Behavioral responses to ovariectomy and chronic anabolic steroid treatment in female cynomolgus macaques.

A pilot study was conducted to investigate the effects of ovariectomy on rates of aggressive and affiliative behavior, as well as body size, in 38 young adult female cynomolgus monkeys (Macaca fascicularis) living in isosexual social groups of four to five animals. In addition, we assessed the effects of nandrolone decanoate (an anabolic steroid used for postmenopausal hormone replacement therapy) on indices of aggression, submission, and body size. Animals were randomized into three experimental conditions: 1) sham ovariectomized, untreated (SHAM); 2) ovariectomized, untreated (OVX); and, 3) ovariectomized, treated with nandrolone decanoate for 24 months (NAN). Each individual was observed for 10 min, one to two times per month, and all instances of agonistic and affiliative behavior were recorded by means of focal animal sampling. Ovariectomized, untreated animals exhibited a two- to threefold increase in aggression compared to SHAM or NAN animals; F(2, 32) = 4.09, p = 0.026; however, the expression of prosocial or affiliative behaviors as measured by rates of grooming and initiating friendly behavior was unaffected. At an i.m. dose of 25 mg every 2 weeks, nandrolone decanoate caused a 60% increase in body weight of the animals compared to untreated intact and ovariectomized animals, F(2, 31) = 161.57, p < 0.0001.

A review and methodologic critique of the literature refuting whiplash syndrome.

The validity of whiplash syndrome has been a source of debate in the medical literature for many years. Some authors have published articles suggesting that whiplash injuries are impossible at certain collision speeds; others have stated that the problem is psychological, or is feigned as a means to obtain secondary financial gain. These articles contradict the majority of the literature, which shows that whiplash injuries and their sequelae are a highly prevalent problem that affects a significant proportion of the population. The authors of the current literature critique reviewed the biomedical and engineering literature relating to whiplash syndrome, searching for articles that refuted the validity of whiplash injuries. Twenty articles containing nine distinct statements refuting the validity of whiplash syndrome were found that fit the inclusion criteria. The methodology described in these articles was evaluated critically to determine if the authors' observations regarding the validity of whiplash syndrome were scientifically sound. The authors of the current critique found that all of the articles contained significant methodologic flaws with regard to their respective authors' statements refuting the validity of whiplash syndrome. The most frequently found flaws were inadequate study size, nonrepresentative study sample, nonrepresentative crash conditions (for crash tests), and inappropriate study design. As a result of the current literature review, it was determined that there is no epidemiologic or scientific basis in the literature for the following statements: whiplash injuries do not lead to chronic pain, rear impact collisions that do not result in vehicle damage are unlikely to cause injury, and whiplash trauma is biomechanically comparable with common movements of daily living.

The iceman under pressure (Part I): A description of skull deformations due to 5100 years of glacial action.

Due to its long deposition in the glacier, the 'Iceman' (an ice-mummy from the Hauslabjoch) has been deformed, notably its skull. We introduce various comparative methods that describe these deformations, assuming they can be ascribed--to a large extent--to glacial action. While pressure is a scalar, the deformations must be described via a 2-tensor strain field (which can be represented by a matrix function value at every point throughout the skull). In this paper, we present the assumed deformations in numerous graphical forms and, furthermore, the limitations in interpretation--including an estimate of statistical variability--that can be revealed by this analysis. These methods, although describing the results of glacial action and implying a 2-tensor strain field (which will be presented in a subsequent paper), do not permit a straightforward reconstruction of the original, underformed skull. These methods have wider applications to the general problem of deformation.

Re-evaluation of the endocranial volume of the Guattari 1 Neandertal specimen (Monte Circeo).

The endocranial capacity of Guattari 1 originally was estimated by Sergi as approximately 1.550 cm3. Using three different approaches, a physical endocast, a stereolithographic model, and a virtual endocast, we have estimated the endocranial capacity of Guattari 1 as approximately 1.350 cm3. This paper explains our revision of the estimated endocranial volume of Guattari 1, provides a cautionary case concerning other estimates of endocranial volume, and demonstrates and encourages the use of recent advances in imaging, modeling, and analysis of endocranial volume.

Nandrolone decanoate causes pathologic changes in the uterus of surgically postmenopausal female cynomolgus macaques.

