Positive Control Value Research Articles

Potential of Oyster Mushroom (Pleurotus ostreatus) secondary metabolites on KRAS protein in-silico liver cancer

Liver Cell Carcinoma (KSH) currently ranks as the fourth most common cause of death due to cancer worldwide and is the sixth most common cancer in the world. HCC (Hepatocellular Carcinoma) is caused by environmental factors, viral infections, and certain genetic changes. Ras is a very important oncogene and is hypermutated in various types of cancer, including HCC. The doxorubicin compound is a curative agent that is widely used for chemotherapy treatment of various types of cancer, however, administration of the kura agent with high cytotoxic activity can cause several side effects. Secondary metabolite compounds have anti-oxidant capabilities including breaking free radical chains and chelating redox-active metal ions which cause lipid peroxidation. These properties also help prevent cancer. Pleurotus otstreatus bioactive compounds that have antioxidant potential, flavonol. Due to the potential cytotoxic activity of these secondary metabolites, a bioinformatics approach is needed in this research to determine the results of predicting the interaction activity of flavonol compounds on the KRAS protein in silico and to determine the amino acids and bonds involved in the interaction. between these compounds and the KRAS protein, as a reference for future HCC drugs. The method used is using the PyRx device, Biovia Discovery Studio, Autodock for the docking process and validating the docking results. The results of this research are that flavonol compounds have a docking value of -7.1 kcal/mol. This compound has the potential as a candidate compound that can be used in liver cancer therapy, because it’s have high values ​​and close to the positive control value, namely doxorubicin, of -7.6 kcal/mol.

Effects of Amount and Profile of Amino Acids Supply on Lactation Performance, Mammary Gland Metabolism, and Nitrogen Efficiency in Holstein Dairy Cows.

To evaluate the effects of amount and profile of amino acid (AA) on milk protein yield (MPY), mammary metabolism, and efficiency of nitrogen use (ENU), ten cows were used in 5 × 5 replicated Latin squares and fed a positive control (16.1% crude protein-CP) or two lower CP diets (14.6 and 13.2%) with or without essential AA (EAA) infusion. The EAA solutions provided predicted limiting EAA in each treatment and were continuously infused into the abomasum of the cows. Milk production and MPY were not affected by treatment (mean 35.4 kg/d and 1.03 kg/d, respectively). Efficiency of nitrogen utilization was increased as dietary CP decreased but was not affected by EAA infusion (p < 0.01). Energy-corrected milk production was increased by EAA infusion into 13.2% CP, but not into 14.6% CP diet (p = 0.09), reaching the positive control value. Infusions increased mammary affinity for non-infused EAA (Ile, Phe, Thr, and Trp), allowing the same MPY despite lower arterial concentrations of these AA. Higher arterial concentrations of infused EAA did not increase their mammary uptake and MPY (p = 0.40; p = 0.85). Mammary metabolism did not fully explain changes in N efficiency, suggesting that it might be driven by less extramammary catabolism as AA supply was reduced.

Detection of antibodies to Toxoplasma gondii and Neospora caninum in animals from three zoos in Slovakia.

The aim of this study was to detect antibodies to Toxoplasma gondii and Neospora caninum in exotic animal species kept in three zoos in Slovakia. Antibodies to T. gondii and N. caninum were detected by commercial Enzyme-linked immunosorbent assay (ELISA, ID Screen Toxoplasmosis Indirect Multispecies and ID Screen Neospora caninum Indirect Multispecies, ID Vet, Montpellier, France). Antibodies to T. gondii and N. caninum were detected in 43% (24/56) and 5% (3/55) of animals, respectively. The three animals with N. caninum antibodies: two wolves (Canis lupus) and one Hartmann's mountain Zebra (Equus zebra hartmannae), were clinically healthy, and both wolves simultaneously had antibodies to T. gondii. The results of our study provide a picture of the recent circulation of T. gondii in three Slovakian zoos with the S/P (ratio of antibodies in the sample to antibodies in positive control) value higher than 200%, found in five animals (9%) indicating acute toxoplasmosis. The highest S/P value (296%) was detected in a Roloway monkey (Cercopithecus roloway), which was healthy without clinical signs, presuming that Roloway monkey is a species less susceptible to T. gondii infection. Results of our study showed the presence of T. gondii and N. caninum in Slovakian zoos, confirming recent T. gondii infections according to the high level of antibodies detected in five animals, referring to acute toxoplasmosis.

