Female Cynomolgus Monkeys Research Articles

Abstract 12419: Preclinical Safety and Efficacy of a Novel Thrombolytic Agent Administered by Rapid Bolus Injection: Clot Specific Streptokinase (CSSK/SMRX-11)

Introduction: Clot specific Streptokinase (CSSK), also known as SMRX-11, is a novel recombinant protein, produced and purified from a yeast system (Pichia pastoris). Novel fibrin binding domains attached to each end of streptokinase mask the ability of the streptokinase component of SMRX-11 to interact with blood plasminogen. As a result, SMRX-11 remains inactive in the blood circulation and does not convert blood plasminogen into plasmin until the fibrin binding domains are cleaved by clot-bound plasmin. This is in contrast to native Streptokinase (SK), which indiscriminately converts plasminogen into plasmin. The molecular weight of the glycosylated SMRX-11 protein is approximately 80kD, with a specific activity of 80,000 Units/mg of protein. Results. In vitro experiments comparing SMRX-11 to other fibrinolytics (SK and tPA), demonstrated that SMRX-11 lyses experimentally-preformed human blood clots without reducing residual plasma fibrinogen levels. Initial in vitro studies compared the activity of SMRX-11 to lyse human whole blood clots to native Streptokinase (SK) and tissue Plasminogen Activator (tPA). SMRX-11 had comparable clot lysis activity to SK, with tPA eliciting a slightly greater percent clot lysis. Additional in vitro studies determined the relative fibrinogenolytic effects of SMRX-11 versus tPA. SMRX-11 demonstrated superior fibrinogen sparing compared to tPA with minimal consumption of alpha-2 antiplasmin relative to native SK. In vivo, SMRX-11 (0.7-1.4 mg/kg as an iv bolus), lysed experimentally-induced femoral arterial thrombi in cynomolgus monkeys with relatively minor fluctuation in plasma fibrinogen, forearm bleeding time or hemodynamics. In GLP safety studies, treatment with SMRX-11 at doses of 1, 3 and 10 mg/kg/day for 5 consecutive days was generally well-tolerated in male and female cynomolgus monkeys. Thus, the low dose of 1 mg/kg was considered the No-Observed Adverse Effect Level (NOAEL) for daily repeat-dose administration of SMRX-11 administered via bolus i.v. injection in cynomolgus monkeys. Conclusion. SMRX-11 represents a novel pharmacophore with the appropriate preclinical safety and efficacy profile to warrant clinical investigation for acute treatment of myocardial infarction and stroke.

Influence of reproductive hormones on social rank and vulnerability to cocaine reinforcement in female cynomolgus monkeys

Influence of reproductive hormones on social rank and vulnerability to cocaine reinforcement in female cynomolgus monkeys

Anhedonia, depression, anxiety, and craving for opiates in opiate addicts stabilized on oral naltrexone and long acting naltrexone implant

Aims: The present study utilized female cynomolgus monkeys and examined whether estrogen (E2), progesterone (P4) and total testosterone (T) predicted eventual social rank, changed following hierarchy establishment, or was associated with susceptibility to cocaine self-administration. Methods: Blood serum was collected for three months prior to social housing, then for one month once hierarchies were stable. Following stable hierarchy formation, monkeys were trained to respond for food under a fixed-ratio 30 schedule of reinfocement and then given access to ascending doses of cocaine (0.0003–0.1mg/kg/inj). Results: Prior to social housing, E2 exposure across the menstrual cycle was greater in eventual subordinate compared to eventual dominantmonkeys, while therewere no differences in P4 concentrations. After stable hierarchies formed (4 monkeys/pen), E2 in subordinates decreased and differences between groupswere no longer observed. Furthermore, dominant animals had decreases in P4 after hierarchies stabilized, whereas subordinate animals showed increases in P4 following stable social group formation. T concentrations did not predict or differ between ranks; decreases in T concentrations were observed across all ranks following stable hierarchy formation. Subordinate monkeys acquired cocaine self-administration at lower doses than dominant monkeys. This effect could be driven by higher circulating E2 since there was an inverse relationshipbetweenE2concentrations anddoseof cocaine at which acquisition occurred. Conclusions: These results enhance our understanding of an individual’s response to environmental manipulations, such as attainment and occupation of social ranks and between-rank variation in hormone dysregulation as well as their relationship to cocaine self-administration. Such informationwill aid in the developmentof pharmacological treatment strategies for drugaddiction. Financial support: DA017763, DA036973.

