Partial reversal of anticholinergic amnesia by choline chloride

Twenty young adult males were given tests of memory and mood after receiving choline chloride plus scopolamine, placebo plus scopolamine, and two placebos on separate days in randomized order. Scopolamine caused a marked impairment in performance on a verbal learning task. Pretreatment with choline caused a small but significant decrease in this impairment and partially reversed the mood changes produced by scopolamine. Although several previous studies have failed to demonstrate that choline has a significant effect on cognition when given alone, the present results indicate that choline can enhance cognition under conditions of cholinergic blockade. This is consistent with preclinical data indicating that cholinergic neurons are most sensitive to precursor availability when they are firing rapidly.     

Rehmannia glutinosa ameliorates scopolamine-induced learning and memory impairment in rats

Many studies have shown that the steamed root of Rehmannia glutinosa (SRG), which is widely used in the treatment of various neurodegenerative diseases in the context of Korean traditional medicine, is effective for improving cognitive and memory impairments. The purpose of this study was to examine whether SRG extracts improved memory defects caused by administering scopolamine (SCO) into the brains of rats. The effects of SRG on the acetylcholinergic system and proinflammatory cytokines in the hippocampus were also investigated. Male rats were administered daily doses of SRG (50, 100, and 200 mg/kg, i.p.) for 14 days, 1 h before scopolamine injection (2 mg/kg, i.p.). After inducing cognitive impairment via scopolamine administration, we conducted a passive avoidance test (PAT) and the Morris water maze (MWM) test as behavioral assessments. Changes in cholinergic system reactivity were also examined by measuring the immunoreactive neurons of choline acetyltransferase (ChAT) and the reactivity of acetylcholinesterase (AchE) in the hippocampus. Daily administration of SRG improved memory impairment according to the PAT, and reduced the escape latency for finding the platform in the MWM. The administration of SRG consistently significantly alleviated memory-associated decreases in cholinergic immunoreactivity and decreased interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) mRNA expression in the hippocampus. The results demonstrated that SRG had a significant neuroprotective effect against the neuronal impairment and memory dysfunction caused by scopolamine in rats. These results suggest that SRG may be useful for improving cognitive functioning by stimulating cholinergic enzyme activities and alleviating inflammatory responses.     

吗啡和胆碱能拮抗剂联合给药对小鼠Y迷宫空间记忆提取及活动性的影响(英文）

吗啡和胆碱能系统的相互作用已在多项研究中提到,本实验想查明吗啡是否能和胆碱能拮抗剂、东莨菪碱以及阿托品共同作用对小鼠的Y迷宫空间识别记忆提取产生影响。采用测试前腹腔给药的方法,选用3种剂量的吗啡(5、1.5、0.5mg/kg),两种剂量的东莨菪碱(1、0.1mg/kg),以及两种剂量的阿托品(0.5、0.1mg/kg),剂量由高到低相配对作为联合给药的手段。其结果表明:1)0.5mg/kg低剂量吗啡与0.1mg/kg低剂量的东莨菪碱,或与0.1mg/kg低剂量的阿托品联合给药的小鼠,在记忆提取测试中,空间探查行为(各臂停留时间百分比)对新异臂没有偏好,而新奇探索行为(各臂访问次数百分比)仍保持了对新异臂的偏好,而相应剂量药物单独给药的小鼠记忆提取均没有被损害;2)吗啡能和东莨菪碱相互作用使小鼠的活动性显著增强。暗示吗啡和胆碱能拮抗剂对小鼠空间记忆提取的破坏存在一定程度的相互作用。     

Neuropsychological effects of marijuana

This study assigned 81 non-naïve subjects, divided into low- and high-dose groups, to four experimental conditions (marijuana/marijuana, marijuana/placebo, placebo/marijuana and placebo/placebo) for two sessions separated by about one week. The low dose was 4.8 mg. Δ⁹-THC followed by 2.4 mg. one hour later. The high dose was 9.1 mg. followed by 4.5 mg. one hour later. A battery of neuropsychological tests was administered.The low dose produced generalized impairment of all mental processes (concept formation, memory, tactile form discrimination and motor function) and the effect was generalized to all modalities. The high dose resulted in more extensive impairment, again generalized.The drug effects noted were explained in terms of generalized impairment of central integrative processes. The effects of marijuana on learning as well as memory were explained in terms of impaired output (recall), but the impairment was transient.     

