Biomedical subjects
I Arita
Publications and source records attributed to I Arita.
X-irradiation induces up-regulation of ATM gene expression in wild-type lymphoblastoid cell lines, but not in their heterozygous or homozygous ataxia-telangiectasia counterparts.
Ataxia-telangiectasia (AT) is an autosomal recessive disease. The relevant gene has been cloned and designated ATM. We studied the expression of both ATM mRNA and the ATM protein in unirradiated and X-irradiated EBV (Epstein-Barr virus)-transformed lymphoblastoid cell lines (LCLs) derived from donors who were normal (ATM + / + ), AT heterozygotes (ATM + / - ), or AT homozygotes (ATM - / - ), respectively. In ATM + / + LCLs, the levels of ATM mRNA were found to have increased by approximately 1.5-fold within 1 h of exposure to 10 Gy of X-rays, while the ATM protein levels had increased by 1.5- to 2.0-fold within 2 to 3 h of irradiation. The wild-type mRNA and protein levels both returned to their basal values fairly quickly after this time. The results obtained with the ATM + / - LCLs were quite different, however: neither the mRNA nor protein levels were found to have increased as a consequence of X-irradiation in any ATM + / - LCL. Twelve of the mutations in the ATM - / - LCLs we used were truncating mutations, and we suspected that the corresponding truncated ATM proteins would be too labile to be detected by western blot analysis. However, five of the ATM - / - LCLs produced mutant ATM proteins that were identical in molecular weight to the wild-type ATM protein. When cells from three of these five clones were exposed to X-rays, transcription of the mutant ATM genes appeared to reduce somewhat, as were the levels of protein being produced. These results suggest that the normal ATM gene responds to ionizing radiation by up-regulating its activity, whereas none of the mutant ATM genes we studied were able to respond in this way.
A scenario for polio eradication.
Polio eradication has entered the phase operating in the Asian sub-continent and in sub-Saharan Africa, where polio is still endemic in 30 countries. These countries are apparently most resistant to the eradication effort. The WHO polio panel held in May, in Geneva, discussed a scenario to stop transmission by 2002 and thereafter three years for certification. This paper discusses the scenario based on experience with smallpox eradication from the viewpoint of impact of population density on immunisation. The programme in the Indian sub-continent, which has the world's highest population density, may require a more rigorous surveillance-containment type strategy than that in sub-Saharan Africa which, with its lower density of population in many countries, may be relatively susceptible to the eradication effort. However, countries with higher population density and/or with political trouble may pose substantial difficulties. Also briefly discussed are measures to be taken after interruption of polio transmission world-wide. These still require intensive studies. The goal is foreseeable and with intensified mobilisation of global resources.
Does polio eradication succeed meeting the target year of 2000?
The global polio eradication program is under way with the target year of 2000. We reviewed the program progress, in perspective, from the experience of the global smallpox eradication, which is at present a sole disease ever eradicated by orchestrated global efforts. We concluded that despite substantial efforts being made by the World Health Organization and member states with the current progress, it would require additional some three years, namely 2002. This would safe-guard the success. As of January 2000, there are still at least 22 endemic states in Indian Subcontinent and sub-Saharan Africa. Experiences in smallpox eradication, although the disease is different, indicated that to stop transmission in such a large number of endemic states took more than three years, assuming that further support would come toward the final goal.
Standardization of smallpox vaccines and the eradication programme--a WHO perspective.
Smallpox vaccine was born in 1796 and the need for it ended in 1980 with the eradication of smallpox. What was remarkable was that in its 200 years history, when the global smallpox eradication programme really needed the vaccine in 1967, it was found that, on the whole, quality was unsatisfactory and supply was short. Unconventional counter measures were taken by WHO, including an international independent testing system, provision of reference preparations in large quantity etc. They effectively solved the problems meeting the target eradication schedule of 10 years. The lessons learned would be of some value to the future eradication or effective control programme of vaccine preventable diseases.
The global capacity for manufacturing vaccines. Prospects for competition and collaboration among producers in the next decade.
Can the world respond to the demands of the Children's Vaccine Initiative (CVI) to produce large quantities of affordable vaccines that have never been manufactured previously? Vaccines for the world's birth cohort of 150 million will continue to be produced in the countries that use the greatest part of the global vaccine supply. Thus, the CVI will rely on increased self sufficiency in vaccine production in the developing world and "shared development" of new and improved vaccines. The CVI's goal is to direct product development to meet the needs of immunization programs, but it must not neglect production. Thus, from the start, investment at the front end of the development and production sequence requires attention to the ultimate production capacity.
Transition of phenotypic dimorphism with regard to spontaneous sister chromatid exchange in Epstein-Barr virus-transformed Bloom's syndrome lymphoblastoid cell lines.