Effects of 18-month treatment with bazedoxifene on enzymatic immature and mature cross-links and non-enzymatic advanced glycation end products, mineralization, and trabecular microarchitecture of vertebra in ovariectomized monkeys

Bazedoxifene (BZA) is used for the treatment of post-menopausal osteoporosis. To elucidate changes in collagen, mineralization, and structural properties and their relationship to bone strength after treatment with BZA in ovariectomized (OVX) monkeys, the levels of collagen and enzymatic immature, mature, and non-enzymatic cross-links were simultaneously examined, as well as trabecular architecture and mineralization of vertebrae. Adult female cynomolgus monkeys were divided into 4 groups (n=18 each) as follows: Sham group, OVX group, and OVX monkeys given either 0.2 or 0.5mg/kg BZA for 18months. Collagen concentration, enzymatic and non-enzymatic pentosidine cross-links, whole fluorescent advanced glycation end products (AGEs), trabecular architecture, mineralization, and cancellous bone strength of vertebrae were analyzed. The levels of enzymatic immature and mature cross-links, bone volume (BV/TV), and trabecular thickness (Tb.Th) in BZA-treated groups were significantly higher than those in the OVX control group. In contrast, the trabecular bone pattern factor (TBPf), the structure model index (SMI), the enzymatic cross-link ratio, and the levels of pentosidine and whole AGEs in BZA-treated groups were significantly lower than those in the OVX control group. Stepwise logistic regression analysis revealed that BV/TV, Tb.Th, TbPf, and pentosidine or whole AGEs independently affected ultimate load (model R2=0.748, p<0.001) and breaking energy (model R2=0.702). Stiffness was affected by Tb.Th, enzymatic immature cross-link levels and their ratio (model R2=0.400). Treatment with BZA prevented OVX-induced deterioration in the total levels of immature enzymatic cross-links and AGEs accumulation and structural properties such as BV/TV, Tb.Th, and TbPf, which contribute significantly to vertebral cancellous bone strength.

Abstract 641: Trastuzumab-dolaflexin, a highly potent Fleximer-based antibody-drug conjugate, demonstrates a favorable therapeutic index in exploratory toxicology studies in multiple species

