Adrenocorticotropic Hormone Group Research Articles

Neurobehavioral and molecular changes in a rodent model of ACTH-induced HPA axis dysfunction

Hypothalamic-pituitary-adrenal (HPA) axis dysregulation is linked to the pathophysiology of depression. Although exogenous adrenocorticotropic hormone (ACTH) is associated with a depressive-like phenotype in rodents, comprehensive neurobehavioral and mechanistic evidence to support these findings are limited. Sprague-Dawley rats (male, n = 30; female, n = 10) were randomly assigned to the control (male, n = 10) or ACTH (male, n = 20; female n = 10) groups that received saline (0.1 ml, sc.) or ACTH (100 μg/day, sc.), respectively, for two weeks. Thereafter, rats in the ACTH group were subdivided to receive ACTH plus saline (ACTH_S; male, n = 10; female, n = 5; 0.2 ml, ip.) or ACTH plus imipramine (ACTH_I; male, n = 10; female, n = 5;10 mg/kg, ip.) for a further four weeks. Neurobehavioral changes were assessed using the forced swim test (FST), the sucrose preference test (SPT), and the open field test (OFT). Following termination, the brain regional mRNA expression of BDNF and CREB was determined using RT-PCR. After two-weeks, ACTH administration significantly increased immobility in the FST (p = 0.03), decreased interaction with the center of the OFT (p < 0.01), and increased sucrose consumption (p = 0.03) in male, but not female rats. ACTH administration significantly increased the expression of BDNF in the hippocampus and CREB in all brain regions in males (p < 0.05), but not in female rats. Imipramine treatment did not ameliorate these ACTH-induced neurobehavioral or molecular changes. In conclusion, ACTH administration resulted in a sex-specific onset of depressive-like symptoms and changes in brain regional expression of neurotrophic factors. These results suggest sex-specific mechanisms underlying the development of depressive-like behavior in a model of ACTH-induced HPA axis dysregulation.

Comparison of Different Doses of ACTH Used in ACTH Stimulation Test to Determine the Subtypes of Primary Aldosteronism

To compare the diagnostic value of adrenocorticotropic hormone (ACTH) stimulation test (AST) with different doses of ACTH combined with midnight administration of 1 mg dexamethasone for the determination of the subtypes of primary hyperaldosteronism (PA). This is a prospective observational study. Patients diagnosed with PA in the Department of Endocrinology, the First Medical Center of of Chinese PLA General Hospital from January 1, 2020 to September 30, 2022 underwent AST with different doses of ACTH. All patients received 1 mg dexamethasone at midnight for inhibition. Then, the patients were randomly assigned to 25-unit and 50-unit ACTH treatment groups by a ratio of 1:2. Subtype classification and diagnosis of aldosterone-producing adenoma (APA) and idiopathic hyperaldosteronism (IHA) was made on the basis of adrenal venous blood samples and/or postoperative pathology and clinical follow-up findings. Receiver operating characteristics (ROC) curves were plotted to examine the diagnostic efficacy and the difference of AST by varying doses of ACTH in distinguishing APA and IHA. A total of 82 patients, including 49 patients with APA (59.8%) and 33 patients with IHA (40.2%), were enrolled. There were 29 patients in the 25-unit ACTH group (35.4%) and 53 patients in the 50-unit ACTH group (64.6%). There were no significant differences in age, sex, blood pressure, minimum serum potassium, and biochemical parameters between the 25-unit and 50-unit groups. After ACTH stimulation, plasma aldosterone concentration (PAC), cortisol (F), and PAC/F at different points of time showed no statistical difference between the two groups (P>0.05). The area under the curve (AUC) of PAC in the 25-unit group was higher than that of PAC/F. The AUC of PAC reached the maximum at 90 minutes (0.948, 95% confidence interval [CI]: 0870-1.000) and the optimal cutoff was 38.0 ng/dL, which had a sensitivity of 92.9% and a specificity of 86.7% for differentiating APA and IHA. Similar to the 25-unit group, the maximum AUC of PAC in the 50-unit group was greater than that of PAC/F. The AUC of PAC reached the maximum 90 minutes (0.930, 95% CI: 0.840-0.994) and the optimal cutoff was 39.6 ng/dL, which had a sensitivity of 91.2% and a specificity of 83.3%. The AUC of PAC at different points of time in the 25-unit ACTH group (0.862-0.948) was greater than that of 50-unit ACTH group (0.823-0.930), but the difference was not statistical significance. AST with 25-unit or 50-unit ACTH combined with small-dose dexamethasone can be used in PA subtype determination, ie, differentiation between APA and IHA. The optimal PAC cut-off values for 25-unit or 50-unit ACTH are similar, being 38.0 ng/dL and 39.6 ng/dL, respectively, and both cutoff values show higher sensitivity and specificity at 90 min. The AST with 25-unit ACTH has the smaller dose and the better safety. Therefore, it is recommended for the diagnosis of PA subtypes.

Endocrine characteristics and risk factors of type 2 diabetes complicated with gastrointestinal autonomic neuropathy: A single-center retrospective study.

