolysis and glycolysis as well as secretion of insulin from pancreas which all may explain diabetic potential of malathion.

The use of FEO as a remedy for control of primary dysmenorrhea increases concern about its potential teratogenicity due to its estrogen-like activity. Limb bud mesenchymal cells, when grown in high-density cultures, can be differentiated into a number of cell types including cartilage and muscle. These cells have been used extensively for in vitro studies of chondrogenesis. Therefore, we used limb bud cells and Alcian blue staining method that is specific for staining cartilage proteoglycan, to determine the teratogenic effect of FEO. Limb bud cells obtained from day 13 rat embryo were cultivated and exposed to various concentrations of FEO for 5 days at 37 degrees C and the number of differentiated foci were counted. Retinoic acid (90 microg/ml) was chosen as positive standard control. The differentiation was also evaluated using limb bud micromass culture using immunocytochemical techniques and BMP-4 antibody. The results showed that FEO at concentration as low as 0.93 mg/ml produced a significant reduction in the number of stained differentiated foci. However, this reduction was due to cell loss, determined by neutral red cell viability assay, rather than to be related to decrease in cell differentiation. These findings suggest that the FEO at the studied concentrations may have toxic effect on fetal cells, but there was no evidence of teratogenicity.

Insecticides are important candidates for immunotoxicology tests in order to assess the 'No Observable Adverse Effect Level' (NOAEL). Propoxure (PPX), as a carbamate household insecticide, has been used for several decades around the world in agriculture. However, there has been no clear investigation on its immunotoxic potentials. In this study, as Tier I of immunotoxicological screening tests, we examined the effects of subacute exposure to intraperitoneally (i.p.) administered PPX, at doses of 10, 2, and 0.2 mg/kg, on C57Bl/6 female mice. After 28 days administration of PPX, the treated animals were sacrificed and peripheral blood samples were collected. Then spleen (SP), thymus (TM), and bone marrow were collected and weighed. Functional tests including SRBC-hemagglutination (HA), plaque-forming colony assay (PFC), and delayed type hypersensitivity (DTH) response to SRBCs were performed. Furthermore, spleen T-cell phenotype (CD4/CD8) was also determined. Results showed that high doses of PPX could not only produce histopathological changes in TM and SP but also suppress humoral response to antigen (SRBCs). PPX at medium doses (2 mg/kg) did not show histopathological changes in TM or SP but increased the humoral response to SRBCs, as identified by HA and PFC tests. PPX at low doses (0.2 mg/kg) did not produce any significant changes in humoral (PFC and HA) or cellular responses (DTH) of the immune system. It seems that PPX has no adverse effects on mice immune system at low doses of 0.2 mg/kg/day, which is 10 times the PPX allowed daily intake (ADI) limit.

Diazinon (DZN), an organophosphate insecticide, has been used in agriculture for several years. It is possible the residue of this compound to be recycled in the biological system. There is no report on DZN immunotoxicity potential. In the present study, we examined the immunotoxic effects of intraperitoneally administered DZN in the C57bl/6 female mice. Diazinon was administered at doses of 25, 2, and 0.2 mg/kg for 28 days (five injections per week). Animals were then sacrificed to observe the cellularity or histopathological changes in thymus, spleen, bone marrow, and peripheral blood. Furthermore, humoral and cellular functional responses such as Hemagglutination titration (HA), IgM-Plaque Forming Colony Assay (PFC), Delayed-Type-Hypersensitivity (DTH) to SRBC, and T cell subtyping (CD4/CD8) were determined. The results showed that DZN at 25 mg/kg not only could produce gross histopathological changes in thymus and spleen but also could suppress both humoral and cellular activity of the immune system. At lower doses (0.2 and 2 mg/kg) there were no observable alteration in cellularity or histology of immune tissues. However, DZN at medium dose (2 mg/kg per day) could inhibit RBC-cholinesterase and showed a mild decrease (P < 0.1) in thymus/body-weight ratio and DTH response. At dose of 0.2 mg/kg no histopathological or functional disturbances were detectable. These results indicate that DZN has immunosuppressive effects in the C57bl/6 mice at doses more than 2 mg/kg. The present results however indicate that under recommended Allowed Daily Intake (ADI) limit (<0.02 mg/kg), no observable immunotoxicity effect is expected.

Studies suggest that some lead-induced toxic effects may occur through free radical production and oxidative stress. This study examined the relationship between brain histopathological alterations and oxidative stress in subchronic lead exposure. Male Albino rats received lead acetate at 0.01%, 0.05% and 0.1% w/v in their drinking water for 30 days. Animals given sodium acetate (0.1% w/v) served as control in the same period. At the end of exposure, blood-lead levels, blood catalase (CAT) and superoxide dismutase (SOD) activities and malondialdehyde (MDA) content (in blood and brain) were measured. The brain tissue samples were prepared and analysed by light and scanning electron microscopy. The results show that, the blood-lead levels in treated animals were higher in comparison with control. CAT and SOD activities in animals treated with 0.01% and 0.05% w/v did not increase in comparison with control (P > 0.05) but these values were higher in animals treated with 0.1% w/v lead acetate (P < 0.01). MDA content in blood and brain of animals treated with lead acetate 0.1% w/v, increased significantly (P <0.01), but these values were not significantly increased in other treated animals. No major histopathological alterations were detected in the brains of animals treated with lead acetate at 0.01% and 0.05% w/v. In animals treated with lead acetate 0.1% w/v, demyelinization and collagenous scar formation with neuronal atrophy in hippocampus region was observed. It is concluded that lead acetate induce oxidative stress which has an important role in brain damage in rats.

