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The nucleotide sequences of the 16S rRNA gene of mitochondrial DNA for all the ape species were determined. On comparing this data with published sequences, all the ape species showed higher diversity than human.

Prostacyclin is an in vivo bioactive substance that regulates renal blood flow (RBF). Information regarding how epoprostenol, a prostacyclin preparation, affects RBF in dogs is lacking. We investigated the effects of short-term epoprostenol administration on RBF in six healthy dogs under anesthesia by administering it intravenously at human doses-2, 5, and 10 ng/kg/min for 20 min. RBF was evaluated before and during epoprostenol administration using pulsed Doppler ultrasonography, and renal perfusion was evaluated using contrast-enhanced ultrasonography. Effects on renal and systemic circulation were evaluated by measuring systolic arterial, mean arterial, diastolic arterial, pulmonary arterial, mean right atrial, and pulmonary capillary wedge pressures; heart rate; and cardiac output. Kruskal-Wallis and Bonferroni multiple comparison tests and Spearman's rank correlation coefficient were used for statistical analyses. As epoprostenol dosage increased, the peak systolic and end diastolic velocity of the renal artery, maximum and minimum venous flow velocities of the interlobular and renal veins, and heart rate all tended to increase, although not significantly. Our results indicate that human-dose epoprostenol administration in dogs does not cause significant changes in renal or systemic circulation. However, the human doses used may have been too low to produce a clinical effect in dogs.

Background: Alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV) is a rare and fatal disorder that occurs in the developing fetal lungs; at birth, infants exhibit an oxygenation disorder accompanied by severe pulmonary hypertension (PH) and have a very short life span. ACDMPV is definitively diagnosed by pathological findings, and infants born with unexplained severe PH may not be properly diagnosed without a biopsy or autopsy., Methods: Japanese infants with unexplained severe PH were enrolled in this study. Genetic analyses were performed on DNA extracted from peripheral blood leukocytes. Sanger sequencing or next-generation sequencing was performed by coding exons and introns for FOXF1 in all samples. For individuals without pathogenic exonic variants, multiplex ligation-dependent probe amplification was performed to identify copy number variations (CNVs) in exons, introns, and in the upstream region of FOXF1., Results: This study included 30 infants who were diagnosed over the course of nine years. Four individuals had the pathogenic variations on the exon 1 of FOXF1, including two frameshift and two missense variations. Pathogenic CNVs were found in another five individuals., Conclusion: In the pathologically proven ACDMPV patients, the ratios of cases with exonic variations, CNVs, and no genetic findings were reported as 45%, 45% and 10%, respectively. We estimate that about 30% (10 (9 + 1) out of 30) of individuals with unexplained severe PH had ACDMPV., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Pulmonary alveolar proteinosis (PAP) is characterized by accumulation of a surfactant-like substance in alveolar spaces and hypoxemic respiratory failure. Genetic PAP (GPAP) is caused by mutations in genes encoding surfactant proteins or genes encoding a surfactant phospholipid transporter in alveolar type II epithelial cells. GPAP is also caused by mutations in genes whose products are implicated in surfactant catabolism in alveolar macrophages (AMs). We performed whole-exome sequence analysis in a family affected by infantile-onset PAP with hypogammaglobulinemia without causative mutations in genes associated with PAP: SFTPB, SFTPC, ABCA3, CSF2RA, CSF2RB, and GATA2. We identified a heterozygous missense variation in OAS1, encoding 2,'5'-oligoadenylate synthetase 1 (OAS1) in three affected siblings, but not in unaffected family members. Deep sequence analysis with next-generation sequencing indicated 3.81% mosaicism of this variant in DNA from their mother's peripheral blood leukocytes, suggesting that PAP observed in this family could be inherited as an autosomal-dominant trait from the mother. We identified two additional de novo heterozygous missense variations of OAS1 in two unrelated simplex individuals also manifesting infantile-onset PAP with hypogammaglobulinemia. PAP in the two simplex individuals resolved after hematopoietic stem cell transplantation, indicating that OAS1 dysfunction is associated with impaired surfactant catabolism due to the defects in AMs., (Copyright © 2018 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)

BackgroundGenetic variants responsible for childhood interstitial lung disease (chILD) have not been studied extensively in Japanese patients.MethodsThe study population consisted of 62 Japanese chILD patients. Twenty-one and four patients had pulmonary hypertension resistant to treatment (PH) and hypothyroidism, respectively. Analyses of genetic variants were performed in all 62 patients for SFTPC and ABCA3, in all 21 PH patients for FOXF1, and in a limited number of patients for NKX2.1.ResultsCausative genetic variants for chILD were identified in 11 (18%) patients: SFTPC variants in six, NKX2.1 variants in three, and FOXF1 variants in two patients. No patients had ABCA3 variants. All three and two patients with NKX2.1 variants had hypothyroidism and developmental delay, respectively. We found six novel variants in this study.ConclusionMutations in SFTPC, NKX2.1, and FOXF1 were identified among Japanese infants and children with chILD, whereas ABCA3 mutations were rare.

