Skeletal Variations Research Articles

The effects of feed restriction during organogenesis on embryo-fetal development in the rat

Given the role of nutrition and body weight gain in normal development, pharmaceuticals intended to reduce appetite and promote weight loss will generate safety data that may be challenging to interpret. To aid with this, the effects of feed restriction and subsequent body weight reductions on embryo-fetal development were investigated in the rat. Groups of 20 timed pregnant female Sprague-Dawley rats were offered Certified Rodent Diet 5002 either ad libitum or in restricted amounts of 20, 15, 10, and 7.5 g/day from Gestation Day (GD) 6-17. Clinical signs, body weights, and food consumption were recorded. Cesarean sections were performed on GD 21 and fetuses were sexed, weighed, and examined for external, visceral, and skeletal development. Mean maternal body weights at the end of the feed restriction period, GD 18, were reduced 0.87 x, 0.80 x, 0.69 x, and 0.63 x control mean in the 20, 15, 10, and 7.5 g/day groups, respectively. Mean body weight gains for the restriction period inclusive, GD 6-18, were 0.49 x and 0.24 x control at 10 and 7.5 g/day, respectively, and a mean body weight loss occurred at 10 and 7.5 g/day (0.95 x and 0.85 x mean GD 6 body weight, respectively). Fetal body weights were reduced 0.95 x, 0.93 x, 0.90 x, and 0.76 x control at 20, 15, 10, and 7.5 g/day, respectively. This resulted in a reduction in gravid uterine weight at 10 and 7.5 g/day. There were no external, visceral, or skeletal malformations attributed to feed restriction. There was an increase in the skeletal variation of wavy ribs and a decrease in ossification at 7.5 g/day. These data demonstrate that feed restriction-induced reductions in maternal gestational body weight gain of approximately 50% compared to ab lib fed rats only caused a reduction in fetal body weight. Even up to a 15% maternal gestational body weight loss had no effect on embryo viability in rats, but retarded fetal growth significantly enough to induce minor changes in skeletal development. There were no external, visceral, or skeletal malformations associated with any of the levels of maternal body weight reduction or loss.

Evaluation of the Developmental Toxicity of 8-2 Telomer B Alcohol

The potential maternal and developmental toxicity of 8-2 Telomer B Alcohol was assessed in rats. Groups of 22 time-mated female Crl:CD (SD)IGS BR rats were administered oral gavage doses as suspensions of 8-2 Telomer B Alcohol in aqueous 0.5% methylcellulose from day 6 through 20 of gestation (G) at daily doses of either 0, 50, 200, or 500 mg/kg. Under the conditions of this study, adverse maternal toxicity was produced at 500 mg kg− 1 day− 1 and consisted of maternal mortality, decreased body weights and body weight gains, and increased clinical observations of toxicity. One litter at 500 mg kg− 1 day− 1 consisted of one early resorption and was believed to be secondary to overt maternal toxicity, although single conceptus litters occur historically in this strain of rats. Developmental toxicity at 500 mg kg− 1 day− 1 consisted of increased fetal skeletal variations (delayed pelvic bone ossification and wavy ribs). At 200 and 500 mg kg− 1 day− 1, there were transient reductions in maternal feed consumption. In addition, there were slight increases in the incidence of delayed fetal skull bone ossification at 200 and 500 mg kg− 1 day− 1. The no-observed-adverse-effect level (NOAEL), defined as the highest dose at which adverse effects attributable to the test substance were not detected, for both maternal and developmental toxicity, is considered to be 200 mg kg− 1 day− 1. Thus, 8-2 Telomer B Alcohol is not considered to be a selective developmental toxicant in rats. The transient and quantitative nature of the observations in the 200 mg/kg group supports the conclusion that these findings were not adverse.

