Environmental enrichment-related injury in a macaque (Macaca fascicularis): intestinal linear foreign body

A three-year old male cynomolgus macaque (Macaca fascicularis) presented with clinical signs of anorexia and depression that decreased over a 48-hour period. Results of abdominal radiography abdominocentesis, blood biochemical analysis and CBC suggested septic peritonitis. Exploratory laparotomy revealed multiple perforations along the mesenteric border of the small intestine. Necropsy revealed masses of fibrous material in the stomach and cecum. Multiple mucosal ulcerations, as well as linear fibrous material, were found in the small intestine. The ulceration, perforations, and septic peritonitis were attributed to the ingestion of rope that had been attached to the animal's cage as an environmental-enrichment device.     

H5N1禽流感病毒感染小鼠和两种灵长类动物的肺组织病理学比较

目的比较H5N1禽流感病毒感染小鼠、恒河猴及食蟹猴急性期肺组织的病理学变化。方法在麻醉状态下对BALB／c小鼠、恒河猴及食蟹猴进行H5N1病毒滴鼻接种,在感染急性期实施安死术,取肺组织运用H＆E结合免疫组化技术分析肺组织的病理变化。结果BALB／c小鼠感染急性期,肺组织以变质性炎为主,肺泡结构被广泛破坏,以单核细胞为主的炎细胞浸润,局部可见渗出性炎。而在恒河猴感染急性期肺组织病理改变以渗出性炎为主,同时可见变质性炎和增生性炎。在食蟹猴感染急性期肺组织病理改变以渗出性和变质性炎为主,同时亦可见上皮的新生。结论H5N1禽流感病毒感染小鼠与恒河猴、食蟹猴急性期肺组织的病理变化不同,这将为进一步认识禽流感的发病机制及研究针对性的治疗方法提供一些理论依据。     

神经皮肤黑变病中黑色素的来源初探

目的探讨食蟹猴神经皮肤黑变病（NCM）中黑色素的来源。方法收集北京协和医学院比较医学中心病理室食蟹猴神经皮肤黑变病的病理学标本一例,复习其病理切片并辅助免疫组化及电镜技术对黑色素的来源做进一步探讨。结果皮肤病理检查显示真皮内黑色素主要由黑色素细胞产生,黑色素细胞CD31免疫组化染色阴性。脑膜的组织病理学显示脑膜下黑色素主要由黑色素细胞产生,黑色素细胞CD31免疫组化染色阴性。脑实质的组织病理学显示其黑色素由黑色素细胞和血管内皮细胞产生,CD31免疫组化染色显示含有黑色素的血管内皮细胞呈阳性着色。电镜检查结果同样显示血管内皮细胞内可见黑色素小体沉积。结论我们在世界上首次揭示了食蟹猴NCM大脑中血管内皮细胞可产生黑色素,这给内皮细胞增加了一项新的功能。     

Cutaneous melanoma with metastasis in a cynomolgus monkey (Macaca fascicularis)

Background  A 3.3-year-old-male cynomolgus macaque ( Macaca fascicularis ) showed a focally extensive soft, dark, discoid dermal mass, 0.5 cm in diameter, on the dorsal surface of the right hind foot, over the fourth and fifth metatarsal bones.
Methods and results  Microscopic examination revealed a cutaneous melanoma with local lymphatic invasion, characterized by neoplastic melanocytes within the subcapsular sinus of popliteal and inguinal lymph nodes. The diagnosis was confirmed by immunohistochemistry and transmission electron microscopy.
Conclusions  To our knowledge, this is the first documented case of melanoma in a cynomolgus monkey.     

Whole-genome sequencing and analysis of the Malaysian cynomolgus macaque (Macaca fascicularis) genome

ABSTRACT: BACKGROUND: The genetic background of the cynomolgus macaque (Macaca fascicularis) is made complex by the high genetic diversity, population structure, and gene introgression from the closely related rhesus macaque (Macaca mulatta). Herein we report the whole-genome sequence of a Malaysian cynomolgus macaque male with more than 40-fold coverage, which was determined using a resequencing method based on the Indian rhesus macaque genome. RESULTS: We identified approximately 9.7 million single nucleotide variants (SNVs) between the Malaysian cynomolgus and the Indian rhesus macaque genomes. Compared with humans, a smaller nonsynonymous/synonymous SNV ratio in the cynomolgus macaque suggests more effective removal of slightly deleterious mutations. Comparison of two cynomolgus (Malaysian and Vietnamese) and two rhesus (Indian and Chinese) macaque genomes, including previously published macaque genomes, suggests that Indochinese cynomolgus macaques have been more affected by gene introgression from rhesus macaques. We further identified 60 nonsynonymous SNVs that completely differentiated the cynomolgus and rhesus macaque genomes, and that could be important candidate variants for determining species-specific responses to drugs and pathogens. The demographic inference using the genome sequence data revealed that Malaysian cynomolgus macaques have experienced at least three population bottlenecks. CONCLUSIONS: This list of whole-genome SNVs will be useful for many future applications, such as an array-based genotyping system for macaque individuals. High-quality whole-genome sequencing of the cynomolgus macaque genome may aid studies on finding genetic differences that are responsible for phenotypic diversity in macaques and may help control genetic backgrounds among individuals.     