Aim: This retrospective study was performed to determine the frequency of malformed infants born at a tertiary center in Hokkaido, Japan. The accuracy of prenatal diagnosis rates was also investigated., Methods: An observational study was performed using data of 1509 and 1743 newborn infants at a single center during two study periods, 2005-2009 (first) and 2010-2014 (second), respectively. Cases including minor anomalies (accessory auricle, nevus and fistula auris congenita) were not included., Results: In total, 274 and 569 malformations were identified in 191 and 337 newborn infants in the first and second study periods, respectively. The number of malformed infants increased significantly over time (13% [191/1509] vs 19% [337/1743], respectively; P < 0.001), mainly as a result of an increase in cases of congenital heart disease (CHD), from 59 to 141 (31% [59/191] vs 42% [141/337] of all malformed infants in the first and second periods, respectively). The overall accurate prenatal diagnosis rate improved over time from 47% (128/274) to 58% (329/569) because of significant improvements in accurate prenatal diagnosis of CHD subtypes (23% [16/70] vs 65% [151/232] in the first and second periods, respectively, P < 0.0001)., Conclusions: The frequency of malformed newborns was higher in the tertiary center than in the general population. The increased number of cases with prenatal suspicion and diagnosis of CHD contributed to the increased frequency of malformed infants during the study period., (© 2016 Japan Society of Obstetrics and Gynecology.)

Previous studies have demonstrated that a light-dark cycle has promoted better sleep development and weight gain in preterm infants than constant light or constant darkness. However, it was unknown whether brief light exposure at night for medical treatment and nursing care would compromise the benefits brought about by such a light-dark cycle. To examine such possibility, we developed a special red LED light with a wavelength of >675 nm which preterm infants cannot perceive. Preterm infants born at <36 weeks' gestational age were randomly assigned for periodic exposure to either white or red LED light at night in a light-dark cycle after transfer from the Neonatal Intensive Care Unit to the Growing Care Unit, used for supporting infants as they mature. Activity, nighttime crying and body weight were continuously monitored from enrolment until discharge. No significant difference in rest-activity patterns, nighttime crying, or weight gain was observed between control and experimental groups. The data indicate that nursing care conducted at 3 to 4-hour intervals exposing infants to light for <15 minutes does not prevent the infants from developing circadian rest-activity patterns, or proper body growth as long as the infants are exposed to regular light-dark cycles.

Background: Among neonates with Down syndrome (DS) and transient leukemia (TL), hyperleukocytosis (white blood cell [WBC] count >100 × 10(9) /L) is associated with increased blood viscosity, respiratory failure due to pulmonary hypertension, multiorgan failure, and increased risk of early death. There have been no previous studies focusing on the effects of exchange transfusion (ExT) on WBC count, respiratory status, and other parameters in TL patients with hyperleukocytosis., Methods: An observational retrospective study was carried out at a single center of all five DS neonates with TL, GATA1 mutations, and hyperleukocytosis, born at a median gestational age of 34 weeks (range, 30-38 weeks) with birthweight 2556 g (range, 1756-3268 g) during a 24 month study period between September 2011 and August 2013. All five neonates underwent ExT at a median age of 2 days (range, 0-5 days) before initiation of other cytoreductive therapy with cytarabine, which was carried out in two patients., Results: All patients required respiratory support before ExT. After ExT, respiration status improved in all five patients: WBC count (mean) decreased by 85% from 143 × 10(9) /L to 21 × 10(9) /L. None developed tumor lysis syndrome. Three survived and two died: one hydrops fetalis neonate born at gestational week 30 died at age 5 days, and another died eventually from acute gastroenteritis 40 days after leaving hospital at the age of 155 days with complete remission. Two of the three surviving neonates developed acute megakaryocytic leukemia at age 90 days and 222 days., Conclusion: ExT was very effective in improving hyperleukocytosis and may have had favorable effects on respiration., (© 2015 Japan Pediatric Society.)

