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US 4966839 A Abstract The present invention provides a process for the determination of a bindable analyte according to the principle of heterogeneous immunoassay by incubation of a sample solution which contains the analyte with a labelled first receptor specifically bindable with the analyte and present in dissolved phase and a second receptor present in a solid phase which does not cross-react with the first receptor and can fix a complex which contains the analyte and first receptor, separation of the phases after incubation and quantitative measurement of the labelling bound to the solid phase, wherein there is determined the back dissociation velocity of the labelling bound to the solid phase into the dissolved phase and the quotients of the back dissociation velocity and measurement value are used as a measure for the correctness of the test result of the first measurement.
Claims(8) 1. Process for determining if the Hook effect occurs in an analyste containing liquid sample, comprising incubating an analyte containing sample with a labeled first receptor which binds to said analyte and a second, solid phase bound receptor which is not cross reactive with said first receptor and which binds to complex containing said first receptor and analyte, separating liquid phase from solid phase, measuring label bound to said solid phase at a first time after said liquid phase is separated therefrom, and at a further time after the first measurement the back dissociation velocity of said labeled first receptor is determined, and relating said back dissociation value for said sample to a control back dissociation value, wherein a decrease in back dissociation value for said sample as compared to said control is an indication of the occurrence of the Hook Effect.
2. Process of claim 1, wherein said further measurement comprises measuring label bound to the solid phase.
3. Process of claim 1, wherein said solid phase bound receptor binds to said complex of first receptor and analyte to form a three member solid phase receptor-analyte-receptor complex, said process comprising contacting said three member complex with a solution of a color forming reagent, removing said reagent solution, and determining said back dissociation value by taking said first and said further measurement in said reagent solution.
4. Process of claim 1, further comprising contacting said solid phase with a wash solution after taking said first measurement and taking said further measurement in either said solid phase or said wash solution.
5. Process of claim 1, further comprising adding a third receptor which specifically binds to a determinant of said analyte which is different from the receptor to which said first receptor binds wherein said second and third receptors bind with each other.
6. Process of claim 5, wherein said third receptor is a monoclonal antibody, said first receptor is a Fab fragment of a monoclonal antibody, and said second receptor binds to the Fc portion of said third receptor.
7. Process of claim 1, wherein said first and further measurements are taken over a time interval ranging from about 20 seconds to about 60 minutes.
8. Process of claim 1 wherein at least one of said receptors is selected from the group consisting of protein A, immunoglobulin G, avidin, biotin, concanavalin A and lectin.
Description This application is a continuation, of application Ser. No. 926,641, filed Nov. 3, 1986 now abandoned. The present invention is concerned with a process and a reagent for the determination of a bindable analyte according to the principle of the two-side heterogeneous immunoassay. Two-sided immunoassays (sandwich tests) show marked advantages with regard to exactitude, sensitivity and rapidity as compared to competitive immunoassays. When the concentration of reagent, material used for the assay, or sample varies, the measurement value for the assay reaches, and passes through a maximum value. This effect is called the hook effect. For sandwich assays involving a solid phase, as analyte concentration increases, the measurement signal first increases, and reaches a maximum. As analyte concentration increases further, however, the measurement signal decreases. In this case, the same result can be brought about by two different amounts of analyte (see FIG. 1 of the accompanying drawings). This effect, which is dependent upon the amount of analyte, is called the "high dose" hook effect and limits the use of sandwich assays in spite of the above-mentioned advantages. Various reasons are given for the hook effect, such as heterogeneity of the antibodies present in the receptors or incomplete washing (see D. Rodbard et at., Immunochemistry, 15, 77--82/1978). Additionally, theoretical considerations lead to the conclusion that the hook effect must occur for all one-step sandwich tests. As one-step tests are meant all tests in which the analyte is brought to reaction with two receptors specific for it in the same solution and thereby or subsequently is fixed to an insoluble carrier material. This is in contrast to sequential assays where, after reaction of the analyte with a first specific receptor and fixing of the complex formed on to a solid phase, non-bound analyte is removed by washing the solid phase before reacting the complex with a second specific labelled receptor. In the case of the one-step sandwich test, the hook effect automatically occurs when both specific receptors are present in an insufficient amount with respect to the analyte so that only a part of the analyte is complexed with conjugate and also only a part of these complexes is fixed to the solid phase. All protein determinations with the help of a third"catching" receptor, involving complexing of two soluble receptors with the analyte and solid phase fixation via a further receptor, are also, in the above sense, one-step tests and display a hook effect. This can, inter alia, in the case of tumour markers, for example AFP, CEA, hCG, IgE etc., simulate results in the normal range in the case of already highly pathological samples. Therefore, for medical determinations, the use of the one-step sandwich process is frequently not used as a matter of principle. A recognition of such measurement results which simulate false values due to the hook effect is an important advance since all advantages of the onestep sandwich method (shortening of the reaction time, sensitivity, exactitude, etc.) could be utilized without the risk of a false result due to the hook effect having to be taken into account. K.L. Hoffmann et al. (Clinical Chemistry, 30 (9), 1499/1984) describe that a number of experiments have been carried out in order to prevent the appearance of the hook effect. Thus, for example, a stepwise analysis is suggested in which the analyte is first reacted with a receptor on the solid phase. Following this, the solid phase is then extensively washed, and this washing step is followed by a second incubation with soluble labelled receptor. Finally, the solid phase is washed a second time in order to separate the bound labelled receptors from the unbound ones. Furthermore, a reduction of the sample size, the carrying out of the test with various dilutions of the analyte or the use of only a small range of the calibration curve are suggested. The use of labels with low activity is also recommended, in order to allow the use of high concentrations of labelled receptors without the activity (radio, fluorescence, enzyme or colour forming activity) becoming too large. In addition, Hoffmann et al. (loc. cit.) describe a determination process for a one-step sandwich test in which the increase of the part lining to the solid phase of the labelled receptor is monitored kinetically and these values are fed into a computer, the reaction velocities then being calculated. Measurement results which have been brought about by the hook effect can thereby be differentiated from other values in that they display substantially smaller increases in complex binding. This process suffers from the disadvantage that extremely laborious and expensive computer processes have to be used in order to recognize whether a hook effect is present. Furthermore, this process is unsuitable for enzyme immunological determinations. Therefore, it is an object of the present invention to provide a process in which false measurement results brought about by the hook effect can be easily recognised without the inherent disadvantages of the known processes. These disadvantages include loss of sensitivity and loss of precision as well as increasing the amount of work, reagent and apparatus needed. Thus, according to the present invention, there is provided a process for the determination of a