Fibrous Dysplasia of Bone Associated with Primary Hyperparathyroidism

ObjectiveFibrous dysplasia of bone and primary hyperparathyroidism (PHPT) may occur in patients with McCune-Albright Syndrome. A small number of cases with both diagnoses that are not associated with the above-mentioned genetic disorder have been published in the literature. It is uncertain if these disorders are linked in some way. In the present study, we aimed to further explore a potential relationship between PHPT and fibrous dysplasia of bone.MethodsWe conducted a retrospective review of all cases seen at Mayo Clinic, Rochester, Minnesota, between 1976 and 2011 that were diagnosed with both PHPT and fibrous dysplasia of bone.ResultsWe identified 10 patients who were diagnosed with both PHPT and fibrous dysplasia of bone. Fibrous dysplasia was polyostotic in 7 (70%) cases. It affected the lower extremities in 6 (60%) patients, the skull or facial bones in 4 (40%), and was localized to one rib in 1 patient (10%). In 4 patients, fibrous dysplasia was diagnosed first, between 9 to 50 years before being diagnosed with PHPT. Two cases of fibrous dysplasia were recognized between 2 and 5 years after the diagnosis of PHPT. The remaining 4 patients were diagnosed with both conditions at approximately the same time.ConclusionsIt remains unclear if the association between fibrous dysplasia of bone and PHPT is more than coincidental, although the possibility of a rare familial genetic syndrome is not completely excluded.     

Genome sequencing and comparison of two nonhuman primate animal models, the cynomolgus and Chinese rhesus macaques

The nonhuman primates most commonly used in medical research are from the genus Macaca. To better understand the genetic differences between these animal models, we present high-quality draft genome sequences from two macaque species, the cynomolgus/crab-eating macaque and the Chinese rhesus macaque. Comparison with the previously sequenced Indian rhesus macaque reveals that all three macaques maintain abundant genetic heterogeneity, including millions of single-nucleotide substitutions and many insertions, deletions and gross chromosomal rearrangements. By assessing genetic regions with reduced variability, we identify genes in each macaque species that may have experienced positive selection. Genetic divergence patterns suggest that the cynomolgus macaque genome has been shaped by introgression after hybridization with the Chinese rhesus macaque. Macaque genes display a high degree of sequence similarity with human disease gene orthologs and drug targets. However, we identify several putatively dysfunctional genetic differences between the three macaque species, which may explain functional differences between them previously observed in clinical studies.     

Frequency of spontaneous congenital defects in rhesus and cynomolgus macaques

Abstract: This paper summarizes the spontaneous incidence of congenital defects in the rhesus and cynomolgus macaque colonies (Macaca mulatta and M. fascicularis) at the California Regional Primate Research Center. The computerized database used in this analysis included fetuses, term infants, juveniles, and adults that underwent a necropsy procedure over a 14-year period (1983–1996). The calculated malformation rates were 0.9% (40/4,390) and 0.3% (3/965) for the rhesus and cynomolgus monkey, respectively. Most of the observed malformations in both species affected the musculoskeletal and the cardiovascular systems, while a smaller number of defects were observed in the gastrointestinal, urogenital, endocrine, and central nervous systems. Inbreeding did not contribute to the spontaneous malformation incidence and there was no predilection for sex (male vs. female) or housing (indoors vs. outdoors) among the malformed cases. This spontaneous malformation database in our macaque colony aids in the interpretation of defects that occur in an experimental study as well as in the ongoing assessment of a healthy nonhuman primate breeding colony.     