Antinociceptive effects of morphine-6-glucuronide in homozygous MDR1a P-glycoprotein knockout and in wildtype mice in the hotplate test

Morphine-6-glucuronide (M6G), a major metabolite of morphine with agonist opioid-receptor activity, was reported to be a substrate of P-glycoprotein (P-gp). Inhibition of P-gp may thus result in higher brain uptake of M6G. The goal of this observer-blinded, placebo controlled study, was to compare the antinociceptive effects of M6G in homozygous P-gp knockout (mdr1a(-/-)) and wildtype (mdr1a(+/+)) mice. M6G was injected intraperitoneally as a single dose of 0, 0.5, 1, 2.5, 5, and 10 mg/kg. Eight P-gp knockout and eight wildtype mice were studied per dose. A hot plate test was performed before and 5, 15, 30, 60, 90, 120, and 150 min after M6G administration. Plasma-concentrations of M6G, morphine, and morphine-3-glucuronide (M3G) were measured after intraperitoneal injection of 5 mg/kg M6G in another 14 P-gp knockout and 14 wildtype mice. No difference neither in the dose response relationship, nor in the time course of response latency times were observed between P-gp knockout and wildtype mice. However, latency times increased with higher doses of M6G, with antinociception significantly different from placebo at a M6G dose of 5 and 10 mg/kg. P-gp knockout mice tended to have higher plasma concentrations than the wildtype. However, plasma concentrations widely overlapped between groups and therefore no statistical significant group difference could be detected. We conclude that despite reported doubling of M6G brain uptake, absence of mdr1a coded P-gp does not enhance antinociceptive effects of M6G in the hotplate test after acute single-dose administration in mdr1a(-/-) knockout mice.     

The interaction between the cholinergic and dopaminergic system in learning and memory process in rats.

In normal rats, muscarinic acetylcholine receptors (mAChRs) have a facilitating role on both short-term and long-term memory tested by Y-maze task and multi-trial passive avoidance test, respectively, since scopolamine, a specific mAChRs antagonist, impairs both types of memory. A low dose of nicotine (0.3 mg/kg b.w., i.p.), a specific nicotinic acetylcholine receptors (nAChRs) agonist, administered once caused a significant facilitating effect on short-term memory. A higher dose of nicotine (3 mg/kg b.w., i.p.) administered 5 consecutively days had about the same facilitating effect on short- and long-term memory without affecting information acquisition. In rats, having mAChRs and nAChRs blocked by means of scopolamine and chlorisondamine respectively, a low dose of nicotine administered once caused a significant improvement of long-term memory deficits without affecting significantly short-term memory. A higher dose of nicotine administered 5 consecutive days in rats with a double blockade of cholinergic receptors had the same ameliorating effect on long-term memory deficits as low dose. Our data suggest that the antiamnesic effect of nicotine can result from an action at nicotinic receptors subtypes not blocked by chlorisondamine or at nonnicotinic receptors.     

东莨菪碱慢性给药大鼠作为老龄相关记忆损害模型的探索

目的对东莨菪碱慢性给药大鼠能否作为老龄相关记忆损害模型进行探索。方法14只1月龄SD大鼠随机分为对照组和东莨菪碱模型组。东莨菪碱模型组大鼠皮下注射东莨菪碱2mg kg,2次日,正常对照组予等量生理盐水,连续21d。然后利用Morris水迷宫(MWM)参照记忆试验进行行为学测试;神经元的特殊染色及电子显微镜技术,观察大鼠海马CA1、CA3区锥体细胞数、超微结构的改变以及突触可塑性变化。结果东莨菪碱组大鼠隐匿平台搜索实验成绩有一定损害;两组大鼠空间探索次数差异无显著性(P>0.05)。两组间海马CA1、CA3区锥体细胞数差异无显著性(P>0.05)。两组大鼠锥体细胞胞体超微结构无差异,但两组大鼠CA1区神经元突触超微结构有轻微变化。结论东莨菪碱慢性给药对大鼠学习记忆能力有一定损害,但对长时记忆无明显影响;对海马神经元结构无明显损害,对神经元突触可塑性有轻微影响。此种动物模型可能不是理想的老年性痴呆或老年相关记忆损害模型。     

