Rewarming of the hypothermic rhesus monkey with electromagnetic radiation

To obtain more detailed information relative to the potential usefulness of using radio frequency (RF) energy in treating hypothermia, anesthetized rhesus monkeys were used in a rewarming study that compared a conventional method (heating pad) with an RF induction coil system. Rectal temperature (Tre) of each subject was monitored, and enzyme and isoenzyme levels were determined from blood samples collected before, during, and up to 48 h after hypothermia in order to assess the effects of each rewarming method. The previously observed postprocedure rise in serum enzymes (most visible at 24 h) was again seen, with no statistically significant difference in the time course of serum enzyme levels between the two treatments for comparable durations of hypothermia. To test the limits of the ability of the RF induction coil system, successively more severe hypothermia was induced in the subjects to the point of cardiovascular collapse (Tre less than 20 degrees C); RF energy was successful in resuscitating the profoundly hypothermic subjects without discernible harmful effects.     

Ultrastructural studies on the epididymal spermatozoa in the rhesus monkey

Ultrastructural studies on the spermatozoa in different regions of the epididymis of the rhesus monkey have shown that the process of sperm maturation is associated with the caudad migration of the cytoplastmic droplet, a reduction in the volume of the cytoplasmic droplet, and an obvious wrinkling of the plasma membrane surrounding the head of the spermatozoa. These changes are completed by the time the spermatozoa reach the distal-middle segment of the epididymis. The present studies also indicate that spermatozoa are incorporated into the intraepithelial cells in the epidymis. This finding suggests that spermiophagy is a normal occurrence in the epidymis of rhesus monkey.     

Far-field microwave dosimetry in a rhesus monkey model

Dosimetric measurements were made in a muscle-equivalent model of an adult rhesus monkey subjected to far-field irradiation at 1.29 GHz. Profiles of microwave-induced heating in the model were obtained at eight locations, and a gradient-layer whole-body calorimeter was used to measure total absorbed energy. Average specific absorption rate (SAR) was calculated both from the calorimeter experiments and from the local temperature measurements. Thermographic imaging techniques were used to qualitatively show the microwave-induced surface heating patterns. For this model the calculated average SAR was 0.155 (W/kg)/(mW/cm²) which, at 1.29 GHs, makes the absorption cross section 84% of the geometric shadow cross section. The SAR is about three times that predicted for a prolate spheroidal model of similar mass. A disproportionally high absorption occured in the legs of the model positioned parallel to the E-polarization because of what is believed to be partial-body resonance.     

Increase in cyclic AMP levels by relaxin in newborn rhesus monkey uterus cell culture

Summary A novel relaxin sensitive cell line of apparent smooth muscle origin has been established from a newborn rhesus monkey uterus (NRMU). NRMU cells respond to relaxin, in the presence of 1 μM forskolin, by producing intracellular adenosine 3′, 5′-cyclic monophosphate (cAMP). The increase in cAMP levels is dose, time and cell density dependent, reaching peak levels at 10 min when cells are seeded at 1×10⁵ cells/well. Specificity was demonstrated by neutralization of the relaxin activity with anti-relaxin monoclonal and polyclonal antibodies, degradation of cAMP in the presence of phosphodiesterase, and confirmation of the absence of cGMP. Three synthetic analogs of human relaxin generated a dose-related cAMP response as did synthetic native human relaxin. Natural relaxin purified from human corpora lutea tissue also generated a response similar to synthetic human relaxin. Porcine and rat relaxins also increased levels of cAMP. Insulin, but not IGF I or IGF II, was capable of increasing cAMP levels in NRMU cells, however, 200 ng/mL were required to achieve cAMP levels comparable to 6.25 ng/ml relaxin. Combinations of relaxin with insulin, IGF I or IGF II did not increase cAMP levels above levels obtained with relaxin alone. The effect on NRMU cells of other hormones, growth factors and drugs potentially present in cell culture systems or serum samples was evaluated. In combination with relaxin, oxytocin significantly decreased the cAMP production below the levels induced by relaxin alone, whereas progesterone and prostaglandin E₂ resulted in additive increases in cAMP. These data suggest that the NRMU cell line is an appropriate target tissue for studying relaxin-mediated biological responsesin vitro as well as functioning as the primary component of a relaxinin vitro bioassay. Editor's statement This paper details a smooth muscle cell line that is responsive to relaxin and provides a useful assay system for the hormone, as well as providing a model system for the study of the mechanisms of relaxin action.     