Abstract Antibody-drug conjugates (ADCs) represent a promising drug class that relies on monoclonal antibody recognition of specific cancer-associated antigens for targeted delivery of chemotherapeutic agents. Fleximer® ADCs utilize a polymer-based conjugation platform to enable high drug-antibody ratios (DAR) and significantly greater anti-tumor potency compared to ADCs with DAR 3-4. We previously showed that trastuzumab-dolaflexin (T-dolaflexin), a HER2-targeted ADC comprised of the antibody trastuzumab conjugated to ∼15 proprietary auristatin molecules via Fleximer, has excellent pharmacokinetics and efficacy in mouse models (AACR 2014 Abstract #2645). T-dolaflexin is efficacious at a single dose of 0.67 mg/kg in mouse xenograft models, and achieves prolonged tumor-free survival after a single 2 mg/kg dose in a low HER2-expressing model that is insensitive to ado-trastuzumab emtansine (T-DM1). We sought to test the tolerability of T-dolaflexin in exploratory mouse, rat and non-human primate toxicology studies. Cohorts of 6 mice were treated with a single dose of vehicle or T-dolaflexin at 20, 30 or 40 mg/kg and followed for 21 days. The 20 and 30 mg/kg doses were well-tolerated based on body weight loss and mortality and achieved a therapeutic index (TI) in mouse &amp;gt;40X. Cohorts of 4 female cynomolgus monkeys were treated with a single dose of vehicle or T-dolaflexin at 0.67, 1.34 or 2.68 mg/kg (payload doses 600, 1200 and 2400 μg/m2, respectively). Day 3 necropsy was performed on 2 animals per cohort, with recovery necropsy on Day 22 in the remaining 2 animals. All animals survived until scheduled necropsy with limited body weight loss. There were no test-article related findings on gross pathology. Most notable clinical pathology findings were transaminase elevations (primarily AST), and decreased platelet counts at Day 8. One high dose animal demonstrated laboratory findings consistent with hepatic injury with full recovery by Day 22 and no findings on gross or microscopic pathology at necropsy. There was no evidence of myelosuppression. Microscopic pathology findings were limited, with no test-article related findings in HER2-expressing organs including heart, lungs and GI tract. All doses were considered well-tolerated. Toxicokinetics demonstrated good stability of drug conjugate in plasma with t1/2 ∼5 days (comparable to antibody t1/2) and minimal exposure to free payload. Plasma exposure at the 2.68 mg/kg dose in cyno was several fold higher than mouse exposure at the minimally efficacious dose of 0.67 mg/kg, and was comparable to the mouse exposure associated with prolonged tumor free survival after a single 2mg/kg dose. Trastuzumab-dolaflexin, a highly potent Fleximer-based ADC, demonstrates a favorable TI in tumor models with low HER2 expression where current HER2-directed therapies are inactive. Citation Format: Natalya Bodyak, Alex Yurkovetskiy, Peter U. Park, Dmitry R. Gumerov, Michael DeVit, Mao Yin, Joshua D. Thomas, LiuLiang Qin, Timothy B. Lowinger, Donald A. Bergstrom. Trastuzumab-dolaflexin, a highly potent Fleximer-based antibody-drug conjugate, demonstrates a favorable therapeutic index in exploratory toxicology studies in multiple species. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 641. doi:10.1158/1538-7445.AM2015-641

Synthesis of ([11C]carbonyl)raclopride and a comparison with ([11C]methyl)raclopride in a monkey PET study

IntroductionThe selective dopamine D2 receptor antagonist raclopride is usually labeled with carbon-11 using [11C]methyl iodide or [11C]methyl triflate for use in the quantification of dopamine D2 receptors in human brain. The aim of this work was to label raclopride at the carbonyl position using [11C]carbon monoxide chemistry and to compare ([11C]carbonyl)raclopride with ([11C]methyl)raclopride in non-human primate (NHP) using PET with regard to quantitative outcome measurement, metabolism of the labeled tracers and protein binding. MethodsPalladium-mediated carbonylation using [11C]carbon monoxide, 4,6-dichloro-2-iodo-3-methoxyphenol and (S)-(-)-2-aminomethyl-1-ethylpyrrolidine was applied in the synthesis of ([11C]carbonyl)raclopride. The reaction was performed at atmospheric pressure using xantphos as supporting phosphine ligand and palladium (π-cinnamyl) chloride dimer as the palladium source. ([11C]Methyl)raclopride was prepared by a previously published method. In the PET study, two female cynomolgus monkeys were used under gas anesthesia of sevoflurane. A dynamic PET measurement was performed for 63min with an HRRT PET camera after intravenous injection of ([11C]carbonyl)raclopride and ([11C]methyl)raclopride, respectively, during the same day. The order of injection of the two PET radioligands was changed between the two monkeys. The venous blood sample for measurement of protein binding was taken 3min prior to the PET scan. Binding potential (BPND) of the putamen and caudate was calculated with SRTM using the cerebellum as a reference region. ResultsThe target compound ([11C]carbonyl)raclopride was obtained with 50±5% decay corrected radiochemical yield and 95% radiochemical purity. The trapping efficiency (TE) of [11C]carbon monoxide was 65±5% and the specific radioactivity of the final product was 34±1GBq/μmol after a 50min of synthesis time. The radiochemical yield of ([11C]methyl)raclopride was in the same range as published previously i. e. 50–60% and specific radioactivity of those two batches which were used in the present PET study were 192GBq/μmol and 638GBq/μmol respectively after a synthesis time of 32min. In monkey PET studies, the percentage difference of BPND in putamen was <3% and that in caudate was <9% for the two radioligands. The plasma protein binding was 86.2±0.3% and 85.7±0.6% for ([11C]carbonyl)raclopride and ([11C]methyl)raclopride, respectively. The radiometabolite pattern was similar for both radioligands. ConclusionRaclopride was 11C-labeled at the carbonyl position using a palladium-mediated [11C]carbonylation reaction. A comparison between ([11C]carbonyl)raclopride and ([11C]methyl)raclopride with regard to quantitative PET outcome measurements, metabolism of radioligands and protein binding in monkey was performed. The monkey PET study with ([11C]carbonyl)raclopride showed similar results as for ([11C]methyl)raclopride. The PET studies were performed on 2 subjects.

