C-myc Monocular Antibody (McAb) on Gastric Cancer Cells

C-myc Monocular Antibody (McAb) on Gastric Cancer Cells Introduction Gastric cancer (GC) is estimated to be one of the most common and frequent malignant tumor of the digestive system. The incidence and mortality of GC have ranked the second among all tumor diseases worldwide [1-5]. However, it ranks in first place in China[6]. Complete surgical resection is still the standard for all patients with resectable GC. It remains highly problematic for the regional and less common systemic recurrences[7]. Recent improvement in surgical technique, adjuvant chemotherapy and radiotherapy has increased the survival rate of patients with early-stage, but the patients who have advanced GC are difficult to cure. With more and more research of molecular biological mechanisms known by us, molecular targeted therapies including cell growth, cell cycle, apoptosis and invasion have become a popular tumor comprehensive therapy[8]. Some of single-targeted spots are mainly Human epidermal growth factor receptor (HER-1, HER-2), Vascular endothelial growth factor (VEGF), Human epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), Cyclin-dependent kinase inhibitor (CDKI), Human proto-oncogene (c-MET)[9, 10]. However, it needs a huge space to develop the targeted anticancer drugs. An elegant way to accumulate therapeutic agents at the tumor site is their specific antibodies[11]. Oncogenes are well documented to be involved in mediating apoptosis and cell cycle resulting in cancers[12, 13], its activation can play an important role in the progress of cancer. C-myc is an important member of the c-myc family and a master regulator of genes involved in diverse cellular processes in GC[14]. The c-myc is a nuclear transcription factor which centrally regulates cell proliferation, differentiation, cell cycle and apoptosis, once c-myc is activated in vivo or in vitro, it is easy to make the cells far from the normal growth and promote cell malignant transformation to cancer finally[15-17]. It was reported that the expression of c-myc is an important consideration in the biological characteristic of GC [18-20]. The previous studies also have proved that c-myc has tight relation with Brest cancer, lung cancer, colon cancer, hematopoietic cancer [21-24]. Currently few da ta exist on the occurrence of the c-myc McAb targeting against GC. In this study, we assessed the effects of c-mycMcAb on the Balb/e2nu/2nunude mice model of GC and the human gastric cancer SGC-7901 cells, and tried to investigate the function of c-myc McAbfor targeting against GC. Materials and Methods Preparation of c-myc monocular antibody (McAb) All experiments involving animals were approved by the Institutional Animal Care and Use Committee of Renji Hospital Affiliated to Shanghai Jiao Tong University of Medicine. Mice were used in this study from Animal Science Laboratory of Shanghai Jiao Tong University, and all effects were made to minimize distress.Thec-myc proteins prepared in E.coliBL21 were used as immunogens.[U1] Before McAb preparation, the c-myc proteins were mixed withequal volume of complete Freund's adjuvant (CFA). Female Balb/c mice aging from 6-8 weeks [U2]were immunized intraperitoneally with 50 ÃŽ ¼g c-myc proteins (1v:1v) in CFA. The immunization was repeated with the same amount of immunogens[U3] in incomplete Freund's adjuvant (IFA) at 14d. A final immunization was performed with 100ÃŽ ¼g mixture of c-myc proteinsand IFA at 28 d. Then, the blood was drawn from the caudal vein and serum titers were measured by ELISA at 35 d. A booster injection was given intraperitoneally at the antibody titers of 640,000[U4] tested by ELISA at 35 d. Five days after boost, spleen cells were isolated and mixed[U5] with SP2/0 myeloma cells. When the Hybrid cells grew to 50%, the positive clones were collected by ELISA. The hybridomas processed by Silica gel H was inoculated intraperitoneally into unsexed Balb/c mice. Then, the mice were scarified and the ascetic fluid was collected. The McAb was purified and the concentration was determined by bicinchoninic acid assay (BCA) Protein Assay Reagent Kit. Characteristic Identification of c-myc McAb The subtype of purified McAb was determined by antibody chips according to the manufacturer's instructions (Raybitech Company, USA) and antibody titers were measured by ELISA kit (Cistron Biotechnology, Pine Brook, NJ)[25]. The assay was carried out in 96-well polystyrene plates according to the standard procedures [26]. Briefly, c-myc protein (10ÃŽ ¼g) were loaded onto plate in 0.1 M carbonate buffer (PH 9.6) and reacted with McAb at 37oC for 2 h. After washed, the mixture was monitored with horseradish peroxidase (HRP)-conjugated rabbit-anti-mouse IgG (diluted 1:100; Sigma) at 37oC for 1 h. Nonspecific antibody binding sites wereblocked with 2% FCS in blocking reagent for 15min.The OD450vaule was read with a 96-well plate reader. The antigenic specificity of McAb was determined by Western blot. Purified c-myc protein was transferred into E.coil DH5ÃŽ ± and Cells were lysed inice-cold radioimmunoprecipitation buffer (RIPA) for 30 min and centrifuged to collect the supermanant[27]. Cell lysates were blent with 3Ãâ€" loading buffer (6 % SDS, 15 % 2-mercaptoethanol, 30 % glycerol, and 0.3 mg/mL bromphenol blue in 188 Tris-HCl, pH 6.8), heated at 90oC for 10 min, and then separated by 16 % SDS-PAGE. Separated proteins in the gels were electrophoreticaly transferred onto nitrocellulose membrane, boiled in phosphate buffered saline for 4 min, and blocked with 5 % nonfat dry milk for 20 min[28]. After several rinses, the membranes were incubated with c-myc McAb overnight at 4 oC. McAb were