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The present disclosure is related to the use of peptides for the purposes of vaccination, specifically it relates to peptides that induce a immune response in mucosa and more specifically it is related to a peptide included in a intranasal vaccine composition to elicit a protective immune response in mammals against illness caused by enterotoxigenic Escherichia coli.
At present it is estimated that over 1,500 million cases of diarrhea occurred every year worldwide, of which, 3 million ended in death. Of the total number of cases of diarrhea, 210 million are caused by the bacterium enterotoxigenic bacterium Escherichia coli, hereinafter designated as ETEC, of these episodes 380,000 cases ended in death (World Health Organization (WHO) State of the art of new vaccines Research & Development Initiative for Vaccine Research; Geneva, April 2003). Although the diarrhea caused by this microorganism occurs in groups of all ages, the mortality is more common in children under 5 years old, particularly when this illness occurs concomitantly with malnutrition, thereby the developing countries are specially affected. This bacterium also is the principal causal agent of the so-called traveler's diarrhea.
ETEC infection is acquired orally, principally through contaminated drinks or food; the bacterium overcomes the acidic conditions of the stomach until it reaches the small intestine, where, due to its Colonization Factor Antigens known as CFA's, it adheres to the intestinal mucosa and liberates its enterotoxins which are principally two, heat-labile enterotoxin known as LT and heat-stable enterotoxin or ST which are the factors responsible for the diarrhea (Gaastra W, Svennerholm A M. Colonization factors of human enterotoxigenic Escherichia coli (ETEC). Trends in Microbiology. 1996; 4(11): 444-452).
More than 20 different CFA's have been described in the literature, and of these three have been identified as the more prevalent, and they are known as CFA/I, CFA/II and CFA/IV that were reported by several authors, since there was detected between 50 to 75% of ETEC strains isolated from patients suffering from diarrhea all over the world, including Mexico, the majority are grouped in the family CFA/I. (Gaastra W, Svennerholm A M. Colonization factors of human enterotoxigenic Escherichia coli (ETEC). Trends in Microbiology. 1996; 4(11): 444-452. López-Vidal Y, Calva J J, Trujillo A, Ponce de León A, Ramos A, Svennerholm A-M, Ruiz-Palacios G M. Enterotoxins and adhesins of enterotoxigenic Escherichia coli are they a risk factor for acute diarrhea in the community? J Infect Dis 1990; 162:442-447, and Steinsland H, Valentiner-Branth P, Gjessing H K, Aaby P, Molbak K, Sommerfelt H. Protection from natural infections with enterotoxigenic Escherichia coli: Longitudinal study. The Lancet. 2003; 362). These types of CFA's induce protective immunity against the bacterium and they are pointed out as the most important among the strategies for the development of effective vaccines. (Middlebrook J L, Dorland R B. Bacterial Toxins: Cellular mechanisms of Action. Microbiological Reviews. 1984; 48(3): 199-221. McConell M M, Hibberd M L, Penny M E, Scotland S M, Cheasty T, Rowe B. Surveys of human enterotoxigenic Escherichia coli from three different geographical areas for possible colonization factors. Epidemiol. Infect. 1991; 106: 477-484).
Due to the epidemiological importance of ETEC, many of the efforts have been directed to the prevention of the illness by obtaining an effective and safe vaccine, (Levine M M, Kaper J B, Black R E, Clements A M. New Knowledge on Pathogenesis of Bacterial Enteric Infections as Applied to Vaccine Development. Microbiological Reviews. 1983; 47(4):510-550) however, to the date these efforts have been unsuccessful and a product that satisfies these needs has not been placed in the market yet. (World Health Organization [WHO] State of the art of new vaccines Research & Development Initiative for Vaccine Research; Geneva, April 2003).
One of the vaccines that is under development and that has passed to the phase of trials in human healthy volunteers from different geographical regions, was prepared in the University of Götemburg in Sweden, and it is based on subunit B of cholera toxin combined with 5 formalin inactivated strains of ETEC, which all together express the CFA's of most epidemiological importance on a global scale. (Quadri F, Ahmed T, Ahmed F, Sack B, Sack A, Svennerholm A M. Safety and Immunogenicity of an oral, inactivated enterotoxigenic Escherichia coli plus cholera Toxin B subunit vaccine in Bangladeshi children 18-36 months of age. Vaccine 2003; 21:2394-2403).
Other strategies for the development of vaccines have focused on the use of live bacteria, as it is seen in the work realized in the Center for Development of Vaccines CVD of the University of Maryland, using Shigella as vector for the expression of colonization factors and enterotoxins from ETEC. (Barry E M, Altboum Z, Losonsky G, Levine M M. Immune responses elicited against multiple enterotoxigenic Escherichia coli fimbriae and mutant LT expressed in attenuated Shigella vaccine strains. Vaccine 2003; 21:333-340).
Recently a vaccine has been prepared with a new technology of administration; by means of a patch for transdermal immunization containing the surface component of E. coli known a CS6 and the heat-labile toxin known as LT; which has been already tested in human volunteers, where an immune response of Th1 and Th2 type characterized by IgG2a and IgG1 respectively, was determined (Wenneras C, Firdausi Q, Prodeep K B, Bladley S and Svennerholm A-M. Intestinal Immune Responses in patients Infected with Enterotoxigenic Escherichia coli and in vaccinees. Infect. Immun. 1999; 66:3311-3316. Quadri F, Ahmed T, Ahmed F, Sack B, Sack A, Svennerholm A M. Safety and Immunogenicity of an oral, inactivated enterotoxigenic Escherichia coli plus cholera Toxin B subunit vaccine in Bangladeshi children 18-36 months of age. Vaccine. 2003; 21: 2394-2403. Byrd, W., and F. J. Cassels. 2003. Mucosal immunization of BALB/c mice using enterotoxigenic Escherichia coli colonization factors CFA/I and CS6 administered with and without a mutant heat-labile enterotoxin. Vaccine 21: 1884-1893. Helander A, Wenneras C, Quadri F, Svennerholm A M. Antibody Responses in Humans against Coli Surface Antigen 6 of Enterotoxigenic Escherichia coli. Infection and Immunity. 1998; 66(9):4507-4510). However, the immune response against the colonization factors was not consistent as shown by the prevalence of the illness in different geographical regions.
There is other type of developed strategies that including vaccines formed by colonization factors encapsulated in microspheres, or by means of the expression of the subunit B of LT in plants of tobacco, potatoes, tomatoes and bananas, but these strategies are highly expensive and nevertheless the conferred protection is low.
Another option can be a vaccine prepared with the common linear epitope of immuno-dominant sequences from CFA's which offered a broader spectrum and increases the level of protection from the ETEC infection (López-Vidal Y, Epitopos continuos y comunes presentes en las fimbrias de Escherichia coli enterotoxigénica (ETEC). Gac. Med. Mex. 1997; 133 (6): 511-525. Domínguez M, et al. Colonization Factor Antigenic I Peptide as Intranasal Vaccine Approach against enterotoxigenic Escherichia coli infection in hamsters. 5th National Symposium, Basic Aspects of vaccines, 1999).
Generally we can recount that different types of vaccines have been evaluated against diarrhea caused by ETEC and although many of these vaccines have demonstrated protective effectiveness against ETEC infection, some of them show certain side effects; another important problem to be considered in the design of vaccines against ETEC, is the variability in the prevalence of the different colonization factors, which prompted us to design a vaccine that is effective and safe against the different ETEC serotypes.
Intranasal administration can be an additional alternative for a vaccine against ETEC, which would add additional advantages, providing superiority over other vaccines. In this respect a great number of studies has been performed that demonstrated that the mucosa-associated lymphoid tissue known as MALT, is a common system, that is, that the stimulation in a certain site of the mucosa also known as inductor site elicits a response at local level, remotely or in the effector site, this represented a great advantage in vaccine design, with the form of transmucosal administration being greatly facilitated (Cripps A W, Kyd J M, Foxwell A R, Vaccines and Mucosal immunization. Vaccine 2001; 19:2513-2515). The use of the intranasal route as route of administration has as principal advantages the presence of a highly vascularized greater surface, the elimination of the use of syringes with the risk that this poses, the decrease of administered doses compared to the oral route which involves a decrease of adverse effects, easy administration, application to a great number of persons in relatively short times and the induction of antibodies and cells of the immune response (Zuercher A W. Upper Respiratory Tract Immunity. Viral Immunology 2003; 3; 279-289). There are studies that demonstrate that the intranasal administration can stimulate the production of secretory IgA at intestinal level, which provides a great advantage in the development of vaccines directed to the protection against intestinal pathogens (Hong-Yin Wu, Russell M W. Induction of mucosal and systemic immune responses by intranasal immunization using recombinant cholera toxin B subunit as an adjuvant. Vaccine 1998; 16(2/3):286-292).
Vaccine strategies to improve immunity at the level of the mucosa include the use of these alternate routes together with adjuvants. (Colonization Factor Antigenic I Peptide as Intranasal Vaccine Approach against enterotoxigenic Escherichia coli infection in hamsters. Domínguez M, et al. 5th National Symposium, Basic Aspects of vaccines, 1999). Recently, nasal mucosa has been used as inductive site that has showed to increase the immune response at level of the mucosa both locally and distally such as in intestinal mucosa, respiratory mucosa and genital mucosa. Administering certain antigen to the mucosa associated to lymphoid tissue is a way of eliciting immune response in sites distal from mucosa. (Byrd, W, and F. J. Cassels. 2003. Mucosal immunization of BALB/c mice using enterotoxigenic Escherichia coli colonization factors CFA/I and CS6 administered with and without a mutant heat-labile enterotoxin. Vaccine 21:1884-1893. Van Ginkel F W, Nguyen H H, McGhee J R, Vaccines for Mucosal Immunity to Combat Emerging Infectious Diseases. Emerging Infectious Diseases. 2000; 6(2):123131.)
For the above, is still of great relevance to count with an effective vaccine against ETEC infection that can be applied globally and through a route of administration that allows a rapid, easy application and that could minimize the adverse effects that could be generated.
The present disclosure relates to a peptide vaccine that can be administered intranasally, containing a common linear protein epitope from different ETEC colonization factors that can be recognized by sera of patients infected with said bacteria.
FIG. 1 shows a summary of post-immunization results obtained from mice with CLE, CTB or the vaccine and from a control group that later was infected by ETEC H 10407; the percent of ETEC excretion is shown where: − corresponds to negative; + corresponds to 25%; ++ corresponds to 50%; +++ corresponds to 75%, and ++++ corresponds to 100%, and in respect to the occurrence of diarrhea: 0=no diarrhea; 1=loss of consistency of feces but without wet tail; 2=wet perianal region and tail, and 3=tail, paws and low abdomen wetted and with an inactive appearance. In this figure it can be appreciated that the developed vaccine has the ability to decrease bacterial excretion in addition to the signs of diarrhea, which is evident on day 3 after the infection.
FIG. 2 shows the results corresponding to subpopulations of CD4+ and CD8+ T lymphocytes of cells obtained from the spleen and Peyer patches after immunization of mice with CLE, CTB or the vaccine and a control group that later was infected by ETEC H10407 where it can be seen generally that the highest percentage of CD4+ and CD8+ T lymphocytes was obtained by the group vaccinated both in spleen and in Peyer patches.
FIG. 3 shows a integral graph where the percentage of cells producing intracellular cytokines of lymphocytes specific to vaccine obtained from spleen, inguinal lymphatic ganglions, lymphatic cervical ganglions and Peyer patches belonging to mice immunized with CLE, CTB or the vaccine was analyze. From the graph it can be observed that cytokines IL-4 and IL-10 were mainly produced for animals vaccinated in all the organs of study, but a low production of INF-gamma was also observed.
FIG. 4 shows the evaluation of ETEC infection that expresses CFA/I in a golden Syrian hamster model after vaccine administration where it can be observed that in the vaccinated animals there a decrease both the percentage of sick hamsters and the duration of diarrhea when compared to the control group, it can also observed that there is a decrease of the time of ETEC excretion.
FIG. 5 shows the levels of IgG antibodies against CLE and CFA/I of ETEC from vaccinated hamsters and control, 11 days after the infection with ETEC CFA/I where it can be appreciated that a significant increase occurs in IgG antibodies titre against CLE in the animals after the vaccination.
FIG. 6 shows antibody titres against CLE, CFA/I, CFA/II, CTB and LT in vaccinated hamsters and control 8 days after infection with ETEC and it can be appreciated clearly that after the vaccination higher titres of antibodies against CLE, CFA/I, CFA/II, CTB and LT were generated.
For the development of this vaccine, first the immune response that elicited the linear common epitope known as CLE associated to mucosal adjuvant, cholera toxin B subunit known as CTB, in a murine model and in a Syrian golden hamster model was characterized. Starting from the entire fimbria CFA/I, a 20 amino acids peptide amino acids named as CLE was identified and studied, said peptide is located in the linear sequence of enterotoxigenic Escherichia coli CFA/I fimbria from amino acid 33 to amino acid 52 herein identified as SEQ ID NO: 1, which contains a terminal cysteine in position 53, included to form protein complexes such as dimers, trimers and pentamers, with the trimers being predominant at 37° C., such that it will have stability in its configuration and its ability to facilitate recognition by class II receptor from major histocompatibility complex and T lymphocyte receptor could be increased. This peptide is capable of eliciting immune protective response against the principal serotypes of ETEC.
Synthesis of CLE was performed by conventional methods. CTB was purchased from Merieux Laboratory, France.
In order to elucidate the sequence of peptide CLE, the characterization of common linear epitopes for different fimbrias of ETEC was carried out, by means of synthesis of continuous octapeptides spanning the entire sequence of the colonization factor antigen I designated as CFA/I; using the method of Geysen in which the synthesis of peptides was carried out in polyethylene sticks such that each of them corresponded to each of the wells of an ELISA plate. This arrangement enabled the study of recognition by antibodies against sera from children under five years old who were afflicted with ETEC infection, sera of adults from endemic and not endemic zones, hyperimmune sera that were prepared by immunization of rabbits with ETEC strains carrying different colonization factors and anti-CFA/1 monoclonal sera. The essay was performed similar to an ELISA. Most of the sera from children belonging to the convalescent stage recognized all the octapeptides, with the recognition being higher for three of said octapeptides, same that were recognized by serum from an adult belonging to an endemic area. Whereas the anti-CFA/1 monoclonal antibody did not recognized any peptide. The recognition of octapeptides was designated epitopes.
