In order to identify novel mutations affecting mechanosensation, we designed an assay that measures the behavioral response to tactile stimulation of fly bristles in mutant clones generated by the MARCM method. The combination of techniques allows for the identification of mechanosensitive mutations that would otherwise be lethal.
Murphy, T. P., Luu, D. D., DeSimone, J. A., O'Brien, T. C., Lally, C. J., Lindblad, J. J., Webster, S. M. A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster. J. Vis. Exp. (106), e53537, doi:10.3791/53537 (2015).
Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.
Humans rely on the ability to convert mechanical stimuli from their environment, such as touch, pressure, vibration, or sound waves, into sensory information that can be processed by the nervous system, in a process termed mechanotransduction. Many of the overall mechanistic features of mechanotransduction between humans and invertebrates are the same1, making Drosophila a useful model to study the molecular mechanisms of mechanotransduction. Drosophila melanogaster contain two sets of specialized sensory organs (Type I and II) that are capable of converting mechanical stimuli into action potentials. Type I mechanoreceptors have a neuron with a single dendrite or sensory process, surrounded by three support cells2. The Type I mechanoreceptors include bristle mechanoreceptors, hearing sensitive chordotonal organs (the Johnston's organ), and campaniform sensilla that convey information about wing beats3. The bristles that cover the dorsal side of the fly are the most abundant and easily accessible of the Type I organs. Similar to the extracellular environment of hair cells associated with hearing and balance in vertebrates, the support cells surrounding fly mechanosensitive neurons secrete a high potassium endolymph that creates an unusual concentration gradient for potassium1. Mechanosensitive neurons, of both the mammalian and fly systems, utilize this high extracellular potassium to depolarize the cell. In response to mechanical stimulation of the macrochaete bristles towards the body wall, the sensory neurons respond with a burst of action potentials driven by this potassium depolarization of the cell membrane4. The Drosophila sensory neurons that innervate bristles resemble the mechanosensitive cells of other organisms, including vertebrate hair cells, in both structure and function1,5. The accessibility of Drosophila external sense organs to experimental manipulation and the abundance of genetic techniques available to researchers make Drosophila an excellent model system to investigate the molecular underpinnings of mechanosensation.
In the fly, stimulation of a single sensory neuron that innervates a bristle leads to an observable behavioral response. Stimulation of different bristles evokes specific, reproducible behavioral responses, depending on the bristle that is stimulated. Upon tactile stimulation, decapitated wild-type flies exhibit a complex grooming reflex wherein they clean the area near the stimulated bristle with a patterned set of leg movements5-7. When homozygous for known single-gene mutations that interfere with learning7 or coordination and locomotor activity5, flies respond abnormally to mechanical stimulation. This grooming reflex is therefore a useful tool to study the effects of single-gene mutations on a specific, replicable behavior.
The robust behavioral response to stimulation of a single macrochaete bristle holds the potential to assist in identifying new genes involved in mechanotransduction. This protocol being used to test a collection of mutant flies for the absence of a behavioral response to indicate that the mutation interferes with mechanosensation. In the mutant collection selected for screening, the mutations cause lethality before adulthood, and therefore would be impossible to test using traditional adult behavior screens. Originally, this collection of lethal p-elements was combined with FRT recombination sites to test cell growth defects in clones. The clones were made specifically in the eye because adult flies can survive in the lab setting without functional vision8. However, removal of all mechanosensation can cause adults to be severely uncoordinated or die before eclosion5. This protocol uses a mosaic approach to circumvent the lethality of the mutations and allow for adult stage testing. A genetic technique called Mosaic Analysis with a Repressible Cell Marker (MARCM)9 is used to generate homozygous mutant cells in a limited number of adult fly sensory organs, while the rest of the organism remains heterozygous. These MARCM flies readily survive until adulthood, yet the bristles are homozygous for the lethal genes of interest.
MARCM allows for regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP), while the rest of the organism remains heterozygous at that particular locus and unmarked9. MARCM combines individual p-element mutations with five common genetic elements: GFP under the control of an upstream-activating sequence (UAS-GFP), Gal80 repressor protein under control of a general promoter (tub-Gal80), Gal4 transcription factor under control of a general promoter (tub-Gal4), the FRT recombinase enzyme expressed through a heat-shock controlled promoter, and a FRT recombination site10. By driving mitotic recombination through a heat-shock activated recombinase, a limited number of cells are made homozygous for the mutation and marked with GFP. GFP expression is repressed in heterozygous cells by the presence of Gal80 on the wild-type copy of the chromosome.
A heat-shock protocol for MARCM was optimized to induce recombination in the fly bristle external sense organs, while much of the organism remains heterozygous, and thus unmarked with GFP. Mosaic flies generated using this protocol contained homozygous mutations most frequently in the post alar or dorsal central bristles on the surface of the notum.
We have tested the utility of this combination of MARCM and the grooming behavior screen with a known mechanosensitive mutant, NOMPC. The ion channel, no mechanoreceptor potential C (NOMPC), is an essential component of the mechanotransduction pathway in Drosophila5,11-13. NOMPC belongs to the transient receptor potential (TRP) superfamily of cation channels5 and satisfies all of the criteria to qualify as a mechanosensitive channel in Drosophila14,15: 1) NOMPC is expressed in the ciliate tips of type 1 sensory neurons of Drosophila 13,16-18, 2) NOMPC null larvae do not have an electrical response to tactile stimulation13, 3) Ectopic expression of NOMPC in touch insensitive cells can induce sensitivity to mechanical stimulation13, 4) heterologous expression of NOMPC in Schneider 2 cells yields a mechanosensitive channel13, and 5) NOMPC adult mutants display defects in their response to mechanical stimulation5. Given this evidence, we predicted that NOMPC mutant clones would show an altered or inhibited grooming response in response to mechanical stimulation of bristles.
A MARCM-stock containing the NOMPC mutation was developed for use in a proof of principle experiment of the grooming assay. Mosaic flies were stimulated at macrochaete bristles containing homozygous NOMPC mutant cells. We expected an inhibition of the grooming response following stimulation of the macrochaete bristle. We found that only 2 of 14 mutant bristle flies tested gave a single response to repeated stimulation; most did not respond to stimulation of the homozygous mutant bristle. Having confirmed that this MARCM-based behavioral assay produces an abnormal grooming reflex in a known mechanosensitive mosaic mutant, this technique can be used in a screen for additional mechanosensitive mutations.
Maintain flies on cornmeal molasses media (6.5 g/L agar; 23.5 g/L yeast; 60 g/L cornmeal; 60 ml/L molasses; 4 ml/L acid mix; 0.13% Tegosept) or standard Drosophila media at 22 degrees Celsius in a 12-hr light/dark cycle.
Note: A Lower than standard temperature is used to keep MARCM-stocks healthy. This extends the developmental time between egg and adulthood to 14 days.
Use virgin females (approximately 1 - 8 days old) from a MARCM-ready stock containing the necessary genetic elements (GFP under the control of an upstream-activating sequence (UAS-GFP), the Gal80 repressor protein, the Gal4 transcription factor, a heat-shock driven recombinase, and a FRT recombination site).
Note: Our MARCM ready-stocks contain the following genotypes: elavGal4, UAS-CD8-GFP, hs-flipase; FRT40A, tubGal80/CyO; tubGal4/TM3Sb.
Note: The specific FRT site will depend on the location of the mutation in the genome. For example, FRT40A is used to test mutations on the left arm of chromosome 2, such as NOMPC (see10 for more details on MARCM protocols).
Cross MARCM-ready virgin females to males that contain a mutation of interest and its corresponding FRT site (for example, w-; NOMPC3, FRT40A/CyO; +/+). To control for effective MARCM induction, set a second cross using male flies containing the same FRT site, but without a mutation. Use an approximate ratio of 5 females: 1 male in crosses.
After allowing flies to mate for at least one night, move adults into new vials with fresh media. Allow flies to lay eggs in a dark environment for approximately 4 hr before again transferring the crosses into new vials. Note the start and end times of the egg laying period.
Note: Shorter intervals are acceptable when there appears to be a high number of eggs on the media before the end of the 4-hr interval.
Keep vials with a sufficient number of eggs (approximately greater than 20) and store in a 12-hr light/dark cycle at 22 degrees Celsius to be heat shocked later.
Note: Vials with a low number of eggs on the media (less than approximately 20) are unlikely to yield sufficient numbers of adult flies for behavioral testing and in the interest of efficiency it is recommended they not be retained for further testing.
Note: Our flies are raised with the lights on from 8 AM to 8 PM. We have found the evening time points lead to higher egg densities, therefore usually perform timed crosses between 4 - 8 PM and 8 PM-midnight. The flies lay their eggs at RT in a dark cabinet and then are returned to the incubator the following day so as not to disturb the regular light-dark cycle in the incubator.
Approximately 85 - 100 hr after egg laying, place vials into a water bath at 37 degrees Celsius for 1 hr. Ensure that the water level reaches above the height of the media in the vials, but does not fully submerge the vials to prevent the larvae from drowning.
Remove vials from water bath and allow a 1 hr recovery period at RT.
Place vials back into 37 degrees Celsius water bath for 1 hr.
Remove vials from water bath and store in incubator on 12-hr light/dark cycle at 22 degrees Celsius until eclosion.
Following eclosion, anesthetize flies on ice. Select flies with genotypes that contain all the genetic elements for MARCM and indicate GFP expression-marked clones will likely be observed based on the observable phenotype markers used in the parent stocks. Decapitate flies with iridectomy scissors.
Note: For example, we selected for flies without the genetic markers CyO (curly wings) and Sb (stubble bristle) to identify flies that would contain all the genetic elements necessary to create mosaic regions marked with GFP.
Note: Decapitation is necessary to prevent the adults from flying away when stimulated.
Place headless flies in a closed, moist environment and allow approximately 10 - 20 hr of recovery. Only use flies that right themselves when perturbed for further testing. Test flies within 30 hr of decapitation.
By observing decapitated flies under a fluorescence dissecting microscope, identify homozygous clones marked with GFP at the bristle external sensory organs. Record the bristle name and left or right side. To eliminate any potential bias, the lab member performing the bristle stimulation should be blind to the genotype of the bristle, wild-type or mutant.
Elicit a grooming reflex by deflecting the GFP marked bristle towards the fly body with a stiff hair or fine forceps and observe the leg response. Give a score of one to flies that lift their leg in response to bristle stimulation and a score of zero to flies that do not move their legs.
Note: Previous studies6,7 have characterized the leg response that is elicited by stimulating a particular bristle. This includes the leg that responds to each bristle and the proportion of flies that respond to stimulation of that particular bristle. We only scored for movements of the leg that we expected to respond based on Vandervorst's classification6.
Conduct five trials spaced 2 min apart.
The success of this protocol largely depends on the overall yield of testable flies, the effectiveness of the heat shock to induce mitotic recombination, and the ability to obtain a robust behavioral response in flies stimulated at 2 min intervals. Given that an estimated 12.5% of embryos contain the necessary MARCM elements and an estimated 16.67% of adults that eclose have the potential for GFP expression, it is critical to obtain a high number of eggs during timed lays. An example of a successful timed lay over a 4 hr interval that would be used in the rest of the protocol is shown in Figure 1A.
When the heat shock is performed 85 - 100 hr after egg-laying, the yield of GFP labelled external sense organs is highest, with most flies having one of the macrochaetae labelled. Numerous areas of interest may contain homozygous cells at the bases of testable bristles when heat shocks are performed within this time period, with the dorsal central and post-alar bristle being found labelled most frequently (Figure 2A). Time points outside of this window result in patches of GFP expression outside of the body regions of interest or no GFP expression (Figure 2B).
