title: Bioluminescence

text: In a general sense, the principal chemical reaction in bioluminescence involves a light-emitting molecule and an enzyme, generally called luciferin and luciferase, respectively. Because these are generic names, luciferins and luciferases are often distinguished by the species or group, e.g. firefly luciferin. In all characterized cases, the enzyme catalyzes the oxidation of the luciferin.


title: Bioluminescence

text: In some species, the luciferase requires other cofactors, such as calcium or magnesium ions, and sometimes also the energy-carrying molecule adenosine triphosphate (ATP). In evolution, luciferins vary little: one in particular, coelenterazine, is found in 11 different animal phyla, though in some of these, the animals obtain it through their diet. Conversely, luciferases vary widely between different species, which is evidence that bioluminescence has arisen over 40 times in evolutionary history.


title: Bioluminescence

text: Both Aristotle and Pliny the Elder mentioned that damp wood sometimes gives off a glow. Many centuries later Robert Boyle showed that oxygen was involved in the process, in both wood and glowworms. It was not until the late nineteenth century that bioluminescence was properly investigated. The phenomenon is widely distributed among animal groups, especially in marine environments. On land it occurs in fungi, bacteria and some groups of invertebrates, including insects.


title: Bioluminescence

text: The uses of bioluminescence by animals include counterillumination camouflage, mimicry of other animals, for example to lure prey, and signaling to other individuals of the same species, such as to attract mates. In the laboratory, luciferase-based systems are used in genetic engineering and biomedical research. Researchers are also investigating the possibility of using bioluminescent systems for street and decorative lighting, and a bioluminescent plant has been created.


title: Bioluminescence

text: Before the development of the safety lamp for use in coal mines, dried fish skins were used in Britain and Europe as a weak source of light. This experimental form of illumination avoided the necessity of using candles which risked sparking explosions of firedamp. Another safe source of illumination in mines was bottles containing fireflies. In 1920, the American zoologist E. Newton Harvey published a monograph, "The Nature of Animal Light", summarizing early work on bioluminescence. Harvey notes that Aristotle mentions light produced by dead fish and flesh, and that both Aristotle and Pliny the Elder (in his "Natural History") mention light from damp wood. He also records that Robert Boyle experimented on these light sources, and showed that both they and the glowworm require air for light to be produced. Harvey notes that in 1753, J. Baker identified the flagellate "Noctiluca" "as a luminous animal" "just visible to the naked eye", and in 1854 Johann Florian Heller (1813–1871) identified strands (hyphae) of fungi as the source of light in dead wood.


title: Bioluminescence

text: Tuckey, in his posthumous 1818 "Narrative of the Expedition to the Zaire", described catching the animals responsible for luminescence. He mentions pellucids, crustaceans (to which he ascribes the milky whiteness of the water), and cancers (shrimps and crabs). Under the microscope he described the "luminous property" to be in the brain, resembling "a most brilliant amethyst about the size of a large pin's head".


title: Bioluminescence

text: While sailing in these latitudes on one very dark night, the sea presented a wonderful and most beautiful spectacle. There was a fresh breeze, and every part of the surface, which during the day is seen as foam, now glowed with a pale light. The vessel drove before her bows two billows of liquid phosphorus, and in her wake she was followed by a milky train. As far as the eye reached, the crest of every wave was bright, and the sky above the horizon, from the reflected glare of these livid flames, was not so utterly obscure, as over the rest of the heavens.


title: Bioluminescence

text: Darwin also observed a luminous "jelly-fish of the genus Dianaea", noting that: "When the waves scintillate with bright green sparks, I believe it is generally owing to minute crustacea. But there can be no doubt that very many other pelagic animals, when alive, are phosphorescent." He guessed that "a disturbed electrical condition of the atmosphere" was probably responsible. Daniel Pauly comments that Darwin "was lucky with most of his guesses, but not here", noting that biochemistry was too little known, and that the complex evolution of the marine animals involved "would have been too much for comfort".


title: Bioluminescence

text: Bioluminescence attracted the attention of the United States Navy in the Cold War, since submarines in some waters can create a bright enough wake to be detected; a German submarine was sunk in the First World War, having been detected in this way. The navy was interested in predicting when such detection would be possible, and hence guiding their own submarines to avoid detection.


title: Bioluminescence

text: Among the anecdotes of navigation by bioluminescence is one recounted by the Apollo 13 astronaut Jim Lovell, who as a navy pilot had found his way back to his aircraft carrier USS "Shangri-La" when his navigation systems failed. Turning off his cabin lights, he saw the glowing wake of the ship, and was able to fly to it and land safely.


title: Bioluminescence

text: The French pharmacologist Raphaël Dubois carried out work on bioluminescence in the late nineteenth century. He studied click beetles ("Pyrophorus") and the marine bivalve mollusc "Pholas dactylus". He refuted the old idea that bioluminescence came from phosphorus, and demonstrated that the process was related to the oxidation of a specific compound, which he named luciferin, by an enzyme. He sent Harvey siphons from the mollusc preserved in sugar. Harvey had become interested in bioluminescence as a result of visiting the South Pacific and Japan and observing phosphorescent organisms there. He studied the phenomenon for many years. His research aimed to demonstrate that luciferin, and the enzymes that act on it to produce light, were interchangeable between species, showing that all bioluminescent organisms had a common ancestor. However, he found this hypothesis to be false, with different organisms having major differences in the composition of their light-producing proteins. He spent the next 30 years purifying and studying the components, but it fell to the young Japanese chemist Osamu Shimomura to be the first to obtain crystalline luciferin. He used the sea firefly "Vargula hilgendorfii", but it was another ten years before he discovered the chemical's structure and published his 1957 paper "Crystalline Cypridina Luciferin". Shimomura, Martin Chalfie, and Roger Y. Tsien won the 2008 Nobel Prize in Chemistry for their 1961 discovery and development of green fluorescent protein as a tool for biological research.


title: Bioluminescence

text: Harvey wrote a detailed historical account on all forms of luminescence in 1957. An updated book on bioluminescence covering also the twentieth and early twenty-first century was published recently.


title: Bioluminescence

text: In 1932 E. N. Harvey was among the first to propose how bioluminescence could have evolved. In this early paper, he suggested that proto-bioluminescence could have arisen from respiratory chain proteins that hold fluorescent groups. This hypothesis has since been disproven, but it did lead to considerable interest in the origins of the phenomenon. Today, the two prevailing hypotheses (both concerning marine bioluminescence) are those put forth by Howard Seliger in 1993 and Rees et al. in 1998.


title: Bioluminescence

text: Seliger's theory identifies luciferase enzymes as the catalyst for the evolution of bioluminescent systems. It suggests that the original purpose of luciferases was as mixed-function oxygenases. As the early ancestors of many species moved into deeper and darker waters natural selection favored the development of increased eye sensitivity and enhanced visual signals. If selection were to favor a mutation in the oxygenase enzyme required for the breakdown of pigment molecules (molecules often associated with spots used to attract a mate or distract a predator) it could have eventually resulted in external luminescence in tissues.


title: Bioluminescence

text: Rees et al. use evidence gathered from the marine luciferin coelenterazine to suggest that selection acting on luciferins may have arisen from pressures to protect oceanic organisms from potentially deleterious reactive oxygen species (e.g. HO and O ). The functional shift from antioxidation to bioluminescence probably occurred when the strength of selection for antioxidation defense decreased as early species moved further down the water column. At greater depths exposure to ROS is significantly lower, as is the endogenous production of ROS through metabolism.


title: Bioluminescence

text: While popular at first, Seliger's theory has been challenged, particularly on the biochemical and genetic evidence that Rees examines. What remains clear, however, is that bioluminescence has evolved independently at least 40 times. Bioluminescence in fish began at least by the Cretaceous period. About 1,500 fish species are known to be bioluminescent; the capability evolved independently at least 27 times. Of these, 17 involved the taking up of bioluminous bacteria from the surrounding water while in the others, the intrinsic light evolved through chemical synthesis. These fish have become surprisingly diverse in the deep ocean and control their light with the help of their nervous system, using it not just to lure prey or hide from predators, but also for communication.


title: Bioluminescence

text: All bioluminescent organisms have in common that the reaction of a "luciferin" and oxygen is catalyzed by a luciferase to produce light. McElroy and Seliger proposed in 1962 that the bioluminescent reaction evolved to detoxify oxygen, in parallel with photosynthesis.


title: Bioluminescence

text: Thuesen, Davis et al. showed in 2016 that bioluminescence has evolved independently 27 times within 14 fish clades across ray-finned fishes.


title: Bioluminescence

text: Bioluminescence is a form of chemiluminescence where light energy is released by a chemical reaction. This reaction involves a light-emitting pigment, the luciferin, and a luciferase, the enzyme component. Because of the diversity of luciferin/luciferase combinations, there are very few commonalities in the chemical mechanism. From currently studied systems, the only unifying mechanism is the role of molecular oxygen; often there is a concurrent release of carbon dioxide (CO). For example, the firefly luciferin/luciferase reaction requires magnesium and ATP and produces CO, adenosine monophosphate (AMP) and pyrophosphate (PP) as waste products. Other cofactors may be required, such as calcium (Ca) for the photoprotein aequorin, or magnesium (Mg) ions and ATP for the firefly luciferase. Generically, this reaction can be described as:


title: Bioluminescence

text: Instead of a luciferase, the jellyfish "Aequorea victoria" makes use of another type of protein called a photoprotein, in this case specifically aequorin. When calcium ions are added, rapid catalysis creates a brief flash quite unlike the prolonged glow produced by luciferase. In a second, much slower step, luciferin is regenerated from the oxidized (oxyluciferin) form, allowing it to recombine with aequorin, in preparation for a subsequent flash. Photoproteins are thus enzymes, but with unusual reaction kinetics. Furthermore, some of the blue light released by aequorin in contact with calcium ions is absorbed by a green fluorescent protein, which in turn releases green light in a process called resonant energy transfer.


title: Bioluminescence

text: Overall, bioluminescence has arisen over 40 times in evolutionary history. In evolution, luciferins tend to vary little: one in particular, coelenterazine, is the light emitting pigment for nine phyla (groups of very different organisms), including polycystine radiolaria, Cercozoa (Phaeodaria), protozoa, comb jellies, cnidaria including jellyfish and corals, crustaceans, molluscs, arrow worms and vertebrates (ray-finned fish). Not all these organisms synthesise coelenterazine: some of them obtain it through their diet. Conversely, luciferase enzymes vary widely and tend to be different in each species.