OBJECTIVE: To determine the effects of the androgenic anabolic steroid nandrolone decanoate on uterine endometrium and myometrium and on the mammary gland of female cynomolgus macaques by using morphologic, histomorphometric, and histopathologic determinations. DESIGN: Histologic and histomorphometric measurements were performed on uteri and mammary glands that were collected at necropsy from animals that had been used in a long-term experiment to examine the effects of nandrolone decanoate on bone and coronary arteries. The animals were surgically postmenopausal cynomolgus macaques randomized into four treatment groups: (a) intact sham ovariectomized (sham; n = 12), (b) ovariectomized (OVX; n = 15), (c) ovariectomized + nandrolone decanoate for 2 years (OVX + ND; n = 14), and (d) ovariectomized + nandrolone decanoate for 1 year, beginning 1 year after ovariectomy (OVX + NDdelay; n = 11). Intramuscular injections of nandrolone decanoate (25 mg every 3 weeks) were given to the two nandrolone-treated groups of animals (OVX + ND and OVX + NDdelay): one starting 3 weeks after ovariectomy and continuing for 2 years and the other group 1 year after ovariectomy. The sham and OVX groups were given an intramuscular injection of sterile vehicle every 3 weeks. RESULTS: Nandrolone treatment was moderately uterotropic in all treated versus ovariectomized animals. Changes induced were an increase in uterine weight, endometrial thickness, and glandular area, and a high incidence of mucometra. Glandular architecture was altered by nandrolone treatment such that glands extended into the myometrium (producing an adenomyosis-like lesion). Mammary gland changes were mild and equivocal. CONCLUSION: Nandrolone induced pathologic changes in ovariectomized monkeys similar to adenomyosis in the uterus.

The androgenic anabolic steroid nandrolone decanoate prevents osteopenia and inhibits bone turnover in ovariectomized cynomolgus monkeys.

We examined the effects of nandrolone decanoate (25 mg im every 3 weeks) on bone mass, serum biomarkers, and bone histomorphometric endpoints in 52 female cynomolgus macaques randomized into four treatment groups: (1) sham-ovariectomized (sham); (2) ovariectomized + placebo for 2 years (ovx); (3) ovx + nandrolone decanoate for 2 years (Nan); and (4) ovx + nandrolone decanoate beginning 1 year after ovx (dNan). Serum alkaline phosphatase (ALP), osteocalcin, and tartrate-resistant acid phosphatase (TRAP) were assayed every 3 months, and X-ray densitometry of the lumbar spine was done every 6 months. Fluorochrome-labeled iliac biopsies collected at baseline and 1 year, and lumbar vertebrae and midshaft femur collected at 2 years, were evaluated histomorphometrically. Body weight increased over 50% with administration of nandrolone. After 2 years, ovx animals had lower spinal BMC and BMD than all other groups. Ovx animals also had higher bone turnover rates than all other groups, as indicated by higher levels of the serum and urine biomarkers, and by at least twofold higher label-based bone formation rates in the femur diaphysis and in both cancellous and cortical bone of the ilium and vertebral bodies. Nandrolone-treated animals had similar serum estradiol levels as the sham animals, presumably due to conversion of endogenous or exogenous androgens. The effects of nandrolone on bone in this experiment are consistent with estradiol action and may be attributable to the increased serum estradiol. Despite >50% higher body weight, nandrolone-treated, ovariectomized animals did not have higher bone mass than sham animals.

Effects of the anabolic steroid nandrolone decanoate on plasma lipids and coronary arteries of female cynomolgus macaques.