Analysis of Molecular Docking Chemical Content of Lime (Citrus aurantiifolia (Christm.) Swingle) Against Diabetes Mellitus Therapy Targets and Prediction of Pharmacokinetic Profiles and Toxicity

Diabetes mellitus drugs currently available are sulfonylureas, biguanides, thiazolidines, and alpha glucoside inhibitors which are widely used to control hyperglycemia. These drugs cannot prevent complications of diabetes, and these drugs should not be used continuously because it causes undesirable pathological conditions. The essential oil in lime peel is rich in phenolics, especially flavonoids, which can prevent oxidative stress. This research was carried out computationally to determine the affinity of the compound mechanism in lime, pharmacokinetic profile, and toxicity of the chemical content of lime which is thought to have antihyperglycemic activity using chemoinformatics studies. The hardware used is an Asus laptop X441UB-GA502T Intel Core I5 – 8250 DDR 4 4GB HD 1TB VGA mix 110 2GB screen 14 "DVD-RW WIN 10 ORI. The software used is PLANTS (PROTEIN-LIGAND ANT SYSTEM), YASARA, Marvin sketch, Swisstargetprediction, SwissADME, and Toxtree. All compounds in lime were most active against the target protein PPARγ with an average value of -78.0092, while the positive control value of thiazolidinediones was -90.3393. The highest inhibitory affinity of the compound contained in lime was hesperidin with the target protein DPP-4 of -113.614, higher than the positive control sitagliptin with an inhibitory affinity value of -107.591. Hesperidin absorption in the digestive tract is low, the topology polar surface area (TPSA) value is 234.29Å2, and low polarity and high lipophilicity. There are unexpected heterocyclic compounds, so it becomes a warning against the potential for genotoxic carcinogenicity, namely oxygen element "o".

IN VIVO ANTIDIABETIC SCREENING OF KABAU (ARCHIDENDRON BUBALINUM (JACK) I. C NIELSEN) SEEDS

Objective: Study described the screening potential antidiabetic activity of kabau seed extract and fraction.&#x0D; Methods: The powdered crude drugs weighing 1349.32 grams were extracted with a solvent with solvents with escalating polarity by using soxhletation. The solvents used were n-hexane, ethyl acetate, and 96% ethanol. Screening activity using three variations on doses on the three extracts using the glucose test tolerance method, then the alloxan induction and high-fat feed induction testing methods using selected doses, decreasing blood glucose levels using the GOD PAP enzyme and decreasing MDA levels and increased levels of the enzyme SOD. Extracts that have potential antidiabetic activity are fractionated using liquid-solid fractionation; then the fraction is screened for antidiabetic activity using the glucose test tolerance method.&#x0D; Results: Screening for antidiabetic activity on the three extracts using the glucose test tolerance method showed that the ethanol extract at a dose of 250 mg/kg BW. The three extracts were then screened for the next mechanism using the alloxan induction method and high-fat feed induction, the decrease in blood sugar levels by the GOD-PAP method showed a good decrease in the ethanol extract by 202.94±2 mg/dl, the three extracts showed good less significant, in the SOD enzyme method, the ethanol extract gave a good value such as the positive control value. Testing on fraction can decrease in blood sugar; the results showed that the ethanol extract and methanol fraction gave a small AUC 0-150 (32695,3 and 33167,71), where the value was close to the result of the glibenclamide 30238,48.&#x0D; Conclusion: The antidiabetic activity of the extract showed that the ethanol extract was better with the glucose test tolerance method, with alloxan induction animal models and high-fat feed induction. In the methanol fraction derived from 96% ethanol extract, it provides a good reduction in blood sugar levels in the screening method with a glucose test tolerance

