Intact Monkeys Research Articles

Modification of the number and phenotype of striatal dopaminergic cells by carotid body graft

In non-human primates, striatal tyrosine hydroxylase-immunoreactive (TH-ir) cells are increased in number after dopamine depletion and in response to trophic factor delivery. As carotid body cells contain the dopaminotrophic glial cell line-derived neurotrophic factor (GDNF), we evaluated the number, morphology and neurochemistry of these TH-ir cells, in the anterior and posterior striatum of five monkeys treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) which received a graft of carotid body cell aggregates (CBCA) (n = 3) or sham surgery (n = 2), and six MPTP-monkeys that were sacrificed 6 months and 3 years after the last MPTP dose [MPTP I (n = 3) and MPTP II (n = 3), respectively]. Three intact monkeys served as controls. A disability rating scale was used for the assessment of parkinsonism in all lesioned animals, both before and after surgery. For the neurochemical examination, tissue sections were double-labelled with antibodies to TH, dopamine transporter, dopa decarboxylase-67, vesicular monoamine transporter 2, glutamic acid decarboxylase -67, calbindin, parvalbumin, calretinin, neuronal nitric oxide synthase and GDNF. Only animals receiving CBCA graft showed a moderate but significant recovery of parkinsonism that persisted 12 months after the graft. The grafted striatum contained the greatest TH-ir cell density (120.4 +/- 10.3 cells/100 mm2), while the control striatum displayed the lowest (15.4 +/- 6.8 cells/100 mm2), and MPTP I, MPTP II and sham-operated monkeys showed a similar intermediate value (66.1 +/- 6.2, 58.3 +/- 17.2 and 57.7 +/- 7.0 cells/100 mm2, respectively). In addition, in the post-commissural striatum, only CBCA graft induced a significant increase in the TH-ir cell density compared to control animals (47.9 +/- 15.9 and 7.9 +/- 3.2, respectively). Phenotypically, TH-ir cells were striatal dopaminergic interneurons. However, in the grafted animals, the phenotype was different from that in control, MPTP and sham-operated monkeys, with the appearance of TH/GDNF-ir cells and the emergence of two TH-ir subpopulations of different size as the two main differentiating features. Our data confirm and extend previous studies demonstrating that striatal CBCA grafts produce a long-lasting motor recovery of MPTP-monkeys along with an increase in the number and phenotype changes of the striatal TH-ir interneurons, probably by the action of the trophic factors contained in carotid body cells. The increased number of striatal TH-ir cells observed in the grafted striatum may contribute to the improvement of parkinsonism observed after the graft.

Regulation of circulating leptin and its soluble receptor during pubertal development in the male rhesus monkey (Macaca mulatta)

In humans, circulating leptin levels are low in early childhood and rise until puberty, whereas the reverse occurs for the soluble leptin receptor (sOB-R). In women, leptin remains high and sOB-R remains low, but in men leptin declines after adolescence and sOB-R increases. These observations suggest that leptin may regulate the production of sOB-R, and that the increased testosterone in adolescent boys may be responsible for the gender differences in leptin and sOB-R. To test this hypothesis, leptin was administered continuously to agonadal juvenile male monkeys for 16 days. No change in sOB-R was observed. Intact juvenile male monkeys were given pulsatile doses of gonadotropins for a period of 7 weeks to induce precocious puberty and assess the effect on plasma testosterone, leptin, and sOB-R. By 4 weeks testosterone had reached adult levels. No changes were observed in leptin, but by week 4, sOB-R was higher than pretreatment values and remained higher at week 7. These data suggest that leptin may not play a significant role in regulating the production of sOB-R and that gender differences in sOB-R in humans may be driven by the increased production of testosterone at puberty in males.

Brain-computer interfaces: communication and restoration of movement in paralysis.