Evaluation of developmental toxicity of 1-butanol given to rats in drinking water throughout pregnancy

The objective of this study was to evaluate the developmental toxicity of 1-butanol in rats. Pregnant rats were given drinking water containing 1-butanol at 0.2%, 1.0% or 5.0% (316, 1454 or 5654mg/kg/day) on days 0–20 of pregnancy. A significant decrease in maternal body weight gain accompanied by reduced food and water consumption was found at 5.0%. No significant increase in the incidence of pre- and postimplantation embryonic loss was observed in any groups treated with 1-butanol. Fetal weight was significantly lowered at 5.0%. Although a significant increase in the incidence of fetuses with skeletal variations and decreased degree of ossification was found at 5.0%, no increase in the incidence of fetuses with external, skeletal and internal abnormalities was detected in any groups treated with 1-butanol. The data demonstrate that 1-butanol is developmental toxic only at maternal toxic doses. No evidence for teratogenicity of 1-butanol was noted in rats. Based on the significant decreases in maternal body weight gain and fetal weight, it is concluded that the no observed adverse effect levels (NOAELs) of 1-butanol for both dams and fetuses are 1.0% (1454mg/kg/day) in rats.

A reconnaissance of skeletal crystallography in rhombiferans, diploporans, and paracrinoids

We characterized the skeletal crystallography of representatives of nine rhombiferan, three diploporan, and three paracrinoid species. Crystallographic data from these groups are similar to data from previous studies of crinoids, echinoids, and blastoids in that 1) orientations ofcaxes are consistent within species and within higher taxonomic groups; 2)caxes typically are oriented subparallel to the medial plane of their respective plates; and 3) the inclination of axes within the medial plane varies between taxa. Rhombiferancaxes are oriented normal to plate surfaces whereas diploporancaxes are tangential to plate surfaces. Paracrinoids more closely resemble diploporans in having irregular patterns of thecal plating but theircaxes are approximately perpendicular to plate surfaces as in rhombiferans.In contrast tocaxes,aaxes in all specimens show little regularity and cannot be distinguished from random orientations.The rhombiferanCaryocrinites ornatusdisplays minor differences in the inclinations ofcaxes depending on the location of skeletal elements on the theca. Plates at the base of the theca have slightly aborally inclined axes, whereas distal plates have axes inclined slightly adorally. This pattern matches orientations in some early crinoids, suggesting similarities between rhombiferans and crinoids in development or skeletal construction.Skeletal crystallography in various echinoderms can be compared in light of hypotheses of homology proposed in the Extraxial-Axial Theory (EAT). Skeletal elements homologized under the EAT do not correspond to any particular crystallographic axis orientation, suggesting that the homologies proposed in the EAT encompass significant underlying skeletal variation.

A RECONNAISSANCE OF SKELETAL CRYSTALLOGRAPHY IN RHOMBIFERANS, DIPLOPORANS, AND PARACRINOIDS

We characterized the skeletal crystallography of representatives of nine rhombiferan, three diploporan, and three paracrinoid species. Crystallographic data from these groups are similar to data from previous studies of crinoids, echinoids, and blastoids in that 1) orientations of c axes are consistent within species and within higher taxonomic groups; 2) c axes typically are oriented subparallel to the medial plane of their respective plates; and 3) the inclination of axes within the medial plane varies between taxa. Rhombiferan c axes are oriented normal to plate surfaces whereas diploporan c axes are tangential to plate surfaces. Paracrinoids more closely resemble diploporans in having irregular patterns of thecal plating but their c axes are approximately perpendicular to plate surfaces as in rhombiferans. In contrast to c axes, a axes in all specimens show little regularity and cannot be distinguished from random orientations. The rhombiferan Caryocrinites ornatus displays minor differences in the inclinations of c axes depending on the location of skeletal elements on the theca. Plates at the base of the theca have slightly aborally inclined axes, whereas distal plates have axes inclined slightly adorally. This pattern matches orientations in some early crinoids, suggesting similarities between rhombiferans and crinoids in development or skeletal construction. Skeletal crystallography in various echinoderms can be compared in light of hypotheses of homology proposed in the Extraxial-Axial Theory (EAT). Skeletal elements homologized under the EAT do not correspond to any particular crystallographic axis orientation, suggesting that the homologies proposed in the EAT encompass significant underlying skeletal variation.

