Genomic comparison of adenovirus type 3 isolates from patients with acute conjunctivitis in Japan, Australia, and the Philippines.

A total of 53 Ad3 isolates obtained from patients with acute conjunctivitis in Japan, Australia, and the Philippines during 1973 to 1986 were analyzed for their genome types with 4 restriction endonucleases, BamHI, BglII, HindIII, and SmaI. Two new genome types designated tentatively as Ad3f and Ad3g were identified by combination of BamHI and BglII in the isolates. The changes of restriction sites and sizes of restriction fragments in newly recognized Ad3f and Ad3g were located at the similar regions reported in other Ad3 genome types by O'Donnell et al (1986) on physical maps of the Ad3 prototype strain GB genome.

Biological screening of monoclonal antibodies.

A method was developed to screen hybridomas secreting immunoglobulin to cell surface receptors by observing the ability of antibodies to inhibit cell attachment and survival. The model used to develop the screening procedure involved mouse hybridomas secreting monoclonal IgG to human epidermal growth factor (EGF) receptors. Conditioned medium from these hybridomas inhibited the attachment and subsequent growth of human foreskin fibroblasts unless excess EGF was added to the cultures.

Quantitative autoradiographic analysis of muscarinic cholinergic and adenosine A1 binding sites after transient forebrain ischemia in the gerbil.

The influence of transient forebrain ischemia on adenosine A1 and muscarinic cholinergic receptors in the gerbil brain 1-27 days after recirculation was studied. The topographical distribution and the alteration in the adenosine A1 and muscarinic receptor sites were analyzed by means of quantitative receptor autoradiography using [3H]cyclohexyladenosine ([3H]CHA) and [3H]quinuclidinyl benzilate ([3H]QNB), respectively. In most regions examined, the temporal profiles of the alteration of the receptor density were in accordance with the histopathological findings.

GABA and benzodiazepine receptors in the gerbil brain after transient ischemia: demonstration by quantitative receptor autoradiography.

Quantitative receptor autoradiography was used to measure the binding of gamma-aminobutyric acid (GABA) and benzodiazepine receptors after ischemia by means of transient occlusion of bilateral common carotid arteries in the gerbil. [3H]Muscimol was used to label the GABAA receptors and [3H]flunitrazepam to label central type benzodiazepine receptors. In the superolateral convexities of the frontal cortices, [3H]muscimol binding was increased in 60% of the animals killed 3 days after ischemia, and decreased in 67% of the animals killed 27 days after ischemia.

Phylogenetic relationships between adenoviruses as inferred from nucleotide sequences of inverted terminal repeats.

The nucleotide (nt) sequences of inverted terminal repeats (ITR) from human adenovirus (Ad) 19, bovine Ad1 (BAd1), bovine Ad3 (BAd3), canine Ad2 (CAd2) and an avian Ad, EDS-76, were determined. The length of the ITR sequence was 160 bp in Ad19, 159 bp in BAd1, 195 bp in BAd3, 196 bp in CAd2 and 52 bp in EDS-76. CAd2 had the longest ITR among the examined Ads, BAd3 the second longest, and EDS-76 had the shortest ITR.

Immunoreactivity of a synthetic pentadecapeptide corresponding to the N-terminal region of the scrapie prion protein.

A pentadecapeptide with an amino acid sequence corresponding to the amino-terminal region of the scrapie prion protein was synthesized. Immunization of a rabbit with the peptide conjugated with ovalbumin induced specific antibodies. The antibodies reacted with all three of the major polypeptides in a proteinase K-treated fraction obtained from brains of mice infected with the Obihiro strain of scrapie agent.

Lesions to Schaffer collaterals prevent ischemic death of CA1 pyramidal cells.

The contribution of excitatory inputs to CA1 pyramidal cell death after ischemia was examined using rats with unilateral destruction of CA3 pyramidal cells. Intracerebroventricular injection of L-alpha-kainic acid (KA) was performed before the induction of transient forebrain ischemia. Five days after ischemic insult, pyramidal cells and L-glutamate binding sites in the CA1 region ipsilateral to the KA injection were preserved in spite of neuronal necrosis and a significant decrease in L-glutamate receptor density in the contralateral CA1 region, indicating the critical role of Schaffer collaterals in delayed neuronal death.

Real-time monitoring of the secretory function of cultured adrenal chromaffin cells.

A system to discriminate the real-time dynamics of the secretory function in cultured adrenal chromaffin cells, using a cell bed perfusion technique and an amperometric detector, was established. Examination of basal conditions revealed that the electrode potential and flow rate are crucial factors for monitoring precise dynamics of the secretory process. Stimulation of the cells either with acetylcholine (ACh) or with high K+ concentration caused a transient current response.

Developing differentiated epithelial cell cultures: airway epithelial cells.

Recent progress in cell culture enables us to grow and to maintain differentiated epithelial cells in a serum-free defined culture environment. Such an epithelial cell culture system free from interference by other nonepithelial cell types should be used widely in studies related to toxicology, carcinogenesis, and disease-related problems. This approach will lead to a better understanding of pathological changes indicated in the injured epithelial layer.

Generation of packaging-defective DNA molecules of equine adenovirus.