bindable analyte according to the principle of hetergeneous immunoassay by incubation of a sample solution which contains the analyte with a labelled first receptor specifically bindable with the analyte and present in dissolved phase and a second receptor present in a solid phase which does not cross-react with the first receptor and can fix a complex which contains the analyte and first receptor, separation of the phases after incubation and quantitative measurement of the labelling bound to the solid phase to obtain a first measurement, wherein there is determined the back dissociation velocity of the labelling bound to the solid phase into the dissolved phase and the quotients of the back dissociation velocity and measurement value are used as a measure for the correctness of the test result of the first measurement. In the usual sandwich test, after separation of the sample and reagent liquid, the solid phase-fixed sandwich complexes are determined via the amount of bound labelled receptors (radioactive, enzymatic, fluorometric or similarly labelled). Due to this separation and removal of non-bound labelled receptor, the equilibrium of the analyte-receptor binding is destroyed and a back dissociation of the sandwich components commences. This results in the presence of non-solid phase-fixed labelled receptor molecules in the liquid phase. The degree of back dissociation can be determined by determination of the labelling in the solid or separated liquid phase. As liquid phase, there can be used, for example, the wash liquid and, in the case of enzyme immunoassays, the substrate solution for the enzyme determination. In this substrate solution, according to the present invention, there is first determined the measurement value resulting at the point of time of the phase separation (colour signal) and subsequently the kinetics of the further colour formation are determined to measure back dissociation of the label. Since the liberation of an amount of label caused by back dissociation of the analyte-receptor complex proceeds approximately as a reaction of the first order, it is proportional to the amount of analyte and to the measurement signal associated therewith. Thus, the liberated amount of label can be expressed by a quotient alpha, which is the change of the measurement value after phase separation per unit time divided by the measurement value. Surprisingly, when a hook effect occurs, the quotient alpha is markedly changed in comparison with cases in which no hook effect occurs. Thus, with the help of the quotient alpha, it can be decided whether a measurement value obtained with the help of a calibration curve may be recalculated into an amount of analyte or whether the measurement value has been produced by a hook effect. In this case, it must be concluded that the sample contains an amount of analyte above the uppermost standard. In the scope of the process according to the present invention, analyte and receptor can, in principle, be all substance pairings which are bindable with one another. In this definition are included, from immunologically bindable substance pairs, and substance pairs which behave in an analogous manner. Besides the substance pair antigen-antibody, which first comes into consideration and is preferably used, (the term antibody here includes also known immunologically active fragments, not only of polyclonal but also of monoclonal origin), examples of substance pairs include protein A-immunoglobulin G, avidin-biotin, concanavilin A-lectin, as well as DNA-RNA (hybrid binding). All these substance pairs can be determined according to the principle of heterogeneous immunoassay (sandwich test) with one-step incubation according to the process of the present invention. As receptors, in the scope of the present invention, there are included the components of the above-mentioned specifically bindable substance pairs or derivatives thereof, especially the derivatives of bindable substances frequently referred to as conjugates, which are formed by covalent binding to a label molecule. An example of such a receptor, preferred in the scope of the present invention and which is present as conjugate, is an antibody or antibody fragment which is covalently bound with a labelling enzyme. However, instead of a labelling enzyme, there can also be used dyestuff molecules, colour forming components, especially fluorescent dyestuff molecules and the like as substance suitable for a detection system. Such a conjugate can also be radioactively labelled. Alternatively, the antibody itself can also contain radioactive atoms as label. In this case, the labelled first receptor consists, for example, of a radioactive antibody. The second receptor bound to the solid phase can also consist of a partner of the above-mentioned bindable substance pairs as such or a derivative thereof. The solid phase-bound second receptor serves, in the case of the sandwich assay, to fix the complex of analyte and first receptor to a solid phase and thus to make it easily separable from the liquid phase. For this purpose, the second receptor can bind directly with the analyte to be determined. In this case, the second receptor is directed against a determinant of the analyte other than that of the first receptor and, consequently, preferably consists of a monoclonal antibody or a fragment thereof. However, the second receptor can also be directed against another determinant of the complex of analyte and first receptor not present on the analyte itself. In this case, there is preferably also used a third receptor which is also specific for the analyte and recognises a determinant other than that recognised by the first receptor. Analyte, first receptor and third receptor then for a complex and, in this case, as second receptor, there is preferably used an antibody which binds with the third receptor. In an especially preferred embodiment, the first specific receptor contains an antibody fragment, especially a Fab fragment. and the third receptor consists of a complete antibody. In this case, the third receptor and the antibody part of the first receptor are monoclonal. The second receptor is, in this case, preferably directed against a part of the third receptor and especially preferably against the Fc part of the third receptor when the first receptor contains a Fab fragment. It is also possible to derivatise the third receptor and to use a second receptor which is directed against this derivatised place. The back dissociation velocity of the solid-phase bound label, which is important for the process according to the present invention, is determined by the binding strength between a receptor directed against the analyte and the analyte itself. According to a preferred embodiment of the present invention, the back dissociation constant for at least one analyte concentration is adjusted to an average value, i.e. one that is neither too high or too low a back dissociation velocity. If the velocity is too high, then the measurement intervals are, of necessity, ever smaller, which results in inexactitudes. Too low a dissociation velocity considerably increases the time necessary for carrying out the process and makes difficult the recognition of a reduced back dissociation rate such as occurs in the case of the presence of the hook effect. Since the process is preferably carried out in conventional automatic analysers, the capacity of the automatic devices can then no longer be fully utilised. As average back dissociation velocity, there is to be understood one which makes possible a measurement interval of from about 20 seconds to 60 minutes. A dissociation velocity in this range can be achieved by appropriate choice of the binding strength between a receptor and its binding partner, the analyte. For example by appropriate choice of the binding strength of the antibody, and/or by the binding strength-influencing additives in the reaction medium, such as surfaceactive substances and salts may be made. For this purpose, there are especially used detergents and/or chaotropic ions. Preferably, the various components and the intended measurement time period are so adjusted with regard to one another that at least, at one analyte concentration, within the period of time (measurement interval), at least 10% of the labelling bound to the solid phase again dissociates off. In the case of the preferred embodiment of the present invention with the use of an enzyme-labelled first receptor, after incubation of all components and separation of the phases, the separated solid phase is preferably incubated with a solution of a colour reagent for the determination of the labelling enzyme. Subsequently the colour reagent solution is separated from the solid phase and in the separated