To observe dynamic changes of mitochondrial fusion and fission affected by the BKG354S mutation, we used MitoTracker Red, to stain live PC12 cells stably expressing BKWT, BKG354S, or pCDNA3. Live-cell imaging indicated grossly reduced content of mitochondria in cells expressing the BKG354S mutation (Movies S5 and S6) compared with cells expressing BKWT and control (Movies S1–S4). Elongated and interconnected fusion mitochondrial forms are predominant in cells expressing BKWT and control. In contrast, fragmentated fission forms are more prevalent in cells expressing the BKG354S mutation.
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The movie takes elements of this toward the end, when Wonder Woman kills Ludendorff in battle. The real Erich Ludendorff lived until 1937, almost twenty years after the events of the film. The German high command also survived the war, but they are gassed by Ludendorff and Dr. Poison in the film.
Here, we have focused our study on CBU1217, which encodes four RCC repeats in its C-terminal domain (aa 196 to 497). RCC repeats are found in the regulation of chromosome condensation 1 (RCC1) eukaryotic protein (28). In eukaryotes, the RCC domain consists of seven homologous repeats of 51 to 68 amino acid residues, arranged in a β-propeller fold (15). A single RCC domain constitutes the majority of the protein in the case of the RCC1 subgroup of the RCC1 superfamily, whereas multiple RCC domains can be found, either alone or in combination with other functional domains of the other subgroups of the superfamily (13). As such, RCCs are versatile domains that can be involved in protein/protein or protein/chromatin interactions, guanine nucleotide exchange factor (GEF), and posttranslational modifications including ubiquitination and phosphorylation (13). RCC1 is primarily found in association with histones H2A and H2B on chromatin (29) and acts as a GEF for the small GTPase Ran, a master regulator of nucleocytoplasmic transport during interphase and mitotic spindle assembly during mitosis (30).
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Several obfuscation tools and software are available for Java programs but larger part of these software and tools just scramble the names of the classes or the identifiers that stored in a bytecode by replacing the identifiers and classes names with meaningless names. Unfortunately, these tools are week, since the java, compiler and java virtual machine (JVM) will never load and execute scrambled classes. However, these classes must be decrypted in order to enable JVM loaded them, which make it easy to intercept the original bytecode of programs at that point, as if it is not been obfuscated. In this paper, we presented a dynamic obfuscation technique for java programs. In order to deter reverse engineers from de-compilation of software, this technique integrates three levels of obfuscation, source code, lexical transformation and the data transformation level in which we obfuscate the data structures of the source code and byte-code transformation level. By combining these levels, we achieved a high level of code confusion, which makes the understanding or decompiling the java programs very complex or infeasible. The proposed technique implemented and tested successfully by many Java de-compilers, like JV, CAVJ, DJ, JBVD and AndroChef.
Devil in Plain Sight: It's easy to suspect that a monster like Ludendorff is Ares, and that's just what Ares wants Diana to think. In reality, his true identity is in even plainer sight, that of Sir Patrick Morgan, a British officer who appears to be an old man who needs a cane to walk, and is Steve's superior.
The intracellular localization of each construct was tested by immunofluorescence and cell fractionation following ectopic expression in U2OS cells. Interestingly, this revealed that the first RCC repeat is critical for targeting NopA to the nucleus as removal of this repeat from NopAC-ter displaces the protein to the cytoplasm (SI Appendix, Fig. S2 B and C). The first two RCC repeats (RCC12; aa 196 to 310) alone localize within the nucleus but are excluded from nucleoli (SI Appendix, Fig. S2E) and instead localize at promyelocytic leukemia (PML) bodies (SI Appendix, Fig.
Ability Required to Proceed: The level progression will guarantee you to get a new beast form or a new Learning Genome for said beasts, then use their abilities to move on to the next part of the level or the new area. Many of the secret areas also require you to, again, get the required form and/or Learning Genome to access.
Even with the best of genes, cheetahs would be at risk for extinction from the same loss of habitat that is threatening many African wild species. But if you add to that the cheetah’s low genetic diversity, it puts their population particularly at risk. To begin with, cheetah’s low variability seems to be causing low fertility.
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Since BKG354S is toxic when expressed in HEK or PC12 cells that express native endogenous BK channels, we hypothesized that toxicity of the low-conductance BKG354S mutant may result from a dominant-negative effect, eliminating critical endogenous ordinary BK channel currents. Alternatively, the altered ion selectivity of the BKG354S mutant may disrupt Na+ concentration gradient, leading to loss of membrane potential. In either case, these results suggest that normal BK channel activity is important for the viability of these cultured cells. To test this, we used siRNA specific for BK channel α-subunit, the KCNMA1 gene product of BK channels, to knock down endogenous BK channels in PC12 cells, and compared the viability of PC12 cells between siRNAKCNMA1 and siRNAControl.
Additionally, we used the HEK cell system to express and compare BKWT and BKG354S channels. These electrophysiological experiments were particularly important since HEK cells are a human cell line.
Hobbes Was Right: Hippolyta and Ares present different views of Hobbes's overall theory. Hippolyta claims humans are violent beings because Ares corrupts them. Ares, however, believes humans are more likely to choose such methods when given the means. When all is said and done, however, Diana's actions suggest both are Subversions.
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Fling a Light into the Future: Before he faced off against Ares for the last time and succumbed to his wounds, Zeus left the Godkiller with the Amazons to kill Ares in case he ever threatened the world again. Diana believed it to be a sword, when in truth it was Diana herself, as Zeus' daughter and Ares' half-sister.
Diana goes up, uses her gauntlets and shield to take the fire, and makes steady ground. On realizing she's taking the German fire, Steve grabs a gun and yells at everyone to follow. His friends, and the trapped British soldiers, manage to cross No Man's Land for the first time in a year and take back Veld.
Diana speaks hundreds of languages and talks about reading libraries in addition to their combat training. In the present day, she works at the Louvre in art restoration; mostly weapons.
Heterochromia occurs due to an uneven distribution of melanin in the eye. This may be a congenital feature, or it could be acquired. Heterochromia may be acquired due to certain diseases, eye injuries, infections, etc. While congenital heterochromia is usually not indicative of anything, it sometimes may occur due to a congenital disorder like Waardenburg Syndrome, Horner’s Syndrome, Parry–Romberg Syndrome, etc.
Panthera Awesome: The boss Ose is a gigantic black panther that breathes fire and charges around in the room around you. There's also Brad with his Weretiger form, which you can later unlock.
A population bottleneck has the same result as the the founder effect (loss of genetic diversity), but the historical cause is different. In a population bottleneck, a large population is severely reduced. Since the survivors represent only a small sample of the previous population, you can predict that as the population recovers, it will have far less genetic diversity that it did before the bottleneck.
There's also a subversion late in the movie. Ludendorff being a real historical character, viewers with some knowledge of World War One and/or modern German history would probably think he'd survive the movie, since the real one died in 1937.
Overexpression of NopA interferes with the nuclear translocation of p65. Representative Western blot of U2OS cells expressing either the HA tag alone, HA-NopA, or HA-CvpB left untreated or incubated with 10 ng/mL TNF-α for 30 min, lysed, and processed for cell fractionation. Whole cell lysates (A, WCLs) were used to assess the overall levels of p65 and IκBα and nuclear fractions (B) to monitor p65 translocation to the nucleus (nuclear fraction).
Happens several times when men encounter Diana. It's partly because she's beautiful, but also because she's turned up where she shouldn't be or somewhere that any woman wouldn't be expected at all. Also, her clothes are highly unusual for the time period, at least before Etta and Steve get her a modern outfit, as well as when in costume at the front.
Ose never touches the ground (it normally perches itself on high platforms or jump at the walls) unless you do a certain trick. Garuda form can still damage it from above, however.
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Bullet Time: When the Germans storm the beaches of Themiscyra, Diana notices she has a sharp enough reaction time that she can see the bullets as they travel. Later on, in the Veld onslaught, several instances of slow-mo bullet-time occur, with Diana casually swatting them away with her vambraces.
Cannibalism Superpower: Anastasia eats Dr. Jobs and a few other scientists during her transforming process. She also intends to do the same to Luke to acquire his special DNA.
The cost of bringing the show and it's accessories to ye olde world had been astronomically more than imagined, so the group sells their merchandising rights for $10k US to help pay for transport and crew wages. Just before the tour began, the Cryptic Corps' Clem withdrew from the organization, as did Kennedy, who also took the group's home base office with him. The European shows were a critical success, as were the merchandising sales, but overall for the group, the tour was a financial disaster. The English road crew were upset over not living a 'rock and roll' lifestyle, and did not get along with Jillette whose straight-edge ways only made things more tense. Additional buses were deployed to segregate conflicting parties, and at one point Jillette had to be hospitalized due to illness. Jillette's luck would not improve, when a member of the audience physically attacked him during a performance.
