Chronic treatment of cycling rhesus monkeys with low doses of the antiprogestin ZK 137 316: morphometric assessment of the uterus and oviduct

The long-term effects of the antiprogestin ZK 137 316 on reproductive tract morphology in rhesus macaques were investigated. The monkeys were injected daily (i.m.) for five menstrual cycles with vehicle or 0.01, 0.03 or 0.1 mg ZK 137 316/kg body weight. Reproductive tracts (n = 3/ group) were collected during the mid-luteal phase (day 8) of the fifth cycle in the control, 0.01 and 0.03 mg/kg groups, or 6-7 days after the oestradiol peak in the 0.1 mg/kg group. ZK 137 316 treatment resulted in a dose-dependent atrophy of the endometrium, marked by reduced mitotic activity in the glands, compaction of the stroma, degradation of spiral arteries and dilation of veins. There was no effect of ZK 137 316 on myometrial or oviductal weight. Treatment with 0.1 and 0.03 mg/kg, but not 0.01 mg/kg resulted in fully ciliated and secretory oviducts, indicating a dose-dependent blockade of progesterone antagonism of oestrogen-dependent oviductal differentiation. In the endometrium, the suppressive action of progesterone on oestrogen and progestin receptors was also blocked by ZK 137 316 in a dose-dependent manner. However, endometrial atrophy appeared due to inhibition of progesterone action together with a blockade of oestrogen-dependent proliferation. The profoundly suppressed endometrium produced by chronic low-dose ZK 137 316 treatment is unlikely to support implantation. Such treatment may therefore provide a novel contraceptive modality.      

Chronic subcutaneous leptin infusion diminishes the responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis in female rhesus monkeys

The fat derived protein leptin has its anorexic action through a number of neuropeptides including an upregulation of corticotropin releasing hormone (CRH) expression in the hypothalamus. However, the influence of leptin on these neuropeptides may be different during stress. The present study used ovariectomized female rhesus monkeys (n=8) to further define the effect of leptin on HPA responsivity. To accomplish this, we assessed the effects of constant leptin infusion on cortisol and ACTH secretion in both a predictable and unpredictable situation as well as in response to dexamethasone suppression-CRH stimulation test. We hypothesized that leptin would attenuate the increase in cortisol and ACTH to a novel, unpredictable situation and would enhance glucocorticoid negative feedback and diminish the response to CRH. Animals were assessed under control placebo conditions and during a 28 day infusion with recombinant human leptin (6 microg/kg/day, SC). Within each treatment condition, HPA responsivity was assessed during no estradiol replacement and acute estradiol replacement that produced serum concentrations of approximately 40 pg/ml. However, the results indicated that neither estradiol alone or in combination with leptin had any consistent effect on the outcome measures. Compared to the control condition, leptin had no effect on the cortisol diurnal rhythm; however, evening but not morning plasma ACTH concentrations were significantly lower during leptin infusion. In contrast, the response in plasma cortisol and ACTH to an unpredictable situation was significantly attenuated by chronic leptin infusion. Furthermore, leptin enhanced glucocorticoid negative feedback and blunted CRH-induced increase in both cortisol and ACTH. Taken together, these data suggest that in the female monkey, leptin has little effect on basal cortisol. However, when the HPA axis is activated, leptin attenuates the neuroendocrine response by enhancing glucocorticoid negative feedback. These data underscore the potential importance of leptin in maintaining homeostasis through its diverse interaction with the HPA axis.     

Interest in infants by female rhesus monkeys with neonatal lesions of the amygdala or hippocampus

Previous research in our laboratory has shown that damage to the amygdala in neonatal rhesus monkeys profoundly alters behaviors associated with fear processing, while leaving many aspects of social development intact. Little is known, however, about the impact of neonatal lesions of the amygdala on later developing aspects of social behavior. A well-defined phenomenon in the development of young female rhesus monkeys is an intense interest in infants that is typically characterized by initiating proximity or attempting to hold them. The extent to which young females are interested in infants may have important consequences for the development of species-typical maternal behavior. Here we report the results of a study that was designed to assess interest in infants by female rhesus monkeys that received neonatal lesions to the amygdala, hippocampus or a sham surgical procedure. Subjects were first paired with pregnant “stimulus” females to assess social interactions with them prior to the birth of the infants. There were few behavioral differences between lesion groups when interacting with the pregnant females. However, following the birth of the infants, the amygdala-lesioned females showed significantly less interest in the infants than did control or hippocampus-lesioned females. They directed fewer affiliative vocalizations and facial expressions to the mother–infant pair compared to the hippocampus-lesioned and control females. These findings suggest that neonatal damage to the amygdala, but not the hippocampus, impairs important precursors of non-human primate maternal behavior.     

