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Jornal Brasileiro de Pneumologia
Print version ISSN 1806-3713
J. bras. pneumol. vol.38 no.5 São Paulo Sept./Oct. 2012
Proposed short-term model of acute allergic response, without adjuvant use, in the lungs of mice*
Andrea Mendonça RodrigueI; Camila Zanelatto Parreira SchmidtII; Lucien Peroni GualdiIII; Raquel Giacomelli CaoIV; Rodrigo Godinho de SouzaV; Ana Cláudia PereiraVI; Nailê Karine NuñezVII; Alisson Passos SchleichVIII; Paulo Márcio Condessa PitrezIX
IDoctoral Student. Graduate Program in Pediatrics and Child Health, Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
IIPediatric Pulmonologist. Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
IIIDoctoral Student. Graduate Program (Joint Doctoral Program) in Pediatrics and Child Health, Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
IVDoctoral Student. Graduate Program (Joint Doctoral Program) in Pediatrics and Child Heath, Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
VTechnician. Laboratory of Pediatric Pulmonology, Biomedical Research Institute, Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
VIBiology Student. Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
VIIMaster's Student. Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
VIIIBiology Student. Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil
IXProfessor. Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - School of Medicine, Porto Alegre, Brazil
OBJECTIVE: To determine whether a short-term protocol using subcutaneous sensitization with ovalbumin, without the use of adjuvants, would induce an eosinophilic response in the lungs of mice similar to that observed in previous, well-established protocols.
METHODS: Adult female BALB/c mice were randomized and divided into groups according to the number of sensitizations with ovalbumin and the number/dosage of intranasal ovalbumin challenges. The short-term protocol (10 days) consisted of one sensitization with ovalbumin and three ovalbumin challenges (100 µg). Total and differential cell counts in BAL fluid, levels of eosinophil peroxidase in lung tissue, and histopathological examination of the lungs were performed 24 h after the last ovalbumin challenge.
RESULTS: No significant differences were found among the groups regarding the variables studied. The short-term protocol, as well as the other protocols studied, induced an eosinophilic response similar to that obtained in the positive control.
CONCLUSIONS: Subcutaneous sensitization with ovalbumin and without the use of adjuvants resulted in a significant allergic response in the lungs of mice, even in the short-term protocol group. Our findings suggest that this short-term protocol can be used as a first-line pre-clinical test for the study of new medications, reducing the costs and observation periods.
Keywords: Ovalbumin; Mice; Asthma.
Asthma is a chronic lower airway disease that affects approximately 300 million people worldwide, resulting in high morbidity and substantial costs to society. Asthma is characterized by infiltration of CD4+ Th2 cells and eosinophils in the airways, associated with production of allergen-specific IgE.(1) In the study of new therapies for asthma, as well as for a better understanding of the pathophysiology of the disease, animal models have been used for decades.(2) Murine models involving systemic sensitization to antigens, with subsequent airway challenge, have been developed in order to trigger an allergic response in the lungs similar to that of asthma.(3) These models are still the most commonly used in experimental asthma research, despite some recent criticisms of factors related to current animal models of asthma, which do not develop phenotypic characteristics similar to those of asthma in humans.(4)
Among the animals studied, isogenic mice are the most popular, because they are inexpensive, there is detailed knowledge of their genetics, and there is a clear Th2 immune response in their lungs when they are exposed to specific allergens.(3) Classically, BALB/c mice are intraperitoneally sensitized with ovalbumin twice, 14 days apart, together with an adjuvant, i.e., aluminum hydroxide (alum); although this method is routinely used, it is an artificial way of boosting the allergic response in the lungs.(5) In the literature, this has been the standard protocol most commonly used as a murine model of asthma. Overcoming two frequently reported limitations (use of adjuvants and prolonged protocol duration) would be attractive for performing murine models of asthma that are more sophisticated, especially in pre-clinical testing of new drugs.
In an attempt to find an alternative to the use of adjuvants in murine models of asthma, Conrad et al. have recently demonstrated that subcutaneous sensitization with ovalbumin, without the use of adjuvants, results in an allergic response in the lungs similar to that observed in previously, well-established protocols.(6,7) The results of that study allow the exclusion of the use of adjuvants in murine models of asthma in future studies; the artificial way in which asthma is currently studied in animal models, with the use of adjuvants, has been justifiably criticized. However, in that study, there were three sensitizations, 7 days apart, and the protocol remained too long. To date, there have been no studies testing short-term protocols for use in murine models of asthma, particularly without the use adjuvants. If proven viable, this type of model would be an interesting means to evaluate, preliminarily, new therapies for asthma, particularly those related to an eosinophilic response in the lungs, such a response being a key outcome in therapeutic targets of atopic asthma. In the pharmaceutical industry setting, long-term protocols have been a significant limitation in the study of new drugs.(8)
Therefore, the objective of the present study was to find alternative protocols for animal models of asthma, i.e., protocols that are shorter and less artificial. A 10-day protocol with only one subcutaneous sensitization with ovalbumin and without the use of adjuvants was compared with standard protocols described in previous studies.
A total of 44 isogenic BALB/c mice (adult females 6-8 weeks of age) were obtained from the Rio Grande do Sul State Foundation for Health Science Research. The animals were fed a balanced chow diet and had ad libitum access to water, being housed in cages and maintained on a 12/12-h light/dark cycle. The animals were divided into the following groups:
five study groups of 7 animals each, in which different models of induction of an allergic response in the lungs were tested
one positive control (C+) group of 5 animals
one negative control (C-) group of 4 animals
The groups were classified according to the type of sensitization (intraperitoneal or subcutaneous sensitization with ovalbumin, with or without the use of adjuvants), the number of sensitizations (one or two), the number of intranasal ovalbumin challenges (two or three), the dosage of intranasal ovalbumin challenge (40 µg or 100 µg), and the observation period (21 or 10 days). The control groups were observed for 28 days.
Sensitization with ovalbumin and intranasal ovalbumin challenge were performed with total volumes of 200 µL and 50 µL, respectively, diluted in Dulbecco's PBS (DPBS). The dose of ovalbumin used for sensitization was 20 µg in all groups except the C- group. For intranasal ovalbumin challenge, the animals were anesthetized with isoflurane in an anesthesia chamber to allow pulmonary aspiration.
The animals in the study groups underwent the following:
2sc2in-100 group-two subcutaneous sensitizations with ovalbumin (days 0 and 7) and two intranasal ovalbumin challenges (100 µg) on consecutive days (days 19 and 20)
2sc3in-100 group-two subcutaneous sensitizations with ovalbumin (days 0 and 7) and three intranasal ovalbumin challenges (100 µg) on consecutive days (days 18, 19, and 20)
2sc3in-40 group-two subcutaneous sensitizations with ovalbumin (days 0 and 7) and three intranasal ovalbumin challenges (40 µg) on consecutive days (days 18, 19, and 20)
2sc2in-40 group-two subcutaneous sensitizations with ovalbumin (days 0 and 7) and two intranasal ovalbumin challenges (40 µg) on consecutive days (days 19 and 20)
1sc3in-100 group (with a short observation period, which was the primary objective of the present study)-one subcutaneous sensitization with ovalbumin (day 0) and three intranasal ovalbumin challenges (100 µg) on consecutive days (days 7, 8, and 9)
The first four groups were observed for 21 days, whereas the 1sc3in-100 group was observed for 10 days (short-term protocol).
The animals in the control groups underwent the following:
C+ group: two intraperitoneal sensitizations with 1 mg of ovalbumin + alum (days 0 and 14) and three intranasal ovalbumin challenges (100 µg) on consecutive days (days 25, 26, and 27)
C- group: two subcutaneous administrations of DPBS (days 0 and 14) and three intranasal instillations of DPBS on consecutive days (days 25, 26, and 27)
The protocols are shown in Figure 1.
For BAL, the animals were anesthetized with a solution of xylazine (100 mg/mL) and ketamine (100 mg/mL), at a ratio of 1:9 (dose, 0.1 mL), and the trachea was cannulated with a blunt needle. A DPBS solution (1 mL) was instilled intratracheally and immediately aspirated. This procedure was performed three consecutive times. After the BAL procedure, the animals were euthanized with lethal doses of the drugs used for anesthesia (dose, 0.3 mL i.p.) and were disposed of in accordance with the norms of the institution.
The BAL fluid was weighed and centrifuged at 1,500 rpm for 10 min at 4ºC. The precipitate was resuspended in 1 mL of DPBS. For all samples, total cell count and cell viability in the suspension were determined by the trypan blue exclusion test with a Neubauer chamber (BOECO, Hamburg, Germany). For the differential cytologic analysis, the precipitate suspension (30 g) was placed in a cytospin centrifuge for 5 min. The cells were analyzed for their morphology by the May-Grünwald-Giemsa method. The cell types observed under an optical microscope are expressed as percentages, after the counting of 200 cells.
After BAL and lung resection, we used ortho-phenylenediamine (a chromogenic substrate) in order to produce a chemical reaction and measure eosinophil activity in lung tissue. As demonstrated by Strath et al.,(9) this method allows us to quantify eosinophil activity on the basis of the eosinophil peroxidase (EPO) reaction, by measuring optical absorbance, without interference from other peroxidases that might be present in the tissue analyzed. In brief, lung tissue fragments were frozen and thawed three times in liquid nitrogen. After centrifugation for 10 min at 4ºC, the supernatant was serially diluted five times in 96-well plates (50 µL/well). Subsequently, 100 mL of substrate (1.5 mM ortho-phenylenediamine and 6.6 mM hydrogen peroxide diluted in 0.05 M Tris-HCl buffer, pH 8.0) were added. After 30 min, at room temperature, the reaction was stopped by the addition of 1 M sulfuric acid, and the absorbance of the samples was measured at 492 nm.
After having been removed, the lungs were perfused with 10% buffered formalin on a gravity column (20 mmHg). The specimens were embedded in paraffin blocks, cut into 4-µm sections, stained with H&E, and mounted on slides. In order to determine the severity of eosinophilic inflammation, we performed a qualitative assessment of bronchial inflammatory response under an optical microscope, by counting the number of eosinophils in a given field, in three different bronchi.
The statistical analysis was performed with the Statistical Package for the Social Sciences, version 17.0 (SPSS Inc., Chicago, IL, USA). After descriptive analysis of the variables, we used the Kruskal-Wallis test with Dunn's post hoc test for comparisons that reached statistical significance. The C- group was statistically tested separately for each study group by the Mann-Whitney test, with adjustment for multiple comparisons. The level of statistical significance was set at 0.05. The eosinophil count in BAL fluid was used as the primary outcome measure because eosinophils are the most important immune effector cells in this experimental model.
The sample size was calculated at 7 animals per study group. This calculation was based on the following: absolute eosinophil count in BAL fluid for a mean of 0.7 × 106 cells/mL as the primary outcome measure; a standard deviation of 0.34; a value of p = 0.05; and a power of 80%, the difference among the means for the groups having been estimated at 80%.
The present study was approved by the local animal research ethics committee and was conducted in accordance with current ethical norms for animal model research, following the guidelines of the Brazilian Association for Laboratory Animal Science, using as few animals as possible, and managing pain and suffering during all procedures, including euthanasia.
In the present study, models that induce an acute allergic response in the lungs of mice by using subcutaneous sensitization with ovalbumin and different numbers and doses of intranasal ovalbumin challenges were compared with one another and with the traditional model (C+ group). The primary goal of the study was to determine whether the use of a short-term (10-day) protocol consisting of one sensitization would induce an allergic response in the lungs similar to that observed in standard protocols in the literature. The mean BAL fluid return volume for all animals was 0.55 ± 0.15 mL, and the mean cell viability was 100%. The mean eosinophil count in the C- and C+ groups was 2.5 ± 3.0% and 40.1 ± 10.0%, respectively.