A surgical technique using the ovarian vein in non-human primate models of potential living-donor surgery of uterus transplantation.

Living donor surgery in organ transplantation should be performed in a minimally invasive manner under conditions that are as safe as possible. The objective of this study is to examine whether the procedure for using the ovarian vein makes donor surgery less invasive in a cynomolgus monkey model of potential living-donor surgery of uterus transplantation. Twenty-two female cynomolgus monkeys aged 6-9 years and with body weights of 3.55 ± 1.28 kg were used in the study. Vessels and tissues surrounding the uterus were dissected while preserving the uterine artery/vein. The deep uterine vein was used as a venous pedicle in four monkeys (Group 1), and the ovarian vein was used instead of the deep uterine vein in 18 monkeys (Group 2). With the uterine artery/vein and deep uterine vein (Group 1) or ovarian vein (Group 2) connected to the uterus, the vaginal canal was cut. The vessels were then clamped to produce a donor surgery model. Surgical time, intraoperative organ and vascular injury were examined in each animal. The average surgical time from laparotomy to clamping of vessels was 230 ± 112 min in all 22 cynomolgus monkeys, and significantly longer in Group 1 (n = 4) than in Group 2 (n = 18) (393 ± 71 vs. 194 ± 84 min, p < 0.05). Surgical time in Group 2 showed a tendency to decrease in animals treated later in the study, with a significantly longer time in the first 10 monkeys compared with the last 8 (253 ± 65 vs. 120 ± 26 min, p < 0.05). All monkeys had no complications, including no injuries to other organs and no unanticipated vascular injury. The procedure using the ovarian vein was less invasive than that using the deep uterine vein in mimicking living-donor surgery in a cynomolgus monkey model of uterus transplantation.

Relationship between estradiol and progesterone concentrations and cognitive performance in normally cycling female cynomolgus monkeys

Preclinical research has demonstrated that cognitive function may be influenced by estradiol (E2) and progesterone (P4) concentrations, although few cognition studies involve normally cycling females. The present study examined cognitive performance in normally cycling female cynomolgus macaques (n=14), a species with similarities to humans in brain organization and a nearly identical menstrual cycle to women. Initial assessments compared cognitive measures to circulating concentrations of E2 and P4 (n=12). Once a relationship was characterized between hormones and cognitive performance, the menstrual cycle was divided into four distinct phases: early follicular (EF), late follicular (LF), early luteal (EL) and late luteal (LL), verified by the onset of menses and serum concentrations of E2 and P4. Concentrations of E2 were highest during the LF phase and P4 concentrations peaked during the EL phase. All monkeys were trained on two cognitive tasks: reversal learning, involving simple discrimination (SD) and reversal (SDR), which measured associative learning and behavioral flexibility, respectively (n=3–4 per phase) and a delayed match-to-sample (DMS) task which assessed working memory (n=11). P4 concentrations were positively correlated with number of trials and errors during acquisition of SD performance, but not during acquisition of the SDR task or maintenance of the reversal-learning task. Across the menstrual cycle, significantly fewer errors were made in the SDR task during the LF phase, when E2 concentrations were high and P4 concentrations low. Working memory, assessed with the DMS task, was not consistently altered based on previously characterized menstrual cycle phases. These findings demonstrate a relationship between P4, E2 and cognitive performance in normally cycling cynomolgus monkeys that is task dependent. Knowledge of these interactions may lead to a better understanding of sex-specific cognitive performance.