solution the back dissociation constant is determined by at least two measurements at a definite time interval. This embodiment is based upon the fact that, during the contact between the colour reagent solution and the solid phase, the total amount of the labelling enzyme bound to the solid phase and labeling enzyme possibly dissociated off during this incubation contribute to the colour formation. After phase separation, further color formation takes place which is only to be attributed to the dissociated-off labelling enzyme. The dissociated-off amount of the labelling enzyme (which corresponds to the dissociated-off amount of complex of analyte and first receptor) can, therefore, easily be determined by two measurements. According to a further embodiment of the present invention, after the first phase separation, the solid phase is incubated with a liquid phase, rather than a substrate solution (e.g. enzyme labeled). After a second phase separation, either the decrease of label on the solid phase or the label which has passed into the liquid phase. preferred for radioactive labellings and for colour and fluorescent labellings. The process according to the present invention can, in principle, be applied at all sandwich test systems. The only prerequisite is a measurable back dissociation of the labelled first receptor after separation of the non-bound first receptor remaining in the liquid phase. The peripheral conditions of the process according to the present invention are such that readily measurable back dissociations occur with and without the hook effect. The following Examples are given for the purpose of illustrating the present invention, reference thereby being made to the accompanying drawings, in which: FIG. 1 is a graphic representation of the dependency of the extinction (measurement value) upon the concentration of the analyte, the heavily marked part of the curve representing the calibration curve for the concentration determination of the analyte; and FIG. 2 illustrates an insert element for a centrifugal automatic analyser which can be used for carrying out the process according to the present invention. 70 mmole/litre HEPES/NaOH (pH 7.0) 154 μmole/litre sodium chloride 5 g./litre bovine serum albumin 5 mmole/litre chlorophenol red 7/8-galactoside (prepared according to Federal Republic of Germany Patent Specification No. 33 45 748) 2 g./litre Tween 20 (non-ionic detergent) As receptor 1 solution, there is used a monoclonal mouse anti-AFP antibody which recognises an antigenic determinant different from that recognised by receptor 3. An ascites liquid containing this antibody is mixed ad 1.8 M/litre with ammonium sulphate. The precipitate is taken up in 15 mM/litre sodium phosphate buffer (pH 7.0) containing 50 mM/litre sodium chloride. The solution so obtained is subjected to a passage over DEAE-cellulose. The complete antibody is split up in known manner into Fab fragments. The Fab fragments obtained are coupled with β-galactosidase according to the method of E. Ishikawa, J. of Immunoassay, 4 (1983), S. 209-327. Sheep anti-mouse Fcγ antiserum is, as described above under 2), purified and also subjected to a passage over DEAE-cellulose. As receptor 3, there is used a monoclonal mouse anti-AFP antibody which recognises a determinant other than that recognised by receptor 1. An ascites liquid containing this antibody is mixed ad 1.8 mole with ammonium sulphate. The precipitate is taken up in 15 mmole/litre sodium phosphate buffer (pH 7.0) containing 50 mMole/litre sodium chloride, solution thus obtained is subjected to a passage over DEAE-cellulose. (1) 40 μl. of a solution which, per litre, contains 100 mMole sodium phosphate (pH 7.3; 37 � C.), 2 mMole magnesium chloride, 9 g. sodium chloride, 5 g. bovine serum albumin, 5 mg. anti-AFP monoclonal antibody from mice (receptor 3 solution) and 2000 U anti-AFP antibody (mouse) Fab fragment β-galactosidase conjugate (receptor 1 solution); activity determined with o-nitrophenyl-β-galactosodase at 37� C., is applied dropwise to a fleece which consists of commercially available polyester paper. It is subsequently dried at ambient temperature. This fleece is stored at 4� C. and at a relative atmospheric humidity of 20% until it is used. On to a cellulose fleece, after cyanogen bromide activation (see Federal Republic of Germany Patent Specification No. 17 68 512), there are fixed sheep antibodies against the Fcγ part of mouse antibodies (receptor 2 solution), whereby, per g. of fibre material, here are made available for fixing 10 μg. of antibody. Uncoupled antibody is removed by washing and the fleece is gently dried at ambient temperature. The fleece thus obtained is stored analogously to reagent carrier 1. The determination with the help of these two reagent carriers 1 and 2 takes place with the device described in Federal Republic of Germany Patent Specification No. 34 25 008 for the carrying out of analytic determinations (see FIG. 2). This describes a rotor insert element for a centrifugal automatic analyser which consists of a plurality of reagent fields in combination, each of which contains an absorbent carrier material impregnated with a particular reagent, at least one mixing valve chamber and a measurement chamber which together form a sample liquid transport path which passes from radially inwardly to radially outwardly when the rotor insert element is fixed to the rotor and further has at least one further chamber for the reception of a liquid and a transport path which passes from this chamber to the measurement path and is at least in part identical with the sample liquid transport path. The sample liquid transport path thereby passes from a sample reception chamber (p) via a chamber (a) filled with an absorbent material containing buffer, a chamber (c) and a first valve chamber (Vkl) arranged between the chambers (a) and (c) to a second valve chamber (Vk2) and from this, via a chamber (d) and via a reception chamber (AK) to measurement chamber (K). For the reception of a second liquid, there is provided a substrate chamber constructed as a pump chamber (PK) which has a substrate application port (S); and which is connected with the second valve chamber (Vk2) via a dosing unit consisting of a dosing chamber (DK) and capillar (Kap) and an overflow chamber. FIG. 2 of the accompanying drawings shows schematically the rotor insert element used. Reagent carrier 1 is placed on field (c) of the rotor insert element and reagent carrier 2 on field (d). 40 μl. of sample are thereby pipetted through an opening on the upper edge directly on to the field (a). The sample is diluted 1:5 (v/v) with 0,9% aqueous sodium chloride solution, 270 μl of substrate solution are pipetted into chamber PK. By means of an appropriate centrifuging programmed, in which high speeds of rotation alternate with stopping, the sample and the substrate solution are then conveyed in the direction of the separation matrix and cuvette. In the course of the programmed, the receptors 1 and 3 are thereby eluted by the sample liquid from field (c) and the homogeneous mixture is subsequently brought to reaction. On field (d}, the complex formed is bound to receptor 2. The transfer of the sample from field (c) to (d) takes place within a very short space of time. The substrate solution is divided up by the dosing chamber DK into portions, the first of which serve for washing out excess, non-complexed conjugate. The β-galactosidase activity bound to (d) b, complex formation is proportional to the amount of AFP present in the sample. This activity is determined with a further portion of substrate, the substrate thereby being reacted in a 5 minute reaction to give coloured products. The colour formed and the further colour development/ minute in the liquid phase are measured in the cuvette at 600 nm. Under these conditions, the following results have been obtained: __________________________________________________________________________AFPIU/ml. 0 60 135 200 50,000 100,000 200,000 500,000 1,000,000__________________________________________________________________________measurement 274 1516 2783 5363 >8.000 6587 4670 2782 2069value (mE) ##STR1## 19 131 217 305 -- 48 40 30 32α = 6.9 8.6 7.8 5.7 -- 0.7 0.9 1.1 1.5increase/measurementvalue � 100 .BHorizBrace. hook effect__________________________________________________________________________ All measurements were carried out at 600 nm at a layer thickness of 0.3 cm. and recalculated to a layer thickness d=1cm. The procedure described in Example 1 is used but with the following differences: (a) as receptor 3, there is used a monoclonal mouse anti-IgE antibody directed against human IgE; (b) as receptor 1, there is used a further monoclonal mouse anti-IgE antibody directed