Molecular Evidence for Distinct Genotypes of Monkey B Virus (Herpesvirus Simiae) Which Are Related to the Macaque Host Species

Although monkey B virus (herpesvirus simiae; BV) is common in all macaque species, fatal human infections appear to be associated with exposure to rhesus macaques (Macaca mulatta), suggesting that BV isolates from rhesus monkeys may be more lethal to nonmacaques than are BV strains indigenous to other macaque species. To determine if significant differences that would support this supposition exist among BV isolates, we compared multiple BV strains isolated from rhesus, cynomolgus, pigtail, and Japanese macaques. Antigenic analyses indicated that while the isolates were very closely related to one another, there are some antigenic determinants that are specific to BV isolates from different macaque species. Restriction enzyme digest patterns of viral DNA revealed marked similarities between rhesus and Japanese macaque isolates, while pigtail and cynomolgus macaque isolates had distinctive cleavage patterns. To further compare genetic diversity among BV isolates, DNA sequences from two regions of the viral genome containing genes that are conserved (UL27 and US6) and variable (US4 and US5) among primate alphaherpesviruses, as well as from two noncoding intergenic regions, were determined. From these sequence data and a phylogenetic analysis of them it was evident that while all isolates were closely related strains of BV, there were three distinct genotypes. The three BV genotypes were directly related to the macaque species of origin and were composed of (i) isolates from rhesus and Japanese macaques, (ii) cynomolgus monkey isolates, and (iii) isolates from pigtail macaques. This study demonstrates the existence of different BV genotypes which are related to the macaque host species and thus provides a molecular basis for the possible existence of BV isolates which vary in their levels of pathogenicity for nonmacaque species.     

A BAC-based contig map of the cynomolgus macaque (Macaca fascicularis) major histocompatibility complex genomic region

The construction of a cynomolgus macaque (Macaca fascicularis, Mafa) BAC library for genomic comparison between rhesus and cynomolgus macaques is necessary to promote the cynomolgus macaque as one of the important experimental animals for future medical and biological research. In this paper, we constructed a cynomolgus macaque BAC library and a map of the MHC (Mafa) genomic region for comparison of the genomic organization and nucleotide similarities between the human, the chimpanzee, and the rhesus macaque. The BAC library consists of 221,184 clones with an average insert size of 83 kb, providing a sixfold coverage of the haploid genome. A total of 114 BAC clones and 54 PCR primer sets were used to construct a 4.3-Mb contig of the MHC region. Diversity analysis of genomic sequence from selected subregions of the MHC revealed that the cynomolgus sequence varied compared to rhesus macaque, human, and chimpanzee sequences by 0.48, 4.15, and 4.10%, respectively. From these findings, we conclude that the BAC library and Mafa genomic map are useful tools for genome analysis and will have important applications for comparative genomics and identifying regions of consequence in medical research.     

Study of cynomolgus monkey (Macaca fascicularis) Mhc DRB gene polymorphism in four populations

The cynomolgus macaque (Macaca fascicularis) is currently used as an animal model in various fields of immunology especially in the development of innovative vaccines for the prevention and treatment of infectious diseases. The polymorphism of the major histocompatibility complex (MHC) influences the development of adaptive immune responses, and it is crucial to characterize the polymorphism of cynomolgus MHC genes. Among all macaque species, the cynomolgus macaque has the most diversified geographical area encompassing continental and insular populations. By the study of a large sample of animals from the Philippines (N = 359), we have characterized 20 DRB haplotypes. The DRB genotyping was performed by denaturing gradient gel electrophoresis (DGGE) sequencing of exon 2 and was confirmed by polymerase chain reaction-sequence-specific oligonucleotide. The DRB and DRA cDNA of 126 animals were characterized by cloning and sequencing. By means of DGGE sequencing, we characterized the polymorphism of genomic DRB exon 2 in three other cynomolgus macaque population samples (Java, Vietnam, and Mauritius), and we discuss about the origin of the founders of the Mauritian and the Filipino cynomolgus macaque populations.     

Composite hyaluronate-type I collagen-fibrin scaffold in the therapy of osteochondral defects in miniature pigs

The potential of novel scaffold containing sodium hyaluronate, type I collagen, and fibrin was investigated in the regeneration of osteochondral defects in miniature pigs. Both autologous chondrocyte-seeded scaffolds and non-seeded scaffolds were implanted into two defects located in the non-weight-bearing zone of the femoral trochlea (defect A was located more distally and medially, defect B was located more proximally and laterally). Control defects were left untreated. Twelve weeks after the operation, the knees were evaluated in vivo using MRI. Six months after the implantation, the defects were analyzed using MRI, histological, and immunohistochemical analysis. In the A defects of chondrocyte-seeded scaffold group, hyaline cartilage and fibrocartilage was formed, containing type II collagen, acidic and neutral glycosaminoglycans while the non-seeded scaffold group was predominantly filled with fibrocartilage. Defects in the control group were predominantly filled with fibrous tissue. Histomorphometric analysis of photomicrographs revealed a significantly higher amount of hyaline cartilage in the cell-seeded scaffold group in A defects than in other groups. Both scaffold groups in A defects showed significantly less fibrous tissue than cell-seeded defects B and the control group. Both histological and MRI analysis proved that the novel composite scaffold has a potential to regenerate osteochondral defects within six months.     