English

Spillage of protoscoleces within hydatid fluid during surgery for hydatid cyst is the main reason for its recurrence. Therefore, to inactivate the protoscoleces, various scolicidal substances have been tested. However, novel and more efficient agents are needed owing to several associated complications. This study focused on the effects of green synthetic Silver Nanoparticles (AgNPs) from Zizyphus spina- christi leaves on Echinococcus granulosus protoscoleces. Also, to evaluate the blood compatibility of Ag NPs. The Ag NPs were identified by ultraviolet-visible (UV-Visible) spectrophotometer, X-ray diffraction (XRD), Scanning electron microscopy imaging, and Energy-dispersive X-ray spectroscopy (EDX). Hydatid fluid was aspirated aseptically from cysts of infected sheep liver. The protoscoleces were exposed to Ag NPs at several concentrations. Also, scanning electron microscopy for ultrastructural changes and in vitro erythrocytes lysis was performed. The Ag NPs were spherical; the particles' size reached 50 nm, and presented a surface plasmon peak around 460 nm. The current study's findings indicated the powerful in vitro scolicidal efficacy of the green biosynthesized AgNPs. Several morphological alterations were observed on the protoscoleces by optical and scanning electron microscopy. Lysis of RBCs at different doses of Ag NPs was significantly (P≤0.05) less than the positive control value, thus proposing its biocompatibility. This work suggests that chemicals like polyphenols present in the extract of Z. spina- christi act as reducing and stabilizers agents to create Ag NPs Nevertheless, further investigations are needed to investigate the Ag NPs scolicidial effects in animal models.

3,6-dimethyl ester-α-mangostin Compound Modified from Isolate α-mangostin Garcinia Mangostana Linn

Isolate α-mangostin (1) is a phenolic compound derived from oxygenated and prenylated xanthones and major compounds obtained from the fruit peel of G. mangostana Linn., Which is stated to have inhibitory activity against α-glucosidase enzymes, serves to determine the antidiabetic activity resulting in IC50 value of 29,92 µM, is of the nature moderate to positive control (acarbose) with an IC50 value of 4.55 µM. A modified compound of α-mangostin (1) with acetic anhydride obtained by 3,6-di-methyl ester-α-mangostin (2) derivative showed the inhibitory value of α-glucosidase (IC50 13,89 μM), this value is better than the activity inhibition of α-mangostin (1), but not as active as the positive control value of the acarbose compound. The separation process to obtain α-mangostin isolates from the fruit peel of G. mangostana Linn was obtained by maceration method with ethyl acetate solvent, followed by refraction using a vacuum liquid chromatography (KCV) method over silica gel (Merck 60 G) and eluted using eluent (n-hexane: ethyl acetate) with increasing polarity, to produce as much pure crystal (21.66 g), yield (24%). While the structural characterization of the two compounds was carried out using UV-Vis, IR, HRESIMS, 1H-NMR and 13C-NMR spectroscopic methods, the antidiabetic testing was carried out using the α-glucosidase enzyme inhibition method.

Controllability Analysis and Control Synthesis for the Ribosome Flow Model.