detected by HRP-conjugated goat-anti-mouse IgG (50 ÃŽ ¼g/mL) at 22 oC for 1 h. The establishment and treatment of nude mice model of GC Four-to-five-weeks old Balb/c nu/nu mice (body weight was 18 ±1.5g) were purchased for the establishment of nude mice model of GC . The human gastric cancer cell line (SGC-7901) was grown in 10% DMEM (Gibco) supplemented with FCS (100 mL/L), penicillin sodium (100 U/mL) and streptomycin sulfate (100 ÃŽ ¼g/mL), and cultured at a 5% CO2 incubator at 37 oC. Exponentially growing SGC-7901 cells were trypsinized resuspended to make a cell suspension of 2Ãâ€"107 cellls/mL. The nude mouse was injected subcutaneously with the suspension (0.2 mL) into the right and left root. Tumor masses were obvious at 10 d. Mice were randomized into 4 groups including low-dose group, middle-dose group, high-dose group and saline group (10 mice/group). Mice of each treatment group were inoculated intraperitoneally with c-myc McAb weekly (10 mg/kg, 20 mg/kg, 30 mg/kg, respectively), and mice of saline groups were injected with normal saline (0.2 mL) instead. Four weeks after injection, the mice were sacrificed and tumors were examined to calculate the tumor inhibition rates (). Immunohistochemistry (IHC) The procedures of SP immunohistochemical kit (SP kit, Maxim Biotech) were as follows: The tissue of tumors was fixed in 10 % phosphate-buffered Formalin, embedded in paraffin, and sectioned at a thickness of 4ÃŽ ¼m. Tissue sections were deparaffnized, hydrated and washed in PBS. Antigen retrieval was performed by combining the tissue with 10mM citrate buffer (pH 6.0) in a microwave for 10 min. Nonspecific protein bindings of tissues was blocked with 5 % normal sheep serum (NSS) for 10 min. after rinsing in PBS, sections were incubated with c-myc McAb at 4 oC overnight at a dilution of 1:100. Secondary antibody (Carpinteria, goat anti-mouse biotenylated, 1:50 in PBS) was applied at room temperature for 30min after washed, and then HRP-conjugated streptavidin were added. The slides were visualized by diaminobenzidine (DAB) (Dako, Carpinteria, CA, USA) for 5 min and counterstained with hematoxylin for 2 min, terminated, dehydrated, transparentized, sealed and photographed step by step. Negative controls were prepared by replacing primary antibody with PBS. Western blot analysis C-myc McAb (1 ÃŽ ¼g/mL, 2 ÃŽ ¼g/mL, 4 ÃŽ ¼g/mL, respectively) were added into SGC-7901cells andcultured for 24 h, 48 h, 72 h, for blank controls, the SGC-7901cells were omitted and HFE-145 cells were used instead. Cells were collected and lysed in ice-cold RIPA,and then following sections were mainlysimilar with the procedures of the Characteristic Identification of c-myc McAb. MTT (3-(4, 5-dimethylthiazole-2-yl)-2, 5-diphenyl tetrazolium bromide) assay Cells were seeded into 96-well plates (10, 000 cells/well) and cultured at 37 oC in a 5% CO2 incubator after HGC-7901cells and normal gastric cell line HFE-145 were trypsinized. The culture medium was washed with PBS 3 times, and thenc-myc McAb (1 ÃŽ ¼g/mL, 2 ÃŽ ¼g/mL, 4 ÃŽ ¼g/mL,) were added respectively, 20 mL (5 mg/mL) At the indicated time points (1 d, 2 d, 3 d, 4 d, 5 d), each well were added with 20 mL MTT cultured at 37 oC for 4 h. Then 150 ÃŽ ¼L DMSO was added again to stop the reaction after the supernate were dropped, The plate was read on multiwall plate reader (Thermo Fisher, Basingstoke, United Kingdom) at 570nm. . A dose-response curve was plotted for the HGC-7901cells and HFE-145 cells. Cell adhesion assay Before cell adhesion and migration assays, SGC-7901 cells and HFE-145 cells were serum starved in bascal culture medium overnight. In brief,6-well tissue culture plates were coated with 10ÃŽ ¼g/mL fibronectin and 10 ÃŽ ¼g/mL poly-L-lysine overnight, the wells were washed with PBS-T and blocked with 5 % BSA in PBS-T[29]. SGC-7901cells and HFE-145 cells were released with trypsin to prepare of single cell suspensions. The cells were applied to 6-well tissue culture plates (50 ÃŽ ¼L/well) and incubated at 37oC for 12 h. When cells were grown to approximately 90 % confluence, the c-myc McAb(1 ÃŽ ¼g/mL, 2 ÃŽ ¼g/mL, 4 ÃŽ ¼g/mL,) were added respectively. Cells were allowed to attach for 2 h, and the culture medium were discarded. Before released with trypsin, cells were washed twice with PBS and 1mM Ethylene Diamine Tetraacetic Acid (EDTA). The formula of was used to calculate the adhesion rates. Cell migration assay Cell migration assays were performed in transwell bicameral chambers as described[30]. Matrigel (Becton Dickinson Company, Bedford) at a dilution of 1:100 were coated with culture medium without serum at 37oC for 30 min in the apical chamber. Cells were released with trypsin, washed ,and resuspended at a final concentration of 5Ãâ€"105 cell/mL in serum-free bascal culture medium (EBM) containing 0.1 % BSA. The suspension (100 ÃŽ ¼L), which were seeded on the upper chamber, were mixed with c-myc McAb (1 ÃŽ ¼g/mL, 2 ÃŽ ¼g/mL, 4 ÃŽ ¼g/mL, respectively). The lower chamber was filled with 500ÃŽ ¼L mouse embryonic fibroblasts (NIH3T3) which were cultured with RPMI (10mM Hepes, 0.5% BSA, pH 7.4). Migration chambers were incubated at 37oC in 5% CO2 incubator for 72h. After removing stationary cells from the upper side of the membrane with a cotton-tipped swab, migrated cells were fixed and stained with 1% crystal violet. Cells were counted in three fields at a magnification of Ãâ€" 400. Cell apoptosis and cell cycle assays HGC-7901cells and HFE-145 cells were seeded into 6-well plates containing Dulbecco's Modified Eagle's Medium (DMEM) with 10% FCS and cultured at 37 oC in a 5% CO2 incubator overnight. Supernatants were discarded, before c-myc McAb (1 ÃŽ ¼g/mL, 2 ÃŽ ¼g/mL, 4 ÃŽ ¼g/mL,) were added respectively and attached for 72 h. cells were fixed with pre-cooled ethanol (75%) and stained with Propidium Iodide (PI, Sigma) in the dark at room temperature for 15 min , after centrifuged at 1,000 rpm and rinses in PBS several times. The cell cycle and apoptosis rates were analyzed by FACS (Elite ESP, Beckman Coulter, Brea, CA). Statistical analyses Statistical analysis was performed using Software version SPSS11.0. All data was expressed as mean  ± SD (standard deviation). Comparisons were made by Student's t-test and comparisons of parameters were made using one-way analysis of variance among 4 groups. A value of P