The review describes the status of brain-computer or brain-machine interface research. We focus on non-invasive brain-computer interfaces (BCIs) and their clinical utility for direct brain communication in paralysis and motor restoration in stroke. A large gap between the promises of invasive animal and human BCI preparations and the clinical reality characterizes the literature: while intact monkeys learn to execute more or less complex upper limb movements with spike patterns from motor brain regions alone without concomitant peripheral motor activity usually after extensive training, clinical applications in human diseases such as amyotrophic lateral sclerosis and paralysis from stroke or spinal cord lesions show only limited success, with the exception of verbal communication in paralysed and locked-in patients. BCIs based on electroencephalographic potentials or oscillations are ready to undergo large clinical studies and commercial production as an adjunct or a major assisted communication device for paralysed and locked-in patients. However, attempts to train completely locked-in patients with BCI communication after entering the complete locked-in state with no remaining eye movement failed. We propose that a lack of contingencies between goal directed thoughts and intentions may be at the heart of this problem. Experiments with chronically curarized rats support our hypothesis; operant conditioning and voluntary control of autonomic physiological functions turned out to be impossible in this preparation. In addition to assisted communication, BCIs consisting of operant learning of EEG slow cortical potentials and sensorimotor rhythm were demonstrated to be successful in drug resistant focal epilepsy and attention deficit disorder. First studies of non-invasive BCIs using sensorimotor rhythm of the EEG and MEG in restoration of paralysed hand movements in chronic stroke and single cases of high spinal cord lesions show some promise, but need extensive evaluation in well-controlled experiments. Invasive BMIs based on neuronal spike patterns, local field potentials or electrocorticogram may constitute the strategy of choice in severe cases of stroke and spinal cord paralysis. Future directions of BCI research should include the regulation of brain metabolism and blood flow and electrical and magnetic stimulation of the human brain (invasive and non-invasive). A series of studies using BOLD response regulation with functional magnetic resonance imaging (fMRI) and near infrared spectroscopy demonstrated a tight correlation between voluntary changes in brain metabolism and behaviour.

Population pharmacokinetic modeling for enterohepatic recirculation in Rhesus monkey

Enterohepatic recirculation (EHR) occurs via biliary excretion and intestinal reabsorption of a drug. Drug recycling through EHR can lead to a change in pharmacokinetic (PK) properties, such as reduced clearance (CL), extended half-life ( T 1/2) and increased plasma exposure (AUC). As a result, EHR may prolong the pharmacological effect of drugs. In the present study, the compound (Cpd A) was found to exhibit EHR in Rhesus monkeys associated with a reduction in CL (from 3.8 to 0.33 L h −1, IV; from 2.3 to 0.4 L h −1, PO), and an increase in T 1/2 (from 0.9 to 18 h, IV) and in AUC (from 1.5 to 17.4 μg h/mL, IV; from 2.8 to 16.3 μg h/mL, PO), by comparing the PK in the monkeys via the interruption of EHR (bile-duct cannulation) with that in the intact monkeys. A population four-compartment model was constructed based on recirculation loops incorporating all possible inputs (bile secretion, a lag-time model for gall bladder emptying, routes and amounts of a single dose administration) to fully evaluate the EHR of Cpd A. The plasma concentrations versus time profiles predicted from the model had a good fit to the values observed in the subjects and were further simulated with 90% confidence interval to demonstrate its utility. Thus, the model could be applied as a useful tool to evaluate the drugs or compounds that undergo EHR in different species.

Effects of anesthesia upon18F-FDG uptake in rhesus monkey brains

The kinetics of 18F-fluorodeoxyglucose (18F-FDG) in the monkey brain were monitored, and comparisons were made between the conscious state and when under ketamine and pentobarbital anesthesia. Rhesus monkeys were intravenously injected with 18F-FDG and followed by 60 min of PET scanning. In the conscious state, the 18F-FDG concentration reached a plateau 5 min after intravenous injection. Under ketamine anesthesia, the 18F-FDG concentration gradually increased with time in all monitored regions. At 60 min after injection, the concentration in the striatum was about 3.2 times greater than that in the conscious state, and about 4.5 times greater in the cerebral cortex. Under pentobarbital anesthesia, the 18F-FDG concentration in the occipital cortex was slightly lower. These findings demonstrate that 18F-FDG concentration in the monkey brain is significantly affected by anesthesia. The results also imply the existence of a short-term regulation mechanism for hexokinase activity in intact monkey brain.