There were no significant differences between the study groups and the C+ group regarding total cell count (p = 0.1), lymphocyte count (p = 0.36), macrophage count (p = 0.24), or neutrophil count (p = 0.059) in BAL fluid. The eosinophil count was higher in the 2sc3in-40 group than in the 2sc2in-40 group (p = 0.032; Figure 2). There were no significant differences in cellular inflammatory response in the lungs between the animals undergoing the short-term protocol, without the use of adjuvants, and those undergoing the 21-day protocol that is widely used in animal models of asthma. In comparison with the C- group, the study groups showed significantly higher total and differential cell counts (p < 0.05).
Histological examination revealed intense peribronchovascular inflammatory infiltrates in all groups, with a predominance of lymphocytes and the presence of more than 100 peribronchial eosinophils per field. The anatomy of the animals in the C- group was preserved, without inflammatory changes, a finding that is characteristic of healthy animals. From a histological standpoint, there was no difference in the severity of pulmonary inflammation among the groups, including the group undergoing the short-term protocol (Figure 3). Analysis of EPO levels in lung tissue, which reflect eosinophil activity in the lungs, revealed no differences among the groups studied (Figure 4).
Animal models of asthma with shorter-term protocols and without the use of adjuvants seem to be an attractive alternative for experimental studies of asthma in certain situations. Using the outcome variables that are more directly related to the presence and activity of eosinophils in the lung, we compared different groups of BALB/c mice sensitized with ovalbumin and found no significant differences among the different protocols studied, including a short-term (10-day) protocol.
The most important finding of the present study, especially if we take into consideration that no adjuvants were used, is that there were no differences among the protocols despite their differences in duration (10, 21, or 28 days). We demonstrated for the first time that a shorter-term protocol, without the use of adjuvants, can induce an allergic response in the lungs of mice similar to that observed in previous, well-established protocols. The use of a 10-day protocol inducing the same eosinophilic response in the lungs as that observed in previous, longer-term protocols widely used in the literature can facilitate pre-clinical trials (particularly those investigating new therapeutic targets), reducing the costs and the duration of experiments. In addition, shorter-term protocols are in accordance with all ethical principles currently guiding the use of animals in research, reducing the duration of the experimental procedures that animals undergo. There were no significant differences between the short-term protocol tested in the present study and the protocol used in the C+ group in terms of total cell counts and eosinophil counts in BAL fluid or in terms of EPO levels in lung tissue. In addition, for the short-term protocol, histological examination revealed a significant peribronchovascular inflammatory cell infiltrate, with a predominance of eosinophils, with the same characteristics as those observed for the other protocols.
In 2008, Hahn & Erb published an article questioning the time it takes to identify and develop new treatments for asthma; this process begins with identifying a molecule and determining whether it is actually involved in the disease process, and it is followed by identification of the structural components and synthetic modification of the molecule, culminating in pre-clinical testing in animals.(8) In addition to this long and costly process, there are long periods of experiments in mice. It is of note that the results obtained with the model proposed in the present study are related to the analysis of acute response to a single allergen (i.e., ovalbumin). It is also of note that experimental models of asthma involving chronic exposure have been proposed and are important in the study of new drugs, being probably performed in a later sequence of experiments, when therapeutic targets are sought.(10,11) Shorter-term protocols can nevertheless be an interesting screening model for new therapeutic targets.
In 2004, Cates et al. conducted a study involving a short-term (10-day) protocol of sensitization with house dust mites (HDM) and demonstrated eosinophilic inflammation in the lungs of mice, as well as increased expression of Th2 effector cells (CD3+, CD4+, and T1/ST2+), together with increased levels of total IgE and HDM-specific IgG1.(12) The results of that study and those of our study demonstrate that it is possible to perform a short-term (10-day) protocol using either ovalbumin or HDM.
Various authors have criticized the use of alum in murine models of asthma because alum is an artificial substance for the induction of an immune response, inducing pro-inflammatory cytokine production and activating Th2 lymphocytes.(5,13,14) Alum also activates the immune system via dendritic cell maturation and by co-stimulation of expression molecules.(15,16) In addition, alum has been shown to cause major stress in animals up to 4 h after exposure.(6) Studies in which alum was not used demonstrated that this substance is not necessary for the induction of an allergic response in the lungs of mice and that there is no difference between subcutaneous sensitization without the use of adjuvants and intraperitoneal sensitization with alum. Those studies, involving protocols without the use of adjuvants, demonstrated different levels of inflammation, different levels of immune tolerance to the allergen, or protocols that were often too long.(7,17-20) More recently, Conrad et al. found no significant differences in BAL cellularity, lung histology, or interleukin (IL-5, IL-10, and IL-13) levels using subcutaneous sensitization with ovalbumin and no adjuvants.(6) However, in that study, there were three sensitizations, which makes the experiment relatively long for an acute model of asthma. The mechanism by which subcutaneous sensitization is effective without the use of adjuvants still needs to be clearly characterized. One hypothesis is that the subcutaneous tissue compartment has a larger and more effective number of antigen-presenting cells, which results in a more robust response to a given allergen, no adjuvants being required.
The cell counts in BAL fluid in the present study showed that eosinophil counts were lower in the groups undergoing subcutaneous sensitization with ovalbumin (21 days) and two intranasal ovalbumin challenges (40 µg) than in the group undergoing three ovalbumin challenges (Figure 2). This finding can be explained by the fact that fewer intranasal ovalbumin challenges induce less inflammation in the animal. However, in the context of the primary objectives of our study, this finding does not seem to be of great relevance.
One limitation of the present study was the fact that we did not measure cytokines or ovalbumin-specific IgE or conduct pulmonary function testing. Because we did not investigate any disease mechanism or any mechanism of antigen sensitization in the present study, we believe that the analysis performed, with a major focus on eosinophil activity in the lungs (eosinophil levels in BAL fluid, eosinophil levels in histological sections, and EPO levels in lung tissue), does not significantly change the final interpretation of the results in general. However, we understand that further studies involving a better characterization of the inflammatory response in the lungs are needed in order to validate our model. One question that remains unanswered is whether this short period is sufficient for a significant production of ovalbumin-specific IgE.
In conclusion, new models of asthma using lower doses of ovalbumin, a smaller number of sensitizations, a smaller number of intranasal challenges, and no adjuvants have shown an inflammatory response in the lungs, with a predominance of eosinophils (key effector cells in Th2 immune response). Our proposed model, of shorter duration, can be a future option for pre-clinical testing and the study of therapeutic targets in asthma, reducing the duration and cost of studies, provided that new studies are carried out and the appropriateness of the model is confirmed.
1. Taussig LM, Landau LI, editors. Pediatric respiratory medicine. St. Louis: Mosby; 1999. [ Links ]
2. Bates JH, Rincon M, Irvin CG. Animal models of asthma. Am J Physiol Lung Cell Mol Physiol. 2009;297(3):L401-10. PMid:19561139 PMCid:2739768. http://dx.doi.org/10.1152/ajplung.00027.2009 [ Links ]
3. Zosky GR, Sly PD. Animal models of asthma. Clin Exp Allergy. 2007;37(7):973-88. http://dx.doi.org/10.1111/j.1365-2222.2007.02740.x [ Links ]
4. Szelenyi I. Animal models of bronchial asthma. Inflamm Res. 2000;49(12):639-54. PMid:11211914. http://dx.doi.org/10.1007/s000110050642 [ Links ]
5. Eisenbarth SC. Use and limitations of alum-based models of allergy. Clin Exp Allergy. 2008;38(10):1572-5. http://dx.doi.org/10.1111/j.1365-2222.2008.03069.x [ Links ]
6. Conrad ML, Yildirim AO, Sonar SS, Kiliç A, Sudowe S, Lunow M, et al. Comparison of adjuvant and adjuvant-free murine experimental asthma models. Clin Exp Allergy. 2009;39(8):1246-54. http://dx.doi.org/10.1111/j.1365-2222.2009.03260.x [ Links ]
7. Renz H, Smith HR, Henson JE, Ray BS, Irvin CG, Gelfand EW. Aerosolized antigen exposure without adjuvant causes increased IgE production and increased airway responsiveness in the mouse. J Allergy Clin Immunol. 1992;89(6):1127-38. http://dx.doi.org/10.1016/0091-6749(92)90296-E [ Links ]
8. Hahn C, Erb KJ. The preclinical testing strategy for the development of novel chemical entities for the treatment of asthma. Curr Drug Targets. 2008;9(6):443-51. PMid:18537583. http://dx.doi.org/10.2174/138945008784533552 [ Links ]
9. Strath M, Warren DJ, Sanderson CJ. Detection of eosinophils using an eosinophil peroxidase assay. Its use as an assay for eosinophil differentiation factors. J Immunol Methods. 1985;83(2):209-15. http://dx.doi.org/10.1016/0022-1759(85)90242-X [ Links ]
10. Johnson JR, Wiley RE, Fattouh R, Swirski FK, Gajewska BU, Coyle AJ, et al. Continuous exposure to house dust mite elicits chronic airway inflammation and structural remodeling. Am J Respir Crit Care Med. 2004;169(3):378-85. PMid:14597485. http://dx.doi.org/10.1164/rccm.200308-1094OC [ Links ]
11. Fattouh R, Al-Garawi A, Fattouh M, Arias K, Walker TD, Goncharova S, et al. Eosinophils are dispensable for allergic remodeling and immunity in a model of house dust mite-induced airway disease. Am J Respir Crit Care Med. 2011;183(2):179-88. PMid:20732990. http://dx.doi.org/10.1164/rccm.200905-0736OC [ Links ]
12. Cates EC, Fattouh R, Wattie J, Inman MD, Goncharova S, Coyle AJ, et al. Intranasal exposure of mice to house dust mite elicits allergic airway inflammation via a GM-CSF-mediated mechanism. J Immunol. 2004;173(10):6384-92. PMid:15528378. [ Links ]
13. Brewer JM, Conacher M, Hunter CA, Mohrs M, Brombacher F, Alexander J. Aluminium hydroxide adjuvant initiates strong antigen-specific Th2 responses in the absence of IL-4- or IL-13-mediated signaling. J Immunol. 1999;163(12):6448-54. PMid:10586035. [ Links ]
14. Shapiro SD. Animal models of asthma: Pro: Allergic avoidance of animal (model[s]) is not an option. Am J Respir Crit Care Med. 2006;174(11):1171-3. PMid:17110653. http://dx.doi.org/10.1164/rccm.2609001 [ Links ]
15. Kool M, Soullié T, van Nimwegen M, Willart MA, Muskens F, Jung S, et al. Alum adjuvant boosts adaptive immunity by inducing uric acid and activating inflammatory dendritic cells. J Exp Med. 2008;205(4):869-82. PMid:18362170 PMCid:2292225. http://dx.doi.org/10.1084/jem.20071087 [ Links ]
16. Shi Y, Evans JE, Rock KL. Molecular identification of a danger signal that alerts the immune system to dying cells. Nature. 2003;425(6957):516-21. PMid:14520412. http://dx.doi.org/10.1038/nature01991 [ Links ]
17. Clausen SK, Bergqvist M, Poulsen LK, Poulsen OM, Nielsen GD. Development of sensitisation or tolerance following repeated OVA inhalation in BALB/cJ mice. Dose-dependency and modulation by the Al(OH)3 adjuvant. Toxicology. 2003;184(1):51-68. http://dx.doi.org/10.1016/S0300-483X(02)00583-8 [ Links ]
18. Van Hove CL, Maes T, Joos GF, Tournoy KG. Prolonged inhaled allergen exposure can induce persistent tolerance. Am J Respir Cell Mol Biol. 2007;36(5):573-84. PMid:17218615. http://dx.doi.org/10.1165/rcmb.2006-0385OC [ Links ]
19. Keller AC, Mucida D, Gomes E, Faquim-Mauro E, Faria AM, Rodriguez D, et al. Hierarchical suppression of asthma-like responses by mucosal tolerance. J Allergy Clin Immunol. 2006;117(2):283-90. PMid:16461128. http://dx.doi.org/10.1016/j.jaci.2005.10.019 [ Links ]
20. Holt PG, Reid M, Britten D, Sedgwick J, Bazin H. Suppression of IgE responses by passive antigen inhalation: dissociation of local (mucosal) and systemic immunity. Cell Immunol. 1987;104(2):434-9. [ Links ]
Correspondence to: Submitted: 26 March 2012. * Study carried out at the Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil.
Andrea M Rodrigues.
Centro Infant, Avenida Ipiranga, 6681, Prédio 60 (HSL), 2º andar, Parthenon
CEP 90619-900, Porto Alegre, RS, Brasil
Tel. 55 51 3320-3000, extension 2313
or 55 51 3320-3353
E-mail: firstname.lastname@example.com
Accepted, after review: 3 August 2012.
Financial support: Andrea Mendonça Rodrigues, Lucien Peroni Gualdi, and Raquel Giacomelli Cao are recipients of doctoral scholarships from the Brazilian Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, National Council for Scientific and Technological Development). Nailê Karine Nuñez is the recipient of a master's scholarship from CNPq.
Correspondence to:
Submitted: 26 March 2012.
* Study carried out at the Pontifícia Universidade Católica do Rio Grande do Sul - PUCRS, Pontifical Catholic University of Rio Grande do Sul - Porto Alegre, Brazil.