Flies naturally respond to a looming stimulus by initiating an escape response19. In order to prevent flies from escaping, we decapitated flies. The grooming response circuit remains intact in the ventral nerve cord5. Flies do not recover well if decapitated under carbon dioxide anesthesia, but recover in a few hours when decapitated with cold anesthesia on ice. After decapitation, flies should be stored in an enclosed, moist environment during the recovery period to prevent desiccation. While some flies may desiccate or be unable to stand (Figure 1B), only those able to stand (Figure 1C) are used for testing. Prior to testing, flies are checked for general responsiveness by watching for movement after a touch to the notum. Many flies remain responsive for several days, but testing should be completed by 24 - 36 hr after decapitation. Fly hyperactivity can be a problem during behavioral testing because an observer may have difficulty distinguishing between a true stimulus-dependent grooming response and general movements of the fly. In animals with excess movement not elicited by tactile stimulation, more reliable results are achieved by waiting for hyperactive flies to become quiescent.
We found that 47% of wild-type flies respond at least once when touched on a bristle five times, spaced at 2 min intervals to prevent habituation. Several groups have shown that stimulation of different bristles elicits a patterned response from a specific leg 6,7. Of the bristles that do elicit a grooming reflex following stimulation, the percentage of flies responding ranges from 38% (anterior scutellar) to 94% (anterior noto-pleural)6. In our experiments, wild-type control flies responded to a light touch from 14% (dorsal central) to 86% (noto-pleural) of trials (Figure 3). In a MARCM-based screen with unknown mutants, those flies containing a mutation not involved in mechanosensation can be expected to exhibit a reliable behavioral response similar to that observed in wild-type control animals.
The known mechanosensitive mutant, NOMPC disrupted the grooming response as expected. Control flies responded to bristle stimulations at homozygous cloned post-alar bristles 37% of the time (Figure 4). In contrast, animals with bristles that were homozygous for NOMPC responded 2% of the time (Figure 4). While wild-type flies respond differently to stimulation of different bristles, these results suggest that a mechanosensitive mutation, such as NOMPC, nearly eliminates the grooming response. In a screen with unknown mutants, several mutations involved in mechanosensation that inhibit the grooming response are expected.
Figure 1. Proper Rearing Conditions are required for Optimal Behavioral Testing. (A) A sufficient number of embryos are needed following the 2 - 4 hr egg laying period. The number of embryos after a 4 hr timed lay is likely to yield a sufficient number of adult flies for behavioral testing. (B) During the recovery period following decapitation, some flies do not survive and cannot be used in further testing. (C) A decapitated fly that was properly stored in a moist environment during the recovery period is testable in a behavioral assay. Please click here to view a larger version of this figure.
Figure 2. MARCM Clones in Bristles when Heat-shocked between 85 - 100 Hr After Egg-laying. (A) GFP expression in adult flies when heat shocked 98 - 101 hr after egg laying. As many as 10 macrochaete bristles contain homozygous mutant external sense organs and are appropriate for testing in the grooming response behavioral assay. (B) GFP expression in adult flies when heat shocked 161 - 164 hr after egg laying. Note the lack of GFP expression at the bases of the macrochaete bristles, indicating no patches of homozygous cells at a testable external sensory organ. Please click here to view a larger version of this figure.
Figure 3. The Frequency of Response to Tactile Stimulation is Dependent on Bristle Identity. The total number of responses of an individual fly to stimulation of a single bristle was divided by the number of stimuli to give a percent response. Percentages were averaged for each bristle (N = # of flies tested, 22 Post Alar, 21 Dorsal Central 14, Scutellar, 7 Notopleural). Please click here to view a larger version of this figure.
Figure 4. NOMPC Mutants Do Not Respond to Tactile Stimulation. Stimulation of the post-alar bristles elicits a response in wild-type MARCM clones. However, stimulation of MARCM clones on the post alar bristles that were NOMPC3 -/- (green bar) resulted in few responses (N = 23 wild-type, and 12 NOMPC -/-). Please click here to view a larger version of this figure.
This protocol utilizes an adult behavioral assay to screen for mutations that affect mechanosensation in Drosophila. Because the collection of mutants contains lethal p-element mutations that would preclude screening as adults, this protocol makes use of a complex genetic technique first described by Lee and Luo, (1999) and detailed as a protocol by Wu and Luo, (2006) to circumvent adult lethality. MARCM induces mitotic recombination between homologous chromosomes to generate clonal regions of homozygous cells. These regions are marked with GFP because of the loss of the repressor protein, tub-GAL80 that occurs during mitotic recombination. By using MARCM to generate cells which are homozygous at a particular genetic locus, gene function can be studied in adult flies using a variety of methods, including the behavioral assay described in this paper.
The primary factor limiting both the number and the specific genes that can be tested is the presence of a FRT site within the genome of the fly. Fly stocks must contain a FRT site on the chromosomal arm of the mutation of interest. MARCM ready fly stocks must contain an identical FRT site as the mutant chromosome to allow for mitotic recombination. In our lab, we developed fly stocks containing MARCM backgrounds with FRT40A and FRT42D sites. These stocks will allow us to screen any adult lethal p-element already combined with a corresponding FRT site on the 2L and 2R chromosome arms, respectively. To maximize the potential of MARCM in screens for mutations that affect mechanotransduction or the grooming reflex, additional fly stocks with MARCM backgrounds capable of testing genes on both arms of the third chromosome and the X chromosome will be created. Once developed, these stocks will allow for most adult lethal p-element mutations combined with a FRT site to be tested for involvement in mechanosensation.
The timing of the heat shock protocol has been developed to maximize the number of homozygous cell regions at the external sensory organs, while leaving much of the rest of the fly heterozygous at the locus of interest. Numerous areas of interest can be marked by GFP expression following a double heat shock, allowing for multiple bristles to be tested (Figure 2). The MARCM protocol provides an excellent opportunity to create homozygous lethal mutations in specific regions of interest, but our MARCM protocol relies on the heat shock induction of recombinase activity, making it a random process as to which bristles will ultimately be testable. Because each bristle elicits a specific leg grooming reflex6,7, it was important for us to confirm that the responses in wild-type MARCM clones are comparable to those previously described. Maximizing the efficiency of a MARCM-based screen requires the ability to test any of the macrochaete bristles for a mutant response and to compare these data against the response rate from a corresponding bristle in a MARCM-ready stock. We have confirmed the results of Corfas and Dudai7 in bristles expressing GFP in the MARCM flies.
The ability to readily distinguish between normal and abnormal fly behavioral responses is critical to determine which mutations interfere with mechanotransduction. A mutation in a gene involved in mechanotransduction or other aspect of the grooming reflex should inhibit the leg response; a significantly lower response rate at the corresponding bristle in a MARCM-ready stock is expected. The behavioral response rates in mosaic NOMPC and wild-type flies serve as a proof of the principle that the MARCM-based behavioral screen can be used to differentiate between mechanosensory mutants and normal flies. We have shown that in NOMPC mutant mosaic clones, the grooming reflex in response to bristle stimulation is nearly abolished. Only two flies of the fourteen tested exhibited a leg response. Each of these flies responded only once over the course of testing, suggesting that a MARCM-based behavioral assay is effective at identifying a mechanosensation defect. An observer can distinguish between mechanosensitive mutant flies and flies without a mutation or with a mutation that is not involved in the grooming response. Because of the high degree of variability in wild-type responses, the screen will identify strong alleles, but will likely miss more subtle changes in behavior due to weaker alleles. The collection of mutants to be screened includes only homozygous lethal alleles, so it is likely that these are strong or null alleles and are likely to abolish protein function. However, the screen would miss alleles that may be lethal in development, but produce a more subtle mechanosensory defect when made homozygous in a more limited manner by MARCM.
We chose an all-or-none scoring system for simplicity. This method yields a generous score compared to the system used by Corfas and Dudai7 that assigns a score of 0 - 4 depending on the extent of the leg movement. In the all-or-none system, any response is given full credit with the assumption that any response to stimulation indicates that the sensory system is detecting the stimulus and signaling to the motor system.
This screen may identify mutations that affect any step in the behavioral circuit. Further testing, such as electrophysiology (as in Kernan et al5) or calcium imaging, would have to be used to determine whether the mutation directly affected mechanotransduction or some other aspect of the development or function of the sensory-motor reflex arc.
Drosophila melanogaster provide an excellent model system for the study of mechanosensation. The external sensory organs share many structural and functional similarities with the hair cells of the mammalian inner ear20. Elucidation of the molecules involved in this process in Drosophila has the potential to reveal mammalian homologs and orthologs. Screens aimed at discovering mutations that disrupt mechanosensation will pave the way for subsequent cell biological, electrophysiological, and biochemical studies to further identify and characterize these molecules, deepening our understanding of this process in both flies and mammals. A MARCM-based screen for mutations affecting mechanosensation will use a well-established genetic technique paired with a grooming response behavioral assay to reveal new genes involved in mechanotransduction and the fly grooming reflex.
The authors would like thank the Bloomington stock center, Liqun Luo, Charles Zuker, and Lily and Yuh Nung Jan for the generous sharing of fly stocks and the following for funding: SOMAS-URM (to JD and SW), Bachelor Ford Faculty Summer Fellowship (to SW), BD Corporation Summer Research Fellowship (to CL and DL), The Renee and Anthony M. Marlon, M.D. '63 Summer Research Fellowship (to DL)James C. '75 and Jane Colihan Summer Research Fellowship (TO) through the Alumni/Parent Summer Research Fund of the College of the Holy Cross and the Stransky Foundation Summer Research Fellowship (to TO). Special thanks to the Department of Biology and the Dean's Office at College of the Holy Cross for the support of all the work in the lab.
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The court of appeals today filed a case which is recommended for publication. The case is Glidewell v. Glidewell, 2014AP1957. While the case seems appropriately decided, one has to wonder why it is recommended for publication as it does not address an important issue: The effect of DV on custody.
¶4…waived her right to raise the domestic violence presumption set forth in Wis. Stat. § 767.41(2)(d) in this post-judgment motion because the domestic violence she relies on occurred prior to her original divorce judgment, at which time she stipulated to joint custody. Jill cannot now assert the domestic violence presumption based on the pre-divorce incident and without asserting any post-judgment domestic violence facts.
This does not seem particularly revolutionary. In addition, it's difficult to believe that this change-of-mind occurs frequently enough to warrant a published opinion.
The opinion also affirms the order of Milwaukee Country Circuit Judge Fredrick Rosa in continuing joint legal custody, but awarding Jill final decision-making authority in the event of a dispute regarding medical care for the children and Herbert final decision-making authority in the event of a dispute regarding educational decisions for the children. The appellate court likewise affirmed this part of the order as a reasonable exercise of discretion.
The issue which both the trial court and appellate court avoided by ruling that Jill had waived the DV presumption against joint custody is whether there is a conflict between this presumption and the best interests of children. Put another way, can a man (yes, DV can and does occur the other way around, but it's far more common with the male being the perpetrator) be a DV perpetrator but still be an appropriate decision maker for minor children.
To quote from The Wizard of OZ: "I'm a very good man. I'm just a very bad Wizard." So, is it possible to be a very bad husband (and maybe even a very bad person) but still be a good (notice that I removed the modified "very") father?
Discuss amongst yourselves. This case does not answer that question.