A Unilateral Section of the Corticospinal Tract at Cervical Level in Primate Does Not Lead to Measurable Cell Loss in Motor Cortex

The effects of a unilateral interruption of the dorsolateral funiculus at cervical level on the survival of neurons in the motor cortex were investigated in macaque monkeys. The lesion was made on the left side at the transition region between the 7(th) and 8(th) cervical segments, above the motoneurons controlling hand muscles. As a result, the homolateral hand became paretic, although an incomplete recovery of manual dexterity took place during 2 months post-lesion. A quantitative anatomical assessment of pyramidal neurons in layer V was performed in the hindlimb area of the primary motor cortex and in the supplementary motor area (SMA proper). The pyramidal neurons were visualized using the marker SMI-32 and thus included the subpopulation of corticospinal neurons. These quantitative data demonstrated that the vast majority of the axotomized corticospinal (CS) neurons did not degenerate. Rather, their somata shrank, compared to the opposite hemisphere or to intact monkeys. This conclusion is in contrast to some previous studies in monkeys that argued for a substantial degeneration of motor cortex neurons as a result of transection of the corticospinal tract; yet in agreement with others that concluded the survival of most CS neurons. The survival of the majority of CS axotomized neurons is also consistent with the observation of numerous CS axons 1 mm above the cervical hemisection.

Norethisterone enanthate has neither direct effect on the testis nor on the epididymis: A study in adult male cynomolgus monkeys (Macaca fascicularis)

Norethisterone enanthate (NETE) is currently evaluated in trials of hormonal male contraception. NETE acts suppressing gonadotropin secretion. In this study we analysed whether NETE has a direct effect on the testis and/or on the epididymis in adult monkeys. NETE was given to cynomolgous monkeys in which endogenous gonadotropin secretion was suppressed by a GnRH antagonist (Cetrorelix), and replaced by injecting recombinant human FSH and human chorionic gonadotropin (hCG). We postulated that, if NETE has a direct effect on spermatogenesis and/or epididymal function, some changes in testicular histology, sperm motility and/or morphology should be found to occur soon after exposure to NETE. Fifteen adult intact male monkeys were allocated to three groups and treated for a 38 day period. Group I received GnRH antagonist, recombinant FSH, hCG and NETE while group II received a regime identical to group I without NETE and group III received only NETE and vehicle. Ejaculates, body weight, testicular volume and hormones were evaluated. Testicular biopsies were taken before and after treatment. There was a similar pattern of serum FSH and testosterone in group I and II throughout the study period, whereas serum FSH and testosterone dropped below the limit of detection in group III. Testicular volume and the proportion of tubuli exhibiting elongated and round spermatids was significantly decreased in group III. No significant differences were found between group I and group II in any parameters measured. The forward progression of sperm was not affected by NETE treatment. The low percentages of grade c sperm indicated no sign of NETE-induced hyperactivation. No changes in the acrosome gros morphology were detected among the different treatment groups. These results suggest that short-term NETE treatment has neither direct effects on the testis nor on the epididymis in this non-human primate model.

The Putative Tumor Suppressor Deleted in Malignant Brain Tumors 1 Is an Estrogen-Regulated Gene in Rodent and Primate Endometrial Epithelium