text:
May 2, 2004
LOS ALAMITOS, Calif. - The UCLA Women's Water Polo team closed out its 2004 season with a Third Place finish at the MPSF Championships after defeating Long Beach State 6-4. The Bruins finish the year with a 22-5 overall record.
UCLA got on the scoreboard first as Kacy Kunkel found the back of the net at 4:25 in the first quarter. Long Beach State tied the game at 1-1,but in the second period goals by Katie Flanagan and Molly Cahill would give UCLA a 3-1 lead. Another LBSU goal would make it 3-2 at half. In the third period, UCLA jumped ahead 5-2 with goals by Cahill and Kristina Kunkel. Long beach State would cut the lead to 5-3, but another Kunkel goal would give UCLA a 6-3 edge and ultimately the 6-4 victory.
UCLA goalkeeper Emily Feher continued her outstanding play at the MPSF Tournament, as she finished the game with seven saves.
UCLA vs. Long Beach State - 5/2/04
Teams 1 2 3 4 F
Long Beach State 1 1 0 2 4
Goals: UCLA - Molly Cahill 2, Kristina Kunkel 2, Katie Flanagan, Kacy Kunkel; LBSU - Gordin 2, Garcia, Stewart
Saves: UCLA - Emily Feher 7; Long Beach State - Hawkins 12