In this study, we examined the effect of nandrolone decanoate, and anabolic steroid (AS), on plasma lipid concentrations and coronary arteries of female cynomolgus monkeys fed a moderately atherogenic diet. There were four treatment groups: (1) intact, sham-ovariectomized (n = 12); (2) ovariectomized (OVX) + placebo for 2 years (n = 15); (3) OVX + nandrolone decanoate for 2 years (n = 14); and (4) OVX + nandrolone decanoate beginning 1 year after ovariectomy (n = 11). Serial blood samples were analyzed for total plasma cholesterol (TPC), high-density lipoprotein cholesterol (HDL-C), very-low-density lipoprotein (VLDL-C) plus low-density lipoprotein (LDL-C) cholesterol, and estradiol. All animals were necropsied after 2 years, and the coronary arteries were evaluated. There was no difference in plasma lipid concentrations between groups (P > .05) at any time. Coronary artery atherosclerosis extent (plaque size) was significantly greater in the group administered nandrolone for 2 years compared with the intact sham-operated group (P < .05), but not with the OVX + placebo group. The groups administered nandrolone had significantly larger arteries than the other two groups. Lumen area was significantly larger in the group given nandrolone for 1 year compared with all other groups (P < .05). All artery effects remained after controlling the statistical analysis for body weight. Longer-term treatment with nandrolone resulted in increased plaque size, and therefore, the possible benefit of increased lumen area was compromised. The data also suggest that nandrolone was converted to estradiol, and this conversion also may play a role in the arterial and lipid effects observed.

Development of osteopenia in ovariectomized cynomolgus monkeys (Macaca fascicularis).

Spinal osteopenia that is due in part to failure to gain bone has previously been reported in ovariectomized nonhuman primates. In these studies, development of osteopenia over one year was followed by dual-energy x-ray absorptiometry in both domestically-reared and feral ovariectomized (OVX) and sham-ovariectomized (SHAM) cynomolgus monkeys. To promote development of absolute osteopenia, the domestically-reared animals were all older than nine years and were fed a diet containing 0.14% calcium for most of the experimental period. Both SHAM and OVX feral animals fed 0.6% calcium gained bone mass, with significantly lower rates of gain in SHAM monkeys. OVX domestically-reared monkeys lost bone during one year, while SHAM domestically-reared animals showed no significant change from baseline. Thus, relative osteopenia developed in both experiments, but only the domestically-reared animals developed absolute osteopenia. Nonhuman primates are the only animal model shown to develop absolute osteopenia after ovariectomy. These data suggest that absolute osteopenia develops after ovariectomy in monkeys with stable pre-ovariectomy bone mass which are fed a level of calcium comparable to that consumed by American women.

Osteoarthritis in cynomolgus macaques: a primate model of naturally occurring disease.

The objective of the present study was to determine if naturally occurring osteoarthritis of the knee joints that is similar to the condition in humans develops in cynomolgus macaques. Knee joints from 58 young adult (mean age, 7.4 years) female cynomolgus macaques were studied with x-ray densitometry, high-detail radiography, and histology. The animals studied were subjects in a triad designed to examine the effects of the administration of sex steroids on atherosclerosis; except for a control group, the monkeys had been either ovariectomized or treated with sex steroids for 2 years. Therefore, the data were analyzed to determine if these treatments, both of which can influence bone density, affected the severity of osteoarthritis. There was a high prevalence of osteoarthritic lesions, morphologically similar to those seen in humans. Bone changes were more common and severe than cartilage changes and morphologically appeared to precede the cartilage changes. Treatment with testosterone resulted in increased body weight, body mass index, and bone mineral content in the femur and tibia but did not affect the severity of osteoarthritis. These data indicate that naturally occurring osteoarthritis developed in the knee joints of cynomolgus macaques; these animals may be a useful model for the study of osteoarthritis in humans.

Bone functional changes in intact, ovariectomized, and ovariectomized, hormone-supplemented adult cynomolgus monkeys (Macaca fascicularis) evaluated by serum markers and dynamic histomorphometry.

Several parameters of bone mass and function were investigated in three experiments involving intact, ovariectomized, or hormone-supplemented ovariectomized female cynomolgus monkeys. Ovariectomized animals had increased serum levels of alkaline phosphatase and acid phosphatase compared with intact and hormone-supplemented animals. Vertebral bone mass measured ex vivo by dual-photon absorptiometry was reduced by 11-19% in ovariectomized animals compared with intact and hormone-supplemented animals. The most dramatic effects observed with ovariectomy were markedly increased (30-60%) bone formation rates in vertebral cancellous bone, primarily caused by higher activation frequency of basic multicellular units of bone. In addition, combined resorption and reversal periods were decreased and formation period increased in untreated ovariectomized animals. Changes in static histomorphometry parameters were less dramatic, cancellous bone volume being 1-14% lower in ovariectomized animals compared with intact or ovariectomized hormone-supplemented animals. The data indicate that changes in bone resorption are primarily responsible for the lower bone mass of estrogen deficiency and increased bone mass in hormone-supplemented animals. Bone changes in ovariectomized cynomolgus monkeys resemble those in women after menopause and similarly respond positively to hormone supplementation. As such, cynomolgus monkeys are an excellent model for studying the basic mechanisms of osteoporosis and for the development of suitable therapeutic regimens.