灵芝对小鼠空间分辨学习与记忆的影响

本文用Y-型迷宫法测试小鼠空间分辨行为。实验结果表明,每日ig灵芝2.58/kg共7d,有明显促进学习的作用。每日ig灵芝2.5g/kg共7d或ig灵芝5g/kg共7d都能显著地拮坑东莨菪碱所致学习障碍的作用。此外,学习训练后立即ig灵芝2.5g/kg或ig灵芝5g/kg也有明显地改善东莨菪碱损害记忆巩固的作用。     

Effect of a divided caffeine dose on endurance cycling performance, postexercise urinary caffeine concentration, and plasma paraxanthine.

This study compared the effects of a single and divided dose of caffeine on endurance performance and on postexercise urinary caffeine and plasma paraxanthine concentrations. Nine male cyclists and triathletes cycled for 90 min at 68% of maximal oxygen uptake, followed by a self-paced time trial (work equivalent to 80% of maximal oxygen uptake workload over 30 min) with three randomized, balanced, and double-blind interventions: 1) placebo 60 min before and 45 min into exercise (PP); 2) single caffeine dose (6 mg/kg) 60 min before exercise and placebo 45 min into exercise (CP); and 3) divided caffeine dose (3 mg/kg) 60 min before and 45 min into exercise (CC). Time trial performance was unchanged with caffeine ingestion (P = 0.08), but it tended to be faster in the caffeine trials (CP: 24.2 min and CC: 23.4 min) compared with placebo (PP: 28.3 min). Postexercise urinary caffeine concentration was significantly lower in CC (3.8 micro g/ml) compared with CP (6.8 micro g/ml). Plasma paraxanthine increased in a dose-dependent fashion and did not peak during exercise. In conclusion, dividing a caffeine dose provides no ergogenic effect over a bolus dose but reduces postexercise urinary concentration.     

Plasma concentrations of choline in man following choline chloride

Plasma choline levels were measured in patients being treated with choline chloride for movement disorders. Following single doses of 5 g given orally in aqueous solution, plasma concentrations rose to a peak within four hours and then rapidly declined. The degree of increase was variable both between and within patients. During chronic treatment, plasma choline concentrations tended to rise as the dose increased, although the relationship was not strong. The highest concentrations attained by patients were always at a dose of 16 or 20 g daily. Following chronic treatment, the disappearance of choline from plasma was rapid, with most patients reaching baseline by four days. Choline chloride is generally given in four divided doses, which seems reasonable in the early stages of treatment. Most therapeutic effect is seen when patients are treated with daily doses in the 12 to 20 g range, doses likely to produce substantial increases in plasma choline concentration. However, the relationship of plasma choline concentration to clinical efficacy may be tenuous. Following discontinuation of treatment, clinical improvement tends to persist long after plasma choline has returned to baseline concentrations.     

A multicenter,randomized, placebo controlled,multiple-dose,safety and pharmacokinetic study of AIT-082 (Neotrofin) in mild Alzheimer's disease patients

A phase 1, randomized, double-blind, placebo-controlled, dose escalation study of the purine derivative, AIT-082 (Neotrofin, NeoTherapeutics) was conducted in mild Alzheimer's disease (AD) patients to evaluate multiple-dose safety, tolerability, and pharmacokinetics. Possible short-term effects of AIT-082 on cognition and memory were preliminarily investigated. AIT-082 is currently being developed as a potential treatment for Alzheimer's disease and other neurological disorders. Pre-clinical studies indicate that AIT-082 has memory enhancing properties, stimulates neuritogenesis and the production of neurotrophic factors. Patients received an oral dose of AIT-082 or placebo daily for one week. Thirty-six AD patients were divided into three dose cohorts; each dose cohort consisted of twelve patients with 8 patients randomized to AIT-082 and 4 to placebo. The 3 doses of AIT-082 evaluated in this study were 100 mg/day, 500 mg/day, and 2,000 mg/day. There were no serious adverse events at any dose and the drug was well tolerated without significant side effects. AIT-082 was orally and rapidly absorbed, resulting in peak serum concentrations within 2 hours with an elimination half-life of approximately 20 hours. Higher doses resulted in corresponding increases in peak concentrations and areas under the curve (AUC). There was an approximate 2-fold accumulation in AIT-082 with daily dosing (as reflected by the AUC) at steady state. There were no significant differences by treatment arm on the clinical or neuropsychological evaluations. AIT-082 was rapidly absorbed by the oral route with a half-life suitable for dosing once or twice daily. No problems with tolerability or safety were found. AIT-082 appears suitable for testing in larger clinical trials for the treatment of AD and other neurologic disorders.     