The ribosomal density along different parts of the coding regions of the mRNA molecule affects various fundamental intracellular phenomena including: protein production rates, global ribosome allocation and organismal fitness, ribosomal drop off, co-translational protein folding, mRNA degradation, and more. Thus, regulating translation in order to obtain a desired ribosomal profile along the mRNA molecule is an important biological problem. We study this problem by using a dynamical model for mRNA translation, called the ribosome flow model(RFM). In the RFM, the mRNA molecule is modeled as an ordered chain of $n$ sites. The RFM includes $n$ state-variables describing the ribosomal density profile along the mRNA molecule, and the transition rates from each site to the next are controlled by $n+1$ positive constants. To study the problem of controlling the density profile, we consider some or all of the transition rates as time-varying controls. We consider the following problem: given an initial and a desired ribosomal density profile in the RFM, determine the time-varying values of the transition rates that steer the system to the desired density profile, if they exist. More specifically, we consider two control problems. In the first, all transition rates can be regulated separately, and the goal is to steer the ribosomal density profile and the protein production rate from a given initial value to a desired value. In the second problem, one or more transition rates are jointly regulated by a single scalar control, and the goal is to steer the production rate to a desired value within a certain set of feasible values. In the first case, we show that the system is controllable, i.e., the control is powerful enough to steer the system to any desired value in finite time, and provide simple closed-form expressions for constant positive control functions (or transition rates) that asymptotically steer the system to the desired value. In the second case, we show that the system is controllable, and provide a simple algorithm for determining the constant positive control value that asymptotically steers the system to the desired value. We discuss some of the biological implications of these results.

High prevalence of fasciolosis and evaluation of drug efficacy against Fasciola hepatica in dairy cattle in the Maffra and Bairnsdale districts of Gippsland, Victoria, Australia

Liver fluke (Fasciola hepatica) is a common parasite amongst grazing livestock in the south-eastern region of Australia and is responsible for significant production losses in the beef and dairy industries. Gippsland in Victoria is a major region for dairy production but no fluke prevalence data in livestock has been obtained in this region since the late 1970s prior to the introduction of Triclabendazole (TCBZ). TCBZ resistance is also now widespread in cattle in south east Australia. In this study, we evaluated the prevalence and intensity of liver fluke infections in dairy cattle in Gippsland and assessed the efficacy of TCBZ and other drenches against F. hepatica on one farm. We obtained 30 individual faecal samples from each of 15 different farms and, using the liver fluke coproantigen ELISA, tested bulk faecal samples pooled from each farm. Any farm that returned a positive bulk sample had all of the samples tested individually to assess the intra-herd prevalence. One farm in the Maffra district also had a coproantigen reduction test and faecal egg count reduction test to assess the efficacy of TCBZ, Clorsulon (CLOR) and Oxyclozanide (OXY). The coproantigen ELISA proved to be a highly sensitive test for liver fluke with a high correlation (R2=0.8849) observed between ELISA data from bulk samples and individual samples, suggesting that future larger scale screening on farms for fasciolosis could use the bulk analysis technique. The ELISA data revealed that animals on six of the 15 farms were infected with F. hepatica and the herd prevalence of the infected herds ranged from 47 to 100% (mean 81%) which exceeds the prevalence value for production losses of 25%. The intensity of fluke infection in cattle varied considerably both within and between herds with a proportion of animals exhibiting a positive control value in the coproantigen ELISA of 50–88%. We also confirmed that TCBZ resistance was present on one farm but that CLOR or OXY can be used to remove the adult stage of the TCBZ-resistant parasites. We conclude that fasciolosis is a significant disease and a likely cause of production losses in dairy cattle in the irrigation zones of Gippsland and that TCBZ resistance is a serious threat to fluke control. We suggest that more work needs to be performed in Gippsland to further define the extent of fasciolosis and drug resistance and to ensure that effective chemical and non-chemical methods of fluke control are incorporated on farms in order to improve animal welfare and reduce financial impacts on producers.

Development and evaluation of an indirect enzyme-linked immunosorbent assay for serological detection of Schmallenberg virus antibodies in ruminants using whole virus antigen.