against IgE which, however, recognises an antigen determinant different from that recognised by receptor 3; (c) in the preparation of reagent carrier 1, 12 mU/test of receptor 1 solution are used; and (d) the sample is diluted 1:3 with aqueous sodium chloride solution. Under the appropriate conditions, the following results were obtained: __________________________________________________________________________U/ml. 0 52 90 182 278 383 4891 12,2001 29,750 64,500 118,000__________________________________________________________________________measurement 88 604 1098 2050 2898 3646 4073 6445 4573 3151 2230value (mE)increase2 4 34 60 101 127 146 150 68 65 98 38(mE/min) ##STR2## 4.5 5.6 5.5 4.9 4.4 4.0 3.7 1.1 1.4 1.5 1.7 .BHorizBrace. hook effect__________________________________________________________________________ 1 at concentrations from about 500 to 12,000 U/ml., the photometer measurement range was exceeded 2 all measurements took place at 600 nm with a 0.3 cm. cuvette and the extinctions were subsequently recalculated to a 1 cm. light path The last three columns show values which are brought about by the hook effect and cannot be used for a quantitative determination of the analyte. Microtiter plates (MTPs) are coated, per cup, with 200 μl. of a solution of a monoclonal mouse anti-AFP antibody receptor 2 solution in a concentration of 1 μg. of protein/ml. in buffer A (20 mM carbonate buffer (pH 9.6)) for 1 hour at ambient temperature. Subsequently, non-specific binding phases are saturated by post-incubation of the MTPs with 300 μl. of buffer B (50 mM potassium phosphate buffer (pH 7.5), 154 mM sodium chloride, 1bovine serum albumin and 1 g./litre Tween 20) for 30 minutes at ambient temperature. After washing with 300 μl. of buffer C (100 mM HEPES/NaOH (pH 7.25), 154 mM sodium chloride, 2.5 g./ litre sheep normal IgG, 0.5 mM magnesium L-aspartate and 2 g./litre Tween 20), there are introduced. per cup, 50 μl. of sample and 200 1 of an enzyme-antibody conjugate (receptor 1 solution; β-galactosidase conjugate of a Fab fragment from a further monoclonal mouse anti-AFP antibody which does not hinder antibody from receptor 2: 0.5 U/ml.: measurement of the activity with o-nitrophenyl-β-D-galactoside at 37� ) in buffer C and incubated for 1 hour at ambient temperature. Subsequently, the liquids are discarded from the MTP cups, the cups are washed with 300 μl. buffer B and thereafter incubated with 250 μl. of substrate solution (5 mM chlorophenol red μ-D-galactoside, 70 mM HEPES/NaOH (pH 7.0), 154 mM sodium chloride, 3 /litre bovine serum albumin and 2 g./litre Tween 20) for 2.5 hours at ambient temperature. After the epiry of this incubation time, 100 μl. are removed from each cup of the MTP and transferred to the cups of a further MTP. Immediately thereafter, there takes place an absorption measurement (measurement t1) by means of a commercially available MTP measurement apparatus (=570 nm, bichromic correction at 630 nm) and after a further incubation period of 1 hour at ambient temperature there takes place a second absorption measurement (measurement t2) under the same conditions as measurement t1. The absorption difference between measurement t1 and measurement t2 is proportional to the amount of enzyme transferred with the substrate solution. The results of a typical experiment are given in the following Table: ______________________________________(AFP)inIU/ml 1.5 10 40 156 625 2,500 10,000 40,000______________________________________mEt1 67 419 1720 >.sup.(2) >.sup.(2) >.sup.(2) 1643 485mEt2 - 32 171 540 -- -- -- 141 51mEt1α.sup.(1) 478 408 314 -- -- -- 86 105______________________________________ The last two columns show the presence of a hook effect. (1) ##EQU1## (2) above the photometer measurement range (2000 mE). Patent Citations
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Seen this evening, about 1800hrs on junction of redding lane and shaftmoor lane, hall green I thunks ??? Looked like it was you Nilzy??? Was it?
What you doing in the ghetto dude? It wasnt me tho today, however there are a couple of other ones around now in the hood...
lol ghetto... was picking up some second hand stuff form my DSLR. is that your usual hunting ground then dude? lol
YO YO!!! I am seen about there sometimes, not too often though.... What were you getting for your cam? Was it from a shop or privately?
Private advert for a lens lol. Now found it doesn't work properly on my cam oh well. Should have researched first... EBay here I come.


text:
While searching through my teachers old patterns i encountered a romper that i absolutely loved (: 1986 Pattern from Esprit. I changed it a bit to fit me better and made it shorts instead of pants
Navy Broadcloth
Navy thread
6mm wide elastic
Aug 24, 2010, 07.25 PMby sibeli
Stunning! Love the ruffle detail in the front.
Aug 15, 2010, 05.31 PMby tiger-lily
I don't usually like rompers, but this is very cute! I really like the bodice. I think the pattern would make a cute top, too.
Aug 15, 2010, 08.22 AMby raving
Adorable, I love playsuits and this one is particularly sweet! :)
Aug 14, 2010, 12.50 PMby mema79
this is very stylish!
Aug 14, 2010, 11.51 AMby tamie3
i love this!!…it pretty
Aug 14, 2010, 05.56 AMby loriquesada
Looks GREAT!
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text:
a lot of relics, multiple layers, the thing is if you love the mosaics, you'll love this place, you'll love the city,..
Beautiful mosaics, some of them preserved quite well. One can see the layers of history. Ruins of ancient Roman temple are visible below the Church of Virgin Mary.
It's worth hiring a tour guide for this site, considering the entrance fee is close to nothing. When you walk in to see the mosaics they seem very dusty and unappealing but as soon as our guide splashed water onto the tiles, the colors were very vibrant and the map was easier to read. A lot of history in this... More
A lot of mosaics and images of how this place looked like in the "good old days". In this case during the Roman times.
There was a lot of interesting things to see at this little park. Amazing mosaics as well as a roman road, a church crypt and byzantine house foundations. It's nice that it is so small because you don't have to worry about museum fatigue ;-)
This park you can reached by walking distance from Georges Church. Pay 2 JOD for entrance fee and awesome!
I didn't hire a guide, and although there are little notices on the walls and I had a guide book, I couldn't get the 'whole story', which would have been more interesting. the mosaics were well preserved and the area interesting
The Archeological Park is only a short distance from St Georges Church. The mosaics contain remarkable scenes from Greek and Roman mythology from various buildings and churches in and around Madaba. Take note also of the beautiful mosaic images of birds and flowers. Running diagonally through the park is a substantial length of a magnificently wide paved Roman street with... More
The Medaba map gives evidence of the places and traditions of the Bible in Byzantine times and has led to recent archeological finds. Welll worth a visit and purchase of the explanatory booklet. The town itself is vety interesting and mosaics of all sizes and styles can be purchased here.
The park has some really amazing mosaics and the guide is very knowledgeable. Different epoches can be found and you can see the evolution of the art of making mosaics her.
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Python Programming...Need some help!!!
page: 1
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posted on May, 11 2004 @ 12:00 AM
I need some serious help with a computer science class. Here's my dilemma...
The sylabus originally stated that our final project (well not final project because this assignment should be ridiculously easy..) was originally supposed to be some basic coding and such with flash. I happen to be very proficient with flash (hence my taking this course as one of my easy ones.) However, unbeknownst to me until about a week ago, they changed the last project to making a simple program in Python (when I say simple I mean it should be a cake walk for anyone with basic knowledge of the language...see assignment below.) Because I was under the assumption that we were going to be doing flash, I didn't go to class and have now missed the steps and general knowledge required to complete the assignment. I know I could probably figure this thing out eventually, however, its due tomorrow by midnight and I have neither the time nor the energy to deal with this as I have another exam tomorrow that requires my utmost attention to prepare for. I was wondering if anyone could help me out. Heres the assignment and also some other stuff they gave us to get started.