title: Bioluminescence

text: Bioluminescence occurs widely among animals, especially in the open sea, including fish, jellyfish, comb jellies, crustaceans, and cephalopod molluscs; in some fungi and bacteria; and in various terrestrial invertebrates including insects. In marine coastal habitats, about 2.5% of organisms are estimated to be bioluminescent, whereas in pelagic habitats in the eastern Pacific, about 76% of the main taxa of deep-sea animals have been found to be capable of producing light. Most marine light-emission is in the blue and green light spectrum. However, some loose-jawed fish emit red and infrared light, and the genus "Tomopteris" emits yellow light.


title: Bioluminescence

text: The most frequently encountered bioluminescent organisms may be the dinoflagellates in the surface layers of the sea, which are responsible for the sparkling phosphorescence sometimes seen at night in disturbed water. At least 18 genera exhibit luminosity. A different effect is the thousands of square miles of the ocean which shine with the light produced by bioluminescent bacteria, known as mareel or the milky seas effect.


title: Bioluminescence

text: Bioluminescence is abundant in the pelagic zone, with the most concentration at depths devoid of light and surface waters at night. These organisms participate in diurnal vertical migration from the dark depths to the surface at night, dispersing the population of bioluminescent organisms across the pelagic water column. The dispersal of bioluminescence across different depths in the pelagic zone has been attributed to the selection pressures imposed by predation and the lack of places to hide in the open sea. In depths where sunlight never penetrates, often below 200m, the significance of bioluminescent is evident in the retainment of functional eyes for organisms to detect bioluminescence.


title: Bioluminescence

text: Organisms often produce bioluminescence themselves, rarely do they generate it from outside phenomena. However, there are occasions where bioluminescence is produced by bacterial symbionts that have a symbiotic relationship with the host organism. Although many luminous bacteria in the marine environment are free-living, a majority are found in symbiotic relationships that involve fish, squids, crustaceans etc. as hosts. Most luminous bacterial inhabit the marine sea, with "Photobacterium" and "Vibrio" genera dominating the marine environment.


title: Bioluminescence

text: In the symbiotic relationship, bacterium benefit from having a source of nourishment and a refuge to grow. Hosts obtain these bacterial symbionts either from the environment, spawning, or the luminous bacterium is evolving with their host. Coevolutionary interactions are suggested as host organisms’ anatomical adaptations have become specific to only certain luminous bacteria, to suffice ecological dependence of bioluminescence.


title: Bioluminescence

text: Bioluminescence is widely studied amongst species located in the mesopelagic zone, but the benthic zone at mesopelagic depths has remained widely unknown. Benthic habitats at depths beyond the mesopelagic are also poorly understood due to the same constraints. Unlike the pelagic zone where the emission of light is undisturbed in the open sea, the occurrence of bioluminescence in the benthic zone is less common. It has been attributed to the blockage of emitted light by a number of sources such as the sea floor, and inorganic and organic structures. Visual signals and communication that is prevalent in the pelagic zone such as counterillumination may not be functional or relevant in the benthic realm. Bioluminescence in bathyal benthic species still remains poorly studied due to difficulties of the collection of species at these depths.


title: Bioluminescence

text: Bioluminescence has several functions in different taxa. Steven Haddock et al. (2010) list as more or less definite functions in marine organisms the following: defensive functions of startle, counterillumination (camouflage), misdirection (smoke screen), distractive body parts, burglar alarm (making predators easier for higher predators to see), and warning to deter settlers; offensive functions of lure, stun or confuse prey, illuminate prey, and mate attraction/recognition. It is much easier for researchers to detect that a species is able to produce light than to analyze the chemical mechanisms or to prove what function the light serves. In some cases the function is unknown, as with species in three families of earthworm (Oligochaeta), such as "Diplocardia longa" where the coelomic fluid produces light when the animal moves. The following functions are reasonably well established in the named organisms.


title: Bioluminescence

text: In many animals of the deep sea, including several squid species, bacterial bioluminescence is used for camouflage by counterillumination, in which the animal matches the overhead environmental light as seen from below. In these animals, photoreceptors control the illumination to match the brightness of the background. These light organs are usually separate from the tissue containing the bioluminescent bacteria. However, in one species, "Euprymna scolopes", the bacteria are an integral component of the animal's light organ.


title: Bioluminescence

text: Bioluminescence is used in a variety of ways and for different purposes. The cirrate octopod "Stauroteuthis syrtensis" uses emits bioluminescence from its sucker like structures. These structures are believed to have evolved from what are more commonly known as octopus suckers. They do not have the same function as the normal suckers because they no longer have any handling or grappling ability due its evolution of photophores. The placement of the photophores are within the animals oral reach, which leads researchers to suggest that it uses it bioluminescence to capture and lure prey.


title: Bioluminescence

text: Fireflies use light to attract mates. Two systems are involved according to species; in one, females emit light from their abdomens to attract males; in the other, flying males emit signals to which the sometimes sedentary females respond. Click beetles emit an orange light from the abdomen when flying and a green light from the thorax when they are disturbed or moving about on the ground. The former is probably a sexual attractant but the latter may be defensive. Larvae of the click beetle "Pyrophorus nyctophanus" live in the surface layers of termite mounds in Brazil. They light up the mounds by emitting a bright greenish glow which attracts the flying insects on which they feed.


title: Bioluminescence

text: In the marine environment, use of luminescence for mate attraction is chiefly known among ostracods, small shrimplike crustaceans, especially in the family Cyprididae. Pheromones may be used for long-distance communication, with bioluminescence used at close range to enable mates to "home in". A polychaete worm, the Bermuda fireworm creates a brief display, a few nights after the full moon, when the female lights up to attract males.


title: Bioluminescence

text: The defense mechanisms for bioluminescent organisms can come in multiple forms; startling prey, counterillumination, smoke screen or misdirection, distractive body parts, burglar alarm, sacrificial tag or warning coloration. The shrimp family Oplophoridae Dana use their bioluminescence as a way of startling the predator that is after them. "Acanthephyra purpurea", within the Oplophoridae family, uses its photophores to emit light, and can secrete a bioluminescent substance when in the presence of a predator. This secretory mechanism is common among prey fish.


title: Bioluminescence

text: Many cephalopods, including at least 70 genera of squid, are bioluminescent. Some squid and small crustaceans use bioluminescent chemical mixtures or bacterial slurries in the same way as many squid use ink. A cloud of luminescent material is expelled, distracting or repelling a potential predator, while the animal escapes to safety. The deep sea squid "Octopoteuthis deletron" may autotomise portions of its arms which are luminous and continue to twitch and flash, thus distracting a predator while the animal flees.


title: Bioluminescence

text: Dinoflagellates may use bioluminescence for defense against predators. They shine when they detect a predator, possibly making the predator itself more vulnerable by attracting the attention of predators from higher trophic levels. Grazing copepods release any phytoplankton cells that flash, unharmed; if they were eaten they would make the copepods glow, attracting predators, so the phytoplankton's bioluminescence is defensive. The problem of shining stomach contents is solved (and the explanation corroborated) in predatory deep-sea fishes: their stomachs have a black lining able to keep the light from any bioluminescent fish prey which they have swallowed from attracting larger predators.


title: Bioluminescence

text: The sea-firefly is a small crustacean living in sediment. At rest it emits a dull glow but when disturbed it darts away leaving a cloud of shimmering blue light to confuse the predator. During World War II it was gathered and dried for use by the Japanese army as a source of light during clandestine operations.


title: Bioluminescence

text: The larvae of railroad worms ("Phrixothrix") have paired photic organs on each body segment, able to glow with green light; these are thought to have a defensive purpose. They also have organs on the head which produce red light; they are the only terrestrial organisms to emit light of this color.


title: Bioluminescence

text: Aposematism is a widely used function of bioluminescence, providing a warning that the creature concerned is unpalatable. It is suggested that many firefly larvae glow to repel predators; some millipedes glow for the same purpose. Some marine organisms are believed to emit light for a similar reason. These include scale worms, jellyfish and brittle stars but further research is needed to fully establish the function of the luminescence. Such a mechanism would be of particular advantage to soft-bodied cnidarians if they were able to deter predation in this way. The limpet "Latia neritoides" is the only known freshwater gastropod that emits light. It produces greenish luminescent mucus which may have an anti-predator function. The marine snail "Hinea brasiliana" uses flashes of light, probably to deter predators. The blue-green light is emitted through the translucent shell, which functions as an efficient diffuser of light.


title: Bioluminescence

text: Communication in the form of quorum sensing plays a role in the regulation of luminescence in many species of bacteria. Small extracellularly secreted molecules stimulate the bacteria to turn on genes for light production when cell density, measured by concentration of the secreted molecules, is high.


title: Bioluminescence

text: Pyrosomes are colonial tunicates and each zooid has a pair of luminescent organs on either side of the inlet siphon. When stimulated by light, these turn on and off, causing rhythmic flashing. No neural pathway runs between the zooids, but each responds to the light produced by other individuals, and even to light from other nearby colonies. Communication by light emission between the zooids enables coordination of colony effort, for example in swimming where each zooid provides part of the propulsive force.


title: Bioluminescence

text: Some bioluminous bacteria infect nematodes that parasitize Lepidoptera larvae. When these caterpillars die, their luminosity may attract predators to the dead insect thus assisting in the dispersal of both bacteria and nematodes. A similar reason may account for the many species of fungi that emit light. Species in the genera "Armillaria", "Mycena", "Omphalotus", "Panellus", "Pleurotus" and others do this, emitting usually greenish light from the mycelium, cap and gills. This may attract night-flying insects and aid in spore dispersal, but other functions may also be involved.


title: Bioluminescence

text: "Quantula striata" is the only known bioluminescent terrestrial mollusc. Pulses of light are emitted from a gland near the front of the foot and may have a communicative function, although the adaptive significance is not fully understood.


title: Bioluminescence

text: Bioluminescence is used by a variety of animals to mimic other species. Many species of deep sea fish such as the anglerfish and dragonfish make use of aggressive mimicry to attract prey. They have an appendage on their heads called an esca that contains bioluminescent bacteria able to produce a long-lasting glow which the fish can control. The glowing esca is dangled or waved about to lure small animals to within striking distance of the fish.


title: Bioluminescence

text: The cookiecutter shark uses bioluminescence to camouflage its underside by counterillumination, but a small patch near its pectoral fins remains dark, appearing as a small fish to large predatory fish like tuna and mackerel swimming beneath it. When such fish approach the lure, they are bitten by the shark.