BackgroundIn late 2011, a new Orthobunyavirus of the Simbu serogroup named Schmallenberg virus (SBV) emerged in continental Europe. The virus is transmitted by hematophagous arthropods, with the Culicoides species as, so far known, main vectors. Infection with the virus can cause clinical signs in adult ruminants including diarrhea, fever and reduced milk production. Transplacental infection of the developing fetus can lead to malformations of varying severity. To assess seroprevalence of SBV in Sweden an indirect enzyme-linked immunosorbent assay (ELISA) was established in connection with the surveys. Here, we describe the development and evaluation of the indirect ELISA, based on whole virus as the coating antigen and a monoclonal antibody for the detection of antibodies to SBV in ruminant sera. The evaluation includes comparison between the in-house ELISA, virus neutralization test and an indirect commercial ELISA.ResultsThe optimal working dilutions of antigens and conjugate were estimated with checkerboard titrations. Comparative studies, including ROC analyses, were used for the selection of an optimal cut-off (S/P value = sample value as percentage of positive control value). With an estimated S/P value of 15% the whole virus ELISA showed a specificity of 100% and a sensitivity of 99.19% compared to virus neutralization test (VNT) and with a good consistency as shown in reproducibility and variability experiments. Furthermore, the comparison of our whole virus indirect ELISA to an indirect ELISA with a SBV nucleoprotein antigen, demonstrated a higher sensitivity of our test.ConclusionThe indirect whole virus ELISA described in this paper is a readily available test for serological analysis of SBV antibodies. Since this in-house ELISA demonstrates a specificity and sensitivity comparable to virus neutralization test and also shows a higher sensitivity compared to commercially available indirect ELISA, it is a useful alternative for surveillance and screening purposes of SBV.

Prevalence of antibodies to Leptospira hardjo in bulk tank milk from unvaccinated dairy herds in the south-west region of Poland.

Bulk tank milk samples were collected from 309 randomly selected dairy cattle herds from the south-western region of Poland in 2010-2011. Samples were tested for antibodies against Leptospira hardjo using DAS-ELISA. Herd level seroprevalence of antibodies against this serovar was low (3.2%). Sample value related to positive control value (S/P ratio) results were highest in herds with 51-100 and 101-500 animals, being 4.6 and 4.1% respectively. The S/P ratio of positive samples indicated a low percentage of infected animals in positive herds.

Risk factors for high‐risk human papillomavirus infection in unscreened Malian women

To investigate the epidemiology of human papillomavirus (HPV) infection in Malian women, for whom cervical cancer is the most common cancer and the second most common cause of cancer-related mortality. Pilot study of 202 women aged 15-65 to determine the prevalence rate of high-risk HPV infection among unscreened Malian women. Information on risk factors was collected through a standardized, structured interview and clinical examination. High-risk (HR) HPV DNA was detected using signal amplification methods (hybrid capture II). High-risk HPV DNA was detected in 12% of unscreened women, while visual inspection after application of acetic acid and Lugol's iodine (VIA/VILI) identified suspicious abnormalities in 2.5% of unscreened women. Histopathological evaluation of VIA/VILI-positive biopsies revealed no evidence of cervical intraepithelial neoplasia or cervical cancer. The majority of infections occurred among women in the 15-24 year old range. Compared to women who were married or widowed, single women were 3.5 times more likely to be infected with HR HPV. The prevalence of infection with cancer causing types of HPV in this study was 12%. These prevalence estimates are consistent with what has been reported previously for other West African countries.

Anti-tumor and Anti-oxidative Activity of Rosmarinus officinalis in 7, 12 Dimethyl Benz(a) Anthracene Induced Skin Carcinogenesis in Mice

The present investigation was undertaken to explore the antitumor-promoting activity of R.officinalis on 2-stage skin carcinogenesis, induced by a single topical application of 7,12-dimethylbenz(a)anthracene and promoted by repeated treatment of croton oil for 16 weeks in Swiss albino mice. Oral administration of R.officinalis leaves extract, during the peri-and post-initiation stage induced by 7,12dimethylbenz[a]anthracene (DMBA), reduced the tumor burden to 2.6 (positive control value: 5.16); cumulative number of papillomas to 13 (positive control value: 62) and percent incidence of mice bearing papillomas to 41.66 %, (positive control value: 100%). A significant (p<0.001) decreased was observed in Lipid peroxidation level by the administration of ROE. Reduced glutathione (GSH) and total proteins was found to be significantly elevated in liver and skin (p < 0.001) of mice in ROE treated group. Skin and liver of R.officinalis treated group animals showed a significant enhancement in antioxidant enzymes like superoxide dismutase (SOD) and Catalase, when compared with the carcinogen treated control. These studies indicate that R.officinalis could reduce the chemical induced tumor and oxidative stress during skin carcinogenesis.