Metabolism of cytidine (5′)-diphosphocholine (cdp-choline) following oral and intravenous administration to the human and the rat

We examined the effects of cytidine (5′)-diphosphocholine (CDP-choline) on plasma levels of cytidine, choline, and unchanged CDP-choline among normal volunteers receiving the substance orally or intravenously, and rats receiving it intravenously. Two hours after a single oral dose (2g), plasma choline levels were increased by 48% and plasma cytidine by 136%. Among subjects receiving three doses (2g each) at two-hour intervals, plasma choline peaked (30% over baseline) 4 h after the initial CDP-choline dose, while plasma cytidine levels continued to increase for at lest 6 h, at which time they were five times basal levels (P < 0.01). Intravenously-administered CDP-choline was rapidly hydrolysed, in both the human and the rat. In humans given the CDP-choline by infusion over 30 min, plasma CDP-choline fell to undetectable levels almost immediately after the end of the infusion period; plasma choline and cytidine peaked at that time, but their concentrations remained significantly elevated for at least 6 h. In rats given a bolus injection of CDP-choline, five minutes earlier, the unchanged compound was also undetectable in plasma, while plasma cytidine levels increased markedly and remained elevated for at least 60 min. These observations show that CDP-choline is converted to at least two major circulating metabolites, choline and cytidine. Since both of these compounds are used in the biosynthesis of phosphatidylcholine, both may be involved in the long-term effects of the CDP-choline.     

Use of a new steroid (Org OD 14) in the climacteric syndrome

A double-blind study was carried out in 60 women with climacteric symptoms: 30 women were given Org OD 14 (2.5 mg) and 30 were given a placebo to be taken daily for six weeks. The effects of the medication on the climacteric symptoms, the subjective sensations, the plasma FSH levels and endometrial histology were studied. In the treated group compared with the control group the relief or improvement of the following climacteric symptoms were recorded: perspiration, palpitations, irritability and backache. A favourable effect on the subjective sensations was noted in both groups, although no significant difference for the group which received Org OD 14 was found. At the end of the treatment with Org OD 14, the FSH levels were found to be greatly reduced in comparison with the basal values; this, however, was not the case with the placebo group. With regard to endometrial histology, no sign of hyperplasia was found in any of the patients. No relevant side effects or symptoms of estrogenic or androgenic stimulation were recorded. For the climacteric patient needing estrogen therapy, it can be concluded that Org OD 14 is an effective and innocuous medication in the doses used.     

Protection Against Adriamycin-Induced Cardiomyopathy by Carnosine in Rats: Role of Endogenous Antioxidants

The aim of this study was to investigate the effects of carnosine, a biological antioxidant, on the acute cardiac damage induced by a single dose of adriamycin in rats. The experimental design consisted of four groups: Control (saline, i.p.); carnosine (CAR; 10 mg/kg/day, i.p.); adriamycin (ADR; 16 mg/kg on the 14th day, i.p.); carnosine with adriamycin. Carnosine was given 2 weeks before and following adriamycin treatment. Blood samples were collected for analysis of plasma creatine kinase (CK) and plasma antioxidant enzymes, glutathione peroxidase (GSH-Px), Cu, Zn-superoxide dismutase (SOD), and catalase (CAT). The rats were then sacrificed, and the hearts were autopsied for hemodynamic study, ECG, and histopathological examination. Results showed that adriamycin produced evident cardiac damage revealed by hemodynamic change, histological alterations, decreased plasma antioxidant enzymes activities, and increased lipid peroxidation to the control value. Carnosine treatment led to significant attenuation of adriamycin-induced cardiomyopathy revealed by normalization of the LVDP, ST interval, CK, SOD, GSH-Px, CAT, and lipid peroxidation. An increase in oxidative stress and inactivation of SOD, GSH-Px, CAT by a single dose of adriamycin were prevented when carnosine was given 2 weeks before and on the same day adriamycin treatment was administered.     