To analyze endocrine characteristics and risk factors of type 2 diabetes mellitus (T2DM) gastrointestinal autonomic neuropathy. A total of 202 patients with T2DM with obesity who were hospitalized at our institute between January 2019 and June 2021 were selected. Based on adrenocorticotropic hormone (ACTH) levels, 102 patients were placed in the ACTH abnormal group and 100 patients were placed in the non-ACTH abnormal group. Ninety-five healthy adults without diabetes, hyperlipidemia, osteoporosis, or endocrine system abnormalities who were examined at our hospital during the same period were selected as the control group. Fasting plasma glucose (FPG), fasting insulin (FINS), triglycerides (TG), total cholesterol (TC), homeostasis model assessment of insulin resistance (HOMA-IR), ACTH level, body mass index (BMI), and bone mineral density (BMD) were measured to evaluate endocrine characteristics and risk factors. BMI, FPG, FINS, HOMA-IR, TG, TC, and ACTH levels in the abnormal ACTH group were significantly higher than those in the other 2 groups, while BMD was significantly lower than that in the other 2 groups (all P < .05). BMI, FPG, FINS, HOMA-IR, TG, TC, and ACTH in the non-ACTH abnormal group were significantly higher than those in the control group, whereas BMD was significantly lower than that in the control group (all P < .05). The plasma ACTH level in patients with abnormal ACTH levels was significantly positively correlated with BMI, FPG, FINS, HOMA-IR, TG, and TC and negatively correlated with BMD (all P < .05). Multivariate regression analysis showed that BMI, advanced age, FINS, TG, and FPG were risk factors for ACTH abnormalities in patients with diabetes (odds ratio > 1, all P < .05). BMI, advanced age, FINS, TG and FPG are the risk factors of abnormal ACTH in T2DM patients with gastrointestinal autonomic neuropathy.

Real-word adrenocorticotropic hormone treatment for childhood-onset nephrotic syndrome.

Current first-line anti-proteinuric treatments do not produce a satisfactory therapeutic effect in a considerable number of patients with nephrotic syndrome (NS). Interest in adrenocorticotropic hormone (ACTH) for the treatment of NS has recently been revived. The present study investigated the efficacy and safety of ACTH treatment in children with frequent relapsing NS (FRNS), steroid-dependent NS (SDNS), and steroid-resistant NS (SRNS). The ACTH treatment group was comprised of NS patients receiving ACTH treatment. Patients with serum cortisol concentrations <85.3 nmol/L and who had not received ACTH treatment previously were enrolled in the control group from January 2018 to January 2021. The maintenance dose of prednisone, the number of disease recurrences, the time of first disease relapse, immunosuppressant use, serum cortisol levels, and adverse events were recorded in both groups. Fifty-one patients were included in the ACTH group, and twenty-one patients were enrolled in the control group. Concurrent treatment with one or more immunosuppressive and/or cytotoxic treatments occurred in 92.2% and 85.7% of patients in the ACTH and control groups, respectively, throughout the study period. A greater reduction in the prednisone maintenance dose was observed in the ACTH group compared with the control group after 1 year of follow up (0.603 ± 0.445 mg/kg vs. 0.267 ± 0.500 mg/kg, p = 0.006). During the one-year study period, fewer participants experienced one or more disease relapses in the ACTH group (45.1%) compared to the control group (76.2%, odds ratio = 3.896, p = 0.016). The number of disease recurrences per patient in the ACTH group was less than that in the control group (median difference = -1, p = 0.006). The mean length of remission was 8.902 m and 7.905 m in the ACTH group and control group, respectively. A log-rank test showed a longer relapse free survival for patients in the ACTH group (p = 0.046), but the Breslow test showed no significant difference between groups (p = 0.104). Ten patients in the ACTH group successfully discontinued all drug therapies. No patients in the control group were able to discontinue drug therapy as of February 2022. ACTH, combined with multiple drugs, is effective at reducing the prednisone maintenance dose and may effectively prevent disease relapses in childhood NS.

A Randomized Controlled Trial on the Study of Effectiveness and Safety of Hormonal (ACTH) Treatment Alone versus Hormonal (ACTH) with Levetiracetam for Epileptic Spasms.

Background West's syndrome (WS) is a triad of epileptic spasms (ESs), psychomotor delay, and hypsarrhythmia. The treatment of ESs is still controversial. Hence, we designed a randomized controlled trial (RCT) to compare the outcomes in children with WS treated with adrenocorticotropic hormone (ACTH) alone versus ACTH and levetiracetam (LEV). Objectives To compare the treatment outcomes and side effects in children treated with ACTH alone versus ACTH and LEV. Methods This prospective randomized controlled trial was conducted from December 2017 to May 2019 in tertiary care center, Bangaluru. Children from 2 months to 5 years of age, diagnosed with WS were included. Fifty children in each group were analyzed for efficacy and side effects. Results There was no difference in the baseline characteristics in both groups. There was no difference in spasms response at the end of 2 weeks between the groups (88 vs. 82%) with p -value of 0.813. The relapse rates were less in ACTH and LEV group (20%) compared with ACTH alone (22%) but statistically not significant ( p > 0.1). There was no difference observed in subsequent epilepsy rates (18%) in ACTH versus 19% in ACTH with LEV group ( p > 0.1) and side effects. There was improvement in milestones 48% in ACTH with LEV group versus 37% in ACTH alone however statistically not significant ( p > 0.1). Conclusion There was no difference in children treated with ACTH alone versus ACTH and LEV in terms of control of spasms and subsequent epilepsy rates. The relapse rate is less, and developmental outcome is better in ACTH with LEV group but statistically not significant.

