Koshi Yamamoto

Isolation of Mycoplasma pneumoniae from Pleural Fluid and/or Cerebrospinal Fluid: Report of Four Cases

Mycoplasma pneumoniae was isolated from pleural exudates of 2 children with pleuropneumonia. The organism was also isolated from cerebrospinal fluid of a child with meningeal manifestations and from the cerebrospinal fluid as well as pleural exudate of another child who died 21 days after onset of disease. More extensive attempts to cultivate M. pneumoniae from nonpulmonary sites are recommended.

Identification of the sequence responsible for the nuclear localization of human Cdc6

The Cdc6 is the essential protein for the initiation of DNA replication. Cdc6 is localized in the G1 nucleus, and abnormal nuclear localization of this protein induces irregular initiation of DNA replication. We identified here that amino acids K57 and R58 in the human Cdc6 protein play an important role in the nuclear localization of the protein. The fundamental features of the mechanism regulating the localization of Cdc6 seem to be maintained in yeast, Xenopus, and human, since the amino acid sequence surrounding K57 and R58, (S/T)PXKR(L/I), is conserved in these species. Substitution of amino acid residue S54 with E and not Q blocked partially the nuclear localization of the protein, implying that the phosphorylation at S54 is involved in the regulating mechanism of the cell cycle‐dependent localization of Cdc6.

Mycoplasma pneumoniae and other pathogens in the aetiology of lower respiratory tract infections among Japanese children.

The causes of lower respiratory tract infections in 1544 children attending a Japanese hospital over a period of 7 years were investigated. Both cultivation and two serological techniques were used to detect Mycoplasma pneumoniae whereas viral involvement was investigated only by serology. Pathogens were identified in 52% of 1175 patients with pneumonia and 36% of 369 patients without pneumonia. Mycoplasma pneumoniae infection was specifically diagnosed in 414 (26.8%) of the 1544 patients. Respiratory syncytial virus (RSV) was dominant in children up to 2 years of age. By the age of 3 years, M. pneumoniae was equalling it in incidence and became the main pathogen in older groups. It is suggested that M. pneumoniae may be more important in the 3-6 years age group than hitherto suspected. These observations may influence the choice of antibiotics for treating lower respiratory tract infections in childhood.

Isolation of T-mycoplasmas from cats in Japan.

The first isolation of T-mycoplasmas from the throat swabs of clinically healthy cats was described by Tan and Markham in New Zealand in 1971 (5). Tan and Miles further implied the possible role of feline T-mycoplasmas in cat abortions (6). No other reports, however, have been published concerning feline T-mycoplasma. To date, no relationship between T-mycoplasmas originating from humans and cats has been found. This communication reports the first isolation of T-mycoplasma from cats in Japan and serological comparisons, using the immune sera raised against eight serotypes of human T-mycoplasmas. Media for isolation and growth of T-mycoplasmas were as previously reported (7). Thirty-six cats were examined for the presence of T-mycoplasmas. Group I included 26, short-haired, laboratory cats which were apparently healthy. Group II consisted of ten house cats, including not only short-haired but also Siamease and Persian cats, which were brought to a veterinary hospital. The age of the cats, ranging from two months to five years, was not controlled in the present study. Using sterile cotton-tipped swabs, specimens were collected from the oral cavities and vaginas or prepuces of the cats. The swabs were immediately immersed in the liquid medium and then incubated at 30 C. When a color change was observed in the liquid cultures, drops of the culture were placed on the solid medium plates and incubated at 30 C in a humidified chamber with about 10% carbon dioxide. The isolates were cloned by transferring single colonies from the solid medium into the liquid medium. After incubation at 30 C, the broth culture which changed in color was filtered through a 450 nm membrane filter and the dilutions of the filtrate were placed on the solid medium to produce well-isolated single colonies. The whole procedure was repeated twice and, after the third single colony picking, the subcultures were maintained as stock strains for further characterization. As shown in Table 1, T-mycoplasmas were isolated from 25 out of 36 cats. The frequencies of isolation of T-mycoplasmafrom the oral cavities and vaginas or pre-

Serological Studies with Feline Ureaplasmas

ganisms only from the same site, suggesting that this region was the preferred site of colonization (10). As described by Harasawa et al (1), however, the organisms have also been isolated from the oral cavity and, less frequently, from the urogenital tract of cats. Tan and Miles (12) have suggested the possible role of the organisms in cat abortions with an experimental inoculation of a strain isolated from a feline throat into pregnant dams. No antigenical relationship between feline and human ureaplasmas in growth inhibition (11) or metabolism inhibition (1) tests has been detected. In addition, Howard et al (3) have reported that the guanine-plus-cytosine (G+ C) content of DNA of a feline strain was similar to canine and human strains, but different from bovine, caprine, ovine, and simian strains. No report has been published concerning serological typing of feline ureaplasmas. This report describes the results of serological studies with feline ureaplasmas. The media used for growth and serology of ureaplasmas were as described previously (8). Eleven feline ureaplasma strains were cloned by picking single colonies as previously noted (8). Among them, nine strains, i.e. FT1-B, FT2-B, FT2-C, FT3-A, FT3-B, FT5-A, FT5-C, FT10-A, and FT10-C, were isolated from the oral cavity of cats at this laboratory as reported before (1), and two strains, F45d2 and F2, isolated from the same site were provided by Dr. K. Koshimizu, of the Faculty of Medicine, University of Tokyo. Eight human strains, serotypes I to VIII (9), seven bovine strains, i.e. C13, C24, C23, C17, C12, C7, and T44 (8), four canine strains, i.e. D1M-C, D29M, D11N-A, and D6P-C (6), a simian strain T167-2 (4), and an avian strain D6-1 (5) were employed as reference strains for comparison. Eleven feline strains were tested by the growth inhibition test (8) with four antisera (Table 1). Ten strains, FT1-B to F45d2, were inhibited in growth by the antisera to strains FT2-B, FT2-C, and F45d2, and were apparently identical. However, none of these strains were inhibited by the antiserum to strain F2. The remaining strain F2 reacted with only the homologous antiserum, and was thus distinct

Inhalation anesthesia with GOF in woodchucks.

Woodchucks (Marmota monax) with cysticercosis or with liver cancer induced by WHV were successf my operated under GOF anesthesia.Premedication was carried out with atropine sulfate (0.05 mg/kg, IM), acetylpromazine maleate (0.05 or 0.10 mg/kg, IM) and ketamine hydrochloride (20 mg/kg, IM) . Approximately 15 min after the premedication, the animals were well sedated and easily and safely restrained.After mask in halation of GOF, they were intubated and maintained with 1-1.5% halothane (O2: N2O=1: 1) . During the surgery, changes in heart rate, respiratory rate and body temperature were not significant, except in the woodchuck with cysticercosis, in which dopamine was infused during the rest of the surgery.Recovery from the anesthesia was very fast in all the animals except the one with cysticercosis and tracheal tubes were removed in approximately 10 min after the end of the halothane inhalation.These results suggest that GOF anesthesia with this type of premedication is safe and practical in the surgery of woodchucks.