Differential preimplantation development of rhesus monkey embryos in serum-supplemented media

Several media, some augmented with amino acids, have been formulated recently, based on simplex optimization, to support the preimplantation development of mouse embryos. For the highly limited studies on preimplantation development of nonhuman primate embryos, a complex medium (CMRL-1066) has been employed. Our objective was to compare the developmental ability of rhesus monkey embryos in a simple medium containing amino acids, KSOM/AA, with the complex media used previously. Zygotes (99) were recovered following in vitro fertilization (IVF) from six monkeys, allocated to either CMRL or KSOM/AA both containing 10% fetal calf serum (FCS), and monitored daily until reaching the expanded or hatched blastocyst stage. The distribution of cells between the inner cell mass (ICM) and trophectoderm was determined at the end of culture by differential nuclear staining. Although a greater number of embryos cultured in KSOM/AA vs. CMRL developed to the morula stage (80%) and beyond (66% to expanded blastocyst), the differences were not significant. Such embryos in KSOM/AA did, however, develop at a significantly faster rate, on average, reaching the expanded blastocyst stage 26 hr earlier than did embryos cultured in CMRL. KSOM/AA embryos hatched in less time and had a higher percentage (43 vs. 34) of cells allocated to the ICM. These results indicate that a simple medium, KSOM/AA, in the presence of serum, supports the development of rhesus monkey embryos at high efficiency and at a faster rate than that observed for embryos cultured in the complex medium, CMRL-1066. © 1996 Wiley-Liss, Inc.     

Fine structure of the monkey epididymis: A correlated thin-section and freeze-cleave study

Summary The epithelium of the monkey epididymis was studied by means of freeze-fracture techniques and conventional electron microscopy. For the study of transepithelial permeability lanthanum hydroxide was used as an intercellular tracer. The epididymal epithelium consists mainly of tall columnar cells. The long stereocilia at the apical surface, similarly to microvilli, exhibit after freeze-fracture, two distinct faces: the E face, concave and with fewer membrane-associated particles, and the complementary convex P face. In the lumen unusual groups of smooth-surfaced vacuoles are present. A tight junctional network, which shows some permeability to the lanthanum tracer, is located at the apical end of the cells. Supranuclear cross-fractures clearly show the well developed Golgi cisternae and numerous vacuole profiles. The highly infolded, centrally located nucleus exhibits, after freeze-fracture, an even distribution of nuclear pores. In the perinuclear region the rough endoplasmic reticulum, which also presents pores, displays a sheet-like organization. The basal cytoplasm is filled by numerous globular profiles of membrane-bounded granules. Freeze-cleave exposes large cytoplasmic areas where the types and amount of organelles indicate an intense metabolic activity.Supported by Grant No 104.193.1.78 from PLAMIRHInvestigator of the National Council for Scientific and Technical Research (CONICET) Argentina     

Oral curcumin supplementation improves fine motor function in the middle-aged rhesus monkey

Aged individuals experience decreased fine motor function of the hand and digits, which could result, in part, from the chronic, systemic state of inflammation that occurs with aging. Recent research for treating age-related inflammation has focused on the effects of nutraceuticals that have anti-inflammatory properties. One particular dietary polyphenol, curcumin, the principal curcuminoid of the spice turmeric, has been shown to have significant anti-inflammatory effects and there is mounting evidence that curcumin may serve to reduce systemic inflammation. Therefore, it could be useful for alleviating age-related impairments in fine motor function. To test this hypothesis we assessed the efficacy of a dietary intervention with a commercially available optimized curcumin to ameliorate or delay the effects of aging on fine motor function of the hand of rhesus monkeys. We administered oral daily doses of curcumin or a control vehicle to 11 monkeys over a 14- to 18-month period in which they completed two rounds of fine motor function testing. The monkeys receiving curcumin were significantly faster at retrieving a food reward by round 2 of testing than monkeys receiving a control vehicle. Further, the monkeys receiving curcumin demonstrated a greater degree of improvement in performance on our fine motor task by round 2 of testing than monkeys receiving a control vehicle. These findings reveal that fine motor function of the hand and digits is improved in middle-aged monkeys receiving chronic daily administration of curcumin.     