subset: commoncrawl


text:
The Workforce Center is hosting a series of job fairs to hire workers for General Dynamics IT to support a call center in what was once the Sallie Mae building in the Lynn Haven Industrial Park.
The company is looking to fill about 1,400 positions, and will emphasize filling supervisory and management positions at the upcoming job fairs. Recruiting for Customer Service Representative positions will also take place.
The first job fair will be July 30 from Noon until 5:00pm. Additional fairs will be held throughout the month of August job fairs will be held each Monday from 9:00am to 1:00pm and each Tuesday from Noon to 5:00pm.
To help facilitate the hiring process, Maria Goodwin, Director of Workforce Services for the Gulf Coast Workforce Board asks that job seekers first obtain a referral for the positions by registering in Florida's job matching system on the Employ Florida website (link below).
Once registered, job seekers interested in the GDIT positions should sign into their account, create a profile, search for the job numbers listed below, and follow the instructions in the job order. This can be done online, or in person at the Workforce Center, which offers free access to internet connected computers. Job seekers should bring their resumes to the job fair and wear appropriate interviewing attire, as GDIT will be conducting interviews on the spot.
The full set of open positions is listed in the link below.


text:
There's a new lottery app in town, at least for Illinois residents, and that very first real lottery app is already courting controversy, reports The Chicago Tribune. Indeed, this Internet Deals Examiner downloaded the free app from the iTunes App Store onto my iPad, simply to get a feel for the news-making app, and to spy out what's sure to be a coming trend across plenty of states nationwide.
"Anything's possible," touts the Illinois lottery app's opening page, proclaiming users "must be 18+ and within the State of Illinois to play. Know your limits and play responsibly."
That last admonition is exactly what gambling addiction experts are worried about: that lottery apps like these provide too easy of a gateway for people with a propensity to gamble to play irresponsibly. To that end, the app has set a minimum purchase of $5 – and an upper limit restriction that doesn't allow anyone to gamble away more than $150 daily or $1,050 weekly or $3,500 per month, totals that could still be a hefty sum for those who can't afford such numbers.
Few downloads, modest sales
Thus far, the official app has only experienced approximately 128,000 downloads and less than $1 million in ticket sales, quite low considering the convenience that players in a huge city like Chicago could undergo from popping onto their iPhones, iPads or Android devices to play the latest Powerball, Mega Millions or Lotto Extra Shot games.
Alas, app-creator Northstar hasn't put a lot of funding into advertising the app yet, although the company hopes to attract younger people who are more used to making purchases via the convenience of their mobile devices.
Most lottery tickets still sold via retailers
Business Week reports that nearly 100% of lottery tickets are still sold at the places we're used to seeing them: gas stations, convenience stores and grocery stores, just the way those retailers like it, in order to get a share of the winnings, if so allowed via their individual regulations.
Outside of the addition of individual states' apps is the addition of the LotteryHUB app, which allows for users to add various states' lottery information to their settings as well. The app is designed by Shoutz, out of Austin, Texas. Therefore, a writer like me can scan instant scratch-off tickets via the official Ohio lottery app to check for winners, and also find additional information via the LotteryHUB app.
With the ease of gaining money through things like an online cash advance, lottery tickets purchased via mobile apps and a plethora of other income-gaining procedures that used to require that consumers leave their homes to retrieve the money, this 21st century manner of making money requires quite a bit of self-discipline and wisdom to make proper monetary decisions. But for those of us who learn to play the high-tech game correctly and take advantage of the best of the digital cutting edge lottery landscape, we just may discover that we are among the first millionaires – made so without having to leave the comfort of our plush sofas.


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Bear Pond
LOCATION: Bear Pond, Waterford
Date: October 11
Time: 6:15 a.m.
Equipment: 40mm lens
"I'd gone by Bear Pond in the dark, so I hadn't noticed the gorgeous granite boulders, definitely one of my weaknesses! I have a hard time passing them by."
Please Note: This information was accurate at the time of publication. When planning a trip, please confirm details by directly contacting any company or establishment you intend to visit.


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Saturday February 8, 2014: Another quiet day, but one that offers an opportunity for creative expression, unique ideas and perhaps even the opportunity to access information you didn't know you had... so throw away logic and analysis and ride on the wings of your imagination.
Look carefully at where you find yourself "stopped" or "frustrated", then imagine the kind future you'd want to have. Don't limit yourself... Don't let self-doubt set in... Just unleash your imagination on your problems and come up with novel solutions.
You are in the midst of a personal process that offers extraordinary potential. Confidence and patience will be needed. You must not rush. It will take time to let new hope, vision and pragmatic ways of reorganizing yourself germinate within you.
Today's Quote
"If your life were a book and you were the author, how would you want your story to go? That's the question that changed my life forever."- Amy Purdy,Living Beyond Limits, TEDx talk
(Watch Amy's inspiring TEDx talk in the video above)
Please note: The sky speaks to you if you will let it. Every day it delivers a message that each person hears in a uniquely different way. What do these daily self help messages mean to you and your life? To find an astrologer or to find out more about Patricia ~ click here.


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Design Precision: Contemporary Remodel in Bronxville
From Margaret Everton on August 31, 2010 in New York
This remodel by Resolution: 4 Architecture makes me happyvery, very happy. The Rulon-Miller home in Bronxville, New York received an addition and total overhaul that ramped up the edginess of the existing structure, while also giving something of a peaceful ambience back to the surrounding environment.
Cedar planks on the homes exterior keep the rest of the structure from feeling stark. They also break up the contemporary design with varying colors and textures.
Materials are light, clean and high-quality, creating a clutter-free space, as seen in the bathroom.
Stark and somewhat eerie may be the best words to describe the bath, though this gives the homeowners a blank slate to work with.
Its not the most decked-out home in the world, but this Resolution: 4 Architecture remodel was perfect for homeowners that wanted to emulate the modern, contemporary style they admired. And they did so with excellent precision.


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Poor Martha
Sunday, July 22. Luke 10:38-42
Jul 04, 2001 by Garret Keizer
Then she was my high school sweetheart, now she is my wife of 25 years, but we still laugh about that evening when, sitting close on the couch in my living room, we were momentarily startled by a raucous noise directly overhead. Groaning and banging like a poltergeist, something seemed about to take the roof off the house, not to mention the glow off the evening. It turned out to be my mother, running a vacuum cleaner in the attic.
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Frida Gustavsson's modeling career took off in 2009. Then, the young model opened the Fall Valentino Haute Couture show in Paris and caught the eyes of top designers and fashion journalists. From that point on, her modeling career started to rise and during the Fall/Winter 2010-2011 season she walked on no less than 70 catwalks, among them Alexander McQueen, Carolina Herrera, Etro, Dsquared2, Missoni, Nina Ricci, Ralph Lauren, Chanel, Christian Dior, Fendi, Burberry Prorsum, Calvin Klein, Jean Paul Gaultier, John Galliano, Just Cavalli, Lanvin, Louis Vuitton and Marc by Marc Jacobs.
Though Gustavsson did not manage to repeat her achievement, her appearances on the catwalks were still mesmerizing. During Spring of 2012, she opened the Theirry Mugler show and closed the Carolina Herrera, McQueen and Prabal Gurung shows. During the Fall 2012 season, she worked it as the beautiful closure for the Armani Prive' fashion show in Paris.
Apart from fashion shows, Frida Gustavsson was photographed for many successful ad campaigns throughout the years. Prada, Anna Sui, Etro, Marc Jacobs, Max Mara, H&M, Just Cavalli and Spormax are just a few of them.
During those campaigns, Gustavsson posed for some of the world's best photographers: Patrick Demarchelier photographed her for the H&M Summer 2011 campaign, Mario Sorrenti was behind the camera on the Jill Stuart Summer 2012 campaign, and on during the Just Cavalli photoshoot, Gustavsson posed for photographers Mert Alas and Marcus Piggott.
The fact that Gustavsson is familiar with the fashion world, made her want to become more than just a pretty face. Her dream is to become a fashion stylist. Her main interests are creating patterns and product design.


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Parking enforcement officer is ready, willing, always a good citizen
Helping others drives Patricia Carver toward volunteerism
Patricia Carver, a parking enforcement officer at Shasta College and a member of Shasta College Chapter 381, is always ready to help.
"My community is so small," she said. "We do the best we can, but more often than not, we really need the help."
To that end, Carver volunteers for just about everything. Weeknights, she serves as a volunteer usher for the Redding Convention Center. If someone needs her assistance, she's ready to spring into action.
"For the most part people just need help and I'm here to do what I can," Carver said.
Carver's passion is helping. Working as a fundraising volunteer, she provides valuable assistance to the Shasta Women's Refuge. She enjoys knowing her help goes toward raising the money the organization needs to keep going.
"Without the two main events we put on each year, the Women's Refuge wouldn't be able to provide such important support to women hurt by domestic violence," she said.
In the fall, Carver works the annual Cornucopia, a business-oriented food and wine tasting and sampling event that includes a silent auction.
"I make sure everything with the silent auction goes smoothly," said Carver.
During the spring, Carver is always front and center to assist with the event. "Whatever I can do to help!" she said.
Carver also belongs to the Trails and Bikeways Council of Greater Redding. Last year, the group cleaned up several tons of trash.
"If we all just picked up after ourselves, everything would be a lot cleaner," Carver said. "It really is simple. Someone has to do it — I guess it's me."
That's a mantra for Carver, who also donates blood regularly.
"I was doing it every eight weeks, but the doctors told me I was getting a bit run down, so I spaced it out a little," she said.
Carver's passion for community involvement extends to CSEA as well. She has served as chapter president, job steward, regional representative and as a job steward mentor. She's even applied for the statewide job steward mentor program.
Carver's greatest hope is that as she gets older and is able to step back, newer and younger members will take the lead.
"It has been wonderful to see so many 'newbies' getting involved," she said. "I'll always be here to help, of course. I'll be here until I drop."
Carver finds it a little awkward to accept praise for her volunteerism.
"I just do things to help other people. Isn't that what being a good human being is all about?"
Still, her wheels are always turning as she constantly thinks about what else she can do to help others.
"Most things in life take a group effort," she said. "It's always better with a solid group behind you, just like negotiating with a district. If management knows you are all in sync, you can do just about anything."


