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Most of the rats achieved the pellets per session criterion after 10 to 15 sessions. Presses on the inactive lever were recorded but had no programmed consequence. Again, a minimum of responses per session on the active lever was required for the advancement to the next schedule; most of the rats required only one to two FR3 and FR5 schedule s to achieve this level.

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The FR5 schedule was followed by the progressive ratio PR schedule during which the cost of a reward is progressively increased for each following reward in order to determine the amount of work the rat is willing to put into obtaining the reward. The response requirement increased according to the following equation: The PR session ended when the rat had failed to earn a reward within 60 minutes. The break point was defined as the final completed ratio before the session ended.

In most cases, responding stabilized within five to seven sessions. The sessions lasted, on average, 75 minutes, although all the rats stayed in the operant boxes until minutes to allow for all the sessions to end. The rats were subsequently transferred to home cages for a one hour free-feeding chow intake measurement. At the end of training and prior to testing, the rats were returned to an ad libitum feeding schedule.

Experimental design All of the rats received intraperitoneal IP or in a separate group of rats, third ventricle third ICV injections early in the light cycle for ghrelin tests and late in the light cycle for ghrelin antagonist experiments 20 minutes prior to the start of operant testing.

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All conditions were separated by a minimum of 48 hours and run in a counterbalanced manner each rat received all conditions on separate testing days. The selected IP ghrelin dose has been shown previously to induce a feeding response in rats Wren et al.

Subsequent to operant testing, the rats were allowed free access to chow, and chow intake was measured after a one-hour period.

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Next, in a separate group of rats, we examined responses after targeted CNS drug delivery after three conditions as follows: The selected doses of ghrelin has previously been shown to elicit feeding responses Nakazato et al. For both the ICV and the IP ghrelin studies, lever-pressing experiments were performed in the satiated state i.

Also, in both studies, subsequent to operant testing, the rats were allowed free access to chow, and chow intake was measured after a one-hour period. The JMV doses were selected based on Jerlhag et al.

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Subsequent to operant testing, the rats were allowed free access to chow. To assess the effects of direct acute central antagonist action, in a separate group of rats, operant behavior was examined after the following three conditions: Subsequent to operant testing, the rats were allowed free access to chow and chow intake was measured after a one-hour period and also at 24 hours after the initial injection.

Studies with the GHS-R1A antagonist, in contrast to those performed with ghrelin see earlierwere performed on the rats after a hour food restriction prior to the injections in order to ensure high levels of endogenous circulating ghrelin Cummings et al.

The acetylcholine-related genes were: Drug administration and tissue dissection The catheter and the osmotic pump were filled with acetylated human ghrelin gift from Rose Pharma, Copenhagen, Denmark solution 8. Fourteen days after implantation of the minipumps, the rats were killed by decapitation. All the cDNA-reactions were run in triplicate. They were designed with TaqMan probe and primer sets for target genes chosen from an on-line catalogue Applied Biosystems.

Thermal cycling conditions were: Glyceraldehydephosphate dehydrogenase was used as reference gene. Statistics All behavioral parameters were analyzed by analysis of variance followed by post hoc Tukey test or t-tests as appropriate. Specifically, to determine the role of peripheral ghrelin administration on sucrose reward efficacy, we examined sucrose self-administration in a progressive response schedule in the rats 20 minutes after IP injection of vehicle or ghrelin.

All measures of operant behavior were significantly increased in the rats after acute peripheral ghrelin injection: The literature primarily supports a central site of action for ghrelin's orexigenic effect. However, GHS-R1A is also expressed outside of the CNS in sites relevant for food intake control, for example, on the vagus nerve; therefore, it can not be ruled out that part of the observed effects of IP ghrelin are mediated by those peripheral receptors.

Therefore, in order to determine a direct CNS effect of ghrelin on sucrose reward efficacy, we performed a parallel study in which vehicle or ghrelin were administered by third ventricle injection, also 20 minutes prior to the operant paradigm. Consistent with a central site of effect hypothesis, acute ICV ghrelin injection to rats both 0. The time course of the active lever responses in the ICV ghrelin study revealed that while the effect emerged slowly during the and minute time points, it reached significance at 60 minutes [active lever: Either way--honorifics or Maude and Alice--the two never let the tiny redwood cottage, which eventually sagged onto the garage next door, cramp their business.

Bythere were so many children and houses lined up for tintype portraits that Wilson opened a studio of her own on Kingsley Ave. Meanwhile, Kelly's photographs of fog prompted some of New York's most loquacious art critics to reach new heights of adjectival overload praising the "lyrical vein" and "delicate gauze of the mist particles" in Kelly's work. The four other houses are Embarcadero Road, Cowper St. Although some of these houses are regularly included in PAST's Saturday morning tours of historic Palo Alto, the holiday tour is the only time the public is allowed inside the houses.

Tickets may be purchased the day of the tour.

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Here, briefly, are a few of the historical and architectural highlights of the tour. It isn't easy blending turrets, triple gables and gambrel roof-wings with Craftsman style and ending up with a house that was long considered "the most attractive home in Palo Alto.

Smith, who built the house at Cowper St. Although gone now, the property once included a barn on Byron Street and a full-scale dairy wing.

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The dairy wing, alas, is now an apartment. Built "before, during and after the earthquake," the house has grown from two stories to three, from eight rooms to Although the back-yard beehives and chicken coops are long gone, part of the old aviary remains, as does an old wisteria vine that won't quit.

The house is pure Spanish Revival, from the red tile roofs to the wrought iron grillwork, truss-beam ceilings, tile floors, paved terrace and walled garden. Still known as the Pettigrew House, the house was built for businessman Perce Pettigrew. Although considered one of the finest examples of Spanish Revival architecture in the city, the house did not instantly engender affection among neighbors.