As a matter of National security, we want to know when head TA Mike Viamari�s birthday and Easter occur on the same day: March 31st.
Your mission is to write a short program that tells us which years between 1900 and 2099 Easter falls on the 31st.
You will be supplied with a function called easterday(year). The value of the variable �year� is calculated into the day of the month that Easter falls on for that year.
A value between 1 and 31 is returned when the function is given with a year between 1900 and 2099
Heres the function easterDay they gave us:
def easterDay(year):
if year < 1900 or year > 2099:
raise ValueError, 'easterDay is only valid from 1900 to 2099'
year = int(year)
a = year%19
b = year%4
c = year%7
s = 19*a + 24
d = s%30
t = 2*b + 4*c + 6*d + 5
e = t%7
theDay = 22 + d + e
if theDay >= 32:
theDay = d + e - 9
elif theDay == 26:
theDay = 19
elif theDay == 25 and a == 16 and d == 28:
theDay = 18
return (theDay)
Basically I got the thing to work if I specify a specific year and it will spit out whether easter falls on the 31st for that year or not but where I'm stuck is getting the program to run through all the years automatically and spit out which particular years Easter falls on the 31st.
Much appreciation for anyone who can help me out of this small pickle.
[Edited on 11-5-2004 by hopblaze]
posted on May, 11 2004 @ 01:25 PM
Not sure if I am reading the question correct, but my answer would be this:
theYear = 0
for year in range(1900, 2099):
theYear = easterDay(year)
print theYear
I may have mis-understood the question, if I have - sorry. One comment though, if I did understand the question correctly, and my answer above is good for you - what the heck are you doing in a computer science scall when you can't work that out?
posted on May, 11 2004 @ 01:28 PM
sorry, ATS gets rid of tabs, so the code won't compile like that - u should know what tabs to put in, lol
posted on May, 11 2004 @ 01:32 PM
Or, to make it look better: (again, tabs are disposed off, so try and put the tabs in the right place):
theYear = 0
for year in range(1900, 2099):
theYear = easterDay(year)
print "year :" + theYear
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text:
Influence of metabolic stress on the inheritance of cell determination in the moss, Pottia intermedia
Article first published online: 2 JAN 2013
DOI: 10.1016/j.cellbi.2005.02.001
© The Author(s) Journal compilation © 2005 International Federation for Cell Biology
Additional Information
How to Cite
Lobachevska, O., Kyjak, N., Khorkavtsiv, O., Dovgalyuk, A., Kit, N., Klyuchivska, O., Stoika, R., Ripetsky, R. and Cove, D. (2005), Influence of metabolic stress on the inheritance of cell determination in the moss, Pottia intermedia. Cell Biology International, 29: 181–186. doi: 10.1016/j.cellbi.2005.02.001
Publication History
- Issue published online: 2 JAN 2013
- Article first published online: 2 JAN 2013
- Abstract
- References
- Cited By
- Aposporous regenerant;
- Pottia intermedia;
- Apogamy;
- Epigenetically-determined;
- The determined state;
- Cell inheritance
Epigenetically-determined apogamy in aposporous regenerants of the moss Pottia intermedia persists during vegetative propagation, the capacity of apogamy being inherited by individual aposporous protonemal cells. To test Bauer-Lazarenko's proposal that stable apogamy in mosses may be due to some self-replicating cytoplasmic factor, the effects of different metabolic stress treatments on the expression of apogamy have been tested. Chronic metabolic stress caused by long-term growth of autotrophic aposporous protonema on mineral medium with 0.25% of casamino acids and on Murashiga—Skoog (MS) medium with sucrose and phytohormones, as well as by transitory action of high kinetin concentration, have a much stronger influence on the expression of apogamy, than short-term stress treatments with RNase and Pb2+. Apogamy has been found to be lost stably, after prolonged growth on MS medium containing kinetin and ABA. The proposal that the capacity for apogamy is related to the release of aposporous protonemal cells from a putative factor for apogamy is discussed.


text:
Get ideas and inspiration, find easy to use patterns
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Kayhan Irani, New York-New Jersey regional coordinator for TIGRA, the Transnational Institute for Grassroots Research and Action. TIGRA is the umbrella organization coordinating the Western Union boycott.
A network of over 150 immigrant organizations across the country have launched a boycott of money transfer giant Western Union. The organizations accuse Western Union of charging immigrants exorbitant fees and non-standard exchange rates when they try to send money home. We speak with a coordinator for the Transnational Institute for Grassroots Research and Action, the umbrella organization coordinating the Western Union boycott. [includes rush transcript]
This is a rush transcript. Copy may not be in its final form.
JUAN GONZALEZ: Immigration reform might have died in the Senate earlier this summer, but the movement for immigrant rights is very much alive. Although Democratic hopefuls at the first Spanish-language presidential debate broadcast Sunday focused on the importance of border security, immigrant rights advocates aim to steer the debate in another direction. They are emphasizing the economic security of migrant workers.
A network of over 150 immigrant organizations around the country have launched a boycott of money transfer giant Western Union. The organizations accuse Western Union of charging immigrants exorbitant fees and non-standard exchange rates when they try to send money home. The boycott organizers say that Western Union's annual profit of over $1 billion comes from the fees paid on remittances sent by migrant workers to their home countries. Yet, Western Union, they say, reinvests only 41 cents for every $100 of profits back into these immigrant communities.
AMY GOODMAN: Kayhan Irani is the New York-New Jersey coordinator for TIGRA, or the Transnational Institute for Grassroots Research and Action, the umbrella group coordinating the Western Union boycott, joining us now in New York. Welcome to Democracy Now!
KAYHAN IRANI: Thank you.
AMY GOODMAN: Explain what you're doing.
KAYHAN IRANI: So the Transnational Institute for Grassroots Research in Action is working with, like you said, over 150 immigrants' rights groups in America and overseas. We aim to get justice for immigrants and allow them to build the wealth that they have in their communities. You have over $236 billion going around overseas from folks who are working in different countries, going to their families to support them for basic needs. This is not — no one's buying a second car with this money. No one's buying a fur coat. This is for healthcare. This is for housing. This is for adequate education. And these basic needs are being charged exorbitant rates. The people are really being taken advantage of. Their love, their commitment to their family is what's driving the profits of this company. And it's really sad to see Western Union completely withdrawing from any type of responsibility, and so we want to hold them accountable.
JUAN GONZALEZ: Now, my understanding is that Western Union has gotten into this position, because in many places in the third world, where a lot of towns don't have banks, but there will be three or four Western Union offices, so they are like the main transmitters of money?
KAYHAN IRANI: Absolutely.
JUAN GONZALEZ: When you talk about exorbitant fees, what are you talking about?
KAYHAN IRANI: So, for example, to send money home to someone, you can get charged between $15 and over $20. But the fees cost Western Union about $3 to $4. So the markup is huge over — again, over something that is not a luxury item. Sending money home is not a luxury item. Low-wage workers who need to support families in other countries, who have been torn apart from families, shouldn't be charged five — four and five times what it costs Western Union to send a remittance. It's completely ridiculous. And all the money goes back into Western Union. The billions of dollars of profits, $1.3 billion in operating profits that they made, doesn't go back to the community, it goes into the pockets of the corporation.
