A novel nonsense mutation in the tyrosinase gene is related to the albinism in a capuchin monkey (Sapajus apella).

BACKGROUND: Oculocutaneous Albinism (OCA) is an autosomal recessive inherited condition that affects the pigmentation of eyes, hair and skin. The OCA phenotype may be caused by mutations in the tyrosinase gene (TYR), which expresses the tyrosinase enzyme and has an important role in the synthesis of melanin pigment. The aim of this study was to identify the genetic mutation responsible for the albinism in a captive capuchin monkey, and to describe the TYR gene of normal phenotype individuals. In addition, we identified the subject's species. RESULTS: A homozygous nonsense mutation was identified in exon 1 of the TYR gene, with the substitution of a cytosine for a thymine nucleotide (C64T) at codon 22, leading to a premature stop codon (R22X) in the albino robust capuchin monkey. The albino and five non-albino robust capuchin monkeys were identified as Sapajus apella, based on phylogenetic analyses, pelage pattern and geographic provenance. One individual was identified as S. macrocephalus. CONCLUSION: We conclude that the point mutation C64T in the TYR gene is responsible for the OCA1 albino phenotype in the capuchin monkey, classified as Sapajus apella.

Rotaviruses reach late endosomes and require the cation-dependent mannose-6-phosphate receptor and the activity of cathepsin proteases to enter the cell.

UNLABELLED: Rotaviruses (RVs) enter cells through different endocytic pathways. Bovine rotavirus (BRV) UK uses clathrin-mediated endocytosis, while rhesus rotavirus (RRV) employs an endocytic process independent of clathrin and caveolin. Given the differences in the cell internalization pathway used by these viruses, we tested if the intracellular trafficking of BRV UK was the same as that of RRV, which is known to reach maturing endosomes (MEs) to infect the cell. We found that BRV UK also reaches MEs, since its infectivity depends on the function of Rab5, the endosomal sorting complex required for transport (ESCRT), and the formation of endosomal intraluminal vesicles (ILVs). However, unlike RRV, the infectivity of BRV UK was inhibited by knocking down the expression of Rab7, indicating that it has to traffic to late endosomes (LEs) to infect the cell. The requirement for Rab7 was also shared by other RV strains of human and porcine origin. Of interest, most RV strains that reach LEs were also found to depend on the activities of Rab9, the cation-dependent mannose-6-phosphate receptor (CD-M6PR), and cathepsins B, L, and S, suggesting that cellular factors from the trans-Golgi network (TGN) need to be transported by the CD-M6PR to LEs to facilitate RV cell infection. Furthermore, using a collection of UK × RRV reassortant viruses, we found that the dependence of BRV UK on Rab7, Rab9, and CD-M6PR is associated with the spike protein VP4. These findings illustrate the elaborate pathway of RV entry and reveal a new process (Rab9/CD-M6PR/cathepsins) that could be targeted for drug intervention. IMPORTANCE: Rotavirus is an important etiological agent of severe gastroenteritis in children. In most instances, viruses enter cells through an endocytic pathway that delivers the viral particle to vesicular organelles known as early endosomes (EEs). Some viruses reach the cytoplasm from EEs, where they start to replicate their genome. However, other viruses go deeper into the cell, trafficking from EEs to late endosomes (LEs) to disassemble and reach the cytoplasm. In this work, we show that most RV strains have to traffic to LEs, and the transport of endolysosomal proteases from the Golgi complex to LEs, mediated by the mannose-6-phosphate receptor, is necessary for the virus to exit the vesicular compartment and efficiently start viral replication. We also show that this deep journey into the cell is associated with the virus spike protein VP4. These findings illustrate the elaborate pathway of RV entry that could be used for drug intervention.

Elucidation of the time course of adenosine deaminase APOBEC3G and viral infectivity factor vif in HIV-2(287) -infected infant macaques.