The identification of common and continuous epitopes in CFA/I by hyperimmune heterologous sera was very variable and it depended on each of the sera, and it was found that the colonization factors with higher similarity and that exhibited crossed antigenic response were CFA/I, the surface components of coli 1, 3 and 4 designated as CS1, CS3, CS4 and the putative colonization factor 0166 or PCF0166 and part of the sequence from common linear epitope CLE was recognized on these elements. CLE was selected because it was recognized by 100% of the children's sera after natural infection and by sera of adults from endemic areas for ETEC. From this finding, the peptide vaccine CLE of the colonization factors with B subunit B of cholera toxin CTB was developed in our laboratory, whose administration was the intranasal route due to the already mentioned advantages.
Circular Dichroism: A solution of 100 g/mL of CLE in trifluoroethane was used for the readings using a quartz cell of with 1 cm of optical pathway. Measurements were carried out in a JASCO J-715 spectrophotometer. The obtained spectrum is the average of five readings.
The spectrum of circular dichroism for CLE in PBS at pH 7.4 and room temperature 20° C., suggests the formation of a secondary structure. The spectrum indicates that CLE from CFA possesses a predominant structure of beta sheet that agrees with the prediction of a secondary structure performed in the Antheprot 5.0 software.
Dynamic light scattering: One milliliter of 1 mg/ml CLE diluted in 10 mm PBS, pH 7.4 was used to evaluate the dynamic light scattering at different temperatures of 5, 15, 25, 35 and 37° C. Hydrodynamic diameter of CLE was determined using the Zed ziser nano series (Mc Malvern, USA). It was found that CLE has the ability to form trimeric structures at physiological conditions of pH and temperature.
Having properly characterized CLE and evaluated its ability to elicit an protective immunological response, we proceed to develop a vaccine composition integrated with such a relation in its components, which enables to potentiate the protective effect of CLE, in this disclosure it is used preferably in a proportion of 8 parts of CLE to one part of a suitable adjuvant for use in mucosa, in this case the adjuvant CTB was used in a pharmaceutically acceptable aqueous vehicle for intranasal application in a pharmacologically effective dose, without the use of this adjuvant is limitative, simply it appears like selected in this form of the disclosure. The proportion used in this vaccine composition will vary according to the used adjuvant.
The following examples are presented to provide a better understanding of the subject matter disclosed herein and they are not intended to limit the disclosure to the same.
Peptide CLE was applied to pathogen-free, 6 to 8 weeks old, female BALB/c mice, which were randomly distributed into groups of five animals in each group. Peptide CLE, corresponding to SEQ ID NO: 1, was administered in a pharmacologically effective dose, that for this case was 50 μg/mL in a volume of 7.5 μL per mouse, intranasally, three times at 0, 7 and 28 days. The control group was provided with phosphate buffered solution PBS orally. The challenge was carried out one week after the last immunization, by administering by gastric route an inoculum of 6×108 CFU of strain H10407 of ETEC.
To evaluate the conferred protection, parameters such as the excretion of ETEC in feces by means of coproculture and identification by means of hemagglutination test in the presence of D-mannose were determined, (López-Vidal Y. Colonization Factor antigens of Enterotoxigenic Escherichia coli (Monoclonal Antibodies and Methods for Epidemiological Studies). 1990. Goteborg, Sweden. Doctoral Dissertation) and the determination of the presence of diarrhea signs was evaluated according to the severity with the following scale: 0 no diarrhea; 1 loss of the consistency of the feces but without wet tail; 2 region perianal and tail wetted and; 3 tail, paws and low abdomen wetted and with inactive appearance (Arredondo L J, Zaragoza S, Domínguez M, Willms K, López-Vidal Y, Cravioto A. Desarrollo de un nuevo Modelo Animal Experimental Hámster Sirio Dorado para el estudio de la infección por Escherichia coli enterotoxigénica (ETEC). Enf Infec Microbiol. 1997; 17(2):43-46).
We also characterized the immune response obtained for both studied groups for which cells from the spleen, lymphatic cervical ganglions, inguinal lymphatic ganglions and Peyer patches were obtained from mice on day 7 after the challenge with ETEC H10407. The adjusted cell suspensions obtained from each organ, were grown in the presence of antigens of CLE.
At the end of the cellular proliferation, the cells were labeled using antibodies to identify surface markers such as CD3, CD4 and CD8 and for the identification of intracellular cytokines such as IFN-gamma, IL-4 and IL-10. Once the cells were labeled, they were analyzed by flow cytometry.
The results showed that in relation to the excretion of ETEC, it was observed that on day 1, in both groups of animals positive coprocultures occurred at 100%, In the third day the positive coprocultures were of 75% for the control group and 50% in the group immunized with the peptide CLE. The presence of diarrhea in mice after the infection with ETEC was observed only with a change of consistency of the feces for the CLE-immunized and control groups, which continued to the fourth day in the latter group but from the fifth day, the consistency in feces was normal in all mice. These results can be appreciated in more detail in FIG. 1, which presents as a summary the follow up of mice after infection with ETEC H10407.
As for the evaluation of the immune response it was found that the response of T CD4+ and T CD8+ lymphocytes was slightly higher for the group immunized with CLE compared to the control group and it is necessary to emphasize that the increase of T CD4+ lymphocytes was about 2-fold in Peyer patches with regard to control group as it can be seen in FIG. 2.
As for cytokines response, a predominant production of IL-4 and IL-10 was observed in the group immunized with CLE, an increase being observed in the production of IL-4 of about 2.6 times in lymphatic cervical ganglions in the group immunized with CLE relative to the control and of 0.8 times in the production of IL-10 in the same organ, which is illustrated in FIG. 3.
Mice were immunized with CTB at a rate of 6 μg/mL by administering intranasally a volume of 7.5 μL and the control group was orally administered with a phosphate buffered solution, following the same methodology described in Example 1.
The obtained results are as follows: relative to the excretion of ETEC, it was observed that on day 1, both the control group and the group immunized with CTB exhibited positive coprocultures in 100%. To the third day the positive coprocultures were 75% for the control group and of 50% for the group immunized with CTB with a response similar to the obtained in mice immunized with CLE of the Example 1 being observed. Similarly to that observed for the mice immunized with CLE, those immunized with CTB exhibited inconsistency of feces up to the third day, but after the fourth day the consistency in the feces was normal in the CTB group and the control group as it can be observed in more detail in FIG. 1.
With this it was demonstrated that CTB has the ability to provide certain protection against infection caused by ETEC, but this protection it is not sufficient to counteract the signs of illness in mice such as the inconsistency of feces. As for the immune response it was observed that the subpopulation of T CD4+ increased 0.4 times in spleen in the group immunized with CTB relative to the control but T CD8+ subpopulation was very similar to the control group in the studied organs as can be observed in FIG. 2. The cytokines profile showed a preferential increase of IL-4 with a difference of 2.7 times being observed in lymphatic cervical ganglions and of 1.1 times in Peyer patches with regard to the control group. However, a considerable response of INF-gamma was obtained in inguinal lymphatic ganglions as it can be observed in FIG. 3. This demonstrates that the CTB, when it is administered alone to mice by intranasal route, induces an increase of immune response cells and cytokines of the predominant type Th1 such as IFN-gamma, thereby stimulating an immune cellular response, and it has already described that the protective response against ETEC infection is of humoral type.
Vaccine developed in the present disclosure containing the peptide CLE identified in SEQ ID NO: 1 was used in a 8:1 ratio of CLE:CTB administered intranasally in a pharmacologically effective dose, that in this case was adjusted to the weight of the experimental animals, with a final concentration of 50 μg/mL added with CTB at doses of 6 μg/mL, administering a volume of 7.5 μL following the same methodology of Examples 1 and 2.
The obtained results were the following: relative to the excretion of ETEC, it was observed that on day 1, both the control group and the group immunized with the vaccine exhibited positive coprocultures in 100%. At third day the positive coprocultures were 75% for the control group and of 25% for the group immunized with the vaccine. The presence of diarrhea in the mice after the infection with ETEC was observed only with a change of consistency of the feces for the control groups, and this event did not appear in the group immunized with the vaccine as can be observed in FIG. 1.
It was possible to determine that the vaccine developed in the present disclosure exhibits a synergistic effect between the peptide CLE and the adjuvant CTB for the protection conferred against diarrhea caused by ETEC. With regard to the control group, the group immunized with the vaccine presented an increase of T CD4+ lymphocytes of 0.5 times in the spleen and of 5.7 times in Peyer patches. This increase was observed also for the T CD8+ lymphocytes in the group immunized with the vaccine with 0.1 times higher in the spleen and 1.8 times higher in Peyer patches; when comparing to the control group as is observed in FIG. 2.
IL-4 production showed a considerable increase for the group immunized with the vaccine mainly in spleen and in inguinal lymphatic ganglions of 1 and 2.8 times higher, respectively, compared to the control group. The response in lymphatic cervical ganglions and in Peyer patches for the group immunized with the vaccine showed an increase of IL-4 production in comparison to the control group of 2.1 and 0.2 times higher respectively.
As for the response to IL-10, an increase of this cytokine was observed in all groups with regard to the control, although this increase was lower than that observed for IL-4. The group that showed a higher increase was that of Example 3 immunized with the vaccine compared to the control group which is of 19.5, 0.4, 8 and 6.8 times more in spleen, lymphatic cervical ganglions, inguinal lymphatic ganglions and Peyer patches, respectively. The highest percentage of IL-10-producing cells was obtained in lymphatic cervical ganglions being 8.1% higher in the group immunized with the vaccine.
Finally, although the percentages of producing cells of IFN-gamma were very poor in all the studied organs, an increase was observed in cervical and inguinal lymphatic ganglions with regard to the control group of 1.5 and 4.3 times respectively. Both in the spleen and in Peyer patches differences were not observed in the response of IFN-gamma as can be observed in FIG. 3.
These results indicate that the intranasal administration of the vaccine containing the peptide CLE added with CTB in a ratio of 8:1 respectively in a pharmaceutically acceptable vehicle has the ability to elicit a protective response of T lymphocytes T principally CD4+ in sites of immune response as the Peyer patches, which is of great utility because these are located in the small intestine which is the target site of ETEC infection. Also, the vaccine generated the production of cytokines such as IL-4 and IL-10 distinctive of a response of type Th2 that stimulates an immune response of humoral type that is the principal protective defense against illness caused by ETEC.
From the obtained results from Examples 1, 2 and 3 it can be concluded that the protection conferred by CLE when it is administered in mice by intranasal route increases when it is administered in a formulation that contains an adjuvant, which in this case is the adjuvant CTB so that there is an synergistic effect between these two components of the vaccine.
Female, 6 to 8 weeks old, Syrian golden Hamsters were intranasally immunized with peptide CLE corresponding to SEQ ID NO: 1 at a dose of 50 μg/mL added with CTB at a dose of 6 μg/mL by administering a volume of 7.5 μL of each component. Each hamster was immunized on day 0, 7 and 28. Blood samples by retrorbital route and samples of saliva were collected placing small strips of cellulose in the oral cavity, before the first immunization and 7 days after the last immunization. To the hamster control group was administered with a phosphate buffered solution.
To evaluate the conferred protection, parameters such as the excretion of ETEC in feces by means of coproculture and identification by means of hemagglutination test in the presence of D-mannose were determined as was already reported (López-Vidal Y. Colonization Factor antigens of Enterotoxigenic Escherichia coli (Monoclonal Antibodies and Methods for Epidemiological Studies). 1990. Goteborg, Sweden. Doctoral Dissertation) and the determination of the presence of signs of diarrhea which was evaluated in accordance to severity: 0, without diarrhea; 1, loss of the consistency of the feces but without wet tail; 2, region perianal and tail wetted and; 3, tail, paws and low abdomen wetted and with inactive appearance (Arredondo L J, Zaragoza S, Domínguez M, Willms K, López-Vidal Y, Cravioto A. Desarrollo de un nuevo Modelo Animal Experimental Hámster Sirio Dorado para el estudio de la infección por Escherichia coli enterotoxigénica (ETEC). Enf Infec Microbiol. 1997; 17(2):43-46).
To evaluate the immune protective response, the antibody titres were determined by ELISA against: CLE, fimbria CFA/I and against CTB and LT. And to determine the ability of these antibodies to agglutinate ETEC having different types of fimbria belonging to the CFA/I family an essay of agglutinative capacity was carried out with a suspension of formalin inactivated bacteria (1×108 CFU) expressing fimbria CFA/I and sera from hamsters immunized with the vaccine and infected with ETEC expressing CFA/I. Furthermore, the neutralizing activity of serum anti-CTB antibodies from hamsters immunized and challenged against cholera toxin (CT) and against LT (5 μg/ml) by cellular permeability in New Zealand rabbits, was determined.
The obtained results demonstrated regarding the excretion of ETEC and the presence of signs of diarrhea that the average of duration of diarrhea caused by ETEC was 3 times higher in the control hamsters than in the immunized hamsters. The average percentage of ETEC excretion increased 2.5 times in the control group as it shows in FIG. 4.
Regarding the antibody titre against CLE, CFA/I and enterotoxins by means of ELISA, the levels of IgG antibodies, in hamsters immunized with the vaccine and control hamsters were compared, after being infected by ETEC expressing CFA/I; the differences showed to be significant against CLE, but not against CFA/I which is shown in FIG. 5. On the other hand, high titres were detected against CLE, CFA/I, CTB from V. cholerae and LT from ETEC. These titres remained constant, before and after the oral infection, with ETEC expressing CFA/I as is shown in FIG. 6. Also, it was found that these anti-CLE antibodies were capable of agglutinating significantly to the whole bacterium, which expresses CFA/I at titre of 2.6 and the antibodies against CTB, at titre of 2; they were capable of neutralizing in 100% the activity of (5 μg/ml) the toxins LT and CT.
These results demonstrate the effectiveness of the use of the intranasal vaccine against diarrhea caused by ETEC since in the model of Syrian golden hamster high titres of antibodies were generated against CLE, CFA/I, LT from ETEC and CTB from V. cholerae, in addition that these antibodies were capable of inducing protection from the diarrhea caused by ETEC expressing CFA/I.
Finally, being ETEC an important causal agent of infantile diarrhea and of the traveler on a global scale, its identification is not performed routinely in the laboratory, partly due to the need to provide a rapid and opportune treatment, nevertheless, the peptide of the SEQ ID NO: 1 disclosed herein, is found as an immunodominant epitope in strains of ETEC belonging to the CFA/I family, for this reason it would be an excellent candidate for the development and making of a diagnosis reagent specific for this family of ETEC.