Influence of Hormones on Social Rank and Vulnerability to Cocaine Reinforcement in Female Cynomolgus Monkeys

Nonhuman primate (NHP) social groups, which represent a continuum from environmental enrichment in dominant (DOM) monkeys to chronic social stress in subordinates (SUB), have been used to study susceptibility and resilience to human diseases, including drug abuse. We have shown that social rank influences sensitivity to cocaine (COC) in NHPs, although it is not clear how hormone fluctuations may influence rank or how hormones are modified after rank is established. This study used female NHPs and examined whether estrogen (E2), progesterone (P4) or testosterone (T) predicted eventual rank, changed following hierarchy establishment, or was associated with susceptibility to COC self‐administration (SA). Prior to social housing, E2 exposure across the menstrual cycle was greater in eventual SUB compared to eventual DOM; there were no differences in P4. After hierarchies were formed (4 monkeys/pen), E2 exposure across the cycle decreased in SUB and differences between ranks were no longer observed. DOM had decreases in P4 after hierarchies stabilized, whereas SUB showed increases in P4 following rank formation. T concentrations did not predict or differ between ranks; decreases in T concentrations were observed across all ranks following hierarchy formation. When given access to COC (0.0003‐0.1 mg/kg/inj), SUB acquired COC SA at lower doses than DOM. This effect could be driven by higher circulating E2 since an inverse relation between E2 and dose of COC at which SA occurred was observed. These results enhance our understanding of an individual's response to factors related to social environment and their relationship to COC SA. Such information will aid in the development of pharmacological treatment strategies for drug addiction. DA017763, DA036973

MP8-01 THE ESTABLISHMENT OF AN UNDERACTIVE BLADDER MODEL FOLLOWING A HYSTERECTOMY IN MONKEY

INTRODUCTION AND OBJECTIVES: Underactive bladder (UAB) syndrome describes a dysfunctional condition of the bladder where patients are unable to produce an effective voiding contraction. To date a pathophysiological mechanisms are not fully understood. To address this issue, we established on underactive bladder model in monkeys after having a hysterectomy. METHODS: Female cynomolgus monkeys were anesthetized with ketamine chloride and pentobarbital sodium. The uterine corpus was then surgically removed while preserving the uterine cervix. Days after surgery, conscious animals were placed on chairs to monitor natural urination after oral administration of 150 mL water (uroflowmetry, UFM). In addition to this, in order to confirm the detrusor underactivity, pressure flow study (PFS) was conducted in the conscious animals. An in vitro muscle strip study was also performed. RESULTS: Monkeys showed typical detrusor underactivity in PFS. Maximum flow rate (Qmax), average flow rate (Qave) and voided volume per micturition were significantly decreased at 2, 4, 6, 8, 21, 60 days after surgery compared with before surgery. On the contrary, voiding time per micturition was significantly increased. Sham-operated group did not show any changes. In muscle strip studies, there was significant decrease in maximum response to potassium chloride (KCl) in the 2 months group but not in the 2 weeks group compared with the sham group. There was a significant decrease in the response to carbachol (CCh) and electric field stimulation (EFS) in both the 2 weeks and 2 months group. CONCLUSIONS: This is the first report on UAB model following a hysterectomy in monkeys. This model had an acutely neurogenic but chronically neurogenic and myogenic characteristics. It seems to be useful in the pathophysiological elucidation of UAB and have potential for assessment of pharmacotherapy of UAB. Source of Funding: none

Cardiac safety profile of etamicastat, a novel peripheral selective dopamine-β-hydroxylase inhibitor in non-human primates, human young and elderly healthy volunteers and hypertensive patients