Modulatory influence of Phyllanthus niruri on oxidative stress, antioxidant defense and chemically induced skin tumors

The present study evaluates the modulatory potential of Phyllanthus niruri on chemically induced skin carcinogenesis, and its influence on oxidative stress and the antioxidant defense system. Oral administration of P. niruri extract (PNE), during peri- (Gr. III), post- (Gr. IV), or peri- and post- (Gr. V) initiational stages of 7,12-dimethylbenz(a) anthracene (DMBA)-croton oil–induced papillomagenesis considerably reduced tumor burden to 4.20, 4.00, and 3.33(positive control value 6.20); cumulative number of papillomas to 21, 16, and 10, respectively, (positive control value 62); and incidence of mice bearing papillomas to 50, 40, and 30%, respectively (positive control value 100%), but significantly increased the average latent period to 10.14, 10.62, and 11.60, and inhibition of tumor multiplicity to 66, 74,and 83%, respectively. Enzyme analysis of skin and liver showed a significant (p ≤ 0.05, ≤ 0.01, ≤ 0.001) elevation in antioxidant parameters such as superoxide dismutase, catalase, glutathione, and vitamin C in PNE-treated groups (Gr. III–V) when compared with the carcinogen-treated control (Gr. II). The elevated level of lipid peroxidation in the carcinogen-treated positive control group was significantly (p ≤ 0.05, ≤ 0.01, ≤ 0.001) inhibited by PNE administration. These results indicate that P. niruri extract has potentiality to reduce skin papillomas by enhancing antioxidant defense system.

Abstract 3745: Detection of various ALK translocations using intragenic differential expression (IDE) in patients with non-small cell lung cancer

Abstract Introduction: Efforts to target EML4-ALK fusions with ALK inhibitors in non-small cell lung cancer (NSCLC) have shown promising results in patients harboring a paracentric inversion on chromosome 2, inv(2)(p21p23). In effect, the clinical utility of these drugs is dependent on the presence of ALK gene activation. Reliable testing for ALK activation by translocation is important for selecting patients for this therapy. Although 11 variants for translocation are known, new variants have recently been reported. The aim of this study was to develop a reliable molecular assay to detect translocation of the ALK gene irrespective of the breakpoint on the EML4 gene or the partner gene. To achieve this, we used intragenic differential expression (IDE) of ALK gene comparing expression levels of the 5′ end with the 3′ end. Methods: ALK IDE was determined by measuring the levels of both 5′ and 3′ transcript regions by quantitative RT-PCR. IDE scores were obtained by calculating the endogenous control (ABL1) normalized differences in 5′ and 3′ ALK levels (IDE = 5′ALK/ABL1 - 3′ALK/ABL1). High IDE score, relative to normal, indicates the presence of ALK rearrangement. Relative expression of ALK (independent of rearrangement) was also established. A subset of samples were also analyzed by fluorescence in situ hybridization (FISH). All study samples were analyzed for direct detection of EML4-ALK by RT-PCR performed in a parallel study. Results: The ALK IDE value of an EML4-ALK fusion-positive cell line (NCI-H2228) was 0.7 whereas it was 0.0 for 2 EML4-ALK-negative NSCLC cell lines (NCI H838 and NCI H1299). The positive control value (0.7) was set as the cutoff to distinguish positive vs. negative ALK rearrangement. Eleven percent (6/56) of the lung cancer tissue samples were IDE positive. Eighty-three percent (5/6) of these positives were confirmed by direct detection of EML4-ALK fusion transcript by RT-PCR, including one specimen harboring the previously undescribed variants 8a and 8b. Five IDE-positive samples and 5 with slightly-to-moderately elevated levels of ALK transcript (3′ ALK &amp;gt; 0.1) were further analyzed by FISH. Of these 10 samples, 80% (8/10) showed ALK rearrangement and/or gene amplification. All samples interpreted as having ALK rearrangements by FISH were also positive by IDE (3/3). Two IDE positive (1 confirmed, 1 unconfirmed by RT-PCR) were interpreted as rearrangement negative by FISH. Conclusions: ALK IDE accurately categorized all FISH-confirmed rearrangements as positive and detected rearrangements in at least 1 other confirmed case not identified by FISH. This method is useful for detection of EML4-ALK rearrangements and may function as a universal molecular assay for determining ALK rearrangements in multiple tumor types. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3745.