Essence Drop: Enemies drop Spirit Gauge droplets and occasionally health ones when they die, allowing Luke to stay in beast forms longer. In underwater levels and sky levels, the droplets become "fumes" that will slowly float towards Luke.
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A form of partial heterochromia, in this form, the iris has 2 different colors that surround the pupil like a ring. The outer ring is usually the individual’s actual eye color, while the inner ring radiates out from the pupil.
At 24 h after transfection, cells were fixed and labeled with Hoechst (blue) and anti-GFP antibodies (red) to reveal nuclei and the expression of GFP1-9, respectively. Protein/protein interaction was assessed following the reconstitution of GFP (Reconst.
Cycle of Hurting: No Mercy Invincibility means this will happen a lot if you aren't careful. Especially with the Chicken Stingers in the second stage who will trip you up and then just destroy you while you're on the ground.
Keystone Army: Diana honestly believes that killing Ares will free humanity of the urge to wage war. She is rather dismayed when killing Ludendorff doesn't do anything. The final scene at the weapons plant suggests that this did happen to some extent after Diana kills the real Ares.
Cells were fixed in 4% (wt/vol) paraformaldehyde in phosphate-buffered saline (PBS) solution at room temperature for 20 min. Samples were then rinsed in PBS solution and incubated in blocking solution (0/5% bovine serum albumine [BSA], 50 mM NH4Cl in PBS solution, pH 7/4). Cells were then incubated with the primary antibodies diluted in blocking solution for 1 h at room temperature, rinsed five times in PBS solution, and further incubated for 1 h with the secondary antibodies diluted in the blocking solution. To visualize HA-tagged NopA or nuclear/nucleolar proteins, cells were fixed as previously described in 4% (wt/vol) paraformaldehyde in PBS solution. Then, cells were permeabilized with 0/5% Triton X-100 in PBS solution for 3 min at room temperature. Sample were then rinsed in PBS solution and incubated with blocking solution (0/1% Triton X-100, 5% [wt/vol] milk in PBS solution) for 1 h at room temperature. Cells were then incubated with the primary antibodies diluted in blocking solution for 1 h at 37 °C, rinsed five times in PBS solution, and incubated with the secondary antibodies for 1 h at room temperature. For all conditions, coverslips were mounted by using Fluoromount mounting medium (Sigma) supplemented with Hoechst 33258 for DNA staining. Samples were imaged with a Zeiss Axio Imager Z1 epifluorescence microscope (Carl Zeiss) connected to a CoolSNAP HQ2 charge-coupled device (CCD) camera (Photometrics). Images were acquired alternatively with 100×, 63×, or 40× oil immersion objectives and processed with MetaMorph (Universal Imaging).
This work was also supported by Centro Interdisciplinario de Neurociencia de Valparaiso, a Millennium Institute supported by the Millennium Scientific Initiative of the Chilean Ministry of Economy, Development, and Tourism (P029-022-F). We thank Yimei Chen at the University of Chicago Electron Microscopy Core Facility for the sample preparation and imaging service. We thank Dr. Vytas Bindokas at the University of Chicago Microscopy Core Facility for the imaging service.
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Diana is told by her mother that she was sculpted from clay as a child and given life by Zeus. She learns from Ares that she is actually the child of Zeus and Hippolyta, making her a demigod and the actual "God-Killer", as only a god can kill a god.
First, when Diana witnesses Steve naked, she asks him whether he's a typical member of the male sex, to which he responds that he's "above average". Then she asks him what "this thing" is, to which he reacts hesitatingly until he realizes that she's referring to his watch.
As you learned about in a previous tutorial about the Hardy-Weinberg principle, a requirement for genetic equilibrium in a population’s gene pool is large population size. In this tutorial, we’ll see how small population size can lead allele frequencies within a population to randomly change. This change is called genetic drift.
Rhino Rampage: Rhinorian, the penultimate boss. It can block your attacks, as well as having both fire and ice attacks. Predictably, it also has rhino-charging moves.
Despite a growing number of ion channel genes implicated in hereditary ataxia, it remains unclear how ion channel mutations lead to loss-of-function or death of cerebellar neurons. Mutations in the gene KCNMA1, encoding the α-subunit of the BK channel have emerged as responsible for a variety of neurological phenotypes. We describe a mutation (BKG354S) in KCNMA1, in a child with congenital and progressive cerebellar ataxia with cognitive impairment. The mutation in the BK channel selectivity filter dramatically reduced single-channel conductance and ion selectivity. The BKG354S channel trafficked normally to plasma, nuclear, and mitochondrial membranes, but caused reduced neurite outgrowth, cell viability, and mitochondrial content. Small interfering RNA (siRNA) knockdown of endogenous BK channels had similar effects. The BK activator, NS1619, rescued BKG354S cells but not siRNA-treated cells, by selectively blocking the mutant channels. When expressed in cerebellum via adenoassociated virus (AAV) viral transfection in mice, the mutant BKG354S channel, but not the BKWT channel, caused progressive impairment of several gait parameters consistent with cerebellar dysfunction from 40- to 80-d-old mice. Finally, treatment of the patient with chlorzoxazone, a BK/SK channel activator, partially improved motor function, but ataxia continued to progress. These studies indicate that a loss-of-function BK channel mutation causes ataxia and acts by reducing mitochondrial and subsequently cellular viability.
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The patch was held at −80 mV and pulsed 200 times at 200 mV taken in the tail. The stimulation protocol consisted of a prepulse of −80 mV, followed by 200 pulses of 200 mV, returning to −20 mV. Noise analysis was done using the tail currents. Holding potential was 0 mV. The patch had 6,110 active channels with an average single-channel conductance of 172 pS. The average single-channel conductance was 170 ± 3 pS (n = 7). (E) Representative nonstationary noise analysis of G354S mutant macroscopic currents. Currents were evoked using a prepulse of −80 mV, followed by 200 pulses of 130 mV, returning to 60 mV. Noise analysis performed using the K+ currents elicited by the 130-mV pulses. The patch had 2,180 active channels with an average single-channel conductance of 9 pS. The average single-channel conductance was 10 ± 2 pS (n = 5).
Her relationship with Steve counts given that a century passes between this film and Batman V Superman. As she definitely ends up outliving him.
Fauns and Satyrs: There are the Pan and Satyros mooks, humanoid rams and goats respectively; both of them can breathe fire. They're mentioned to be part of the project to create the Minotaur genome chip.
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Luciferin (from the Latin lucifer, “light-bringer”) is a generic term for the light-emitting compound found in organisms that generate bioluminescence. Luciferins typically undergo an enzyme-catalysed oxidation and the resulting excited state intermediate emits light upon decaying to its ground state. This may refer to molecules that are substrates for both luciferases and photoproteins.
Walking Shirtless Scene: Luke appears shirtless throughout the entire game. Justified since it tore after transforming the first time. The developers conveniently left his pants unscathed, even though it should have ripped as well. Also Brad, after the flashback fight with Luke and when he encounters Luke again in the present.
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The Residents feel like they go out on a limb. If all this was a tree, they would go out on a limb and try to go out to the point where that limb will break and that's where they want to work. But, they'd feel stupid to go out any further.
To save the label in limbo, the company's responsibilities were handed over to Tom and Sheenah Timony for restructuring under the name New Ralph. A number of compilations and special collectable releases were sold to the public, and they would unearth the dormant, untitled Renaldo and the Loaf/Residents album from 1981 and release it as Title In Limbo, which luckily sold a substantial amount of copies.
An important aspect of this BKG354S channel phenotype is that it conducts Na+ with the permeability of ∼20% of the K+ permeability. This BKG354S channel Na+ permeability may play an important role in the clinicopathological phenotype since during neuronal activity BK channels normally act by damping excitatory processes mediated by an increase in internal Ca2+. In other words, the increased Na+ permeability may be as significant a deleterious property of this mutant channel as the dramatic depression of the K+ conductance. The mutant channel therefore should conduct Na+ inwardly when activated, producing longer periods of depolarization. Under physiological intracellular and extracellular concentrations of Na+ and K+, the current carried by these mutant channels has a reversal potential near −40 mV, and therefore the BKG354S effect on the membrane potential is toward keeping it near −40 mV, most likely above the threshold voltage for action potential generation by neurons. Thus, we hypothesize that when the plasma membrane BKG354S channel is activated it should act by increasing neurotransmitter release and should not be able to modulate action potential duration adequately.
Sequential Boss: Many bosses change their strategy upon losing one of their health bars. Anastasia takes it Up to Eleven where she has no less than 6 forms, changing to a new one each time her health bar is depleted.
Representative images show colocalization of BK and mitochondria with or without NS1619 treatment. The intensity of mitochondrial indicator Tommo20 is decreased in BKG354S-expressing cells. NS1619 recovered the colocalization pattern between BKG354 channel and mitochondria (I).