title: Bioluminescence

text: Female "Photuris" fireflies sometimes mimic the light pattern of another firefly, "Photinus", to attract its males as prey. In this way they obtain both food and the defensive chemicals named lucibufagins, which "Photuris" cannot synthesize.


title: Bioluminescence

text: South American giant cockroaches of the genus "Lucihormetica" were believed to be the first known example of defensive mimicry, emitting light in imitation of bioluminescent, poisonous click beetles. However, doubt has been cast on this assertion, and there is no conclusive evidence that the cockroaches are bioluminescent.


title: Bioluminescence

text: While most marine bioluminescence is green to blue, some deep sea barbeled dragonfishes in the genera "Aristostomias", "Pachystomias" and "Malacosteus" emit a red glow. This adaptation allows the fish to see red-pigmented prey, which are normally invisible to other organisms in the deep ocean environment where red light has been filtered out by the water column. The fish is able to utilize the longer wavelength to act as a spotlight for its prey that only it is able to see. In addition to the utilization of the light for predation, it has been hypothesized that the fish use this to communicate with each other to find potential mates. The ability of the fish to see this light is explained by the presence of specialized rhodopsin pigment. The angler siphonophore (Erenna), also utilizes red bioluminescence in appendages to lure fish.


title: Bioluminescence

text: The mechanism of light creation is through a suborbital photophore that utilizes gland cells which produce exergonic chemical reactions that produce light with a longer, red wavelength. The dragonfish species which produce the red light also produce blue light in photophore on the dorsal area. The main function of this is to alert the fish to the presence of its prey. The additional pigment is thought to be assimilated from chlorophyll derivatives found in the copepods which form part of its diet.


title: Bioluminescence

text: Bioluminescent organisms are a target for many areas of research. Luciferase systems are widely used in genetic engineering as reporter genes, each producing a different color by fluorescence, and for biomedical research using bioluminescence imaging. For example, the firefly luciferase gene was used as early as 1986 for research using transgenic tobacco plants. "Vibrio" bacteria symbiose with marine invertebrates such as the Hawaiian bobtail squid ("Euprymna scolopes"), are key experimental models for bioluminescence. Bioluminescent activated destruction is an experimental cancer treatment.


title: Bioluminescence

text: "In Vivo" luminescence cell and animal imaging uses dyes and fluorescent proteins as chromophores. The characteristics of each chromophore determine which cell area(s) will be targeted and illuminated.


title: Bioluminescence

text: The structures of photophores, the light producing organs in bioluminescent organisms, are being investigated by industrial designers. Engineered bioluminescence could perhaps one day be used to reduce the need for street lighting, or for decorative purposes if it becomes possible to produce light that is both bright enough and can be sustained for long periods at a workable price. The gene that makes the tails of fireflies glow has been added to mustard plants. The plants glow faintly for an hour when touched, but a sensitive camera is needed to see the glow. University of Wisconsin–Madison is researching the use of genetically engineered bioluminescent E. coli bacteria, for use as bioluminescent bacteria in a light bulb.


title: Bioluminescence

text: An iGEM team from Cambridge (England) has started to address the problem that luciferin is consumed in the light-producing reaction by developing a genetic biotechnology part that codes for a luciferin regenerating enzyme from the North American firefly.


title: Bioluminescence

text: In 2016, Glowee, a French company started selling bioluminescent lights for shop fronts and street signs, for use between 1 and 7 in the morning when the law forbids use of electricity for this purpose. They used the bioluminescent bacterium "Aliivibrio fischeri", but the maximum lifetime of their product was three days.


title: Bioluminescence

text: In April 2020, plants were genetically engineered to glow more brightly using genes from the bioluminescent mushroom "Neonothopanus nambi" to convert caffeic acid into luciferin.


title: Bioluminescence

text: ATP bioluminescence is the process in which ATP is used to generate luminescence in an organism. It proves to be a very good biosensor to test cell viability. Optical biosensors include process of measurement of "luminescence, fluorescence absorbance or emission". Through these measurements, quantitative measurement of ATP bioluminescence is applied to detect existence of living microbes only. Since the method is quick and convenient, it results in real-time data. It is faster, economical and easier to work with. Optical biosensors sense the observed optical signal based on measuring the photons involved in the phenomenon ("spiking") It depends on the interaction of microbes with analytes. Thus, it is correlated with the concentration of the microbial population which is determined through this method.


title: Bioluminescence

text: In ATP bioluminescence, it is assumed that all living cells in the same have the same amount of ATP over time during the chemical reaction between luciferin, luciferase to produce ATP, This is done in order to measure the viability of the cell and allows the researcher to measure the amount of living and dead cells in the sample on basis of presence or absence of ATP. Living cells that contain ATP produce a bioluminescent flash due to the "luciferin-luciferase reaction" in presence of ATP. Dead cells do not produce any bioluminescence due to absence of ATP The amount of the intensity of the signal is constant for each living cell in a healthy sample. In this way, the overall number of living cells within a sample is determined.


title: Bioluminescence

text: ATP, which is a fundamental compound in the luciferase reaction, is utilized and in the second step, oxyluciferin is produced. The oxyluciferin is produced in an excited state, which produces light when it goes back to ground state. The light emitted is detected by a luminometer. Concentration of the ATP is directly proportional to the expressed light measured as Relative Light Units (RLU). A receiver operating characteristic (ROC) is used to calculate the sensitivity and specificity of the measurements. There is direct correlation between luminescence intensity and concentration of standard ATP. There is a direct correlation between bioluminescence and colony forming unit (CFU). Thus, concentration of standard ATP and CFU gives a standard correlation. In this way, ATP is measured and microbial population is determined through bioluminescence.


title: Bioluminescence

text: However, it is important to keep in mind that different types of microbial populations are determined through different sets of ATP assays using other substrates and reagents. "Renilla" and "Gaussia" based cell viability assays use the substrate coelenterazine.


title: Malaysian ringgit

text: The Malaysian ringgit (; plural: ringgit; symbol: RM; currency code: MYR; Malay name: "Ringgit Malaysia"; formerly the Malaysian dollar) is the currency of Malaysia. It is divided into 100 "sen" (formerly "cents"). The ringgit is issued by the Central Bank of Malaysia.


title: Malaysian ringgit

text: The word "ringgit" is an obsolete term for "jagged" in the Malay language. The word was originally used to refer to the serrated edges. The first European coins to circulate widely in the region were Spanish "pieces of eight" or "cob", their crude appearance resembling stones, hence the word jagged. The availability and circulation of this Spanish currency were due to the Spanish controlling nearby Philippines.


title: Malaysian ringgit

text: An early printed source, the "Dictionary of the Malayan Language" from 1812 had already referred to the ringgit as a unit of money.


title: Malaysian ringgit

text: In modern usage, "ringgit" is used almost solely for the currency. Due to the common heritage of the three modern currencies, the Singapore dollar and the Brunei dollar are also called "ringgit" in Malay (currencies such as the US and Australian dollars are translated as "dolar"), although nowadays the Singapore dollar is more commonly called "dolar" in Malay. To differentiate between the three currencies, the Malaysian currency is referred to as "Ringgit Malaysia", hence the official abbreviation and currency symbol "RM". Internationally, the ISO 4217 currency code for Malaysian ringgit is "MYR".


title: Malaysian ringgit

text: The Malay names "ringgit" and "sen" were officially adopted as the sole official names in August 1975. Previously they had been known officially as dollars and cents in English and ringgit and sen in Malay, and in some parts of the country this usage continues. In the northern states of Peninsular Malaysia, denominations of 10 "sen" are called "kupang" in Malay and called "pua̍t" (鏺/鈸) in Penang Hokkien which is thought to be derived from the Thai word "baht". e.g. 50 "sen" is "lima kupang" in Malay or 'samah' in Malay Kelantan dialect and "gōo-pua̍t" (五鏺/鈸) in Hokkien. The Tamil speaking communities in Malaysia use "veḷḷi" (வெள்ளி) meaning "silver" in Tamil to refer to "ringgit", while for "sen", the word "kācu" (காசு) is used, from which the English word "cash" is derived.


title: Malaysian ringgit

text: The Spanish-American silver dollar brought over by the Manila galleons was the primary currency for international trade, used in Asia and the Americas from the 16th to 19th centuries; it was eventually called the "ringgit". The various dollars introduced in the 19th century were itself derived from the Spanish dollar: the Straits dollar, Sarawak dollar and the British North Borneo dollar. From these dollars were derived their successor currencies the Malayan dollar and the Malaya and British Borneo dollar, and eventually the modern-day Malaysian ringgit, Singapore dollar and Brunei dollar.


title: Malaysian ringgit

text: On 12 June 1967, the Malaysian dollar, issued by the new central bank, Central Bank of Malaysia, replaced the Malaya and British Borneo dollar at par. The new currency retained all denominations of its predecessor except the $10,000 denomination, and also brought over the colour schemes of the old dollar. Over the course of the following decades, minor changes were made to the notes and coins issued, from the introduction of the M$1 coin in 1967, to the demonetization of RM500 and RM1,000 notes in 1999.


title: Malaysian ringgit

text: As the Malaysian dollar replaced the Malaya and British Borneo dollar at par and Malaysia was a participating member of the sterling area, the new dollar was originally valued at dollars per 1 British pound sterling; in turn, £1 = US$2.80 so that US$1 = M$3.06. In November 1967, five months after the introduction of the Malaysian dollar, the pound was devalued by 14.3% from US$2.80 to US$2.40, leading to a collapse in confidence for the sterling area and its demise in 1972. The new currency stayed pegged to the U.S. dollar at US$1 = M$3.06, but earlier notes of the Malaya and British Borneo dollar were devalued from US$2.80 to US$2.40 for 8.57 dollars; consequently these notes were reduced in value to 85 cents per dollar.


title: Malaysian ringgit

text: Despite the emergence of new currencies in Malaysia, Singapore and Brunei, the Interchangeability Agreement which the three countries adhered to as original members of the currency union meant the Malaysian dollar was exchangeable at par with the Singapore dollar and Brunei dollar. This ended on 8 May 1973, when the Malaysian government withdrew from the agreement. The Monetary Authority of Singapore and the Brunei Currency and Monetary Board still maintain the interchangeability of their two currencies, as of 2021.


title: Malaysian ringgit

text: In 1993, the currency symbol "RM" (Ringgit Malaysia) was introduced to replace the use of the dollar sign "$" (or "M$").