AMY GOODMAN: We asked Western Union to respond to the boycott. I want to play an audio recording of this statement from Daniel Diaz, a Western Union spokesperson.
DANIEL DIAZ: We are proud to provide fast, reliable, trusted customer service at competitive prices in a market with more choices than ever before. These choices have led to lower costs across our industry. We take pride in the role that Western Union plays in empowering communities everywhere, by making it possible for immigrants to support their families and give back to those they love most. Finally, we're proud of the direct engagements we've made with communities and partnerships we've formed to take action and support programs that empower individuals to become more self-sufficient. At Western Union, we're committed to serving our consumers worldwide.
JUAN GONZALEZ: Well, I'd like to get your response to that comment, but also I understand that even some international organizations like the World Bank and others have questioned these Western Union rates and the impact it's having on transmittals.
KAYHAN IRANI: Yes. In fact, the World Bank has said that if Western Union lowered their fees by a third, they would still be making plenty of money, that they would still have healthy competition and have a healthy profit margin over their competitors. The industry itself is inflated. The industry itself if being allowed to profit hugely, because no one has been raising their voice. This is the money. This is the love. This is the commitment of millions of people who have been ripped apart from their families, who have to go work in other countries and who work low-wage jobs. These are not rich people.
I went to an ESL class yesterday to talk to some folks out in Queens, and when I asked them, I said, "Are you guys millionaires? How do you see immigrants? Do you think immigrants are rich, immigrants have financial power?" And they all said, "No, no. We're not rich. We're poor." And I said, "Well, what about the $232 billion that you have? Because that's how much you sent over the last year."
AMY GOODMAN: I was reading a piece by Francis Calpotura from Foreign Policy in Focus called "Remittances: For Love and Money." Interesting figure: Studies show if money transfer fees were cut in half, 33 million people could be lifted out of poverty in the developing world. That's for money transfer fees overall.
KAYHAN IRANI: Right. That's the industry. And there you have it. It's completely ridiculous. It's really sad to see Western Union's comment about, you know, how they help immigrants build wealth and how they are giving back to the community and how they're very happy to empower the community, except when it comes down to it. Thirty-three million people could be lifted out of poverty, and they're fighting over change. Western Union reinvests 41 cents for every $100 of profit back into the community. That's abysmal.
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text:
League tables branded a 'nonsense' by private schools
January 29th, 2015
… Read the restSchool leaders dismissed this year's tables as a "nonsense", with many schools "caught unawares" by a shift in which qualifications are recognised.
… Read the restSir Michael Wilshaw told a committee of MPs on Wednesday that there was homophobic behaviour shown by children at Durham Free School when the school was inspected last year.
… Read the restClaims that hard-line Muslims tried to gain control of the schools have led to "big problems about leadership and staffing", said Sir Michael Wilshaw.
… Read the rest…Ofqual was sticking to its guns today over the science proposals, stating that they were "designed to invigorate the hands-on learning experience of students and equip them for a future in science".
… Read the restThe Mathematics for Life conference is tackling what is seen as the weakest of the core subjects taught in Wales.
… Read the restThe practice was revealed by Sean Harford, the inspectorate's national director for schools policy, during an education select committee hearing this morning…
He said: "What we don't want…is some practice I have heard where in some primary schools they are putting their pupil premium children on a table so they have a pupil premium group; that is not going to drive those children to achieve better, frankly."
He said similar practices were also taking place in secondary schools…
"Expecting teachers to put them on a table so they can identify them just in case we turn up, for example, is wrong," he added.
… Read the restThe head of Brighton College, Richard Cairns, will tell a conference tomorrow that plans to promote 'British values' after the Trojan Horse scandal in Birmingham should have done more to challenge homophobia.
… Read the restAlmost two-thirds of young people have used pornography to find out more about sex, and two-fifths say that it has helped them to understand sex.
… Read the restI am a passionate believer in the right of every child to an education and the opportunity of every child to make the best of themselves.
… Read the restCambourne Village College, which opened in Cambridgeshire two years ago and has 317 pupils, introduced tags to help children get to know each other, manage behaviour and help teachers.
… Read the restDebbie Evans, 55, who taught at South Wilts Grammar School in Salisbury for 23 years, struck up a sexual relationship with the girl in 2011, after the teenager fell ill at school and she supported her.
… Read the restDavid Tuohy, 83, who now lives in Oxford, took charge of Eccles Hall, a special school, in the 1970s.
… Read the rest…In a speech at the thinktank Politeia, Mrs Morgan: "No school should be exempt from promoting fundamental British values, just as no school should be exempt from promoting rigorous academic standards."
Mrs Morgan made an explicit link between indoctrinating primary school children and the attacks by Muslim terrorists on the satirical magazine Charlie Hebdo and a Jewish supermarket in Paris earlier this month.
… Read the restIn a strongly worded speech at the Politeia thinktank in London, Nicky Morgan said that dropping practical lab work from both A-level and GCSE results would harm the next generation of scientists.
… Read the restFigures suggest more than 1,600 did not have any pupils going to Oxford or Cambridge and about 335 sent none to a Russell Group university.
… Read the restOf youngsters educated in the state sector, 64% of Asian students and 62% of black students went on to higher education in 2012-13, compared to just 45% of white students.


text:
LUCKY STIFF, Starring Jason Alexander, to World Premiere at Montreal Film Festival, 8/22
LUCKY STIFF, the zany off-Broadway musical from Tony®-winners Lynn Ahrens and Stephen Flaherty, comes to the big screen! The film adaptation, starring Tony® winners Jason Alexander ("Seinfeld") and Nikki M. James ("The Book of Mormon," "LES MISERABLES"), Dominic Marsh ("Missing," "Doctors") and Pamela Shaw ("Swingers," "The Whistleblower"), will have its world premiere at the Montreal World Film Festival on Friday, August 22. It will compete in the World Competition category.
Attending the premiere will be the film's stars Nikki M. James and Pamela Shaw and producers Victor Syrmis and J. Todd Harris. The film will screen for the public on Friday, August 22 (9:30am), Saturday, August 23 (2pm) and Sunday, August 31 (9:30pm).
Also starring are Don Amendolia ("Law and Order," "Frasier"), two-time Tony® nominee Jayne Houdyshell ("Well," "Follies"), Kate Shindle ("Capote," "The Stepford Wives") and the late Dennis Farina ("Get Shorty", "Saving Private Ryan").