Efficacy of adrenocorticotropic hormone in children with frequently relapsing or steroid-dependent nephrotic syndrome

Objective: To investigate the efficacy and safety of adrenocorticotropic hormone (ACTH) in children with frequently relapsing or steroid-dependent nephrotic syndrome. Methods: The clinical data of 38 children with frequently relapsing or steroid-dependent nephrotic syndrome who were admitted to the Department of Nephrology, the Children Hospital, Zhejiang University School of Medicine from January 2015 to December 2020 were retrospectively analyzed. The general information, clinical manifestations, laboratory data of the children and follow-up (till 12 months after treatment) were collected. The patients were divided into ACTH group and Glucocorticoid (GC) group according to treatment plan. Cumulative remission, average recurrence rate, GC dosage, height and weight change and peripheral blood CD19+B lymphocyte count were compared between the two groups to evaluate the efficacy and adverse reactions of ACTH. Fisher's exact test, t test or rank sum test was used for comparison between groups. Results: Among the 38 patients, 28 were male and 10 were female, aged 84 (24, 180) months; 19 were in ACTH group and 19 were in GC group. The cumulative remission rate of 12 months in ACTH group was higher than that in GC group (9/19 vs. 2/19,χ²=6.81,P=0.009), the average recurrence rate was lower than that in GC group ((0.7±0.8) vs. (1.7±1.1) times, t=-3.27, P=0.011), and the average dosage of GC was lower than that in GC group ((0.27±0.16) vs. (0.51±0.27) mg/(kg·d), t=-3.21, P=0.014). The increase in height was higher than that in the GC group (4 (3,5) vs. 3 (2, 3) cm/year, Z=2.58, P=0.010), and the peripheral blood CD19+B lymphocyte count was lower than that in the GC group ((223±149)×106 vs. (410±213)×106/L,t=-3.35, P=0.009). In safety, 19 cases had transient decreased urine volume, 7 cases had hyperglycemia, and 3 cases had hypertension during the infusion of ACTH, which could be relieved after drug withdrawal. Conclusion: ACTH has a better effect on children with frequently relapsing or steroid-dependent nephrotic syndrome, which can improve cumulative sustained remission rate, lower relapses rate and decrease the dosage of GC, with good safety.

Differences in thyroid function tests after administration of a single supraphysiological dose of ACTH or a high dose of a synthetic steroid

ObjectiveAlthough the suppressive action of synthetic steroids on the hypothalamus-pituitary-thyroid (HPT) axis is established, little is known regarding the effect of the administration of synthetic adrenocorticotropic hormone (ACTH). DesignIn the context of a randomized, open label, comparative study assessing the efficacy and safety of ACTH and betamethasone in the treatment of hospitalized patients with acute gout, we compared the effects of these agents on thyroid function tests. MethodsSerum TSH, total T4 and T3 and cortisol were measured before and at 24 and 48 h after a single intramuscular dose of synthetic ACTH (1 mg) or betamethasone (6 mg), in 38 hospitalized patients with acute gout and normal thyroid function. ResultsThe final analysis included 32 patients, due to missing data. The ACTH and betamethasone groups did not differ regarding the mean age, gender, severity of gout attack, and baseline thyroid parameters. In the ACTH group TSH and T4 were significantly decreased at 24 and at 48 h compared to baseline, while T3 was decreased at 24 but not at 48 h. In the betamethasone group T3 remained stable; TSH and T4 decreased significantly from baseline levels at 24 h; at 48 h, TSH had returned to and T4 showed a partial rebound towards pre-treatment values. ConclusionsA single IM administration of 1 mg of synthetic ACTH has more profound and prolonged effects on the HPT axis, lasting for at least 48 h, compared to a single IM dose of 6 mg betamethasone.

Mass spectrometry‐based cortisol profiling during adrenal venous sampling reveals misdiagnosis for subtyping primary aldosteronism

Primary aldosteronism (PA) is a common form of secondary hypertension. Adrenal venous sampling (AVS) is the gold standard for subtyping PA. This study aimed to determine whether there is a difference between immunoassays and liquid chromatography-mass spectrometry (LC-MS/MS) methods for measuring cortisol levels that affect the judgement of AVS. This was a retrospective study. Included 72 patients who were diagnosed with PA and had undergone AVS. Patients were grouped according to whether they received adrenocorticotropic hormone (ACTH) stimulation during AVS, and the cortisol results were measured using immunoassay and LC-MS/MS. There were 48 patients in the without ACTH stimulation group and 24 in the post-ACTH stimulation group during AVS (bilateral adrenal vein cannulation success rate, 56.25% vs. 83.33%). ACTH stimulation was beneficial for increasing the success rate of AVS (p < .001). Immunoassays were linearly correlated with LC-MS/MS when cortisol concentrations were <1750 nmol/L (r = .959, p < .001). When cortisol concentrations were >17,500 nmol/L, no correlation was found between the two methods (p = .093). The two methods were consistent for the detection of cortisol for evaluating the success of cannulation for AVS. Five percent of patients showed discordant lateralization of aldosterone production according to the cortisol LC-MS/MS and immunoassay results in the without ACTH group, and 15% showed discordant lateralization in the post-ACTH group. The immunoassay method can be used to determine whether cannulation is successful. The final decision for lateralization may be more appropriate based on LC-MS/MS results.