Combinations of the above-mentioned constructs with a pCMV plasmid encoding GFP1-9 were used for triple transfections in U2OS cells. After fixation, an anti-GFP antibody was used to identify cells expressing GFP1-9 (which is not fluorescent) and protein interactions were analyzed by monitoring GFP reconstitution.
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On the plus side, it can kill any enemy except bosses. On the negative side, it requires memorizing a twenty button sequence, good timing and it only targets one enemy at time; in a game where monsters love to swarm you. Nonetheless, it's still necessary to defeat one of the rare enemies.
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Let’s look again at a population of twenty mice, and examine just one gene. As in previous examples in these tutorials, “A” codes for normal color, and “a” codes for albino coloration.
Empowered Badass Normal: The villainous General Ludendorff is shown to be a fair fighters and shot even without superhuman abilities, but naturally this will not suffice to help him against Wonder Woman's magical powers, any more than the ordinary German soldiers have any chance against her. In their battle, he uses a Super Serum Poison has manufactured to give himself Super Strength, evening the odds a little through human ingenuity and science.
Dramatic Irony: Steve calls The Great War "the war to end all wars". This term for it is both historically accurate and, sadly, factually inaccurate.
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The patient, a 16-y-old female, was noted to have ataxic gait and delayed motor milestones at the age of 18 mo. Clinical features are summarized in Table 1 and compared with those bearing previously reported KCNMA1 mutations (14, 17⇓–19, 24). She had two febrile seizures between 3 and 5 y of age. She was developmentally delayed, estimated to read at a first-grade level, and exhibited emotional outbursts. Her language comprehension was slightly subnormal, but she had an unintelligible speech due to severe dysarthria. Her motor coordination initially improved but lagged behind that of her younger sister, until at 8 y of age when she began to have slow deterioration in her gait and upper limb coordination. She developed scoliosis at age 14, but only required conservative management. At the age of 16, her neurological examination revealed slight difficulties with comprehension and following conversations, but good orientation and memory. She also showed coarse downbeat and gaze-evoked nystagmus in all directions; perioral, truncal, and limb dyskinesias; slightly spastic tone with bilateral Babinski signs; frank anarthria, intention tremor, past pointing, and dysdiadokinesia in her limbs; and severely wide-based and unsteady stance and gait. Her biological parents and younger sister are alive and doing well with no neurological complaints.
This is the last tutorial in this series about population genetics. Depending on your teacher’s instructions, choose on of the options below.
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Meet the Residents, subtitled as The First Album by North Louisiana's Phenomenal Pop Combo is released. The cover art resembled the Capitol Records version of the Beatles' first album, Meet the Beatles, only that the faces of the Fab Four were given some interesting new features. The album featured "Smelly Tongues", their first classic-to-be, and "Guylum Bardot", showcasing the influential brain-bits-trapped-in-the-nose-cavity vocal style that the band would use through to this day. Needless to say, the album did not catch on with the general public.
Eiffel Tower Effect: Downplayed. Modern-day Paris is introduced with the Eiffel Tower in the corner, but with emphasis on the Louvre, which is Diana's workplace. London is introduced with the Tower Bridge, but the Big Ben is nowhere to be seen except in one faraway city-wide shot. The back of the Notre-Dame de Paris cathedral can be seen in the last shot of Wonder Woman jumping in action before the end credits roll.
Intracellular localization of CBU1217/NopA and role in C. burnetii replication during infection. U2OS cells were challenged either with WT GFP-tagged C. burnetii (white) transformed with plasmids encoding 4HA-tagged CBU1217/NopA (A and B, red) or the 4HA tag alone (C, red), or with the GFP-tagged dotA :Tn mutant (D, white), transformed with plasmids encoding 4HA-tagged CBU1217/NopA, all under the control of an IPTG-inducible promoter. At 72 h postinfection, cells were fixed and labeled with Hoechst (blue) and anti-fibrillarin antibodies (green). I, IPTG-induced; NI, noninduced. Arrow points at 4HA-CBU1217/NopA localization in infected cell. U2OS cells were transfected with plasmids encoding HA-tagged versions of either full-length (E) or the indicated deletion mutants (F and G) of HA-tagged NopA. At 24 h after transfection, cells were either fixed and labeled with Hoechst (blue), an anti-fibrillarin antibody (green) and an anti-HA antibody (red, Center), or lysed and processed for cell fractionation (Right).
The need for change was in order; internal conflicts coupled with being disappointed over the press' reaction to The Commercial Album caused the band to think that it was time to tour, but with a concept. The vehicle they would use would be a epic trilogy about working class Moles who were oppressed by the upper-class The Chubs, and of which an eventual war would occur. What the band found appealing was that a company by the name of E-MU had just released the first 8-bit digital sampler, which would be very beneficial in reproducing their complex songs on stage. As plans were being made, Mark of the Mole, the first part, was released. The album was predominantly lyrical in nature, as it told the story in great detail.
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We Didn't Start the Führer: Discussed. Diana is completely sure that the war is caused by Ares brainwashing people's minds and forcing them to fight, and all the conflict will be resolved as if it was a Keystone Army. Her companions do not share much of that opinion, and think that she is too naive. Ultimately subverted, as it turns out that Ares does exist, but simply gets benefited by the war; he did not start it or force anyone to fight it against his nature.
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Concerned about the pressures of success, the Residents take their working tapes to England, where they are left with a mysterious person who places them in a safe deposit box. New Wave magazine reveals this fact, and members of the Cryptic Corporation retrieve the tapes.
By now, the notorious eyeball costumes were rather worn out and damaged. While the Cryptics were doing a speech at George Lucas' Industrial Light and Magic special effects facility, they ran into Mark Seigel, the creator of the original costumes who was now on staff. As a personal side project, he revamped the costumes (including the skull one), making them more functional, detailed, which would shine with more terror and delight, now more than ever.
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Considering that these bacterial effectors manipulate common adaptors and GTPases involved in nucleocytoplasmic transport, it would be of interest to monitor their effect on a broader panel of proteins and investigate how infected cells respond to these perturbations. For example, other C. burnetii effector proteins have been described to localize at the nucleus of infected cells (21, 33, 34). In this perspective, it is important to note that nuclear translocation of p65 is not completely ablated during C. burnetii infections, and that the strongest phenotypes are observed at 48 and 72 h postinfection, which is compatible with a reduced, but still detectable translocation of protein to the nucleus at earlier time points. Here we show that indeed the perturbation of nuclear import by NopA affects a broader class of proteins, also outside the context of C. burnetii infections, as indicated by the perturbation of nuclear translocation of IRF3 in response to Sendai virus infection, in cells ectopically expressing NopA.
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The stimulus for the album's idea was allegedly inspired by the reappearance of N. Senada, who supposedly provided the group with a tape containing samples of Eskimo sound samples and a jar of arctic air to record. The Harry Partch influence would once again come into play, as the language spoken on the album is entirely fabricated by themselves. As the story unfolds, recognisable words eventually become perceivable, when the story's leader becomes contaminated with North American pop culture.
Next, we investigated whether NopA binding to Ran can affect the Ran GDP/GTP ratio that is required to fuel nucleocytoplasmic transport. U2OS cells were challenged either with WT C. burnetii, the dotA :Tn, nopA :Tn, or the nopA :Tn complemented strains. Noninfected cells were used as control. Twenty-four hours postinfection, cells were lysed and incubated with agarose beads coated with the Ran effector Ran-binding protein 1 (RanBP1), to specifically pull down the GTP-bound form of Ran.
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For C. burnetii effector translocation assays, cells were cultured in black, clear-bottomed, 96-well plates and infected with the appropriate C. burnetii strain (multiplicity of infection [MOI] of 100) for 24 and 48 h. C. burnetii-expressing BLAM alone or BLAM-tagged CBU0021 were used as negative and positive controls, respectively. Cell monolayers were loaded with the fluorescent substrate CCF4/AM (LiveBLAzer-FRET B/G loading kit; Invitrogen) in a solution containing 20 mM Hepes, 15 mM probenecid (Sigma) pH 7/3, in Hank’s balanced salt solution (HBSS). Cells were incubated in the dark for 1 h at room temperature and imaged using an EVOS inverted fluorescence microscope. Images were acquired using DAPI and GFP filter cubes. The image analysis software CellProfiler was used to segment and count total cells and positive cells in the sample using the 520-nm and 450-nm emission channels, respectively, and to calculate the intensity of fluorescence in each channel. Following background fluorescence subtraction using negative control samples, the percentage of positive cells was then calculated and used to evaluate effector translocation. A threshold of 20% of positive cells was applied to determine efficient translocation of bacterial effector proteins.
Effector-coding genes were amplified from C. burnetii RSA439 NMII genomic DNA using primer pairs indicated in SI Appendix, Table S5. PCR products were cloned either into pLVX-mCherry-N2 or pRK5-HA plasmids to generate N-terminal-tagged mCherry or HA fusion versions of all effector proteins, respectively. For cloning of Ran in pcDNA3-GFP10-zipper, Ran was amplified using forward primers Ran-BspEI and reverse primers Ran-XbaI-rev.