A NATIONAL FARMING CRISIS :: essays research papers

  Ã‚  Ã‚  Ã‚  Ã‚  Towards the end of the nineteen twenties and through the nineteen thirties of the twentieth century the United States was struck with the largest economic dilemma; the Great Depression. Throughout the Great Depression president’s Herbert Hoover and Franklin D. Roosevelt tried their hardest at reconstructing the nation’s economy so that it would be able to continue it’s path to becoming the world’s greatest nation ever. However, it was a long and several times unsuccessful road which would come to see more than two decades when traveling down it.   Ã‚  Ã‚  Ã‚  Ã‚  During this great depression the farmers of America were greatly effected, maybe more than any other single group of persons in the nation (Bubble Bursts, 133). What came to be known as â€Å"Hoovervilles† popped up across the country, composed primarily of unavailing farmers and their families. In the central area of the United States is where most farmers were affected as it was made of mostly plains and open dirt roads. It was here in central America that farmers gained their most known name used throughout the Depression, â€Å"okies†. These, simply, were the farmers which harshly suffered during the economic downfall. Through times of terror and hardship, when the nation’s economy bottomed out, America’s farmers were faced with the most complex quandary ever; the national farming crisis of the Great Depression.   Ã‚  Ã‚  Ã‚  Ã‚  It is overt to all persons that when materials are grown or produced for sale that a profit is to be made so that money is made back. Between the years of nineteen fifteen and nineteen-nineteen many farmers in the United States actually prospered very well (Baughman, 89). This acute onset of â€Å"good times† was a direct result of European agriculture being temporarily destroyed by World War I (89). As Europe looked to rebuild domestic agriculture they simultaneously looked at nations afar to ephemerally support them. The United States was one of these nations which was looked to for help. Using the supply and demand theory American farmers increased and expanded their supply of crops in order to meet the short-term demand of Europe. This proved to be detrimental to the farming economy of America just several short years later (89).   Ã‚  Ã‚  Ã‚  Ã‚  Eventually, â€Å"†¦Europe was recovering and beginning to rebuild its agricultural sector, it no longer needed to import huge amounts of farm products from abroad†¦Ã¢â‚¬  (Baughman, 89). It was at this time in the early nineteen twenties that the economy of American farming began to plump downward.

Massai Warriors- National Geographic Report :: essays research papers

In the September 1999, issue of National Geographic Magazine, there is quite an interesting article that has been written by Carol Beckworth and Angela Fisher. It deals with the Masai Warriors of Kenya, and how their culture recognizes an adolescent male that is becoming a man, or entering manhood.   Ã‚  Ã‚  Ã‚  Ã‚   The Masai warriors are a group of semi- nomadic people who live on the border of Kenya and Tanzania. They are a relatively small group, with only about 300,000 people in their culture. They hunt for their food with spears, they live in small homes made out of cow dung, and their most advanced form of technology seems to be the bark shoes that they wear on their feet. They are fairly quiet, subdued people, and they seem to ignore the changing world around them. Their customs greatly differ from the outside world, and many of them would nowadays be called very inhumane and primitive. But these ways are the only ways that they know. But, unfortunately, it may not always be that way.   Ã‚  Ã‚  Ã‚  Ã‚   The Masai culture finds the changing of boy to man to be a very important event in life. It is not something that will just happen on it’s own. It is not something that takes place over the course of a decade, either. It is a very spiritual ritual that occurs over a four-day period. This event is known as Eunoto. It is a very rigorous, very challenging, and almost an inhumane ceremony. Eunoto involves the slaying of a lion, the skinning of a buffalo, sexual intercourse with prepuburtal and uncircumcised young girls, the erection of a new building for each young man involved, and very often, the hysterical trance of a young man, during which he may attempt to slay himself with a spear. The young man being initiated finally ends the ritual by having his hair cut off by his mother. This very important event symbolizes the end of the maternal bond between the two. The morals of this culture seem to leave a little something to be desired, however. Marriage, for instance, doesn’t mean quite what it means here (although in some cases, they are more loyal to their spouses there than people are here). A man may be married to more than one wife there, and sex out of wedlock doesn’t appear to be frowned upon there. Many times before the Eunoto is carried out, the young men (18 – 19 years of age) sleep with 9- 11 year old girls.