Deleted in malignant brain tumors 1 (DMBT1) is a candidate suppressor of malignancies of the brain, lung, gut, and breast. We have been studying gene expression in the uterus in the presence of estrogens and their antagonists. Here, we show that DMBT1 RNA levels are robustly increased by estrogen treatment in the uteri of ovariectomized monkeys and rats. In monkeys, the progestin antagonist mifepristone inhibits estrogen-dependent uterine proliferation. As determined by a microarray experiment and quantitative analysis of RNA levels, mifepristone inhibited estrogenic induction of DMBT1. DMBT1 was not expressed in intact monkeys that were treated with a gonadotropin agonist to suppress steroidogenesis. An in vitro transfection study with human DMBT1 promoter constructs showed that an Alu site approximately 3000 nucleotides upstream of the gene mediates estrogenic regulation. Surprisingly, the estrogen antagonists tamoxifen, raloxifene, and ICI 182,780 also induced gene expression via this Alu site. Rodents represent a more convenient model system for studying uterine biology than monkeys. In rats, uterine DMBT1 RNA levels were dramatically up-regulated by estrogen. Consistent with the transfection study, tamoxifen and raloxifene increased DMBT1 RNA levels in vivo, but ICI 182,780 inhibited an estrogen-induced increase. Immunohistochemical studies showed that DMBT1 is specifically induced in glandular and luminal epithelia of the rat endometrium. Our experiments establish that DMBT1 is an estrogen-responsive gene with a possible role in endometrial proliferation or differentiation, and they have implications for the putative tumor suppressive and mucosal protective functions of DMBT1 in the uterus.

Movement of cysteine in intact monkey lenses: the major site of entry is the germinative region

Monkey lenses were incubated with 35S- l-cysteine for various times and the movement of label within the lens followed by autoradiography. Cysteine appeared to enter primarily at the germinative region of the lens. No evidence was found for major transport through either the anterior or posterior faces of the lens. The movement of cysteine within different parts of the lens was followed over time. The data suggest that, for cysteine, the major pathway for transport within the lens involves entry at the germinative region followed by movement along the fibre cells. The data were consistent with orthogonal movement across the fibres in the equatorial plane but little or no movement across the fibres at the anterior pole or posterior faces of the lens. Such a scenario is in accord with the distribution of connexons, indicating that this pattern of entry may also be observed for other small molecules. The finding of high permeability at the lens germinative region is in accord with the anatomy of the eye, since this is the lens surface in contact with the posterior chamber. Thus, cysteine secreted by the ciliary body into the aqueous humor would come into contact initially with the region of the lens best able to absorb this amino acid. Although this aspect was not addressed in the current study, the same phenomenon may also be observed with other lens nutrients.

Impairment of context-adapted movement selection in a primate model of presymptomatic Parkinson's disease.

The MPTP model allows the presymptomatic stage of parkinsonism to be studied in primates and hence specific behavioural manifestations of moderate nigrostriatal denervation to be identified. On the basis of the physiological literature, we hypothesized that depletion of striatal dopamine could impair the selection of context-relevant habits. To examine this hypothesis, we trained three African green monkeys to perform a simple reach-and-grasp task, including three contexts differing only in terms of the presence and position of transparent obstacles. At the end of training, the analysis of reaching trajectories showed that intact monkeys had built a repertoire of movements, from which they could select the appropriate one depending on the context. In the course of MPTP intoxication (0.3-0.4 mg/kg every 4-5 days) and before parkinsonian motor symptoms appeared, the reaction time (RT), movement time (MT) and variability of reaching trajectories increased in all monkeys. Frequently, the initial direction was not adapted to the context, and consequently the movement was either corrected online or restarted under visual assistance. These non-adapted trajectories appeared to be the main reason for the increase in both RT (because of difficulty in selecting) and MT (because of the need to make corrections). These observations indicate that moderate MPTP-induced dopamine depletion results in a deficit in the selection of context-adapted movement, which is compensated by corrections using either proprioceptive or visual feedback. Similar behavioural disorders might therefore occur in the presymptomatic stage of human Parkinson's disease.