The value of serum dehydroepiandrosterone sulfate in differential diagnosis of Cushing's syndrome

Objective: To evaluate the changes and diagnostic value of serum dehydroepiandrosterone sulfate (DHEAS) in Cushing's syndrome (CS) with different etiologies. Methods: The study retrospectively recruited patients diagnosed as CS in Drum Tower Hospital affiliated to Nanjing University Medical School between January 2012 and June 2019, including 36 patients (8 males, 28 females, with an average age of 44 years) with Cushing disease (CD) and 64 patients (6 males, 58 females, with an average age of 39 years) with adrenal CS (ACS). Meanwhile, 97 patients diagnosed as nonfunctional adrenal adenoma (NFA) were also included as controls. Clinical characteristics, laboratory data, adrenocorticotropic hormone (ACTH), serum DHEAS level and sex-and age-adjusted DHEAS ratio of the three groups were collected. The sensitivity and specificity of DHEAS and its ratio in differential etiology diagnosis of CS were compared using receiver operating characteristic (ROC) curve analysis. Results: Compared to NFA group, ACS patients had lower DHEAS levels [0.39 (0.39, 0.63) μmol/L vs 2.96 (1.92, 4.60) μmol/L, P<0.01] and lower DHEAS ratio [0.58 (0.27, 0.98) vs 3.95 (3.08, 6.83), P<0.01]. DHEAS [6.49 (4.32, 11.63) μmol/L] and DHEAS ratio [9.17 (4.49, 15.41)] in CD patients were significantly higher compared to those in NFA and ACS patients (all P<0.01). There were 53 ACS patients (82.8%) with suppressed ACTH level (<2.2 pmol/L) and 11 patients (17.2%) with normal/high ACTH level (≥2.2 pmol/L). The level of 24 hour urine free cortisol in normal/high ACTH level group was lower than the suppressed ACTH group [(1 299±511) nmol/24 h vs (1 972±876) nmol/24 h, P=0.04]. No significant differences were found in the DHEAS and DHEAS ratio between the two groups. ROC analysis showed that the area under the curve of serum DHEAS and DHEAS ratio in diagnosing ACS from CD was 0.997 and 0.990, respectively. The optimal cut-off values for DHEAS and its ratio were 2.06 μmol/L and 2.10, respectively. The diagnostic sensitivity and specificity of DHEAS were 97.5% and 100%, and those of DHEAS ratio were 95.0% and 100%, respectively. Conclusion: There are significant differences in serum DHEAS level and DHEAS ratio between ACS and CD patients, which might be used as indicators for the identification of the two main CS etiologies, especially in the identification of ACS patients without plasma ACTH suppression from CD patients.

Comparison of the Effect of High and Low Doses of Adrenocorticotropic Hormone (ACTH) in the Management of Infantile Spasms.

Infantile spasms can have irrecoverable adverse effects on a child's brain. Adrenocorticotropic hormone (ACTH) is the most common first-line medication for the treatment of infantile spasms. However, the suitable dose and duration of treatment continue to be debated among specialists. Since high doses of ACTH, which are commonly used, can produce more side effects, lower doses are preferred. The aim of this study was to determine the effect and extent of complications caused by high and low doses of ACTH in children with infantile spasms. This clinical trial was performed on 32 infants with infantile spasms, aged 1.5-18 months. The subjects were divided into high- and low-dose ACTH groups. Treatment continued for two months. The therapeutic effects and complications were then compared over 18 months. The results indicated no significant difference between the groups in terms of the short-term prognosis of convulsions, final prognosis of patients with spasm relapse, EEG changes after treatment, and post-treatment development of hypertension. On the other hand, there was a significant difference in the frequency distribution of restlessness intensity and becoming Cushingoid, which were more frequent in the high-dose group. The results indicated that high- and low-dose ACTH are equally effective in controlling spasms, yet the low dose causes fewer side effects.

Incidence of Hypertension Among Children Treated With Adrenocorticotropic Hormone (ACTH) or Prednisolone for Infantile Spasms.