The aim of this work was to evaluate the cardiac risk for etamicastat, a peripheral reversible dopamine-β-hydroxylase inhibitor. Etamicastat blocked the hERG current amplitude with an IC50 value of 44μg/ml. Etamicastat had no substantial effects on arterial blood pressure, heart rate and the PR interval in male Cynomolgus monkeys when administered orally up to 90mg/kg. Administered orally at 15 and 45mg/kg/day in female and male Cynomolgus monkey for 91days, etamicastat had no effect on heart rate and the waveform or intervals of the electrocardiogram. At the highest dose level of 45mg/kg, mean plasma concentrations of etamicastat ranged from 1875 to 3145ng/ml on Day 1 and Day 91 of treatment, respectively. The effect of age on the tolerability and pharmacokinetics of etamicastat in elderly (≥65years) and young adult (18–45 years) subjects showed that supine systolic (SBP) and diastolic (DBP) blood pressure, ECG heart rate, PR interval, QRS duration and QTcF interval were not affected following once-daily administration of 100mg/day etamicastat for 7days. In hypertensive patients the decrease of blood pressure tended to be more important in subjects who had received etamicastat (50, 100 and 200mg) than in subjects who had received placebo. No clinically significant out-of-range values in vital signs or ECG parameters, ECG heart rate, PR interval, QRS duration and QTcF interval were observed in hypertensive subjects following once-daily administration of etamicastat for 10days. In conclusion, etamicastat is not likely to prolong the QT interval at therapeutic doses.

Effects of prior cocaine self-administration on cognitive performance in female cynomolgus monkeys.

Cocaine use has been associated with cognitive impairments that may contribute to poor treatment outcomes. However, the degree to which these deficits extend into periods of abstinence has not been completely elucidated. This study tested whether prior experience self-administering cocaine affected acquisition of two cognitive tasks in 16 adult female cynomolgus monkeys. Seven monkeys had previously self-administered cocaine but had not had access to cocaine for 2months at the start of this study. After monkeys were trained to respond on a touchscreen, associative learning and behavioral flexibility were assessed using a stimulus discrimination (SD) and reversal (SDR) task from the CANTAB battery. Performance on this task was monitored over the subsequent 3months. Additionally, working memory was assessed with a delayed match-to-sample (DMS) task. Cocaine-naïve monkeys required fewer total trials and made fewer errors and omissions before acquiring the SD and SDR tasks compared with monkeys who had previously self-administered cocaine; two monkeys in the latter group did not acquire the task. However, this cognitive impairment dissipated over several months of exposure to the task. The number of sessions for touch training and delays required to establish a performance-based curve on the DMS task did not differ between groups. Results suggest that cocaine exposure can impair the ability to learn a novel task requiring behavioral inhibition and flexibility, even after an extended period of abstinence. However, this deficit did not extend to maintenance of the task or to acquisition of a working memory task.

Daily application of alprostadil topical cream (Vitaros) does not impact vaginal pH, flora, or histology in female cynomolgus monkeys.

Topical alprostadil cream (Vitaros) is approved in Canada and Europe for the treatment of erectile dysfunction. To determine the effects on the female urogenital tract with repeated administration of the entire dose (300 μg alprostadil containing 2.5% dodecyl-2-n,n-dimethylaminopropionate hydrochloride), the vaginal pH, flora, and histology were assessed as a model for 100% transference from male to the female during unprotected sexual intercourse. Female cynomolgus monkeys were administered the entire dose of Vitaros for 14 days with a 7-day recovery. Relative to vehicle and placebo cream, the vaginal pH and microflora were determined at baseline and weekly, thereafter. Vaginal biopsies were evaluated at the end of dosing and recovery. All animals were clinically normal for the study duration, and the vaginal pH was consistent between dose groups and the dosing period. Vaginal microflora and histopathology findings of mild inflammation were generally similar across treatment groups. In conclusion, repeated vaginal exposure to Vitaros did not alter the pH, microflora, or histology after 14 daily doses, supporting the safety of Vitaros transference to the female partner.

Effects of Catfish Oil Intervention on Lipid Profile in Female Aged Cynomolgous Monkey (Macaca fascicularis)