Allelopathic Effect of S. Macrophylla on the Growth of V. Radiata Seedlings

Allelopathic effect of S. macrophylla on the growth of V. radiata seedlings Thea Philea I. Mostralesa, Greeny Joy A. Perucho, Rhoshela Vi C. Rendon, John Gregor A. Rono, Emmerson P. Rullog, Riffcord R-Denz M. Tabula Institute of Biology, College of Science, University of the Philippines, Diliman, Quezon City (a Thea Philea I. Mostrales, e-mail: theaphilea. [email  protected] com) ABSTRACT Swietenia macrophylla, or commonly known as the mahogany tree in the Philippines, is an introduced species of the family Meliaceae that is commonly used for lumber and reforestation projects.However, based on studies, it is shown that the mahogany tree has inhibitory effects that affect adjacent growing plants. This is made possible chemically and is referred to generally as allelopathy. This study aimed to determine is S. macrophylla had any adverse effects to the growth of plants in close proximity to it. The researchers selected three mahogany trees and planted six plots of differeing distances with each plot containing ten monggo seedlings.The results obtained indicate that there is no significant difference between the growth of monggo seedlings growing near the mahogany tree to that of monggo seedling growing near the control tree based on the analysis done on the seedlings’ height, weight and survival rate. Results also indicate that distance of the seedlings from the S. macrophylla has no effect on the intensity of inhibition of the growth of the V. radiata.The results obtained can be attributed to factors that include the texture of soil, fine-textures soil having been proven to have a greater retention capacity of allelochemicals than coarse-textures soils. Also, allelopathic interactions include both promontory and inhibitory activities of phenolic allelochemicals and thus using seed germination as a bioassay parameter may be of little value. KEY WORDS: Swietenia macrophylla, allelopathy, growth of monggo seedlings, inhibitory effect of mahogany, introduced species INTRODUCTIONBACKGROUND AND SIGNIFICANCE OF THE STUDY Swietenia macrophylla, commonly known in the Philippines as mahogany, is a member of the order Sapindales, under family Meliaceae. It is a large, fast-growing, semi-evergreen tree, popular for landscaping, especially in North American and Carribean countries, where it has a vulnerable status as provided by CITES (Oldfield 1995), and for its strong wood for lumber. This tree was first introduced in the Philippines as early as 1907, and 1913 as part of the Mt. Makiling forest.It is currently used in the Philippines as a lumber tree and utilized in many reforestation projects. However, studies have shown that however beneficial S. macrophylla may be to the Philippine economy and to its environment, it also displays adverse effects to the Philippine wildlife. As a recently introduced species, the said tree has been unable to produce a natural web of life around it; there are no natural consumers of mahogany in the country, an d as such, the area around the tree will not be populated by native fauna immediately.It is also suspected that mahogany leaves contain allelopathic compounds. In a study conducted by P. Thinley in 2002, it was shown that S. macrophylla leaves inhibited the growth of Pterocarpus indicus. Allelopathic compounds inhibit growth and development of other plants when introduced to them. Allelopathy is the production of a certain plant of such compounds and should not to be confused with competition, which may or may not involve allelopathy. Allelopathy is common in the plant kingdom, spread out in random fashion across orders.Some plants are deemed invasive due to their allelopathic nature and mahogany trees are not exempt. STATEMENT OF THE PROBLEM The study aimed to determine if S. macrophylla can adversely affect the growth of plants in close proximity to it and specifically sought to answer the following questions: 1) Does mahogany affect the growth of newly planted seedlings based on the seedlings’ height and weight? 2) Is there a significant difference between the growth of seedlings within the proximity of the mahogany tree to that of seedlings within the proximity of a different tree? ) Is there a significant difference between the survival rate of the seedlings within the proximity of the mahogany to that of seedlings within the proximity of a different tree? HYPOTHESIS The mahogany tree has no effect on the growth of newly planted seedlings. SCOPE AND LIMITATIONS OF THE STUDY The study aimed to determine the effect of the mahogany tree on the growth of monggo seedlings based solely on the seedlings’ weight and height after a specified amount of time.Other possible factors that might affect the growth of the seedlings like availability of sunlight, presence of possible predators and competition with other plants in the vicinity were not included and was not accounted for in the analysis of the data obtained. MATERIALS & METHODS The researchers used monggo (Vigna radiata) as the subject plant. A frequent model plant for laboratory work in Philippine schools, V. radiata is known to be easily grown, requires little maintenance and is fast-growing, which makes it ideal for an experiment limited to a few weeks.Monggo seeds were allowed to germinate by soaking the seeds overnight. Three mahogany trees located along Beta Way in the University of the Philippines Diliman Campus were chosen based on their proximity to other trees. Since competition with other flora could affect the results, it was made sure that the mahogany trees were at least three meters away from the other trees. They were marked as T1, T2 and T3. An acacia tree, Samanea saman, with similar conditions to the experimental trees, particularly to its proximity to other trees, was chosen as the control tree and was marked as T0.To standardize the direction of planting on each tree, angles of 200 east of north and 200 west of south were used to mark radii of 3m each on the north and south side of the tree, respectively. The plots were cleaned and cleared of grass and other flora. Each radius was divided equally into three segments on each segment 10 monggo seeds were planted with a 10cm-interval per seed. Seeds planted on the north side were labelled as N1, N2 and N3 for segments positioned 1m, 2m and 3m away from the tree respectively.The labelling system was used for the south segments. The seeds were uprooted on the 9th day. Seedlings from the same segment were grouped together. The heights of the seedlings were measured using a ruler from the apex of its leaves to the tip of the roots. To standardize the measurement of the height of the seedlings, the roots were cut off at the point where the taproot has become soft and fibrous. The seedlings were then weighed using a top-loading balance. The averages of the height and weight of each segment were then determined. RESULTS AND DISCUSSIONSwietenia macrophylla is known to have inhibitory effec ts that affect adjacent growing plants. This is made possible chemically and is referred to generally as allelopathy. â€Å"The