A tight contest is expected when Girona square off against Getafe in a Spanish La Liga fixture at the Estadi Municipal de Montilivi. The match is predicted to go under 2.5 goals at odds of 1.48 while both teams are likely to settle for a draw.
Girona have been blighted by inconsistency all season having claimed five wins and suffered six defeats in their 15 matches. They have earned just two victories in their last five fixtures and head into Friday's clash off the back of losses in their last two matches.
Eusebio Sacristan's men were beaten last time out in the La Liga by title chasers Sevilla. They offered very little threat against the Andalusians and could only muster two shots on target as they fell to a 2-0 defeat at the Estadio Ramon Sanchez Pizjuan.
Girona are heavily reliant on striker Cristhian Stuani, who has scored 11 of the team's 17 goals this season and is priced at 2.85 to score on Friday. The Uruguayan has failed to find the net in his last two matches which has coincided with back-to-back defeats for the Catalan club.
Getafe are among a clutch of teams currently chasing a Europa League place, although they are just three points above their hosts on the table. They are unbeaten in four of their last five matches with two wins in that run.
Azulones earned a 1-0 victory at home against Real Sociedad in their previous La Liga outing last Saturday. Jorge Molina struck the only goal for Jose Bordalas' men in the third minute as they held off some intense pressure from the visitors to secure the win.
The visitors have also found the net just 17 times this season with Molina the team's top scorer with five goals. The striker is likely to be partnered in attack by Angel Diaz, who has failed to score in his last 10 matches.
Girona have claimed three wins and suffered just one defeat in six previous matches against Getafe. They were unbeaten against the Madrid based club last season, earning 1-0 victory at home before settling for a 1-1 draw in the reverse fixture.
Stuani scored in both fixtures against Getafe last season and will once again be the one to watch for Girona. The striker often carries the bulk of the attacking threat for the team but could be a marked man on Friday.
Getafe have also had their problems in attack this season with Molina and Diaz combining for just eight goals. They should struggle against a Girona team that has conceded only three goals in their last four matches.
Girona have seen under 2.5 goals in 6 of their last 8 matches (La Liga).
Girona are unbeaten in 5 of their last 6 matches against Getafe (La Liga).
Getafe have seen under 2.5 goals in 9 of their last 10 away matches (La Liga).
Getafe are unbeaten in 9 of their last 10 away matches (La Liga).
Getafe have scored at least 1 goal in 9 of their last 10 away matches (La Liga).
The attacking problems for Girona and Getafe mean a number of their matches this season have gone under 2.5 goals. Friday's fixture is expected to follow a similar pattern with both evenly matched teams tipped to settle for a draw.