title: Malaysian ringgit

text: Between 1995 and 1997, the ringgit was trading as a free float currency at around 2.50 to the US dollar, but following the onset of the 1997 Asian financial crisis, the ringgit witnessed major dips to under 3.80 MYR/USD by the end of 1997 as a result of capital flight. During the first half of 1998, the currency fluctuated between 3.80 and 4.40 MYR/USD, before the Central Bank of Malaysia moved to peg the ringgit to the US dollar in September 1998, maintaining its 3.80 MYR/USD value while remaining floated against other currencies. In addition, the ringgit was designated non-tradeable outside of Malaysia in 1998 to stem the flow of money out of the country.


title: Malaysian ringgit

text: While the printing of RM500 and RM1,000 notes had ceased in 1996 in response to risks of money laundering and capital flight, the underestimated effects of the financial crisis prompted the central bank to completely discontinue the use of the notes by demonetising the remaining notes in circulation beginning 1 July 1999. The two denominations hereby ceased to be legal tender and were only exchangeable directly at the central bank; at the time of the demonetization, RM500 and RM1,000 notes were each worth approximately US$130 and US$260 respectably, based on the 3.80 MYR/USD peg rate. Despite these measures, some 7.6% of RM500 notes and 0.6% of RM1,000 notes remain in circulation as of 30 January 2011. During a 2011 parliamentary session, then Deputy Finance Minister Donald Lim Siang Chai asserted that a total of 150,599 and 26,018 pieces of RM500 and RM1,000 notes (RM75,299,500 worth of RM500 notes and RM26,018,000.00 worth of RM1,000 notes) have yet to be withdrawn through the central bank.


title: Malaysian ringgit

text: The ringgit lost 50% of its value against the US dollar between 1997 and 1998, and suffered general depreciation against other currencies between December 2001 and January 2005. As of 4 September 2008, the ringgit had yet to regain its value circa 2001 against the Singapore dollar (2.07 to 2.40 MYR/SGD), the euro (3.40 to 4.97 MYR/EUR), the Australian dollar (1.98 to 2.80 MYR/AUD), and the British pound (5.42 to 6.10 MYR/GBP).


title: Malaysian ringgit

text: On 21 July 2005, Central Bank of Malaysia announced the end of the peg to the US dollar immediately after China's announcement of the end of the renminbi peg to the US dollar. According to Bank Negara, Malaysia allows the ringgit to operate in a managed float against several major currencies. This has resulted in the value of the ringgit rising closer to its perceived market value, although Central Bank of Malaysia has intervened in financial markets to maintain stability in the trading level of the ringgit, a task made easier by the fact that the ringgit was pegged and has remained non-tradeable outside Malaysia since 1998.


title: Malaysian ringgit

text: Following the end of the currency peg, the ringgit appreciated to as high as 3.16 MYR/USD in April 2008. The ringgit had also enjoyed a period of appreciation against the Hong Kong dollar (from 0.49 to 0.44 MYR/HKD) and the renminbi (0.46 to 0.45 MYR/CNY) as recently as May 2008. The initial stability of the ringgit in the late-2000s had led to considerations to reintroduce the currency to foreign trading after over a decade of being non-internationalised. In a CNBC interview in September 2010, Najib Tun Razak, the then Prime Minister and Finance Minister of Malaysia, was quoted in stating that the government was planning the reentry of the ringgit into off-shore trading if the move will help the economy, with the condition that rules and regulations were put in place to prevent abuses. Despite considerations, the ringgit has continued to remain non-internationalised in a deliberate move to continue discouraging off-shore trading of the currency.


title: Malaysian ringgit

text: Political uncertainty following the country's 2008 general election and the 2008 Permatang Pauh by-election, falling crude oil prices in the late-2000s, and the lack of intervention by the Central Bank of Malaysia to increase already low interest rates (which remained at 3.5% between April 2006 and November 2008) led to a slight fall of the ringgit's value against the US dollar between May and July 2008, followed by a sharper drop between August and September of the same year. As a result, the US dollar appreciated significantly to close at 3.43 MYR/USD as of 4 September 2008, while other major currencies, including the renminbi and Hong Kong dollar, followed suit. The ringgit spiked at 3.73 MYR/USD by March 2009, before gradually recovering to 3.00 MYR/USD by mid-2011 and normalising at around 3.10 MYR/USD between 2011 and 2014.


title: Malaysian ringgit

text: The ringgit experienced more acute plunges in the value since mid-2014 following the escalation of the 1Malaysia Development Berhad scandal that raised allegations of political channeling of billions of ringgit to off-shore accounts, and uncertainty from the 2015–16 Chinese stock market turbulence and the effects of the 2016 United States presidential election results. The currency's value fell from an average of 3.20 MYR/USD in mid-2014 to around 3.70 MYR/USD by early 2015; with China being Malaysia's largest trading partner, a Chinese stock market crash in June 2015 triggered another plunge in value for the ringgit, which reached levels unseen since 1998 at lows of 4.43 MYR/USD in September 2015, before stabilising around 4.10 to 4.20 to the US dollar soon after; the currency later plummeted and hover below the 1998 lows at 4.40 and 4.50 MYR/USD, following the wake of the victory of pro-protectionist Donald Trump in the 2016 United States presidential election, which has raised questions of the United States' participation in the Trans-Pacific Partnership (TPP) (which Malaysia is a signatory of, and the United States had promptly pulled out from in January 2017) and Malaysia–United States trade as a whole (as the United States is among Malaysia's largest trading partners).


title: Malaysian ringgit

text: In response to the sharp drop of the ringgit in November 2016, Central Bank of Malaysia began a series of tougher crackdowns on under-the-counter non-deliverable forward trading of the ringgit in order to curb currency speculation. Since then, the currency has seen a steady but consistent rate of appreciation against the US dollar, with significant increases since early-November 2017 following reports of positive economic performance, the restructuring of the TPP into the Comprehensive and Progressive Agreement for Trans-Pacific Partnership and increasing global oil prices. After appreciating as high as 3.86 to the US dollar as of early April 2018, the value dropped to around 4.18 MYR/USD by the end of October 2018 following increasing trade war tensions in response to the China–United States trade war, selloff panic from other emerging markets, as well as uncertainty in economic policy following an upset by the Pakatan Harapan coalition in the 2018 general election. With the exception of the Euro, the currency's has also seen some recovery of value to pre-late 2016 levels against other major currencies, including the renminbi, British pound, Australian dollar, Japanese yen and Singaporean dollar, but remains less valuable overall than before the end of 2013.


title: Malaysian ringgit

text: The first series of sen coins were introduced in 1967 in denominations of 1, 5, 10, 20 and 50 sen, followed by the introduction of the 1 ringgit coin (which used the $ symbol and is the largest coin in the series) in 1971. While varied by diameters, virtually all the coins were minted in near-consistent obverse and reverse designs and were very generic, with the obverse depicting the then recently completed Malaysian Houses of Parliament and the federal star and crescent moon from the canton of the Malaysian flag. All coins were minted from cupronickel, the only exception being the 1 sen coin, which was first composed from bronze between 1967 and 1972, then in steel clad with copper from 1973 onwards. The 50 sen coin is the only one in the series to undergo a redesign, a minor 1971 modification on its edge to include ""Bank Negara Malaysia"" letterings. All coins have the initials GC on the reverse, below the Parliament House. It stands for Geoffrey Colley, Malaysia first coin series' designer. The 1 ringgit coin was never popular at the time due to being in conflict with a banknote of equal face value, similar to the current situation regarding the 1 dollar coin of the United States dollar.


title: Malaysian ringgit

text: The coins of this first series were identical in size and composition to those of the former Malaya and British Borneo dollar. Though the Malayan currency union coins were withdrawn, they still appear in circulation on very rare occasions.


title: Malaysian ringgit

text: Minting of the first sen series ended in 1989, when the second series was introduced. The older coins remain legal tender as of 2019, but have steadily declined in number and are seldom seen in circulation in Malaysia.


title: Malaysian ringgit

text: The second series of sen coins entered circulation in late 1989, sporting completely redesigned observes and reverses, but predominantly retaining the design of edges, diameters and composition of the previous series' coins previous to 1989, the 1 ringgit coin being the exception. Changes include the depiction of items of Malay culture on the obverse, such as a local mancala game board called congkak on the 10 sen and the wau bulan or "moon kite" on the 50 sen among other things, as well as the inclusion of a Hibiscus rosa-sinensis (Malay: "Bunga Raya"), the national flower of Malaysia, on the upper half of the reverse. The second series was designed by Low Yee Kheng.


title: Malaysian ringgit

text: In addition to changes on its observe and reverse, the size of the 1 ringgit coin was also reduced from a diameter of 33 mm to 24 mm, and was minted from an alloy of copper, zinc and tin, as opposed to the first series' cupronickel. The $ symbol was brought over to the new coin, but was dropped in favour of "RINGGIT" for coins minted from 1993 onwards. On 7 December 2005, the 1 ringgit coin was demonetised and withdrawn from circulation. This was partly due to problems with standardisation (two different versions of the second series coin were minted) and forgery.


title: Malaysian ringgit

text: As of 1 April 2008, a rounding mechanism of prices to the nearest 5 sen, applied to the total bill only, is in force, which was first announced in 2007 by Central Bank of Malaysia, in an attempt to render the 1 sen coin irrelevant. Individual items and services can still be priced in multiples of 1 sen with the final totalled rounded to the nearest 5 sen. For example, purchasing two items priced RM4.88 and RM3.14, totalling RM8.02, would then be rounded to RM8.00. If each item had been individually rounded (to RM4.90 and RM3.15 respectively) the incorrect total would have been RM8.05. In practice, individual items will probably remain priced at so-called "price points" (or psychological pricing and odd-number pricing) ending in 98 and 99 to maximise rounding gains for the vendor, especially in the case of single item purchases. Existing 1 sen coins in circulation remain legal tender for payments up to RM2.00.


title: Malaysian ringgit

text: The third series of coins were announced on 25 July 2011, first being issued as commemorative coins to mark their release on 16 January 2012. The third series carry a theme named "Distinctively Malaysia" and are inspired from motifs of flora and fauna drawn from various cultures in Malaysia to "reflect the diversity and richness of Malaysia's national identity". The denominations issued are 5, 10, 20 and 50 sen. On 24 October 2011, Deputy Finance Minister Datuk Donald Lim named Poogsan Corporation of South Korea as the series' coin suppliers and the coins are minted at the Bank Negara Mint in Shah Alam.