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ABC interview: "International war may be declining, but according to journalist Peter Hartcher and Professor John Mueller, we should remain vigilant. While both agree that concerns about China and the Taiwan issue are certainly justified, if China's rise occurs without conflict, 'it will be the first time in history that a new power has risen to rival the reigning hegemony without a war.'"
Are wars on the decline? - ABC Late Night Live program claims it is
Current Status: Blessed (1)
Seeded on Wed Feb 8, 2012 5:27 AM
keyboard shortcuts: V vote up article J next comment K previous comment


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Cynthia Anne Mclemore Wood
Published: Wednesday, October 2, 2013 at 5:57 p.m.
Last Modified: Wednesday, October 2, 2013 at 5:57 p.m.
Cynthia Anne Mclemore Wood, 55, of Paul Musgrave Road, died Monday, Sept. 30, 2013, at the Kate B. Reynolds House in Winston-Salem.
A funeral will be at 2 p.m. Saturday at St Andrews Church, 310 Biesecker Road.
Reader comments posted to this article may be published in our print edition. All rights reserved. This copyrighted material may not be re-published without permission. Links are encouraged.


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Right off the bat, let me say:I am a big bag kind of girl.Even prebaby, I liked my handbags to be large and in charge and to come with plenty of room for all my stuff. (Plus, I just like the look of an oversized tote.)
So of course immediately Iwas drawn to the look of the Storksak Tania—its size is perfect for my style and taste, and I was taken with it from the get go by looks alone. But as I've learned before, sometimes good looks aren't enough when it comes to baby gear. You've got to have something that works for your lifestyle.
The outside of the bag is a fun metallic, a "pearlescent nylon with a waterproof finish."It's a great material for a diaper bag. It's durable and wipeable, and maintains its luster, making the bag's looks hold up for a long time (a super bonus when it comes to a baby carryall!) Another plus for me are the outside pockets. I'm forever toting around a water bottle (gotta stay hydrated when you're nursing!) and find it most convenient to be able to access my H2O by popping it in and out of an outer pocket of a bag instead of having to find a place for it inside. Ditto baby bottles! There are roomy pockets on the ends of the bag as well as larger pouch pockets on the front and back of the bag (with a handy inner pocket in one that's perfect for my cell phone).
Straps are another part of baby bags that take a beating, and this is another place that the Tania shines. The straps are strong and reinforced at the top, and though I've had the bag for only a fraction of the time that I'll be using it, I can already tell they won't show wear and tear easily.
Some people have a strong preference about how their diaper bags close and open—some like snaps, some like for the bag to remain open, some like zippers. I'm a zipper girl (a one snap closure seems wishy washy to me—either close all the way or stay open, Isay!) and that's exactly what the Storksak has. I like to be able to hide my plastic bag of wet clothes and crumpled receipts from the public.
Inside, you'll find a thermo-insulated bottle holder that keeps fluids warm or cool for up to four hours, plus a changing pad (for those on-the-go changes in places with questionable surfaces). In addition, there are pockets lining the sides and also a key ring, which Ifind immensely helpful. Lovers of big bags are notorious for losing their keys in the cavernous insides of their purse! From the minute I started using the bag, I would put my keys on the inner ring (it attaches via easy-snapping clip for removing and inserting) and Ithink it might have added 10 whole minutes back onto my week when you do the math.
Diaper bag choices are almost like name choices—so much of it comes down to personal preference. What works for one mom may not work for another. I absolutely adore this bag and think it was the very best choice for me. I think it will last through more than one baby for sure, making the price worth it, and it works with dressy outfits and casual outfits alike, making it my go-to bag for almost any day.
To buy:storksak.com


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Seated in the left center is the alchemist holding up a flask while looking at manuscripts that rest on a tapestry-covered table. A man holding a vessel, a woman, and another person watch the alchemist state the diagnosis. To the far right of a globe and vessels is a young assistant working the bellows. In the background is a group of men watching a dental procedure.
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Kool-Aid package, General Foods
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Photo of the Week: New Prosperity in Nicaragua
January 25, 2013
Fatima del Rosario Estrada takes a moment to play with her granddaughter.
Fatima del Rosario Estrada takes a moment to play with her granddaughter as she roasts a pile of rice husks in her backyard in order to produce compost. Fatima makes the compost for the Tomatoya-Chagüite Grande cooperative in northern Nicaragua. She can roast as many as 240 bags in a month, earning $1 per bag. The extra income helps her buy clothing, shoes and better food for her family.
"We used to eat just beans and cheese," Fatima says. "Now we also have pasta, rice, meat, chicken and vegetables."
With TechnoServe's assistance, vegetable farmers in the communities of Tomatoya and Chagüite Grande have turned their cooperative into a successful business that is creating new prosperity for families. Learn more about our work with the cooperative.
Related Blog Posts
Cashing in on Chips in Kenya
Hannah Karanja seized an opportunity to add value to potato products; now she's working with TechnoServe to expand her business.
Following in Her Father's Footsteps
How TechnoServe supported a long-time family business in the transition of ownership from father to daughter
Three Unorthodox Lessons for Latin America's Entrepreneurs
TechnoServe's global entrepreneurship director challenges conventional wisdom about how to help businesses grow.


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Thomas Mencl, 61, died Thursday at the Mercy Regional Health Center in Manhattan.
He was born on Aug. 5, 1952 in Chicago, Ill.
Memorial services for Mr. Mencl will be announced at a later date by the Yorgensen-Meloan-Londeen Funeral Home, 1616 Poyntz Avenue, Manhattan, Kansas 66502.
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It is truly amazing of the high caliber of talent pouring out of the Chicago area churches. Julian Key Thomas attends Providence church on Ashland Avenue and has since the age of eight. Providence is where Julian learned to play piano and sing. Currently, Julian is a singer and actor. His latest song is It starts with a dream. Julian's new movie is Escapement, a Jae Williams film.
If you ever attended a Bulls Basketball game then you have seen the Bulls Kidz performing before a tough crowd of basketball lovers, this is where Julian began performing before thousands on any given night at the age of 10. Julian is dashing around the United States drumming up press and interest for his music. LUTG RADIO aired an interview with Julian Keyz earlier today. Julian is preparing for the Escapement movie premiere at the Paramount in Boston, MA. The Chicago native is hoping for big love from the Chicago market when the movie makes its way to Chicago.
Julian expects to do a concert this summer in Chicago. Expecting more details from the Chicago Native Julian Keyz.
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One of the biggest reasons for my purchase of RIM was because of Apple. When RIM created the smartphone market it was what it was. Email, Text and Chat, 100 million people. Apple expanded the market 10 fold to over a billion people with its invention and for that I am grateful.
RIM is now re-entering a market that is 10x larger than when it began trading on the exchange. Even if the percentage of ownership of users is 10% (which I believe it may eclipse 15% or 20%) in the next 2 years that means their user base can double.
You could bring 80 million people to 160 million in no time. That is 80 million handsets plus the kicker of carrier fees. Revenue's would eclipse $30 billion. This is definitely a long bet but it is speculative. Do you have the cahones?
Read my posts for 3 years they are clear. It was short RIMM. I have no ended that position for a bit now and have purchased at 6.29,6.36,7.24,8.24,8.53 and 9.05.
This has a long way to go. 1 to 2% moves will add up fast with 10% pops. Don't want too many of these day, just nice slow creep up and then a pop.
If this goes up too quickly I personally would just dump it and run. Something that has value will climb slowly over time (months). Something that is manipulated and artifically driven suggests there is little value and lots of hype. So be aware.