Individuated finger movements of rhesus monkeys: a means of quantifying the independence of the digits

1. Two rhesus monkeys were trained to perform flexion and extension movements of each digit of the right hand and of the wrist. Movements of all five digits and the wrist were monitored simultaneously. During each instructed movement, the instructed digit (or wrist) had the greatest excursion; other, noninstructed digits moved to varying degrees. 2. To assess the degree of independence of the different digits during these movements, I plotted, as a function of the instructed digit's position, the position of each noninstructed digit. The resulting trajectories typically were linear, with consistent slopes from trial to trial. 3. The slopes of these noninstructed digit versus instructed digit trajectories were used to calculate an individuation index for each instructed movement and a stationarity index for each digit. These indexes quantified two different aspects of independence. The individuation index reflects the degree to which other digits remained still during instructed movement of a given digit. The stationarity index reflects the degree to which a given digit remained still whenever it was a noninstructed digit. 4. In accordance with casual observation, thumb flexion and wrist flexion and extension consistently had both high individuation and stationarity and therefore can be said to be independent of the fingers. Although the same cannot be said of the other fingers, the present analysis provides a means of quantifying the degree of independence of these digits as well. 5. Factors are discussed that might contribute to the motion of noninstructed digits and to the trajectory linearity.     

Differences among ovariectomized female rhesus macaques in the display of sexual behavior without and with estradiol treatment

Ovariectomized, estrogen-primed, adult female rhesus macaques were preselected for the display of high (HLP) and low (LLP) levels of sexual performance. Eight sexually vigorous adult males were selected as partners. For 13 consecutive days each month, females received 0, 2.5, 5.0, and 10.0 micrograms of estradiol benzoate (EB) and were tested on Days 12 and 13 of injection. Blood samples were taken before and on the last day of each of the four injection series, and serum levels of estradiol, progesterone, and testosterone were determined with a radioimmunoassay. The HLP females (n = 5) displayed higher levels of receptivity and proceptivity and received more mounts, intromissions, and ejaculations when tested without EB treatment than LLP females (n = 5) did when they were tested with 10.0 micrograms of EB. Female receptivity, proceptivity, and attractiveness cannot be explained by estrogen action alone. The effectiveness of estrogen is limited by the somatic characteristics of the individual on which it acts.     

Some factors influencing the effects of temporary mother-infant separation: some experiments with rhesus monkeys.

Some experiments, reported in detail elsewhere, on the effects of mother-infant separation in rhesus monkeys are here reviewed and compared. They involved 4 groups--one in which mothers were removed for 13 days leaving the infant in the social group; one in which infants were removed; one in which mothers and infants were removed and separated; and one in which mothers and infants were removed but not separated. The nature of separation experience had a profound effect on the infant's response: infants left in a familiar environment while their mothers were removed showed marked but brief 'protest' and then profound 'despair', whilst infants removed to a strange cage showed more prolonged 'protest'. A major factor determining the effects of the separation experience in the weeks following reunion is the degree to which the mother-infant relationship has been disturbed by it. The multiplicity of factors affecting the outcome of a separation experience are discussed.     

Intrauterine release of oestriol in castrated rhesus monkeys induces local but not peripheral oestrogenic effects: a possible approach for the treatment and prevention of Asherman's syndrome.

Two types of oestrogen-medicated intrauterine devices (IUD) were studied in ovariectomized rhesus monkeys. An oestradiol (E2) fibre-wrapped IUD that released E2 at a rate of 3.57 micrograms/cm/day, or an oestriol (E3) fibre-wrapped IUD that releases E3 at a rate of 6.4 micrograms/cm/day, was inserted in eight animals and left in place for 4 weeks. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), E2 and E3 were determined by radioimmunoassay for 1 week before the IUD insertion, during the time the IUD was in place, and for 3 weeks after its removal. Uterine histology was performed at the time of IUD insertion and removal by light and transmission electron microscopy. Both E2 and E3 IUDs induced similar histological changes in the uterus, i.e. four- to five-fold increase in endometrial thickness, a shift of the gland/stroma ratio from 1:4 to 1:1, transformation to a marked pseudostratified epithelium with pronounced coiling of the glands, appearance of subnuclear and luminal secretion and, finally, change from spindle-dense stromal cells to plump eosinophilic cells. Oestradiol fibre-wrapped IUDs produced circulating E2 levels of 150-200 pg/ml during the entire 4 weeks. FSH and LH levels were decreased to an average of 55% and 65% from a castration baseline (P less than 0.001 and P less than 0.05, respectively). Oestriol fibre-wrapped IUDs produced circulating E3 levels of 100-250 pg/ml. However, FSH and LH levels were not altered in this group. The specific local oestrogenic effect of E3-IUDs without affecting the pituitary secretion of gonadotrophins, suggests their possible application in cases in which an exclusively oestrogenic effect at the uterine level, such as in Asherman's syndrome, is desired.     