Studies on the Anti‐inflammatory, Antinociceptive and Antipyretics activities of extract from Leaves of Labisia pumila

The dichloromethane (DCM) extract of Labisia pumila leaves were investigated for its anti‐inflammatory (acetic acid‐induced writhing and carrageenan‐induced paw oedema test), antinociceptive (hot‐plate and formalin test) and anti‐pyretics (yeast‐induced hyperpyrexia test) effects. The phytochemical screening of the extract was also investigated. In the anti‐inflammatory test, DCM extract (100 mg/kg) inhibited significantly the acetic acid‐induced writhing by 78.34% in relation to positive control (aspirin 100 mg/kg) value 88.93% (p&lt;0.05), while the carrageenan‐induced paw oedema was reduced by 50% in relation to positive control (indomethacin 10 mg/kg) value 38.24% (p&lt;0.05) in the experimental rat model. In the antinociceptive test, the extract (100 mg/kg) significantly reduced the licking time in both the early and late phases of the formalin test by 68.41 and 49.17%, in relation to positive control (morphine 5 mg/kg) value 75.17 and 100% (p&lt;0.01), respectively. The extract (100mg/kg) also showed significant effects on hot plate test by 48.75% (p&lt;0.01). Treatment with DCM extract (100 mg/kg) showed a significant reduction in yeast‐induced pyrexia in the fourth hour by 5.4% in relation to positive control (aspirin 100 mg/kg) value 4.3% (p&lt;0.01). Phytochemical screening of the extract indicated the presence of flavonoids, steroids and saponins. The present study confirmed that extracts from the leaves of L. pumila exhibited anti‐inflammatory, antinociceptive and anti‐pyretic activities and these effects might be due to the observed phytochemicals.

The Study of the Anti‐ulcer and Radical Scavenging Effects of Extracts from the Stem of Coscinium Fenestratum

In this study we evaluated dichloromethane (DCM) extract obtained from C. fenestratum, for its ability to protect the gastric mucosa against injuries caused by necrotizing agents (0.3M HCl/60% EtOH), hypothermic restraint stress, and non‐steroidal anti‐inflammatory drugs (NSAID, indomethacin). Total phenoic content and free radical scavenging effects of the bioactive compounds were also determined. In the HCl/EtOH‐induced gastric‐ ulcer model, DCM extract demonstrated significant inhibition of ulcerative lesion index by 98% in relation to the positive control value 72.93 % (p&lt; 0.05), 81% as compared to the positive control group 78.82% (p&lt;0.05) in the NSAID/indomethacin –induced lesion model and 81.97% compared to positive control 73.77% (p&lt; 0.05) in hypothermic‐restraint stress induced gastric acid ulcer model. The radical scavenging activity of the extract was 21.71% which was closely related to the total phenolic content of 256.67mg GAE/g dry weight. HPLC‐UV‐MS profiling of the bioactive compound in the DCM extract confirmed the presence of polyphenolic compounds. From this result we conclude that this extract possess a significant protective effect against gastric mucosa injuries caused by 0.3M HCl/60% EtOH, hypothermic restraint stress, and NSAID/indomethacin in the experimental rats. These effects may be due to the presence of polyphenolic compounds that provided radical scavenging activity.