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Today, I am very proud to say that California took a historic step in the right direction. My Golden State voted to ban the possession and sale of shark fins.
This is exciting for many reasons, not least of which is the fact that our oceans' apex predators play a critical role in the health of the ecosystems that support and feed people across the globe.
But what I personally find so promising is the realization that we, as citizens, wield enormous power. When passionate and diverse stakeholders come together around a unified goal -- anything is possible. Sheer determination and grassroots outreach by organizations like NRDC, C.O.A.R.E. and Sea Stewards lifted the chorus of voices that wanted to put an end to the barbaric practice of shark finning.
Today we celebrate a victory for sharks and a victory for everyone who made phone calls, tweeted, shared a link, signed a letter or took a photo because sharks are definitely worth more alive than dead. (Special shout-out to folks like Leo DiCaprio and Ian Somerhalder who spread the word to their very large network of followers, and to Fred Buyle and William Winram for their stunning photos).
As my colleague Leila Monroe has often shared in her blog, a shark's value is squandered when sharks are killed for short-term gain from the sale of their fins. And because California is one of the largest markets for shark fins outside of Asia, by stopping the fin trade here we can help put an end to this type of destruction in international waters.
More than 1/3 of shark species are threatened with extinction as a result of the international shark fin trade, with some populations declined by 99%. Although 26-73 million sharks are killed every year, just for their fins, many states and nations around the world are taking bold action to stop the wasteful "finning" of sharks: Chile, one of the biggest fin exporters, recently banned shark finning in their waters, joining the Bahamas, Honduras, the Maldives and Palau. In the U.S., Hawaii passed a first-of-its-kind law banning the sale, possession, and distribution of shark fins in the state) last year, and similar measures have passed just this summer in Washington State and Oregon.
Thanks to the leadership of Assembly members Paul Fong (D-Cupertino) and Jared Huffman (D-San Rafael) -- the authors of the bill to ban shark finning (AB 376) -- and to the groundswell of public support that came pouring in, AB 376 will move onto to the governor's desk for a signature and then into the California books as a law.
As Paul Fong said "I'm pleased that California can take part in the worldwide movement to protect these important creatures, and that we can continue to provide leadership in important environmental matters."
Long live the sharks!