Sebaceous gland adenoma in a rhesus monkey (Macaca mulatta)

A tumor mass was identified below the shoulder region of a 5-year-old male rhesus monkey (Macaca mulatta). The mass was excised and diagnosed as sebaceous gland adenoma based on the microscopic findings. Morphologically it appeared as an elevated, dome-shaped, circumscribed mass of 3.6 x 2.8 x 3.2 cm in dimension with tan speckled color. Histologically, the tumor was composed of mature, sebaceous cells (sebocytes), basal cells arranged in a mass of irregular shapes and sizes, with a characteristic appearance of poly or multilobular structure. Sebocytes were well differentiated with foamy cytoplasm in the center of the lobules and poorly or undifferentiated densely staining basal cells in the periphery of the lobules. Cellular changes in the adjacent lymph node included hyperplasia of plasma cells, macrophages and lymphoid elements with typical mitosis.     

检测恒河猴血清中trastuzumab浓度的一种直接酶联免疫竞争法

采用ELISA法建立检测恒河猴血清中trastuzumab的酶联免疫竞争法,为研究人体内trastuzumab的药物动力学学和药效学提供依据。方法的测量范围是1～100μg／mL,最低检测限为1．0μg／mL。板内精密度范围91％～107％,相对标准偏差为1．5％～4．9％。板间精密度范围102％～110％,相对标准偏差为2．7％～15．4％。方法中未显示与重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白、重组抗CD20单克隆抗体、丙种球蛋白等的交叉反应。此方法的特异性、灵敏度、精密度和准确度均满足恒河猴血清样品的分析,是检测猴和人体内trastuzumab的理想方法。     

A description of the upper thoracic autonomic nervous system in the rhesus monkey (Macaca mulatta)

The purpose of this study was to describe the autonomic innervation of the carotid sinus and heart in the rhesus monkey. Nine male rhesus monkeys (Macaca mulatta) and one male crab-eating macaque (M. fascicularis) were carefully dissected from the origin of the vagus nerves and superior cervical ganglia to the level of the fourth thoracic ganglion. The specimens were either freshly killed or obtained no later than 24 hours post mortem. The macaque monkeys were found to possess an innervation pattern that displayed features common to dog (connections between the vagus nerves and middle cervical ganglia), baboon (distinct cervical sympathetic and cervical vagal nerve trunks), and man (nerves projecting from the middle cervical and stellate ganglia to the heart). Distinct inferior cervical and first thoracic ganglia were never seen, but rather, large and well defined stellate ganglia were found. The macaque innervation pattern, when considered as a whole, most closely resembled the baboon.     

猕猴精浆纤溶酶原激活因子的来源及在精子获能中的作用

我们的前期工作表明,不育症人精液中纤溶酶原激活因子（plasminogen activator;PA）活性明显升高;给成年办和猕猴注射长效睾酮诱发无精过程中,精液PA含量也伴随上升,为进一步查明PA的来源和对精子的作用,原位杂交检测组织型PA（tPA）,尿激酶型PA（uPA）及PA抑制因子－1（PAI-1)泊mRNAs在成年健康猕附睾、前列腺和精囊中的表达。体外培养猕猴精子,培液中加入uPA、tPA及其底物纤溶酶原（plasminogen）,测试PA对精子活力、顶体反应及激活卵子的影响。结果表明,猕猴附睾、前列腺和精囊均表达tPA、uPA和PAI-1 mRNAs。加入uPA能维持精子的活力,使精子产生超激活运动,诱导顶体反应的发生,并使精子获得激活卵子的能力,这说明猕猴精浆PA除来源于睾丸外,可能主要来源于附睾及附性腺;在体外,uPA,而不是tPA,可能诱导精子获能。     

Isolation and morphologic differentiation in vitro of villous cytotrophoblast cells from rhesus monkey placenta