Comparative evaluation of various analgesics in reducing pain in irreversible pulpitis

It is of interest to evaluate a single dose of three different analgesics compared to placebo in patients with symptomatic irreversible pulpitis. 120 patients were enrolled with severe pain in this prospective clinical trial. Patients were randomly divided into four groups after shaping and cleaning of root canals. This includes placebo, piroxicam 20mg, acetaminophen 325mg with aceclofenac sodium 100mg and acetaminophen 650mg. Participants were given a questionnaire to note the pain scores at various time intervals (6 hrs, 12 hrs, and 24 hrs) along with the respective tablets in a concealed manner. Data thus collected was analyzed for statistical significance. The severity of pain decreased in all the three interventional groups compared to the control group (p <0.01) at 6 hours. Zerodol-P and dolonex showed better pain reduction in comparison to the placebo and dolo 650 group (p <0.05) at 12 and 24 hours. Data shows that both zerodol-P and dolonex groups had similar effects at all time intervals. Thus, a single dose of analgesic such as Zerodol-P and Dolonex following shaping and cleaning of root canals relieved pain at all time intervals of the treatment. However, Dolo 650 performed better during the initial 6hrs after completion of the shaping and cleaning of root canals compared to the placebo.     

神经节苷脂钠对缺血性脑卒中小鼠空间学习记忆能力的影响

摘要 目的：探讨神经节苷脂钠对缺血性脑卒中小鼠空间学习记忆能力的影响。方法：缺血性脑卒中小鼠模型(n=42)随机分为三组-模型组、氟西汀组与神经节苷脂钠组,每组14只小鼠。氟西汀组、神经节苷脂钠组、对照组分别给予10 mg/kg氟西汀与10 mg/kg神经节苷脂钠、等剂量生理盐水腹腔注射,1次/d,持续28 d。结果：氟西汀组、神经节苷脂钠组给药第7 d、14 d、28 d的逃避潜伏期、改良神经损伤严重程度评分(Modified neurological severity score,mNSS)低于模型组(P<0.05),穿越平台次数高于模型组(P<0.05),神经节苷脂钠组与氟西汀组对比差异有统计学意义(P<0.05)。氟西汀组、神经节苷脂钠组给药第28 d的海马组织B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)蛋白相对表达水平高于模型组(P<0.05),BCL2-Associated X(Bax)蛋白相对表达水平、脑卒中相对面积低于模型组(P<0.05),神经节苷脂钠组与氟西汀组对比差异有统计学意义(P<0.05)。结论：神经节苷脂钠在缺血性脑卒中小鼠的应用能促进恢复空间学习记忆能力,缓解神经损伤,抑制海马组织神经元细胞的凋亡,降低脑卒中面积。     

Treatment of nocturnal asthma: the role of sustained-release theophylline and oral beta-2-mimetics

In two double-blind, multiple-dose cross-over studies the therapeutic effects of SR theophylline preparations given once each night (mean 11.2 mg/kg per day) versus twice daily in equal doses (mean 10.3 mg/kg per day) (study I) and SR-terbutaline in equal doses (mean 0.25 mg/kg per day) versus SR theophylline in unequally divided daily doses (mean 5.3 mg/kg morning dose, 10.6 mg/kg evening dose) study II) were compared in 19 patients with nocturnal asthma. At the end of each treatment period drug serum concentrations and PEFR were measured every 2 hr over a 24-hr period. With the twice-daily, equally divided regimen, serum theophylline concentrations were lower at night than during the day (mean 9.4 +/- 0.9 versus 11.3 +/- 1.0 mg/l). With the single evening administration, serum theophylline concentrations were considerably higher at night (Cmax 16.3 +/- 1.4 mg/l) and the circadian variation of PEFR was significantly reduced. PEFR was higher during night and early morning (283 +/- 14 versus 217 +/- 11 l/min, P less than 0.005). During daytime in study II, PEFR values were slightly higher with theophylline than terbutaline. There was no significant difference in peak flow between either treatment during the night and early morning. However, additional use of inhaled beta-2-mimetics because of asthmatic attacks occurred more often during terbutaline (79 times in 8/10 patients) than theophylline treatment (29 times in 5/10 patients). Symptom scores, number of attacks and side-effects clearly favor the theophylline regimen. We conclude that for patients with nocturnal asthma a once-nightly dose of SR theophylline can be sufficient for stabilization of the airways.     