Children with infantile spasms are often treated with hormonal therapies including adrenocorticotropic hormone (ACTH) and prednisolone. These have numerous systemic side effects including hypertension and, rarely, fatal cardiomyopathy; however, the incidence of these side effects has not been well described. This study aims to quantify the incidence and short-term sequelae of hypertension in this population. A retrospective chart review was performed at a single institution. Children 2 months to 2 years old with newly diagnosed infantile spasms treated from 2013 to 2017 were included. Variables collected included age, sex, etiology and treatment of infantile spasms, documented or missed diagnosis of hypertension, treatment of hypertension, echocardiogram results, referrals for hypertension, and persistence of hypertension 2 to 4 months after treatment. Analyses included descriptive statistics with percentiles, means, and medians. Differences between groups were assessed using Fisher exact tests. Hypertension occurred in 34/77 children (44%) during treatment with ACTH and 4/11 children (36%) during treatment with prednisolone. No child developed hypertension during treatment with nonhormonal therapies. The incidence of hypertension between ACTH and prednisolone groups was not significantly different (P = .75). The incidence of hypertension was significantly higher in the ACTH and prednisolone groups compared to the nonhormonal group (P < .001 for each). Sixteen children received echocardiograms, with no cases of cardiomyopathy. Two children had persistent hypertension at 2 months after discontinuation of hormonal therapy. Hypertension is a very common side effect of hormonal therapy for infantile spasms; however, few developed long-term hypertension and none developed cardiomyopathy. Further study is needed to determine the role of antihypertensive treatment for hormone-related hypertension.

Effect of repeated adrenocorticotropic hormone administration on reproductive function and hair cortisol concentration during the estrous cycle in goats

Measurement of the cortisol concentration in hair has been used as an index of chronic stress in several species including humans, wildlife and domestic animals. However, how accurately the cortisol concentration in hair reflects the changes in circulating cortisol concentrations has not been well documented. The objective of the present study was to examine the effect of repeated adrenocorticotropic hormone (ACTH) administration on the reproductive function during the estrous cycle and hair cortisol concentrations in goats. In experiment 1, goats were administered ACTH (0.625 IU/10 kg of body weight, n = 6) or saline (n = 6) intramuscularly once a day for 7 days on Day 11–17 of the estrous cycle (day 0 was the day of ovulation). In experiment 2, goats were administered ACTH (0.625 IU/10 kg of body weight, n = 6) or saline (n = 6) intramuscularly twice a day on Day 11–24 of the estrous cycle. Blood samples were collected 0, 0.5, and 6 h after first administration to determine the circulating cortisol concentrations. Hair was clipped at 0, 1, and 2 months after the start of administration. In both experiments, the plasma cortisol concentration increased at 0.5 h and returned to baseline at 6 h after ACTH administration. During the experiments, estrus was observed in most animals in ACTH and saline groups (6/6 and 4/6 in experiment 1 and 5/6 and 6/6 in experiment 2, respectively) and ovulation was observed in all goats examined. However, the number of ovulatory follicles was significantly different between the ACTH and saline groups, and the maximal diameter of ovulatory follicles tended to be different (P = .07) between the ACTH and saline groups. In experiment 1, the hair cortisol concentration was not influenced by the ACTH administration throughout the sampling period. In experiment 2, the hair cortisol concentration in the ACTH group was greater at 1 month after administration than the pre-administration value, but was not significantly different at 2 months. These results suggest that repeated ACTH administration affects the development and ovulatory process of ovarian follicles and analysis of the hair cortisol concentration can be used for assessing relatively long-term changes in cortisol concentration in the circulation.

Adrenocorticotropic hormone versus methylprednisolone added to interferon β in patients with multiple sclerosis experiencing breakthrough disease: a randomized, rater-blinded trial.

The objective of this study was to evaluate monthly intramuscular adrenocorticotropic hormone (ACTH) gel versus intravenous methylprednisolone (IVMP) add-on therapy to interferon β for breakthrough disease in patients with relapsing forms of multiple sclerosis. This was a prospective, open-label, examiner-blinded, 15-month pilot study evaluating patients with Expanded Disability Status Scale (EDSS) score 3.0-6.5 and at least one clinical relapse or new T2 or gadolinium-enhanced lesion in the previous year. Twenty-three patients were randomized to ACTH (n = 12) or IVMP (n = 11) and completed the study. The primary outcome measure was the cumulative number of relapses. Secondary outcomes included EDSS, Mental Health Inventory (MHI), plasma cytokines, MS Functional Composite (MSFC), Quality-of-Life (MS-QOL) score, bone mineral density (BMD), and new or worsened psychiatric symptoms per month. Brain magnetic resonance imaging was analyzed post hoc. This was a preliminary and small-scale study. Relapse rates differed significantly [ACTH 0.08, 95% confidence interval (CI) 0.01-0.54 versus IVMP 0.80, 95% CI 0.36-1.75; rate ratio, IVMP versus ACTH: 9.56, 95% CI 1.23-74.6; p = 0.03]. ACTH improved (p = 0.03) MHI (slope 0.95 ± 0.38 points/month; p = 0.02 versus slope -0.38 ± 0.43 points/month; p = 0.39). On-study decreases (all p < 0.05) in eight cytokine levels occurred only in the ACTH group. However, on-study EDSS, MSFC, MS-QOL, BMD, and MRI lesion changes were not significant between groups. Psychiatric symptoms per patient were greater with IVMP than ACTH (0.55, 95% CI 0.12-2.6 versus 0; p < 0.0001). Other common adverse events were insomnia and urinary tract infections (IVMP, seven events each) and fatigue or flu symptoms (ACTH, five events each). This study provided class II evidence that ACTH produced better examiner-assessed cumulative rates of relapses per patient than IVMP in the adjunctive treatment of breakthrough disease in multiple sclerosis.