Distribution and binding parameters of GABA A receptors in the thalamic nuclei of Macaca mulatta and changes caused by lesioning in the globus pallidus and reticular thalamic nucleus

Ascending output from the basal ganglia to the primate motor thalamus is carried by GABAergic nigro- and pallido-thalamic pathways, which interact with intrinsic thalamic GABAergic systems represented in primates by local circuit neurons and axons of the reticular thalamic nucleus. Disease-triggered pathological processes in the basal ganglia can compromise any of these pathways either directly or indirectly, yet the effects of basal ganglia lesioning on its thalamic afferent-receiving territories has not been studied in primates. Two GABA A receptor ligands, [ 3H]muscimol and [ 3H]flunitrazepam, were used to study the distribution and binding properties of the receptor in intact monkeys, those with kainic acid lesions in the globus pallidus, and those with ibotenic acid lesions in the reticular nucleus using quantitative autoradiographic technique on cryostat sections of fresh frozen brain tissue. In control monkeys the binding affinities for [ 3H]muscimol averaged 50 nM in the thalamic nuclei and 86 nM in the basal ganglia while the binding densities varied (maximum density of binding sites [Bmax] range of 99.4–1000.1 fmol/mg of tissue). Binding affinities and Bmax values for [ 3H]flunitrazepam averaged 2.02 nM and 81–113 fmol/mg of tissue, respectively. Addition of 100-μM GABA increased average affinity to 1.35 nM whereas Bmax values increased anywhere from 1–50% in different nuclei. Zolpidem (100 nM) decreased binding by 68–80%. Bmax values for both ligands were decreased at the two survival times in both medial and lateral globus pallidus implying involvement of both nuclei in the lesion. Statistically significant, 40% decrease ( P=0.055) of Bmax for [ 3H]muscimol was observed in the ventral anterior nucleus pars densicellularis (VAdc, the main pallidal projection territory in the thalamus) 1 week after globus pallidus lesioning and a 36% decrease ( P=0.017) 4 months post-lesioning. In contrast, [ 3H]flunitrazepam Bmax values in the VAdc of the same animals were increased by 23% ( P=0.021) at 1 week and 28% ( P=0.005) 4 months postlesion, respectively. One week after the reticular nucleus lesioning, the binding densities of [ 3H]muscimol and [ 3H]flunitrazepam were decreased in the thalamic nuclei receiving projections from the lesioned reticular nucleus sector by approximately 50% ( P<0.05) and 10–33% ( P<0.05), respectively. The results suggest that different GABA A receptor subtypes are associated with different GABAergic systems in the thalamus which react differently to deafferentation.

Immunohistochemical investigation on the pattern of vimentin expression in regenerated and intact monkey and human periodontal ligament

The expression of vimentin is well documented in the intact animal and human periodontal ligament (PDL), but there is limited information on the pattern of vimentin expression in the regenerated PDL. The aim of the present study was to investigate the pattern of vimentin expression in the regenerated and intact monkey and human PDL. A total of 12 chronic recession-type defects were created in three monkeys ( Macaca fascicularis) and treated either with guided tissue regeneration (GTR), or with an enamel matrix protein derivative (EMD). After 5 months, the animals were sacrificed and specimens containing the defects and surrounding tissues were dissected free, decalcified in EDTA and embedded in paraffin. Sections were labelled immunohistochemically by using monoclonal antibody against vimentin (VIM 3B4). Twelve patients, each of whom displayed one deep intrabony defect scheduled for extraction were treated with GTR, EMD or combination of EMD+natural bone mineral (NBM). Following a healing period of 6 months, the teeth were extracted “en block” and immunohistochemically analysed according to the same protocol as described in monkeys. The results revealed that in both monkeys and humans the newly formed PDL was labelled similarly for vimentin to the intact (non-treated) PDL. In all specimens, the newly formed PDL was in continuation with the intact parts of PDL, thus suggesting that the mesenchymal cells capable of regenerating the attachment apparatus may have their origin in the intact PDL. In conclusion, the present findings indicate that (a) the reformed PDL displayed a similar expression of vimentin to the intact (original) PDL, and (b) the cells capable of regenerating new PDL and new cementum appear to be of mesenchymal origin and their source may be in the intact PDL.