Hydralazine once daily in hypertension.

The effects of hydralazine formulation and dose interval were assessed in 20 patients with hypertension well controlled on conventional hydralazine tablets, 100 mg twice daily, in addition to atenolol and a diuretic. The double-blind study used four regimens crossed over in random order at five-week intervals; placebo; conventional hydralazine 100 mg twice daily; conventional hydralazine 200 mg once daily; and slow-release hydralazine 200 mg once daily. Blood pressure and pulse rate were assessed soon after (2.5 +/- 0.9 h) and immediately before taking hydralazine (previous dose: once daily, 26.5 +/- 0.9 h; twice daily, 13.6 +/- 2.0 h). Seventeen patients completed the study. All hydralazine regimens were associated with significant falls in blood pressure. Once-daily treatment with conventional hydralazine was unsatisfactory, as its hypotensive effect waned at 24 h; there was a significant difference between the peak and trough effects on blood pressure and pulse in rapid acetylators. Compared with placebo twice-daily conventional hydralazine and once-daily slow-release hydralazine gave satisfactory control for 24 hours in both rapid and slow acetylators, though the hypotensive effect was larger in the slow acetylators. It is concluded that there is no need to administer hydralazine more than twice daily.     

Physical anomalies and developmental delays in nonhuman primate infants exposed to weekly doses of ethanol during gestation

Ethanol was orally administered once per week to 54 gravid pigtailed macaques (Macaca nemestrina) in doses of 0.0, 0.3, 0.6, 1.2, 1.8, 2.5 or 4.1 gm/kg from the 1st week in gestation or in doses of 2.5, 3.3, or 4.1 gm/kg from the 5th week. Mean maternal mean peak plasma ethanol concentrations (MPPEC) ranged from 24 +/- 6 mg/dl at the 0.3 gm/kg dose to 549 +/- 71 mg/dl at the 4.1 gm/kg dose. Thirty-three viable infants were followed from birth to 6 months of age and assessed for growth, health, congenital anomalies and developmental rate. Facial anomalies, growth deficiency, or central nervous system dysfunction were found in 57% of the alcohol-exposed animals. No animal showed all the features of the human fetal alcohol syndrome. Ten of the twelve animals (83%) with mean MPPEC above 140 mg/dl had evidence of a teratogenic impact. The animals with full gestational exposure to ethanol and mean MPPEC between 140 and 249 mg/dl had much more severe and consistent cognitive abnormalities than the animals with delayed gestational exposures, even though the latter were exposed to mean MPPEC between 260 and 540 mg/dl. Conclusions from this study included: 1) ethanol-related behavioral teratogenesis occurred without accompanying physical anomalies, 2) measurable teratogenic effects from weekly exposures occurred only at intoxicating doses of ethanol, and 3) early gestational exposure to ethanol appeared to be more damaging to cognitive function than later and considerably greater alcohol exposure.     

Estradiol treatment and its interaction with the cholinergic system: effects on cognitive function in healthy young women

The steroid hormone estradiol has been shown to modulate cognitive function in both animals and humans, and although the exact mechanisms associated with these effects are unknown, interactions with the cholinergic system have been proposed. We examined the neurocognitive effects of short-term estradiol treatment and its interaction with the cholinergic system using the muscarinic receptor antagonist scopolamine in healthy young women. Thirty-four participants (Mean age ± SD = 22.4 ± 4.4) completed baseline cognitive assessment and then received either 100 μg/day transdermal estradiol or transdermal placebo for 31 days. On days 28 and 31 of treatment, further cognitive assessment was performed pre- and 90 min post-scopolamine (0.4 mg) or placebo (saline) injection, under a randomized double-blind placebo-controlled design. Short-term estradiol treatment significantly enhanced spatial working memory with a trend for improvement in long-term verbal learning and memory. Overall, estradiol treatment did not protect against or attenuate the scopolamine-induced impairments in the cognitive domains assessed. Findings suggest that estrogen has minimal effects on cholinergic-mediated cognitive processes following short-term treatment. Effects of estradiol treatment may be dependent on age, dose of estradiol, integrity of cholinergic innervation and baseline endogenous estrogen levels, which may in part explain the inconsistent findings in the literature.