subset: commoncrawl


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Save money on your car insurance
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Originally published February 19th 2016
If you're involved in an accident and your car has to go in for repairs, you probably won't want to be forking out for taxis or relying on public transport while your motor is off the road.
But can you be sure your fully comprehensive policy will provide you with a courtesy car? Or does your policy require you to pay extra on top of your premium to ensure you get a replacement vehicle?
What is a courtesy car?
A courtesy car is a vehicle provided by a garage or repair shop for use while your car is being repaired. A courtesy car may be provided as standard on your car insurance policy, or you may need to purchase it as an optional extra.
Policy details
As with all things insurance, the provision of a courtesy vehicle varies from insurer to insurer, with it's important to check your policy to make sure you have exactly the cover you're after – and there are no nasty surprises lurking in the wings when you have an accident.
To highlight how courtesy car provision can vary between insurers, and why it's important to check the terms and conditions, we have a tale of two policies.
Mixed experience
The first is from Clare Francis, our former editor-in-chief at MoneySuperMarket, who was told the cover she got as standard on her Post Office policy would only provide her with a courtesy car "if the garage has got one in" – which means some motorists could be in for an unpleasant surprise if they're expecting a replacement vehicle.
Our second tale comes from a customer who, while insured with Direct Line, was given a car of comparable size to her own when it went in for repairs, as per the terms of its Guaranteed Hire Car Plus cover – Direct Line does not provide a courtesy car as standard.The car she was given while her Mercedes in for repairs was a top-of-the range Jaguar, which she thought was a little excessive for a hire car, but Direct Line's Guaranteed Hire Car Plus cover stipulates drivers will be given a comparable vehicle while theirs is being repaired.
If you don't think you'll need this, there's the cheaper Guaranteed Car Hire option, which entitles you to a small, 1000cc car while yours gets fixed. Always check the details to make sure you're not paying for extra cover you don't need or want.
Industry standard?
So, is there an industry standard for courtesy car provision, or is it down to the discretion of individual insurers? We asked the industry trade body, the Association of British Insurers (ABI), where it stands on the matter. It said:
- If you are involved in an accident and no other driver is involved, then whether or not you are entitled under your own motor policy to a courtesy car while yours is being repaired will be down the provisions of your policy.
- If you are involved in motor crash with another driver, but you are at fault, you will not be able to claim against the other driver for repair, or replacement of your vehicle. If you claim on your motor policy then the above applies.
- However, if involved in accident with another driver and they are at fault then you can claim against their insurer for courtesy car while yours is off the road. But your claim must be reasonable in that it must be for a similar type vehicle as yours.
In short then, it's more or less down to the insurer.
The only rule that applies across the board is that if you're at fault in a crash with another driver, you must claim on your own policy and can't claim off the other driver's policy – which stands to reason.
So just how much does courtesy car provision vary from insurer to insurer? We checked out some policy outlines from some of the UK's biggest insurers to find out.
What the insurers say – a market selection…
Aviva/Quote Me Happy
A courtesy car is standard with Quote Me Happy and is an additional cover option with its parent company, Aviva. With both brands, you can claim more than once over the term of the policy.
This means, under the terms of a standard Quote Me Happy policy, or under the additional cover option offered by Aviva, you will be given a courtesy car for the duration of the repairs being carried out, so long as these are being carried out by an approved repairer and you were not at fault for the accident.
Aviva's courtesy car option will give you a small, 1 litre hatchback for the duration of the repairs unless you pay for 'enhanced courtesy', in which case you'll get a 1.6 litre 5-door model.
Quote Me Happy's standard cover will give you a courtesy car if yours is damaged in an accident and you use its approved repair network.
If you choose the additional car hire option, you'll get hire car if car is stolen (insurers generally don't offer courtesy cars for stolen vehicles) or written off and can keep car for 21 days or until settlement cheque is sent.
Subject to availability, a free courtesy car will be provided to you for the duration of any repairs, provided your car is repaired by one of its approved repair centres.
A courtesy car is offered as part of standard fully comprehensive and third party fire & theft (TPFT) cover if car is fire damaged and repaired by an approved garage.
For an extra premium you can take out Guaranteed Car Hire and Guaranteed Car Hire Plus, both of which offer a courtesy car in the event of fire damage, accident damage, damage as a result of theft, if car is stolen or written off or if the car is awaiting repair. It will also provide a courtesy car if you use your own garage for any repairs.
Guaranteed Car Hire provides a car for a maximum of 14 consecutive days, Guaranteed Car Hire Plus provides a car for a maximum of 21 consecutive days.
Churchill fully comprehensive customers will receive a courtesy car as standard no matter how many claims are made or if there are multiple claims in one year.
Direct Line
No courtesy car as standard, but you can take out one of two optional extras for courtesy car provision. Guaranteed Hire Car: a hire car will be supplied for up to 14 consecutive days.
The vehicle will be a small 1000cc hatchback car or similar and is available to customers who have comprehensive policies.
Guaranteed Hire Car Plus: a hire car will be supplied for up to 21 consecutive days. The vehicle will be of a similar physical size to your own and this is available to customers who have comprehensive policies.
A free courtesy car is provided while your car is being repaired by one of its recommended repairers.
A courtesy car is not available if your car is stolen or if it is considered to be a total loss or if your policy is cancelled or lapses during the period of repair.
A courtesy car is standard and provided for each valid claim a customer makes. Esure will not limit the number of claims for a courtesy car while the car on cover is being repaired.
1st Central
A courtesy car will be provided if your vehicle is being repaired by one of the insurers' approved repairers.
A courtesy car will not be provided if your car has been stolen, is beyond economic repair or if you choose a repairer not on the insurer's approved panel.
The insurer is unable to guarantee a courtesy car for owners of cars originally produced for sale outside the European Union.
Providing you use Hastings's appointed approved repairer, a courtesy car will be provided for the duration of the repairs.
This is guaranteed and provided as standard for comprehensive cover.
The type of car provided is what is classed as a Class A courtesy car – which is a standardsmallhatchback (usually a 3-door, 1 litre vehicle such as a Nissan Micra or Ford KA). There are no limits on the number of claims you can make within a policy year or the number of times a courtesy car will be supplied.
Courtesy cars are an optional extra for LV= car insurance customers and cost £19.90 a year.
If a customer does choose to include a courtesy car on their policy, they won't be restricted to how many times the firm will provide a courtesy car.
For example, a customer can have multiple claims in a year and still receive a courtesy car on each occasion.
After loss or damage, a small hatchback under 1200cc will be provided if the repairs are carried out by its Selected Repairer Service.
This will be provided for up to 21 days or until 4 days after payment of the claim has been issued, whichever is soonest, if your car is a total loss and cannot be driven safely, or is stolen and not recovered.
More Th>n
If you have comprehensive cover, you'll receive a courtesy car as standard if your car needs repairs following an accident and you use a recommended repairer (excluding total loss).
All More Th>n customers who have fully comprehensive insurance get a courtesy car if they need it, and there's no restrictions on the amount of times it is provided.
It says that, "If a customer needs a courtesy car, they should have one."
Post Office
Use of a courtesy car is offered while yours is being repaired by one of its approved repairers following an accident covered by the policy.
As Clare Francis discovered, courtesy cars are subject to availability and the supplying garage's terms and conditions.
Sheila's Wheels
A free courtesy car will be provided while your car is being repaired by the firm's recommended repairers under a valid claim.All you pay for is the fuel.
A courtesy car is not available if your car is stolen or is considered to be a total loss. An additional car hire benefit means you will be given a courtesy car if your car is written off or stolen, up to a maximum of 21 days.
Fully comprehensive cover provides alternative car for maximum of 14 or 21 consecutive days in any one instance and a maximum of 21 or 28 days in any one policy year, depending upon the level of cover chosen.
This applies either while car is being repaired because of any damage following a claim or until your claim is settled if your own car is affected by fire, theft or is written off.
If a courtesy car can't be provided, the firm will repay your alternative travelling costs up to a maximum of £15 a day.
TPFT provides an alternative car until the claim is settled if your car is affected by fire or theft for maximum of 14 or 21 consecutive days in any one instance and a maximum of 21 or 28 days in any one policy year, depending upon the level of cover chosen.
Choose the right cover
As you can see, each insurer has its own policy on the provision of courtesy cars and most will not provide a replacement vehicle if yours is stolen or written off completely. So, as ever, the message is to shop around to make sure you get the exact cover you're after.
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By bringing together the most innovative companies in Asia and the West, the deployment of a variety of promising clean energy options can be accelerated. The climate challenge will be solved by multiplying opportunities for rapid development and deployment of low-carbon generating technologies, not by restricting engagement between companies in the world's most dynamic economies.
Investments by one country reduce the cost of that technology worldwide, increasing the likelihood that CCS will be widely deployed in time to help avert the worst consequences of climate change.
Advantages China Brings to CCS:
Working with China can speed CCS deployment and lower costs
Projects in China are completed in about half the time as ones in the West. This is not simply a function of permitting, but the entire procurement and construction schedule is faster.
Capital and labor costs in China are a fraction of their costs in the West. These lower costs are important not just in building large projects, but the play an important role in reducing costs associated with RD&D. The rising education levels in these countries combined with lower costs make them important potential centers of CCS innovation.
Investment funds, especially in China, are available for new energy projects and for innovative energy technology development. Some individual Chinese companies have energy technology R&D budgets as large as the US DOE R&D budget. Furthermore, CCS may be viewed as a potential export to the West which may drive CCS interest in India and China.
Because China has a thriving coal gasification and power development markets, most global power system and coal gasification expansion for the foreseeable future will occur in these markets. This growth can facilitate CCS.
Advantages the West Brings to CCS:
The West has deep experience with CCS technology and policies that support its growth
North America, the EU and Australia represent CCS resources (demonstration projects, experienced companies, etc.) that can support CCS technology development/deployment in China.
North America in particular has deep experience with EOR. In Texas alone, over a bilion tons have been injected for EOR since the 1970s. However in China, EOR-CCS experience is quite limited. But the potential to use EOR to advance CCS in China is high. Estimates from Advanced Resources International (ARI) for conventional CO2 EOR project that China has as much as 12 billion tons in potential CO2 demand (based on technical potential)- equivalent to the captured CO2 from 65GW of coal-ored power plants over a thirty year period.
Climate policies in the West, while limited in many respects, have driven the development of pioneer projects with high levels of integration. Experience with highly integrated CCS projects in lacking in the developing world. The West has extensive experience in EOR, a technology that is not widely practiced in the developing world. This know-how would accelerate CCS projects in Asia.
As a result partnerships could dramitically lower CO2 releases in both Asia and the West. The partnerships could:
Some examples of successful partnerships between the US and China are listed below:
CATF's China Project:


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Haven't made your Thanksgiving dinner plans yet? No worries. The Raddison New Rochelle is offering a great buffet — for a great price! For $36 per person ($21 for kids under the age of 10), you get a buffet full of soups (butternut squash or cauliflower), salad, and a string of traditional entrees. They'll have organic turkey with giblet gravy and baked pineapple ham, with sausage and mushroom stuffing. Then there's baked ziti, sweet potatoes, roasted garlic potatoes, haricot verts, creamed sauteed onions, and a whole lot more. And of course, there will be plenty of pie for dessert.
The Thanksgiving buffet will be served in the Empire Ballroom, near the hotel's lower lobby. Call for reservations: 914-576-3700.


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The name of head of the family was given as Sheik Yusuf Tanimola who was in his 30s.
Among Tanimola's four children were twins who were two years old.
Our correspondent gathered the residents of the area woke up on Tuesday to meet the family in a pool of blood.
Many of them claimed that they did not hear any strange sound and did not suspect any foul play until they saw the corpses of the family members.
It was gathered the house of the Tanimolas was a bevy of activities as residents and sympathisers thronged the residence.
The state Police Public Relations Officer, Abimbola Oyeyemi, said the command had deployed men in the community in a bid to unravel those behind the killing.
He said, "The Commissioner of Police has deployed men of the homicide section, the Federal Special Anti-Robbery Squad and Anti-kidnpping unit in the community.
"We are already profiling the identities of the killers; we will get them."


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Welcome to this website where you can find information about the horse's lymphatic system, conditions which affect it, and the use of manual lymph drainage and combined decongestive therapy with animals.
Unfortunately ill health has forced me to retire from practice since setting this site up but I hope it will still be of interest.
I started Equine MLD in 2006, after training to use manual lymph drainage with horses under the tutelage of Prof Berens von Rautenfeld, Hanover Medicine School. I had no idea until this finished that I was their first "overseas candidate" to qualify, through the Europäisches Seminar Für Equine Lymphdrainage. I returned to Britain full of enthusiasm for this treatment which could be used with so many equine disorders, including lymphoedema, a condition to which horses are prone but which, if veterinary manuals and horse magazines were to be believed, did not exist in the English language speaking world. In fact, it often seemed that they didn't possess lymphatic systems!
Although there are references to the lymphatic system going back nearly two and a half thousand years, historically it's been difficult to visualise, resulting in neglect when compared to other aspects of the body. It's largely through the pioneers of manual lymph drainage and combined decongestive therapy in the last hundred years that the science of modern lymphology was born, and its clinical practice developed.
Therefore my website, the first of its kind set up in English, was intended to educate about the equine lymphatic system as well as the use of manual lymph drainage. That there is a demand for such information has been confirmed many times by the range of enquiries I've received, and hearing from so many people has been an unanticipated pleasure. It's also been an education. Through working with horses in America and the United Kingdom, and through email conversations stretching literally all over the world, I have been able to gain understanding about the role of the lymphatic system in conditions affecting the horse which simply isn't available elsewhere, and to share this with horse carers.
I've also had the opportunity to undertake original research into the use of MLD to control Chronic Progressive Lymphoedema with Dr Verena Affolter, one of the team at UC Davis who discovered this disease, which has since been published in the Equine Veterinary Education Journal.
MLD is used by Olympic athletes for pre- and post competition maintenance which is therefore included here. Human MLD treatment was originally being developed for use with dogs before it was realised that horses are prone to lymphatic problems, and dogs and other animals are here too.
In the last two decades, advances in visualisation techniques have led to an enormous new interest in lymphatic research, confirming the background to manual lymph drainage and revealing that the lymphatic system has much greater importance and influence than previously realised. It's a truly exciting time, as much of this work suggests links to conditions affecting horses and hopefully this is starting to be recognised. Nevertheless, there's a lot of 'information' about equine lymphatic conditions, particularly on the internet but also in practice, which doesn't stand up to scientific scrutiny, and a lot still waiting to be discovered.
You may be looking at this site specifically because you're interested in finding out whether manual lymph drainage can help your horse, but I would invite you to also look at some of the more 'theoretical' content about the lymphatic system, and how new research may one day benefit horses too.
A word of caution. As lymphatic conditions become better known, interest in treating them by people without adequate knowledge to do so is growing. Do check a practitioner's qualifications or experience before letting them work on your animal.
You'll find sections on the equine and canine lymphatic systems and on conditions that specifically affect them, as well as conditions which benefit from MLD treatment. There's also a section on research into the use of MLD for laminitis and for Chronic Progressive Lymphoedema. There aren't many photographs of me doing MLD because it's a subtle manual technique and makes for uninformative pictures, but you will find some of my favourite horses, present and past.