In cases of tooth avulsion, immediate replantation is the treatment of choice. In the event of any problem, the tooth should be stored in a medium that supports the periodontal ligament cell viability. Theoretically, mitotic promoting agents may be of use to preserve cell viability. It has previously been reported that levodopa (L-dopa) accelerates healing by increasing the growth hormone level. In this study, the direct effect of L-dopa was investigated on human periodontal ligament fibroblast (HPLF) cells. Cell culture methods were used to assess the potential mitogenic effect of L-dopa. Results show that in low concentrations of L-dopa (1 microgram/microl in this study), the number of HPLF cells were remarkably higher than the control group, and the number of viable cells were not affected by increasing the L-dopa concentration in the range of the study. These results suggest that L-dopa can also have a direct local effect on the growth of HPLF cells. These results may be of use clinically to increase the viability of HPLF cells in cases of an avulsed tooth preserved in storing medium.

Increasing the ectopic uterine motility is the major reason for primary dysmenorrhea. This motility is the basis for several symptoms including for pain is the main complaints of patients with primary dysmenorrhea. There are several mechanisms, which initiate dysmenorrhea. Therefore, different compounds can be employed to control its symptoms. In long-term therapy, combination of oestrogens and progestins may be useful. In short-term therapy, dysmenorrhea sometimes non-steroidal anti-inflammatory drugs (NSAIDs) are used. Most of NSAIDs in long-term therapy show severe adverse effects. In an attempt to find agents with less adverse effect the fennel essential oil (FEO) was chosen for this investigation. In this article, effects of FEO on the uterine contraction and estimation of LD(50) in rat were described. For assessment of pharmacological effects on the isolated rat uterus, oxytocin (0.1, 1 and 10 mu/ml) and prostaglandin E(2) (PGE(2)) (5x10(-5) M) were employed to induce muscle contraction. Administration of different doses of FEO reduced the intensity of oxytocin and PGE(2) induced contractions significantly (25 and 50 microg/ml for oxytocin and 10 and 20 microg/ml PGE(2), respectively). FEO also reduced the frequency of contractions induced by PGE(2) but not with oxytocin. LD(50) of FEO was obtained in the female rats by using moving average method. The estimated LD(50) was 1326 mg/kg. No obvious damage was observed in the vital organs of the dead animals.

Intra-uterine contraceptive devices are associated with an increased incidence of pelvic infections, possible due to the introduction of vaginal bacteria into the uterus at insertion. One potential means to overcome this problem is the use of a device which releases the antimicrobial agent chlorhexidine although such an approach carries with it the risk of adverse effects on the endometrium and, possibly, teratogenic effects. Cultured monolayers of endometrial cells were used to assess the cytotoxicity of both chlorhexidine and chlorhexidine-releasing devices. The results indicated that the agent is toxic at concentrations of 1 microg mL(-1) and that the devices potentiated the toxicity. When the devices were tested in a guinea-pig model, endometrial damage was seen only at the high dose of chlorhexidine, suggesting that there is greater distribution of chlorhexidine in-vivo. Assessment of the teratogenic effects of chlorhexidine in rat embryonic limb bud tissue cells in-vitro showed that the foetal cells were highly susceptible to the toxic effects of chlorhexidine, but that there was no evidence of teratogenicity. Overall, the findings suggest that chlorhexidine-releasing devices may be a safe means of reducing infections related to intra-uterine devices, but that the chlorhexidine may have a toxic effect on foetal cells.

Direct delivery of progestogens to the uterus may be of use in the treatment of dysfunctional uterine bleeding and the sequelae of the menopause as it has the potential to overcome the problems of systemic administration. This study characterised the release of norethisterone and levonorgestrel into an aqueous medium from hollow nylon fibres with dimensions suitable for easy insertion through the post-menopausal cervix. Cell culture techniques were used to assess potential cytotoxic and teratogenic effects. The results demonstrated that the hollow fibres released norethisterone and levonorgestrel at mean rates of 0.5 and 0.6 micrograms/day over 14 days, respectively. There were indications, however, that both steroids were toxic to endometrial cells at concentrations of approximately 5 micrograms/ml, and both drugs showed signs of potential teratogenicity at 10 micrograms/ml. Delivery of the same doses of norethisterone using the hollow fibres reduced the effects on the endometrial and fetal cells. Delivery of levonorgestrel from the hollow fibres had no effect on the endometrial cell toxicity but potentiated the effects on the fetal cells. These results suggest that the hollow nylon fibres may be of use for the delivery of norethisterone to the uterus but that they are inappropriate for the delivery of levonorgestrel.