LUCKY STIFF is a musical comedy with a romantic heart. A young down-and-out British shoe salesman named Harry Witherspoon (Dominic Marsh) takes his dead American uncle (Don Amendolia), a murdered casino manager, to Monte Carlo for the best time of his life - a week of fun, dancing, gambling and sun. If the young man fulfills his uncle's request to the letter, he will inherit the $6 million left to him. If he doesn't, the money will go to the Universal Dog Home of Brooklyn. As Harry races from casino to nightclub to beach to bedroom with his dead uncle, he is chased by a desperate put-upon optometrist (Jason Alexander), his controlling, myopic, trigger-happy sister (Pamela Shaw), an avaricious French chanteuse (Kate Shindle), a mysterious Italian playboy (Dennis Farina), as well as a young woman from Brooklyn (Nikki M. James) dead set on getting that money for the dogs. Guns go off, disguises go on, champagne corks pop, nightmares come to life, romance blossoms, dogs bark, and everyone sings! It's a zany, frothy, colorful and fast-paced musical farce with a very happy ending.
LUCKY STIFF first premiered in 1988 off-Broadway at New York's Playwright's Horizons. A recipient of the Richard Rodgers Award and Washington's Helen Hayes Award for Best Musical, the musical has been performed all over the world including London's West End. LUCKY STIFF is based on Michael Butterworth's 1983 novel "The Man Who Broke the Bank at Monte Carlo."
LUCKY STIFF marks the feature film directorial debut of Tony®-nominee Christopher Ashley ("Memphis," "The Rocky Horror Show"). Victor Syrmis and J. Todd Harris produced the film. Tony® Award-winning writer Lynn Ahrens ("Ragtime," "Rocky") adapted the stage production for the screen.
LUCKY STIFF is set to be released in 2015. Watch the recently released trailer here: http://youtu.be/kXdT4FGiUnw
For updates on the film, follow LUCKY STIFF on Facebook.
LUCKY STIFF festival screenings will be held at the Cinéma Impérial (1430, Rue de Bleury, Montreal) at the following times:
Friday, August 22, 9:30am (for press only followed by press conference at 2pm)
Friday, August 22, 7pm (for public)
Saturday, August 23, 2pm (for public)
Sunday, August 31, 9:30pm (for public)
See Montreal World Film Festival's site for more details and to purchase tickets: http://www.ffm-montreal.org
About Author
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text:
Book your room at Extended Stay America - Washington, D.C. - Fairfax
Deluxe Studio Queen Bed non smoking
Max Occupancy: 4 guests (up to 3 children)Deluxe Studio King Bed Non Smoking
Max Occupancy: 4 guests (up to 3 children, 3 infants)Fairfax hotel with a seasonal outdoor pool
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Simply enter your Nectar card number when prompted. Your points will be credited 30 days after you return from your trip.*Offers may apply
Situated in the suburbs, this hotel is close to Ox Hill Battlefield Park, Fair Oaks Mall, and National Firearms Museum. Area attractions also include National Air and Space Museum Steven F. Udvar-Hazy Center and Walney Visitor Center.
Hotel Features
At Extended Stay America - Washington, D.C. - Fairfax, recreational amenities include a fitness center and a seasonal outdoor pool. The hotel also features a coffee shop/café and complimentary wireless Internet access.
Room Amenities
Flat-screen televisions come with premium cable channels. Guestrooms also feature kitchens, complimentary wireless Internet access, and microwaves.
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text:
Have you ever asked yourself, "Who would I like to become?" What people or events in life have hindered your reaching that goal? To achieve something, you have to remove all distractions that are blocking you from making progress. Fear stops most people from getting what they want — fear of failure, fear of success, fear of rejection, of the unknown, of disappointment. Then, of course, the "what if" fears always fuel these hindrances.
These opinions are not the Courier's. Callers are not required to identify themselves, so we can neither verify sources nor their motives. Call 876-3733.
One Friday evening in May 1976, an incident caused a lot of commotion on Flowers Road, which is off George Swindell Road and Highway 196, in Long County. The ordeal was something that no family ever wants to go through.
Relay for Life of Liberty County aims to engage community members and raise more money ahead of the May 15 event by participating in a challenge issued by The American Cancer Society.
Join other NFL fans to watch the Seahawks and Patriots battle in Super Bowl XLIX Sunday at Thunder Run in Club Stewart. Doors open at 5:30 p.m. Admission is free.
Community health-care facilities are expanding, and services are improving for civilians, veterans and military families. Medical facilities on and off post have seen growth in the past year, and more improvements are planned for this year.
American Red Cross supporters have a new way to help save lives in January during National Blood Donor Month through blood and platelet donation with SleevesUp virtual blood drives.
JESUP — Residents in Jesup and the surrounding communities now have one less reason to travel elsewhere for health care thanks to Wayne Memorial Hospital's addition of full-time cardiologist Dr. Gary Stern and an additional diagnostic tool, the cardiac CT.
SAVANNAH — A renowned palliative-care expert from Johns Hopkins is coming to Savannah at the end of the month as part of Hospice Savannah Inc. and The Steward Center for Palliative Care's seventh annual lecture series.
BRUNSWICK — What does it really imply if products are listed as all-natural or gluten-free? Which part of the label should consumers review first? What is the appropriate amount of beneficial nutrients?


text:
Telephone & Addresses
Lynn Gosling
11790 Stony Brook NY
Lynn Gosling
95442 Glen Ellen CA
Lynn A Gosling (Age: 72)
37923 Knoxville TN
Lynn Gosling, 61, Arlington, US, Henderson Dr, Apt 828
View Lynn's social profiles and photos on Facebook, MySpace, and +40 Networks.
Yellow Page contacts
Network Profiles
MySpace Profile: Neassa Zuchowski (baby girl) on Myspace
c) Nicki Lynn Gosling d) Nicki Marie Gosling e) Nicki Grace Gosling Please take the entire quiz (5 questions), or you can Create your own Quiz! plz try to
Twitter Profile: kali lynn gosling (@HelloKitty_Elmo) | Twitter
oct i live i newton fall ohio i have two brothers one is 7 and another is 5 i have 3 ...
Lynn Gosling – Google+
Lynn Gosling hat auf dieser Seite noch nichts mit Ihnen geteilt.
Real Estate Transfers 12/27/09 - News Sentinel Story
David Butler to William Gosling and Lynn Gosling, in Greywood Crossing subdivision, $206,500. Smithbilt LLC to Regina Osborn, in Millers ...
Management & Stakeholders
LYNN GOSLING of TN8 6HP - contact details and financial details of G & W PROJECTS (SOUTHERN) LIMITED and other associated companies.
Business Profiles
LinkedIn: Lynn Gosling | LinkedIn
View Lynn Gosling's professional profile on LinkedIn. LinkedIn is the world's largest business network, helping professionals like Lynn Gosling discover...
Lynn Gosling | California, United States | Birth Record | 563229718
Lynn Gosling is a person located in California, United States. January 08, 1908
classmates: Lynn Bennett-Gosling (Bennett)
Cornell University - Human Ecology, Ithaca, NY, 1999-2002
Bad news
findagrave: Gosling, Lynn V.
01/8/1908 - 05/26/1996, Stirling City (Butte County, California)
Hathaway Family Funeral Home Obituary: Printer Friendly Version
Camara, Lisa Gosling, Lynn Gosling, Thomas and Jamie(Mac ...
LYNN GOSLING (1908-1996) | Social Security Death Index
Lynn Gosling was born in 1908 and passed away in 1996 . Access the Social Security Death Index on Mocavo and find your ancestors for free!