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Another photo of Blakely Field
Barbara was born in the house in the lower center. It has since been torn down. "I didn't tear it down"
Photo Comments
Thank you Lewis. The Blakely name is one of Welch's most famous. They did a lot for the town. I was small, but I do remember Mayor Blakely. Thank you for checking out my site. I will add some more pictures soon. I have been lazy. :-)
Jul. 21, 2012 By:
Thank you for sharing the wonderful memories of Welch. My late father, William T. (Tug) Blakely, who played baseball at Welch HS and received a baseball scholarship to WVU, would be proud - as would my late uncle, John Blakely, former mayor of Welch and for whom Blakely Field was named.
Jul. 19, 2012 By:
Lewis Howard Blakely II
Mary Ellen I remember you. You had a brother Tommy. So good to hear from you. You were a cute little very blond girl.
Mar. 1, 2010 By:
I was 6 yrs old when we left Blakely Field. Oh this picture brings back memories. God Bless who ever submitted them. Mary Ellen Bishop Gilmore
Feb. 23, 2010 By:
Mary Ellen Bishop Gilmore
You can see why we got a lot of "home run" balls in our yard.
Nov. 3, 2005 By:
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From Wikiquote
Listening is the act of hearing something and paying attention to what is heard.
- But yet she listen'd—'tis enough—
Who listens once will listen twice;
Her heart, be sure, is not of ice,
And one refusal no rebuff.
- Lord Byron, Mazeppa (1819), Stanza 6.
- The young watch television twenty-four hours a day, they don't read and they rarely listen. This incessant bombardment of images has developed a hypertrophied eye condition that's turning them into a race of mutants.
- Federico Fellini, in I'm a Born Liar: A Fellini Lexicon (2003), "Younger Generation".
- Understanding does not occur when we try to intercept what someone wants to say to us by claiming we already know it.
- Hans-Georg Gadamer, "Aesthetics and Hermeneutics" (1964), in Philosophical Hermeneutics, translated and edited by David E. Linge (University of California Press: 1976), p. 102.
- When people talk listen completely. Most people never listen.
- Ernest Hemingway, letter of advice to a young writer, reported in Malcolm Cowley, "Mister Papa", LIFE magazine (January 10, 1949), Volume 26, No. 2, p. 90. A longer version appears in Hemingway's Across the River and Into the Trees (1967): "When people talk listen completely. Don't be thinking what you're going to say. Most people never listen. Nor do they observe. You should be able to go into a room and when you come out know everything that you saw there and not only that. If that room gave you any feeling you should know exactly what it was that gave you that feeling".
- The one who recites the Qur'an and the one who listens to it have an equal share in the reward.
- Muhammad Mustadrakul Wasa'il, Volume 1, Page 293.
Hoyt's New Cyclopedia Of Practical Quotations[edit]
- Quotes reported in Hoyt's New Cyclopedia Of Practical Quotations (1922), p. 461.
- He holds him with his glittering eye—
* * * * * *
And listens like a three years' child.
- Samuel Taylor Coleridge, The Ancient Mariner, Part I, Stanza 4. Last line claimed by Wordsworth. See note to his We are Seven.
- Listen, every one
That listen may, unto a tale
That's merrier than the nightingale.
- Henry Wadsworth Longfellow, Tales of a Wayside Inn (1863-1874), Part III. The Sicilian's Tale. Interlude Before the Monk of Casal-Maggiore.
- In listening mood she seemed to stand,
The guardian Naiad of the strand.
- Walter Scott, The Lady of the Lake, Canto I, Stanza 17.
- And this cuff was but to knock at your ear, and beseech listening.
- William Shakespeare, The Taming of the Shrew (c. 1593-94), Act IV, scene 1, line 66.


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STRACHE firstname.lastname@example.com_current.manitoulin_expositor 2004-06-23 published
Audrey STRACHE
In loving memory of Audrey STRACHE who passed away at the North Bay General Hospital in the presence of her family, on Tuesday, June 15, 2004 at the age of 71 years.
Cherished mother of Chris and Sylvia, both of Burlington, Audrey is also survived by their father Peter STRACHE. Dearly loved sister of Sheila COWPER of North Bay and Marjorie WILKIN of Little Current. Predeceased by her parents the late Walter and Jean GLANVILLE.
Special aunt and dear friend of Nancy PIDGEN. Fondly remembered by several nieces and nephews and her beloved pug Oscar. Formerly of Burlington, Audrey worked at Appleby Place prior to retiring. Friends called at the Island Funeral Home, Little Current, on Saturday, June 19 from 12: 30 pm until the memorial service in the chapel at 2 pm. Interment of ashes in the Mountain View Cemetery. Arrangement entrusted to the Martyn Funeral Home.
S... Names ST... Names STR... Names Welcome Home
STRACHE - All Categories in OGSPI
STRAIN firstname.lastname@example.com_current.manitoulin_expositor 2004-02-11 published
-In loving memory of a dear brother and uncle, Harold, who passed away March 1, 1980.
He meant so much to us
But nothing we can say
can tell the sadness in our hearts
As we think of him each day.
He always was true and tender
He lived his life for those he loved
And those he loved, remember.
Sadly missed by Gary, Elva, David, Jim and Karen
S... Names ST... Names STR... Names Welcome Home
STRAIN email@example.com_current.manitoulin_expositor 2004-04-07 published
Beatrice Lila (SCOTT) BEANGE
In loving memory of Beatrice Lila (SCOTT) BEANGE (1916-2004.)
On Saturday, April 3rd, 2004 Beatrice entered the presence of her heavenly Father for all eternity.
"We are confident, yes, well pleased rather to be absent from the body and to be present with the Lord." 2 Cor. 5: 8
Beatrice was the eldest of seven children born to Sarah (STRAIN) and William SCOTT of Mills Twp. After graduating from Normal School in North Bay, Beatrice spent several years teaching at area schools: Perivale, Burpee, Silver Water and Cockburn Island. In 1944 she married Charles (1917-1993) of Ice Lake and took up residence in Toronto as a homemaker. After raising sons Norman and Scott she returned to teaching in East York for a number of years. The purchase of a cottage on Lake Kagawong in 1978 enabled Beatrice to spend many pleasant summers back on the Manitoulin Island with family and Friends. The year 1996 saw Beatrice celebrating her 80th birthday and moving into Sheperd Terrace in Toronto.
Beatrice is survived by sons Norman, and wife Pauline of Toronto, and Scott, and wife Joanne of Thunder Bay, grandchildren Daniel, Amy, Michael, Charles, Julia, Natelie, and James and siblings Margie BLACKBURN, Maria McDERMID and John and Fred SCOTT. Friends and relatives may call at the Culgin Funeral Home in Gore Bay from 10: 00 - 11: 00 am on Saturday, April 10, 2004. The funeral service will be conducted at 11: 00 am with Reverend Ray KLOETSTRA officiating. Interment will follow at Mills Cemetery.
S... Names ST... Names STR... Names Welcome Home
STRAIN email@example.com_current.manitoulin_expositor 2004-09-01 published
William Matthew MATHESON
In loving memory of William Matthew MATHESON, February 3, 1924 - August 26, 2004, a resident of Silver Water, died at the Mindemoya Hospital on Thursday, August 26, 2004 at the age of 80 years.
He was born at Elizabeth Bay, son of the late Norman and Bertha (MORDEN) MATHESON. Matt had worked for the Department of Transportation, as a mechanic, from 1947 until his retirement in 1980. He was very active in his community, having been founding Fire Chief of the Robinson Fire Department, had served on many Boards of the local Service Board for Robinson Township, and had been a very active member of the Snowdusters, grooming countless miles of trails. Matt had a variety of interests other than his community involvement. He enjoyed snowmobiling, boating, flying, travelling and loved horses and music. He also enjoyed visiting with people. He will be sadly missed by his loving and loved family and numerous Friends. Beloved husband of Ollie (TENNISWOOD) MATHESON. Matt and Ollie would have celebrated 59 years of marriage next month. Precious Dad to Lyla (Peter) MUNT of Rocky Mountain House, Alberta, Norman (friend Linda) and Don (Virginia) of Silver Water. Dear Grandfather of Stephen, Judy, Robert and Katerina, Greg and Jessica. Great-grandfather of David, Ashley, Kathryn, John, Brittany and Logan. Dear brother of Vera FALLS, Anne HOPKINS, Audrey HARPER and Rhea WATT. Predeceased by brother Charles. Brother in law of Myrtle MATHESON, Pat (Ken) RICKARD, Hazel (Henry) REUBLE, Margaret (Ross) STRAIN and Don (Venus) TENNISWOOD. Also survived by many nieces and nephews. The funeral service was conducted in the Wm. G Turner Chapel on Saturday, August 28, 2004 at 11 am with Pastor Eriwn THOMPSON/THOMSON/TOMPSON/TOMSON officiating. Interment in Silver Lake Cemetery, Silver Water.
S... Names ST... Names STR... Names Welcome Home
STRAIN firstname.lastname@example.com_current.manitoulin_expositor 2004-11-17 published
--In loving memory of a dear father and grandfather, David, who passed away November 22, 1982.
Your smile has gone forever and your hand we cannot touch.
We have so many memories of you, Dad, we loved so much.
Sadly missed by Garry, Elva, David, Jim and Karen.
S... Names ST... Names STR... Names Welcome Home
STRAIN firstname.lastname@example.com_current.manitoulin_expositor 2004-12-22 published
Ruby Mae GREENMAN
In loving memory of Ruby Mae GREENMAN, April 18, 1912 - December 13, 2004.
Ruby GREENMAN, a resident of the Wikwemikong Nursing Home and formerly of Barrie Island, died at the Nursing Home on Monday, December 13, 2004 at the age of 92 years. She was born in Gordon Township, daughter of the late Wesley and Emily (LOWERY/LOWREY/LOWRIE/LOWRY) STRAIN. Ruby had been a resident of Barrie Island since her marriage to Thomas James GREENMAN in 1935, until moving to the Nursing Home about 5 years ago. In her earlier years, she had been a member of the Women's Institute. Ruby had a variety of interests which included knitting, quilting and watching her soap operas. Ruby will be remembered as a loving mother and grandmother who enjoyed cooking for her family and guests, and deeply loved her home and family. Ruby was predeceased by her husband Thomas in 1982. Loving mother of Robert, Doug and Rod all of Barrie Island. Predeceased by son Ernie. Proud grandmother of John, Janis, JoeAnn, Jeanna and Julie and 14 great grandchildren and three great great grandchildren. Predeceased by brothers Wes and Fred and sisters Freda, Viola, Audrey and Ethel. Also survived by many nieces and nephews.
Friends called at the Culgin Funeral Home after 7 pm on Wednesday. The funeral service was conducted in the Wm. G. Turner Chapel on Thursday, December 16, 2004 at 11 am with Pastor Maxine McVEY officiating. Spring interment in Gordon Cemetery. Arrangements in care of Culgin Funeral Home.
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Eleanor Mary CLARK
In loving memory of Eleanor Mary CLARK, September 28, 1915 to January 11, 2004.
Eleanor CLARK, a resident of Kagawong, died at the Manitoulin Health Centre, Mindemoya, on Sunday, January 11, 2004 at the age of 88 years.
She was born in Scarborough, daughter of the late Thomas and Margaret PATTERSON. Eleanor married Donald CLARK in 1940, and moved to Kagawong where they raised their six children.
Eleanor was very active in the United Church of Canada, teaching Sunday School and as leader of the Explorer Group and was thrilled to recently be presented with a lifetime membership in the United Church Women. For many years, Eleanor drove the local school bus and later enjoyed owning and operating her own General Store in Kagawong. She spent many hours knitting, sewing, gardening and was an avid reader. Eleanor was predeceased by her husband Donald MARSHALL CLARK in March of 1990. Loving and loved mother of Allen and his wife Dianne of Callander, Margaret and husband Tom STRINGER of Green Bay, Marilyn and husband Mike FARQUHAR of Little Current, Ken and friend Shelley of Callander, Ann and husband Eric PARSONS of Capreol, and Jim and Melody of Kagawong. Proud grandmother of Greg CLARK (Michelle,) Colleen VANDERWEERDEN (Richard,) Sharon CAMPBELL (Gaetan,) Leanne STRINGER, Dwayne STRINGER (predeceased,) Heather HANSEN (Robert,) Janyn STRINGER (Tim,) Jennifer FARQUHAR (fiancé Thanos,) Aaron FARQUHAR (Kristin), Evan FARQUHAR, Tim CLARK (predeceased), Nancy CLARK, Curtis KOSKI (Jennifer), Kristie KOSKI, Allison PARSONS (Paul), Sean PARSONS (Melissa), Curtis CLARK and Amy CLARK and will be sadly missed by her 11 great grandchildren. Dear sister of Margaret ALLEN of Newcastle and Len and wife Mona PATTERSON of Toronto. Friends called the Culgin Funeral Home on Monday, January
12, 2004. The funeral service was conducted in the William G. Turner Chapel on Tuesday, January 13, 2004 with Reverend Mary Jo ECKERT TRACY officiating. Spring interment in Kagawong Cedars Cemetery.
Culgin Funeral Home
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North Hendren Cheese Cooperative
Black River BlueA classic American blue from the North Hendren Cheese Cooperative which has been making cheese for 80 years. The recipe for this blue is well-guarded, and for good reason! The texture is slightly softer than butter- a nice creamy backdrop for the bright blue flavor. Read More
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Fourme d'AmbertThis cheese is in rare Fourme. Time in our caves yields a sweet, super-creamy bite that will convert even the staunchest blue cheese hater. Read More