Degenerative process is an accumulation of free radicals that can lead to a variety of changes in the cell. This aim of the study to see the effects of different dietary lipid intervention on lipid profile and lipid peroxidation in female aged cynomolgus monkey. Twelve female Cynomolgus monkey (Macaca fascicularis) were randomly divided into 4 dietary groups of three animals. Animals were held in individual cages and placed in the position where they can interact individually. They were given a normal fat diet with 2% cholesterol and 3% of fat from soybean oil and 9% of lipid as beef tallow (BFT), catfish oil (CFO), fermented catfish oil (FCFO) and soybean oil (SBO). Evaluation of their body weights, serum lipid profile and cholesterol content consumption were done monthly except an index of lipid peroxidation were performed before and after 12 weeks intervention. Animal group that consumed the feed more than 82% has body weight gain, such as CFO and BFT. On the other side, animal group that consumed less than 70% has body weight loss, such as FCFO. There was no effect (p > 0.05) of the experimental diets on decreasing triglyceride levels and increasing HDL cholesterol level. Cynomolgous given BFT, CFO, and FCFO diet for 3 months showed signifi cantly increased (p < 0.05) in total cholesterol and LDL cholesterol level, but the monkeys fed with SBO showed lower levels. Statistically, All of diet interventions do not significantly affect on lipid peroxidation in LDL (p < 0.05). In general it can be concluded that catfish oil significantly cause elevated levels of total cholesterol and LDL cholesterol in blood serum, but not in the HDL plasma. DOI : http://dx.doi.org/10.17576/JSKM-2014-1202-03

Effects of Catfish Oil Intervention on Lipid Profile in Female Aged Cynomolgous Monkey (Macaca fascicularis)

Degenerative process is an accumulation of free radicals that can lead to a variety of changes in the cell. This aim of the study to see the effects of different dietary lipid intervention on lipid profi le and lipid peroxidation in female aged cynomolgus monkey. Twelve female Cynomolgus monkey (Macaca fascicularis) were randomly divided into 4 dietary groups of three animals. Animals were held in individual cages and placed in the position where they can interact individually. They were given a normal fat diet with 2% cholesterol and 3% of fat from soybean oil and 9% of lipid as beef tallow (BFT), catfi sh oil (CFO), fermented catfi sh oil (FCFO) and soybean oil (SBO). Evaluation of their body weights, serum lipid profi le and cholesterol content consumption were done monthly except an index of lipid peroxidation were performed before and after 12 weeks intervention. Animal group that consumed the feed more than 82% has body weight gain, such as CFO and BFT. On the other side, animal group that consumed less than 70% has body weight loss, such as FCFO. There was no effect (p > 0.05) of the experimental diets on decreasing triglyceride levels and increasing HDL cholesterol level. Cynomolgous given BFT, CFO, and FCFO diet for 3 months showed signifi cantly increased (p < 0.05) in total cholesterol and LDL cholesterol level, but the monkeys fed with SBO showed lower levels. Statistically, All of diet interventions do not signifi cantly affect on lipid peroxidation in LDL (p < 0.05). In general it can be concluded that catfi sh oil signifi cantly cause elevated levels of total cholesterol and LDL cholesterol in blood serum, but not in the HDL plasma.

Uterine autologous transplantation in cynomolgus monkeys: a preliminary report of 6 case

To evaluate the surgical feasibility of uterine autologous transplantation in female cynomolgus monkeys and explore the effect of microsurgical technique. From May 2011 to March 2014, 6 female cynomolgus monkeys, aged 7 to 12 years, were randomly divided into 2 surgical groups. In group A, gross surgeries were performed with naked eyes. In group B, uterine re-transplantation was performed under 10-time-magnifying microscopy. Anatomical data and operative durations were recorded and analyzed. Viable uterine tissue and vascular patency were observed on trans-abdominal ultrasonography and second-look laparotomy after 3 months. The average uterus retrieval time, average vascular anastomotic time, average perfusion time and average total operative time of group B were 88.7, 147.3 and 320.0 min versus 123.7, 180.7 and 393.7 min in group A. The average perfusion durations were 35.0 and 27.3 min. And there was no inter-group difference. A total of 12 successful vascular procedures (including 6 internal iliac arteries and 6 uterine-ovary veins) of vascular anastomosis were recorded in group B versus 4 cases (including 1 artery and 3 veins) in group A (vein, P > 0.05; artery, P < 0.05). In group A, there was only 1 successful operation but uterus turned necrotic after 4 weeks. In group B, the surgical success rate was 100%. However one animal died due to intestinal obstruction. And in another 2 animals, viable uterine tissue and vascular patency were observed on trans-abdominal ultrasonography and second-look laparotomy. Two survivors resumed cyclicity at days 17 and 50 respectively as a sign of re-established uterine function. This study has demonstrated the feasibility of uterus transplantation by vascular anastomosis in cynomolgus monkeys. And assistance of microsurgical technique can significantly improve the success rate of arterial anastomosis during uterus transplantation.