Treatment of craniomaxillofacial fibrous dysplasia: how early and how extensive?

Twenty-eight craniomaxillofacial fibrous dysplasia patients were treated as early as the symptoms occurred. The principles of surgical treatment were based on the zones of involvement: total excision of dysplastic bone of fronto-orbital, zygoma, and upper maxillary origin (zone 1) and bone reconstruction primarily; conservative excision on hair-bearing skull (zone 2), central cranial base (zone 3), and tooth-bearing bones (zone 4); and optic canal decompression on patients with orbital dysplasia and decreasing visual acuity. Patients were followed for 1 to 11 years (average 5.3 years). No recurrence or invasion of the fibrous dysplasia into the grafted bone was seen. One patient had orthognathic maxillary osteotomy on the reconstructed maxilla 6 years after initial reconstruction. Five of 19 patients with alveolar dysplasia had a recurrence and were reshaped. One patient had mandibular sagittal osteotomies to set back the prognathic, fibrous dysplasic mandible after three attempts at conservative shaving. Another patient with mandibular fibrous dysplasia had recurrence with pain and a hemimandibulectomy with successful immediate free vascularized iliac bone graft reconstruction.     

Surgical treatment of nasal obstruction secondary to craniofacial fibrous dysplasia

Craniofacial fibrous dysplasia, a skeletal disorder most often affecting the mandible or maxilla, can cause a variety of complications. Nasal obstruction secondary to this unusual disorder presents a diagnostic and therapeutic challenge. The nasal obstruction in this case is due to expansion of fibrous lesions in adjacent bones as well as direct involvement of the turbinates with fibrous dysplasia. We describe the surgical correction of nasal obstruction in this unique setting and present a brief update of this intriguing disorder.     

Experimental hydroxyapatite cement cranioplasty.

Hydroxyapatite cement is a calcium phosphate-based material that when mixed with water forms a dense paste that sets within 15 minutes and isothermically converts in vivo to a microporous hydroxyapatite implant. This cement was used to reconstruct bilateral 2.5-cm-diameter full-thickness critical-sized parietal skull defects in six cats. One side was reconstructed with 100 percent hydroxyapatite cement, and the other with a mixture of 50 percent hydroxyapatite cement and 50 percent ground autogenous bone by weight. These animals were sacrificed at 6 and 12 months after implantation. Positive and negative controls also were prepared. The anatomic contour of the soft tissue overlying all hydroxyapatite cement implants was well maintained, there were no wound infections or structural failures, and the implants were well tolerated histologically. None of the negative (unreconstructed) control defects was completely filled with repair bone, and all positive (methyl methacrylate) controls demonstrated foreign-body giant-cell formation and fibrous encapsulation of the implants. Examination of decalcified and undecalcified sections revealed progressive but variable replacement of the cement by new bone and soft tissue without a change in the shape or volume of the hydroxyapatite cement-reconstructed areas. New bone comprised 77.3 and 64.7 percent of the tissue replacing the hydroxyapatite cement and hydroxyapatite cement-bone implants, respectively. Replacement of the hydroxyapatite cement implants by new bone is postulated to occur by a combination of osteoconduction and implant resorption. These results indicate that further experimental research leading to the possible application of hydroxyapatite cement for full-thickness calvarial defect reconstruction in humans is warranted.     

Behavioral and adrenocortical responses of male cynomolgus and lion-tailed macaques to social stimulation and group formation

Several recent studies indicate that primate species may differ in behavioral, and in some instances physiological, responses to social manipulations. These differences in response dispositions have been interpreted as being based on temperamental differences among species. This report extends previous comparative studies of macaques by describing behavioral contrasts observed between males of two macaque species, lion-tailed and cynomolgus macaques, in response to social manipulations. Males were exposed to a mirror, then visually exposed to conspecific neighbors in all pairwise combinations, and then formed into conspecific social groups. Urine samples were collected in the pairing and group formation conditions for cortisol assay. The cynomolgus demonstrated more affiliative behavior than the lion-tails in all conditions, while the lion-tails tended to exhibit more aggressive behavior. Following group formation cortisol values showed a decreasing trend in the cynomolgus, but not in the lion-tails. The cynomolgus rapidly adapted to group living and relations between them were primarily affiliative. In contrast, no affiliative behavior was ever observed in the lion-tail group, which appeared to be highly stressed by group living and was eventually disbanded.     