Efficacy of high-dose ACTH versus low-dose ACTH in infantile spasms: A meta-analysis with direct and indirect comparison of randomized trials

To determine the influence of dose on the efficacy and safety of adrenocorticotropic hormone (ACTH) for infantile spasms. We searched six relative databases. Randomized controlled trials and crossover studies, which meet our inclusion criteria were considered for inclusion. The quality of included trials was evaluated with respect to sequence generation, allocation concealment, blinding and incomplete data. We performed both direct and indirect meta-analysis to evaluate the efficacy and safety of high-dose and low-dose of ACTH. Results: A total of 168 reports were retrieved, and after scrutiny seven reports of trials containing three head to head comparisons, two vigabatrin comparisons and two prednisone comparisons were included. In direct comparison meta-analysis, studies were homogenous (P >; 0.10). There was a trend towards more complete cessation of spasms in the low-dose ACTH treated group (RR: 0.86, 95% confidence interval [CI]: 0.57–1.31), though statistically not significant. Relapse was more frequent in low-dose ACTH groups (RR: 0.69, 95% CI: 0.22–2.19) but without demonstrating statistic difference. No difference was present for disappearance of hypsarrhythmia between high and low-dose ACTH (RR: 0.93, 95% CI: 0.57–1.54). In indirect meta-analysis the comparison of complete cessation of spasms between high-dose and low-dose ACTH from indirect meta failed to show any significant difference. There was no significant difference between two dosages with respect to relapse rates. High-dose ACTH produced more adverse effects than low-dose group. A low-dose ACTH therapy might be preferable for its comparative efficacy and lower risk of adverse effects.

Blood plasma collected after adrenocorticotropic hormone administration during the preovulatory period in the sow negatively affects in vitro fertilization by disturbing spermatozoa function

Successful fertilization is essential for reproduction and might be negatively affected by stressful events, which could alter the environment where fertilization occurs. The aim of the study was to determine whether an altered hormonal profile in blood plasma caused by adrenocorticotropic hormone (ACTH) administration could affect in vitro fertilization in the pig model. In experiment 1, gametes were exposed for 24 hours to plasma from ACTH-treated, non-ACTH–treated sows, or medium with BSA. Fertilization, cleavage, and blastocyst rates were lower in the ACTH group compared with the no ACTH or BSA control groups (P < 0.01). In experiment 2, the exposure of matured oocytes for 1 hour before fertilization to the same treatments did not have an impact on their ability to undergo fertilization or on embryo development. In experiment 3, spermatozoa were incubated for 0, 1, 4, and 24 hours under the same conditions. There was no effect of treatment on sperm viability. The percentage of acrosome-reacted spermatozoa remained higher in the ACTH group compared with the non-ACTH–treated group through the incubation period (P < 0.001). Protein tyrosine phosphorylation (PTP) patterns were also affected by treatment (P < 0.001). The presence of an atypical PTP pattern was higher in the ACTH group at all the analyzed time points compared with the BSA and no ACTH groups (P < 0.001). In conclusion, this altered environment may not affect oocyte competence but might affect the sperm fertilizing ability through alterations in the acrosome reaction and correct sequence of PTP patterns.

VALIDATION OF A FECAL GLUCOCORTICOID METABOLITE ASSAY FOR COLLARED PECCARIES (PECARI TAJACU)

The possibility of assessing endogenous adrenal activity in the collared peccary (Pecari tajacu) was tested by using an adrenocorticotropic hormone (ACTH) challenge in a fecal glucocorticoid metabolite (FGM) assay. Feces were collected from 12 captive adult male peccaries beginning 48 hr prior to challenge; six of these animals received the challenge as an ACTH injection and the other six were injected with saline solution. Feces collection ended 120 hr after injections. As a control, feces were collected for eight consecutive days from another six adult male peccaries that remained in their original mixed-sex herds in semiconfined paddocks. All feces samples were freeze-dried, extracted by an ethanol vortex method, and assayed for glucocorticoids by means of an enzyme immunoassay. FGM concentrations were compared between the treatments by a repeated measures analysis of variance (ANOVA) followed by a post hoc Tukey test. The assay is reliable but, instead of the usual proportion of 1:50 in ethanol (fecal mass:solvent), 1:10 is recommended for best extraction of FGM. Baseline FGM concentrations were similar among the ACTH, saline, and control treatments (29.7 +/- 11.2 ng/g(-1) dry feces) during the 48 hr before the challenge. The ACTH group reached an FGM excretion peak at 24 hr post-treatment, followed by a decline, while in the control and saline groups FGM levels remained relatively constant. Therefore, the fecal glucocorticoid metabolite assay reflects endogenous adrenal activity in the collared peccary and is a powerful tool for noninvasive stress monitoring in peccaries.