Effect of enucleation on the expression of c-Fos protein in the supraoptic nucleus of the Japanese monkey ( Macaca fuscata)

The aim of this study was to examine the effects of one eye enucleation on the expression of c-Fos protein in the hypothalamus of the Japanese monkey ( Macaca fuscata). Compared with an intact monkey, significantly increased numbers of c-Fos positive neurons were observed in the supraoptic nuclei on both sides at 1 h after eye enucleation. This maximal c-Fos expression then started to decrease at 3 h after eye enucleation. Furthermore, by a dual-labeled immunocytochemical study, the c-Fos immunoreactivity was found mainly in the vasopressinergic but not in the oxytocinergic neurons within the supraoptic nucleus. These results suggest that vasopressinergic but not oxytocinergic neurons within the supraoptic nucleus may have critical roles in the stimulation of this nucleus in response to eye enucleation.

Cerebellar influences on saccade plasticity.

Inaccurate saccades adapt to become more accurate. In this experiment the role of cerebellar output to the oculomotor system in adapting saccade size was investigated. We measured saccade adaptation after temporary inactivation of saccade-related neurons in the caudal part of the fastigial nucleus which projects to the oculomotor brain stem. We located caudal fastigial nucleus neurons with single unit recording and injected 0.1% muscimol among them. Two monkeys received bilateral injections and two monkeys unilateral injections. Unilateral injections made ipsiversive saccades hypermetric (gains >1.5) and contraversive saccades hypometric (gains approximately 0.6). Bilateral injections made both leftward and rightward saccades hypermetric (gains >1.5). During unilateral inactivation neither ipsiversive nor contraversive saccade size adapted after approximately 1,000 saccades. During bilateral inactivation, adaptation was either small or very slow. Most intact monkeys completely adapt after approximately 1,000 saccades to similar dysmetrias produced by intrasaccadic target displacement. After the monkeys receiving bilateral injections made >1,000 saccades in each horizontal direction, we placed them in the dark so that the muscimol dissipated without the monkeys receiving visual feedback about its saccade gain. After the dark period, 20-degree saccades were adapted to be 12% smaller, and 4-degree saccades to be 7% smaller. We expect this difference in adaptation because during caudal fastigial nucleus inactivation, monkeys made many large overshooting saccades and few small overshooting saccades. We conclude from these results that: (1) caudal fastigial nucleus activity is important in adapting dysmetric saccades; and (2) bilateral caudal fastigial nucleus inactivation impairs the relay of adapted signals to the oculomotor system, but it does not stop all adaptation from occurring.

Learning motivational significance of visual cues for reward schedules requires rhinal cortex

The limbic system is necessary to associate stimuli with their motivational and emotional significance. The perirhinal cortex is directly connected to this system, and neurons in this region carry signals related to a monkey's progress through visually cued reward schedules. This task manipulates motivation by displaying different visual cues to indicate the amount of work remaining until reward delivery. We asked whether rhinal (that is, entorhinal and perirhinal) cortex is necessary to associate the visual cues with reward schedules. When faced with new visual cues in reward schedules, intact monkeys adjusted their motivation in the schedules, whereas monkeys with rhinal cortex removals failed to do so. Thus, the rhinal cortex is critical for forming associations between visual stimuli and their motivational significance.

Antiproliferative effects of progesterone antagonists and progesterone receptor modulators on the endometrium

Progesterone antagonists (PAs, antiprogestins) can modulate estrogenic effects in various estrogen-dependent tissues. These modulatory effects are complex and depend on species, tissue, type of compound, dose, and duration of treatment. In non-human primates, PAs, including mifepristone, ZK 137 316 and ZK 230 211, inhibit endometrial proliferation and induce amenorrhea. When administered chronically at relatively low doses, these compounds block the mitotic activity of endometrial epithelium and induce stromal compaction in a dose-dependent manner in both spayed and intact monkeys at high estradiol concentrations. These effects were accompanied by an atrophy of spiral arteries. The antiproliferative effects were endometrium-specific, since the estrogenic effects in the oviduct and vagina were not inhibited by PAs. Similar endometrial antiproliferative effects were also found after treatment with the progesterone receptor modulator (PRM), mesoprogestin J1042. The endometrial antiproliferative effects of PAs, particularly within the endometrial glands, were also observed in spayed rabbits. In spayed rats, however, the PAs did not inhibit, but rather enhanced, various estrogen responses, including endometrial proliferation, pointing to species-specific differences. In conclusion, our studies indicate that both pure PAs and PRMs selectively inhibit estrogen-dependent endometrial proliferation in the primate endometrium without affecting estrogenic response in other estrogen-dependent tissues or inducing unscheduled bleeding. Our studies indicate that the spiral arteries, which are unique to the primate endometrium, are the primary targets that are damaged or inhibited by PAs and PRMs. The damage to these unique vessels may underlay the paradoxical, endometrium-specific, antiproliferative effects of these compounds. Hence, the properties of PAs and PRMs (mesoprogestins) open up new applications in gynecological therapy and hormone replacement therapy.