Divergence and diversity of ULBP2 genes in rhesus and cynomolgus macaques

Non-human primates such as rhesus macaque and cynomolgus macaque are important animals for medical research fields and they are classified as Old World monkey, in which genome structure is characterized by gene duplications. In the present study, we investigated polymorphisms in two genes for ULBP2 molecules that are ligands for NKG2D. A total of 15 and 11 ULBP2.1 alleles and 11 and 10 ULBP2.2 alleles were identified in rhesus macaques and cynomolgus macaques, respectively. Nucleotide sequences of exons for extra cellular domain were highly polymorphic and more than 70 % were non-synonymous variations in both ULBP2.1 and ULBP2.2. In addition, phylogenetic analyses revealed that the ULBP2.2 was diverged from a branch of ULBP2.1 along with ULBP2s of higher primates. Moreover, when 3D structural models were constructed for the rhesus ULBP2 molecules, residues at presumed contact sites with NKG2D were polymorphic in ULBP2.1 and ULBP2.2 in the rhesus macaque and cynomolgus macaque, respectively. These observations suggest that amino acid replacements at the interaction sites with NKG2D might shape a specific nature of ULBP2 molecules in the Old World monkeys.     

The ultrastructure of the otolithic membrane and otolith in the juvenile mummichog,Fundulus heteroclitus

The sagitta otolithic membrane of Fundulus heteroclitus consists of two different zones. A structured zone (gelatinous layer), which usually exhibits a reticulated or honeycomb-like architecture, is composed of tightly arranged fibrous material and covers only the sensory region of the macula. The gelatinous layer extends from the otolith surface to the tips of the sensory hairs, and probably functions primarily as a mechanoreceptor. The arrangement of this zone is closely associated with specific overlying structural features of the otolith surface and may also influence the pattern of mineral deposition to some degree. A nonstructured zone (subcupular meshwork) consists of fibers in very loose networks and covers both sensory and nonsensory regions of the macula. Over the sensory region, some of this fibrous material extends from the epithelial surface, through pores in the gelatinous layer, to the surface of the overlying otolith. In the nonsensory region, fibers of the subcupular meshwork are relatively more numerous and extend around the peripheral margin of the otolith. Evidence is presented which suggests that the fibrous material of the subcupular meshwork is incorporated into the otolith as an organic matrix constituent. New aspects on the ultrastructure of the otolith are presented and discussed.     

Divergence population genetic analysis of hybridization between rhesus and cynomolgus macaques

The geographic ranges of rhesus ( Macaca mulatta ) and cynomolgus ( M. fascicularis ) macaques adjoin in Indochina where they appear to hybridize. We used published and newly generated DNA sequences from 19 loci spanning ~20 kb to test whether introgression has occurred between these macaque species. We studied introgression at the level of nuclear DNA and distinguished between incomplete lineage sorting of ancestral polymorphisms or interspecific gene flow. We implemented a divergence population genetics approach by fitting our data to an isolation model implemented in the software IMa. The model that posits no gene flow from the rhesus into the cynomolgus macaque was rejected ( P  = 1.99 × 10⁻⁸). Gene flow in this direction was estimated as 2 Nm ~1.2, while gene flow in the reverse direction was nonsignificantly different from zero ( P  = 0.16). The divergence time between species was estimated as ~1.3 million years. Balancing selection, a special case of incomplete sorting, was taken into consideration, as well as potential crossbreeding in captivity. Parameter estimates varied between analyses of subsets of data, although we still rejected isolation models. Geographic sampling of the data, where samples of cynomolgus macaques derived from Indochina were excluded, revealed a lost signature of gene flow, indicating that interspecific gene flow is restricted to mainland Indochina. Our results, in conjunction with those by others, justify future detailed analyses into the genetics of reproductive barriers and reticulate evolution in these two genome-enabled primates. Future studies of the natural hybridization between rhesus and cynomolgus macaques would expand the repertoire of systems available for speciation studies in primates.     

X-ray manifestations of fibrous dysplasia of the facial bones]

The paper is concerned with roentgenological characterization of mono- and poly-osseous fibrous dysplasia in cranial facial bones of 60 persons of both sexes and at a different age, of them 14 persons were under 14. Mono-osseous lesions after radical operations were shown to be capable of continued growth of rearranged osseous tissue in those cases when the periosteum was not completely removed.