Monthly Pulse Adrenocorticotropic Hormone (ACTH) or Methylprednisolone Therapy for Long-Term Treatment of Multiple Sclerosis as an Add-On Therapy to beta-Interferons: Current Status of the Pilot Study (P04.147)

Objective: This pilot study is designed to evaluate the efficacy and safety of monthly adrenocorticotropic hormone (ACTH) treatment added to beta-interferon in breakthrough multiple sclerosis (MS) compared with monthly methylprednisolone (MP), the most commonly used therapy. Background ACTH gel is approved for treatment of MS relapses. There is mounting evidence that ACTH may have a variety of immune-modulating mechanisms relevant to the MS disease course. However, no clinical studies have evaluated ACTH pulse therapy for MS. Design/Methods: This single-center, examiner-blinded study enrolled MS patients receiving ongoing beta-interferon treatment with Expanded Disability Status Scale (EDSS) scores of 3.0-6.5 and ≥1 relapse or new T2 or Gadolinium-enhanced lesion within the previous year. Patients were randomly assigned to open-label ACTH (80 units intramuscularly once/day x 3 consecutive days) or MP (1 gram IV x 1 dose) monthly for 12 months, with assessments every 3 months. Outcomes include relapse rate (primary); EDSS, MS Functional Composite, and MS Quality of Life scores; MRI; bone density; and side effects. We report the current status of the study. Results: Of 26 patients enrolled (ACTH: n=14, mean±SD EDSS 4.4±1.4; MP: n=12, mean EDSS 4.7±1.4), 18 have ≥6 months of data (ACTH: n=10, MP: n=8). During 6 months of pulse therapy, there were 0 relapses in the ACTH group and 4 relapses (in 2 patients) in the MP group. Mean EDSS score at month 3 was 3.9±2.0 (p=0.028; n=12) for ACTH and 3.9±1.6 (p=0.058; n=10) for MP. EDSS at month 6 was 4.4±1.8 (p=0.075) for ACTH and 4.2±2.0 (p=0.401) for MP. Seventeen side effects were reported from 9 patients in the ACTH group and 13 side effects were reported by 6 patients in the MP group. Conclusions: These data suggest a potential role for ACTH pulse therapy in treating breakthrough MS. Supported by: Questcor Pharmaceuticals, Inc. Disclosure: Dr. Berkovich has received personal compensation for activities with Acorda Therapeutics, Bayer Pharmaceuticals Corporation, Biogen Idec, Teva Neuroscience and Questcor Inc. as a speaker. Dr. Berkovich has received research support from Questcor Inc. Dr. Amezcua has received personal compensation for activities with Bayer Pharmaceuticals Corporation, Biogen Idec, Teva Neuroscience, and Serono/Pfizer Inc. Dr. Fernandez has nothing to disclose. Dr. Subhani has nothing to disclose. Dr. Kravtsova has nothing to disclose.

Immunoglobulin changes in boars exposed to administration of levamisole and exogenous adrenocorticotropic hormone

The objective of this paper is to determine the effect of levamisole (LEV) immunoglobulin concentration in the serum of boars exposed to three-day stress induced by the adrenocorticotropic hormone (ACTH). Boars were assigned to four groups (n=7). The first group received LEV for three days (2.5 mg/kg BW), the second group received ACTH (10 g/kg BW) for three days and the third group received LEV for three consecutive days and ACTH on the three following days (2.5 mg/kg BW ; ACTH 10 g/kg BW). The control group received saline solution (1 mL) during the experiment (six days). Concentrations of total protein, globulin, albumin and immunoglobulin concentration (IgG, IgA and IgM) were determined during levamisole, ACTH and levamisole + ACTH combined treatment administration, and during the 16-day post- administration period. Total protein levels were significantly increased during the stress period and over the next sixteen days (p0.01 to p0.0001, respectively) in the ACTH group of boars. However, total protein levels were significantly elevated in the LEV+ACTH group on day 1 and 2 of ACTH injection (p0.05 and p0.001) and on day 5 and the last day of measurement (p0.001 and p0.01). However, the three-day administration of levamisole (LEV) stimulated a statistically significant increase of protein concentrations compared to control values after levamisole treatment, on days 2 and on days 11, 15 and 24 (p0.01, all). Levels of serum albumin were not affected with levamisole or ACTH treatment. According to determined total protein concentrations, globulin concentrations were significantly increased throughout and after the ACTH treatment in the second group (p0.01 to p0.0001, respectively). Also, globulin concentrations were significantly elevated in the LEV+ACTH group throughout the three-day ACTH treatment and in the 16-day post-administration period (p0.05 to p0.01, respectively). The three-day levamisole administration resulted in an increase of globuline concentrations (LEV). However, there were no statistically significant differences in comparison to control pigs. ACTH treatment significantly elevated serum IgG concentration during the stress period (p0.001 to p0.0001, respectively) and over the next 16 days (p0.01 to p0.0001, respectively). However, in the LEV+ACTH group of boars, IgG levels were significantly elevated on days 1 and 3 of ACTH injection (p0.05 and p0.01) and days 1 and 5 in the post-treatment period (p0.01 and p0.05). Administration of levamisole (LEV) had no significant impact on IgG levels compared to the control boar group. However, a statistically significant increase in IgA concentrations in boars treated with levamisole (LEV) was determined on day 2 (p0.02) and day 11 (p0.05) after administration of levamisole compared to animals of the LEV+ACTH group. Furthermore, boars receiving levamisole before stress induction (LEV+ACTH) on day 5 after stress induction had IgM concentrations statistically significantly higher (p0.05) than animals treated only with ACTH. These results show that levamisole may stimulate the immune system in swine and immunoglobulin in the serum of boars exposed to stress.