In late July, project co-director Gordon H. Chang presented two talks on the Chinese railroad workers to Stanford alumni and friends at the Stanford Sierra Camp on Fallen Leaf Lake, near Lake Tahoe. Two hundred audience members attended the talks, which focused on the work the Chinese completed in the Sierras. Many in the audience had seen the rail line in the area above Donner Lake and near Truckee, but didn't know its history.
Chang spoke about tunneling and constructing "China Walls," the massive retaining walls that hold up parts of the rail line. The director of the camp noted that he had heard that several of the stone walls around the camp had also been built by Chinese. History was right there!


text:
George Lucas breaks ground on massive Museum of Narrative Art
George Lucas is building a massive art narrative art museum in Los Angeles. Photo Credit: Flickr/Raytwist35
George Lucas is building an art museum that will house popular art and 'Star Wars'memorabilia
by Lawrence Lease (article) and Milos Kitanovic (video)
March 15, 2018 at 3:46 AM March 15, 2018 at 3:46 AM
George Lucas has broken ground on his $1.5 billion-dollar Museum of Narrative Art. George Lucas was part of a ground-breaking ceremony held the site of the future museum in Los Angeles' Exposition, which is expected to be completed in four years.
Lucas delivered a passionate speech before a crowd of elected officials and distinguished guests including Steven Spielberg. Lucas acknowledged that it is an art museum, but he wants people to see it as an anthropological museum. George Lucas said that popular art tells us what a society aspires to be and what they want and who they truly are. It's a narrative story of their lives and beliefs.
George Lucas' art museum to be built in Los Angeles
Last year legendary film director George Lucas announced that Los Angeles would be the location for the new museum. Lucas said that he chose Los Angeles and Exposition Park because of its relative closeness to his alma mater the University of Southern California and hundreds of local high schools. The property can also be easily accessed by public transportation.
And the shovels are in! George Lucas, Mellody Hobson, Mayor Eric Garcetti, and the entire Lucas Museum team have officially launched construction in Exposition Park. pic.twitter.com/sNla299ZLn
— Christi Carras (@christi_anne96) March 14, 2018
According to ABC 7, this new revolutionary museum will be 300,000 square feet and will display dozens of collections and pieces from famous artists including David Hockney and Norman Rockwell.
The museum will also display Star Wars memorabilia as well as illustrations from children's books. During the ceremony, George Lucas was able to expand on where he got the inspiration for the museum.
George Lucas said that narrative art has begun to be lost in the modern city and he wants to support all the art that everyone loves but doesn't see. New renderings of the futuristic-looking museum were released at the ceremony.
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The museum is being designed by popular Chinese architect Ma Yansong. The renderings gave a glimpse into the inside of the museum as well as the landscape.
Thrilled to join Mellody Hobson and George Lucas for the groundbreaking of the @LucasMuseum of Narrative Art, a $1 billion investment in our city that will inspire generations of Angelenos! pic.twitter.com/vzRYGchT2V
— Mayor Eric Garcetti (@MayorOfLA) March 14, 2018
Los Angeles welcomed the new museum
Lucas had also previously attempted to locate the museum in Chicago and San Francisco but both communities strongly opposed the plans.
Los Angeles Mayor Eric Garcetti was ready and willing to help make Lucas' dream a reality. Garcetti at the ceremony even welcomed Lucas into Los Angeles and said that "this is a city you belong in." The museum received an endowment of nearly $400 million, becoming one of the largest private gifts given to a city in history. Garcetti also thanked George Lucas for his vision, which he says will inspire visitors from all over the world. This museum will breathe new life into Exposition Park.
'90 Day Fiancé': Ariela Weinberg's latest weight loss pictures have stunned fans
Tom Brady keeps 'bromance' with Edelman alive, makes fun of failed 'high-five' with ref
Bruce Arians: 'I allow Brady to coach, I just sit back sometimes and watch'
Lawrence Lease
Lawrence M. Lease writes frequently on politics, entertainment and sports. He has been published on Communities Digital News, Eye on America News and in The People's Paper and his college paper the USF Oracle.
Follow lawrence on Facebook Follow lawrence on Twitter Follow lawrence on Linkedin
Read more on the same topic from Lawrence Lease:
'Rogue One' Series: Jimmy Smits potentially returning as Bail Organa Disney Theme Parks are expected to lose $21 billion during shutdown 'No Time to Die' could potentially get pushed back to 2021
Blasting News recommends '90 Day Fiancé': Molly Hopkins health in stable condition after alleged assault by Olivia '90 Day Fiancé': Yara's pregnancy result could be delayed longer '90 Day Fiancé': Rose gives bedroom tour, fans stunned to see her standard of living '90 Day Fiancé': Jenny opens up to TLC producer after learning about Sumit's arranged marriage 'Harry & Meghan: A Royal Romance' has an international cast 'Criminal Minds': The BAU team has taken the power back '90 Day Fiance: The Other Way': Kenneth says his relationship is not acted Video '90 Day Fiance: The Other Way:' Sumit is facing backlash for choosing Jenny Video

redpajama_set_name: RedPajamaCommonCrawl


The monitoring of winter road surface conditions (RSCs) is essential to transportation agencies and the traveling public, since the former needs to be aware of the location and severity of existing RSCs in order to effectively maintain safe roadways with minimal environmental impact, while the latter uses RSC information to make informed travel decisions. However, current RSC monitoring practice still relies on methods that are time-consuming, labour-intensive and lacking in objectivity, therefore limiting their ability to provide sufficient spatial and temporal coverage across a road network. This research was motivated by the need for accurate, timely and reliable RSC monitoring for winter maintenance personnel and the travelling public. To achieve this objective, the field performance of a smartphone-based RSC monitoring system was evaluated on a section of Highway 6 in Ontario, Canada during the winter of 2014. A comparison between this system and current monitoring methods indicated that the former was capable of providing reliable results particularly at the maintenance route level; however, classification accuracy was found to vary according to RSC type. To improve the results produced by the smartphone-based system, this thesis proposes a connected- vehicle (CV) based RSC monitoring system that utilizes Road Weather Information System (RWIS) data in addition to the smartphone-based system's data. Three techniques in artificial neural networks (ANNs), random trees (RTs), and random forests (RFs) were tested as the underlying models of the CV system, and the results indicated that all three models successfully increased the classification accuracy of the smartphone-based system. RFs were found to provide the most accurate RSC classifications for the standard (three-class) classification scheme while RTs were found to be most accurate when using a more detailed (five-class) classification scheme. Model transferability was also tested using data captured from a different test site during the winter of 2015; and it was found that although the proposed CV system significantly increased the reliability of RSC classifications, the underlying models were non-transferable and would therefore require local calibration before being used at different sites across a road network.


text:
The Real Life Kawaii Princess
By Shannon Lane @Shannonroselane
A LARGE, colourful bow in her pastel pink hair, doll-like makeup and a cutesy outfit topped off with a sprinkle of glitter is 32-year-old Michelle's every day style
Videographer / director: Marcus Hessenberg
Producer: Shannon Lane, Ruby Coote
Inspired by the Japanese culture derived from the district Harajuku, Michelle, from Portsmouth, says that the quirky trend has helped her gain confidence.
Michelle even extends her passion for Kawaii Japanese fashion to her bedroom, which is adorned with cute trinkets and countless stuffed toys.
She told BTV: "When I dress up, it's a little bit like a character. It makes you feel more confident and a bit more bold, and it's just nice to be different.
"There are lots of different Japanese fashions that I'm into. Now I've created my own style from lots of different inspirations.
"I would say I have been dressing like this for about six years.
"I first started to learn about Japanese fashion when I was at university, I studied Fashion and Textiles. A book with different pictures of the street-fashion from Harajuku really inspired me to start a project on that and that led me to develop my brand as well."
Michelle started her own business creating and selling clothing, jewellery and accessories inspired by Japanese fashion under the name Roxie Sweetheart.
"My style does reflect my personality, which is cute and fun and colourful," Michelle said.
"With clothes, I generally go with pastel colours. Occasionally I do wear black and other colours but mostly it's all like pastels, bright colours. Kind of like rainbow pastels. I really like anything iridescent or colour graphic, kind of sparkly.
"I think I probably own about 50 pair of shoes. They are obviously all pastel and all cute in some way."
Now, Michelle can hold her head high in her giant pastel bow, but she hasn't always been this confident.
She said: "Growing up, I didn't have that much confidence. It's hard for most teenagers to fit in and I didn't really know where I fitted in and I always had an interest for fashion but I didn't have the confidence to actually express that.
"I slowly got more and more into fashion but it was very gradual. So I would wear something and then I would wear a bit more, and a bit more and so every time I just build up a little more confidence and now I feel like I'm at a point where I could be who I am and do what I want.
"I don't feel awkward about it anymore. It's really nice because I think actually the fashion has enabled me to express myself and has given me that confidence.
"If someone says to me 'oh it's childish' I think maybe from an outsider's point of view or someone who doesn't know anything about the fashion, I can kind of see why someone might think that.
"But for me it's very much a creative thing. I'm such a creative person, that's what it is ultimately. I think when someone says that something's childish - but what makes it for a child? If it makes someone feel good about themselves then why does it need to be for a child?"
Michelle surrounds herself with everything pink, fluffy and sparkly - including in her bedroom and baby pink Fiat 500.
She said: "My favorite colour is pink obviously. In my room, everything is pink.
"My room is my place I go to relax and being surrounded by all these nice things, again it's more inspiration."
Michelle has not only found herself through her fashion, but also a close-knit group of friends who she met through Japanese style conventions, who are also into the quirky style.
Her friend Rosanna said: "The most common reaction from the outsider is 'Why are you wearing kid's stuff?'
"Kids do like it. I mean, in Japan it's for people with expandable income. So, it is people in their late teens and adults and all the way up to even old age in Japan."
Michelle added: "If it makes you feel nice it makes you feel happy - I think in this world it's important to have this escape, to have these things to make you feel good."
After coming out of her shell and gaining her confidence, Michelle's aim is to now inspire others to do the same.
She said: "My ultimate goal is to inspire people, even if it's just a little bit. That's my main aim, I just want to inspire other people to express themselves and to get into the fashion and to be not be afraid to do what they want and be what they want."