title: Malaysian ringgit

text: According to Lim, costs in producing the coins will be reduced by 49% due to the change in metal composition. Other changes in the series include the diameter, the colour on the 20- and 50 sen coins (from silver to yellow) and a redesign on the obverse (featuring different motifs for each denomination), fourteen dots symbolising the thirteen states and the collective Federal Territories, and five horizontal lines indicating the five principles of Rukunegara.


title: Malaysian ringgit

text: The 50-cent coin is more distinctive than the other denominations. The round shape of the coin has nine indentations, forgoing the original "BANK NEGARA MALAYSIA" lettering. The obverse does not feature the five horizontal lines, but instead a latent image security feature is placed over the coin, where lettering of the denomination "50" and "SEN" can be seen when the coin is tilted slightly.


title: Malaysian ringgit

text: The 20 sen and 50 sen coins look similar to €0.10 and €0.20 coin in size, edge design and colour, however they are only worth at €0.047 and €0.12 respectively.


title: Malaysian ringgit

text: Three denominations of gold bullion coins, the "Kijang Emas" (the kijang, a species of deer, being part of Central Bank of Malaysia's logo) are also issued, at the face value of RM 50, RM 100 and RM 200, weighing oz, oz and 1 oz (Troy ounce), respectively. It is minted by the Kilang Wang Central Bank of Malaysia and was launched on 17 July 2001 by Central Bank of Malaysia, making Malaysia the twelfth country to issue its own gold bullion coins. Like other bullion coins issued around the world, the Kijang Emas is primarily used as an investment rather than in day-to-day circulation. The purchase and resale price of Kijang Emas is determined by the prevailing international gold market price. Current price of the Kijang Emas is rated at RM 8266 for 1oz, RM 4211 for oz and RM 2144 for oz (November 17, 2020).


title: Malaysian ringgit

text: Central Bank of Malaysia first issued Malaysian dollar banknotes on 6 June 1967 in $1, $5, $10, $50 and $100 denominations. The $1000 denomination was first issued on 2 September 1968. The first Malaysian banknotes carried the image of Tuanku Abdul Rahman, the first Yang di-Pertuan Agong of Malaysia and bore the signature of Tun Ismail bin Mohamed Ali, the first Malaysian Governor of Central Bank of Malaysia. On 16 August 1972, Central Bank of Malaysia adopted official new spelling system of the national language, Bahasa Malaysia, into the printing of its currency notes while retaining the designs. The banknotes with new spellings are circulated alongside the old banknotes.


title: Malaysian ringgit

text: The second series was issued with Malaysian traditional ornamental designs, with the introduction of two new denomination, the $20 and $500 in 1982; later followed by $10, $50 and $100 in 1983, finally the $1, $5 & $1000 completed this series in 1984. Until 2010 the second series notes was still occasionally encountered.


title: Malaysian ringgit

text: Due to its unpopularity, the $20 (RM20) denominations were discontinued and gradually removed from circulation in 1995.


title: Malaysian ringgit

text: In 1999 the RM500 and RM1000 notes were discontinued and ceased to be legal tender. This was due because of the Asian monetary crisis of 1997 when huge amounts of ringgit were taken out of the country to be traded in these notes. In effect the notes were withdrawn out of circulation and the amount of ringgit taken out of the country in banknotes was limited to RM1000.


title: Malaysian ringgit

text: The third series was issued with designs in the spirit of Wawasan 2020 in 1996 in denominations of RM1, RM2, RM5, RM10, RM50 and RM100. The larger denomination RM50 and RM100 notes had an additional hologram strip to deter counterfeiters.


title: Malaysian ringgit

text: In 2004, Central Bank of Malaysia issued a new RM10 note with additional security features including the holographic strip previously only seen on the RM50 and RM100 notes. A new RM5 polymer banknote with a distinctive transparent window was also issued. Both new banknotes are almost identical to their original third series designs. At one time, Central Bank of Malaysia announced its intention to eventually phase out all paper notes and replace them with polymer notes.


title: Malaysian ringgit

text: In early 2008, the Bank released a newly designed RM50 banknote, which according to the Bank, were to enter general circulation beginning 30 January 2008. Earlier, 20,000 more such notes with special packaging were distributed by the bank on 26 December 2007.


title: Malaysian ringgit

text: The newly designed RM50 banknote retains the predominant colour of green-blue, but is designed in a new theme, dubbed the "National Mission", expressing the notion of Malaysia "[moving] the economy up the value chain", in accordance to Malaysia's economic transformation to higher value-added activities in agriculture, manufacturing and services sectors of the economy. The dominant intaglio portrait of the first Yang di-Pertuan Agong, Tuanku Abdul Rahman, is retained on the right and the national flower, the hibiscus, is presented in the center on the obverse of the note. Design patterns from songket weaving, which are in the background and edges of the banknote, are featured to reflect the traditional Malay textile handicraft and embroidery.


title: Malaysian ringgit

text: The first 50 million pieces of the new RM50 banknote features Malaysia's first Prime Minister, Tunku Abdul Rahman, at the historic declaration of Malaya's independence, and the logo of the 50th Anniversary of Independence on the reverse. Security features on the banknote include a watermarked portrait of the Yang di-Pertuan Agong, a security thread, micro letterings, fluorescent elements visible only under ultraviolet light, a multi coloured latent image which changes colour when viewed at different angles, and a holographic stripe at the side of the note and an image that is visible only via a moiré effect to prevent counterfeiting using photocopiers. Circulation for the first edition of this new RM50 banknote was eventually curtailed by the Central Bank due to the various Malaysia banks' automatic teller machines inability to accept it. The bank began to re-release the new series for general circulation beginning 15 July 2009 without the 50th Anniversary logo. This edition include new enhanced security features such as two color number fluorescents and security fibres.


title: Malaysian ringgit

text: In May 2011, Central Bank of Malaysia had announced that they will introduce a new series of banknotes to replace the current design that has been in circulation for around 15 years. The most highlighted part of the announcement is the re-introduction of the RM20 note, which was not included in the third series. The design of the new notes was announced on 21 December 2011, and the notes are expected to be put into circulation in the second half of 2012. The new series banknotes are legal tender and will co-circulate with the existing series. The existing series will be gradually phased out. All 4 series of banknotes (except 500, and 1000) are technically still legal tender, but some vendors may not accept the first and second series banknotes (rarely seen now). All banknote denominations in the new series will retain the portrait of the first Yang di-Pertuan Agong, Tuanku Abdul Rahman. The banknotes are supplied by Crane AB of Sweden, Giesecke & Devrient GmbH of Germany, Oberthur Technologies of France and Orell Fussli of Switzerland. They were put into circulation on 16 July 2012.


title: Malaysian ringgit

text: To commemorate the 1998 Commonwealth Games in Kuala Lumpur, a commemorative RM50 polymer banknote was issued on 1 June 1998. This was the first polymer banknote to be issued by Central Bank of Malaysia and was printed by Note Printing Australia (NPA). A total of 500,000 sets were issued. They were sold in special packaging and at a premium price of 80 ringgit. This note is hardly ever seen in normal usage, its use being a collector's commemorative.


title: Malaysian ringgit

text: On 21 December 2007, Central Bank of Malaysia issued a commemorative 50 ringgit banknote to commemorate Malaysia's 50th Anniversary of Independence. The design was that of the 50 ringgit banknote of the fourth series, except with the additional logo of the 50th Anniversary of Independence at the top right of the reverse side, and the inscription "1957-2007" also on the reverse side of the banknote. The regular 50 ringgit notes which were issued later from 2009 onward did not carry these additional design features


title: Malaysian ringgit

text: A total of 50 million banknotes which bore the commemorative design, with serial number letter prefixes from AA to AE, were issued. Subsequent regular banknotes had serial number prefixes from AF onward. Of the 50 million commemorative banknotes, the first 20,000 were sold with a special packaging at a premium price of 60 ringgit.


title: Malaysian ringgit

text: On 14 December, Central Bank of Malaysia announced the issue of two paper-polymer hybrid commemorative banknotes in conjunction with the sixtieth anniversary of the Signing of the Federation of Malaya Independence Agreement. The banknotes were in the denominations of 60 ringgit and 600 ringgit. The 60 ringgit note was also made available in a 3-in-1 format.


title: Malaysian ringgit

text: The 600 ringgit note is the largest legal tender banknote in terms of size to be issued in the world, measuring 370mm by 220mm.


title: Malaysian ringgit

text: The notes were released for sale online on 29 December 2017 at a premium, with the 60 ringgit note sold at 120 ringgit, the 3-in-1 60 ringgit note at 500 ringgit and the 600 ringgit note at 1,700 ringgit. The print run for the 60 ringgit note was 60,000 while that for both the 3-in-1 60 ringgit and 600 ringgit note were at 6,000.


title: Steele dossier

text: The Steele dossier, also known as the Trump–Russia dossier, is a controversial political opposition research report written from June to December 2016, containing allegations of misconduct, conspiracy, and cooperation between Donald Trump's presidential campaign and the government of Russia prior to and during the 2016 election campaign. Five years later, it was described by mainstream media as "largely discredited", "deeply flawed", and "largely unverified". Some aspects of the dossier have been corroborated, namely that Vladimir Putin favored Trump over Hillary Clinton, and that many Trump campaign officials and associates had numerous secretive contacts with Russian officials and spies.


title: Steele dossier

text: The dossier, leaked by "BuzzFeed News" in January 2017, without its author's permission, is an unfinished compilation of raw intelligence based on information from initially anonymous sources known to the author, counterintelligence specialist Christopher Steele. Steele, a former head of the Russia Desk for British intelligence (MI6), was writing the report for the private investigative firm Fusion GPS, who were paid by Hillary Clinton's campaign and the Democratic National Committee (DNC). The dossier's 17 reports allege that Trump campaign members and Russian operatives had conspired to cooperate in Russia's election interference to benefit Trump. It also alleges that Russia sought to damage Hillary Clinton's candidacy. "BuzzFeed"s decision to publish the reports without verifying their allegations was criticized by journalists but their decision was also defended in a court statement explaining that "BuzzFeed"s action was for the public good as the public had a right to know so it could "exercise effective oversight of the government".


title: Steele dossier

text: In June 2016, Fusion GPS subcontracted Steele's firm to compile the dossier. DNC officials denied knowing their attorney had contracted with Fusion GPS, and Steele asserted he was not aware the Clinton campaign was the recipient of his research until months after he contracted with Fusion GPS. While compiling the dossier, Steele passed some of his findings to both British and American intelligence services.


