Cloning and sequencing of the bottle-nosed dolphin (Tursiops truncatus) interferon-gamma gene.

The bottle-nosed dolphin (Tursiops truncatus) interferon-gamma(IFN-gamma) cDNA was cloned from mitogen stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The dolphin IFN-gammacDNA was 585 base pairs in length and contained an open reading frame of 498 nucleotides encoding an IFN-gammaprecursor of 166 amino acids, with cleavage of the putative signal peptide of 19 amino acids. Analysis of the mature amino acid sequences revealed two potential N-linked glycosylation sites. Comparison of the predicted amino acid sequence showed that dolphin IFN-(gamma)shares 86, 85, 63 and 42% similarity with the bovine, ovine, human and mouse IFN-gamma, respectively.

Cloning and sequencing of a bottle-nosed dolphin (Tursiops truncatus) interleukin-4-encoding cDNA.

Using polymerase chain reaction (PCR), a bottle-nosed dolphin (Tursiops truncatus) interleukin-4 (IL4) cDNA was cloned and sequenced. IL4 specific primers were based on the 5' and 3' untranslated regions of the human and murine IL4 gene. The dolphin IL4 cDNA is 528 base pairs in length and contains an open reading frame of 402 nucleotides coding an IL4 precursor of 133 amino acids, with the putative signal peptide of 24 amino acids. Analysis of the mature amino acid sequence shows three potential N-linked glycosylation sites and three disulfide bonds. Comparison of the predicted amino acid sequence shows that dolphin IL4 shares 77, 74, 58 and 41% identity with the bovine, ovine, human and mouse IL4s, respectively.

Cloning and sequencing of a bottle-nosed dolphin (Tursiops truncatus) interleukin-1alpha and -1beta complementary DNAs.

The bottle-nosed dolphin (Tursiops truncatus) IL-1alpha and IL-1beta cDNA were cloned from mitogen stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The sequences of these cDNAs showed that dolphin IL-1alpha and IL-1beta clones contained open reading frames encoding 265 and 266 amino acids, respectively. Comparison of the deduced amino acid showed that dolphin IL-1alpha sequence shared 77, 77, 77, 74, 71, 65 and 57% similarity with the bovine, ovine, porcine, equine, feline, human and mouse IL-1alpha sequences, respectively. Similarly, the amino acid sequence showed that dolphin IL-1beta shared 77, 77, 74, 69, 65, 64 and 63% similarity with the bovine, ovine, porcine, equine, feline, human and mouse IL-1beta sequences, respectively. The relatedness of dolphin IL-1alpha and IL-1beta were relatively distant with 21% amino acid homology.

Molecular cloning of bottle-nosed dolphin (Tursiops truncatus) MHC class I cDNA.

Using polymerase chain reaction, bottle-nosed dolphin (Tursiops truncatus) major histocompatibility complex (DoLA) class I alpha chain cDNA was cloned and sequenced. Predicted amino acid sequences of DoLA class I alpha chain have cystein residues for intradomain disulfide bond formation and N-linked glycosylation sites, suggesting that DoLA class I alpha chain molecules construct alpha 1, alpha 2, and alpha 3 domain structures. Similarity of DoLA class I alpha chain cDNA to land mammal MHC class I alpha chain cDNA was 90.4% in cattle, 90.2% in horse, 89.4% in sheep, and 87.8% in human. This investigation suggests that DoLA is closely related to land mammals.

A monoclonal antibody against dolphin lymphocytes (6E9) which recognizes bovine MHC class II antigens.

A monoclonal antibody, 6E9 established from mice injected with dolphin peripheral blood lymphocytes (PBLs) was characterized. In addition to its reactivity against 89.4% of dolphin PBLs, 6E9 reacted with 33.1% of bovine PBLs of which 22% were CD5+, 11.1% were CD5-. 6E9 recognized a 34 kD protein on the surface of dolphin and bovine PBLs. Analysis of the protein's N-terminal amino acid sequence indicated that 6E9 recognizes bovine major histocompatibility complex (MHC) class II antigens. These results suggested that 6E9 recognized MHC class II antigens on bovine PBLs. As we have already produced an anti-dolphin MHC class I monoclonal antibody, analysis of immune system using these monoclonal antibodies will advance our understanding of the evolution of the mammalian immune system.

[Radiological study of the accessory skeletal elements in the foot and ankle (author's transl)].

The accessory bones and ossification centers in the foot and ankle were studied in 3,460 radiographs of patients over 7 years of age who consulted our clinic for various reasons from January 1967 to December 1979. The patients comprised of 1,639 males and 1,821 females. The accessory ossicles commonly observed were os tibiale externum, os trigonum, and os peroneum in order of frequency. The os tibiale externum is the commonest accessory bones in the foot and is situated at the point where the tendon of the posterior tibial muscle inserts itself into navicular bone. This ossicle was found to be present in 733 out of 3,460 feet (21.3%). As some authors have pointed out, they are observed somewhat more frequently in females than in males. The os trigonum is an accessory bone which is situated behind the posterior process of the talus. This ossicle was found in 438 feet (12.7%). The os peroneum is a sesamoid in the long peroneal tendon where it bends toward the sole of the foot. This was found in 311 feet (9.0%). Other inconstant skeletal elements were less commonly observed. This os intermetatarseum which is usually located in the first, rarely in the fourth, intermetatarsal space, was found in 91 feet (2.6%). When studied meticulously, this ossicle is found more often than is generally believed. The os subfibulare, situated just under the lateral malleolus, was found in 71 feet (2.1%). The os supranaviculare, situated above the talonavicular joint, was found in 35 feet (1.0%). The os subtibiale, situated just under the medial malleolus, was found in 31 feet (0.9%). The os supratalare, situated above the neck of the talus, was found in 25 feet (0.9%). The calcaneus secundarius, surrounded by calcaneus, talus, os naviculare and os cuboideum, was found in 20 feet (0.6%). The os talotibiale, situated in front of the talocrural joint, was found in 16 feet (0.5%). The os sustentaculi, situated beside the sustentaculum tali, was found in 9 feet (0.3%). The talus accessorius, situated beside the trochlea tali, was found in 6 feet (0.2%). The os vesalianum, situated adjacent to the tuberositas ossis metatarsalis V, was found in 4 feet of the adult patients (0.1%). The talus secundarius, situated near the trochlea peronealis calcanei, was found in 3 feet (0.1%). The os intercuneiforme, located between first and second cuneiform and the navicular bone, was found in only one foot of a 33-year-old male (0.03%). This case may be the first observation in Japan of this extremely rare accessory ossicle. The majority of the patients who had these accessory bones in their feet and ankles seemed to have no clinical complaints. Identification of these ossicles is of importance because they may easily be mistaken as fragments of avulsion fractures. An extra center of ossification for the medical malleolus was observed in 11 feet of the children aged 8 to 11 years. The ossification center for the tuberositas ossis metatarsalis V was found in 56 feet of the adolescents...