Background: Genetic variations associated with interstitial lung diseases (ILD) have not been extensively studied in Japanese infants., Methods: Forty-three infants with unexplained lung dysfunction were studied. All 43, 22, and 17 infants underwent analyses of surfactant protein (SP)-C gene (SFTPC) and ATP-binding cassette A3 gene (ABCA3), SP-B gene (SFTPB), and SP-B western blotting, respectively. Two and four underwent assessment of granulocyte macrophage colony-stimulating factor-stimulating phosphorylation of signal transducer and activator of transcription-5 (pSTAT-5) and analyses of FOXF1 gene (FOXF1), respectively., Results: ILD were diagnosed clinically in nine infants: four, three, and two had interstitial pneumonitis, hereditary pulmonary alveolar proteinosis (hPAP), and alveolar capillary dysplasia with misalignment of pulmonary veins (ACD/MPV), respectively. Genetic variations considered responsible were detected in six (67%) of the nine infants with ILD: three with hPAP (SFTPC p.Leu45Arg and p.Gln145fs, and ABCA3 p.Arg1583Trp/p.Val1495CysfsX21), two with interstitial pneumonitis (SFTPC p.Lys63Glu and p.Ser72Asn/p.Gly100Ala), and one with ACD/MPV (FOXF1 p.Leu300ArgfsX79). None showed SFTPB mutations or defects in pSTAT-5. The 17 bronchoalveolar lavage or tracheal aspirates contained enough SP-B protein., Conclusion: The SP-C abnormality was most prevalent, and SP-B deficiency was rare in Japanese infants with hereditary ILD.

In the brain, serotonin production is controlled by tryptophan hydroxylase 2 (TPH2), a genotype. Previous studies found that mutations on the TPH2 locus in humans were associated with depression and studies of mice and studies of rhesus macaques have shown that the TPH2 locus was involved with aggressive behavior. We previously reported a functional single nucleotide polymorphism (SNP) in the form of an amino acid substitution, Q468R, in the chimpanzee TPH2 gene coding region. In the present study we tested whether this SNP was associated with neuroticism in captive and wild-born chimpanzees living in Japan and Guinea, respectively. Even after correcting for multiple tests (Bonferroni p = 0.05/6 = 0.008), Q468R was significantly related to higher neuroticism (β = 0.372, p = 0.005). This study is the first to identify a genotype linked to a personality trait in chimpanzees. In light of the prior studies on humans, mice, and rhesus macaques, these findings suggest that the relationship between neuroticism and TPH2 has deep phylogenetic roots.

Nine male chimpanzees originally reared in solitary cages were set up to form a group. Plasma viral load of the lymphocryptovirus (LCV) of chimpanzee [Epstein-Barr virus chimpanzee (EBVcmp)] was measured by real-time PCR. In the group formation (Form) period, the first-ranking male showed an imminent increase in plasma EBVcmp load compared with 1 week before (pre-Form) and 3 months after (post-Form) group formation. Other upper-ranking males such as the second-, third- and fourth-male also showed the highest level of viral load in the Form period. The kinetics of EBVcmp load in the Form period were statistically different from other periods (against pre-Form, t=-4.878, P<0.001; against post-Form, t=6.434, P<0.001). The effect of the male dominance rank did not differ between the pre-Form and post-Form periods (t=-1.557, P=0.12). Reactivation of LCV (EBV) as an immunological stress marker for humans might also be applied to chimpanzees.

Owing to its high temporal sensitivity, saliva has distinct advantages for measuring steroids, compared with other noninvasive samples such as urine and feces. Here, we report the validity of assaying salivary cortisol (C) and testosterone (T) using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in captive male chimpanzees, Pan troglodytes. For both the C and T concentrations, we found positive relationships between saliva and plasma. The concentrations of C and T in saliva showed clear patterns of diurnal fluctuation, whereas those in urine and feces did not. These results suggest that the salivary steroid concentrations can be regarded as good indicators of circulating steroid levels. We also developed and validated an efficient method for collecting saliva samples from cotton rope. Although rope includes inherent steroid-like compounds and may affect the accuracy of steroid measurements, our rope-washing procedures effectively removed intrinsic steroidal materials. There was a significant association between the C and T concentrations measured from saliva collected from rope licked by the chimpanzees and those measured from saliva collected directly from the mouth. Salivary T values estimated by LC/MS-MS were similar to those measured by radioimmunoassay. The results indicate the usefulness of saliva as a noninvasive steroid measure and that steroids in the saliva of chimpanzees can be accurately measured by LC-MS/MS.

We surveyed repeat sequence in the 5' flanking region of primate arginine vasopressin receptor 1a gene (AVPR1a) which is important for social behavior in mammals. The amplified region was polymorphic in length in all species investigated. In addition, length variants were examined for promoter activity by in vitro expression of the luciferase gene. However, no significant difference was observed. This region could be a marker to further survey functional difference between and within species.