Effects of maternal mobility on the development of rocking and other behaviors in rhesus monkeys: A study with artificial mothers

Mechanically driven mobile artificial mothers effectively prevented the development of stereotyped body-rocking in rhesus monkeys. Monkeys were maternally separated at birth and assigned to 2 groups. Both groups were placed with surrogates, identical in construction except that for 1 group the surrogate was in motion 50% of the time from 0500 hours to 2400 hours each day, and for the other group the surrogate was stationary. All but 1 of the 10 monkeys raised with stationary artificial mothers developed rocking as an habitual pattern whereas none of the 9 monkeys raised with mobile mothers did so. The data also suggest that emotional responsiveness was reduced in monkeys raised with mobile mothers, compared to monkeys raised with stationary devices.     

Age-related changes of the ciliary muscle in comparison with changes induced by treatment with prostaglandin F2 alpha. An ultrastructural study in rhesus and cynomolgus monkeys

The relationship between individual ciliary muscle cells and the surrounding connective tissue was studied in the eyes of three normal, young (3-4 years) cynomolgus monkeys (Macaca fascicularis), three aged (34-36 years) rhesus monkeys (Macaca mulatta) and seven young (3-7 years) cynomolgus monkeys topically treated with prostaglandin F2 alpha (PGF2 alpha) for 4-8 days. In normal eyes, collagen fibrils and microfibrils are in places in continuity with the muscle cells' basal lamina, which is connected to the cell membrane by fine fibrillous material. In old eyes, the basal lamina is markedly thickened, masking the connection of fibrils with the muscle cells' membrane. A distinctive finding in several muscle cells of old eyes are electronlucent clefts, 60-80 nm wide, between basal lamina and cell membrane, which are not transversed by fibrils or fibrillous material. The cell membrane of these muscle cells shows large folds filled with disarranged myofilaments. Additionally, these cells contain inclusion bodies consisting of concentrically arranged double membranes. Following treatment with PGF2 alpha, similar changes are seen in young animals, too. Here, the muscle cells have lost their connection to the extracellular fibrils due to a PGF2 alpha-induced lysis of extracellular material. Lack of attachment between basal lamina and altered muscle cells in aged eyes might indicate an involvement of the extracellular matrix in age-related changes of the individual ciliary muscle cells.     

Mhc-DQ-DRB haplotype analysis in the rhesus macaque: evidence for a number of different haplotypes displaying a low allelic polymorphism

In the HLA-DRB subregion of man, five major groups of haplotypes, often displaying a remarkable polymorphism, are distinguishable. The polymorphism is thought to be generated by point mutation, microgene conversion and gene rearrangement by recombination. In order to gain insight into the organization of the rhesus macaque major histocompatibility complex (MHC) class II region, DRB genes from monkeys of different origins previously typed for their DQ genes were analyzed. At first DRB haplotypes were deduced from DQ-homozygous monkeys. The stability of these haplotypes was then examined in DQ-heterozygous monkeys by sequence-based typing for the presence of members of the DRB1*03 and DRB1*04 lineage, and for seven single alleles detected on the haplotypes. Six DRB haplotypes linked to the five most frequent and three haplotypes linked to less frequent DQ haplotypes were identified. Six novel DRB alleles were detected. The number of DRB genes per haplotype varied between two and four. The results altogether suggest that in rhesus macaques, in comparison to man, the DQ haplotypes are linked to only a small number of DRB haplotypes, the number and diversity of DRB haplotypes is larger, and the allelic polymorphism of a given haplotype is smaller. The diversity of the DRB haplotypes was partly due to the varying number and identity of genes linked to DRB1*03 and DRB1*04. Furthermore, the number of DRB1 genes themselves varied from zero to two.     

Neuronal activity related to visually guided saccades in the frontal eye fields of rhesus monkeys: comparison with supplementary eye fields.