Bone mass in female cynomolgus macaques: a cross-sectional and longitudinal study by age.

A cross-sectional study by age was designed to evaluate and describe the bone mineral content (BMC, g) and density (BMD, g/cm2) in a population of female cynomolgus macaques (Macaca fascicularis). Dual-energy X-ray absorptiometry was used to measure, in segments L2-L4 of the lumbar spine, the BMC (BMCs), BMD (BMDs), length, and total-body BMC(BMCTB) in 171 female monkeys ranging in age between 3.7 and 22.0 years. The animals were divided into three age groups: (1) young (< 6.5 years, n = 51); (2) adult (> 6.5 years and < 10.5 years, n = 63); and (3) mature (> 10.5 years, n = 57). Young animals had a significantly lower (P < 0.05) body weight and shorter trunk length than adult or mature animals. Young animals also had significantly less (P < 0.05) BMCS, BMDS, and BMCTB than adult or mature animals, and had significantly shorter (P < 0.01) lumbar spine vertebral segments than the other two groups. Longitudinally, 63 animals had repeated lumbar spine scans to examine changes over time. Young animals showed a positive and significant change (P < 0.05) in BMCs and BMDs through time, whereas these parameters did not change in adult animals, and mature animals had a trend towards bone loss through time. Densitometric results suggested that peak bone mass in the lumbar spine was achieved by 9 years of age. Radiographic and dental criteria were developed to identify animals that had reached peak bone mass, and the combined radiographic and dental scoring system reliably identified animals 9 years and older. Female cynomolgus macaques 9 years old or older are recommended for investigations of bone remodeling and associated conditions, such as osteoporosis.

Accuracy and precision of lumbar bone mineral content by dual-energy X-ray absorptiometry in live female monkeys.

Dual-energy X-ray absorptiometry (DXA) was used to determine the in vivo bone mineral content (BMC) of lumbar vertebrae in 20 feral adult female cynomolgus macaques (Macaca fascicularis). The ash weight of the third lumbar vertebra (L3) was compared to the measured L3BMC of the in vivo DXA analyses. Correlation between the estimated L3BMC by DXA and the actual ash weight was significant (r = 0.965, P less than 0.01); however, DXA methodology underestimated ash weight on the average of 6.2%. Correlation was significant between two sequential in vivo DXA scans (r = 0.988, P less than 0.001). Noninvasive in vivo DXA was a fast, precise, and effective method for measuring the lumbar BMC in female cynomolgus macaques.

Effects on bone of surgical menopause and estrogen therapy with or without progesterone replacement in cynomolgus monkeys.

The influence of estrogen replacement therapy on bone loss of surgically postmenopausal cynomolgus macaques was evaluated histomorphometrically using the first (L-1) lumbar vertebra and ex vivo dual photon absorptiometry of the third (L-3) lumbar vertebra. The animals were a subgroup of a larger study on the effects of estrogen replacement therapy on diet-induced coronary artery atherosclerosis. The three experimental conditions were as follows: untreated females with oophorectomy, females with oophorectomy treated with continuous estrogen replacement therapy plus cyclic progesterone, and females with oophorectomy treated with estrogen replacement therapy. Bone mineral density (grams per square centimeter) of L-3, when covaried for body mass index (body mass index, body weight/(trunk length/100)2), was significantly lower for the oophorectomy group compared with the group treated by estrogen replacement therapy plus progesterone and estrogen replacement therapy groups (p = 0.018). When covaried for body mass index, trabecular bone volume percentage of a midsagittal section of L-1 was not significantly different between groups, but the adjusted mean was greatest in the estrogen replacement therapy plus progesterone group, followed closely by the estrogen replacement therapy group, and was least in the oophorectomy group. When covaried for body mass index, trabecular plate number was significantly lower (p = 0.022) and mean trabecular plate separation was significantly higher (p = 0.033) in the oophorectomy group. Thus both estrogen replacement therapy and estrogen replacement therapy plus progesterone provided overall protection against surgical menopause--associated bone mass loss. Cynomolgus macaques are an extremely useful animal model for estrogen replacement therapy use in prevention of postmenopausal bone loss.