South Dakotans are headed to the polls to select a GOP governor and House candidate. See results here tonight.
Kristi Noem (gubernatorial) and Dusty Johnson (House) both secured convincing wins on Tuesday night.
Polls have closed in Sioux Falls and other locations in Eastern South Dakota, but don't expect results anytime soon.
Sixteen polling locations in Pennington County and three in Hughes County will stay open late Tuesday to accommodate voters.
The Secretary of State's office Tuesday afternoon announced that the locations would remain open for an extra 10 minutes or up to an extra hour and 45 minutes in some areas.
Voters across the state will have to wait until the final polls close at 9:45 p.m. Central / 8:45 p.m. Mountain to see voting returns.
The counties reported delays in getting voters into the polls earlier in the day when they're electronic poll book systems temporarily lost service.
Other counties moved to paper poll books to track voter registrations and provide appropriate ballots.
In a statement, the Secretary of State's office said it doesn't operate those electronic pollbooks. BPro, Inc. a Pierre-based company, operates the software.
Poll workers were grim as the early voters arrived to cast their ballots in the South Dakota election for June 5.
Even at Ronning Branch Library, where turnout has been in the hundreds by 9 a.m. in the city's recent mayoral elections, the mood was dour.
Looking at the 50 ballots cast by 8:05 a.m., one poll worker compared it to a school board election — not a glowing review, given that those elections struggle to break into the double-digits when looking at the percentage of people voting.
The voters who did come out, however, were generally excited to share their reasons for voting.
Tom Borchard said his vote for Congresswoman Kristi Noem to become South Dakota's governor stemmed from her opponent Attorney General Marty Jackley's treatment of the state's EB-5 scandal.
"It made him seem like a typical shady guy," Borchard said.
Craig Sletten said he also voted for Noem, and voted to send Dusty Johnson to Congress.
He'd met both of them, he said, and thought Johnson had the enthusiasm the state needed in Washington.


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Rams inability to execute with game and MW title hopes on the line, erases some of the good marks they received early.
CSU's football team passed the early tests Saturday night at Wyoming, scoring on its first possession, leading at halftime and by four points early in the fourth quarter.
The Rams were running the ball effectively on a snow-slicked field that made passing unusually difficult in the second half.
When it came time for the final exam, though, the Rams bombed.
They failed to convert a fourth-and-6 play from the Wyoming 35-yard line midway through the fourth quarter, then let the Cowboys march 69 yards in seven plays for the winning touchdown in a 16-13 victory.
That touchdown knocked Colorado State University out of contention for the Mountain West title and left the Rams 0-3 versus rivalry games with trophies going to the winners for the second straight season.
It would be easy to write off the loss to the weather, cold rain in the first half that turned to snow at halftime and blanketed the field. But, as CSU coach Mike Bobo was quick to point out afterward, both teams were playing in the same conditions.
Wyoming made the plays it needed down the stretch to win the game, CSU didn't.
So the Rams, who had so much to play for, get a "D" for this one.
Nick Stevens completed 8 of 14 passes for 110 yards in sloppy conditions. He failed to throw a touchdown pass for the first time in eight games. The offense scored one touchdown, settled for field goals on two other possessions and twice turned the ball over on downs in Wyoming territory.
Dalyn Dawkins ran for more than 100 yards for the fifth time in six games, gaining 154 yards on 29 carries while becoming the Rams' first 1,000-yard rusher in a season since Dee Hart in 2014. Izzy Matthews had 41 yards and a touchdown, and Rashaad Boddie gained 19 yards on four carries. CSU had 235 yards rushing. The Rams would have gotten an A here if not for their inability to finish drives with touchdowns and pick up a critical first down on fourth-and-6 late in the game.
It's hard to find too much fault with a unit that opened holes for a strong ground game. CSU averaged a respectable 4.3 yards a carry. But the Rams' front five allowed Wyoming to sack Stevens twice and hit him hard after a couple other throws in a game in which he only threw 14 passes. There also was a costly holding penalty that stalled a drive late in the second quarter.
CSU's defensive front played relatively well early on, limiting the Cowboys to just 102 rushing yards and 137 yards of offense in the first half. But they struggled to take down Wyoming quarterback Josh Allen when they had chances for sacks and in the fourth quarter. They also allowed the Wyoming quarterback to bull his way straight ahead for gains of 5 and 6 yards to set up the winning touchdown and a 2-yard run on third-and-1 with 2:25 remaining to seal the victory.
Josh Watson led the Rams with 11 stops, and Evan Colorito made a tackle in the backfield for a 7-yard loss on a third-down play that forced the Cowboys to settle for a field goal instead of touchdown after picking up a first down at the CSU 2-yard line. Max McDonald recovered a fumble in the fourth quarter, the game's only turnover. But the linebackers are the heart of the defense, and the defense failed the Rams when they needed it most, unable to stop straight-ahead runs on the drive that ended with the winning touchdown.
The Rams limited Allen to just 10 pass completions on 20 attempts and 138 yards through the air. But the passes they allowed included two for first downs on third-and-long plays to keep drives alive, one for a 21-yard gain to the CSU 2-yard line and another for a 17-yard gain on the opening play of the winning touchdown drive.
Although Wyatt Bryan made two field goals, and punter Ryan Stonehouse averaged 46.5 yards on four punts, a fumbled hold that prevented Bryan from attempting a 33-yard field goal late in the third quarter was a game-changing mistake.
It's easy to pile on the coach when things don't work out, and CSU fans were quick to do so on social media. But the decision to stop calling pass plays as the snow was piling up late in the game was understandable, given the troubles Stevens said he was having with his footing on his drop-backs and the inability of his receivers to make cuts.
CSU was leading and had the opportunity to put the game away without risking an interception. A punt from its own 19-yard line on fourth-and-7 with 2:45 remaining and two timeouts drew plenty of criticism. But it was the defense's inability to make a stop, allowing Wyoming to run for two first downs to run out the clock, that made it a bad call instead of the right call.
Ultimately, the Rams lost a game they not only needed to win, but in many respects should have won. Their inability to hold on to the lead and win a close game against their biggest rival and with the Bronze Boot and a shot at the Mountain West's Mountain Division title on the line suggests they haven't made as much progress this season as it appeared they had while they were winning their first four conference games.


Blackinton MIB1345 Mini Fire Maltese Cross Badge - MIB1345 - 1"
Home > Blackinton > Police Badges > Mini Police Badges > MIB1345 Mini Fire Maltese Cross Badge - MIB1345 - 1"
MIB1345 Mini Fire Maltese Cross Badge - MIB1345 - 1"
Keep your firefighting uniform looking fresh and unique with the Mini Fire Maltese Cross Badge from Blackinton. Featuring a classic Maltese style cross, this badge is a miniaturized version of the full-size favorite. Its center circle can be filled with virtually any seal the user prefers, making it perfect for firefighters of all levels, ranks and positions. Top and bottom bars are left blank to be engraved with custom lettering such as the wearer's name and unit number. The badges can be secured with a safety catch or used as a money or wallet clips. Because of their customizability, this badge makes a great gift for friends and family members.
©2002-2019 CopsPlus.com: Blackinton MIB1345 Mini Fire Maltese Cross Badge - MIB1345 - 1"


text:
Everything You Ever Wanted to Know About Angostura Bitters
There's a story behind that oversized label.
By Nate Erickson
The word "cocktail" is tossed around pretty loosely these days, used to describe just about any liquid sum that is greater than its parts. That wasn't always the case. When the term was first coined by a newspaper editor back in 1806, it referred specifically to a beverage comprised of spirits, sugar, water, and bitters. Today, we call it an Old Fashioned, and anyone who's ever made one, well, the old-fashioned way, knows it can't be done without Angostura bitters.
But what are Angostura bitters, and how did they earn their place in some of the most classic cocktails on the planet? What's with that label? In the interest of helping you go to the bar smarter, here are five facts worth learning about the best-known bottle of bitters.
Angostura was developed as a cure for upset stomachs.
As is typical in the spirits world, Angostura's place in society pre-dates its spot at the bar. All the way back in 1824, Dr. Johann Siegert—surgeon general for Venezuelan military leader Simón Bolívar—developed the stuff as a medicinal tonic for Bolívar's army. Created from a blend of herbs and spices, it was intended to cure upset stomachs. Originally called Dr. Siegert's Aromatic Bitters, it would later be renamed for the Venezuelan city of Angostura (now Ciudad Bolívar) where it was concocted. Operations eventually moved to Trinidad, where it is still produced today.
Angostura Bitters - 16 Ounce Bottle
That label was a mistake.
Besides its bright yellow cap, what really distinguishes a bottle of Angostura bitters is the label: It's too big. As legend goes, once Siegert's sons took over the business from their dad, they set out to market the bitters however they could, which included entering them in a competition. In a scramble to get their product ready for judging, one brother was assigned the task of retrieving bottles, while another went to print labels. Due to a miscommunication or mistake, they ended up with labels too big for their bottles, or bottles too small for their labels. By the time they realized the error, it was too late to correct. Though Angostura lost the competition, a friendly judge suggested the brothers make that label their signature. The advice stuck.
The recipe is a closely guarded secret.
Allegedly, only five people in the entire world know the exact combination of herbs and spices that go into a bottle of Angostura. As the only people with the recipe, they've even made a pact to never fly together or so much as eat together at the same restaurant, should, god forbid, the worst happen, according to VinePair. Guessing how much of this is fact, fiction, or just good marketing is all part of the fun, but it's clear the secret formula is kept close to the chest.
Some people use it as a cooking ingredient.
As anyone who has ever cracked an old cookbook knows, the sixties and seventies were a weird time for food. (Check out this Twitter account for examples.) In 1961, the brand capitalized on some of that weirdness, releasing The Secret of Good Taste: The Angostura Cookbook. From turkey to pies to bean soup, the book suggests a number of ways to incorporate bitters into everyday cooking. Why not?
Wisconsinites drink it by the glass.
There's a bar in the Badger State called Nelsen's Hall and Bitters Club that serves full shots of Angostura to patrons, a practice that's been kept alive since 1920. It began as a means of skirting the rules of Prohibition—it's simply a medicine for upset stomachs, after all—but as Punch reports, the tradition has lived on, giving Wisconsin the unique distinction of selling more Angostura per capita than any other state.
How to Make a Classic Old Fashioned
Nate Erickson Senior Lifestyle Editor Nate Erickson is the senior lifestyle editor for Esquire.com.
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text:
Q: Properties of oscillation Let $(X,\tau)$ be a topological space, $(M,\operatorname{d})$ a metric space and $f:X \to M$. Then