208 IN VITRO FERTILIZATION UNDER SIMULATED STRESS AND SUBSEQUENT IN VITRO EMBRYO PRODUCTION IN THE PIG

Fertilization is a crucial step for successful reproduction and can be negatively influenced by stressful situations. It is generally accepted that stress affects reproduction, altering the endocrine profile of the female. An altered hormonal environment where the oocyte is developing could affect critical processes such as fertilization. Using a mixed in vivo–in vitro system, we assessed the ability of the oocyte to undergo fertilization and early development after exposure to blood plasma from sows that had experienced simulated stress through repeated injections of adrenocorticotropic hormone (ACTH) before ovulation (known concentrations of cortisol and reproductive hormones as well as exact ovulation time assessed by ultrasonography). Oocytes (n = 926, 7 replicates) collected from abattoir ovaries were matured in TCM-199 with BSA supplemented with hormones (10 IE mL–1 of pregnant mare serum gonadotropin and 5 IE mL–1 of hCG) and insulin-transferrin-selenium (5 μL mL–1) for 24 h, followed by 22 h without supplements. During IVF, gametes were exposed to 10% of pooled plasma (n = 3 per treatment) collected approximately 1 h before ovulation from ACTH-treated sows (A group), nontreated control sows (C group), or media with BSA (B group) for 24 h. Fresh semen was added at 5 × 105 cells mL–1. Afterward, the remaining cumulus cells and sperm were removed from oocytes by vortexing (1 min), and presumptive zygotes were placed in culture medium (porcine zygote medium). Cleavage rate was assessed at 48 h post-insemination (hpi) and the embryos (n = 433, 7 replicates) were cultured up to Day 7 and stained with Hoechst 33342 (10 μg mL–1) to count the total number of nuclei. In addition, non-cleaved oocytes were stained at 48 hpi with Hoechst to assess sperm-zona binding. Binding to the zona was assessed only in oocytes found to be matured. Statistical analysis was done using Kruskal-Wallis ANOVA and the Mann-Whitney U test. The number of spermatozoa bound to the zona pellucida was higher in the B group, and binding was notably negatively affected in the ACTH group (0.43 ± 0.18, 35.93 ± 2.50, and 3.44 ± 1.04 for the A, B, and C group, respectively; P &lt; 0.001). Cleavage rate (over total number of presumptive zygotes) in the A group (30.71 ± 3.76%) was significantly lower than in the control groups (59.93 ± 4.0 and 52.2 ± 5.31% for the B and C group, respectively; P &lt; 0.01). Blastocyst rate expressed over the total number of embryos was reduced in the A group (9.40 ± 5.20%) compared with the controls (27.10 ± 5.79 and 25.66 ± 5.28% in the B and C group, respectively; P &lt; 0.05). However, no differences were found in the total number of nuclei in the blastocysts. The results suggest that fertilization is a sensitive event that could be negatively influenced by stress, subsequently affecting early embryo development. A reduced number of spermatozoa attached to the zona and a lower number of embryos and lower blastocyst development were observed in the simulated-stress group. Further studies would help to elucidate which (in the oocyte, spermatozoon, or both) mechanisms are being affected by ACTH-simulated stress around fertilization. Data are expressed as mean ± SEM. Funded by Formas.

Follicular Persistence Induced by Adrenocorticotropic Hormone Administration in Goats

The objective of the present study was to examine the effect of adrenocorticotropic hormone (ACTH) administration on the induction of persistent cystic follicle in the goat in order to establish a method to experimentally induce cystic follicle. Four cross-bred goats were intramuscularly administered ACTH at 0.78 and 6.25 µg/10 kg twice a day from Days 15 to 21 (Day 0 was defined as the day of last estrus). Follicular status in the ovary was monitored by ultrasound examination. The plasma concentrations of estradiol, progesterone and cortisol were measured. Treatment with ACTH at the 0.78 and 6.25 µg/10 kg levels caused persistent follicles (> 10 days delay from the expected ovulation date) in 50% of the goats in both treatment groups. In those animals, ovulation occurred 17 and 27 days and 11 and 12 days after the expected days in the 0.78 and 6.25 µg/10 kg groups, respectively. The maximum follicle diameters were 10 and 9 mm in the 0.78 and 6.25 µg/10 kg ACTH groups, respectively. In the control group, the estradiol concentration increased on Day 18 and remained at a high level for a few days. However, such an increase was not seen in both ACTH groups. The estradiol concentration increased gradually from Days 21 to 27 in the 6.25 µg/10 kg ACTH group. These results suggest the possibility that ACTH induces persistent follicles in goats, which may be related to the delay of the onset of estradiol secretion followed by its maintenance at a high concentration.