1. The purpose of this study was to analyze the response properties of neurons in the frontal eye fields (FEF) of rhesus monkeys (Macaca mulatta) and to compare and contrast the various functional classes with those recorded in the supplementary eye fields (SEF) of the same animals performing the same go/no-go visual tracking task. Three hundred ten cells recorded in FEF provided the data for this investigation. 2. Visual cells in FEF responded to the stimuli that guided the eye movements. The visual cells in FEF responded with a slightly shorter latency and were more consistent and phasic in their activation than their counterparts in SEF. The receptive fields tended to emphasize the contralateral hemifield to the same extent as those observed in SEF visual cells. 3. Preparatory set cells began to discharge after the presentation of the target and ceased firing before the saccade, after the go/no-go cue was given. These neurons comprised a smaller proportion in FEF than in SEF. In contrast to their counterparts in SEF, the preparatory set cells in FEF did not respond preferentially in relation to contralateral movements, even though most responded preferentially for movements in one particular direction. The time course of the discharge of the FEF set cells was similar to that of their SEF counterparts, except that they reached their peak level of activation sooner. The few preparatory set cells in FEF tested with both auditory and visual stimuli tended to respond preferentially to the visual targets, whereas, in contrast, most set cells in SEF were bimodal. 4. Sensory-movement cells represented the largest population of cells recorded in FEF, responding in relation to both the presentation of the targets and the execution of the saccade. Although some of these sensory-movement cells resembled their counterparts in SEF by exhibiting a sustained elevation of activity, most of the FEF sensory-movement cells gave two discrete bursts, one after the presentation of the target and another before and during the saccade. Like their counterparts in SEF, the sensory-movement cells tended to be tuned for saccades into the contralateral hemifield, but this tendency was more pronounced in FEF than in SEF. The FEF sensory-movement cells discharged more briskly, with a shorter latency relative to the presentation of the target, than their counterparts in SEF. In addition, the FEF sensory-movement neurons reached their peak activation sooner than SEF sensory-movement neurons. Most FEF sensory-movement cells exhibited different patterns of activation in response to visual and auditory targets.(ABSTRACT TRUNCATED AT 400 WORDS)     

Saline infusion contrast intrauterine sonographic assessment of the endometrium with high-frequency, real-time miniature transducer in normal menstrual cycle: a preliminary report.

Normal endometrial texture was visualized using saline infusion contrast intrauterine sonography with a specially developed 20 MHz flexible catheter-based high-resolution, real-time miniature (2.4 mm outer diameter) ultrasound transducer in primary infertile women (n = 15) with a normal menstrual cycle. All the women had <2 years infertility duration and were studied in proliferative, and early or mid-secretory phases. Before intrauterine sonography, transvaginal sonographic assessment of the endometrium was conducted. The overall image clarity was subjectively compared between intrauterine and transvaginal sonography. Most endometrial textures in both proliferative and secretory phases were viewed more easily with intrauterine rather than transvaginal sonography, and this was especially true with an intrauterine saline infusion technique. Moreover, it was possible to obtain finer image quality of very small endometrial interfacial and internal textures with intrauterine sonography. However, the depth of penetration of the ultrasound beam is only approximately 2 cm, therefore examination of larger pathological endometrial lesions is markedly limited because of the shallow scanning range of the high-frequency transducer. Intrauterine sonography may be a valuable tool in imaging endometrial texture in normal menstrual cycle, and possibly in infertility practice, complementing and not replacing transvaginal sonography.     

Distribution of uteroglobin in the rabbit endometrium after treatment with an anti-progesterone (ZK 98.734): an immunocytochemical study

The effect of the synthetic steroid ZK 98.734, an anti-progesteronewith high affinity for the progesterone receptor, on uteroglobindistribution in the rabbit endometrium has been studied by meansof immunocytochemistry. Rabbits were treated with ZK 98.734during the second, third and fourth day of pseudopregnancy.From the fifth up to the eighth day of pseudopregnancy the uteriwere processed for immunocytochemistry using the peroxidase-antiperoxidase(PAP) and protein A -gold techniques. Uteroglobin synthesisand release could be inhibited by the anti-progesterone treatment.On day 5 and 6 there was no labelling of the uterine secretionsand only a few diffusely labelled non-ciliated cells could beseen in the surface and glandular epithelium. The inhibitionwas reversible in so far as on day 7 and day 8 the rabbit endometriumexhibits a clear labelling of the uterine secretion as wellas an increase in positive reaction in the epithelial cellslining the glands. In all treated animals the intracellularuteroglobin labelling was confined to the Golgi complex andsecretory vesicles with a significant increase from the fifthto the eighth day of pseudopregnancy. Together with the describedmorphological changes these results indicate that ZK 98.734is capable of inducing a delayed secretion in the rabbit endometrium,which is comparable to the delay in secretion caused by post-coitaloestradiol treatment. However, the antigestagen effect is probablydue to a different mechanism of endocrine interference withpre-implantation. The most exciting consequence, so far, isthe prolongation of progesterone action after the anti-progesteronetreatment had ended.     

