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The final mainstream common issue affects all EA888 generations. The cooling system is mainly a problem free system, with the exception of the plastic thermostat unit, these are very commonly known to be prone to leaks, with no specific part of the housing known to leak. On the EA888, the thermostat unit also includes the coolant pump, on the Generation 2 & 3 the coolant pump is still part of the thermostat, however is available separately. The coolant pump / thermostat unit is located under the intake manifold regardless of generation, model year or application. The thermostat side is joined by a plastic union directly to the engine oil cooler, which in turn is mounted to and is an integral part of the ancillary / alternator bracket (also includes the oil filter housing in all generations and applications). The coolant pump is driven by the intake side balance shaft, on the flywheel side of the engine. The rectification is to renew the thermostat unit with a modified unit, and if needed in later models, the coolant pump if necessary. However these newer units are still known to leak. There have currently been no further modifications to the design of this to combat the issues by Volkswagen Group.
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Depending on the species and strain of rodent used, at least 12 different behaviors have been noted in the hot plate test, including sniffing, grooming, stamping of the legs, freezing, licking, leaning and jumping (Espejo and Mir, 1993). Some of these behaviors can be sensitive to analgesics, although differences are observed depending on the type of behavior quantified. For example, paw licking is diminished by opioids but not other analgesics, while other behaviors can also be affected by other classes of analgesics (Ankier, 1974; Hunskaar et al., 1985). Data quality is usually improved if the time to occurrence of any behavior, rather than specific behavior types, is recorded, and if lower temperatures are used (Carter, 1991; Plone et al., 1996). It is plausible that differences in behavior may relate to the type of sensory fiber activated. In anesthetized rats, steep temperature gradients and high skin temperatures are associated with activation of Aδ fibers, while slower heating and lower temperatures lead to firing of C fibers (Yeomans and Proudfit, 1994, 1996; Yeomans et al., 1996).
Temperature preference assays. (A) Two-temperature choice assay. Rodents are allowed to freely move between a reference plate (neutral temperature) and test plate. The time spent on the test plate relative to the reference plate is measured over a set period of time. (B) Continuous temperature gradient assay. Rodents are allowed to freely move along a liner or circular surface with a temperature gradient. The time taken to settle in a temperature zone and/or the temperature of the chosen zone is recorded.
Methods used to assess non-stimulus evoked pain behaviors in rodents. (A) Grimace scales. Facial expression is subjectively scored for severity of pain based on five facial features (ear position, eye closing, cheek bulging, whisker position, and nose bulging). (B) Burrowing assay. A burrow is placed in the cage of a rodent filled with a suitable substrate (such as food pellets, sand, or marbles). The amount of substrate displaced over a set period of time is recorded. Pain in rodents is associated with decreased burrowing behaviors. (C) Weight bearing (incapacitance test). The rodent is placed in an inclined holder with the hind paws resting on two separate pressure sensors. Weight distribution between the hind paws is recorded. (D) Gait analysis (Catwalk XT, Noldus). In this assay rodents walk freely across an enclosed elevated glass floor. A camera below records the paw prints, which are illuminated by internally reflected light in the glass. A number of parameters are automatically analyzed by the software, including paw intensity, print area, stance phase duration (time spent on paw) and swing phase duration (time spent off paw). (E) Behavioral Spectrometer (Behavioral Instruments). Rodents are placed in an enclosed box with a camera, accelerometer and wall-mounted photobeams for a set period of time. The software records the duration of different behavior types, including movement, grooming and rearing behaviors.
The Advanced Dynamic Weight Bearing apparatus (Bioseb) was developed as a modification from static weight bearing or incapacitance tests and computes weight bearing for each of the front and rear paws, weight ratio and paw surface area in freely moving animals (Griffioen et al., 2015). The dynamic weight-bearing test is able to detect reduced weight bearing behaviors of the affected hind limb in multiple pain models, including CFA-induced inflammation, chronic constriction injury, bone cancer pain and antigen-induced arthritis (Tetreault et al., 2011; Robinson et al., 2012; Quadros et al., 2015). Similarly, it overcomes some of the experimental difficulties of static weight bearing analysis, albeit no information on gait can be obtained. 2b1af7f3a8