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It was time to plan one last vacation before baby comes in July! Due to our busy spring schedule we decided to plan a winter getaway earlier in the pregnancy. We left cold and rainy Seattle behind and flew south to Los Angeles. We found a great deal on airfare using our favorite go-to trip planning site, Trip Advisor. Finding a hotel proved to be a little more difficult and we ended up spending quite a bit more then we normally do. We decided to throw caution to the wind and live it up one last time, booking ourselves into the Shore Hotel, located across the street from the ocean and steps away from the famous Santa Monica Pier. It is ranked the number one hotel in Santa Monica on Trip Advisor. We hoped it would be the perfect jumping off point for the oceanside adventure we hoped for!
Our flight landed at LAX just before 9:00 a.m. We caught a taxi and began the 8.5 mile journey to Santa Monica. Though the distance was short the length of the trip was not. We found ourselves amidst the morning traffic that Los Angeles is so famous for. We arrived at the hotel just after 10:00 a.m. and were staggered at the amount of cash we parted with as we paid the cabby for his services. Knowing it was far too early to check into our hotel we approached the front desk hoping to have them hold our suitcase while we did some exploring. The gentleman who helped us happened to be the manager and offered us a free upgrade to an ocean view room because the room we were supposed to have wasn't available yet. So, he let us check in hours early and gave us a free upgrade! Who could argue with that?!? The only snag was that our credit card put an immediate hold on when we attempted to use it…. We keep forgetting to alert our credit card company to our travels. After a lengthy stint on hold Bobby finally got the hold lifted and we checked in. Complimentary glasses of Champagne or Sparkling Cider are given at check in. As we entered our beautiful room we toasted each other and the long weekend ahead of us with cider!
After a quick respite in the room, we decided to go find some lunch. I kicked off my socks, slipped on my Toms, dug my sunglasses out and we headed out into the sunshine! Our hotel was very conveniently located just blocks from the charming 3rd Avenue Promenade shopping area so we headed there in search of sustenance. Chipotle was one of the first things we spotted and while it seemed kind of lame to eat at a chain restaurant, it also happens to be the food I have been craving most while pregnant, so it worked out perfectly! We sat in the small patio area and watched the small crowds go by. We were rather surprised by how few people were out and about on a sunny Friday afternoon.
After lunch we did some window shopping and then headed back towards the ocean. We explored the paths with beautiful plants situated above the beaches. We meandered along the paths for awhile before reaching a pedestrian bridge that crossed over the Pacific Coast Highway that barred our way to the beach. Once on the beach we strolled along the sand for a bit before heading to the paved pathway that was very popular with walkers, joggers, skateboarders, bicyclists, and rollerbladers.
We soon found ourselves exploring the small carnival area on the Santa Monica Pier. There were some tourists here, but again the crowds just weren't that bad, which we found rather surprising.
We hadn't been exploring long when fatigue set in for both of us. The day had started before dawn, in order to catch our early morning flight and we were ready for a nap! We headed back to the hotel for an afternoon siesta. Later in the day we ventured out to a grocery store to get snacks, and breakfast foods. We got an early supper and caught a bit of the sunset!
Our evening was spent trying very hard to stay awake to watch the opening ceremonies of the Olympics. We didn't make it through, I'm sad to say…
The following morning dawned a bit cloudy but still warmer and far more pleasant then the weather in Seattle, where we learned it was snowing! We decided to catch a bus to Hollywood. We did no planning for this trip whatsoever. Our list of must-sees was short. I wanted to see the hand and foot prints in front of the Chinese Theater, the stars on the Hollywood Walk of Fame, and Rodeo Drive. Bobby wanted to see a skate park that has been featured in several skateboarding videos. Every day Bobby would do reconnaissance and plan our day while I sat by, thankful to not have to do the planning and willing to go pretty much anywhere!
The bus ride to Hollywood was long, over an hour, but at $1.50 per person it was far more economical then a taxi! Thankfully, our stop to get off was conveniently located in front of a Jamba Juice so we stopped for a mid-morning snack before walking to the Chinese Theater. Seeing the handprints and the stars on the Hollywood Walk of Fame was really fun! The crowds were thick here and the hawkers trying to get you to join their bus tours were obnoxiously persistent.
Shopping was never in short supply and we soon found ourselves in yet another outdoor shopping area. While uninterested in the stores, we were interested in the view of the Hollywood sign we got at the top floor of the mall.
After exploring for a bit longer we decided to catch the bus and take it to Rodeo Drive were we decided we would find some lunch. We enjoyed window shopping along the glitzy and famous street. We felt like complete scrubs in our attire that would be considered perfectly acceptable in downtown Seattle. We were surrounded by some of the fanciest cars I have ever seen! Not to mention some of the most dolled up women I have ever seen!
We decided we would walk the 2.4 miles from our hotel to Venice Beach. There's a paved path the whole way there along the beach with lots to see and do in between! We first walked through Muscle Beach where Bobby showed off his muscles by playing on the outdoor gym equipment.
The boardwalk on Venice Beach is filled with an eclectic group of artists selling their wares at booths. After walking the length of the boardwalk we settled on buying some photographs from a local artist who was as colorful as some of his pictures! Apparently he was an extra in a movie a couple of decades ago, which he was incredibly proud of!
The skatepark Bobby had been hoping to see was there and was hoping with skaters young and old, talented, and not so talented. It definitely made Bobby ache for his skateboard that was busy gathering dust in rainy Seattle.
We walked along the pier and snacked on apples and graham crackers watching a group of surfers float in the rather still waters of the ocean. We weren't totally sure if the surfing is usually good here or if these guys just enjoyed really quiet waters…
We walked away from the ocean and towards the Venice Canal Historic District just a few blocks away. This charming and expensive neighborhood felt like a movie set in such a picture perfect setting. We saw two photo shoots as we explored the area.
We found lunch along Abbot Kinney Road, which according to GQ magazine, is the coolest block in America. Lots of high priced stores, and several appealing restaurants. We chose one that didn't have an enormous crowd outside waiting to be seated for Sunday brunch and were pleasantly surprised at the tastiness of the food! We walked along the road and back to the ocean path that would lead us back to our hotel. Along the way we found a section of the beach set up with dozens of slack lines and watched as some guys did some pretty sweet tricks! Bobby joined in on the fun!
We watched the sunset on the beach and celebrated reaching the 17th week in the pregnancy!
The final day of our trip ended up being one of our favorites. We met up with our friends, Kris and Ruth, who live in LA and we ventured out of the city and into the picturesque area of Topanga Canyon for a short hike.
Overall, this vacation couldn't have been more fun! We saw so many fun and unexpected things, spent some incredible quality time together, met up with old friends, and hiked in a beautiful desert setting. It was hard to leave the sunshine behind and return home to chilly, rainy Seattle. Latigo was happy to see us though!


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Your First Visit
How does therapy help?
Depending on your present situation and your reasons for seeking help, there are many benefits to therapy. If you are seeking diagnosis or treatment for a mental illness, therapy can help you better manage your symptoms, outbreaks, and triggers. It can also offer you increased coping skills and open your eyes to new ways of dealing with situations that you may not have been aware of before. Therapy can offer problem-solving skills, provide support, and help you work through life changes, allowing you to see your circumstances as a personal growth opportunity instead of a burden or obstacle.
Some specific skills therapy can provide are:
- Emotional management. You may not be able to control the feelings that come in to your mind but you can learn how to move through those emotions and choose which are worthy of action.
- Coping mechanisms to allow you to work through situations which typically cause you anxiety, fear, or avoidance.
- Stress-management techniques to apply to deal with stress within your everyday life, in your personal life as well as your work life.
- Skills and techniques to help you better navigate relationships, or to work through relationship troubles.
- Problem-solving skills for you to enact when you encounter issues which may typically have caused you to shy away or back down, such as social situations or public speaking.
- Improving self-love, self-confidence, and body image.
- Improving communication, listening, and the ability to speak up for yourself.
- Understanding your own skills, strengths, and positive attributes and learning to quiet your inner negative critic.
- Finding a resolution to the issues that originally led you to therapy, such as having panic attacks, or being unable to sleep.
While the decision to begin therapy is an individual choice, in many regards it may be helpful for everyone. There are a wide variety of reasons to begin therapy ranging from diagnosed mental illnesses to help to manage daily stress or a transitional period of your life. Your individual circumstances will set the tone.
What to expect on your first visit?
Your first therapy session has two main goals:
1. Assess your circumstances
We will take the first meeting to assess your current circumstances. While my areas of expertise may be consistent with your reasons for coming to therapy, we will need to address specific areas that are unique to you and your current circumstances. From there, we will be able to better determine what type of therapy is right for you and what your priorities may be. Sometimes, I may provide you with a series of actions to do outside of our therapy sessions, such as practice a certain technique, or read a specific book. It is important you take on an active role in your healing.
2. Begin a relationship
Our first session will be more like a two-way interview. I'll get to know you, and you'll get to know me. I will ask you questions to help me better understand your primary issues and concerns, as well as your history in terms of other events in your life, family, childhood, and career. However, you are welcome to ask questions too. In order for therapy to be successful, it is imperative we establish a client/therapist relationship that is supportive and honest. In fact, it is the nature – and the quality – of our relationship that will determine the success of your overall therapy goals. The success of the 'meeting of the minds' with your psychotherapist is the most accurate predictor of a positive, healthy outcome to the hard work you put in towards your happiness. As such, each client/therapist relationship will be unique but certain values and themes are true for all sessions, and you can expect the following:
- You can expect to be treated with compassion, empathy, respect, and understanding.
- You can expect to be heard and have your interpretation of what you are currently experiencing valued.
- You can expect to receive knowledgeable and scientifically backed techniques and information to assist you in overcoming your mental health-related struggles.
- You can expect to arrive in a safe, supportive, and confidential space.
- You can expect to receive real strategies and techniques you can use to enact positive changes in your life.
Is therapy confidential?
As a general rule, all therapy sessions are confidential and anything you discuss with your therapist will remain between the two of you unless you request otherwise. This is as per protection rules by law, which all therapists legally need to follow, and no information from the session can be disclosed without prior written consent from the client.
There are exceptions to this law however, and the therapist can disclose information from the session to legal authorities or appointed persons if any of the following are true:
- The therapist suspects abuse to a child, dependent adult, or an elder, or are made aware of domestic abuse. These situations all require the therapist to notify law authorities immediately.
- If the therapist suspects an individual has caused or is threatening to cause severe bodily harm to another person, therapists are required to report it to the police.
- If an individual intends to harm himself or herself, expressing to the therapist, for example, plans for suicide. While the therapist will attempt to work through this in the therapy session if it appears to be unresolved or the client does not cooperate, additional action may need to be taken to ensure the safety of the client.


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Family Theatre Company Presents Energetic Music Man at Westchester Broadway TheatrePublished: Thursday, July 12, 2012 7:00 am By: GEORGE J. DACRE
With great choreography and wonderful costumes, the production of The Music Man is full of the great musicals energy. The production stars Scott Bierko as Professor Harold Hill, who found trouble in River City when he proclaimed that the Indiana town needed a marching band, music, and instruments, then told them he would sell the instruments to the citizens. Directed by John Fanelli and starring dozens of youngsters eager to break into show business, The Music Man is the perfect vehicle for the enthusiastic cast that appears on the WBT stage.
They will be performing through July 29 with special Saturday matinees. Bierko and Victoria Lauzon, who plays Marian the librarian, make beautiful music together and are backed up by a very competent orchestra led by keyboard player and conductor Kurt Kelley. The show moves well and builds to the great climax when the band produced by Professor Hill excels on the musical number 76 Trombones.
A great quartet, coached by Sheldon Becton, play the role of the Buffalo Bills (in the movie) and make a wonderful harmony. The River City Dancers, Danny Hammond, Nicole Sartor and Carrie Silvernail are outstanding, as were the Teen Ensemble and the Childrens Ensemble.
The Music Man is a fun show. The music, which includes Till There Was You, 76 Trombones, Lida Rose, Gary Indiana, Shipoop, and Pick-A-Little, is professionally presented with keyboards, reeds, violin, trombone, and drums, which are performed by the onstage orchestra. The orchestra made last Fridays press night a great musical evening of theatre. I rate The Music Man at Westchester Broadway Theare in Elmsford Three out of Four Stars!
Tickets at 914-592-2222 or broadwaytheatre.com.