The main focus of this study is to determine the optimal administration period concerning toxic effects on ovarian morphological changes in a repeated-dose toxicity study. To assess morphological and functional changes induced in the ovary by bromocriptine, the compound was administered to female rats at dose levels of 0, 0.08, 0.4 and 2 mg/kg for the 2- or 4-week repeated-dose toxicity study, and for the female fertility study from 2 weeks prior to mating to day 7 of gestation. In the 2-week repeated-dose toxicity study, increase of ovarian weights was observed at 2 mg/kg. In the 4-week repeated-dose toxicity study, ovarian weights were increased at 0.4 and 2 mg/kg. The number of corpora luteum was increased in the 0.4 and 2 mg/kg groups of the 2- and 4-week repeated-dose toxicity studies by histopathological examination of the ovaries. Bromocriptine did not affect estrous cyclicity in 2- and 4-week repeated dosing. In the female fertility study, although animals in any groups mated successfully, no females in 0.4 and 2 mg/kg groups were pregnant. There were no adverse effects on reproductive performance in the 0.08 mg/kg group. Based on these findings, the histopathological changes in the ovary are considered important parameters for evaluation of drugs including ovarian damage. We conclude that a 2-week administration period is sufficient to detect ovarian toxicity of bromocriptine in a repeated-dose toxicity study.

Monoamine oxidases (MAOs) are involved in the degradation of various biogenic amines such as dopamine and serotonin. Two isozymic genes, MAOA and MAOB, have been identified, both of which are located on the X chromosome. In humans, intron 2 of both MAOs contains dinucleotide repeats, MAin2 and MBin2, that have been suggested as candidate genes for mood disorders and Parkinson's disease. In this study, we surveyed the corresponding regions in nonhuman primates. The MAin2 and MBin2 regions were amplified in a total of 37 species from all primate lineages, including samples from 6 simian and 7 prosimian species. MAin2 and MBin2 were successfully amplified for all the simian species but failed to amplify in the prosimians. A series of alleles of MAin2 and MBin2 ranging from 222 bp to 254 bp and from 224 bp to 284 bp, respectively, were identified. Chimpanzees, gorillas, orangutans, and gibbons were polymorphic for both MAin2 and MBin2. Common marmosets and tamarins were polymorphic for MAin2 only. In the genus Macaca, MBin2 was polymorphic in 15 of the 18 species examined and 2-11 alleles were observed. Conversely, MAin2 was not diverse, exhibiting only 1-2 alleles. Since the MAin2 and MBin2 regions are associated with mental disorders, they might serve as useful markers for surveying the association with behavioral traits in the abovementioned primate species.

A variable number of tandem repeat (VNTR) polymorphism based on a 16 or 17-bp unit has been reported in the third intron of the human serotonin transporter gene (5-HTT). VNTRs have been shown to affect the transcriptional activity of genes, and VNTR polymorphisms possibly influence human personality and several psychoneurological disorders. To estimate the changes that occurred in the VNTRs during primate evolution, we amplified and sequenced the regions that corresponded to the human VNTRs in various primate species, including apes, Old World monkeys, and New World monkeys. The VNTR sequences were polymorphic in all the ape species examined, and alleles with repeat numbers of 18, 19, 23, and 24 in chimpanzees, 33, 35, 36, 38, and 40 in gorillas, 4 and 6 in orangutans, and 11, 13, 14, and 15 in gibbons were found. On the other hand, only a 5-repeat allele was detected in Old World monkeys such as the Japanese macaque and patas monkey. In this study we demonstrated for the first time that a repeat structure was not present in the corresponding regions in the New World monkeys examined, and only one unit sequence was found in them. These results suggested that the duplication of a unit in the VNTR region occurred in the Cercopithecidae species following the divergence of the Old World and New World monkeys, and various long repeated alleles were generated in humans and apes, except orangutans.

The genetic basis of the phenotypic difference between human and chimpanzee is one of the most actively pursued issues in current genomics. Although the genomic divergence between the two species has been described, the transcriptomic divergence has not been well documented. Thus, we newly sequenced and analyzed chimpanzee full-length cDNAs (FLcDNAs) representing 87 protein-coding genes. The number of nucleotide substitutions and sites of insertions/deletions (indels) was counted as a measure of sequence divergence between the chimpanzee FLcDNAs and the human genome onto which the FLcDNAs were mapped. Difference in transcription start/termination sites (TSSs/TTSs) and alternative splicing (AS) exons was also counted as a measure of structural divergence between the chimpanzee FLcDNAs and their orthologous human transcripts (NCBI RefSeq). As a result, we found that transposons (Alu) and repetitive segments caused large indels, which strikingly increased the average amount of sequence divergence up to more than 2% in the 3'-UTRs. Moreover, 20 out of the 87 transcripts contained more than 10% structural divergence in length. In particular, two-thirds of the structural divergence was found in the 3'-UTRs, and variable transcription start sites were conspicuous in the 5'-UTRs. As both transcriptional and translational efficiency were supposed to be related to 5'- and 3'-UTR sequences, these results lead to the idea that the difference in gene regulation can be a major cause of the difference in phenotype between human and chimpanzee.