term allelopathy was coined by Molisch in 1937 to refer to biochemical interactions between all types of plants, including microorganisms traditionally placed in the plant kingdom†(Waller 1987). It is defined by Rice (1984) as any direct or indirect beneficial or harmful effect of one plant, including microorganisms, on the other through release of chemicals to the environment.Phenolics, terpenoids, alkaloids, polyacetylenes, fatty acids, steroids and many other different secondary metabolites can act as allechemicals (Rice 1984; Waller 1987; Inderjit et al. 1995). â€Å"However, the mere presence of these chemicals does not establish allelopathy, to demonstrate their involvement in allelopathy, it is important to establish 1)their direct release or indirect origin from plant-derived materials in the environment and 2) that the chemicals are present in sufficient quantities and persist for a sufficient time in soil to affect plant species or microbes† (Putnam & Tang 1986).Allelopathy is different to competition; the latter defined as the removal or reduction of factors from the environment which are vital to survivability, by some other plant or microorganism sharing the same habitat. Allelopathic chemicals, or allelochemicals, are released by higher plants through several mechanisms. These are volatilization, exudation from roots, leaching from leaves or stem by rain,dew or fog, and from decomposition of residues that contain the allelochemicals. Low molecular weight allelochemicals such as those belonging to terpenes are dispersed by volatilization.Nonvolatile allelochemicals such as alkaloids accumulate on plant surface and find their way to the soil by being leached by rainwater. Allelochemicals secreted through root exudation are of several classes. Some are alkaloids, coumarins, flavonoids, and many other types. Al lelochemicals not directly secreted by the plant, in which some pigments are an example, but reach the soil by being released through the decomposition of the plant part that contains them (Leicach et al 2009). In the S. acrophylla, allelochemicals are released by the decomposition of leaf litter. S. macrophylla leaves are a source of tannins, which is evident on the reddish brown color of the dry leaves. Tannins are phenolic compounds that also function as an allelochemical. In February, mature S. macrophyllatrees shed their leaves, which will start to decompose on the ground and consequently release tannins from the cells. Aqueous extracts from the leaves of the S. macrophylla has been shown to retard the growth of Pterocarpus indicuss eedlings (Thinley 2002). Several hydrolysable and condensed tannins were identified as growth and germination inhibitors in dry fruit, growth retarders of nitrogen-fixing bacteria in several plants, and as reducers of seedling growth in several plan ts† (Waller 1987). According to the data and the statistical analyses, the Swietenia macrophylla has no significant effect to the height, weight and survival rate of the Vigna radiate seedlings compared to height, weight and survival rate of the seedlings planted near the Samanea saman. The inhibitory effects exhibited by the S. acrophylla may have been too little or inconsistent to cause a significant change on the growth of the seedlings throughout the duration of the experiment. The compared values of the height, weight and survival rate of seedlings planted near the S. macrophyllaand S. saman are shown in table 1, table 2, and table 3 respectively. The effectiveness of allelochemicals produced by S. macrophylla may have been affected by certain factors. Tannins have been shown to be bound by the humic material I of the soil and presumably inactivated (Waller 1987).The texture of soil has been proved to affect the effectiveness of allelochemicals, favouring fine-textured th an coarse-textured soil, and evidence indicates that the greater retention capacity of fine-textured soils for at least some allelochemicals may be important in the accumulation of physiologically active concentrations of these chemicals (Waller 1987). This may have been the factor that reduced the effects of the allelochemicals involved in our experiment, owing to the beta way’s coarser soil composition. It is also proven that allelochemicals are decomposed in the soil, either abiotically or by microorganisms (Waller 1987).It is also observed that distance of the seedlings from the S. macrophylla has no effect on the intensity of inhibition of the growth of the V. radiata. The effect of distance to the height, weight and survival rate of the seedlings is seen in table 4, table 5 and table 6 respectively. This observation may have resulted from the mechanism in which the allelopathic tannins are dispersed. On the site of the study, the leaf litter covering the soil surroundin g the tree also covered to plots, which meant that the plots may have received variable amounts of allelochemicals as these leached when rains fell.The type of experimentation done was also a factor in investigating the inhibitory effects of allelochemicals. Authors argue whether seed germination is an efficient test in finding out allelopathic potential of phenolic compounds; seed germination is an important parameter for evaluating allelopathic potential of phenolic compounds (Rice, 1984; Waller, 1987), However, using seed germination as a bioassay parameter may be of little value (Stowe, 1979; Inderjit & Dakshini, 1995a). This is because allelopathic interactions include both promontory and inhibitory activities of phenolic allelochemicals.ACKNOWLEDGEMENTS The researchers would like to express their heartfelt gratitude to the following who contributed with the completion of this research: * First of all, to the Almighty God, to whom the researchers dedicate this research study. * To their families and friends who served as their inspirations and the source of very much needed support. * Ms. Lillian Jennifer Rodriguez, for her support and guidance and for lending us pink ribbons for our experiment. * Mr. James LaFrankie, for his contribution to the success of this research study. CONTRIBUTION OF INDIVIDUAL AUTHORSThea Philea Mostrales – Analysis of data, abstract of paper, SP proper Greeny Joy Perucho – Introduction and Methodology of paper, SP proper Rhoshela Vi Rendon – Analysis of data, tables, SP proper John Gregor Rono – Results and discussion of paper, SP proper Emmerson Rullog – Introduction and Methodology, SP proper Riffcord R-Denz Tabula – Results and discussion, SP proper REFERENCES Leicach, S. R. , Sampietro D. A. , Narwal, S. S. , â€Å"Allelochemicals: Role in Plant Environment Interaction†, Studium Press 2009 â€Å"Plant phenolics in allelopathy. † The Botanical Review. New York Botanical Garden. 996. HighBeam Research. 4 April 2013 The IUCN Red List of Threatened Species. International Union for Conservation of Nature and Natural Resources. 4 April 2013 Thinley P. 2002. â€Å"Negative interaction between large leaf mahogany (*Swietenia macrophylla  King) and some indigenous tree secies in lowland forest of Mt. Makiling – allelopathy, a possible cause? †Ã‚  Unpublished B. S. Forestry Thesis, UPLB-CFNR. Waller, G. R. , â€Å"Allelochemicals: Role in Agriculture and Forestry†, American Chemical Society, Washington, D.C. 1987 TABLES Table 1. Independent Samples T-test of the Height of Monggo Plants at 0. 