Developmental toxicity evaluation of ibuprofen and tolmetin administered in triple daily doses to Wistar CRL:(WI)WUBR rats

Ibuprofen and tolmetin are popular non-steroidal anti-inflammatory drugs. Previous animal studies taken with single daily doses showed their good prenatal tolerability. However, since both cyclooxygenase (COX) inhibitors have a short half-life, the current report presents drug developmental effects after triple daily doses administration, as they are used in human. Drugs were separately, orally dosed to pregnant rats triple daily 8 hr apart from day 8 to 21 (GD=1-plug day). The total daily doses were set at 25.5, 255.0, and 600.0 mg/kg for ibuprofen and 25.5, 255.0, and 2550.0 mg/kg for tolmetin. Fetuses were delivered on GD 21 and routinely examined. Comprehensive clinical and developmental measurements were done. Maternal toxicity and intrauterine growth retardation were found in groups exposed to the highest doses of both drugs. An increase of external variations was reported in groups exposed to the middle and highest dose of ibuprofen and to the highest dose of tolmetin. Skeletal variations were significantly different only in litters treated with the highest doses of the drugs. Pooled statistical analysis showed a higher incidence of midline and ventricular septal (VSD) defect in rat fetuses exposed to COX inhibitors when compared with historical control data. For ibuprofen, the influence on VSD was similar to aspirin. Both COX inhibitors were toxic to dams in the highest doses evaluated, which caused a significantly greater incidence of intrauterine growth retardation and developmental variations.

Skeletal and dental variation within and between Mystacina species in southern New Zealand

Dental and skeletal measurements of the lesser short‐tailed bat Mystacina tuberculata from Codfish Island were compared with those from the greater short‐tailed bat M. robusta from the southwest Muttonbird (Titi) Islands off Stewart Island. All M. robusta were larger than any M. tuberculata and differed in shape of teeth, as measured by lengths and widths, and also had relatively smaller wing elements given other size differences. The population of M. tuberculata on Codfish Island comprises significantly larger individuals, as indicated by maxillary teeth measurements, than the late Holocene population of M. tuberculata in northern South Island. Such a latitudinal trend is the opposite of that known for M. robusta. The differences in adjacent populations and the different morphological response of the two taxa with latitude strongly support the distinction of M. robusta and M. tuberculata at a species level. These data suggest that skeletal data have more discriminatory power than external measurement data for assessing interpopulation or intertaxon variation in short‐tailed bats.

Axial skeletal and Hox expression domain alterations induced by retinoic acid, valproic acid, and bromoxynil during murine development.

Retinoic acid (RA) alters the developmental fate of the axial skeletal anlagen. "Anteriorizations" or "posteriorizations," the assumption of characteristics of embryonic areas normally anterior or posterior to the affected tissues, are correlated with altered embryonal expression domains of Hox genes after in utero RA treatment. These "homeotic" changes have been hypothesized to result from alterations of a "Hox cod" which imparts positional identity in the axial skeleton. To investigate whether such developmental alterations were specific to RA, or were a more general response to xenobiotic exposure, CD-1 pregnant mice were exposed to RA, valproic acid (VA), or bromoxynil (Br) during organogenesis. Additionally, the expression domains of two Hox genes, Hoxa7 and Hoxa10, were examined in gestation day (GD) 12.5 embryos obtained from control, RA, VA, or Br, treated gravid dams exposed on GD 6, 7, or 8. The anterior expression boundary of Hoxa7 is at the level of the C7/T1 vertebrae and that of Hoxa10 is at L6/S1. Compound-induced changes in the incidence of skeletal variants were observed. These included supernumerary cervical ribs (CSNR) lateral to C7, 8 vertebrosternal ribs, supernumerary lumbar ribs (LSNR) lateral to L1, extra presacral vertebrae, and the induction of vertebral and/or rib malformations. RA and VA administration on GD 6 caused posteriorization in the cervico-thoracic region (CSNR) while GD 8 exposure to any of the three compounds resulted in anteriorizations in the thoraco-lumbar area (LSNR and an increase in the number of presacral vertebrae). These effects occurred across regions of the axial skeleton. Analysis of gene expression demonstrated changes in the anterior boundaries of Hoxa7 expression domains in embryos treated on GD 6 and 8 with RA. VA and Br did not induce any statistically significant alterations in Hoxa7 and none of the compounds caused alterations in Hoxa10 expression domains. The studies indicate that RA GD 6 treatment-induced Hoxa7 shifts were rostral (posteriorization) while the RA-induced GD 8 anterior expression boundary shift was caudal (anteriorization), correlating with the axial skeletal changes noted. These data suggest that xenobiotic compounds such as VA and Br may induce similar axial skeletal changes by affecting different components of the developmental processes involved in the patterning of the axial skeleton.