Long-term cocaine self-administration in female cynomolgus monkeys: Changes in behavior and in dopamine transporters and D2-like receptor availability

Long-term cocaine self-administration in female cynomolgus monkeys: Changes in behavior and in dopamine transporters and D2-like receptor availability

Effect of soy isoflavones on thyroid hormones in intact and ovariectomized cynomolgus monkeys (Macaca fascicularis).

Soy isoflavones are commonly used to alleviate menopause-related symptoms. Postmenopausal women are at increased risk for hypothyroidism, and there are concerns that isoflavones may be detrimental to thyroid health. The aim of this study was to examine the effects of soy protein and isoflavones on thyroid function and the relationship between thyroid function and ovarian function. Adult female cynomolgus monkeys (Macaca fascicularis) were randomized to consume two diets differing only in protein source: casein-lactalbumin (n = 44) or soy protein with isoflavones (n = 41). After 34 months, all animals were ovariectomized via laparotomy. Half of the monkeys from each diet treatment group continued to consume their preovariectomy treatment phase diet (either isolated soy protein [n = 19] or casein-lactalbumin [n = 21]) for an additional 34 months. The remaining animals did not continue their diets and thus were not considered further. Circulating progesterone, triiodothyronine, thyroxine, and thyroid-stimulating hormone were measured at baseline. Thyroid hormones were remeasured during each treatment phase. Dietary soy increased triiodothyronine in preovariectomized monkeys and prevented a decline in thyroxine after surgical menopause (both P's < 0.05). Mean progesterone concentrations were positively correlated with triiodothyronine at baseline in preovariectomized monkeys (P < 0.01). Progesterone levels and triiodothyronine are positively correlated in macaques. Dietary soy increases triiodothyronine in preovariectomized monkeys and prevents a decline in thyroxine after surgical menopause. The outcomes observed in this study suggest that soy protein and isoflavone consumption does not adversely affect-and may even preserve-thyroid function in postmenopausal women.

Serum β-nerve growth factor concentrations in pregnant female, nonpregnant female, and male cynomolgus monkeys.

Serum β-nerve growth factor (NGF) concentrations were determined in pregnant female, nonpregnant female, and male cynomolgus monkeys using a highly selective and sensitive immunoaffinity liquid chromatography-tandem mass spectrometry assay. NGF was significantly higher in pregnant monkeys than in nonpregnant female and male monkeys. NGF increased over pregnancy (mean NGF±SD: 541±448, 1590±520, and 3560±1430 pg/ml during the first, second, and third trimesters, respectively). These data will aid in further understanding the role of NGF during pregnancy.

Foodborne transmission of bovine spongiform encephalopathy to non-human primates results in preclinical rapid-onset obesity.

Obesity has become one of the largest public health challenges worldwide. Recently, certain bacterial and viral pathogens have been implicated in the pathogenesis of obesity. In the present study, we retrospectively analyzed clinical data, plasma samples and post-mortem tissue specimens derived from a risk assessment study in bovine spongiform encephalopathy (BSE)-infected female cynomolgus monkeys (Macaca fascicularis). The original study design aimed to determine minimal infectious doses after oral or intracerebral (i.c.) infection of macaques to assess the risk for humans. High-dose exposures resulted in 100% attack rates and a median incubation time of 4.7 years as described previously. Retrospective analyses of clinical data from high-dosed macaques revealed that foodborne BSE transmission caused rapid weight gain within 1.5 years post infection (β = 0.915; P<0.0001) which was not seen in age- and sex-matched control animals or i.c. infected animals. The rapid-onset obesity was not associated with impaired pancreatic islet function or glucose metabolism. In the early preclinical phase of oral transmission associated with body weight gain, prion accumulation was confined to the gastrointestinal tract. Intriguingly, immunohistochemical findings suggest that foodborne BSE transmission has a pathophysiological impact on gut endocrine cells which may explain rapid weight gain. To our knowledge, this is the first experimental model which clearly demonstrates that foodborne pathogens can induce obesity.