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LIVE Signing Day Webcast On Tap At TexasTech.com
Texas Tech Athletics will host a live (free) Signing Day Webcast.
Jan. 27, 2012
National Signing Day Webcast (8 a.m. - 11 a.m. CT) | Tuberville News Conference (2:15 p.m. CT)
Texas Tech Athletics will provide unprecedented coverage of the 2012 National Signing Day (#TTUSigningDay) on Wednesday, Feb. 1, through its world-wide home located at texastech.com.
For the first time, Texas Tech will produce a free three-hour webcast dedicated to National Signing Day that will begin at 8 a.m. (CT) and conclude at 11 a.m. (CT) on RaiderVision All-Access (texastech.com). The show will provide instant analysis of the 2012 football signing class as soon as each NLI is received and verified by university officials. A live camera will give viewers the opportunity to see inside the "War Room" where coaches and administrators gather to process the NLI's as they come off the fax machine.
The webcast will also feature interviews with the following; assistant football coaches, Tommy Tuberville, athletics director Kirby Hocutt, senior quarterback Seth Doege, senior defensive back Terrance Bullitt and mid-year signees who have enrolled for the spring semester. The show will also include segments with Texas Tech Head Soccer Coach Tom Stone and Texas Tech Head Track & Field Coach Wes Kittley as they unveil their respective signing classes.
The show will be hosted by Robert Giovannetti, John Harris, Sonny Cumbie and Courtney Davis.
In addition to the signing day webcast, fans can follow breaking news updates through Twitter (@techathletics/@blaynebeal - using the hashtag: TTUSigningDay), Facebook (facebook.com/ttuathletics) or through text messaging (text RAIDERS to 51234 to sign up). Fans can also following a live blog and signing day updates on Signing Day Central located at texastech.com.
Live Video Events
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Miami Marlins: SS Jose Reyes tabbed with 500,000th MLB error
Jose Reyes after Saturday's game was surprised to learn from his agent he committed the 500,000th error in major league history.
So who determined Reyes was the culprit? The folks at baseball-reference.com. The website has a seasons index through which they track all-time stats since 1876. It came to the staff's attention MLB was nearing the 500,000th mark. Even baseball-reference.com admits this is not 100 percent accurate. It's probably pretty close.
Reyes as is usually the case was good-natured about the whole thing, saying: "I don't to want to make history that way. When [my agent] told me, I said, 'Really?' It's OK. It is what it is. They track everything."
The Marlins through Saturday had totaled 2,217 errors, including 96 this season.


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Feb. 15, 2012
DAVIS, Calif. - The No. 8 Stanford softball team falls at UC Davis, Wednesday, as the Aggies posted a 2-1 win at La Rue Field in Davis, Calif. The Cardinal drops to 4-2 overall, while UC Davis moves to 2-4.
Junior pitcher Teagan Gerhart earned the start in the circle, throwing 3.0 innings with three strikeouts. Freshman Nyree White entered the game in relief in the fourth and took the loss, dropping to 0-1.
Stanford escaped the first inning unscathed as UC Davis put two runners in scoring position with no outs. After a lead off double by right fielder Cassandra Ginnis, centerfielder Megan Guzman reached on an error by Stanford third baseman Hanna Winter. Fortunately, Gerhart was able to retire the next three batters without any damage done to the scoreboard.
Both teams were held scoreless in the second. The Aggies would strike first in the game in the bottom of the third on an RBI double by designated player Rachel Miller.
Stanford tied the score in the top of the seventh. With runners on first and second, a wild pitch by Thweatt allowed junior Jenna Rich to score from second and sophomore Tegan Schmidt to advance to third. The Aggies would get the final out of the inning and have one last chance for the win.
With one out in the bottom of the seventh, UC Davis catcher JJ Wagoner hit a triple to deep right field. Back at the top of the order, Ginnis ripped a single up the middle to score Wagoner and seal the win.
The Cardinal hosts the Stanford Nike Invitational this Friday-Sunday, Feb. 17-19 at Smith Family Stadium. Stanford takes on Pacific in its home opener on Friday at 2:30 p.m. The Cardinal will also face Bradley and Nevada over the weekend.


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marie25 wrote:Oh yes. I already have my preorder in for it. gamestop has it for preorder.
This message was edited 1 time. Last update was at 08/05/09 04:46 AM
badtothebone322 wrote:Are you buying the first EP, World Adventures?
Writin_Reg wrote:You can count on it. This is exactly what I put in the suggestion/new ideas section on the Sims 2 boards when they had the Sims 3 section there. I even named Egypt. France, and China or Japan and to make it an adventure. I called my idea Around the Workld Adventures. I also suggested an African Safari but instead of hunting animals to shoot it was for photography purposes and you would hunt for rare jungle plants for trophies. The African Safari is the only thing missing from my idea from the sounds of this Adventure EP. I will totally be looking forward to it - for sure. I already told the hubby to put it on his calender. hehehe
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text: The Most Prestigious Law School Admissions Discussion Board In The World. \ The most prestigious law school discussion board in the world. CLICK TO VIEW SCHOOL-RELATED + OFF-TOPIC THREADS (Expert Mode) Search: Most recent threads · Most active threads created last 24 hours / this week / this month Israel is a Jewish state. 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It is the Office of Sustainability's vision to showcase a living laboratory where students study and participate in the development of social equity, economic stability, and environmental stewardship. At the Flagstaff campus, students live, learn, eat, play, make purchases, use energy, and participate in every possible aspect of life—NAU is a micro-community! As such, NAU has the opportunity to demonstrate to larger cities, the nation, and the world what a truly healthy and sustainable environment can look like.
If you have an idea for how to help NAU become more sustainable, contact us to see if you can turn your idea into an internship!
Resources for Office of Sustainability Employees


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TUCSON, Ariz. ?C University of Arizona menˇ?s tennis head coach Tad Berkowitz announced today the signing of five star recruit Jason Zafiros to a national letter-of -intent.
Zafiros, a 5-foot-10 player from Bradenton, Fla., is ranked as the No. 35 prospect overall and the No. 8 player in the Southeast by tennisrecruiting.net.? Zafiros, who preps at the Nick Bolletieri Tennis Academy, has an overall record of 31-14 and is 5-2 vs. other five star recruits.
ˇ?Weˇ?re very excited to have Jason join our team next year,ˇ± said Berkowitz. ˇ?He should be able to come in and compete right away.ˇ±
Arizona opens its spring season on Jan. 25 at home against Gonzaga.