We recently established four lymphoblastoid cell lines (LCLs) by infecting the peripheral blood of four Japanese patients suffering from Bloom's syndrome (BS) with Epstein-Barr virus (EBV). During the course of propagating these cell lines, two of them exhibited dimorphism regarding spontaneous sister chromatid exchange (SCE), i.e., a mixed population consisted of cells with extremely high SCE levels characteristic of BS and cells with low SCE levels indistinguishable from that of normal control cells. On the other hand, the other two cell lines maintained a monomorphic population with high SCE levels at least until 30 weeks after EBV infection. The proportion of the cells with high SCE levels in the cell lines with dual phenotype declined as the population doubling numbers (PDN) increased with time and they became ultimately undetectable. The proportion of cells with low SCE levels at the time of EBV infection was estimated in one of these LCLs as 0.075% by extrapolating the linear regression of the logit for the proportion plotted against PDN. In view of the well-known stability of the monomorphic phenotype in representative BS LCLs during extended cultivation, together with the present observations on the dual phenotype, we conclude that the frequent establishment of BS LCLs exclusively with low spontaneous SCE levels is attributable to the various proportions of low-SCE cells existing in vivo in the B-lymphocytes pool of BS individuals and to the selective pressure against the high-SCE cells in in vitro cultures.
Antibodies to hepatitis C virus in Japanese blood donors and in hospital personnel.
Explore the source record for details and available documents.
Evidence for spontaneous conversion of Mex- to Mex+ in human lymphoblastoid cells.
A series of human lymphoblastoid cell lines (LCLs) called Mex- were defined by Sklar and Strauss on the basis of their inability to remove O6-methylguanine from DNA. Instability of Mex- has previously been shown as a population phenotype of LCLs. We examined whether Mex- as a cellular phenotype is spontaneously convertible or not. At the population doubling number (PDN) 23 after recloning, two out of 15 independent subcultures derived from a Mex- LCL, AT1-1, were found to contain a small fraction of Mex+ cells after treatment with 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU). Three Mex+ subclones were identified without exposure to ACNU among 486 subclones from replica plating of an expanded Mex- clone (PDN30). The rate of spontaneous conversion was estimated to be in the range of 10(-8)-10(-7) per cell per generation by the fluctuation analyses on two Mex- subclones. These results strongly support the hypothesis that Mex- as a cellular phenotype is spontaneously convertible to Mex+.
Tumorigenic conversion of xeroderma pigmentosum lymphoblastoid cells without karyotypic alteration.
In order to examine the process of malignant transformation of human somatic cells, we studied the tumorigenic conversion of an Epstein-Barr-virus-immortalized lymphoblastoid cell line (LCL) derived from a patient with xeroderma pigmentosum (XP) complementation group A. Repeated irradiation of the XP cells, XP7NI, with UV-light and subsequent treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in the acquisition of tumorigenicity in athymic nude mice. The tumorigenicity of XP7NI cells was also induced by TPA treatment alone. The tumors formed in athymic mice were of B-cell lymphoma with characteristic histology, cell surface immunoglobulins and an antigen as detected by a B-cell-specific monoclonal antibody (MAb), CD20. The surface immunoglobulins and the HLA type of these tumor cells were identical with those of the parental cells. These malignantly transformed cells retained the same UV sensitivity, serum requirement, colony-forming ability in soft agar, and normal human karyotype as the parental cells. Unlike other tumorigenic lymphoblastoid cell lines, this XP lymphoblastoid cell line provides a unique case in that process(es) leading to tumorigenicity may be induced by UV and TPA without apparent karyotypic changes.
Predominance of Mex+ cells in newly-established human lymphoblastoid cell lines.
Previous studies have demonstrated that approximately one-third of human lymphoblastoid cell lines (LCLs) are deficient in removing O6-methylguanine residues because of the lack of O6-alkylguanine-DNA alkyltransferase (O6-AGT) activity. Such LCLs have been designated Mex-, while the proficient LCLs are Mex+. Our determinations of O6-AGT activity as a function of cellular protein concentration on 37 previously-established LCLs disclosed that the expression of the enzyme was high in 14 (Mex+) and barely detectable in 16 (Mex-). The other seven LCLs showed intermediate activity of the enzyme. By contrast, all of the 28 LCLs that we newly established contained high enzyme activity, implying that they consisted of mainly Mex+ cells. Since the conventional O6-AGT assay on Mex+ cell populations was not capable of detecting the co-existence of Mex- cells as a minor component, we attempted to determine the proportion of Mex- phenotype in newly-immortalized lymphoblastoid cell clones which had been established directly on semisolid agar. All of the 15 independent clones derived from a single blood sample also showed high O6-AGT activity, rendering it unlikely that Epstein-Barr virus transformation per se was responsible for the generation of Mex- LCLs. These results collectively indicate that Mex+ cells predominate in LCLs shortly after establishment and also suggest that the possible growth advantage for Mex- cells should play an important role in the subsequent development of Mex- LCLs during the long-term culture in vitro.
Pseudohypoparathyroidism showing positive phosphaturic and negative cyclic AMP excretion response to parathyroid hormone.