title: Steele dossier

text: The intelligence community and most experts have treated the dossier with caution due to its unverified allegations, that Trump denounced as fake news. The U.S. intelligence community took the allegations seriously, and the Federal Bureau of Investigation (FBI) investigated every line of the dossier and identified and spoke with at least two of Steele's sources. The Mueller Report contained passing references to some of the dossier's allegations but little mention of its more sensational claims.


title: Steele dossier

text: Many allegations in the dossier have been dismissed by authorities or remain unverified. While the dossier played a central role in the seeking of FISA warrants on Carter Page, it did not play any role in the intelligence community's assessment about Russian actions in the 2016 election, and it was not the trigger for the opening of the Russia investigation into whether the Trump campaign was coordinating with the Russian government's interference in the 2016 presidential election. The dossier is a factor in several conspiracy theories promoted by Trump and his supporters in the media and Congress.


title: Steele dossier

text: The opposition research conducted by Fusion GPS on Donald Trump was in two distinct operations, each with a different client. The first research operation, from October 2015 to May 2016, was domestic research funded by "The Washington Free Beacon". The second operation, from April 2016 to December 2016, was funded by the DNC and the Clinton campaign. Only the second operation involved the foreign research used to create the dossier.<ref name="Lima_10/27/2017"></ref> From April 2016 into early May, the "Washington Free Beacon" and the Clinton Campaign/DNC were independently both clients of Fusion GPS.


title: Steele dossier

text: An "agreed chronology" of dossier-related events is published in Appendix C of the "Gubarev v Orbis" case of October 30, 2020.<ref name="Judiciary.uk_10/30/2020"></ref>


title: Steele dossier

text: In October 2015, before the official start of the 2016 Republican primary campaign, the founders of Fusion GPS were seeking political work and wrote an email to "a big conservative donor they knew who disliked Trump, [and] they were hired". He arranged for them to use "The Washington Free Beacon", an American conservative political journalism website, for their general opposition research on several Republican presidential candidates, including Trump.<ref name="Mayer_11/25/2019"></ref> It is primarily funded by Republican donor Paul Singer.<ref name="Vogel_Haberman_10/27/2017"></ref> The "Free Beacon" and Singer were "part of the conservative never-Trump movement". Although Singer was a big supporter of Marco Rubio, Rubio denied any involvement in Fusion GPS's initial research and hiring.


title: Steele dossier

text: Early in their investigation, they received help from investigative reporter Wayne Barrett, who gave them his files on Trump. They contained findings about "Trump's past dealings, including tax and bankruptcy problems, potential ties to organized crime, and numerous legal entanglements. They also revealed that Trump had an unusually high number of connections to Russians with questionable backgrounds."


title: Steele dossier

text: For months, Fusion GPS gathered information about Trump, focusing on his business and entertainment activities. When Trump became the presumptive nominee on May 3, 2016,<ref name="Johnson_5/4/2016"></ref> the conservative donor stopped funding the research on him.<ref name="Shane_Confessore_Rosenberg_1/12/2017"></ref><ref name="Borger_1/12/2017"></ref>


title: Steele dossier

text: Although the source of the Steele dossier's funding had already been reported correctly over a year before,<ref name="Robertson_2/7/2018"></ref> and the "Free Beacon" had issued a statement to this effect in October 2017, a February 2, 2018, story by the Associated Press (AP) contributed to confusion about its funding by stating that the dossier "was initially funded" by the "Washington Free Beacon", so the AP posted a correction the next day: "Though the former spy, Christopher Steele, was hired by a firm that was initially funded by the Washington Free Beacon, he did not begin work on the project until after Democratic groups had begun funding it."<ref name="Tucker_Jalonick_Day_2/3/2018_correction"></ref>


title: Steele dossier

text: By the spring of 2016, researchers at Fusion GPS had become so alarmed by what they had already learned about Trump that they felt the need "to do what they could to keep Trump out of the White House".


title: Steele dossier

text: The second operation of opposition research was indirectly funded by the DNC and the Clinton campaign, working through their attorney of record, Marc Elias of Perkins Coie.<ref name="Entous_Barrett_Helderman_10/24/2017"></ref> In an October 2017 letter, Perkins Coie general counsel Matthew Gehringer described how, in March 2016, Fusion GPS approached Perkins Coie and, knowing the Clinton campaign and the DNC were its clients, inquired whether its clients wished to pay Fusion GPS "to continue research regarding then-presidential candidate Donald Trump, research that Fusion GPS had conducted for one or more other clients during the Republican primary contest."<ref name="Watkins_Merica_Polantz_10/25/2017"></ref> In April 2016, Elias hired Fusion GPS to perform opposition research on Trump.


title: Steele dossier

text: Fusion GPS co-founder Glenn R. Simpson had some reservations, as he did not like the idea of helping Hillary Clinton. In an email, Simpson said "The only way I could see working for HRC is if it is against Trump."<ref name="Walsh_11/22/2019"></ref>


title: Steele dossier

text: In June 2016, as part of its work for Perkins Coie, Fusion GPS hired Orbis Business Intelligence, a private British intelligence firm, to look into connections between Trump and Russia. Orbis co-founder Christopher Steele, a retired British MI6 officer with expertise in Russian matters, was hired as a subcontractor to do the job.<ref name="Johnson_Kelly_1/9/2018"></ref> Prior to his work on the dossier, Steele had been a paid informant for the FBI<ref name="Winter_8/3/2018"></ref> for information unrelated to the Russia investigation.<ref name="Philips_2/24/2018"></ref>


title: Steele dossier

text: In total, Perkins Coie paid Fusion GPS $1.02million in fees and expenses, of which Fusion GPS paid Orbis $168,000 to produce the dossier.<ref name="Hosenball_11/1/2017"></ref> The DNC and Clinton campaign disclosed the total amount paid to Perkins Coie on campaign finance reports.<ref name="Johnson_9/10/2018"></ref> In March 2022, the Federal Election Commission (FEC) fined the DNC $105,000 and the Clinton campaign $8,000 for misreporting those fees and expenses as "legal services" and "legal and compliance consulting" rather than "opposition research".<ref name="Scott_3/30/2022"></ref><ref name="Cohen_3/30/2022"></ref>


title: Steele dossier

text: Orbis was hired between June and November 2016, and Steele produced 16 reports during that time, with a 17th report added in December. The reports were like "prepublication notes" based on information from Steele's sources, and were not released as a fully vetted and "finished news article".<ref name="Flegenheimer_1/8/2018"></ref> Steele believes 70–90 percent of the dossier is accurate, a view shared by Simpson.


title: Steele dossier

text: Steele delivered his reports individually to Fusion GPS as one- to three-page numbered reports. The first report, dated June 20, 2016, was sent to Washington by courier and hand-delivered to Fusion GPS. The names of the sources were redacted and replaced with descriptions to help Fusion judge their credibility.<ref name="Mayer_3/12/2018"></ref>


title: Steele dossier

text: Steele has said he soon found "troubling information indicating connections between Trump and the Russian government". According to his sources, "there was an established exchange of information between the Trump campaign and the Kremlin of mutual benefit". According to Harding, "Steele was shocked by the extent of collusion his sources were reporting", and told his friends: "For anyone who reads it, this is a life-changing experience."<ref name="Borger_11/15/2017"></ref> Steele felt what he had unearthed "was something of huge significance, way above party politics".<ref name="Blum_3/30/2017"></ref> American reporter Howard Blum described Steele's rationale for becoming a whistleblower: "The greater good trumps all other concerns."


title: Steele dossier

text: On his own initiative, Steele decided to also pass the information to British and American intelligence services because he believed the findings were a matter of national security for both countries.<ref name="Sengupta_1/13/2017"></ref> In 2018, Steele told a UK parliamentary investigation that Theresa May's British government covered up the evidence he provided them of Trump's Russian ties and took no actions, and that Boris Johnson suppressed a report about the intelligence in the dossier that was prepared by Parliament's Intelligence and Security Committee.<ref name="Sabbagh_6/22/2020"></ref> After long delay, the report was published on July 21, 2020.<ref name="Sabbagh_7/21/2020"></ref>


title: Steele dossier

text: According to Simpson's testimony, Steele, who enjoyed a good working reputation "for the knowledge he had developed over nearly 20 years working on Russia-related issues for British intelligence, approached the FBI because he was concerned that Trump, then a candidate, was being blackmailed by Russia,<ref name="Herb_Raju_Cohen_1/10/2018"></ref> and he became "very concerned about whether this represented a national security threat". Steele believed the intelligence community "needed urgently to know—if it didn't already—that the next possible U.S. president was potentially under the sway of Russia".<ref name="Simpson_Fritsch_11/26/2019"></ref>


title: Steele dossier

text: In early July 2016, Steele called seasoned FBI agent Michael Gaeta, who was stationed in Rome, and asked him to come to London so he could show him his findings. Because he was assigned to the U.S. embassy in Rome, Gaeta sought and was granted approval for the trip from Victoria Nuland, who was then the Assistant Secretary of State for European and Eurasian Affairs. When he arrived in London on July 5, 2016, he met with Steele at his office, and he was given a copy of Steele's first report, dated June 20, 2016 (Report 80). His reaction was "shock and horror". Alarmed by what he read, Gaeta remarked, "I have to show this to headquarters".


title: Steele dossier

text: As Nuland later shared, "In the middle of July, when he [Steele] was doing this other work and became concerned, he passed two to four pages of short points of what he was finding and our immediate reaction to that was, 'This is not in our purview.' This needs to go to the FBI if there is any concern here that one candidate or the election as a whole might be influenced by the Russian Federation. That's something for the FBI to investigate."<ref name="Tillett_2/4/2018"></ref> Shortly after, in July, the report was sent to an agent with expertise in criminal organizations and organized crime at the FBI's New York field office—essentially, the wrong person to handle a counterintelligence investigation.<ref name="Levine_9/18/2018"></ref> According to Nancy LeTourneau, political writer for the "Washington Monthly", the report "was languishing in the FBI's New York field office" for two months, and "was finally sent to the counterintelligence team investigating Russia at FBI headquarters in Washington, D.C." in mid-September 2016.