BACKGROUND: Although the interactions of cellular cytidine deaminase A3G and viral infection factor (vif) of human immunodeficiency virus (HIV) were reported, regulation of A3G after in vivo HIV infection and disease progression is not known. METHODS: Time courses of plasma virus, CD4(+) T lymphocyte Macaca levels, and concentrations of A3G and vif transcripts were determined in infant macaques infected with HIV-2(287) . These in vivo results were compared with those collected in vitro in HIV-2-infected T cells. RESULTS: Human immunodeficiency virus-infected macaques exhibited plasma viremia (≥10(8) copies/ml) followed by a precipitous CD4(+) T-cell (from 40-70 to ≤5%) decline. An initial increase in A3G transcripts coincides with early increases in virus and vif RNA. As virus load continues to increase, A3G RNA decreases but recovers at a later phase as virus level stabilizes. Pearson correlation analysis revealed strong interactions of A3G-CD4, vif-CD4, and A3G-vif. CONCLUSIONS: There is a time-dependent A3G and vif RNA interaction throughout the course of HIV infection.

Erythrocyte characteristics in vitamin E-responsive anemia of the owl monkey (Aotus trivirgatus).

To characterize the vitamin E-responsive anemia occurring in owl monkeys (Aotus trivirgatus), osmotic fragility, and H2O2-induced and time-dependent hemolysis, as well as RBC lipid peroxidation, were compared in anemic and nonanemic owl monkeys. Whereas vitamin E serves as a lipid-soluble antioxidant, the glutathione peroxidase system functions in the water-soluble phase of the cell. Thus, activity of glutathione peroxidase, glutathione reductase, and glucose-6-phosphate dehydrogenase, as well as reduced glutathione concentrations in owl monkeys' RBC, were compared with those of rhesus macaques and cebus and squirrel monkeys fed the same diet and maintained under the same management scheme. Osmotic fragility did not differ between anemic and nonanemic owl monkeys. The H2O2-induced and time-dependent hemolysis was approximately 10-fold greater among anemia owl monkeys than among their nonanemic counterparts, and lipid peroxidation values tended to be higher in the anemic monkeys. Owl monkeys, as a species and independent of anemia, exhibited higher RBC peroxidation than did 2 other New World species, cebus and squirrel monkeys. The glutathione peroxidase system was not depressed in owl monkey RBC. The only observed difference in this system was in the glucose-6-phosphate dehydrogenase activity, which was 3- to 6-fold higher in the owl monkey than in the other species, indicating an increased activity of the peroxidase system. Thus, a defect in the glutathione peroxidase system could not be identified.

[Epizootic hepatitis A among African green monkeys kept in a vivarium]. / Epizootiia gepatita A sredi afrikanskikh zelenykh martyshek, soderzhashchikhsia v usloviiakh vivariia.

The results of observations on the pattern of spread of hepatitis A virus and immune response to it in African green monkeys (Cercopithecus aethiops) kept in the animal house are presented. The infection in the monkeys was found to be characterized by all virological, serological, and biochemical parameters inherent in hepatitis A virus. The results indicated that hepatitis A in monkeys may run both asymptomatic and clinically manifest course, and the spread of infection in the animal house sequentially involves most seronegative animals into the epidemic process.

[Malignant non-Hodgkin's lymphomas forming from the germinal-center cells of the lymphoid follicles in baboons from a high-risk stock]. / Nekhodzhkinskie zlokachestvennye limfomy, formiruiushchiesia iz kletok zarodyshevykh tsentrov limfoidnykh follikulov u pavianov stada povyshennogo riska.

Histological, cytochemical and ultrastructural investigation of immunologically typed B-cell non-Hodgkin's malignant lymphomas (NHL) of primates (model system on baboons) revealed 15 cases of malignant lymphomas originating from germinal centre cells of lymph nodes follicles. By the tumour cell type centroblastic (CB), centroblastic/centrocytic (CB/CC) and centrocytic (CB), malignant lymphomas were distinguished (according to Kiel classification). In case of CB NHL, tumours, as a rule, are of nodular type. Tumours, in which centrocytic infiltration predominates, are characterized by diffuse type of growth in lymphoid and nonlymphoid organs. Generalized process affects mainly lymph nodes and to considerably lower degree involves spleen and nonlymphoid parenchymatous organs.

[Serum creatine phosphokinase, GOT, and GPT activity in different types of infectious diseases in monkeys]. / Aktivnost' kreatinfosfokinazy, AST and ALT v syvorotke krovi pri nekotorykh vidakh infektsionnoi patologii obez'ian.