Evaluation of developmental toxicity of β-thujaplicin (hinokitiol) following oral administration during organogenesis in rats

The objective of this study was to evaluate the developmental toxicity of β-thujaplicin (TP) in rats. Pregnant rats were given TP by gastric intubation at 15, 45, or 135 mg/kg on days 6–15 of pregnancy. The maternal body weight gain during administration at 45 and 135 mg/kg and after administration at 136 mg/kg and adjusted weight gain at 45 and 135 mg/kg were significantly reduced. A significant decrease in food consumption during and after administration was found at 45 and 135 mg/kg. A significant increase in the incidence of postimplantation loss was found in pregnant rats given TP at 135 mg/kg. A significantly lower weight was found in female fetuses at 45 and 135 mg/kg and in male fetuses at 135 mg/kg. Although a significantly increased incidence of fetuses with skeletal variations and decreased degree of ossification were found at 135 mg/kg, no significant increase in external, skeletal and internal malformations was detected after administration of TP. The data demonstrated that TP had adverse effects on embryonic/fetal survival and growth only at maternal toxic doses. No adverse effects on morphological development were found in rats fetuses. Based on the significant decreases in maternal body weight gain and weight of female fetuses at 45 mg/kg and higher, it is concluded that the no-observed-adverse-effect levels (NOAELs) of TP for both dams and fetuses are considered to be 15 mg/kg in rats.

Developmental toxicology studies of WHI-07, a novel nucleoside analogue-based dual-function microbicide, administered intravaginally to rabbits.

The zidovudine derivative, 5-bromo-6-methoxy-5,6-dihydro-3-azidothymidine-5'-(p-bromophenyl) methoxy alaninyl phosphate (WHI-07), is a dual-function spermicidal and anti-HIV agent with contraceptive and microbicidal activity. In previous subchronic and reproductive toxicity studies and a two-year carcinogenicity study, daily intravaginal application of 0.5 to 2.0% WHI-07 via a gel-microemulsion, was shown to cause no local, systemic and reproductive toxicity or increased carcinogenicity in mice. To evaluate the developmental toxicity potential of WHI-07 in a nonrodent model, subgroups of 20 superovulated NZW rabbits were artificially inseminated and exposed intravaginally to a gel-microemulsion containing 0, 0.5, 1.0, or 2.0% WHI-07 during major organogenesis [gestation days 6-18]. The dose of WHI-07 was equivalent to 1.4x10(6) to 5.7x10(6) times its anti-HIV IC50 and 1400 to 5700 times its spermicidal EC50. Throughout the duration of the experiment (GD 0-29), clinical observations, food consumption, and body weights were recorded. Reproductive and fetal parameters were evaluated following uterotomies on GD 29. Measurements included numbers of corpora lutea, pregnancy, number and distribution of implantations, resorptions, live and dead fetuses, fetal weight, sex ratio, and gross external and skeletal malformations and variations. Maternal food consumption and body weight gain were unaffected by WHI-07 treatment. Hematologic and clinical chemistry determinations on GD 19 and 29 revealed no treatment-related maternal effects. Prior studies of repeated intravaginal administration of WHI-07 gel-microemulsion revealed lack of local toxicity to rabbit vaginal mucosa. In the current study, no drug-related gross lesions were apparent at necropsy. Reproductive indices, ie, pregnancy rate, gravid uterine weights, litter size, number of corpora lutea, implantation sites, pre- and postimplantation losses, viable fetuses, resorptions, fetal body weights, and fetal sex ratio, were not affected by intravaginal exposure to WHI-07. External, and skeletal examinations of fetuses for malformations and variations did not reveal any evidence of teratogenicity in any WHI-07-treated groups. Intravaginal administration of WHI-07 at concentrations as high as 2% did not produce teratogenicity or other developmental toxicity in rabbit conceptus. These findings indicated that WHI-07 shows unique clinical potential to become the active ingredient of a new female-controlled topical microbicidal vaginal contraceptive for women who are at high risk of acquiring HIV/AIDS.