2.c. Cheetahs: Genetic Drift and Loss of Diversity Caused by an Ancient Population Bottleneck
One of the best ways to bring down deaths is to diagnose cancer early, said Rebbeck. But even when these technologies are developed for widespread use they could exacerbate disparities for the poor, underserved and uninsured. That's because there remains systemic hurdles of access and cost.
To zero in on the gene behind this so-called "industrial melanism", Dr Saccheri's team started with traditional genetic mapping. They crossed black and pale moths in the lab and tracked which genetic "markers", dotted along each chromosome, seemed to be linked to the black colouring.
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Bodyguarding a Badass: Zigzagged. When confronted by would-be assassins in an alley, Steve's first instinct is to protect Diana. Once she proves fast enough to block bullets with her bracelets, she proceeds to protect him.
Unusually Uninteresting Sight: At the gala, Wonder Woman (disguised as an European woman) wears a sword in full sight down the back of her (backless) dress. No one ever seems to notice this, or to even look twice if they do.
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Ectopic expression of either mCherry-NopAFL or mCherry-NopAC-ter in combination with GFP-Ran in U2OS cells also confirmed the colocalization of both proteins at nucleolar structures labeled with the anti-fibrillarin antibody (SI Appendix, Fig. S3B), Conversely, Ran-GFP accumulation at nucleoli was lost when the small GTPase was ectopically expressed in U2OS cells in combination either with mCherry alone or mCherry-NopAN-ter (SI Appendix, Fig.
A live album of The 13th Anniversary Show: Live in Holland also gets released. During one Norweigan show that took place within the Arctic circle, the band would have to apply improv skills to their looks, and to their music (https://vgtdecor.ru/hack/?patch=6500). Their equipment never showed, so they had to borrow instruments from the audience (including ones they have never used nor seen before) and concoct makeshift costumes out of whatever they could find for the performance. An announcement was made that ticket holders could get refunds if the isea was too odd for them.
Many analysts imagine a new twilight struggle against the People’s Republic of China. More improbably, an equally dedicated band is treating the Russian Federation as America’s eternal enemy.
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The band appear at another film event, California's Mill Valley Film Festival, but, in person. They were honored with a 25th anniversary retrospective, and clips of various videos were shown as well as having been given a birthday cake.
The Residents had always planned to make a film, (something of which they always wanted to do before making music), and thus they began a rather ambitious and still as-of-yet unfinished self-financed project, Vileness Fats. The band's space at 20 Sycamore was converted to a soundstage, and 1/2" black-and-white video tape was used as the medium, which was a new technology available at the time. Not only was it suitable for it's cost effectiveness as compared to using film, but the band knew that it was also considered to be the 'next big thing'. The ongoing shoot was guided by a vague and unfinished script that included a cast of characters who lived in Vileness Flats, a community of one-armed midgets (including siamese twin who were tag-team wrestlers), who had to protect their town from the oncoming threat of the Atomic Shopping Carts.
Super Serum: Maru provides Ludendorff with a drug that gives him inhuman strength and stamina. The gas allows him to crush a pistol with his bare hands and momentarily trade blows with Diana.
In Project Altered Beast the story follows a man called Luke Custer who is a "Genome-Cyborg," a human whose DNA and other genetic make-up has been artificially altered allowing micro-chips containing the genetic make-up of other creatures to transform him into an anthropomorphic beast. After surviving a helicopter crash, Luke loses his memory and sets off to learn about the truth behind his past and the Genome-Cyborgs.
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Humans Are Special: Played with. Diana initially believes the best of humanity, as they were created by the gods, but this view is eventually corrupted by what she experiences in World War I.
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Heterochromia is the condition where a person has 2 different eye colors. These can either be 2 different colors in the same eye, or both eyes may be differently colored.
The BKWT channel and the BKG354S mutant channel display different selectivity. Raw current traces from BKWT (A), or BKG354S channels (B), from either 140 mM K+ or 140 mM Na+ in the internal (bath) solution. Currents were recorded from excised, inside-out membrane patches, with 140 mM K+ in the extracellular (pipette) solution. Currents were activated by a step to 200 mV, followed by voltage pulses ranging from 200 to −100 mV (100 to −20 mV for the mutant), in 10-mV decrements.
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A UK band by the name of Renaldo and the Loaf, (Brian Poole and Dave Janssen), big fans of the The Residents and signees to Ralph Records, drop by the Cryptic office to have them produce a music (https://vgtdecor.ru/hack/?patch=4060) video of the title track from their Songs for Swinging Larvae album. The album sold rather well for the label, and both bands would collaborate on a project that would end up being shelved indefinitely. Over the next while, the label would also sign other artists such as Yello, Fred Frith, Art Bears, DooDooettes, Half Japanese, Deficit Des Année Anterieurs, and Negativland.
Genetic disruption of ion channels underlies several neurological diseases, suggesting that ionic disturbances are common neuronal stressors potentially amenable to therapies. The detailed intracellular pathways coupling ion channel mutations to neuronal damage are largely unknown. Here, we describe the finding of a single loss-of-function mutation (https://vgtdecor.ru/hack/?patch=8214) in the BK channel in a young patient with progressive cerebellar degeneration. The mutant BK channel caused a profound dominant-negative effect on native channels, combined with reduced ion selectivity, leading to depolarization and depletion of mitochondria, and when delivered virally to mice, mimicked the disease. BK channel active drugs rescued the mutant cellular phenotype. These results point to the importance of mitochondrial ionic homeostasis in cerebellar disease and suggest therapeutic strategies.
Studies of a mouse model for another form of ataxia, SCA1, provided a possible explanation that incorporates an integrated model. In SCA1 mice, induced by genetic overexpression of the ataxin-1 protein bearing an expanded polyglutamine repeat, secondary loss of BK channel expression is accompanied by reduced spontaneous discharges. The discharges reappear during Purkinje cell shrinkage. The authors were able to rescue the cells from shrinkage by pharmacologic activation or overexpression of normal BK channels. They suggest that the shrinkage is a homeostatic adaptation to restore Purkinje cell firing in the absence of BK channels (52). However, again the viability of the mitochondria has not been evaluated in any of these mouse ataxia models.
In September, a film by the name of Conceiving Ada premieres at the Toronto International Film Festival. The film is a light science fiction time travel story incorporating real and CGI generated images that also includes a near-death Timothy Leary in a supporting role. The band provide the soundtrack, which was reported to be of an "electronic Victorian" genre.
Computer animation pioneer and illustrator Jim Ludtke, an Apple Computer affiliate, would create an innovative promotional 3D video for "Harry the Head". This would the group's first video in years, since they were upset with the fact that their visuals were too innovative for broadcast outlets such as MTV, which of course would create a roadblock in getting financing to create them.
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Long-Range Fighter: Merman, Garuda and Dragon forms are primarily suited for long range attacks. For the former two especially, the game turns into a SHMUP.
Because mice with targeted deletion of the BK channel α-subunit develop ataxia, we hypothesized that the LOF BK channel mutation (https://vgtdecor.ru/hack/?patch=1978) acts in a similar fashion. To test for a direct effect of BKG354S on cerebellar function, we engineered adenoassociated virus 9 (AAV9)-based expression vectors to deliver BKWT or mutant BKG354S channels to neonatal mice. We injected 2 μL at 1 × 1012 vg/μL of AAV9-BKWT or AAV9-BKG354S into mouse intracerebroventricularly at postnatal day 0 (P0) (33). Mice tolerated the injections well and developed normally with no visible neurological deficits initially. Analysis of the histological features of the cerebellar cortex shows no obvious difference between mice injected with BKG354S vs. BKWT channel, a finding similar to that of BK knockout mice.
Elemental Rivalry: Luke's basic form is the Werewolf, a canine beastman with a light blueish aura. His main opponent and rival, Brad, possess the power of the Weretiger, a feline beastman with a dark reddish aura. According to The Other Wiki, the Werewolf is light-elemental, while the Weretiger is dark-elemental.
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Try to Fit THAT on a Business Card: "Diana of Themyscira, Daughter of Hippolyta, Queen of the Amazons". If given half a chance, Diana will introduce herself by rattling out the whole thing.
A study published in 1987 compared genetic diversity in cheetahs to genetic diversity in other cat species, and in three non-cat species (click here for an article in the NY Times about this study. Click here for a more in-depth article from Scientific American. For the original study, click here).
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Complete heterochromia is very rare in humans. The other 2 forms, however, have a higher frequency than the first one. Furthermore, humans are not the only ones to have this variation.