An increase in creatine phosphokinase (CPK) activity as well as a decrease in activities of aspartate aminotransferase (AAT) and alanine aminotransferase (ALT) were observed in monkey blood serum under conditions of experimental infection of the animals with dysentery. Contrary to the non-immunized animals, the activity of CPK was increased but the activities of AAT and ALT were unaltered in blood serum of the immunized monkeys. Estimation of the enzymatic activity might be used for diagnosis of experimental dysentery and streptococcal infection.

Relationship of creatine kinase, aspartate aminotransferase, lactate dehydrogenase, and proteinuria to cardiomyopathy in the owl monkey (Aotus vociferans).

BACKGROUND: The purpose of this study was to determine serum reference values for creatine kinase (CK), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in captive-born and wild-caught owl monkeys to assess their usefulness for diagnosing myocardial disease. METHODS: Blood samples were collected for CK, AST and LDH determinations. In addition, urine samples were collected and semiquantitative tests performed. RESULTS: There was no statistically significant difference between CK, AST and LDH when comparing both groups. However, when comparing monkeys with proteinuria to those without proteinuria, a statistically significant difference in CK value was observed (P = 0.021). In addition the CK/AST ratio revealed that 29% of the animals included in this study had values suggesting cardiac infarction. Grossly, cardiac concentric hypertrophy of the left ventricle and small, pitted kidneys were the most common findings. Microscopically, myocardial fibrosis, contraction band necrosis, hypertrophy and hyperplasia of coronary arteries, medium-sized renal arteries and afferent glomerular arteriolae were the most significant lesions, along with increased mesangial matrix and hypercellularity of glomeruli, Bowman's capsule and peritubular space fibroplasia. CONCLUSIONS: These findings suggest that CK, AST and LDH along with urinalysis provide a reliable method for diagnosing cardiomyopathies in the owl monkey. In addition, CK/AST ratio, proteinuria and the observed histological and ultrastructural changes suggest that Aotus vociferans suffer from arterial hypertension and chronic myocardial infarction.

Evidence of hepatitis A infection in immature rhesus monkeys.

Forty immature (less than 2 years old) rhesus monkeys (Macaca mulatta) with marked increases in aspartate and alanine aminotransferase activities were examined. Serological and histopathological evaluations were done to determine if affected animals were infected with hepatitis A virus. Although no clinical signs of illness were noted in any of the monkeys, an excellent correlation was found between the increased serum aminotransferase values and seropositivity with the acute phase (IgM) HAVAB-M antibody. Histopathological evaluations of livers of selected animals revealed hepatic lesions consistent with those in chimpanzees and marmosets infected with hepatitis A virus: generalized activation of sinusoidal lining cells, focal hepatocellular necrosis with occasional acidophilic bodies, and cuffs of mononuclear cells in the portal areas. Although no animals were seropositive for HAVAB upon receipt from the breeding colony, a total of ten of 18 animals for which serological data were available had seroconverted to HAVAB positivity during the 4-month observation period. These results are consistent with hepatitis A infection in immature rhesus monkeys and indicate the potential significance of serological testing in animals in which hepatic function is being evaluated.

Serum lactate dehydrogenase of normal, stressed, and yellow fever virus-infected rhesus monkeys.

Serum lactate dehydrogenase enzyme (sLDH) was studied in both healthy and yellow fever virus (YFV)-infected young adult rhesus monkeys (Macaca mulatta). In healthy monkeys, significant variation (p less than 0.001) was observed for both total activity and isoenzyme distribution among the following comparisons: individual monkeys, different days, different times of day, and caged versus chair-restrained monkeys (until 7 da after chair restraint). However, variability of baseline values was reduced by the use of samples obtained from resting subjects at 9:00 am on at least 3 consecutive days. Normal total activity and isoenzyme distribution values were based on 148 determinations obtained from 73 healthy, caged monkeys. Both total activity and the proportion of the 5th isoenzyme fraction increased significantly (p less than 0.001) in the YFV-infected monkeys, beginning 90 hr postinfection, consistent with hepatocellular necrosis and release of isoenzymes into the serum. The assay for sLDH appears to be of value as a diagnostic indicator during the course of YFV infection in the rhesus monkey.