Polymorphic microsatellite repeats in the promoter region of estrogen receptor alpha gene (ESRalpha and the intron 6 region of estrogen receptor beta gene (ESRbeta) have been reported in human populations. To examine the evolutional state of both repeats, we surveyed the corresponding regions in DNA sequences from the following great apes and gibbons: 56 chimpanzees, 3 bonobos, 16 gorillas, 20 orangutans and 60 gibbons (four species: 17 of Hylobates agilis, 11 of H. lar, 15 of H. muelleri, and 17 of H. syndactylus). In the corresponding region of the TA repeat of human ESRalpha, chimpanzees and bonobos had two motifs in the repeat tract, (TA)(7-9) and (CA)(4-6). Gorillas had the (TA)(9-10) repeat tracts and orangutans had monomorphic (TA)(7) repeats. Although all great apes maintained the TA expansion, all gibbon sequences contained (TA)(2), implying that the CA dinucleotide expansion arose in the ancestor of chimpanzees and bonobos. The nucleotide sequences of ESRbeta showed a very complex repeat pattern in apes. The human sequences had a non-variable preceding sequence at (CA)(n), (GA)(2)(TA)(8)(CA)(4)(TA). In apes that region included {(TA)(n)(CA)(n)}(n). Gibbon sequences included (TATG)(n) and (TATC)(n) and no regular construction was observed. A deletion event in the reverse primer site seems to have occurred in the orangutan lineage. In addition, a great diversity of allele length was detected in each gibbon species.

We examined the adolescent development of the sexual skin of chimpanzees (Pan troglodytes) by daily observation, evaluation of swelling, and weekly photogrametry. Although the size of swelling differed with the individual, the development of sexual swelling followed four stages: (1) initial stage, the labial region began to show a slight swelling and recovery; (2) labial stage, swelling at the labial region became maximal; (3) anal stage, another swelling center appeared in the anal region and enlarged; and (4) full maturity stage, the labial and anal regions merged into a full swelling. Menarche occurred after the beginning of the anal stage, and the regular cycle was then established. All of the swelling stages and the peak swelling size are regarded as good indicators of reproductive maturation in chimpanzees.

Tryptophan hydroxylase 2 (TPH2) is a rate-limiting enzyme of neuronal serotonin biosynthesis. Recently, two single nucleotide polymorphisms (SNPs) at the exon 11 coding region that resulted in amino acid substitutions in the C-terminal domain have been reported to affect enzyme activity in humans and mice. We determined 175 base-pair sequences of the exon 11 region in nine primate species from all recognized lineages. All nucleotide sequence substitutions were synonymous, with the exception of one adenine (A) to guanine (G) substitution at the 1404th position in the open reading frame (ORF). This substitution leads to a glutamine (Q) to arginine (R) amino acid substitution at the 468th position within chimpanzee sequences. The frequency of the G allele was 0.24 among 66 chimpanzees. Therefore, it is a novel SNP observed in chimpanzees, and we have named these two alleles as ch468Q and ch468R, respectively. When expressed in HeLa cells, ch468R caused an approximate 20% increase in enzyme function during L-5-hydroxytryptophan (5HTP) production (P<0.001). We also surveyed the interspecies difference in enzyme activity among human, chimpanzee, and rat. Although the rat showed an identical amino acid sequence at the C-terminal region as those of human and ch468Q, the rat enzyme was more active than those of human or chimpanzee (P<0.001), indicating the importance of substitutions in other regions. Our findings on the chimpanzee SNP will be a useful genetic marker in understanding the individual difference in the serotonin-related behavior.

A variable number of tandem repeats (VNTR) polymorphism based on a 30-bp unit have been reported in the promoter region of the human monoamine oxidase A gene (MAOA). Human VNTRs have been shown to affect transcriptional activity, and some reports suggest that VNTR polymorphisms are associated with psychoneurological disorders. VNTR polymorphism has also been reported in the ape MAOA promoter but the transcriptional activities of the alleles remain to be determined. In the present study, we sequenced the 1.3-kb promoter region of ape MAOA and compared the transcriptional activities of ape MAOA promoter sequences with those of humans. All apes examined were polymorphic in the region corresponding to the human VNTR and two, four, three, and two alleles were found in chimpanzees, gorillas, orangutans, and gibbons, respectively. VNTR repeat structures in gorillas, orangutans, and gibbons were considerably different from those in humans and chimpanzees. In a human neuroblastoma cell line, most of the ape sequences that had a short repeat length (12bp or 18bp) exhibited higher promoter activity than a human 3-repeat sequence with a 30-bp repeat length. However, an intra-species difference dependent on the repeat number was not observed among the ape alleles examined.