title: Steele dossier

text: In August 2016, the FBI asked Steele for "all information in his possession and for him to explain how the material had been gathered and to identify his sources". In October 2016, Steele "described the sources' access, but did not provide names" to the FBI. By August 22, 2017, Steele had provided them with the names of the sources for the allegations in the dossier.<ref name="Ross_Mosk_Schwartz_8/22/2017"></ref>


title: Steele dossier

text: Meanwhile, in the July to September time frame, according to "The Washington Post", CIA Director John Brennan had started an investigation with a secret task force "composed of several dozen analysts and officers from the CIA, the NSA and the FBI". At the same time, he was busy creating his own dossier of material documenting that "Russia was not only attempting to interfere in the 2016 election, they were doing so in order to elect Donald Trump... [T]he entire intelligence community was on alert about this situation at least two months before [the dossier] became part of the investigation." The "Steele dossier has so far proven to be fairly accurate", LeTourneau wrote.<ref name="LeTourneau_9/19/2018"></ref>


title: Steele dossier

text: In early August, Steele was summoned to Rome where he gave a full briefing to four American FBI officials about the report.<ref name="Kessler_1/9/2018"></ref> At that time, he handed over the June 20 and July 26 reports.


title: Steele dossier

text: During its intense questioning of Steele, the FBI mentioned their own discoveries of connections between the Trump campaign and Russia and asked Steele about Papadopoulos, but he said he knew nothing about him. The agents "raised the prospect of paying Steele to continue gathering intelligence after Election Day", but Steele "ultimately never received payment from the FBI for any 'dossier'-related information". In October 2022, during questioning from Special Counsel John Durham, Brian Auten, a supervisory counterintelligence analyst with the FBI, testified that, shortly before the 2016 election, the FBI offered Steele "up to $1 million" if he could corroborate allegations in the dossier, but that Steele could not do so.<ref name="Cohen_10/11/2022"></ref><ref name="Rizzo_10/12/2022"></ref> Steele has disputed this description: "And to correct the Danchenko trial record, we were not offered $1 million by the FBI to ‘prove up’ our Trump-Russia reporting. Rather, we were told there were substantial funds to resettle sources in the US if they were prepared to testify in public. Understandably they were not."<ref name="Steele_10/19/2022"></ref>


title: Steele dossier

text: The subsequent public release of the dossier stopped discussions between Steele and the FBI.<ref name="Hamburger_2/28/2017"></ref> Simpson testified that "Steele wasn't paid by the FBI, but was possibly reimbursed for a trip to Rome to meet with FBI officials."<ref name="Kiely_2/27/2018"></ref> According to Mayer, Steele asked the FBI to cover his travel expenses to Rome, but he received nothing. The Inspector General's report later confirmed that the FBI had initially offered to pay Steele $15,000 for his trip to Rome, but when the FBI dropped Steele as a confidential human source because he had shared information with a third party "in late October 2016" ("Mother Jones" magazine), the payment was halted. Peter Strzok reported that:


title: Steele dossier

text: In September, Steele met with Jonathan Winer, who was then the U.S. deputy assistant secretary of state for international law enforcement, whom he had known since 2009. In a 2018 editorial for "The Washington Post",<ref name="Winer_2/9/2018"></ref> Winer recounted that during their meeting in Washington, he was allowed to review Steele's reports, but not to keep a copy: "I prepared a two-page summary and shared it with [Victoria] Nuland, who indicated that, like me, she felt that the secretary of state needed to be made aware of this material," he wrote. Later in September, Winer discussed the report with Sidney Blumenthal, who revealed he had received similar information from Cody Shearer, a controversial political activist and former journalist who was close to the Clinton White House in the 1990s. Winer met with Steele again in late September and gave him a copy of Shearer's report, later known as the "second dossier".


title: Steele dossier

text: On September 19, 2016, Steele's handling agent sent six of Steele's reports (80, 94, 95, 100, 101, and 102) to the Crossfire Hurricane team, that had been operational since July 31, 2016. This was the first time they and their leader, Deputy Assistant Director of the FBI's Counterespionage Section, Peter Strzok, received any of Steele's reporting.<ref name="Levine_8/29/2018"></ref> Some reports referred to members of Trump's inner circle. After that point, he continued to share information with the FBI. The IG Report says this material "became an important part of the Crossfire Hurricane investigation and the FBI seeking FISA authority targeting one of the Crossfire Hurricane subjects, Carter Page."


title: Steele dossier

text: On October 28, 2016, days before the election, Comey notified Congress that the FBI had started looking into newly discovered Hillary Clinton emails. Simpson and Fritsch described their reaction: "Comey's bombshell prompted the Fusion partners to decide they needed to do what they could to expose the FBI's probe of Trump and Russia. It was Hail Mary time." The founders of Fusion GPS were very upset by a misleading November 1, 2016, "New York Times" article "published a week before the election with the headline: 'Investigating Donald Trump, FBI Sees No Clear Link to Russia'. In fact, Russia "was" meddling in the election to help Trump win, the U.S. intelligence community would later conclude..."


title: Steele dossier

text: Simpson later said that "Steele severed his contacts with [the] FBI before the election following public statements by the FBI that it had found no connection between the Trump campaign and Russia and concerns that [the FBI] was being 'manipulated for political ends by the Trump people'."<ref name="Jalonick_1/9/2018"></ref> Steele had become frustrated with the FBI, whom he believed failed to investigate his reports, choosing instead to focus on the investigation into Clinton's emails. According to "The Independent", Steele came to believe there was a "cabal" inside the FBI, particularly its New York field office linked to Trump advisor Rudy Giuliani, because it blocked any attempts to investigate the links between Trump and Russia.


title: Steele dossier

text: According to Fusion GPS's co-owners, Glenn Simpson and Peter Fritsch, they did not tell Steele who their ultimate clients were, only that Steele was "working for a law firm", and they "gave him no specific marching orders beyond this basic question: 'Why did Mr. Trump repeatedly seek to do deals in a notoriously corrupt police state that most serious investors shun?'" In testimony to the Senate Intelligence Committee, Simpson said that "it was Fusion GPS's idea to pursue overseas ties—that research was not directed by Perkins Coie, the DNC, or the Clinton Campaign".<ref name="Senate_8/18/2020"></ref>


title: Steele dossier

text: Jane Mayer reported that when the Clinton campaign "indirectly employed" Steele, Elias created a "legal barrier" by acting as a "firewall" between the campaign and Steele. Thus, any details were protected by attorney–client privilege and work-product privileges. "Fusion briefed only Elias on the reports, Simpson sent Elias nothing on paper—he was briefed orally", Mayer reported. In its application for a FISA warrant to survey Carter Page, the Department of Justice told the FISC that Simpson had not informed Steele of the motivation behind the research into Trump's ties with Russia. Steele testified to Congress that he did not know the Clinton campaign was the source of the payments "because he was retained by Fusion GPS".<ref name="Rocah_Goldman_McQuade_7/25/2018"></ref><ref name="Raju_Herb_10/26/2017"></ref> By "late July 2016", "several months" after signing the contract with Fusion GPS, Steele became aware that the DNC and the Clinton campaign were the ultimate clients.


title: Steele dossier

text: A spokesperson for the DNC said neither Tom Perez nor "the new leadership of the DNC were... involved in any decision-making regarding Fusion GPS, nor were they aware that Perkins Coie was working with the organization." A spokesperson for Perkins Coie said the campaign and the DNC were unaware Fusion GPS "had been hired to conduct the research". "The Washington Post" reported that it is not clear how much of the research Elias received from Fusion GPS he shared with the campaign and the DNC. It is also not clear who in those organizations knew about the roles of Fusion GPS and Steele, but one person "close to the matter" said the organizations were "not informed by the law firm of Fusion GPS's role". "The New York Times" revealed that earlier in 2017, "Mr. Elias had denied that he had possessed the dossier before the election."<ref name="Vogel_10/24/2017"></ref> The Clinton campaign did not know about Steele or that he was sharing his findings with the FBI, and "one top Clinton campaign official" told Jane Mayer that "If I'd known the F.B.I. was investigating Trump, I would have been shouting it from the rooftops!"


title: Steele dossier

text: The firewall was reportedly so effective even campaign principals John Podesta and Robby Mook did not know Steele was on the Democratic payroll until "Mother Jones" reported on the issue on October 31, 2016, in an article that did not name Steele. When the "Mother Jones" story broke, John Podesta, chairman of the Clinton campaign, said he was "stunned by the news that the FBI had launched a full-blown investigation into Trump, especially one that was informed by research underwritten by the Clinton campaign." Although they knew Perkins Coie had spent money for opposition research, neither Podesta nor campaign manager Robby Mook knew Steele was on the Democratic payroll. Mayer said they both maintain they "didn't read the dossier until "BuzzFeed" posted it online". She has also said that "the Clinton campaign never learned that Christopher Steele was on their payroll until it [the dossier] was in the press."<ref name="NPR_3/6/2018"></ref> "Far from a secret campaign weapon, Steele turned out to be a secret kept from the campaign." In their 2019 book, the founders of Fusion GPS wrote "...that no one from Fusion ever met or talked with Clinton and that she herself 'had no idea who they were'."


title: Steele dossier

text: Philip Bump wrote that the dossier "has never been shown to have informed the Clinton campaign's approach and that was not made public until shortly before Trump was inaugurated."<ref name="Bump_8/3/2020"></ref>


title: Steele dossier

text: On February 15, 2022, "The Washington Post" reported: "So far, there is no evidence that the Clinton campaign directly managed the Steele reporting or leaks about it to the media."<ref name="Kessler_2/15/2022"></ref>


title: Steele dossier

text: The founders of Fusion GPS have described how they did not hide the fact that they were researching Trump and Russia: "Fusion and Steele tried to alert U.S. law enforcement and the news media to the material they'd uncovered..." and their office became "something of a public reading room" for journalists seeking information. In September they arranged a private meeting between Steele and reporters from "The Washington Post", "The New York Times", "The New Yorker", ABC News, and other outlets. The results were disappointing, as none published any stories before the election.


title: Steele dossier

text: Jane Mayer has described how, in "late summer, Fusion set up a series of meetings, at the Tabard Inn, in Washington, between Steele and a handful of national-security reporters... Despite Steele's generally cool manner, he seemed distraught about the Russians' role in the election." Mayer attended one of the meetings. None of these news organizations ran any stories about the allegations at that time.


title: Steele dossier

text: Before the election, only two news sources mentioned allegations that came from dossier reports. Steele had been in contact with both authors. These were a September 23, 2016, Yahoo! News article by Michael Isikoff that focused on Carter Page, and an article by David Corn on October 31, 2016, a week before the election, in "Mother Jones" magazine.