Definition:
The oscillation of $f$ at $x \in X$ is
$$ \omega_f(x) = \inf_{x \in U \in \tau} \operatorname{diam}(f[U])$$

There are two theorems that I am trying to prove. They are generalisations of some other theorems known to be true

*

*For any $\eta > 0$, $\{x\mid \omega_f(x) < \eta\} \in \tau$

*$f$ is continuous(open preimages for open sets) iff its oscillation is $0$.

It seems like that (1) could be proved using the fact that preimage preserves subsets, but I don't know how to show (2). Are they true or there are counterexamples?

A: For (1), if $\omega_f(x)<\eta$, then there is some open neighborhood $U$ of $x$ such that $f[U]$ has diameter $<\eta$.  For any $y\in U$, we then also have $\omega_f(y)<\eta$ because $U$ is also a neighborhood of $y$.  So $U$ is a neighborhood of $x$ contained in $\{x\mid\omega_f(x)<\eta\}$.  Since $x$ was arbitrary, this means $\{x\mid\omega_f(x)<\eta\}$ is open.
For (2), $f$ is continuous at $x$ iff for all $\epsilon>0$ there is a neighborhood $U$ of $x$ such that $f[U]$ is contained in the ball of radius $\epsilon$ around $f(x)$.  Since that ball has diameter $\leq 2\epsilon$, this means $\omega_f(x)\leq 2\epsilon$.  Since $\epsilon$ is arbitrary, this means $\omega_f(x)=0$.  Conversely, if $\omega_f(x)=0$, then for all $\epsilon>0$, there is an open neighborhood $U$ of $x$ such that $f[U]$ has diameter $<\epsilon$.  Since $f(x)\in f[U]$, this means $f[U]$ is contained in the ball of radius $\epsilon$ around $f(x)$.  Since $\epsilon$ is arbitrary, this means $f$ is continuous at $x$.

redpajama_set_name: RedPajamaStackExchange


text:
Das deutsche Wort Testament stammt vom lateinischen testamentum. Dessen Ursprungsbedeutung ist "rechtsverbindliche Verfügung", etwa als Regelung für den Erbfall.

Im christlichen Kirchenlatein ist testamentum eine Begriffsübernahme des griechischen diatheke (διαθήκη) des Griechischen Alten Testaments, dem ursprünglichen christlichen Schrifttum in griechischer Sprache. Testamentum und diatheke sind Übersetzungen des hebräischen Begriffs berit aus der Torah.

Eine diatheke war ganz allgemein ein Vertrag, der von einer Seite angeboten wurde, und von der anderen Seite akzeptiert oder abgelehnt, aber nicht geändert werden konnte.
Diese Bedeutung ist gemeint in der Bezeichnung der beiden Teile der christlichen Bibel als Altes Testament und Neues Testament und in den Abendmahlsworten der lutherischen Bibelübersetzung und Liturgie: "Das ist das neue Testament in meinem Blut" bzw. "Das ist mein Blut des neuen Testaments".

Am Berg Sinai bot Gott einen Bund (berit) an, den das ganze Volk Israel annahm und bezeugte.

Literatur
 Augustin Edmund Engelbrecht: Kurzgefaßte Erzählungen aus der biblischen Geschichte des Neuen Testaments. Druck und Verlag von Friedrich Winkler, Passau 1838.

Einzelnachweise

Biblisches Thema

redpajama_set_name: RedPajamaWikipedia


text:
Fla. State Attorney Aramis Ayala asks officers why they stopped her
| Giovedi, Luglio 13, 2017
On Wednesday, the Orlando Police Department (OPD) released the bodycam footage from an officer who pulled over State Attorney Aramis Ayala on June 19. Rick Scott and triggered a backlash. After Ayala handed an officer her ID, told them she was a state attorney, and asked a few simple questions about why she was stopped-because she was doing nothing wrong. [Full Article...]
Grassley: Hope for bipartisan health care bill based on 'dreaming'
While the House has passed health care legislation, a bill that was largely crafted without the input of rank-and-file members has stalled in the Senate. The plan scoffed at by several Senate Republicans would keep parts of Obamacare intact, but strip away much of its funding. "And they're being asked to vote for something that we never promised". [Full Article...]
Palermo, uccello con testa mozzata davanti a scuola Falcone. "Intimidazione"
Il 19 luglio, in occasione del 25esimo anniversario della strage di via D'Amelio , in città arriverà la ministra dell'Istruzione Valeria Fedeli: nei giorni scorsi ha contattato la dirigente della scuola, Daniela Lo Verde , per esprimerle sostegno e vicinanza e assicurarle che "la statua sarebbe stata subito restaurata". [Full Article...]
Tottenham must battle Liverpool for Angel Correa
Atleti can not sign players until the turn of next year due to their transfer embargo being upheld by the Court of Arbitration for Sport last month. Atletico signed the Argentine hitman from San Lorenzo in 2014, splashing out £6.5m and handing him a contract until the summer of 2019. The Argentinean forward, who netted four times in the Copa del Rey, was behind Antoine Griezmann and Fernand... [Full Article...]
Lightning hits bride's father during wedding speech
Nadeau looked down just in time to see his right hand light up. JP Nadeau from Canada told his local radio station that as he began his knockout monologue to a room full of family and friends, a lightening bolt struck, and surged through his hand as he toasted his daughter and her new husband. [Full Article...]
Student halls evacuated over bomb scare
The emergency operation was just precautionary, Haas said. Campus police said the threat was called in by an unknown person. Around 12:15 a.m. Thursday, some students were allowed to reenter their residence halls as police continued to inspect the scene. [Full Article...]
Cristiano Ronaldo posts picture of his extended family
The Portuguese was surprised by their arrival as he spent time with his girlfriend Georgina, his mother Dolores and a number of close friends and family. The process was said to have lasted around an hour and a half and a member of the striker's entourage was the one who dealt with the administrative work and showed the agents the documents they required. [Full Article...]
Buhari recuperating fast, will be back shortly - Osinbajo
He is recuperating very quickly and he is doing very well. The looming shake-up has caused anxiety among members of the Federal Executive Council (FEC). On when they will be sworn in, he said: "Well, I'm not in a position to say anything". I don't have a date in mind but I'm sure very very soon, we will do that. [Full Article...]
BOK revises up growth forecast to 2.8%
The core inflation, which excludes products in food and energy sectors, was estimated at 1.7 percent in 2017 and 1.9 percent in 2018. Lee said there were downside risks such as trade protectionism in major trading partners such as the United States and geopolitical uncertainties over North Korea's nuclear and missile threats. [Full Article...]
Albany 6-year-old dies after being hit by truck
At 9:55 p.m., troopers were called to the scene of a crash involving a commercial motor vehicle and a pedestrian on I-5 near Milepost 230, about three miles south of Albany. OSP officials have declined to release the name of the victim or other people involved. The Oregon State Police and Linn County Sheriff's Office, including a canine unit, immediately began searching for the boy. [Full Article...]
Kohli, Anushka backseat selfie gets 1 Million likes in 12 hours only
Indian cricket team recently won the five-match ODI series against West Indies with Captain Virat Kohli scoring yet another global century in the last match. In an interview last month, Kohli had opened about his relationship with Anushka. The two are chilling away in NY and of course, there to attend IIFA 2017. [Full Article...]
Family Planning: Empowering People, Developing Nations
Duruiheoma said: "As the theme of the 2017 World Population Day shows, family planning is not only about saving lives, but also empowering people and developing nations. The pledges came at the FP2020 Summit in London, where more than 60 governments and partners have committed to spend at least $2.5 billion by 2020. [Full Article...]
Bryce Harper's fantastic All-Star Game cleats showcase "Miami Bryce"
Baseball Commissioner Rob Manfred says Aaron Judge is the kind of player "who can become the face of the game". The Washington Nationals slugger said Monday he would prefer the teams be drafted by players regardless of league affiliation. [Full Article...]
Video shows arrest of National Football League player outside Sharonville water park
The video is 2:31 in length and begins with officers approaching Washington's and loudly yelling at him as he has a hand gun in his lap. In video of the arrest obtained by TMZ, the tense situation was captured on tape and shows officers surrounding Washington's vehicle, telling him to keep his hands out of the auto and exit the vehicle. [Full Article...]
WH: 'Russia a Cyber Threat,' But We Need Discussion
Trump said that Putin "vehemently denied" the conclusions of American intelligence agencies that Russian hackers and propagandists tried to sway the election in Trump's favor. But Trump would not say whether he believed Putin, tweeting only that he's "already given my opinion". "Questions were asked about why the CIA & FBI had to ask the DNC 13 times for their SERVER, and were rejected, still d... [Full Article...]
China offers help on Kashmir says situation has drawn global attention
China on Wednesday in unusual comments on Kashmir said the recentfiring across the LoC between India and Pakistan had "attracted global attention", but was silent on the terrorist attack on Amarnath pilgrims. "It is a long border, as you know no part of the border has been agreed upon on the ground. It is likely that from time to time there are differences". [Full Article...]
12 heridos y cinco vehículos destruidos deja explosión en Lima
El trabajador aseguró que labora hace años para dicha empresa y que usualmente reparte balones de gas en viviendas de esa zona del distrito. La explosión de un vehículo que transportaba gas licuado de petróleo (GLP) fue el causante de detonaciones consecutivas que alertaron a los ciudadanos y a las autoridades. [Full Article...]
Lithia Motors, Inc. (LAD) Releases Q2 Earnings Guidance
The business had revenue of $2.24 billion during the quarter, compared to analysts' expectations of $2.22 billion. Point72 Asset Mngmt LP stated it has 0.04% of its portfolio in Lithia Motors Inc (NYSE: LAD ). Lithia Motors , (LAD) transacted up 7.18% throughout intraday market trading on early Wednesday, topping $97.50. A number of research analysts have issued reports on the company. [Full Article...]
NSE technical glitch matter of serious concern: Fin Min
According to the stock exchange, the technical glitch has been resolved. Nifty surged 105.25 points, or 1.09%, ending at 9771.05. In the afternoon, Sebi, in a statement, said it was closely monitoring the situation. The rally saw total market capitalisation hitting the $2 trillion mark. Led by gains in the stock market, market capitalisation (m-cap) of BSE listed companies rose to Rs 1,30,13... [Full Article...]
Floods and landslides in northeastern India claim more lives
Assuring all possible support from Centre, the Prime Minister informed Khandu that the PMO is continuously monitoring the situation in Arunachal Pradesh and has assigned Principal Secretary to PM, PK Mishra for the goal. While a landslide was reported in Arunachal Pradesh which hit eight dwellings in Papum Pare district, flood and bridge collapse have been reported in Assam and Nagaland respec... [Full Article...]
Natural disaster off N.Korea 'no way' caused by nuke test
Sumatran tiger triplets born at OKC Zoo
Costa Rica, Canada draw to stay level on points atop Group A
McIlroy feeling good ahead of Scottish Open
Hamilton man accused of Kitchener stabbing death arrested in Texas, police say
Vladimir Putin warns against 'losing cool' on North Korea
Physical education for girls to be introduced in Saudi Arabia
Voters trust the media slightly more than the scandal-ridden Trump administration
U.S. to renegotiate Korea trade deal
Afghan teens finally approved to attend United States robotics competition