Developmental and reproductive toxicity evaluation of toluene vapor in the rat: I. Reproductive toxicity

The reproductive toxicity of toluene was evaluated in a 2-generation test in which male and female Sprague–Dawley rats, parental (F0) and first generation (F1), were exposed to toluene via whole body inhalation, 6 h/day, 7 days/week for 80 days premating and 15 days of mating at concentrations of 0, 100, 500 and 2000 ppm (0, 375, 1875 and 7500 mg/m 3). Toluene was administered at 2000 ppm to both sexes, or to females or males only to be mated with untreated partners. Pregnant females at all dose levels were exposed from gestation day (GD) 1–20 and lactation day (LD) 5–21. At LD5, females were removed from their litters for daily exposure and returned when 6 h of exposure was completed. F1 pups selected to produce the F2 generation were treated for 80 days beginning immediately after weaning (LD21) and initially mated at a minimum of 100 days of age. F2 pups were not exposed to toluene by inhalation. Toluene exposure did not induce adverse effects on fertility, reproductive performance, or maternal/pup behaviors during the lactation period in males and females of the parental or first generation, but did inhibit growth in F1 and F2 offspring in the 2000 ppm (both sexes treated) and 2000 ppm (females only treated) groups. Caesarean section of selected 2000 ppm (both sexes treated) dams at GD20 showed reduced fetal body weight and skeletal variations. Exposure to toluene caused decreased pup weights throughout lactation in F1 and F2 2000 ppm (both sexes treated), and 2000 ppm (females only treated) groups. Exposure at 2000 ppm to male parents only did not induce similar weight inhibition in offspring. The toluene offspring NOAEL is 500 ppm in groups in which maternal animals were exposed, and 2000 ppm for male only treated groups.

Skeletal Variation among Mesolithic People of the Ganga Plains: New Evidence of Habitual Activity and Adaptation to Climate

Rethinking new perspectives in South Asian archaeology necessitates wider appreciation for insights derived from the bioarchaeological analysis of prehistoric human skeletons. Since the 1970s, Mesolithic sites near Allahabad (Uttar Pradesh) have yielded abundant well-preserved human skeletons permitting a bioarchaeological approach to past life-ways. Prior research on human remains from Sarai Nahar Rai and Mahadaha is supplemented by this analysis of human skeletal variation in 47 specimens from Damdama. This report examines skeletal variation in muscle attachment sites (entheses) and musculoskeletal stress markers, prevalence of osteoarthritis, long bone dimensions and proportions, and estimates of stature for the human skeletons from Damdama. The objective of this study is to better understand habitual activity patterns, variation in stature, and adaptation to climate among Mesolithic foragers of North India. Standardized methods of paleopathology, osteometry, and stature estimation were used. While most entheses displayed a ''normal'' range of development, those associated with bipedal locomotion and overhand throwing were especially well developed. Extreme hypertrophy of the soleal line indicates repetitious and forceful plantar flexion as in walking long distances, up hills, or with heavy loads. Hypertrophy of the supinator crest suggests forceful overhand throwing as in launching spears or projectiles. Osteoarthritis is unusually low in frequency, though spinal osteophytes and arthritis of the hand and elbow were observed. Stature is tall at Damdama, a trait shared with inhabitants of Sarai Nahar Rai and Mahadaha. Collectively, North Indian Mesolithic groups are significantly taller than Eastern or Western European Mesolithic samples. Long lower limbs may be an adaptation to locomotor efficiency, but may also reflect adaptation to high seasonal temperatures. Indices of distal to proximal limb segments for both upper and lower extremities conform to physiological principles of thermoregulation and suggest biological adaptation to a hot arid environment.