05 significance level | Levene's Test for Equality of Variances| t-test for Equality of Means| | F| Sig. | t| df| Sig. (2-tailed)| Mean Difference| Std. Error Difference| 95% Confidence Interval of the Difference| | | | | | | | | Lower| Upper| height| Equal variances assumed| . 167| . 704| . 737| 4| . 502| 1. 000428290| 1. 357522153| -2. 768657 448| 4. 769514028| | Equal variances not assumed| | | . 737| 3. 900| . 503| 1. 000428290| 1. 357522153| -2. 807242567| 4. 808099147| Table 2.Independent Samples T-test of the Weight of Monggo Plants at 0. 05 significance level | Levene's Test for Equality of Variances| t-test for Equality of Means| | F| Sig. | t| df| Sig. (2-tailed)| Mean Difference| Std. Error Difference| 95% Confidence Interval of the Difference| | | | | | | | | Lower| Upper| weight| Equal variances assumed| . 344| . 589| -. 445| 4| . 679| -. 006665782| . 014968143| -. 048224010| . 034892446| | Equal variances not assumed| | | -. 445| 3. 819| . 680| -. 006665782| . 014968143| -. 049013784| . 035682220| Table 3. Independent Samples T-test of the Survival Rate of Monggo Plants at 0. 5 significance level | Levene's Test for Equality of Variances| t-test for Equality of Means| | F| Sig. | t| df| Sig. (2-tailed)| Mean Difference| Std. Error Difference| 95% Confidence Interval of the Difference| | | | | | | | | Lower| U pper| weight| Equal variances assumed| . 344| . 589| -. 445| 4| . 679| -. 006665782| . 014968143| -. 048224010| . 034892446| | Equal variances not assumed| | | -. 445| 3. 819| . 680| -. 006665782| . 014968143| -. 049013784| . 035682220| Table 4. ANOVA of the Height of Monggo Plants at 0. 05 significance level | Sum of Squares| df| Mean Square| F| Sig. | Between Groups| 14. 889| 2| 7. 44| 4. 598| . 062*| Within Groups| 9. 715| 6| 1. 619| | | Total| 24. 604| 8| | | | *no significant difference Table 5. ANOVA of the Weight of Monggo Plants at 0. 05 significance level | Sum of Squares| df| Mean Square| F| Sig. | Between Groups| . 003| 2| . 001| 1. 960| . 221*| Within Groups| . 005| 6| . 001| | | Total| . 008| 8| | | | *no significant difference Table 6. ANOVA of the Survival Rate of Monggo Plants at 0. 05 significance level | Sum of Squares| df| Mean Square| F| Sig. | Between Groups| . 011| 2| . 005| . 487| . 637*| Within Groups| . 065| 6| . 011| | | Total| . 076| 8| | | | *no significa nt differenceTable 7. Average Height, Weight, and Survival Rate of Monggo Plants Tree| Distance from tree (m)| Height (cm)| Weight (g)| Survival Rate| T0| 1| 16. 67| 0. 2361| 0. 9| | 2| 15. 49| 0. 2347| 0. 95| | 3| 13. 15| 0. 2074| 0. 95| T1| 1| 16. 21| 0. 2255| 1. 0| | 2| 17. 02| 0. 2567| 0. 75| | 3| 16. 73| 0. 2380| 0. 75| T2| 1| 13. 00| 0. 1688| 0. 8| | 2| 18. 99| 0. 2553| 0. 85| | 3| 16. 55| 0. 2340| 1. 0| T3| 1| 14. 22| 0. 1884| 0. 95| | 2| 16. 86| 0. 1928| 0. 9| | 3| 15. 07| 0. 2183| 0. 9| T0 = controlled tree: S. saman T1 = 1st replicate of S. macrophylla T2 =2nd replicate of S. macrophylla T3 = 3rd replicate of S. macrophylla

Accident, Illness and Emergency Policy for Childminders Essay

The safety of children in my care is paramount and it is my policy to take necessary steps to keep children safe when they are in my care. I will promote good health, will prevent accidents and take steps to prevent the spread of illness and infection. I have a responsibility to all children in my care to ensure that a safe and healthy environment is provided at all times. I hold contact details of parents (or another appropriate person, such as grandparent) at all times, provided by parents and stored securely within the child’s records. I hold written permission from each of the children’s parents in the child’s record form files to enable me to seek emergency treatment for their child if it’s needed. As a registered childminder I am required to notify Ofsted of any serious accidents, illnesses or injuries of any child whilst in my care within 14 days of an incident occurring. I will ensure that my pets are safe to be around children and do not pose a health risk. ACCIDENT POLICY As a registered childminder, I am legally required to have a valid first aid certificate. This enables me to administer basic first aid treatment, and my first aid boxes are clearly labelled and easily accessible and are stored in the outside and inside play-room’s. Parents are asked to sign an accident book if injuries occur whilst in my care. Also if a child should come into my care with an injury of any kind, the parent will be asked to sign the book reporting the injury.  Parent/guardian will be contacted immediately. I will use my best judgement as to how other children will be looked after. You would be consulted about this by phone. There is a parent ring round provision in the event of an emergency to inform all parents of children under my care. If an accident or incident involving a child in my care may result in an insurance claim I will contact my public liability insurance provider to discuss my case and be allocated a claim number. This may involve discussing details of the child in my care with a third party. ACCIDENT PROCEDURE In the event of an accident whilst the child is in my care, I will: †¢ deal with any minor accidents. †¢ record all information, including names of witness(es), in the accident book. †¢ request parents, on picking up the child / children, to sign the accident book and read the information regarding the incident. †¢If I am unable to get hold of parents or other named appropriate adult, he/she will immediately be taken to see a doctor at Bishops Waltham Health Centre – or hospital if necessary and I will stay until parent arrives In the event of an accident in the home: †¢ Parents must inform me of any accident at home and record it in the accident at home area of the accident book. Further observation of injury on a child which has not been reported to me by the parent nor occurred whilst in my care, parents will be asked for information on the incident and instructed to write in the back of the accident book. In the event of me being subject to an accident: If I have an accident, I will get the nearest responsible adult to help, while my emergency back-up people are being contacted. I will do my best at all times to make sure the children in my care are safe, reassured and kept calm. ILLNESS POLICY I will do everything I can to take care of your child if they are feeling unwell by comforting them and giving them appropriate medicine you have approved and have signed a declaration permitting me to administer. If I am in need of support or advice regarding a serious illness or incident involving a child in my care I may contact National Childminding Association who will log information regarding the incident with regard to their safeguarding policy. A NCMA designated officer will be assigned to my case and all the information given to them will be kept confidentially unless their appear is to be a child protection issue which will be reported accordingly. ILLNESS PROCEDURE You must inform me if your child has been ill overnight or the previous day. I might have other children I am taking care of and need to ensure your child does not have an infectious illness. Medicines will be kept in a safe place and will be logged in the ‘medicine book’. However, if you child arrives ill, I will take a decision as to whether the child is fit to attend or not. If not, the parent will be asked to take the child home. If your child becomes ill whilst in my care, I will: 1.Contact you for agreement to administer ‘calpol’ (if appropriate and agreed) and if your child remains ill after one hour you will be asked to collect your child and make a doctors appointment. 