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BMW Says "No M1 on the Horizon"....Hints at "S" Variant for the 135i
Just like the 335is, introduced late last year, the 135is will be powered by the updated and retuned N54 Twin Turbocharged 3.0 liter inline-6. Peak power output has increased to 320 horsepower and 332lb/ft at the flywheel, up from 300 horsepower and 305lb/ft in the previous generation. As we would expect, a 6-speed manual transmission is the standard equipment, with a 7-speed dual clutch transmission available at an additional cost.
Initial testing conducted on the 335is has yielded a 0-60mph time of right at 5 seconds. Weighing in about 300lbs less, the 135is should be able to make the same sprint in about 4.9 seconds. A high performance suspension should help with the launch and the new aero kit for the "s" variant will add an aggressive touch to the exterior.
While no official pricing has been release, it should hover just under the 335i coupe.Auto Blog


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How to Solve Rand McNally Dock will not Install Problem?
Rand McNally is a respected company in the industry of GPS devices. There are several other companies that deal with these devices but the features of the devices offered by Rand McNally make this company reach heights. To make the use of these devices in a proper manner, it is very important for the person to regularly perform Maps and GPS device update. The process of update is rooted in the Rand McNally Dock software. There are times when the Rand McNally won't Install in the system of the user and eventually it will create a lot of problems in the working of the GPS device.
The dock is the medium through which all the updates process in a Rand McNally device is performed and to make proper use of the device, proper usage of the Dock is a must. When you face a problem of installation, it indirectly affects the usage of the GPS device as it stops the users from performing the GPS and Maps update.
We are going to talk about some reasons due to which this problem can occur in front of the users and along with that, we would also discuss some steps that would help the users incomplete the Rand McNally Dock Installation process in an easy way. Let's talk about the steps that would help you Install Rand McNally Software at first and then we would move on to the reasons behind this problem.
Steps to Install Rand McNally Dock
- At first, you need to turn off the Rand McNally device and allow it to power up properly.
- After that, you must connect the device to your system using a USB cable.
- Turn on your Computer for starting with the process.
- You must download Rand McNally Dock software using the internet and make sure that the download completes successfully.
- Now, you must Run Rand McNally Dock in your system.
- You will notice a window on the computer screen that would say, Welcome to Rand McNally Setup Wizards.
- Click Next and you will see a window that would contain the License Agreement of the software.
- You must click on "I Agree with" Button and tap on Next to continue with the installation.
- If you need to end the installation process then you must tap on "I Do Not Agree" and Click on Next.
- As the process continues, you must follow the instructions that are listed on the window.
- Complete the process of installation properly and click on Finish Button once the installation process is successful.
- You will notice that the Rand McNally Dock icon will be visible on the screen of your system either on the Desktop or on the system tray.
- Now, you need to register yourself as a new user in the software.
- You will be guided by the Rand McNally Dock with a demo and you will come to know about the process of using the Rand McNally Tool, Dock and Maps in a proper manner.
So, these are some of the steps that would help you in the process to Install Rand McNally in your system. Once the installation has been completed, you will be able to Update GPS and Maps in a proper manner.
Now, coming back to the Rand Mcnally dock repair tool if you are facing any problem while downloading. There are several reasons due to which this problem can be faced by the users of the GPS and here we have listed some major reasons for the problem.
Reasons for the problem.
- The major reason which is seen by the users of the product is due to the wrong installation of the software.
- Another reason for the problem is when the software does not recognize the device it is being installed into.
- If your system does not have the required configurations then you may face this problem.
- Another major reason for the problem is when the system is affected or corrupt.
So, these are the reasons due to which the Rand McNally Dock won't Install in your system. We advise our readers to go through the reasons and identify which of them is creating problems for them. Once you find out the problem, solve it and follow the steps that have been listed for the proper installation of the software.


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Macy's CEO Lundgren says online shopping helps drive in-store sales
The mutually beneficial relationship between online and real world shopping works both ways. Some customers see items in the store and choose to buy them online to take advantage of the shipping options. Whatever the motive, online sales are growing fast. "We're going to do close to a billion dollars in online sales this year and it's growing at 16 percent year-to-date," Lundgren said in an interview with DailyFinance at the company's Herald Square store in New York City on Black Friday.
U.S.consumers are expected to spend $44.7 billion on holiday online shopping this year, an 8 percent increase from 2008, according to Forrester Research.
The increase in online sales has helped Macy's improve its retail business because the company is better in touch with consumer demand, Lundgren said. "This multi-channel shopping is here to stay and it's just going to grow in importance over time."
Lundgren, who has been to over 100 Macy's stores in the last 100 days, said he learns about the business by talking to customers and his best retail workers. "I don't learn sitting in my office what's happening with consumers," he said.
Today, on the much ballyhooed Cyber Monday, many shoppers are sitting at their desks, surfing the Web and buying online. Lundgren said Macy's plan is to give customers "the value that they're looking for."
Anthony Massucci is a senior writer and columnist for DailyFinance. You may follow him on Twitter at hianthony.


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It can be hard for parents to watch their kids' desires for electronics outweigh their desire for other "more educational" things. As a mom of a 2-year-old, I'm growing increasingly more aware of the effects technology has on my daughter and do my best to ensure she has a healthy balance of "screen time" and non-screen time. Luckily, for parents who are concerned, there are plenty of amazing apps that spark creativity in children, ensuring that your child isn't completely shutting off their minds during screen time.
In fact, an article from Forbes notes the absurdity of the claim that children can't learn anything beneficial from playing with electronics or watching movies. While the frequency in which your child is in front of a screen is certainly something to take in to consideration, even more important is the question of what your child is consuming during screen time.
Letting your child play with apps that will stretch their imagination, teach them new skills, and show them pieces of history are definitely worth their weight in gold. And while an app about playing outside certainly can't replace the real thing, they're an amazing tool for learning— and lets be honest, keeping kids entertained on road trips.
1. Hopscotch For Iphone
With the popular app for iPads having over 4 million downloads, Hopscotch (Free) makes programing software easy for kids and teens, letting them design their own games, art, and more.
2. Sago Mini Music Box
Best for ages two to four, Sago Mini Music Box ($3) teaches kids control the music no matter where they touch on the screen.
3. PicsArt For Kids
PicsArt For Kids (Free) lets toddlers draw, scribble, and create silly scenes with the touch of a finger with a huge selection of colors, shapes, and pictures. And it's free!
4. My PlayHome
My PlayHome ($4) is essentially a virtual doll house where your child can dress, rearrange, and play with the dolls just like she would with a physical doll house. All of the fun and none of the tiny pieces.
5. Create A Car
If your kiddo is into cars or trucks, this app will be perfect. Create A Car ($0.99) lets children design and drive their very own vehicles.
6. Toontastic
This interactive app lets children create their own cartoons. In Toontastic (Free) children can draw, animate, and tell their own stories — the perfect outlet for a budding storyteller.
7. My First Classical Music App
My First Classical Music App ($5) teaches your child about different instruments, genres and composers, complete with dancing and singing animals. It's recommended for kids ages four and up.
8. Art Class With Dr. Panda
Art Class With Dr. Panda ($3) lets children break out their virtual scissors, crayons and paint and make masterpieces as beautiful as if they were in art class.
9. Toca Hair Salon Me
Despite its odd name, your kids will love this app. Toca Hair Salon Me ($3) lets children upload their own photos and then give them ridiculous hair-cuts and makeovers. Not necessarily educational, but most definitely creative.
10. The Daily Monster Monster Maker
Your child can hand make their own monsters in this creative app best suited for kids four and up. The Daily Monster Monster Maker (Free) features a large selection of pre-drawn "monster parts" kids can combine, or gives them the option to draw them on their own and send them to a friend.
11. MoMA Art Lab
The MoMA Art Lab (Free) is an interactive design space for kids to work with shapes, colors, and sound to produce unique works of art.


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Catexel - Opportunities in coatings´ quest for going cobalt-free - now and in the future
Thursday, 16 February 2017
Perspectives on the status of the alkyd coatings market
The challenge of replacing cobalt for the global coatings market remains.
Paint makers need more options in the face of regulatory reform.
Learn how R&D and the supply chain can support the requirement for fully optimised systems that satisfy commercial, operational, sustainability and corporate responsibility concerns.
In the two-part paper, Catexel´s CTO Dr. Ronald Hage - alongside independent market experts Dr. Hugh Gibbs and William Weaver - addresses:
- Key market challenges and predictions
- Crucial developments to-date
- Relative pros and cons of current cobalt alternatives
- Likely future innovation sources
For downloading this whitepaper, please log in.


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5 Probate Emergencies
If you haven't ever faced a probate emergency you probably wonder what kind of situation would require an emergency probate proceeding. It doesn't happen everyday. Most people have their affairs in order or they just don't have Murphy or Mayhem in their lives. There are five situations when you realize you have a probate emergency:
(1) the life insurance policy is locked in a safe deposit box and no one has access or authority to access the box and the funeral home or cemetery needs to be paid;
(2) the will or trust is locked in the safe deposit box and business associates, vendors, suppliers, or family members need to know who the executor is
(3) a will contest has broken out and disputing potential beneficiaries want to secure the deceased person's assets and property;
(4) the deceased person's business will suffer immediate harm unless an administrator is appointed to run the business; and
(5) a lawsuit must be filed or answered within time limits or be barred by statute of limitations.


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British tourist raped in Central Delhi hotel
A British tourist, staying at a hotel in Central Delhi, has been allegedly raped by a Polish national.india Updated:
A British tourist, staying at a hotel in Central Delhi, has been allegedly raped by a Polish national.
The Polish citizen, identified as Dominique Rafel, has been arrested after the incident which happened earlier last week, police said.
The victim was known to the accused as they were staying at nearby hotels in Paharganj area, a favourite among tourists because of the availibilty of a string of budget hotels.
The two had gone to a nightclub on February 7 and had drinks together after which Rafel allegedly raped the 28-year- old after taking her to his room.
She filed a complaint against him the next day and he was soon arrested. He was produced before Tis Hazari court, which remanded him to judicial custody.
Police said it has informed the Polish Embassy and British High Commission about the incident.