Clinical outcomes of Fränkel appliance therapy assessed with a counterpart analysis

To evaluate whether the Fränkel Regulator-II (FR-II) induced mandibular growth rotations relative to the nasomaxilla and the middle cranial fossae, cephalometric changes in 28 treated Brazilian children were compared with changes in 28 untreated Class II children and in 28 children with normal occlusion. According to Enlow's counterpart analysis, the 3 groups were not significantly different initially in ramus alignment or relative ramus vertical dimension. These jaw relationships were maintained in both untreated groups. In the treated group, all children had overjet reduction, with correction of the dental arch relationship in 26 of the 28, and there was a significant trend toward a more forward ramus alignment (P =.002) and increased ramus relative vertical dimension (P =.0002). These treatment-induced changes showed a negative correlation with ramus alignment; ie, greater improvement was more likely in children who had backward ramus alignment before treatment and whose Class II malocclusion had not already been intrinsically compensated. Changes in the treated children were similar to but greater than those in the normal children, and different from those in the untreated Class II group. The data suggest that studies of skeletal variations with counterpart analysis can show ramus remodeling compensations from treatment that are missed with conventional cephalometrics. (Am J Orthod Dentofacial Orthop 2003;123:379-87)

The iceman: discovery and imaging.

The anatomic features of a 5,300-year-old mummy, the iceman, were documented with conventional radiographic, portable computed radiographic, and conventional and spiral computed tomographic images obtained between September 1991 and June 2001. A team of scientists and radiologists from Austria, Italy, and the United States supervised the examinations and interpreted the images. The images demonstrated excellent preservation of the mineralized skeleton with profound dehydration of the soft tissues. The skeleton exhibited several types of trauma, including (a) healed rib fractures, (b) hairline skull fractures and a compression deformity of the thorax, probably acquired while encased in the glacier, and (c) damage acquired during the effort to recover the corpse. Skeletal variants were present, as was evidence of degenerative arthritis, frostbite, vascular calcification, and adaptation to cultural and geographic influences. In terms of anatomy and apparent health-related conditions, the iceman was very similar to modern humans. An arrowhead lodged between the rib cage and the left scapula was the probable cause of the iceman's death. Study of the images also provided insight regarding postmortem processes that led to the iceman's mummification.

Developmental toxicities of ethylbenzene, ortho-, meta-, para-xylene and technical xylene in rats following inhalation exposure

The developmental toxicities of ethylbenzene, o-, m-, p-xylene and technical xylene were studied in Sprague–Dawley rats after inhalation exposure. Animals were exposed to either of these agents at 100, 500, 1000 or 2000 ppm, for 6 h/day, during days 6–20 of gestation. All the agents tested caused maternal toxicity expressed as a reduction in maternal body weight gain at 1000 and 2000 ppm. Decreased corrected weight gain and food consumption were observed at 1000 and 2000 ppm ethylbenzene, o-, m- or p-xylene, and at 2000 ppm technical xylene. No evidence of teratogenic effects was found after exposure to any of these agents up to 2000 ppm. Fetal toxicity evidenced by significant decreases in fetal body weights occurred at concentrations of 500 ppm or greater of o-xylene or technical xylene, and 1000 ppm or greater of ethylbenzene, m- or p-xylene. A significant increase in the mean percentage of fetuses per litter with skeletal variations was also noted at 2000 ppm ethylbenzene, o- and p-xylene. In summary, all tested agents produced developmental toxicity at 1000 and 2000 ppm, concentrations that also produced significant maternal toxicity. With o-xylene and technical xylene, developmental toxicity also occurred at 500 ppm, in the absence of maternal toxic effects. However, the only indication of a treatment-related effect was a slight decrease in fetal weight.