Characterization of the rhesus monkey galactocerebrosidase (GALC) cDNA and gene and identification of the mutation causing globoid cell leukodystrophy (Krabbe disease) in this primate.

Krabbe disease or globoid cell leukodystrophy (GLD) is a severe lysosomal disorder resulting from the deficiency of galactocerebrosidase (GALC) activity. This deficiency results in the insufficient catabolism of several galactolipids that are important in the production of normal myelin. Since the cloning of the human GALC cDNA and gene many disease-causing and polymorphic changes have been identified. This autosomal recessive disease has been reported to occur in several animal species, and recently the murine and canine GALC genes have been cloned. We now describe the cloning of the GALC cDNA and gene from the rhesus monkey and the identification of the mutation causing GLD in this species. The nucleotide sequence of the coding region and the gene organization were nearly identical to human. The deduced amino acid sequence of the monkey GALC was compared to the human, dog, and mouse, and it was found to be 97, 87, and 83% identical, respectively. The mutation causing GLD in the rhesus monkey is a deletion of AC corresponding to cDNA positions 387 and 388 in exon 4. This results in a frame shift and a stop codon after 46 nucleotides. A rapid method to detect this mutation was developed, and when 45 monkeys from this colony were tested, 22 were found to be carriers. The availability of this nonhuman primate model of GLD will provide unique opportunities to evaluate treatment for this severe disease.

Cyclooxygenase-2 expression in inflammatory lung lesions of nonhuman primates.

Mammalian cells contain two related but unique isoforms of cyclooxygenase (COX-1 and COX-2). COX-1 is expressed constitutively in a majority of tissues and is involved in the production of prostaglandins (PGs) that modulate normal physiologic functions. COX-2 is inducible by various stimuli and is involved in the production of PGs that modulate physiologic events in development, cell growth, and inflammation. With the exception of peribronchial glands and chondrocytes of peribronchial cartilage, COX-2 is not detectable in the normal lung of nonhuman primates. We evaluated COX-2 expression by immunohistochemical methods in the inflammatory lesions of two cynomolgus monkeys (Macaca fascicularis) with acute severe pneumonia. Both monkeys exhibited acute severe bronchopneumonia; histologically, lung lesions were characterized by infiltration of large numbers of neutrophils and fewer macrophages, mild bronchial epithelial hyperplasia, and slight type-2 pneumocyte hyperplasia. In both monkeys, mild to marked COX-2 immunoreactivity was detected within the cytoplasm of macrophages, bronchial epithelial cells, type-2 pneumocytes, and endothelial cells of blood vessels. No COX-2 immunoreactivity was detectable in the neutrophils that constituted >90% of the inflammatory cells. These observations suggest that in acute inflammatory lung lesions in nonhuman primates 1) COX-2 is induced in the bronchial and alveolar epithelial cells, 2) macrophages are the primary inflammatory cells that exhibit COX-2, and 3) neutrophils do not express COX-2.

Genetic galactocerebrosidase deficiency (globoid cell leukodystrophy, Krabbe disease) in rhesus monkeys (Macaca mulatta).

Globoid cell leukodystrophy, or Krabbe disease, is a severe disorder of the peripheral and central nervous system myelin caused by deficient galactocerebrosidase (GALC) activity. This autosomal recessive disease affects humans and animals including dogs, mice, and rhesus monkeys. Cloning of the human and animal GALC genes opened opportunities for therapeutic trials using animal models. We describe the clinical, pathologic, and biochemical features of the affected rhesus monkey. Affected monkeys had very low GALC activity and a two base pair deletion in both copies of the GALC gene. Clinical signs of tremors, hypertonia, and incoordination led to humane euthanasia by 5 months of age. At necropsy, peripheral nerves were enlarged. Microscopically, the cerebral, cerebellar, and spinal cord white matter was infiltrated with periodic acid-Schiff-positive multinucleated globoid cells, and there was a striking lack of myelin. Peripheral nerve fibers were decreased in number and separated by Alcian blue- and safranin O-positive material. Myelin sheaths were greatly diminished. Lipid analysis of brains of 12-day-old and 158-day-old affected monkeys revealed a great excess of psychosine in white matter. The rhesus monkey model will be especially useful for exploring treatment options, including prenatal bone marrow transplantation and various approaches to gene therapy.