Summary Trophoblast is the major functional cell type of the placenta. The purpose of this study was to devise a means to isolate trophoblast cells from the monkey placenta and to examine their capacity to differentiate in vitro. Methods originally devised for the isolation of human cytotrophoblast cells produced poor yields and a low degree of purity when applied to the near-term rhesus monkey placenta. However, a procedure has been developed which allows the isolation of a cell population consisting of more than 95% cytotrophoblast based on intermediate filament immunocytochemistry. The cells sedimented between densities of 1.040 and 1.053 g/ml on continuous Percoll density gradient centrifugation. When maintained in culture the cells adhered and formed aggregates of mononuclear cells by 24 h. By 5 d in culture, immunofluorescent staining using antidesmoplakin and antinuclear antibodies revealed that most colonies consisted of large multinucleated masses similar to syncytiotrophoblast. These results demonstrate trophoblast cells from monkey placental villi can be isolated with a high degree of purity and undergo morphologic, differentiation in vitro. This preparation should enable investigators to study many functional characteristics of these cells throughout gestation. This work was supported by grants HD11658 and RR00169 from the National Institutes of Health, Bethesda, MD.     

Effect of sugar type on the survival of frozen-thawed rhesus monkey (Macaca mulatta) sperm

Sperm-freezing extenders supplemented with sugar or a combination of different sugars are widely used for the cryopreservation of nonhuman primate spermatozoa. Understanding which sugar or combination of sugars offers the highest level of cryoprotection would be beneficial for the development of sperm-freezing extenders. In the present study we aimed to investigate the effect of glucose, lactose, and raffinose separately or in combination on the cryosurvival of rhesus monkey spermatozoa. Toward that end, we prepared eight extenders by adding various types of sugars to a basic medium (BM): G-BM (0.3 M glucose), L-BM (0.3 M lactose), R-BM (0.3 M raffinose), LG-BM (0.15 M lactose+0.15 M glucose), RG-BM (0.15 M raffinose+0.15 M glucose), LR-BM (0.15 M lactose+0.15 M raffinose), and LRG-BM (0.1 M lactose+0.1 M raffinsoe+0.1 M glucose). A saline control (0.157 M sodium chloride) was also used. The results showed no significant difference in post-thaw motility when spermatozoa were frozen with G-BM, L-BM, LG-BM, RG-BM, and LRG-BM, but the post-thaw motility was significantly lower when it was frozen with R-BM, LR-BM, and the saline control. The highest plasma membrane integrity was achieved when spermatozoa were frozen with G-BM, L-BM, LG-BM, RG-BM, and LRG-BM, and the highest acrosome integrity was achieved with G-BM, L-BM, LG-BM, RG-BM, LRG-BM, and the saline control. The results indicate that the various sugars offered different protective effects. For the cryopreservation of rhesus monkey spermatozoa, glucose (monosaccharide) and lactose (disaccharide) were shown to be more suitable than raffinose (trisaccharide) for preserving spermatozoal motility, plasma membrane, and acrosome. Specifically, raffinose was detrimental to sperm acrosome integrity.     

Anesthetic management for instrumentation of the pregnant rhesus monkey

The anesthetic procedures used and the responses observed during maternal instrumentation on 38 pregnant rhesus monkeys (Macaca mulatta) during the second half of gestation are reported. A laparotomy with maternal instrumentation was performed in all animals. Anesthesia was induced with ketamine and maintained with halothane. Two animals delivered within five days of anesthesia and surgery and were unable to undergo experimentation. The remaining 36 animals underwent successful experimentation. Dysrhythmias, hypotension, and hypothermia were identified as complications of anesthesia.     

Placental transfer of synthetic and endogenous estrogen conjugates in the rhesus monkey (Macaca mulatta)

The placental transfer of conjugated and nonconjugated estrogens was compared in the pregnant rhesus monkey. Placement of catheters in the maternal and fetal circulation allowed for the sampling of blood after the administration of radiolabeled naturally occurring and synthetic estrogens to mother or fetus. In all cases, nonconjugated-estrogen placental transfer was greater than conjugated-estrogen transfer. Comparison of the conjugated estrogens suggested that diethylstilbestrol-monoglucuronide (DESG) was transferred less efficiently than the estrone-sulfate (E, S). High-performance liquid chromatog-raphy (HPLC) of selected plasma samples revealed that 50–90% of the E, S observed in the maternal circulation was cleaved after fetal administration. In contrast, HPLC of maternal or fetal plasma samples after DESG administration revealed only intact DESG. These results emphasize differences in the placental transfer of the synthetic and naturally occurring estrogen hormones.     