Clinically significant skeletal variations of the shoulder and the wrist: role of MR imaging.

Several skeletal variations of the upper extremity may cause painful conditions or precipitate early degenerative changes, either spontaneously or in response to overuse and trauma. Magnetic resonance imaging has proved particularly useful for accurate interpretation of many of these clinically significant skeletal variations; however, the widespread use of MR imaging may have contributed to over-emphasizing their clinical importance, which is still controversial in many cases. We review, illustrate, and discuss clinically significant skeletal variations of the upper extremity as seen on MR images, particularly those involving the shoulder and the wrist. In the shoulder region, we evaluate variations of acromial and coracoid processes as well as variations and minor dysplastic deformities of the glenoid fossa. We also review different skeletal variations of the carpal region, including ulnar variance, ulnar styloid, lunate morphology, carpal coalition, and carpal accessory ossicles. The role of MR imaging in assessing the clinical importance of such conditions, whether potential, controversial, or well established, is emphasized in this review.

Genetic basis for systems of skeletal quantitative traits: principal component analysis of the canid skeleton.

Evolution of mammalian skeletal structure can be rapid and the changes profound, as illustrated by the morphological diversity of the domestic dog. Here we use principal component analysis of skeletal variation in a population of Portuguese Water Dogs to reveal systems of traits defining skeletal structures. This analysis classifies phenotypic variation into independent components that can be used to dissect genetic networks regulating complex biological systems. We show that unlinked quantitative trait loci associated with these principal components individually promote both correlations within structures (e.g., within the skull or among the limb bones) and inverse correlations between structures (e.g., skull vs. limb bones). These quantitative trait loci are consistent with regulatory genes that inhibit growth of some bones while enhancing growth of others. These systems of traits could explain the skeletal differences between divergent breeds such as Greyhounds and Pit Bulls, and even some of the skeletal transformations that characterize the evolution of hominids.

Studies of 50 Hz circularly polarized magnetic fields of up to 350 μT on reproduction and embryo‐fetal development in rats: Exposure during organogenesis or during preimplantation

Groups of mated female Sprague-Dawley rats were simultaneously exposed to 0 (sham exposed), 7, 70, or 350 microT (rms) circularly polarized 50 Hz magnetic fields (MF) for 22 h/day on gestational day 8-15, the period of rat fetal organogenesis (organogenesis study) or from day 0 to day 7 of gestation, the rat preimplantation period (preimplantation study). Developmental toxicity was assessed on gestational day 20. Identical experiments were repeated to confirm reproducibility of both studies. In both studies, statistically significant differences between exposed and sham exposed animals were observed in several measured parameters; however, these differences only appeared in one, but not both replicate experiments and generally at only an isolated exposure level. Because these differences were not reproducible and did not show a dose response relationship, they were not considered related to MF exposure. In the organogenesis study, lower kidney weights of dams were seen at 70 and 350 microT in Experiment 1. Lower dam liver weights and lower mean body weights of viable female and male fetuses were seen at 70 microT in Experiment 2. Otherwise, there were no differences in these parameters or in group means for fetal loss after implantation, number of viable fetuses, fetal body weight and sex ratio, incidences of external, visceral, and skeletal abnormalities or variations, or tissue abnormalities after histopathological examination. In the preimplantation study, dam health and indices for reproduction and embryo-fetal development, including pre or postimplantation loss, number and body weight of live fetuses, and sex ratio, external, skeletal abnormalities and variations, and skeletal ossification did not differ. Dam inorganic phosphorous concentration at 350 microT was elevated in one experiment and depressed in another. In one experiment, visceral abnormalities, primarily thymic remnant in neck and accessory liver lobe, were increased in the 7 microT group. Based on these results from two studies, we conclude that circularly polarized 50 Hz MF exposure of up to 350 microT during the fetal organogenesis or during the preimplantation period does not affect reproduction and embryo-fetal development in Sprague-Dawley rats.