We examined the toxicokinetics of bisphenol A (BPA) in F344 rats, cynomolgus monkeys and chimpanzees. Serum BPA levels were quantified using the LC-MS/MS method. After oral administration at 10 mg/kg, the maximum concentration in the serum (C(max)) and the area under the serum concentration curve (AUC) of BPA in cynomolgus monkeys and chimpanzees were greater than in rats. After oral administration at 100 mg/kg, AUC during the first 4h (AUC(0-->4h)) in cynomolgus monkeys was greater than in rats. In rats, the serum BPA levels were increased again 6h or later after oral administration at each dose, which suggested the enterohepatic circulation of BPA in rats. After subcutaneous administration at 10 mg/kg, the AUCs were ranked in the following order: cynomolgus monkeys>chimpanzees>rats, and C(max) in cynomolgus monkeys was greater than in rats and chimpanzees. After subcutaneous administration at 100 mg/kg to cynomolgus monkeys and rats, both the C(max) and AUCs in cynomolgus monkeys were greater than in rats. In all species, the oral administration of BPA resulted in much lower C(max) and AUCs than subcutaneous administration at the corresponding doses, indicating the low bioavailability of oral administration. This result suggests that BPA undergoes an extensive first-pass metabolism in these animal species. AUCs of subcutaneous administration and the AUC (0-->4h) of oral administration in the two primates were greater than that in rats. Because the systemic clearance for BPA is assumed to be dependent on the hepatic blood flow-rate, the high AUCs in primates are considered to be due to the lower systemic clearance by a lower hepatic blood flow-rate in primates than in rats. In addition, the toxicokinetics of the metabolites of BPA were examined. After the oral administration of 10 mg/kg BPA, both C(max) and AUCs of BPA metabolites were ranked in the following order: cynomolgus monkeys>chimpanzees>rats, and the terminal elimination half-life (T(1/2)) in rats was greater than that in cynomolgus monkeys and chimpanzees, suggesting the enterohepatic circulation of BPA in rats. From these results, the systemic clearance of BPA in primates is considered to be close to that in humans due to the similarity of the hepatic blood flow-rate. Furthermore, the major elimination route of BPA metabolites in primates is assumed to be renal excretion, as in humans, because the enterohepatic circulation that was observed in rats was not observed. In conclusion, primates are thought to be served as a valuable surrogate model for the toxicokinetics of BPA in humans.

The major histocompatibility complex (MHC) class-II genes are described as the genes that encode the antigen-presenting molecule. In particular, functional alleles of MHC class-II genes in nonhuman primates have been analyzed as part of the study of infectious and immune-mediated diseases in animal models and as part of efforts to understand the molecular evolution of human leukocyte antigen. The polymorphisms and sequence analysis of MHC class-II genes in a large number of subjects is necessary for the group management of nonhuman primates. In the present study, we attempted to analyze the DQB1 polymorphism in the common chimpanzee (Pan troglodytes) by exon 2 sequencing. For exact typing of Patr-DQB1 alleles, we carried out polymerase chain reaction-restriction fragment length polymorphism and sequence analysis. In the genomic DNA of 25 chimpanzees, 6 Patr-DQB1 alleles, including 2 new alleles, were detected. Identification and analysis of Patr-DQB1 alleles using this method may contribute to our understanding of Patr-DQB1 allelic diversity in individual chimpanzees and should be useful in facilitating colony management of chimpanzee groups in Japan.

Two polymorphic trinucleotide repeats of human androgen receptor gene (hAR), CAG and GGN which encode glutamine and glycine, have been shown to be associated with human diseases. The number of repeats ranges from 8 to 35 for the CAG and from 10 to 30 for the GGN in human populations. Longer CAG repeats are associated with reduced hAR transcriptional activity, spinal bulbar muscular atrophy and lower cognitive function in older men, whereas shorter CAG repeats are associated with increased risk of prostate cancer and infertility in men. The functional roles of the CAG and GGN repeats have not been clarified. In order to compare the sequence of the CAG and GGN regions in apes, we analyzed 57 chimpanzees, 18 gorillas, 20 orangutans, 16 agile gibbons, and 17 siamangs by PCR and electrophoresis. Two bonobos and one long-tailed macaque were also sequenced and the sequences of all species were aligned, respectively, with one human registered sequence. Seventeen different alleles (4, 7, 8, 9, 12, 14, 15, and 17-26 repeats) and 11 alleles (11-14 and 16-22 repeats) were detected at the CAG and the GGN loci, respectively. Although the repeat tract was conserved among apes, chimpanzees had alleles with a wide range of repeat lengths: (CAG)(14-26) and (GGN)(14-22). Gorillas were less polymorphic with the (CAG)(8) and (GGN)(19) alleles being most common, and orangutans exhibited monomorphic (CAG)(11) and (GGN)(22) alleles. On the other hand, agile gibbons and siamangs had the shortest (CAG)(4) allele, but showed variable length of GGN repeats (11-13 in agile gibbons and 16-21 in siamangs). In chimpanzees, frequent haplotypes consisting of short CAG repeats and long GGN repeats or vice versa was observed as in humans.