The role of pre-ovulatory progesterone in the midcycle gonadotrophin surge, ovulation and subsequent luteal phase: studies with RU486 in rhesus monkeys

Conflicting evidence exists on the possible physiological roleof progesterone in the regulation of the midcycle surge of gonadotrophinsduring the normal primate menstrual cycle. We designed the presentstudy based on the availability of a potent antiprogesterone,RU486, that acts by binding to the progesterone receptor withoutinducing progestational activity. Regularly cycling rhesus monkeysreceived daily administration of RU486,10 mg orally (n = 8)or vehicle (n = 5) from the day of the menstrual cycle in whichserum oestradiol was 130 pg/ml or more, and a laparoscopy revealedthe presence of a dominant follicle. While vehicle administrationdid not affect the normal ovulatory pattern nor the hormonalmilieu of the menstrual cycles, RU486 induced marked aberrationsduring the treated cycles. Delay of ovulation with a normalsubsequent luteal phase was observed in three animals. Threeanimals remained anovulatory until the following cycle and twoanimals that ovulated on days 14 and 16 of the treated cycleshad short luteal phases. Analysis of daily FSH, LH, oestradioland progesterone revealed that the administration of RU486 disruptedthe midcycle pattern of gonadotrophins by disrupting them afterthe surge was initiated. Oestradiol surges were not differentfrom controls and in all animals the ascendant levels of progesteronewere interrupted by the administration of the antiprogesterone.This study clearly shows that the pre-ovulatory administrationof RU486, a potent antiprogesterone, alters pre-ovulatory gonadotrophinsecretion, inducing different degrees of menstrual irregularitiessuch as anovulation, delayed ovulations and short luteal phases.Based on these results it is possible to speculate that progesteroneexerts a facilitatory effect (positive feed-back) on the midcyclegonadotrophin peak that induces ovulation in primates.     

Intravascular fasciitis of the hip joint in a postpartum female: misdiagnosed as low grade fibromyxoid sarcoma

Intravascular fasciitis is a special type of nodular fasciitis. It is an uncommon lesion. We report here the first case of intravascular fasciitis involving the deep muscle of the hip joint. The female, postpartum patient presented with a large, firm, painless tumoral mass in the anterolateral muscles of the right hip. The diagnosis of intravascular fasciitis was difficult because of the large size and location of the lesion. The false positive immunohistochemistry for MUC4 initially caused our team to misdiagnose the intravascular fasciitis as a low grade malignant fibromyxoid sarcoma. Our case adds to the literature on intravascular fasciitis. Since the operation about 20 months prior, the tumor in this case has not recurred.     

Bone mineral density during maintenance treatment with supraphysiological doses of levothyroxine in affective disorders: a longitudinal study

BACKGROUND: This prospective study was designed to determine whether patients with prophylaxis-resistant affective disorders, receiving adjunctive maintenance therapy with supraphysiological doses of levothyroxine (L-T4), show evidence of accelerated bone loss compared to the reference population database. METHODS: In 21 patients, bone mineral density (BMD) of the spine (lumbar vertebrae L1-L4) and femur (femoral neck, trochanter, and Ward's triangle) was measured by dual energy X-ray absorptiometry (DXA). BMD measurement was performed first after patients had been on thyroid-stimulating hormone (TSH)-suppressive therapy with L-T4 (mean dose=411 mcg/d) for an average of 16.4 months and again after 33.6 months of L-T4 (mean dose=416 mcg/d) therapy. RESULTS: There was no statistically significant difference between the actual percentage decline in bone mineral density and the expected percentage decline in any of the measured bone regions. In a stepwise linear regression analysis, age was identified as a predictor of percentage change in BMD. After controlling for age, the only other variable that showed a consistent trend was the dose of L-T4, with higher doses being positively correlated with the percentage decline of BMD. LIMITATIONS: Relatively small sample size, no bone density assessment prior to treatment with L-T4, no patient control group with mood disorders who did not receive L-T4 treatment, and bone density follow-up intervals were variable. CONCLUSIONS: This study did not demonstrate evidence that long-term treatment of affectively ill patients with supraphysiological doses of L-T4 significantly accelerates loss of bone mineral density compared to the age-matched reference population. However, the decline of BMD in one individual patient underscores that caution is indicated and that regular assessment of BMD during longer-term supraphysiological thyroid hormone treatment is needed.