Testosterone modulates growth hormone secretion at the hypothalamic but not at the hypophyseal level in the adult male rhesus monkey

We investigated a possible modulation of growth hormone (GH) secretion by testosterone by measuring the growth hormone releasing hormone (GHRH)-stimulated and N-methyl-d,l-aspartic acid (NMA)-induced GH secretion in adult rhesus monkeys. Intact, orchidectomized and testosterone-substituted (testosterone enanthate 125 mg/week, i.m. for 5 weeks) orchidectomized monkeys (n=5) were used in the study. GHRH (25 microg/kg body weight) or NMA (15 mg/kg body weight) was infused through a Teflon cannula implanted in the saphenous vein. Sequential blood samples were collected 30-60 min before and 60 min after the injection of the neurohormone or the drug at 10-20-min intervals. All bleedings were carried out under ketamine hydrochloride anaesthesia (initial dose 5 mg/kg body weight i.m., followed by 2.5 mg/kg at 30-min intervals). The plasma concentrations of GH, testosterone and oestradiol (E(2)) were determined by using specific assay systems. Administration of GHRH elicited a significant increase in GH secretion in all three groups of animals. There was no significant difference in the responsiveness of pituitary somatotrophs to exogenous GHRH challenges between intact and orchidectomized monkeys and testosterone replacement in orchidectomized animals did not significantly alter the GHRH-induced GH response. The responsiveness of hypothalamic GHRH neurones apparently did undergo a qualitative change after orchidectomy, as GH response to NMA was less in orchidectomized animals than in intact monkeys. The responsiveness of GHRH neurones to exogenous NMA was restored and even potentiated when orchidectomized monkeys were treated with testosterone. Taken together, these findings suggest that testosterone does not affect the sensitivity of the pituitary somatotrophs to GHRH but stimulates the secretion of GH by modulation of the NMDA drive to GHRH neurones.

Ultrastructure of langur monkey epididymidis prior to and following vasectomy and vasovasostomy.

The ultrastructural changes in langur monkey epididymis prior to and following vasectomy or vasovasostomy were studied. The epididymal epithelium of the intact langur monkey was found to consist mainly of principal cells and basal cells and frequently apical or mitochondria rich cells were found. Besides these cells intraepithelial lymphocytes were also a consistent feature of the epididymal epithelium. Principal cells identified by means of the tuft of the stereocilia on their apical surface, bear well developed Golgi bodies, endoplasmic reticulum, vesicles, vacuoles and multivesicular bodies. This suggests their active involvement in absorption and secretion. Basal cells present at the base of the lamina bear a few cellular organelles and strong interdigitations with the adjacent cells. Apical or mitochondria rich cells were characterized by clusters of mitochondria in the apical region of the cell and few microvilli on their apical surface. Lymphocytes with a large nucleus and a pale rim of cytoplasm were also found at the base of the epithelium. Secretory and absorptive functions of principal cells of the epididymal epithelium were found to be increased after vasectomy, as indicated by bulging of the apical portion of the principal cells and membrane bound structure in the lumen. An extensive increase in the number of lysosomes, vesicles and vacuoles was also observed. An increase in the number of macrophages with spermatozoa remnants in the lumen of epididymis suggests that the principal mechanism for spermatozoa disposal following vasectomy is intraluminal endocytosis by macrophages. Changes following vasectomy persisted in vasovasostomized animals even after 12 months of recanalization, which may contribute to the failure of functional reanastomosis.