We attempted ovarian stimulation using gonadotropins in 14 chimpanzees. Subjects were given a single administration of leuprorelin acetate, followed by repeated administration of human menopausal gonadotropin (hMG) for 16-21 days. During the dosing period, the ovarian follicle diameter and count were measured by transvaginal ultrasonography. The hormone administration induced the development of multiple follicles, and multiple oocytes were subsequently retrieved. However, the follicle count was decreased, suggesting atresia, in some subjects. Statistically, the final follicle diameter was dependent on the dosing duration and the hMG dose in the late stage, while the maximum follicle count during hMG administration was dependent on age and the hMG dose in the early stage. Five subjects showed mild ovarian hyperstimulation syndrome (OHSS)-like symptoms with a high serum estradiol (E2) concentration. These results suggest that leuprorelin acetate plus hMG administration successfully stimulates the development of multiple ovarian follicles for oocyte retrieval and that the serum E2 concentration is predictive of OHSS-like symptoms in chimpanzees.

Chimpanzees and macaques were compared in their growth of head, face and body, based on a large-scale somatometrical database. Their growth stages, i to v, were determined by inflection points in velocity curves. Sex differences in their growth are shown both by elongated stages in juvenile and adolescent and by the greater velocity in males in the stages. Chimpanzees need longer to get their full growth, especially in the later infantile and juvenile stages. The growth patterns are classified into three types of "sigmoid", "parabolic", and "fast & slow" in distance curves, and in velocity curves, they correspond to "convex", acceleration in mid-growth stage; "linear", linear deceleration with age; and "concave", rapid deceleration in earlier stages and slow velocity in later stages. Great differences between chimpanzees and macaques were found in their growth patterns of upper facial height and facial height, which are "linear" or intermediate of "linear" and "concave" in macaques and "concave" in chimpanzees. These differences in their growth patterns explain the characteristic development of craniofacial proportions.

Several human neurodegenerative disorders are caused by the expansion of polymorphic trinucleotide repeat regions. Many of these loci are functional short tandem repeats (STRs) located in brain-expressed genes, and their study is thus relevant from both a medical and an evolutionary point of view. The aims of our study are to infer the comparative pattern of variation and evolution of this set of loci in order to show species-specific features in this group of STRs and on their potential for expansion (therefore, an insight into evolutionary medicine) and to unravel whether any human-specific feature may be identified in brain-expressed genes involved in human disease. We analyzed the variability of the normal range of seven expanding STR CAG/CTG loci (SCA1, SCA2, SCA3-MJD, SCA6, SCA8, SCA12, and DRPLA) and two nonexpanding polymorphic CAG loci (KCNN3 and NCOA3) in humans, chimpanzees, gorillas, and orangutans. The study showed a general conservation of the repetitive tract and of the polymorphism in the four species and high heterogeneity among loci distributions. Humans present slightly larger alleles than the rest of species but a more relevant difference appears in variability levels: Humans are the species with the largest variance, although only for the expanding loci, suggesting a relationship between variability levels and expansion potential. The sequence analysis shows high levels of sequence conservation among species, a lack of correspondence between interruption patterns and variability levels, and signs of conservative selective pressure for some of the STR loci. Only two loci (SCA1 and SCA8) show a human specific distribution, with larger alleles than the rest of species. This could account, at the same time, for a human-specific trait and a predisposition to disease through expansion.

Prior to fertilization by intracytoplasmic sperm injection (ICSI), cytoplasmic organization was evaluated in metaphase II chimpanzee oocytes obtained from stimulated ovaries. The findings demonstrate a high frequency of anomalies that are remarkably similar to the types of cytoplasmic dysmorphisms reported for human oocytes used in IVF. Similar to the human, the occurrence of these anomalies was oocyte- and animal-specific and associated with reduced competence as indicated by embryo development in vitro to the blastocyst stage.

We compared the toxicokinetics of bisphenol A (BPA) among three animal species: rats, cynomolgus monkeys and chimpanzees. Rats and monkeys were administered BPA orally or subcutaneously at 10 or 100 mg/kg body weight, while chimpanzees were administered only 10 mg/kg of BPA. BPA in serum was measured by ELISA. In oral administration of BPA at 10 mg/kg, both C(max) and AUC were rats < chimpanzee < monkeys. In oral administration of BPA at 100 mg/kg, both C(max) and AUC were rats < monkeys. Subcutaneous BPA administrations also revealed similar results, although the values of toxicokinetic parameters in subcutaneous administration were higher than those in oral administration. These results suggest that orally or subcutaneously administered BPA in primates is more easily absorbed than that in rats. We conclude that there are considerable differences in distribution, metabolism, and excretion of BPA between rodents and primates.