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Pilot Penguin wrote:Hi Jetman
I have to agree with you, this aircraft is impressive. But please keep in mind that isix did not do this entire aircraft from scratch. Isix expanded the original 777-200ER and tweaked the FDM in order to make the
Pilot Penguin
P.S. I'm not trying to downgrade your work or anything isix, so if I came across that way, I apologize. I am just simply trying to get the facts straight.
kyokoyama wrote:The Gold Bird thing looks big, but still, that's an amazing livery...!
Rick Ace wrote:I don't want to be impatient or rush anyone... but... Any idea when the first release it?
isix wrote:Ok, here the alpha :
http://www.4shared.com/file/luggqV5T/77 ... 1115a.html
Users browsing this forum: Richard and 2 guests


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So beautiful and creative! I love colored glass, but have never known what to do with my hoarded pieces. :)ReplyDelete
this is brilliant! my husband is always finding really pretty stones I never know what to do with this would be perfect. Thanks for sharing!ReplyDelete
mm super excited to make some for the living room!ReplyDelete
that's super creative!ReplyDelete
<3 Sharmaine Ruth
Cute idea!ReplyDelete
ok. i lOVE this. so much.ReplyDelete
I paint my plain circular cork coasters. But your idea is cool too.ReplyDelete
Love these! The paint and glass chips together are really prettyReplyDelete
Gorgeous DIY Kensey, what a great idea and you really a great job executing this DIY!ReplyDelete
adore these! Thanks for sharing. so creative.ReplyDelete
these are gorgeous. Such a great idea!!!!ReplyDelete
wow, very cool!ReplyDelete
I love this, it looks beautifulReplyDelete
I always can't wait for your DIY, you're a genious! xReplyDelete
it's soooooooo awsome!!!ReplyDelete
Soooo pretty!ReplyDelete
This reminds me of a beach house :) Come to think of it almost everything reminds me of a beach house these days...LOLReplyDelete
Great idea Kinsey, I love how it turned out!
Have an amazing week!
This idea is probably one of the best I've ever saw! I LOVE it!ReplyDelete
Really pretty and such a great idea! I NEED to try this out! <3ReplyDelete
- Anonymous4:59 PM, June 26, 2012
Love this idea!ReplyDelete
- Anonymous8:20 AM, June 27, 2012
Such a nice idea! :)ReplyDelete
The Jade necklace;)ReplyDelete
I follow Laura's lovely blogReplyDelete
And I've liked her page on fb;)ReplyDelete
These are so fun :) It reminds me of a project I did in high school with glass shards in different colors and shapes.ReplyDelete
I would say a perfect use of waste glass.. Awesome work done...ReplyDelete
How to Glue Glass
That DIY Stained Glass Coasters is so awesome. This must be included with my project plans next year. I have a passion for DIY projects.ReplyDelete


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Sep 18, 2007
Available from:
Sep 18, 2007 | ISBN 9780767928519
From one of America's most beloved television actresses: A sharply observed, comedic novel about the flipside of fame—and a fresh take on Hollywood in all its outrageous, entertaining glory.On the outside, Kate Keyes-Morgan seems to have it all: a starring role on the hit television series Generations, a handsome husband who is also her manager, a stick-thin figure, and a career that, after a Kirstie Alley-style slide, is finally back on track. At the age of thirty, Kate knows what it's like to have the tabloids turn on you, and she never wants it to happen again. Thank goodness her husband, Hamilton, is there to manage her every move—even if the strict dieting and morning weigh-ins are becoming more than she can manage. But when Hamilton leaves her for another actress on Generations, the ruthless diva Sapphire Rose, Kate suddenly finds herself in charge of her own life. Now she must decide whether she's ready to stop playing by the Hollywood rulebook and discover who she really is. With the help of her wise-cracking friend and tough-love hairstylist, Paige, and Michael, a high-powered film agent who secretly dreams of becoming a novelist, Kate begins to question her role in an industry that venerates appearance, money, and fame above all else, and that applies ceaseless pressure on women to always be thinner, younger, and more beautiful, at any cost.From high fashion to on-set antics to the real-life whispers about celebrities that even the tabloids never tell, Courtney Thorne-Smith depicts the entertainment world as only an insider can. Yet beneath the glamorous facade lies a natural-born writer with a gift for comic timing and shaping memorable characters. If you've ever wanted to experience life in the spotlight or wondered what goes on behind dressing room doors, Outside In is your invitation into the glittering world of Hollywood's biggest stars.
COURTNEY THORNE-SMITH is best known for her roles as Alison on the primetime soap opera Melrose Place, Georgia on the Emmy Award-winning series Ally McBeal, and currently as Cheryl opposite Jim Belushi in the hit situation comedy According to Jim…. More about Courtney Thorne-Smith
"A remarkably charming, whimsical novel . . . Thorne-Smith has a knack for swift, funny dialogue." —BookPage"A satisfying slice of chick lit."—Entertainment Weekly"An incisive tell-all about diet- and thinness-obsessed Hollywood." —Kirkus Reviews
Author interview
Q: How did you start writing? Have you always been a writer?I've always kept a journal, and have endless pages containing the first few paragraphs of a short story or essay that has been rattling around in my imagination. For years into my acting career, I would hear people say "I'm a writer" and my heart would hurt with yearning. About 5 years ago, when I realized that my words were getting twisted in the interviews I did about body image, I decided it was time to put that yearning to work and find a way to speak for myself. Q: How does your experience as an actor tie in to your experience as an author? Are there any similarities between acting and writing?Absolutely! My acting has informed my writing in ways I could never have imagined. Obviously my ear for dialogue is well trained, and when I write, I can't help saying each character's words out loud, as if I were being hired to play them (quite a stretch with the male characters, but actors always feels like they can play anyone!). More importantly, however, is how my experience as an actor helped me to create characters. As an actor, I use common feelings and experiences to bring the roles I play to life. We have all felt happy, sad, loved, neglected, proud, and ashamed. It is in the combination and dominance of one trait or feeling over another that character and personality are illustrated.Q: What's the message about friendship that women can take from Outside In? Is there a Paige in your life?There are a few Paiges in my life–and they are all claiming her as a personal characterization! Paige is a combination of "my girls," which is how I refer to my tight group of girlfriends and my wonderful mom and sister. I believe that the love and support I have felt with these women has saved my sanity many times. There is nothing as soul-satisfying as starting a gab fest feeling weighed down by emotional pain, only to find yourself laughing about your problems a few short hours later. That's what my friends give me: honest, loving reflection balanced with humor. Q: When it comes to fame, do you think the Sapphire factor–the materialism, the phoniness, the childishness–has worsened or improved in recent years? Or has that facet of Hollywood always been there?Unfortunately, I think there will always be some people who consider a movie or television set their own personal ego-gratification playground. Having said that, I also see how the business is changing, with fewer jobs available. As a result, I see more and more talented, successful people who are no longer willing to waste time and money. It is truly a gift to have the opportunity to make a living in a creative field surrounded by creative people, especially in this unsure economic climate.Q: What are some of your favorite books? What authors do you appreciate and admire?Oh dear, I get asked this question all the time and it is so difficult to answer! I have been a voracious reader since I was about five years old and I have been impacted by so many books and authors. As a young girl, I read and re-read The Phantom Tollbooth, Harriet the Spy, and all of Judy Blume's books until they were as soft and worn as tissue paper. As an adult, I love to get lost in classics by Jane Austen or the Bronte sisters. I always know that I am in for a treat when Anne Tyler, Jennifer Weiner, Marian Keyes, Jane Green, or Diana Gabaldon comes out with a new book. And Anne Lamott holds a special place in my heart, both for her honest, relatable fiction and her wonderful book on writing, Bird by Bird. Q: In this book, you take on a subject that every tabloid exploits: the pressure that Hollywood places on actresses to be thin. What are your thoughts on this subject? How pervasive is the pressure to maintain an unreasonably low weight, and what do you think women can do to stay healthy in the face of this?First of all, there has been a lot of blame placed on "Hollywood" for the pressure women feel to be thin. Way before there ever was a Hollywood, women and men were expected to conform to whatever body type was fashionable at the time. I do understand that the constant onslaught of television and magazine images that we now face every day has made it more difficult for women to forge their own paths and create images of beauty that are healthy and supportive, but it is by no means impossible. For every painfully thin image we see, there is a photo of the stunning Salma Hayek, striking Scarlett Johansson, or my personal favorite, Nigella Lawson, the incredibly gorgeous and vivacious chef and lifestyle expert. My point is that we have more power than we realize in terms of where we put our attention and energy. As a side note, when the lists come out every year about which women's bodies men most admire, it is never the skinny-minnies. It is always fit, healthy women like Jennifer Lopez, Jennifer Garner, Jessica Biel, and the women I mentioned earlier. As individuals, we need to expend our precious time and energy getting whatever support we need to find a way of eating and exercising that supports our bodies and minds, not waste countless hours of the day focused on starving our way to a completely subjective and fabricated ideal.Q: This isn't just a Hollywood story–it's also about love and second chances. Is it particularly difficult to find love in Hollywood?I believe true love is difficult to find anywhere, which is why it is so precious. Being famous certainly brings its own set of issues, but so does any career or lifestyle. One of the reasons I like working in television is that most shows are filmed in Los Angeles, and I found location shoots to be very hard on my relationships. I'm sure that businessmen and women who travel a lot for work, or people who work night shifts find the same to be true. Basically, I think we are all trying to find balance. And love. And the perfect pair of jeans.Q: Who did you write this book for? Who do see as the audience for Outside In?Well, I literally wrote it for my girlfriends, emailing them pages whenever I amassed more than ten or fifteen, or whenever Hamilton and Sapphire did something extraordinarily ridiculous. I also wrote it for readers like myself: people who like to go on a journey of self-discovery with a character they can relate to, who hopefully makes them laugh a lot and cry a little along the way. Q: Any sequels in the works?As a matter of fact, there is. It starts up where Outside In leaves off, and follows Kate further along her journey, while delving much more deeply into Paige's life. I guess the characters aren't done with me yet!
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