Forty songs/forty minutes= 1 minute songs. The minute (pun intended) tracks resembled that of radio spots (commercials). The synthesizer plays a heavy role throughout this masterpiece; prickling, prodding and tingling the listener's blood pressure with the sounds of truly enjoyable elevator music. However, we only hear what it leaves in it's wake as it passes through the elevator shaft- a whirling, resonating doppler effect that somehow had managed to stop by a theatre to watch a couple of B movies while en route to your ears. Several contributors are listed in the album credits- Snakefinger (of course), Fred Frith (who would soon sign a recording contract with Ralph Records) and XTC's Andy Partridge.
Duel Boss: 3 boss fights occur without any mook harassing you: Weretiger, Rhinorian (it even will sit still and recover its health slowly if you turn into human form), and Anastasia. Instead of taking Spirit Gauge from enemies, you can find them being dropped around in Weretiger's room; you can use your human form's Deathblow attack on Rhinorian (although it'll only make the boss drop Spirit Gauge droplets instead of you absorbing it directly); and attacking Anastasia enough times will make her drop the droplets.
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True Blue Femininity: Diana wears a blue gown when she infiltrates a gala to kill Ludendorff. She stole it from a German noblewoman.
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The new millennium brings Icky Flix- the band's first foray into the world of DVD. The band's entire set of music videos were included, (by now their Third Reich N' Roll video had been inducted into New York's Museum Of Modern Art) as was restructured ones, newly created pieces, and excerpts from Vileness Fats.
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Animorphism: The main mechanic via Spirit Orbs. You get them by beating up flashing enemies and picking the Spirit Orb that falls out.
Garuda's Tornado is a Tornado Move that damages all around you. If upgraded, you can move around while the Tornado is active, making it able to hurt more.
When the team gets pinned down by a German sniper hiding in the church tower, it seems like this will happen with the team's sniper Charlie. Unfortunately, Charlie freezes up from PTSD and is unable to take the shot. Fortunately, Steve remembers an Amazon using a shield as a platform to leap behind some Germans, and has Sameer and Chief lift a large metal panel for Diana to leap off it into the tower.
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Unlike in typical elections, there is no primary to winnow the field. A low bar to entry all but ensures that scores of candidates will surge into the race. As in 2003, the allure of being governor is likely to draw in not just politicians but affluent business executives or celebrities — some of whom will bring their considerable resources to bear.
Baleful Polymorph: The cutscenes show Neff turning Athena into a bird. Once he's dead, she's returned to normal.
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However, the bacterial effector/s involved in this process remains uncharacterized (9). We have previously reported the large-scale phenotypic characterization of the C. burnetii transposon mutant library, which led to gaining important insights into the function of the Dot/Icm secretion system, and which highlight an important set of virulence determinants (10⇓–12). Importantly, several genes involved in intracellular replication of C. burnetii encode proteins with predicted eukaryotic-like domains, which prompted us to investigate eukaryotic-like genes (EUGENs) on a genome-wide scale. Here, we identify and validate the Dot/Icm-mediated translocation of seven C. burnetii EUGENs. Among these, NopA (nucleolar protein A) displays four regulation of chromosome condensation (RCC) repeats, which are partially homologous to the seven repeats found in the bladed β-propeller structure of the Ran GEF RCC1 (13⇓–15). Similarly to RCC1, NopA also localizes at the nucleus of infected or transfected cells; it is found associated with the chromatin nuclear fraction; and it uses the RCC-like domain to interact with Ran. Differently from RCC1, however, NopA accumulates at nucleoli and sequesters Ran, thus perturbing nucleocytoplasmic transport. Indeed, NopA perturbs nuclear translocation of p65 upon cell treatment with TNF-α or challenge with C. burnetii. Conversely, transposon insertions in the nopA gene restore nuclear translocation of p65 during infections, to levels that are similar to those observed with the Dot/Icm-deficient C. burnetii dotA mutant. Accordingly, myeloid cells challenged with the C. burnetii nopA or dotA mutant strains present a functional innate immune response, as opposed to myeloid cells exposed to wild-type (WT) C. burnetii or the nopA complemented strain.
Finally Eskimo, an impressionistically ambient concept album is released. It is intended to be heard in its entirety, and it tells a self-invented story of the Eskimo people who inhabit the North Pole region, and done so solely with sound. A written explanation of the story is available in the liner notes, and meant to be read while listening to the album with headphones.
The pattern of heterochromia, or the uneven distribution of melanin, can occur in 3 different ways. Thankfully, no matter how uncommon heterochromia is, there are people that save us the burden of having to imagine such a beautiful occurrence!
After 24 h of infection, cells were washed in PBS and pelleted at 4 °C. Transfected or infected cell pellets were subjected to cell fractionation as previously described (35). Where appropriate, cytoplasmic, nuclear, and chromatin fractions were subjected to immunoprecipitation using 40 μL of anti-HA magnetic beads (Sigma) for 2 h at 4 °C with rotation. Bound proteins were eluted using 80 μL of 100 μg/mL−1 HA-peptide (Sigma) and then resuspended in Laemmli buffer 4× and analyzed by Western blot.
The film's context and background is implied to be the time of Erich Ludendorff's Spring Offensive, Imperial Germany's last smash-and-grab to defeat and conquer the Western Front, making their greatest advances. The film implies that the armistice and peace negotiations are what is leading to the end of the war, rather than Ferdinand Foch's Hundred Days Campaign, the great Allied counteroffensive that brought them to the gates of Germany and actually caused them to surrender, alongside the Revolution in Russia and its spread into Germany.
Darker and Edgier: Compared to previous Altered Beast games, this reboot is a Deconstruction on those games putting into a Film Noir Crapsack World setting where Body Horror are much more visible. The main characters are much more immoral and borders into Sociopathic Hero territory, among others.
Bonus Dungeon: The Elevator of Doom is an extra game you can access after beating the story, where you'll be pitted against waves of enemies in a large elevator. At first you have 4 elevators, each forcing you to use 4 of your basic forms; completing those four will unlock 2 elevators that forces you to use your 2 other forms; completing those two will unlock an elevator where you're free to use any form, and completing that one will unlock another multi-form elevator that leads to the Bonus Boss.
Over the past few weeks, we have shown you a massive project by Bill Gates that is so all-encompassing that it quite nearly beggars belief. That expression is an Old English term which means something that is so unbelievable that it nearly cannot be believed. The Nazi concentration camps in the Holocaust are a great example of that. When the films of what went on there came to light it was so astonishing that it was hard to comprehend such a thing could happen.
Taken for Granite: One of Werebear's powers — the Bear Breath — enables it to turn enemies to stone by breathing on them for a One-Hit Kill. However, it doesn't work on Mouldy Snail or any of the other bosses.
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Dying Smirk: Steve has to mentally psyche himself up in order to blow up the plane carrying Doctor Poison's deadly gas and foil the villains' evil plan. He is visibly afraid and clearly doesn't want to die, but the moment before he shoots one of the gas canisters, he does smile.
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Bigfoot, Sasquatch and Yeti: The third boss is a Sasquatch. Played with in that it's a mutated (https://vgtdecor.ru/hack/?patch=5311) frog with a body resembling a gorilla. The Wendigo form you get from it resembles a sasquatch or yeti better.
Although the association of other LOF K+ channel mutations (https://vgtdecor.ru/hack/?patch=7490) in the ataxia (1, 3, 6) suggests that a persistent depolarized neuronal state is a common pathological pathway, determining the subcellular sites of action of the toxic effect of these mutations will provide significant insight into the pathogenesis of SCAs. BK channels are widely expressed throughout the central nervous system and are abundant in cerebellar Purkinje cells. Together with SK channels, they play a key role in quickly repolarizing the cell membrane when the intracellular Ca2+ concentration is increased, as well as in shaping the afterhyperpolarization potential, especially in mature Purkinje cells (45, 46). Both BK and SK channels are also present in membranes of organelles, for example in the inner mitochondrial membrane (47) and in the inner nuclear membrane (48, 49), where they have similar biophysical and pharmacological properties. In mitochondria, they play a role in calcium retention capacity coupled to the permeability transition pore.
Dragon has Thunder Cannon, a Wave-Motion Gun that serves as an alternative to its Limit Break. Dragon's own normal attacks are also charged attacks, where holding the Square button will cause it to create more and more electric balls in its hands (depending on how far you upgraded its combo) to be thrown towards enemies.
On the contrary, the expression of GFP1-9 in combination with GFP10-Ran and GFP11-RCC1 allowed reconstitution of GFP fluorescence, which was homogeneously detected in the nucleus in over 73% of transfected cells (SI Appendix, Fig. S3 A and D). Lack of GFP reconstitution upon expression of GFP1-9 in combination with GFP10-Ran and GFP11-fibrillarin indicated that the shared nucleolar localization was not sufficient for GFP reconstitution (SI Appendix, Fig.
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You Don't Look Like You: An action figure and LEGO set both show Ares sporting a helmet that resembles a ram's skull and a face that's blackened out except for his eyes. In the film itself, Ares wears a different kind of helmet and his face looks completely human. Also, he's of average height, whereas the LEGO set showed him being much bigger than a person.