Several lines of evidence suggest that patterns of genetic variability in the human angiotensinogen gene (AGT) contribute to phenotypic variability in human hypertension. The A(-6) promoter variant of AGT is associated with higher plasma angiotensinogen levels and increased risk of essential hypertension. The geographic distribution of the A(-6) variant leads to the intriguing hypothesis that the G(-6) promoter variant has been selectively advantageous outside Africa. To test these hypotheses, we investigated the roles of population history and natural selection in shaping patterns of genetic diversity in AGT, by sequencing the entire AGT gene (14400 bp) in 736 chromosomes from Africa, Asia, and Europe. We found that the A(-6) variant is present at higher frequency in African populations than in non-African populations. Neutrality tests found no evidence of a departure from selective neutrality, when whole AGT sequences were compared. However, tests restricted to sites in the vicinity of the A(-6)G polymorphism found evidence of a selective sweep. Sliding-window analyses showed that evidence of the sweep is restricted to sites in tight linkage disequilibrium (LD) with the A(-6)G polymorphism. Further, haplotypes carrying the G(-6) variant showed elevated levels of LD, suggesting that they have risen recently to high frequency. Departures from neutral expectation in some but not all regions of AGT indicate that patterns of diversity in the gene cannot be accounted for solely by population history, which would affect all regions equally. Taken together, patterns of genetic diversity in AGT suggest that natural selection has generally favored the G(-6) variant over the A(-6) variant in non-African populations. However, important localized effects may also be present.

The dopamine receptor D4 (DRD4) has received increasing research attention in behavioral science, psychiatry, and psychopharmacology. However, the number of available genetic markers for primates is still insufficient. We identified a novel variation/polymorphism in the second intron of DRD4 in humans based on the survey of 210 Japanese: a 6bp insertion (allele frequency: 0.002) and 8bp deletion (0.024); however, 94 Hungarian Caucasians were found to be monomorphic. Polymorphisms of the homologous region were also found in a survey of 93 specimens from four species of great apes and 51 specimens from seven species of gibbons. The polymorphisms consist of both single nucleotide substitutions and variations in the number of tandem duplications of short GC-rich sequences. Because of usefulness of primates in behavioral science, this polymorphism may be a useful marker for association studies with behavioral traits in both humans and apes.

Ubiquitin is a highly conserved protein, and is encoded by a multigene family among eukaryote species. The polyubiquitin genes, UbB and UbC, comprise tandem multiple ubiquitin coding units without a spacer region or intron. We determined nucleotide sequences for the UbB and UbC of human, chimpanzee, gorilla, and orangutan. The ubiquitin repeat number of UbB was constant (3) in human and great apes, while that of UbC varied: 6 to 11 for human, 10 to 12 for chimpanzee, 8 for gorilla, and 10 for orangutan. The heterogeneity of the repeat number within closely related hominoid species suggests that a lineage-specific unequal crossover and/or gene duplication occurred. A marked homogenization of UbC occurred in gorilla with a low level of synonymous difference (p(s)). The homogenization of UbC also occurred in chimpanzee and less strikingly in human. The first and last ubiquitin coding units of UbC were clustered independently between species, and less affected by homogenization during the hominoid evolution. Therefore, the homogenization of ubiquitin coding units is likely due to an unequal crossing-over inside the ubiquitin units. The lineage-specific homogenization of UbC among closely related species suggests that concerted evolution has a key role in the short-term evolution of UbC.

We constructed full-length enriched cDNA libraries from chimpanzee brain, skin, and liver tissues by the oligo-capping method to establish a database of sequences of chimpanzee genes. Randomly selected clones from the libraries were subjected to one-pass sequencing from their 5'-ends. As a result, we collected 6813 chimpanzee cDNA sequences longer than 400 bp. Homology search against human mRNA sequences (RefSeq mRNAs) revealed that our collection included sequences of 1652 putative chimpanzee genes. In order to calculate the sequence identity between human and chimpanzee homologs, we constructed 5'-end consensus sequences of 226 chimpanzee genes by aligning at least three sequences for individual genes. Sequence identity was estimated by comparing these consensus sequences and the corresponding sequences of their human homologs. The average sequence identity of the 5'-end cDNAs was 99.30%. Those of the 5'-UTRs and CDSs were 98.79% and 99.42%, respectively. The results confirmed that human and chimpanzee genes are highly conserved at the nucleotide level. As for amino acids, the average sequence identity was 99.44%. The average synonymous (K(S)) and nonsynonymous (K(A)) divergences were estimated to be 1.33% and 0.28%, respectively.