Accuracy and reproducibility of lumbar bone mineral status determined by dual photon absorptiometry in live male cynomolgus macaques (Macaca fascicularis).

Dual photon absorptiometry (DPA) was used to determine the in vivo bone mineral content (BMC) and bone mineral density of lumbar vertebrae (L2-4) in feral adult male cynomolgus macaques (Macaca fascicularis, n = 20). Following in vivo DPA scans, all animals were euthanized and the lumbar spine segment excised. The excised lumbar vertebrae, plus spinal cord, were formalin-fixed and measured three times by ex vivo DPA. The first ex vivo scan matched the monkey's own in vivo soft tissue beam attenuation ratio (Rs, ranging from 1.38 to 1.45). The second ex vivo scan was made creating a constant obese-like condition (Rs = 1.38), and the third creating a constant lean-like condition (Rs = 1.45). All scans were taken at a 1.0 mm point resolution, 1.0 mm line spacing, a 6 cm width, and a scan speed of 5 mm/sec. The second lumbar vertebra (L2) was ashed and the ash weight was compared to the measured L2BMC of the in vivo and ex vivo DPA analyses. Noninvasive in vivo DPA proved to be adequate in measuring the lumbar bone mineral content in male cynomolgus macaques. Ex vivo DPA at a high and constant Rs value of 1.45 provided for the best accuracy of formalin-fixed wet bone tissue when compared to the bone ash weight.

Maternal factors affecting reproduction in a breeding colony of cynomolgus macaques (Macaca fascicularis).

A breeding colony of cynomolgus macaques (Macaca fascicularis), composed of imported and colony-born animals and established for 9 years, was evaluated for maternal factors associated with reproductive failure. The factors evaluated included age, gravidity, parity, origin, previous stillbirths, clinical incidents and type of housing. The effects of each factor on pregnancy rate (PR), stillbirth rate (SR), infant mortality rate (IMR) and pregnancy success (PS) were evaluated. The overall colony rates were: PR = 53%, SR = 22%, IMR = 22%, and PS = 60%. Neonatal death rate for the group was 12%. Pregnancy rate was most affected by maternal factors. Clinical incidents occurring during pregnancy were associated with a significant increase in the stillbirth rate, but did not affect infant mortality rate. Maternal age did not affect any of the measures of reproductive output. Pregnancy rate peaked at 6-8 years of age and decreased thereafter, while pregnancy success peaked at 9-11 years of age. Gravidity and parity had a positive linear relationship with pregnancy rate.

Bone changes in oophorectomized monkeys: CT findings.

Single and dual energy CT (75 and 125 kVp) measurements of density were made on the excised lumbar vertebrae (L5) of oophorectomized (n = 17) and control (n = 19) monkeys. The single-energy results at 75 and 125 kVp indicated an annual bone diminution of 9.1 and 10.9%, respectively, but the dual-energy determination showed only a 6.6%/year loss. The difference between single- and dual-energy results could be explained by a putative increase in marrow fat of slightly greater than 100 mg/cm3. Changes in single-energy CT density with oophorectomy and aging may be substantially overestimated.

Periapical abscesses of cut canine teeth in cynomolgus macaques.

Periapical abscesses of canine teeth in a group of 47 male cynomolgus macaques are found to be associated with a procedure, involving cutting the canine teeth, used to make non-human primates less hazardous. Examination of 150 canine teeth in dry specimens revealed 30 cut teeth with no exposure of the pulpal chamber and 107 cut teeth where the dental pulp had been exposed. Of those teeth which had the pulpal chamber exposed by the cutting procedure, 85 (79.4%) showed clear evidence of osseous changes consistent with abscess formation in the apical regions.

Effects of ovariectomy on iliac trabecular bone in baboons (Papio anubis).

Biopsies were collected from the left iliac crest of six adult female baboons, after which three of the animals were ovariectomized. Biopsies were collected from the right iliac crest six months later. Histomorphometric evaluation of the biopsies revealed consistent increases in fractional forming surface, appositional rate, and volume-based bone formation rate after ovariectomy. The data indicate that bone turnover is increased following ovariectomy in the baboon.