Alle Ergebnisse für Mary Hungate in Alle Sammlungen - MyHeritage
Lynn Gosling Jack Deb Grobe Deborah Grosz Ellen Bob Hamilton Judy Heatherington John Cindy Heisler Glen Ann Hildebrand Kathryn ...
Ancestors of Margaret Ward Bromley:Information about Ashley Nicole...
Ashley Nicole Duvall (daughter of James Arnold Duvall and Vickie Lynn Gosling). Home | Help | About Us | Biography.com | HistoryChannel.com | Site Index | Terms of Service |
Obituary Guestbook | Kenneth Lucky Baldwin of Wheaton, Illinois |...
View Guestbook For Kenneth Lucky Baldwin of Wheaton, Illinois. Please join us in Loving, Sharing and Memorializing Kenneth Lucky Baldwin on this permanent...
Help Us Find a Cure
Bill & Lynn Gosling 5/4/2007, $25.00. Amy Goeb Good luck on your walk! 5/4/ 2007, $25.00. Karen Brand Jan, I'm happy to help! 5/4/2007, $25.00. Carolyn You ...
Andrew Gosling is fundraising for Youth Sport Trust
JustGiving - the easiest way to fundraise and donate to charity online
Related Documents
Lynn Gosling, Tunbridge Wells, TN8 6HP | Creditgate
Director search available online for Lynn Gosling, Tunbridge Wells, South East, TN8 6HP.
7th Grade
This way I can send you messages and reminders throughout the year. Sincerely,. Lynn Gosling and the Seventh Grade Teachers. Middle School Counselor.
Video & Audio
Lynn Gosling - YouTube
next; Play now · CIMG7101. by Lynn Gosling; 1 month ago; No views.
Mandi Lynn Gosling on Vimeo
Mandi Lynn Gosling is a member of Vimeo, the home for high quality videos and the people who love them.
Keeping in Touch - Morning Call
TWO MORAVIAN COLLEGESTUDENTS have been named the recipients of Fulbright fellowships, both to be used in Germany.Julie E. Anderson of Bethlehem, a double major...
Jubilee Furniture: Greetings from Jubilee Furniture!
09.04.2010 Congressman Peter Roskam has a beautiful Jubilee Furniture coffee table in his office in Washington DC and Bill and Lynn Gosling have purchased quite a bit of furniture from us for their new home in North Carolina (there ...
Andy's Fan Page: Liberty High School graduation features 620 ...
06.06.2011 Carlos Gonzalez Corina Lee Gonzalez Nicholes Richie Gonzalez Ricardo Eric Gonzalez Ruben Gonzalez Staphany A. Gonzalez Sara Nicole Gori Ava Lynn Gosling-Prutko Latifah Diana Graham Hannah Christine Graser ...
In Partnership With God: November 2010
John Engstrom, the athletic director at Stony Brook Academy knew Bill and Lynn Gosling. Bill was in capital development at Stony Brook.
Reports & Statements
pinterest.com: Lynn Gosling on Pinterest
Lynn Gosling is using Pinterest, an online pinboard to collect and share what inspires you.
Re: UK German family
Re: UK German family. Posted by: Matthew German, Date: January 21, 2001 at 11:55:47. In Reply to: Re: UK German family by Lynn Gosling, of 1170 ...
Harlings - Message Boards - AncestryLibrary.com
Lynn Gosling (View posts). Posted: 9 Feb 2005 1:54PM GMT. Classification: Query. Surnames: Harling. Sorry. Non of those names sound familiar. Our harlings ...
Paperless Wedding Australia: Traditions - The Wedding Cake
07.09.2010 Written by Lynn Gosling: Having now made several wedding cakes for friends and family I thought it would be interesting to look at how the custom of serving a special cake originated. In medieval times cakes made of wheat, ...
Barossa Personal Cooking Lessons
Cook up a Storm Individual cooking lessons are held weekly inthe Barossa Valley. For further informationcontact: Coordinator: Lynn Gosling
Amanda Lynn Gosling, 1295979334 NPI number, (203) 688-2259
AMANDA LYNN GOSLING. Get details of Amanda Lynn Gosling including phone number (203) 688-2259, npi number 1295979334, business and practice ...
Amanda Lynn Gosling, MD - Pediatrics, Wyomissing, PA, US
Amanda Lynn Gosling, MD - Wyomissing, PA, US - Pediatrics. Register to update your online profile.
Lynn Gosling | Meaning Pronunciation Origin of Baby Name Lynn Gosling...
Lynn Gosling : Baby Name Lynn Gosling Meaning,Pronunciation,Origin,Religion,Pronounce of Baby Name Lynn Gosling. Similar Names ,All about the name Lynn Gosling...
[il-talk] Lynn Gosling Update
Hi, Everyone, Lynn Gosling has asked me to let you know that she had back surgery last Monday morning at Edwards Hospital in Naperville.
About Amanda L Gosling - Kids Health Reference
female pediatric general/family doctor currently practicing at Reading Berks
Physical ...
Jamie Lynn Gosling Address & Voting Record of Newton Falls, OH |...
Jamie Lynn Gosling's Address & Voting Record of Newton Falls, OH. Get more information on Jamie. Voter id:OH0015814371
Lynn Gosling 1's profile - Hear the world's sounds
Explore the largest community of artists, bands, podcasters and creators of music
NPI 1295979334 Amanda Lynn Gosling in Wyomissing - Address, Taxonomy,...
NPI 1295979334 Amanda Lynn Gosling in Wyomissing - Practice Location Address, Taxonomy and Contact
Lynn Gosling corporate information
Lynn Gosling corporate information and associated companies
LYNN GOSLING director information. Free director information.
LYNN GOSLING - a free Director Summary including all company appointments. Instant Director Report for Risk Scores, County Court Judgements, Mortgages and...
Lynn Gosling's £5 contribution to Help the FoxG1 Foundation Start Our...
luck with the fundraising , such a good cause. Like 1 Comment ...
Albion Stitch: Albion Bezel inspiration
when one of my lovely students, Lynn Gosling, brought this ...
Southport Flower Show: Bee's Knees Garden & Birkdale Youth |...
Tweet Video x2: From the substantial resources of Sefton's own parks and gardens department, to the infectious enthusiasm of some young first-time gardeners...
2009 Results « Adirondack Hiking, Climbing, Skiing, Books | The...
Purveyors of fine Outdoor Equipment since 1975
0 200 134 20fde402e427dec3 232c9fc49b 3 0xffffff 0xffffcc 0xffffff ...
Dear Family in Ohio and Florida, Helen's father ("Doc") and my grandfather ("Daddy Joe") were brothers, thus Helen and my father (Lynn Gosling d.1996) were ...
Archive for November, 2010 | The Sundial | Page 9
Colors of a Soprano – Maria Elena Altany from Mandi Lynn Gosling on Vimeo. Published on November 16, 2010 By Julio Alberto Cruz ...
Running for the fun of it - AdirondackDailyEnterprise.com | News,...
SARANAC LAKE — More than 100 runners turned out this week for the Saranac Lake Fun Run.
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