Monkey pulpal response and microtensile bond strength beneath a one-application resin bonding system in vivo

Objectives: The aim of this in vivo study was to investigate the biocompatibility and microtensile bond strength of a one-application resin bonding system. Methods: Class V cavities were prepared on the facial surfaces of 36 intact monkey teeth, and the cavities were restored with an experimental one-application resin bonding system (TOF-1; Tokuyama Corp., Tokuyama, Japan) and a hybrid resin composite (PALFIQUE ESTELITE; Tokuyama Corp., Tokuyama, Japan). Histopathological changes of the restored teeth were evaluated at 3, 30 and 90 days after operation ( N=10). Microtensile bond tests were performed at 3 and 90 days after operation ( N=10). Results: Only two of 30 pulps showed a slight inflammatory cell infiltration. There were no statistically significant differences in the incidence of slight inflammatory cell infiltration among time periods. Bacterial penetration along the cavity walls could not be detected in any specimen. The mean microtensile bond strength at 3 days after operation was 20.6 MPa, and that at 90 days was 14.9 MPa. Differences in bond strengths between the 3 day specimen and the 90 day specimen were statistically significant ( p<0.05). Conclusions: The one-application resin bonding system exhibited acceptable biologic compatibility to the monkey pulp. Although there were statistically significant differences in bond strengths between the 3 day specimen and the 90 day specimen, this material provided a hermetic seal, eliminating bacterial microleakage.

Estradiol and High‐Dose Dihydrotestosterone Treatment Causes Changes in Cynomolgus Monkey Prostate Volume and Histology Identical to Those Caused by Testosterone Alone

To evaluate the effects of testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) on the regulation of prostate growth and tissue composition, the following study was conducted in a nonhuman primate model. Fifteen adult, long-term castrated cynomolgus monkeys were randomly assigned to receive implants filled with T (0.19 +/- 0.01 g), DHT alone (0.21 +/- 0.01 g), or (99%) DHT + (1%) E2 (0.21 +/- 0.01 g). Prior to and at 4-week intervals during the treatment phase of 252 days, prostate volumes (PV), body weight, ejaculate weight, hormone levels (of T, DHT, and E2), and red blood cell count were measured. Five adult, intact monkeys served as controls for prostate volume and histology. At the end of the study, histological analysis of an ultrasound-guided prostate biopsy was performed. T levels increased significantly in the T group compared with baseline (P < 0.01) and with the DHT and DHT + E2 groups (P < 0.05). Both groups receiving DHT showed higher DHT levels than did animals in the T group (P < 0.001). E2 levels in all groups increased over time (P < 0.05), although significant differences (P < 0.01) could only be detected between the DHT + E2 and the DHT group. Prostate volume in all groups increased (at baseline: T = 1.03 +/- 0.12 ml, DHT = 1.08 +/- 0.15 ml, DHT + E2 = 1.13 +/- 0.09; at day 252: T = 5.83 +/- 1.00, DHT = 4.72 +/- 0.9, DHT + E2 = 5.05 +/- 0.62) over time (P < 0.001), whereas no differences could be detected between the groups. Prostate biopsy could be performed successfully in 15 out of 20 monkeys. Prostate tissue evaluation between the treatment groups and the evaluated intact monkeys revealed no differences in the status of secretory epithelia, nuclear chromatin, excretory vacuoles, interstitial stroma, smooth muscles, and total functional status, whereas the prostate of a long-term castrated monkey showed severe atrophy. Thus, both androgens fully restored prostate volume and ejaculatory function. Highly supraphysiological DHT serum levels are not associated with abnormal volumetric or histological changes of the prostate. Comparing the DHT group with the DHT + E2 group, an additional stimulatory effect of normal or slightly elevated estrogens on the prostate cannot be found in the presence of highly supraphysiological DHT levels.