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For nopA complementation, the nopA putative promoter and nopA sequences were amplified using the primer pairs NheI-prom1217-fw/PstI-prom1217-Rev and CBU1217-BamHI/CBU1217-EcoRI-rev, respectively (SI Appendix, Table S5). The PCR products were cloned into pUCR6K-miniTn7-Kan-tetRA-4HA. Plasmids were electroporated in the nopA :Tn mutant strain Tn227 (10).
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The 13th Anniversary Tour concluded with a show at the beginning of the year with Penn and Teller as guests. The infamously silent Teller promised to sing a Residents song in return that one of the Residents revealed themselves in front of the audience. The exchange occurs, and the Resident takes off his/her eyeball to reveal Teller underneath.
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In the final battle, killing Ares doesn't automatically stop the war. Diana has to face the fact that sometimes humans just fight among themselves for their own reasons and horrible things happen in war without the machinations of a Big Bad, especially when one side is led by people like Ludendorff.
Spiders Are Scary: Xenesthis is one of the biggest bosses, overall realistic-looking and one of his attacks consist in puking out his previous meals at you. The fact that you have to get as close as possible to it to make it pounce at you (so that he becomes open after that) doesn't help, nor the mention that its spiked forelimbs are used to hold the prey still as it eats them.
Improbable Aiming Skills: One of the German soldiers who landed on Themyscira managed to shoot a bullet perfectly into a moving target: an Amazon fast-swinging down from the cliff at 45° angle, at long distance, on first shot. You gotta admit, the guy's skilled. So much so that you might think it was a stray bullet, or at least aimed at Steve or Diana, who dodge.
Opinion makers do that (and not for Anti-Asian crimes like this one, either). I will skip the moral panic over hatred for Asians, which conveniently erases the deaths of two white women, and the injury of a Hispanic man.
Having existed for over twenty years, the group would develop a Pink Floyd: The Wall type syndrome, feeling as if they were seen as nothing but freaks on stage, instead of the musical (click to read more) artists that they were. Suffering with this oppressive angst, the band decide to transform the concept into a creative one, and the Freak Show album would surface. Characters like "Harry the Head", "Wanda the Worm Woman" and "Jello Jack the Boneless Boy" would all be given their own track, each exploring a facet of how people are treated in the world. It would also become their most ambitious project to date, which would evolve and mutate (you could try here) into many other mediums. Fox recounts the album's importance; "Compositionally, most music (https://vgtdecor.ru/hack/?patch=6733) tends to have things that repeat- some element in it that reoccurs, like hooks choruses and verses. It isn't completely without that, but for the most part, it is stream-of-consciousness. Compositionally, it goes from point A to point Z, and doesn't come around to try to hook you with any reoccurring melodies.
We Can Rule Together: Ares tries to convince Diana that they're kindred spirits and can revert the world to a paradise once humans are eliminated, trying to show her that Humans Are the Real Monsters. It doesn't work, because she knows at least one who isn't.
Diana is left in such a daze after watching Steve's Heroic Sacrifice, teetering dangerously close to the Despair Event Horizon, and then vanquishing Ares. She's just emotionally spent after all that.
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Villain-Beating Artifact: On the hidden island of Themyscira, the Amazons have been guarding an ancient sword known as the God-Killer. It's the only weapon capable of killing Ares, the evil deity responsible for spreading death and destruction among humans and the one who wiped out the rest of the Greek pantheon when they wouldn't side with him. Subverted when Ares reveals it to simply be an ordinary blade. The actual God-Killer is Wonder Woman herself, who is Ares's sister through their father Zeus. Gods can only be killed by other gods.
After Ares's defeat, several German troops remove their gas masks to reveal themselves as boys in their mid-teens. By this point in the war, this practice was sadly common to both sides, as entire generations of older men were destroyed in previous battles. Germany especially needed Cannon Fodder since it didn't benefit from fresh troops like the Allies when the USA joined in the war in 1917, and thus had to scrape the bottom of the barrel.
In February, File, a Canadian art magazine, included a flexi-disc sampler of Meet The Residents with a absurdist advertisement promising a full copy of the album in exchange for only two dollars. They sold 40 more copies as a result.
Continuity Nod: Diana in Batman v Superman: Dawn of Justice was looking for a valuable picture, revealed as a picture taken of her in WWI, but only a digital scan of the original picture (along with other secret files on her). This film shows the physical picture and the story about how it was taken.
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The difference in costs often boils down to domestic state capacity: bureaucracies in East Asia and Continental Europe tend to be better-staffed and more empowered to make professional decisions. The details are naturally more complicated, but the pattern is nonetheless clear: the countries with the lowest infrastructure costs are also the countries where the state acts swiftly, with mechanisms that limit the lag between financing and construction.
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Arrogant Kung-Fu Guy: Brad, a cold and haughty guy who was the first Genome Cyborg, and a rather powerful one too; he was the one wounding Luke in the past, and it shows more as a boss - he can Flash Step out of your attacks and slug you just as quickly. He even would take time to cross his arms and tap the ground in boredom or Finger Wag you, should you get knocked down; he's also open to attacks during such.
There are well-documented agglomeration effects of the innovation economy—you want proximity to the other engineers and programmers and, not to mention, the venture capitalists—but the bigger reason for Big Tech’s role in regional inequality is more about economic policy. So much of our geographic concentration is tied to market concentration. The business, commerce, and wealth that used to be spread all across the country, in various industries, are now increasingly dominated by a few companies that reside in certain places. It happens with media ad revenue, which used to be spread everywhere among newspapers and local TV and radio but is now increasingly hoovered up in the form of digital ad revenues to the two companies [Facebook and Google] that control 60 percent of that market, that both reside in the Bay Area. And retail that used to be spread all around the country, from mom-and-pops up to regional department stores, is now increasingly dominated by a company that resides in Seattle. And so, that was how I arrived at Amazon.
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Given the role of Ran in nucleocytoplasmic traffic, and the previously reported observation that C. burnetii infections modulate nuclear translocation of p65 by a Dot/Icm-dependent mechanism (9), we investigated whether NopA affects the nuclear localization of p65, which follows the activation of the NF-κB signaling pathway. U2OS cells transfected with plasmids encoding either HA- or mCherry-tagged versions of NopA were either left untreated or challenged with 10 ng/mL TNF-α for 30 min, and the nuclear translocation of p65 was monitored using an anti-p65 antibody either by fluorescence microscopy or Western blot following cell fractionation. Cells expressing either HA- or mCherry-tagged CvpB or the tags alone were used as controls.
The group back up the equally strange entity known as Schwump, a Portland, OR radio personality for his "Aphids In The Hall" single. Around this time period N. Senada reportedly vanishes.
New Game+: The game throws out a very subtle one. If you manage to acquire Weretiger - meaning that you have completed the game 100% - you can start a new game with all of your beast forms unlocked from the start.
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Mana Drain: In your human form, your "Deathblow" move can siphon Spirit Gauge from enemies. If the enemy is red colored (which will drop life essences if killed normally), using Deathblow on them results in Life Drain instead.
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Nintendo Hard: The arcade version and its ports. Especially the ports, which give only three lives, no healing items, no extra lives, no Mercy Invincibility, and no continues.
U2OS were grown in Dulbecco’s modified Eagle medium (DMEM) supplemented in 10% (vol/vol) fetal calf serum (FCS) at 37 °C and 5% CO2. For the interaction assay, U2OS cells were cotransfected with Lipofectamine 2000 (Gibco, Invitrogen) with plasmids encoding for GFP1-9, GFP10, and GFP11 fusions. At 24 h posttransfection, cells were fixed in 4% paraformaldehyde in PBS solution and processed for immunofluorescence. Protein/protein interactions were scored by calculating the percentage of GFP-positive cells over the total number of cells positive for the anti-GFP antibody.
Deliberate Values Dissonance: Downplayed, but it still comes up. Diana gets pretty quickly kicked out of a military summit in London for being a woman, and is treated condescendingly by Steve Trevor's superiors. Etta Candy offhandedly mentions the fact that women haven't been given the right to vote in the United Kingdom yet in a separate scene. Sameer talks about wishing to become an actor but was denied the opportunity because of the color of his skin. Chief mentions how the Native Americans were wiped out and displaced by white Americans prior to the start of World War I. And if anybody is homosexual, they're certainly not going to make it obvious.
The Germans have no way of stopping Diana once she actually steps into the war, and this is a long time before Superman would make himself known, so they have zero idea of who or what they're dealing with. After they've been holding off the Allies for four years, it takes Diana about 15 minutes to break through their defenses and liberate a town. The only thing they've got that can do anything to slow her down is a drugged-up Ludendorff, who doesn't last all that long despite his newfound strength.