title: Steele dossier

text: At the time, the FBI had assumed that the source for the Yahoo! article was someone, other than Steele, "who had received a copy of the dossier". The 2019 DOJ report by Michael Horowitz criticized the FBI for failing to ask Steele if he had a role in that Yahoo! article.<ref name="Savage_Goldman_4/12/2021"></ref>


title: Steele dossier

text: By the third quarter of 2016, many news organizations knew about the existence of the dossier that had been described as an "open secret" among journalists, but chose not to publish information they could not confirm.<ref name="Ember_Grynbaum_1/10/2017"></ref> "Mother Jones" was the first to report the existence of the dossier and that it was exclusively funded by Democrats.<ref name="Blake_10/25/2017"></ref><ref name="O'Brien_10/26/2017"></ref>


title: Steele dossier

text: By October 2016, Steele had compiled 33 pages (16 reports), and he then passed on what he had discovered to David Corn, a reporter from "Mother Jones" magazine. On October 31, 2016, a week before the election, "Mother Jones" reported that a former intelligence officer, whom they did not name, had produced a report based on Russian sources and turned it over to the FBI.<ref name="Corn_10/31/2016"></ref> The article disclosed some of the dossier's allegations:


title: Steele dossier

text: After Trump's election on November 8, 2016, the Democratic client stopped paying for the investigation, but Steele continued working on the dossier for Fusion GPS. According to "The Independent", at that time, Simpson "reportedly spent his own money to continue the investigation".<ref name="Sampathkumar_8/23/2017"></ref> According to "The New York Times", after the election, Steele's dossier became one of Washington's "worst-kept secrets", and journalists worked to verify the allegations.


title: Steele dossier

text: On November 18, 2016, Republican Senator John McCain, who had been informed about the alleged links between the Kremlin and Trump, met with former British ambassador to Moscow Sir Andrew Wood at the Halifax International Security Forum in Canada. Wood told McCain about the existence of the collected materials about Trump,<ref name="Edwards_1/11/2017"></ref> and also vouched for Steele's professionalism and integrity.<ref name="Sengupta_1/12/2017"></ref>


title: Steele dossier

text: According to Simpson's August 22, 2017, testimony to the Senate Judiciary Committee, Steele and David J. Kramer, a longtime McCain aide and former U.S. State Department official working at Arizona State University, met each other at the Halifax forum and discussed the dossier. Kramer told Steele that McCain wanted to "ask questions about it at the FBI... All we sort of wanted was for the government to do its job and we were concerned about whether the information that we provided previously had ever, you know, risen to the leadership level of the FBI."


title: Steele dossier

text: Steele had agreed with Fusion GPS to deliver a hard copy of all 16 reports to McCain, that McCain received in early December from Kramer. On December 9, McCain met personally with FBI Director James Comey and gave him a copy of the dossier.<ref name="Resnick_5/9/2018"></ref> On March 25, 2019, Senator Lindsey Graham, a close friend of McCain's, contradicted Trump's attacks against McCain and revealed that when McCain showed him the dossier, he had encouraged McCain to give the dossier to the FBI and that McCain acted appropriately.<ref name="Sonmez_3/25/2019"></ref> Graham described how he confronted Trump: "Senator McCain deserves better. There were some McCain people who took a piece of garbage and tried to go after Trump after the election. But I told the president it was not John McCain."<ref name="Dennis_3/25/2019"></ref> Comey later confirmed that counterintelligence investigations were under way into possible links between Trump associates and Moscow.


title: Steele dossier

text: After delivering his 16 reports to McCain, Steele received more information and composed the two-page "December memo", dated December 13. It mostly contained allegations against Trump's personal attorney, Michael Cohen, that Cohen later denied. In an April 2017 court filing, Steele revealed previously unreported information that he had given a copy of his last report to a "senior UK government national security official acting in his official capacity, on a confidential basis in hard copy form", because it "had implications for the national security of the US and the UK".


title: Steele dossier

text: Both Simpson and Steele have denied providing the dossier to "BuzzFeed". Unsealed documents from the discovery process in Russian entrepreneur Aleksej Gubarev's defamation lawsuit revealed that David Kramer, an associate of John McCain, gave the dossier to "BuzzFeed" "in December 2016, weeks after the election",<ref name="Cohen_Collier_3/16/2019"></ref><ref name="Gross_Simpson_Fritsch_11/26/2019"> "Fresh Air"</ref> in what Fritsch has called an "ill-advised" Hail Mary pass. It was never intended that the dossier be public because it was unfinished raw intelligence and could have "compromised sources and methods". Steele was also worried about the safety of his source network, and he expressed concern that if Trump won the election, the new FBI director, and other agency heads appointed by Trump, might be more loyal to Trump "and could decide to take action against Steele and his source network".


title: Steele dossier

text: On January 5, 2017, the chiefs of four U.S. intelligence agencies briefed President Barack Obama and Vice President Joe Biden about the Russian interference in the election and the existence of the dossier and its allegations.<ref name="Nowicki_1/13/2017"></ref><ref name="AOL_1/12/2017"></ref>


title: Steele dossier

text: On the afternoon of January 6, 2017, President-elect Trump and his transition team received a similar briefing in Trump Tower.<ref name="Kosoff_7/20/2017"></ref> All four of the top intelligence chiefs met with Trump and his transition team. They were Director of National Intelligence James Clapper, FBI Director James Comey, CIA Director John Brennan, and NSA Director Admiral Mike Rogers. They informed Trump of the Russian election interference, and Comey told them of "a piece of Steele's reporting that indicated Russia had files of derogatory information on both Clinton and the President-elect".


title: Steele dossier

text: Then, according to a pre-arranged plan, Brennan, Clapper, and Rogers left, and Comey then asked to speak with Trump alone. Comey then informed Trump of the dossier and its allegations about salacious tapes held by the Russians. Comey later reported he was very nervous. The previous day, the secretary of the Department of Homeland Security told Comey to "be very careful", "choose your words carefully", and then "get outta there". Trump became very defensive, and Comey described the meeting as "really weird".<ref name="Chan_4/16/2018"></ref> Trump later expressed that he felt James Comey was trying to blackmail him at the meeting in Trump Tower, held two weeks before the inauguration. In April 2018, Comey said he did not inform Trump the dossier was partly funded by Democrats because that "wasn't necessary for my goal, which was to alert him that we had this information".<ref name="Stracqualursi_4/13/2018"></ref><ref name="McCarthy_4/13/2018"></ref>


title: Steele dossier

text: The Mueller Report, published on April 18, 2019, contains a footnote that suggests that Trump may have heard that Russia had incriminating tapes of his behavior. On October 30, 2016, Michael Cohen had received a text from Giorgi Rtskhiladze reporting that he had successfully stopped the "flow of tapes from Russia". Rtskhiladze told investigators that these were compromising tapes of Trump, and Cohen told investigators he had spoken to Trump about the issue. Rtskhiladze later told investigators "he was told the tapes were fake, but he did not communicate that to Cohen".


title: Steele dossier

text: On December 14, 2018, the FBI released a document called "Annex A", that was "part of [the] Russia dossier summary" used to brief Trump and Obama.<ref name="Gerstein_Cheney_12/14/2018"></ref> The FBI withheld parts of the synopsis on the grounds that it remained classified and "because it pertains to ongoing investigations or court proceedings, originated with a confidential source or describes confidential investigative techniques or procedures".


title: Steele dossier

text: Two days after the publication of the dossier on January 10, 2017, James Clapper issued a statement describing the leaks in the press about their Trump Tower meeting with Trump as damaging to U.S. national security.<ref name="Hartmann_1/12/2017"></ref> The statement also included the non-committal wording that the U.S. intelligence community "has not made any judgment that the information in this document is reliable, and we did not rely upon it in any way for our conclusions". This contradicted Trump's previous claim that Clapper had said the information in the dossier was false; Clapper's statement actually said the intelligence community had made no judgment on the truth of the information.<ref name="Weisman_Steinhauer_1/12/2017"></ref>


title: Steele dossier

text: James Comey had disagreed with Clapper's wording, but Clapper's wording remained unchanged. Comey later told the Office of the Inspector General of his concerns at that time, because he believed the dossier to be more reliable than indicated in Clapper's non-committal statement:


title: Steele dossier

text: On January 10, 2017, CNN reported that classified documents presented to Obama and Trump the previous week included allegations that Russian operatives possess "compromising personal and financial information" about Trump. CNN said it would not publish specific details on the reports because it had not "independently corroborated the specific allegations".<ref name="Perez_Sciutto_Tapper_Bernstein_1/10/2017"></ref><ref name="Wemple_1/11/2017"></ref> Following the CNN report, "BuzzFeed" published a draft dossier that it said was the basis for the briefing, including unverified claims that Russian operatives had collected "embarrassing material" involving Trump that could be used to blackmail him. "BuzzFeed" said the information included "specific, unverified, and potentially unverifiable allegations of contact between Trump aides and Russian operatives".<ref name="Bensinger_1/10/2017"></ref><ref name="Mak_Suebsaeng_Weiss_ 1/10/2017"></ref><ref name="Shane_Goldman_Rosenberg_1/10/2017"></ref>


title: Steele dossier

text: "The New York Times" commented: "Mr. Steele has made clear to associates that he always considered the dossier to be raw intelligence—not established facts, but a starting point for further investigation."<ref name="Shane_Goldman_Rosenberg_4/19/2019"></ref> Judge Craig Karsnitz described the purpose of the dossier: "An intelligence report is simply a report of information potentially relevant to an investigation. It can take many forms, be true or false, and can be used as opposition research and an intelligence report." Christopher Burrows, co-founder of Orbis Business Intelligence, does not consider it a "dossier", but "information that is referred to as 'raw intelligence' in intelligence circles... a mixture of knowledge, rumor and hearsay... [A]n intelligence agency would enrich the findings with data, test probabilities and write analyses. It's an elaborate process. But Steele is not an intelligence agency."


title: Steele dossier

text: "BuzzFeed" was harshly criticized by several mainstream media outlets for releasing the draft dossier without verifying its allegations.<ref name="Bump_1/11/2017"></ref><ref name="Zurawik_1/11/2017"></ref> "Washington Post" columnist Margaret Sullivan called it "scurrilous allegations dressed up as an intelligence report meant to damage Donald Trump",<ref name="Sullivan_1/11/2017"></ref> while "The New York Times" noted that the publication sparked a debate centering on the use of unsubstantiated information from anonymous sources.<ref name="Smith_1/23/2017"></ref> "BuzzFeed"s executive staff said the materials were newsworthy because they were "in wide circulation at the highest levels of American government and media" and argued that this justified public release.<ref name="Graham_1/11/2017"></ref> A judge in the Superior Court of the District of Columbia agreed with this reasoning when he threw out a libel suit against Steele and Orbis Business Intelligence.