2.You will be required to keep your child at home until they are recovered so that your child can be cared for on a one to one basis at home. 3.In the following circumstances children may be required to stay at home: Diarrhoea and/or vomiting, a rash, conjunctivitis. 4.If I am unable to get hold of parents or other named appropriate adult, he/she will immediately be taken to see a doctor at Bishops Waltham Health Centre – or hospital if necessary and I will stay until parent arrives Infectious illness: In the event of your child contracting chicken-pox, mumps, measles etc, or a fever or temperature of 101 degrees Fahrenheit, or 38 degrees centigrade or over your child will not be able to attend under my care and must remain at home. EMERGENCY POLICY Parent/guardian will be contacted immediately. In the event of an accident requiring hospital treatment, I will attempt to inform the parents immediately and take the child to hospital. I will stay with the child until the parents arrive. EMERGENCY PROCEDURE As a registered Child-Minder and trained paediatric first aider, I will ensure that the most appropriate treatment is given or acted upon for your child in my care; however if I have more than 1 child in my care, I will use my best judgement as to how other children will be cared for. Initial action will be reporting the incident to the parent by phone. We practice a parent ring round provision in the event of an emergency to inform all parents of children under my care. If parents do not arrive or I am unable to contact them, I will stay with the child until the appropriate treatment has been given and then return home where your child would be cared for until the arrival of the parent

On the feature of American slangs Essay

Abstract: With the development of the society, slang is accepted by more and more Americans. This thesis analyses features and functions of American slang. The author draws a conclusion that American slang reflects the cultural characteristics of Americans’ humor, informality, self-confidence, anti-authority, innovation consciousness and pursuit for time efficiency. Slang is a sort of informal language, the original purpose is to express vulgarly or use it as a jargon or cant. With the development of the modern American culture, the modern American slang has become a common linguistic phenomenon which is different from jargon and argot, spreading all over any aspects of life. In American English, slang is also one of the major parts in people’s life and can reflect the social life about civilian requirements and characteristics of language expression. According to accurate statistics, the daily American vocabulary is about 10000-20000 words, of which 10% about 2000 words are slangs, these slangs are frequently used in people’s daily life, with strong expression and abundant culture feature, therefore, only by having some certain knowledge on American slangs can we speak American English properly and natively. Here is what I am going to discuss on the characteristics of modern American slang, and the translation of American slang as well, so as to improve people’s understanding and the use of American slangs. The basic characteristics of modern American slang (a) informality The most significant feature is informality, which is the origin of the slang and nature of it. Because of the informal expression of American slang, moreover, comparing the American English with British English, the former is actually more flexible and informal. Therefore, it also determines the informality of American slang and makes it even more apparently. For instance: A:Excuse me. I have to catch some Zs. (Excuse me. I have to take a nap.) B:I thought you just woke up. Sleepyhead.(I thought you just woke up. You are lazy and sleepy.) In this example, a typical slang â€Å"catch some Zs† by the speaker means taking a nap, and â€Å"sleepyhead† means a person who loves sleeping, both of which are limited to oral English expression frequently, generally not used for formal expressions. In addition, common American slang expressions such as â€Å"kiss ass (sycophancy)†, â€Å"hit the road (set out)†, and â€Å"take a dump (defecate)† are typically informal. It is because of its obvious informality that Americans like advocating freedom of expressions in some light-hearted situations that need humorous atmosphere to ease embarrassing scene. Both politicians or ordinary people prefer using slangs to light up the atmosphere. (b) creativity America is a country that desires for freedom. In pursuit of innovation, people are unremitting to new things and new ideas, which creates a large number of novel American slang words and provide a perfect condition. Furthermore, one of the characteristics of American slang is innovative. American slangs always use the original or new words to reflect new things, new development and new concept. Like the word â€Å"jazz† , it was first used to express a kind of music in New Orleans. Along with the development of jazz, Americans soon produced â€Å"bop† (pop), â€Å"far – out† (fashion), â€Å"beat† , â€Å"cool† (modern), etc. Because of the style of jazz, the word â€Å"jazz† generates some new meanings, such as â€Å"exertion†, â€Å"cordiality†, â€Å"frenzy† and so on. The old American drug addicts created the slang nose candy, black stuff, shooting gallery referring to drug. The blacks create and use slang a lot, such as dog juices (cheap wine), cakewalk (a kind of hip-hop dance), breakdancing (break), etc. Since the 1980s, some modern terms have been popular among the young, such as â€Å"bag your faith (left)†, â€Å"big time (serious)†, â€Å"golden (women)†, â€Å"worms (ugly people)†. They are the creative use of the phrases from the slangs. The generation of slang is inseparable from the creative use of the original English vocabulary, and the creation of new words plays a vital role as well. The United States is a multi-national country with immigrants, and constantly the moving in immigrants creates new words, making the American slangs increase rapidly. All of these prove that American slang is innovative. (c)universality In American English, the use of slangs are very common. It has something to do with the pursuit of flexibility, innovation, humor in American English, also with the history of United States. In the history of the United States, â€Å"westward movement in the