Welcome to Now The End Begins, and I think now you see why we say it’s the ‘front lines of the end times’. Flight #777 preparing for takeoff.
We Are as Mayflies: Steve and Diana talk about his father's watch and Diana seems confused over why he would lead his life according to such a thing. Being an immortal, she doesn't have the same perception of time. As Steve races off to destroy the bombs, knowing he won't return, he tells her that he wished they had more time together. At this point, she starts to understand.
Diana finds her own outfit to get into the gala by coming across an obnoxious socialite in a blue dress; amusingly, all we see Diana do is size her up and compare their heights while the lady is confused. In the next cut, we see Diana wearing it.
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I think not, but it may take a generation or two for the experiment to fail. Which is, no doubt, why social movements like Black Lives Matter are so beloved at, for example, Davos.
Once More, with Clarity: When Diana and Steve speak to each other for the very last time, the sound of their voices is muffled and indistinct, almost as though Diana's hearing is still recovering from the explosion that had just knocked her onto the airfield. However, when Diana later recalls the conversation, the scene is played again, this time with the vocals played clearly. It turns out that Steve had decided to give his life to blow up the poison gas bomber, and he was letting Diana know that this was their final goodbye. That memory inspires Diana to spare Dr. Maru and reject Ares once and for all.
The localization of CBU1217 was further investigated by cloning the gene into a pJA-LacO-4HA plasmid, to express the effector protein carrying an N-terminal 4×HA tag in C. burnetii, under the control of an isopropyl β-d-1-thiogalactopyranoside (IPTG) promoter, and monitor its localization during infection. GFP-expressing WT C. burnetii or the dot/icm mutant dotA :Tn (10) were transformed either with pJA-LacO-4HA or with pJA-LacO-4HA-cbu1217. Expression of 4HA-CBU1217 was validated by Western blot using an anti-HA antibody (SI Appendix, Fig.
He funds Steve's mission because it will require Diana to experience the Western Front first-hand. At the point that Sir Patrick calls off the mission, every possible outcome will result in Diana being disillusioned. If she goes ahead and kills Ludendorff, she'll be horrified that the war doesn't magically end. If she goes back to London, she won't reach the city until its millions of inhabitants have been gassed. If she stays in the town she liberated, she'll have a front-row seat to it and her friends being gassed. Every outcome leads to Diana's conception of humanity being inherently good stripped away. What Ares can't conceive, however, is that Diana won't share his belief that man deserves to be exterminated, that she could still see their better nature while being forced to grow aware of their worst nature.
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Exports, however, are having the worst year since 2007. On top of a trade war and the world pandemic, import container counts continue to surge. There is chaos in container movements with containers in the wrong place and shortages of rail cars to move containers – however, the container situation again improved this month – but there continues a shortage of containers and unloading berths.
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A Lizard Named "Liz": There are two bosses named Xenesthis and Callichthys. Their names actually tell you that they're a Giant Spider and a giant catfish respectively.
For coimmunoprecipitation experiments, pLVX-GFP-N2-tagged WT Ran, RanT24N/Q69L/N122I mutants, or vector control were cotransfected with pRK5-HA-NopAC-ter in U2OS cells. At 24 h posttransfection, cells were lysed in lysis buffer (10 mM Tris HCl, pH 7/5, 150 mM NaCl, 0/5 mM ethylenediaminetetraacetic acid [EDTA] 1% Nonidet P-40) supplemented with a protease inhibitor tablet (Complete; Roche) and incubated with 25 μL of GFP-Trap magnetic beads (Chromotek) for 2 h at 4 °C with rotation. The beads were then washed three times with wash buffer (10 mM Tris HCl, pH 7/5, 150 mM NaCl, 0/5 mM EDTA), resuspended in Laemmli buffer 4×, and analyzed by Western blot.
Similarly to the eukaryotic protein RCC1, NopA interacts with Ran, with preferential affinity for the GDP-bound form and promotes the activation of Ran. Differently from RCC1, however, NopA also triggers a nucleolar accumulation of Ran. Thus, the observed increase in the intracellular levels of GTP-bound Ran may result from either GEF activity of NopA (which has been reported for RCC1), or via the observed sequestration of Ran at nucleoli, which would prevent GTP-bound Ran to recycle back to the cytoplasm, where Ran GTPase-activating proteins (GAPs) stimulate GTP to GDP conversion. As we were unable to purify sufficient amounts of either full-length NopA or NopAC-ter, we could not assess for the moment whether NopA has intrinsic GEF activity. Interestingly, a residual increase in the intracellular levels of Ran-GTP was still observed in cells challenged with the nopA :Tn mutant strain, as compared to infections with the Dot/Icm-defective dotA :Tn strain. This may suggest that other C. burnetii effectors may have a role in the modulation of Ran activity.
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Archaic Weapon for an Advanced Age: Wonder Woman fights with a sword, shield, and lasso against WWI Germans using contemporary weapons and tanks. She's shown using her shield to protect herself from machine gun fire and even deflect mortar shells.
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Finally, when he's stuck on the roof and Diana reveals her origins and her intention to destroy Ares, he clearly has no idea what Diana is talking about. These clues are clear signs that show Ludendorff isn't really Ares.
Science Fiction Dictionary: Ideas, Terms and Technology
Death of the Old Gods: All of the Greek Pantheon sans Ares and Diana are dead. By the end, only Diana is left, which leaves room for the incoming New Gods of Justice League (2021).
Stone Wall: The Minotaur is slow but has the greatest defense of all the transformations. When other beastmen are knocked back by an attack they fall on their back, even the larger Wendigo, but the Minotaur instead is merely knocked back on his knee, even from such powerful attacks. His Iron Guard even allows him to stop giant bosses and advancing wall/rolling rocks traps dead in their tracks.
The Amazons as a whole are reasonable when Diana rescues Steve from drowning, which leads to the Germans invading and opening fire on the Amazons, who were only holding up a defense. When several suggest killing Steve since he brought invaders to their paradise, Antiope's comrades point out that he's the only one who can answer their questions. After they interrogate him using the lasso, they dress his wounds and offer Sacred Hospitality while debating whether or not to set him free. Steve is grateful to be alive, and tries to get some relief from his Gilded Cage.
Shapeshifting Heals Wounds: Lorewise, at least, Luke can technically regenerate from injuries as he transforms into one of his beast forms. In a subversion, he has a Wound That Will Not Heal on his chest, courtesy of his rival, Brad, whose Weretiger claws are laced with a certain virus that causes cellular necrosis; Luke retains the scar even in his beast forms. In gameplay, he retains his current HP whenever he transforms.
Notice that the “class structure” (as determined by ownership of capital vs. selling your labor power to survive) on the horizontal plane is identity pre- and post-equity; 90% of all capital is held at the tippy-tippy-top. Only the proportion of colors — pink, black, and yellow — changes at each level.
Early in the movie, General Ludendorff visits Dr. Poison to check on the progress of the gas weapon she developed, but she insisted that she can't improve it further without her journal that Steve has stolen from her. A gust of wind then blows one of the pieces of paper notes she had thrown away back to her. After she reads it, she instantly realizes what she needs to do. Also, just a scene earlier, she gives Ludendorff a gas pill that gives him superhuman strength, telling him that she came up with it the night before. In the climax, it is revealed that these are the works of Ares, who indirectly provides Dr. Poison with new ideas of deadly weapons.
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We also analyzed the kinetics of the K+ current activation. The currents from BKG354S were activated up to two orders of magnitude slower than the currents from BKWT. Interestingly, the time constants from BKWT-G354S were only two to six times slower than the currents from BKWT (SI Appendix, Fig.
The Pennsylvania Amish were founded by about 200 German immigrants in the mid 1700s. The allele for Ellis-van Creveld syndrome has been traced to one couple from the founding population. And because the Amish tend to marry only within their own culture, the chance of two Amish being heterozygous is much higher than it would be if they married with the surrounding non-Amish population.
Mice with targeted deletion of BK channel α-subunit (BK−/−) have predominantly a cerebellar phenotype with intention tremor, gait incoordination, and impaired eye-blink conditioning (a classic form of cerebellar motor learning). Surprisingly, these animals show no cerebellar degeneration. We found a similar picture in a mouse model induced by AAV delivery of the BKG354S mutation. This is in contrast to two siblings with the more widespread motor system disorder and marked cerebellar atrophy, attributed to homozygous LOF mutations (read this article) in the BK channel. We speculate that the difference relates to age differences, or the role of compensatory pathways. The present patient has a milder phenotype in which progressive ataxia began at age 8 y, attributed to a BK channel LOF mutation.
Whereas the population bottleneck that Northern Elephant Seals suffered was observed in recent history, we can infer that cheetahs suffered a similar bottleneck at some time in their past. The evidence is in the cheetah’s gene pool.