Steve Moskos' Blog - C. STEVEN MOSKOS, P.A.
Arbitration is a way to resolve disputes. Its proponents say it is faster and cheaper than using the court system. Its detractors say it protects businesses from big verdicts that would change the wrongful way a business operates.
"All you really need to know about today's process is that it's the product of years of conceptual monkey-wrenching by corporate lobbyists, Congress, the Supreme Court and hired-gun lobbying firms looking to milk the system for steady profits. First and foremost, these fixers have turned a voluntary process into the exact opposite: mandatory. Let's look at this mess.
Unlike courts, arbitration is not a public system, but a private business.
Far from being neutral, "the third-party" arbitration firms are - get this! - usually hand-picked by the corporation involved in the case, chosen specifically because they have proven records of favoring the corporation.
The corporation also gets to choose the city or town where the case is heard, allowing it to make the case inconvenient, expensive and unfair to individuals bringing a complaint.
Arbitrators are not required to know the law relevant to the cases they judge or to follow legal precedents.
Normal procedural rules for gathering and sharing evidence and safeguarding fairness to both parties do not apply in arbitration cases.
Arbitration proceedings are closed to the media and the public.
Arbitrators need not reveal the reasons for their decisions, so they are not legally accountable for errors, and the decisions set no legal precedents for guiding future corporate conduct.
Even if an arbitrator's decision is legally incorrect, it still is enforceable, carrying the full weight of the law.
Does this mean that arbitration doesn't have any place in your judicial system? No. What it means is that one must be suspicious of a process that is forced on someone before he or she even knows there is a problem. The next time you buy a car, apply for a loan, or apply for a credit card, tell the business you want the mandatory binding arbitration provision removed from the contract. See what happens. If it is such a great process, why won't businesses wait until an unresolved problem appears? Then the business can bring up arbitration if it wants to. Then both sides can decide whether arbitration is best for them.
Can I return a car within 72 hours?
If a car I buy is defective, can I return it within 72 hours?
Does Carfax tell me everything I need to know before I buy a car?
We appreciate you coming to our website, however, the information we have here should not be construed as formal legal advice nor should you believe we have established a lawyer/client relationship. This website is for informational purposes only. Contacting C. Steven Moskos, PA through this site does not create an attorney-client relationship between you and C. Steven Moskos, PA. This law firm must investigate your case, evaluate it, do a check for conflicts-of-interest, and sign a written retainer agreement with you before you become a client.
In addition, the information contained on this website is not intended to guarantee that we can produce the same results for you that we have produced for others. Each case is different and must be evaluated on its own merits.


We believe that development is about increasing possibilities and capacities of individuals to make the most of their potential to live as full resourceful persons and together with others create caring, supportive and accountable societies.
Community and national development to us is about respecting, protecting, promoting and meeting basic rights of people including that of children for human dignity, development and survival. We believe in a well-integrated society that recognises children in every sector and creates the necessary space and opportunity to respond to their developmental needs. We also believe that every child, given the opportunity and protection by society, can optimise his/her God-given abilities and grow to lead an independent, responsible adult life.
CRRECENT believes that rights and responsibilities of children living under challenging circumstances can be realised through research, evidence-based policy advocacy, sustained and purposeful conscientisation of children and individuals living with deprived children and their institutions supported by creating concrete alternative that challenge and address structural causes and consequences of child poverty.
CRRECENT re-enforces the principle of non-compartmentalization of the child's development and brings to the fore the multi-sectoral nature of children's issues and the urgent need to tackle such at both Policy and programmematic levels. CRRECENT targets children and youth, adults working with children and policy makers in its operations. Beneficiaries under our programmes include: (a) Vulnerable and challenged children (0-18) in the areas of child rights; child survival and development; education and protection. (b) Challenged youth (19-25) in life changing programmes (education and skill development) to prepare them for successful independent live. Other beneficiaries comprise of: Government Ministries such as Education; Health; Employment; Social Welfare (Correctional Centres and Remand Homes for Boys & Girls); and Local Government and Administrations.
CRRECENT's focus ensures that the rights of children living in challenging circumstances are respected, promoted and fulfilled and that state institutions become responsible in ensuring the rights of children.


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Every good business is built on top of a compelling Value Proposition, the real reason why customers find you desirable.
If you can learn how your customer thinks, you can create offers that entice and delight, leading to more sales and a loyal audience.
This is a fascinating field, and in these articles you can learn how to create your own great Value Propositions.