contemporary† formed the largest population flow, so that the regional dialect and the jargon from all classes including cant have been widely spread. Because the official does not emphasize on â€Å"literary language†, and has made few â€Å"language purification†, thus jargon and sayings which were constantly used , have formed today’s omnifarious slangs that Americans generally use. In the 20th century, American slangs had a development which progressed by leaps and bounds. During the two world wars, the Vietnam war, the Korean war, and the cold war period, a large quantity of slangs generated. Moreover, American social problems, such as racial discrimination, civil rights movement, family disintegr ation, sexual liberation, gay so on and so forth, much more American slangs have mushroomed. English slangs can be defined to common colloquial language, and according to the definition of the Concise Oxford English Dictionary (1985), slangs are frequently used as colloquial words and phrases. It cannot be taken as Standard-English, therefore this demonstrates that, slang is Nonstandard-English. Nevertheless, slangs are very popular among the English-speaking countries nowadays. Slangs are spoken by people in daily speech, and even magazine and literary works may find a large number of them. With the development of the society, slangs are widely used by more and more people, especially in America, slang can be found favorable in the Americans’ eyes. It is accepted by both upper-class and lower-class, and especially the college students who tend to use up-to-date words that form a kind of characteristic style of themselves. Slang plays an important part in American English. It is reported that slang accounts for one of the tenth of American average vocabulary. People’s attitude toward slang varies from person to person. Some think that slang cannot be used on the formal occasion, for it is spoken by the illiterate people only and the slang terms are so coarse. Some even think that slang is the filthy language, and advocate abandoning using it. However, others hold the opposite idea. They think that slang play an important role in English language, because of its fresh, lifelike, humor and adequate expressions. One must admit the undeniable fact that slang is spoken by more and more people. In modern times, if one is not familiar with American slang he or she could not capture the slang expressions in the movie and common fictions, and, what worse, one would 0make mistakes when they are talking with the Americans. An example will clarify the fact. A guide asks the tourist â€Å"anybody wants to take a leak?†(Does anybody want to go to the restroom?), and, of course, what the guide said would make the tourist surprise. It is the uneducated vulgarity of expression at all. Therefore, one should bear in mind that it is very important to study language in a sociocultural context; the language has existed in close contact with culture. Fro m the example one can know that misusing slang would result in misunderstanding in the communication. It can not tell others that one is good at English. It is not thus; moreover it shows that one just has half-back knowledge about slang. Americans speak colorful, idiomatic English that may have little resemblance to the language one studied in school. Even a native English speaker might be confused by the combination of slang that characterizes American conversation. So it is very important to capture the American slang expression in order to know others correctly. This thesis starts with the analysis of the cultural condition of American slang, and its purpose is to probe into the features of slang so as to bring the relationship between slang and society culture to light. â…  .American Slang is quite a controversial language, people have different attitude toward it. It is no wonder that the definition of slang is so variety. However, the slang terms have their own characters, and, of course, the definitions of it are more or less described in the same way. A. The Definition of Slang Slang is a rather special parole. It is neither the word that beyond the regular role of grammatical usage, nor rural dialect. Although it is quite different from acrolect which used on the formal occasion, people quite like to use it in daily speech. As a matter of fact, the meaning of most slang is similar to the words of colloquial style. Some consider one word is slang, while others might not think so. For example, the old may believe the word bike is slang, but the youth might not .Bicycle is trisyllabic, while bike is monosyllabic, it is so troublesome to pronounce the word with trisyllabic, and of course, the youth prefer to omit some other two syllables, and bike is spoken by the youth more often. To some certain slang, it is hard to tell the differences between slang and colloquial style. One of the definitions of it is, â€Å"the body of words and expression frequently used by or intelligible to a rather large portion of the general public, but not accepted as good; formal usage by the majority† (Flexner, 1975:3).

Assignment african american history Essay Example | Topics and Well Written Essays - 1500 words

Assignment african american history - Essay Example Although most people today believe slavery in America was confined to the south, Douglass witnessed many slaves being beaten during his early childhood in Maryland. He was often required to endure cold and hunger due to neglectful conditions in his northern home. When he was eight years old, he was sent to work for a ship’s carpenter in Baltimore. While there, he learned to read and write until his mistress was informed this was against the law. His experience in the city made him aware that not everyone bought into the idea of slavery. When Douglass was 15, his owner died and he was sent back to the farms. There, he was cruelly beaten by the slave-breaker Edward Covey until the day Douglass beat up Covey and tried to escape. He was caught and returned to slavery, but sent back to Baltimore. In Baltimore, Douglass borrowed the identification papers of a free sailor friend and successfully escaped on September 3, 1838. He began writing about his experience in 1845. Douglass’s narrative reveals the degree to which black people were made into beasts. Although his mother died when he was seven, he admits the news had almost no effect on him. This was because he had been separated from her since infancy. â€Å"Never having enjoyed, to any considerable extent, her soothing presence, her tender and watchful care, I received the tidings of [my mother’s] death with much the same emotions I should have probably felt at the death of a stranger† (Ch. 1). At this point, he’d also watched his aunt brutally whipped and he was working in the fields. Early separation from family destroyed any natural human feelings of attachment and removed any possible support. Cruel treatment kept him always in fear. Neglectful living conditions made him grateful for the smallest crust of bread. His description of the life of the slave reveals the need for and active encouragement of bestial