We report a patient with pseudohypoparathyroidism (PHP) in whom parathyroid hormone (PTH) infusion failed to produce an increase in urinary adenosine 3', 5' monophosphate (cAMP) excretion in spite of the positive urinary phosphate excretion. The dbcAMP infusion test showed almost the same increase in phosphate as in the E-H test, although high urinary cAMP excretion was detected. Furthermore, a PTH infusion test in combination with calcium antagonist (diltiazem) administration markedly increased phosphate excretion, whereas the response of urinary cAMP excretion also remained negative. After treatment with 1 alpha(OH)D3, phosphaturic response increased by at least 14.3 mg/2 h compared with that in the pretreatment period. Therefore, intra and extra cellular calcium seem to affect the phosphaturic response induced by PTH.
Instability of Mex- phenotype in human lymphoblastoid cell lines.
Three lymphoblastoid cell lines (LCLs) had extremely low activities of O6-alkylguanine-DNA alkyltransferase (O6-AGT), and were classified as Mex-. They were highly sensitive to cell killing by 1-(4-amino-2-methyl-5-pyrimidinyl)-methyl-3-(2-chloroethyl)-3-nitrosoure a hydrochloride (ACNU), whereas NMO2, a Mex+ LCL with a high O6-AGT activity, was resistant to the agent. Small fractions of these Mex- LCLs survived the treatment with 10 micrograms/ml of ACNU for 24 h, and the surviving cells were found to be resistant to subsequent treatments with the agent. In addition, they contained O6-AGT activities comparable to that of NMO2 and were therefore regarded as Mex+. These results suggest that the Mex- phenotype in LCLs is unstable.
Serological survey for human monkeypox infections in a selected population in Zaire.
About 3460 persons living in Kole zone of East Kasai, in Zaire, were examined and their sera screened initially by a haemagglutination-inhibition test. Of these, 667 (19%) were positive. Radioimmunoassay adsorption tests for the presence of monkeypox- or vaccinia-specific antibodies gave unequivocal results in 300 of these sera; the remaining 47 were nonspecific. Monkeypox-specific antibodies were found in sera of 27 individuals, of all ages and both sexes, giving an overall prevalence rate of monkeypox virus-specific antibodies of 0.8%. The prevalence rate was four times higher in the 5 to 9 year age group (1.3%) than in children aged 0 to 4 years (0.3%), and was highest (2.4%) in the 15 to 19 year age group. There was no significant difference in the prevalence rates between the sexes. As might be expected, there are substantially higher prevalence rates in persons living in forest galleries than in those in savannah, and among those living in areas where human monkeypox cases had occurred in the past compared with those living in other localities. Nineteen children whose sera showed specific monkeypox antibodies were re-examined. Twelve showed facial and body skin changes suggesting the presence of vesiculo-pustular disease in the past; four of these had been known registered monkeypox cases. Seven children had neither signs nor history of past vesiculo-pustular disease, suggesting that they had suffered from subclinical infection with monkeypox virus.
Impact of population density on immunization programmes.
The eradication of smallpox was achieved by surveillance and containment vaccination after the failure of mass immunization campaigns. The reasons for this failure are considered in this paper. Comparison of population densities in the Indian subcontinent and Africa show that in highly populated areas even an 80% vaccine coverage will still leave a density of susceptibles high enough to maintain the disease, a finding with important implications for other vaccine campaigns.
Four generations of probable person-to-person transmission of human monkeypox.
This paper examines an outbreak of five cases of human monkeypox which occurred in children belonging to two families living in the West Kasai region of Zaire during May-July 1983. Epidemiologic investigations suggest that the first case was infected from an animal source, possibly a monkey, and that each of the other four cases was infected from a previous human case. Three of these cases of presumed person-to-person transmission occurred in close household contacts. The other case infection occurred either by casual contact within the hospital compound, or possibly because of infection due to use of the same syringe for injections. Human monkeypox is the most important orthopoxvirus infection in the post-smallpox eradication period. The disease is a zoonosis and person-to-person transmission is rather difficult. Thus, this episode is a rare event and special analysis of the circumstances is discussed. However, it supports the necessity to carry out surveillance and research on this disease as recently reported by Arita et al.
Human monkeypox: a newly emerged orthopoxvirus zoonosis in the tropical rain forests of Africa.
During the course of the recently concluded smallpox eradication program, a new human orthopoxvirus infection was discovered which is caused by monkeypox virus. The disease occurs sporadically in remote villages within tropical rain forests of West and Central Africa. The disease is rare; only 155 cases having been reported from 1970 to 1983. The symptoms and signs of human monkeypox resemble those of smallpox, differing significantly only in the occurrence of lymphadenopathy with human monkeypox disease. Of 155 cases, some 80% are believed to have resulted from infection from an as yet unknown animal reservoir; the rest occurred among unvaccinated close contacts among whom a secondary attack rate of 15% was observed. Although person-to-person spread appears to have occurred in some instances, few cases were observed in the third or fourth generation of transmission and none thereafter. Since 1982, the incidence of human monkeypox infections in Zaire has increased concomitant with an intensified surveillance program. Additional reasons which might explain the increased incidence are discussed. Further surveillance and research of this primarily zoonotic infection are warranted and are in progress.