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Kill the God: The ultimate purpose of the aptly named "God-Killer" is to slay Ares, who himself had killed the rest of the Greek Pantheon. Diana was told it was a sword, which she stole to fulfill its supposed purpose, but as Ares states, only a god can kill another god — the actual "God-Killer" is Diana herself, as a child of Zeus.
All Your Powers Combined: Anastasia can do many things Luke can do with his beast forms. As mentioned by her, both are chimera-type Genome Cyborgs (while Brad isn't).
2b. Elephant Seals: Genetic Drift and Loss of Diversity Caused by a Recent Population Bottleneck
As he tells Diana, he really didn't have to corrupt men; humans did that on their own. He wants to help wipe them out so that the world won't be destroyed, and its former glory can be restored. During their fight, he starts on the defensive while trying to talk Diana down.
Tough Love: Why Antiope disregarded her sister's orders and started training Diana in secret — she had no desire for Diana to fight any more than Hippolyta did, but she knew that Ares was going to return one day and come after Diana, regardless of what any of them wanted. The only way to protect her niece was to make sure she could protect herself. Once Antiope sways Hippolyta to her way of thinking, the Queen of the Amazons makes sure her daughter is trained harder than any other Amazon before her. Antiope not only gained the permission to train her, but the edict to push Diana to her maximum potential; Hippolyta wouldn't settle for anything less than Diana surpassing Antiope herself.
The NopA/Ran interaction was further investigated in U2OS cells, following the ectopic expression of either HA-tagged NopAN-ter, NopAC-ter, or the C. burnetii effector protein CvpF as negative control (22). Unfortunately, under these conditions, we were unable to immunoprecipitate full-length NopA from transfected cells. Similarly to infected cells, 24 h posttransfection, cells were lysed, separated into cytoplasmic, nuclear, and chromatin fractions, and NopA truncations and CvpF were immunocaptured from cell fractions using an anti-HA antibody.
Aside from the above, there's also the Cockatrice and its King Mook Basilisk, a secret project of Dr. Jobs where he aimed to recreate an ancient animal; it has features of both bird and reptile. The Unique Enemies are variants of these with different properties.
Guys Smash, Girls Shoot: Inverted. Diana is a Flying Brick and is thus more likely to rush head-on into combat, while Steve and his companions tend to cover her through shooting guns.
Detectable BK labeling can be seen in PC12 pCDNA3-only transfection because of possible endogenous expression. After 24 h in the presence of NGF, no neurites are present in cells expressing the mutant, although a systematic check shows no difference between cells expressing BKWT or BKG354S.
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Neutral Female: Gender-inverted. Several German spies corner Steve and Diana in an alley, but Diana beats them up while Steve simply watches. It's most likely due to shock, as this is the first time he's seen Diana fight and she's casually deflecting bullets with her bracelets. Alternatively, he's smart enough not to get in the way of someone much better at this than he is.
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With intent to cash in on their new-found popularity, the Cryptics re-released the "Satisfaction" single, which ended up landing in the #1 position of Record World's New Wave charts, surpassing Devo's cover version of the same song. The sudden influx of attention makes them uneasy during the recording of their next album; fearing that their creative process would be might influentially polluted, so, the band flees to England with the tapes to continue the project-in-progress. While ransacking the vaults in hopes of finding something to release until the band would return with the new album, The Cryptics would find Not Available, and unleash it unto the public.
The 1918 group photo functions as one, as it triggers her memories and the people who appeared on it are long dead. And because it commemorated their rescue of a village that was gassed to death soon afterwards.
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Tactical Suicide Boss: If you run out of cars to throw at Aeshma he'll pull several out of the lot behind him and sending them sliding towards you. Xenesthis will try to pounce you if you get too close to the web, allowing you to whale on it when it climbs back on the arena.
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Shapeshifter Baggage: Luke can easily turn into monsters with elongated/enlarged limbs and bodies and, in case of Dragon, growing wings and a tail. Anastasia also turns gargantuan after activating her Chimera form.
Zack Snyder states she's 5000 years old as of Batman v Superman: Dawn of Justice. In a Collider interview, though, Patty Jenkins says that Diana is about 800 years old in World War 1.
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Weretiger has Genocide Rage, a rushing-forward attack similar to Werewolf's Berserker Charge but covers much longer distance and cannot be steered. Its own jump technique can also be charged like Werewolf's to make you jump higher.
Diana is disgusted when she learns that in modern warfare, generals don't lead from the front anymore, but even the "good" Allied side is led by men like Field Marshal Douglas Haig sitting safely in London while sending thousands of their own men to die in trench warfare. When she protests that their own soldiers are dying on the front every day the war is prolonged, Haig flippantly tells her "That's what soldiers are for".
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Also, she saves a village caught in the crossfire between the British and Germans; Charlie starts to sing again, as a result. Dr. Poison gasses the village as a demonstration.
Homing Projectile: Garuda can shoot homing Feather Flechettes while being stationary (if you're moving around, you'll fire auto-aimed feathers instead). Dragon can fire homing electric balls.
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Dragon's Arc Lightning is a burst of electricity all around you, capable of killing all but the strongest Mooks in one hit. If upgraded, its range is increased.
Dictionary of Science Fiction
Costume-Test Montage: First thing to do in London: get new clothes for Diana, with Etta Candy as guide. Most outfits are rejected for being impossible to fight in, or being itchy.
The entry of James McAvoy in X-Men: First Class was quite an interesting scene. He was shown in a bar, talking to a pretty, blonde lady. The most striking part about her appearance was her eyes – one was green and the other was blue. In fact, Charles Xavier commented upon her eyes as a conversation starter!
Directed Ty Roberts and financed by NEC, the band would perform Freak Show material which was recorded and then mixed live onstage with the software by multimedia experimentalist, record producer and musician, Todd Rundgren. Apple Computer would later incorporate this footage into their groundbreaking yet still infantile Quicktime software bundles.
It comes from 2 words; hetero – which means different, and chromia – which refers to color. It is the condition where a person’s eyes are of 2 different colors.
Take Up My Sword: Diana's tiara actually belongs to Antiope, her aunt and combat instructor who is killed protecting Diana during the beach battle. As she leaves the island, Hippolyta gives it to Diana to remind her of the greatest Amazon warrior who ever lived.
A full assessment of the probable heterotetramerization of BKWT and BKG354S subunits in the same channel is beyond the scope of this study. In our preliminary results, unitary currents studied by both single-channel patch clamp and by noise analysis of macroscopic currents in membranes from oocytes injected previously with cRNA for both types of proteins, never showed intermediary conductance. On the other hand, macroscopic K+ currents recorded from these cells showed evidence of heterotetramerization. First, BKWT expression, as measured by the amplitude of the macroscopic K+ currents, was affected by the likely coexpression, in the same oocyte, of BKG354S, suggesting dominant-negative effect and in agreement with the mitochondrial studies. Second, the time constants of the putative BKWT-G354S activation kinetics sit in between the BKWT and BKG354S values. Third, the voltage dependence of the activation kinetics resembles that of pure BKG354S channels: They are virtually voltage independent (SI Appendix, Fig.
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Villainous Friendship: General Ludendorff and Dr. Poison are genuinely loyal and friendly to each other. Ludendorff doesn't exert his Bad Boss tendencies on her, and Maru concocted a special Super Serum pick-me-up just for him.
Sexy Discretion Shot: After liberating Veld, Steve and Diana share an intimate moment together. Following their dance in the snow, Steve escorts Diana to her hotel room. Rather than leave, Steve enters too after Diana gives him an inviting look. They kiss passionately before the camera cuts away to the outside of the building, where the light in Diana's room is the only one still on.
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During the final battle, Wonder Woman gets a Heroic Second Wind against the villain and makes a renewed attack. The villain seems to be hard-pressed, but very quickly regains control and slaps her down with little effort, as he's still plainly much more powerful than her. She has to redirect his own lightning to kill him.
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The lasso doesn't cause Ares pain because he is Brutally Honest. He even turns the tables on Diana by using it to show her visions of the true world he desires.
It contains the musical (https://vgtdecor.ru/hack/?patch=2144) foundations of what the Residents would become, except in an analogous and primitive fashion. It was mutilated novelty nightmare jazz pop at its finest, making the musique concrete of Varese and Zappa feel like melted plastic. And, it was the first of its kind.
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The Stinger: Found only on the home-video releases. Etta (Lucy Davis) rounds up the boys for a mission after the end of the war; a mysterious artifact found in Liège and they are to recover and deliver it to the Americans — it's one of the Mother Boxes, the one used to create Cyborg decades later. It's a Sequel Hook for Justice League, in which the New God Steppenwolf goes on a rampage looking for them.
Luciferase is a generic term for the class of oxidative enzymes that produce bioluminescence, and is usually distinguished from a photoprotein. The name was first used by Raphaël Dubois who invented the words luciferin and luciferase, for the substrate and enzyme, respectively. Both words are derived from the Latin word lucifer – meaning lightbringer.