Evidence That Heparin Saccharides Promote FGF2 Mitogenesis through Two Distinct Mechanisms

Heparin-like saccharides play an essential role in binding to both fibroblast growth factors (FGF) and their receptors at the cell surface. In this study we prepared a series of heparin oligosaccharides according to their size and sulfation level. We then investigated their affinity for FGF2 and their ability to support FGF2 mitogenesis of heparan sulfate-deficient cells expressing FGFR1c. Tetra- and hexasaccharides bound FGF2, but failed to dimerize the growth factor. Nevertheless, these saccharides promoted FGF2-mediated cell growth. Furthermore, whereas enzymatic removal of the non-reducing end 2-O-sulfate group had little effect on the 1:1 interaction with FGF2, it eliminated the mitogenic activity of these saccharides. This evidence supports the symmetric two-end model of ternary complex formation. In contrast, even at very low concentrations, octasaccharide and larger heparin fragments conferred a potent mitogenic activity that was independent of terminal 2-O-sulfation. This correlated with the ability to dimerize FGF2 in an apparently cooperative manner. This data suggests that potent mitogenic signaling results from heparin-mediated trans-dimerization of FGF2, consistent with the asymmetric model of ternary complex formation. We propose that, depending on saccharide structure, there are different architectures and modes of ternary complex assembly that differ in stability and/or efficiency of transmembrane signaling.

The Actin-depolymerizing Factor Homology and Charged/Helical Domains of Drebrin and mAbp1 Direct Membrane Binding and Localization via Distinct Interactions with Actin

Cytoskeletal dynamics are important for efficient function of the secretory pathway. ADP-ribosylation factor, ARF1, triggers vesicle coat assembly and, in concert with Cdc42, regulates actin polymerization and molecular motor-based motility. Drebrin and mammalian Abp1 (mAbp1) are actin-binding proteins found previously to bind to Golgi membranes in an ARF1-dependent manner in vitro. Despite sharing homology through two shared actin binding domains, drebrin and mAbp1 have different subcellular localization and bind to distinct actin structures on the Golgi apparatus. We find that the actin-depolymerizing factor homology (ADFH) and charged/helical actin binding domains of drebrin and mAbp1 are sufficient for regulated binding to Golgi membranes and subcellular localization. We have used mutant proteins and chimeras between mAbp1 and drebrin to identify motifs that direct targeting. We find that a linker region between the ADFH and charged/helical domains confers Golgi binding properties to mAbp1. mAbp1 binds to a specific actin pool through its ADFH/linker domain that is not bound by drebrin. Drebrin localization to the cell surface was found to involve motifs within the charged/helical domain. Our results indicate that targeting of these proteins is directed through multiple distinct interactions with the actin cytoskeleton. The mechanisms for selective recruitment of mAbp1 and drebrin to Golgi membranes indicate how actin-based structures are able to select specific actin-binding proteins and, thus, carry out multiple different functions within cells.

Inhibition of nitric oxide synthesis causes preterm delivery in the mouse.

The aim of the present study was to investigate whether an inhibitor of nitric oxide (NO) synthesis provokes preterm delivery in a mouse model. ICR (CD-1) mice were injected s.c. with N(G)-nitro-L-arginine methyl esther (L-NAME) at 0, 40, 70 or 100 mg/kg on gestation day (GD) 15.5 and 16. Delivery was considered preterm if it occurred before GD 18. In a satellite study, the potential ability of the NO donor sodium nitroprusside (SNP) to prevent L-NAME-induced preterm delivery was tested. Five hours before the initiation of treatment regimen with L-NAME at 70 mg/kg, mice were implanted s.c. with micro-osmotic pumps infusing SNP at 0 or 10 microg/kg/min continuously for 3 days. Administration of L-NAME evoked preterm delivery. This response was noted in 64 and 60% of animals treated with 70 and 100 mg/kg L-NAME respectively (P < 0.05 versus control value). Infusion with SNP provided complete and significant (P < 0. 05 versus positive control value) protection from L-NAME-initiated preterm delivery. This is the first report to reveal that an inhibitor of NO synthesis initiates preterm delivery in a mouse model.