AN ORAL (GAVAGE) CONTROL EMBRYO-FETAL DEVELOPMENT STUDY IN THE WISTAR HANNOVER RAT

Many of the studies conducted to examine the developmental and reproductive toxicity potential of candidate pharmaceuticals use the Sprague-Dawley rat as the animal model. This is due in part to the large database for this outbred rat available for comparison of litter data, and the low incidence of fetal malformations and variations. The following study was conducted to generate information on potential embryo-fetal developmental defects and litter data in another outbred stock of rat, the Wistar Hannover. One hundred fifty pregnant female Wistar Hannover rats (Tac : Glx : WIfBR) were dosed orally once per day with distilled water from Gestation Days (GD) 6 through 17 covering the time from implantation to closure of the hard palate (GD0 = day of insemination). Caesarean sections were performed on Day 20 of gestation. All fetuses were examined for external, visceral and skeletal malformations and variations. Macroscopic and histomorphologic examinations were also completed for the F0 females at termination. The percent pregnant (88%) and litter size (average 10.6) were found to be lower than that commonly reported for the Sprague-Dawley rat (Crl : CD® (SD)BR; 95.4% and 14.6, respectively). Pre-implantation loss (14.1%), post-implantation loss (7.4%) and percent resorptions (7.2%) occurred at a higher incidence than typically seen in the Sprague-Dawley rat (5.9, 5.6 and 5.1%, respectively). The average fetal body weights for both the female and male rats were lower than those typically seen in the Sprague-Dawley rat. External, visceral and skeletal examination of the F1 fetuses revealed numerous malformations and variations which also occurred at higher incidences than those reported for the Sprague-Dawley rat. Routine macroscopic and histomorphologic examination showed there were no changes that would be interpreted to have impaired mating performance, fertility or gestation. Thus, this study provides information on the reproductive effects and the background incidence of embryo-fetal development defects that could be used for comparison to those identified when using this outbred rat for developmental and reproductive toxicity studies, as well as for comparison to the more commonly used rat stock, the Sprague-Dawley rat. For the parameters evaluated, the Wistar Hannover rat had greater variability and an increased incidence of spontaneous malformations as compared to the Crl : CD® (SD)BR Sprague-Dawley rat. These findings should be considered if this stock of rat is selected in the conduct of developmental and reproductive toxicity studies.

Prenatal developmental toxicity studies of 1,1,1‐trichloro‐2,2‐bis(4‐methoxyphenyl)ethane (methoxychlor) in rats and rabbits

ABSTRACT The studies were conducted in rats and rabbits to elucidate the potential developmental toxicity of methoxychlor in general accordance with the improved Japanese MAFF guidelines (12‐Nousan‐No. 8147, 2–1–18, 2000). Methoxychlor dissolved in corn oil was administered orally to pregnant Jcl:SD rats on gestational days (GD) 6–19 at a dose of 0,1,50, or 150 mg/kg/day and to pregnant Kbl:JW rabbits on GD 6–27 at a dose of 0,1, 15, or 45 mg/kg/day. Maternal animals were killed on the day after the last day of administration for morphological examination of their fetuses with special attention to the reproductive organs.Adverse effects on maternal animals were found at 2 higher doses in both species; i.e. vaginal red discharge (rat only), abortion or premature delivery (rabbit only), and significantly decreased body weight gains and food consumption. While the numbers of corpora lutea and implants were comparable between the control and treated groups in both species, slight increase in the percent resorptions and fetal deaths in rats and a reduction of fetal weights in both species were observed at the highest dose. Anogenital distance and testicular histology were not affected in rats. Although fetal examination revealed significant increase in the incidences of 27 presacral vertebrae at 2 higher doses in rabbits, there was no treatment‐related increase in the incidence of malformations in rats and rabbits.Based on these results, it is concluded that methoxychlor causes no malformations, including male reproductive organ abnormalities, in either rats or rabbits, although it results in decreased fetal weights and an increased incidence of fetal resorptions or skeletal variations at maternally toxic doses.