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Ben Mitchell
Mechanical Engineering '06
Ben Mitchell grew up in a devout household in the central Upper Peninsula of Michigan where emphasis was placed on helping others.
He traveled with his father to places such as Kenya and Peru on short-term medical missions. When he was in Kenya, a missionary explained that engineers have the greatest potential to influence the largest number of people. During high school, Ben participated in a youth mission trip to Tijuana. A result of the mission trips, his family decided to adopt three Vietnamese children when he was a junior.
All of these experiences had a great influence on Ben's outlook on life. He decided to major in mechanical engineering and enrolled at Lake Superior State University. Near the end of his senior year, he decided to volunteer for the Peace Corps to help others.
His two-year commitment began in the fall of 2007 and took Ben to Burkina Faso in western Africa. He resides in the village of Tansila, population 3,000, approximately 10 km from the Mali border. His host organization is the local cotton growers union. It works cooperatively with other villages to improve and expand their production techniques. He serves as an agribusiness advisor as a part of the Small Enterprise Development program which helps create market linkages and assists with the advancement of business skills.
Psychology Driven...
"LSSU is a relatively small school and I believe for this reason I have received a better education. The close atmosphere as opposed to a larger university has provided me with an opportunity to become acquainted with my professors. The professors therefore have an opportunity to assist with problems fairly easily, especially in upper level classes. The classes are fairly small also which provides one on one conversations which is very helpful."
[ more ]


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December 2, 1998
STANFORD, Calif. -- Flanker Troy Walters was named the Most Valuable Player of the 1998 Stanford Football Team and defensive tackle Willie Howard won the prestigious Al Masters Award at the annual Stanford Football Awards Banquet held December 1 at the Hotel Sofitel in Redwood Shores.
Walters, who caught 52 passes for 880 yards (16.9 ypc) and five touchdowns, was limited this season due to an ankle injury and played in just six full games. He set a Stanford record this season for career receiving yards and moved up to third on the school's all-time receptions list. Walters will return in 1999 for his senior season.
Howard, a junior from Mountain View, was presented with the Al Masters Award as the "player displaying the highest degree of leadership and respect of teammates." He also won the Russell-Huston Award as the "player exemplifying aggressiveness, exceptional performance and unherahlded efforts."
Howard and senior Donnie Spragan were named team captians for the 1998 season. Spragan, an outside linebacker who was recently named Second-Team All-Pac-10, also won the Jim Reynolds Award for the "senior player whose courage on the field and devotion to the game are an insipiration to all."
1998 Stanford Football Team Awards
Irving S. Zeimer Award (Most Valuable Player) - Troy Walters
Al Masters Award (highest degree of leadership and respect of teammates)-Willie Howard
The Vardell Award (excellence in both academics and athletics) - Jon Eide
Menlo-Atherton Trophy Award (outstanding freshman) - Joe Borchard
Deswarte-Eller Award (outstanding sophomore) - DeRonnie Pitts
Gundelach Award (outstanding junior) - Todd Husak
Jim Reynolds (courage on the field and devotion to the game are an insipiration to all) - Donnie Spragan
Greg Piers Award (outstanding service team players) - Ryan Wells (offense), Adam Scholze (defense)
Phil Moffat Award (outstanding special teams players) - DeRonnie Pitts (offense), Emory Brock (defense)
Russell-Huston Award (aggressiveness, exceptional performance and unheralded efforts) - Willie Howard
Frank Rehm Awards (outstanding back and lineman in Big Game) - Sharcus Steen, Austin Lee
1998 Team Captains - Willie Howard, Donnie Spragan


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Curtis LaMont Conway
Position: WR
Height: 6-1 Weight: 196 lbs.
Born: January 13, 1971 in Los Angeles, CA
High School: Hawthorne
College: USC (school history) (Conway college stats)
Drafted by the Chicago Bears in the 1st round (7th overall) of the 1993 NFL Draft.
Weighted Career AV (100-95-...): 61 (892nd overall since 1950)
Other SR Links: College Football at Sports-Reference.com
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The research for this page was done by Scott Kacsmar.
Read his extensive three-part article (Part I - Part II -
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View complete leaderboards for game-winning drives and comebacks.


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Twenty-four soldiers who bravely fought in World War II, Korea and Vietnam are to be awarded the Medal of Honor by President Obama next month, according to ABC News on March 11. The soldiers, who previously had been awarded the Distinguished Service Cross, had been overlooked for the Medal of Honor because of their ethnic and religious backgrounds. The Medal of Honor is the highest medal that can be awarded to a soldier in the four branches of military service, including the Army, Navy, Marines and Air Force.
President Obama stated that the Medal of Honor should have been awarded to these soldiers forty-four years ago:
"I apologize for the oversight. This should have happened 44 years ago. I was overwhelmed."
The Distinguished Service Cross, which is the second highest award that can be awarded a soldier, was an honor for the soldiers. However, Mitchell Libman, who advocated for Leonard Kravitz for over 50 years, felt that Kravitz and the other soldiers deserved to have their awards upgraded to the Medal of Honor. Libman never gave up and now the twenty-four soldiers are to awarded the Medal of Honor next month. Three of the soldiers will be awarded the Medal of Honor in person, while the other twenty-one soldiers are to receive their awards posthumously.
The soldiers fought in the following wars respectively: seven in World War II, eight in the Korean War, and nine in the Vietnam War. Libman was able to start the process that ultimately resulted in the awarding of the Medal of Honor to the twenty-four soldiers when he contacted Florida Congressman Bob Wexler. Wexler was able to get legislation passed in 2001 that ordered a review of the cases of Jewish and Hispanic soldiers. The review finally ended with the planned awarding of the Medal of Honor to the soldiers at the White House next month.


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Use into in a sentence
- They always put your other boots into inaccessible places. [5]
- But you say you reasoned yourself into it. [7]
- She did not yet stir, but there came into her face a slight inflection of confusion or perplexity. [11]
- Faithful and sincere, yet loving power, curious and adventurous, she must needs, without intention, without purpose, stray into perilous paths. [11]
- Only five hundred yards below the bridge was the second cataract, and she would never have waked if she had been carried into it. [11]
- Presently, however, the yacht slid out from the infolding land into an open sea that stretched before them to a silver-lined horizon. [9]
- She broke off, wringing her hands and staring into space, the ghost of the beautiful figure that had left the Opera House with shining eyes. [11]
- An elderly woman would not have sprung so nimbly into the skiff that was to convey her to the land. [10]
- Beyond that I would not go, into speculation I dared not venture. [9]
- I said I would never endure that horror again; that I would never again look into the grave of any one dear to me. [5]
- For better or worse John Hodder had flung his treasured beliefs into the crucible, and one by one he watched them crumble and consume away. [9]
- Laying down her work, she passed into the front hall. [11]
- I put into words what Charlotte Bronte put into actions. [14]
- With these cheerful words of encouragement he slipped out of a side door into what was evidently the court room, for Honora heard a droning. [9]
- Pierre, without a word, began cleaning his rifle, while Macavoy smoked, and sat looking into the distance, surveying the sweet warmth and light. [11]
This page helps answer: how do I use the word into in a sentence? How do you use into in a sentence? Can you give me a sentence for the word into? It may also be related to educator, lessons, schools, teach school learn, montessori school, and homework help.
Example sentences with the word into, a sentence example for into, and into in sample sentence.


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Tax ProcrastinatorsSubmitted By Liz Cooper, LCooperK2TV.com
It's getting to crunch time for you to file your taxes.
The deadline this year is Tuesday April 17th.
For all of you that haven't filed your taxes, you have less than a week.
Tax season can be a burdern for some people if they have endless amounts of forms to fill out.
But tax filers can choose to do it themselves or if planned properly, have an account do it for them.
You must fill out an extension form by April 17th if you don't have the proper paperwork ready to do your taxes.
The extension allows you six more months to file your taxes.


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« Star In Your Own Glamorous Fairy Tale | Main | Bright Young Thing »
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