Researchers taking a second look at ancient sediments have concluded that they do not represent the earliest traces of the rise of sun-fuelled photosynthesis/oxygen-producing organisms on Earth as previously thought, reports ScienceNOW website.
It says the findings come as a relief to some scientists, whose data had been in conflict with an earlier study with the same sediments that suggested oxygen first emerged in the planet's atmosphere some 300 million years later.
According to the geological record, oxygen first rose in Earth's atmosphere about 2.4bn years ago. The problem with using fossils to track down the organisms that gave rise to the oxygen is that fossils can take you back only about 2bn years out of the planet's 4.5bn.
To figure out what was stirring before that, scientists must rely on traces of chemicals produced by biological processes and, by inference, in certain environmental conditions.
That's what happened in 1999, when researchers examined samples of 2.7bn-year-old shale from Western Australia.
They found certain types of hydrocarbons that indicated oxygen-producing bacteria had existed at the time. But the discovery also created a conundrum, because it placed the rise of photosynthesis some 300m years before chemical studies of ancient rocks indicated oxygen began to spread in Earth's atmosphere.
This discrepancy continues to bedevil researchers, who have struggled to mesh their data with evolutionary models.
An Australian team, writing in Nature, analysed microscopic solidified oil droplets contained in the shale, the researchers inferred the composition of the organic materials present as the shale formed.
The analysis clearly shows the hydrocarbons identified in the 1999 study could not have been derived from bacteria contained in the sediments, says geochemist and lead author Birger Rasmussen of the Curtin University of Technology, Perth, Australia.
Instead, they probably represent contaminants introduced from younger sedimentary rocks that somehow got mixed in with the shale layer or by the drills the researchers used to extract the samples, he says.
The conclusions are "pretty strong and a lot of eyes will be focused on how this shakes out," says geobiologist Woodward Fischer of the California Institute of Technology, Pasadena, US.
He says the previous findings had been regarded as a standard time measurement for both the rise of oxygen and for photosynthetic organisms, and, therefore, any conflicting data produced by other researchers had been regarded as suspect.
But overall he's convinced that the new results ultimately "will free up people's minds again" to find the other sources of early oxygen.
Over thirty years ago, I heard that certain amino acids were found in an uncombined state from what are known as 'white' smokers, low temperature under sea hot springs. My immediate reaction, "That was where life originated" i.e. in a high energy environment with unstable compounds coupled with aluminoslicates and silica where early life would mimic, crystal growth. (It was only about a decade later that John Corliss from Corvallis actually wrote up the idea in a professional paper instead of just walking away from it.) With such a rich source of energy, life would have originated as a way to economically transform complex compounds to lower energy states.
According to my beliefs, primitive life from the undersea hotsprings drifted upsection where they encountered the photic zone. Their primitive DNA was promptly fried by ultraviolet radiation. The predictable result—mutation and natural selection. The natural selection would favor compounds that could shield the cell's DNA from disruption. The most likely candidate: Some kind of primitive chlorophyll that would intercept the high energy ultraviolet radiation and transfrom it into something that would not damage early life's proto DNA.
In this model of the development of chlorophyll, life around the high energy environment of chemosynthesis around the hot springs had no need for food because all the food that was needed could be provided by chemosynthesis. It was only when life drifted up into the chemosynthetically challenged surface waters that creating food was essential for propogating life beyond the range of chemosynthesis related life. First early life put on the sun block, and only later decided that by products of the sun block were mighty tasty.


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Knock Knowes is a spacious detached family house with adaptable accommodation. The house has a "granny annexe" which has, in the past, been used as B&B accommodation. To the rear of the house is a workshop/garage.


It looks as though the lie has gone around the world and the truth needs to get its skates on (again). This blog is no friend of the numerous attempts to smear the British National Health Service so let's share this around and make sure people can read arguments both for AND against the current system – so they can make up their own minds.
No institution is perfect, but the Tories want to destroy our NHS and we won't get it back from New Labour if the Tories and their little yellow friends the Lib Dems break it.


text:
Brawlidays: new skins, snow theme and boss fight
Howdy Brawlers! The long awaited Brawlidays are finally here! Today Supercell has published all the information about this special Christmas event. Here is a summary of everything we know to date.
The most publicized and expected feature of the Brawlidays will surely be three new Christmas skins. These masks will only be available for limited amount of time After January 3, none of them will be available, so get them while you can!
First, we have Santamike, a skin for Dynamike. It definitely fits our favorite white beard explosive miner! His attacks have also been reskinned: he throws candy canes with his main attack and an explosive Christmas tree with his sper. This mask is available for 80 gems
Second, we have the Lil's assistant penny skin. With this skin, Penny looks like a festive and mischievous elf. The skin also extends to your dog, which looks like a Christmas gift. This skin can be bought by 80 gems also.
Finally, the most anticipated skin of the group has to be Red Nose Nita. There is a reason for that for sure: this skin offers a complex reworking of the look of Nita and her bear, and they both have a Rudolph reindeer costume. This skin is also the most expensive of the three, since it costs 150 gems.
Christmas says sales, and this is also true for Brawlidays. 4 special offers They currently run in all Brawler stores in the world.
To start, the three new masks are available at reduced prices at this time. Santamike and Lil 'Helper Penny cost 59 gems, while Red Nose Nita can be purchased for 109 gems
The fourth special offer is a package that costs real money. by $ 4.99 USA UU. OR $ 6.99 CAN, you can buy the holiday package, which contains 2 Mega Boxes and 170 gems, for a total value of 4x.
The offers of masks will be available for 14 days, but the vacation package will only be available for 6 days, which allows us to think that there could be more in the future.
The last thing Brawlidays present is the new map, Snow fort. It is a clone of the Stone Fort map and can be played in Gem Grab. It presents a new snowy theme, with a floor made of ice, pale blue grass and multiple Christmas references, as gifts instead of boxes. Hopefully, we will see more maps with this theme for the rest of the Brawlidays!
Boss Fight
We could believe everything we saw that Brawlidays has to offer. With new masks, many store offers and a new map theme, Supercell is already giving us many gifts. But there is one more thing we haven't seen yet.
Boss Fight It's a new ticket game mode: remember that the old Boss Fight has been renamed Big Game! In this PvE game mode, compete with friends or randoms against giant enemies, similar to Boss Rush mode. That means that once a boss is defeated, you won't have to fight him again! Bosses become increasingly challenging once you reach higher levels. Once you have defeated a boss, you can fight them again for free if you play with friends who are at a lower level. Do not win any trophies with this mode, but you may be able to grow a load of coins or sled boxes.
For this Brawliday season, Supercell is really doing everything possible with their gifts. With all this new content, we can only hope that it will be something that will be repeated in the coming years. That is all for now; If you have any questions, feel free to leave it in the comments below.
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The Bible tells us that Samuel "grieved" (abal) over Saul. But Yahweh told Samuel that the time for grieving was over, and that it was time to appoint a new king.
The Amish resist certain aspects of "moving on." I appreciate the Anabaptist resistance to the inhumane features of "progress," the Anabaptist call to simplicity and fidelity to ancient traditions. But why stop with the 19th century? Why not go to a period prior to buggies, ovens, cupboards and battery-operated adding machines? The operative word here, as Donald Kraybill so ably demonstrates, is the German word Gelassenheit, or "yieldedness"—to God's loving, providing and guiding will. But sometimes what is perceived as Gelassenheit is actually a stubborn resistance to the inevitability of change.
The gospel proclaims an alarming fact about historical movement—it is what God is all about. The entire Bible hinges on one undeniable reality: reality is God's workshop. God doesn't give Abraham a set of beliefs but an event (a smoking fire pot) and a rite (circumcision). And God gives the Christian church a son—a child born of a woman whose reputation was stained, and reared by a father who surrendered his status as a tsadiq or "righteous man." Yet this son does not just teach the gospel: he embodies it.
In acting this way God sanctifies history, making it something to embrace instead of resist. When Samuel resists he hears the voice of God directing him to a future that will be better. That future will include David the shepherd boy. Like all shepherds, he is often on the move. As the author of Psalm 23, David the shepherd lies down, is led beside still waters, walks through the shadow of death and sits in the presence of his enemies. David will do whatever it takes to guide his sheep even as he remembers that Yahweh is his shepherd, guiding him.
Then another shepherd will arrive. Jesus the Good Shepherd will be the Light of the World, removing darkness and trumping, for example, the darkness of the man born blind. Like Samuel, the disciples will "get stuck" because they'll wonder whose sins have made the man blind. But Jesus, pushing them into the "Shepherd's era," will lead them to see that simplistic correlations from the past (sin leads to curse, obedience leads to blessing) do not always work.
He will guide them with his light, and when that light is turned on, three things will happen. First, those who live in that light "try to learn what is pleasing to the Lord." Samuel was stuck for some time in wanting Saul's era to be the kingdom era, but God gave him a horn of oil to search for the Shepherd's era. It does no good to apply more and more oil to the old era, God said. It is gone; it is history. We please God by moving on.
We do this too by taking no part in the "unfruitful works of darkness," but instead exposing them. Like many others, I am deeply saddened by Americans' tendency to gloat triumphally in its victories. I am also saddened by Christians who, instead of weeping over current world affairs, have picked up a new sword of Constantine, a wicked instrument of triumphalism.
We need what John Howard Yoder calls the "politics of Jesus" and what Stanley Hauerwas calls the "peaceable kingdom." Archbishop of Canterbury Rowan Williams says it well: "From now on, all that can be said of God's action in the past or the present must pass under the judgement of this fact [the cross]."He also says, "God is known in and by the exercise of crucifying compassion; if we are like him in that, we know him." These theologians are calling us out of the old era of warfare, the Saul era, into the Shepherd's era of justice, peace and love.
This future kingdom is marked by "justice," a word that is fast losing its robust Christian profile. It has, as Flannery O'Connor said of another word, "a private meaning and public odor." Some use the term in the sense of "retribution"(bring them to justice), and some in the sense of "rectification"(give the victims and the marginalized an equal opportunity). Neither of these senses is adequately Christian. The Christian sense of "justice" is "what is right before God and others." And, according to Jesus' own creed, what is "right" is to love God and to love others (Mark 12:29-31). In the Christian sense, justice means providing our world an opportunity to love God and to love others.
I am intrigued by the emphasis on call in 1 Samuel 16. Because I am a theological educator, I am even more fascinated by the role each of us can play in nurturing someone's sense of call. Saul and David are the key "called" protagonists in the story. But it is Samuel who carries, clarifies, and extends God's call.
David stares at us out of a mirror and shows us our capacity for sinning.