Effects of age,lactation, and repeated cycles on rhesus monkey copulatory intervals

Young, sexually mature female rhesus monkeys copulate on more days prior to conception than do older females, and this prolonged discrete mating period is associated with an earlier rise in serum estradiol prior to the first ovulation of the breeding season. The influence of repeated ovulatory cycles and the presence of a suckling infant on the copulatory patterns were examined in two separate analyses. Extending previous work, young, nulliparous females copulated on more days at the first ovulation of the breeding season than did older, multiparous females. However, the duration of the copulatory period at the second ovulation of the breeding season was similar and significantly shorter for both age groups. Furthermore, the presence of a suckling infant did not influence the duration of the mating periods in adult, multiparous females. The onset of copulatory behavior for all females was associated with serum estradiol concentrations of approximately 90 pg/ml, indicating that the age and cycle differences in the duration of the copulatory periods are due to the time course of serum estradiol prior to ovulation. A separate, longitudinal analysis of the duration of the mating period associated with the first ovulation of three successive breeding seasons indicated that females copulated on more days during their first ovulatory cycle of their first breeding season. These data indicate that the copulatory interval is longer for females during the first ovulation of the breeding season, and this pattern is accentuated in young, sexually mature animals.     

Laboratory assessment of temperament and environmental enrichment in rhesus monkey infants (Macaca mulatta)

This study investigated the combined effects of early temperamental characteristics and environmental enrichment on a variety of developmental measures in nursery-reared rhesus monkey infants. Twenty-three infants, reared in either standard laboratory cages or enriched environments, were tested during the 1st month of life for interactive, motor, and temperamental capabilities and characteristics. At 8 months of age, all subjects were assessed on a second series of tests designed to measure their problem-solving skills, motor capabilities, and temperamental responses under challenge. Results indicated that enrichment was associated with higher scores on subsequent problem-solving and motor tests. However, such effects were found to combine with early temperament ratings. Specifically, individuals performing best on the 8-month tests had not only been reared in enriched environments, but also had been rated low on fearfulness during the early assessment. In addition, individuals scoring poorest had been rated as fearful initially in addition to being reared without enrichment. Results indicated that while high ratings on early laboratory assessments of fearfulness may be predictive of poorer problem-solving performance under challenging conditions, these adverse effects may be partially attenuated by environmental enrichment.     

Light microscopic features of the rete testis,the vas efferens,the epididymis and the vas deferens in the adult rhesus monkey

The present study was carried out to determine the detailed histological and cytological features of the excurrent ducts of the male reproductive system in the rhesus monkey. The excurrent ducts show a regional difference in their histological features. The use of some of these features as histological markers and their possible functional significance are discussed. The epithelial cells in the different components of the excurrent duct system possess cytological features which suggest their involvement in absorption and the secretion of different products into the lumen.     

Fas, FasL, Bcl-2, and Bax in the endometrium of rhesus monkey during the menstrual cycle

To study possible role and regulation of apoptosis occurred in primate endometrium, the expression of apoptosis-related molecules, Fas, FasL, B cell lymphoma/leukaemia-2 (Bcl-2), and Bax were analyzed in relation to occurrence of apoptosis and proliferation in the cycling endometrium of the rhesus monkey using immunohistochemistry and Western blot. The cell apoptosis and proliferation were evaluated by means of in situ 3'-end labeling and Ki67 immunostaining, respectively. The results showed that the expressions of Fas, Fas ligand (FasL), Bcl-2, and Bax were co-localized predominantly in the epithelial cells of the endometrium. Modest Fas staining with no obvious change was detected throughout the menstrual cycle, while the levels of FasL and Bax protein in the epithelial cells increased in the secretory phase when apoptosis was most prevalent. In contrast, epithelial immunostaining for Bcl-2 was maximal during the proliferative phase and decreased in the secretory phase. Bcl-2 immunoreactivity was also detected in some immunocytes. The coordinated expression of Fas, FasL, Bcl-2, and Bax in the cycling endometrium of the rhesus monkey suggests that the cyclic changes in endometrial growth and regression may be regulated by the balance of these factors under the action of ovary steroids.