Equine adenovirus (EAd) DNA prepared from infected bovine kidney (MDBK) cells contained additional sequences of about 100 to 700 bp at the left-hand end of the genome. These aberrant viral genomes were produced even after the first passage of the wild type EAd in MDBK cells and their relative amounts did not change significantly during serial passage. The left terminal fragments of two defective viral DNAs were cloned into the plasmid vector pBR322 and the nucleotide sequences of their terminal regions were analyzed.

Biochemical characterization of the cholesterol-dependent growth of the NS-1 mouse myeloma cell line.

The biochemical basis for the cholesterol-dependent growth phenotype of the NS-1 myeloma cell line has been investigated. In one series of experiments, the growth response of NS-1 cells to several of the intermediates of cholesterol biosynthesis was studied in serum-free medium. The cholesterol precursors, squalene and lanosterol, were totally ineffective in promoting NS-1 cell growth.

Difference in prognosis between T- and B-cell lymphomas: clinical study at Shikoku Cancer Center Hospital.

In order to determine whether the surface marker phenotypes of non-Hodgkin's lymphomas affect the prognosis, we have studied the differences in response rate and duration of survival between T- and B-cell lymphomas. Sixty-four patients who underwent first-line therapy, including combination chemotherapy and/or radiotherapy, from February 1979 to August 1985 were evaluated. With the aid of standard immunological methods and monoclonal antibodies related to T-cells and B-cells, 21 T-cell lymphomas and 21 B-cell lymphomas were identified.

Conservation of DNA sequences for plasmid-mediated citrate utilization within the enterobacteria.

Southern blot DNA-DNA hybridization experiments with a cloned Cit+ DNA fragment as a probe showed that the plasmid-mediated Cit+ determinants from four Cit plasmids (R726, pOH3001, pOH3035, and pOH30221) were all homologous. Sequences homologous to the plasmid-borne Cit+ gene were also found in total bacterial DNA isolated from Salmonella paratyphi B, Salmonella enteritidis, Salmonella typhimurium LT-2, Citrobacter freundii, ATCC 8090, Citrobacter amalonaticus ATCC 25405, Klebsiella pneumoniae I and IID 977, and Enterobacter aerogenes ATCC 13048. The DNA digest from C.

MDBK nuclear factor-binding site of various serotypes of adenovirus DNA.

A fraction with the ability to bind the terminal fragment of equine adenovirus (EAd) DNA was prepared from MDBK cell nuclei. The fraction (MDBK nuclear factor) bound to the terminal fragment of all human and animal adenovirus DNAs examined except avian adenovirus EDS-76. However, the terminal fragments of two animal adenoviruses, EAd and bovine adenovirus type 3 (BAd3), showed higher affinity for the nuclear factor than the others.

Purification of scrapie agent from infected animal brains and raising of antibodies to the purified fraction.

The fraction (P4) containing scrapie infectivity was obtained by treatment of scrapie-infected mouse brains with the detergent sarcosyl, differential centrifugation, and proteolytic enzyme digestion. Scrapie infectivity in the P4 fraction was purified 239-2,390 times with respect to protein. Similar fractions were also prepared from the brain of a sheep naturally infected with scrapie.

Nucleotide sequence of the gene determining plasmid-mediated citrate utilization.

The citrate utilization determinant from transposon Tn3411 has been cloned and sequenced, and its polypeptide products have been characterized in minicell experiments. The nucleotide sequence was determined for a 2,047-base-pair BglII restriction endonuclease fragment that includes the citrate determinant. This region contains an open reading frame that would encode a 431-amino-acid very hydrophobic polypeptide and which is preceded by a reasonable ribosomal binding site.

[An outbreak of staphylococcal dermatitis in laboratory mice].

An epizootic of dermatitis with erosion, ulcer and crust broke out in an experimental colony of JCL-ICR mouse over a period from December 1975 to June 1976. The disease was detected in 592 of a total of 1831 mice of 3-24 months old, especially in males of 7-24 months old (517/821). At the beginning of December 1975, only a few males of 12 months old were found to have the lesion on the back skin, and thereafter the dermatitis prevailed gradually among the mice.

Plasmid DNA relatedness among different serogroups of Yersinia pseudotuberculosis.

Thirteen different serogroup strains of Yersinia pseudotuberculosis and two strains of Yersinia enterocolitica O:3 were examined for the presence of plasmids and plasmid-mediated properties, calcium growth dependency, and autoagglutination. Two Y. enterocolitica strains and eight serogroup (IA, IIA, IIC, III, IVA, VB, VI, and VIII) strains, except for five serogroups (IB, IIB, IVB, VA, and VII), of Y.

[Biological properties, phage typing and antimicrobial susceptibility of Staphylococcus aureus isolated from dermatitis in laboratory mice].

The characteristics of 167 isolates of S. aureus from 106 mice suffering dermatitis were examined. All 167 isolates coagulated both rabbit and human plasmas and 161 of them also coagulated bovine plasma.

Characterization of scrapie agent isolated from sheep in Japan.

A pathogenic agent isolated in mice from the brain of a sheep affected by scrapie-like disease was characterized. The incubation period of the disease in the primary transmission from the sheep to mice was longer than in the secondary and the tertiary transmission in the same strain of mice. Progressive dilution of the inoculum caused prolongation of the incubation period.