Negative motor networks: Electric cortical stimulation and diffusion tensor imaging.

INTRODUCTION: This study investigated the networks of Negative motor areas (NMAs) using electric cortical stimulation and diffusion tensor imaging (DTI). METHODS: Twelve patients with intractable focal epilepsy, in which NMAs were identified by electrical cortical stimulation, were enrolled in this study. Electric stimulation at 50Hz was applied to the electrodes during motor tasks to identify the NMAs. DTI was used to identify the subcortical fibers originating from the NMAs found by electrical stimulation. RESULTS: NMAs were found in lateral frontal areas (premotor area (PM) and precentral gyrus) in all 12 patients, in pre-supplementary motor areas (pre-SMAs) in four patients, and in posterior parietal cortices (PPCs) in four. DTI detected fibers connecting to the ipsilateral PMs, PPCs and temporal regions via U-fibers, superior longitudinal fasciculus (SLF), and arcuate fasciculus (AF) from the lateral frontal NMAs. Pre-SMA-NMAs had connections with ipsilateral PMs and contralateral pre-SMAs via the frontal aslant tract and transcallosal commissural fibers, and PPC-NMAs with ipsilateral PMs via SLF and AF. CONCLUSION: This study found the characteristic cortical network of each NMA, and especially revealed new insight of pre-SMA-NMA and PPC NMA. These NMAs might be associated with different mechanism of negative motor response.

Development of adult T-cell leukemia-like disease in African green monkey associated with clonal integration of simian T-cell leukemia virus type I.

Proviral integration of a simian retrovirus highly homologous to human T-cell leukemia virus type I was examined in cellular DNAs extracted from primary peripheral blood lymphocytes of 31 adult African green monkeys (Cercopithecus aethiops) that were seropositive for simian T-cell leukemia virus type I (STLV-I). Among these monkeys, one case with overt leukemia, showing pleomorphic leukemia cells similar to those in human adult T-cell leukemia (ATL), and five cases in a preleukemic state of ATL-like disease were found. Judging from the integration site of the provirus genome, primary lymphocytes of these leukemic or preleukemic cases contained monoclonally proliferated STLV-I-infected cells, whereas lymphocytes of other seropositive monkeys without hematological abnormalities were polyclonal, and those of seronegative monkeys did not contain the provirus. The restriction patterns with PstI ans SstI of most STLV-I proviruses were identical to those of the previous isolate from this species, but in three monkeys there was a deletion of one PstI site. From the correlation of the development of simian ATL-like disease with the monoclonal integration of the STLV-I provirus genome, it should be indicated that STLV-I has similar leukemogenicity to human T-cell leukemia virus type I, and so STLV-I infection in African green monkeys will be useful as an animal model of human ATL.

Characterization of simian viruses isolated from wild cynomolgus monkeys.

Twenty three strains of simian viruses isolated from nasopharyngeal swabs and respiratory tissues of cynomolgus monkeys showing mild symptoms of respiratory disorders were classified into three groups by their biological, serological and morphological characteristics. Thirteen strains classified as group 1 were judged to be enteroviruses. Four strains of group 2 and six strains of group 3 were identified as simian adenoviruses. Yields of the ten simian adenoviruses of groups 2 and 3 were compared in three continuous cell lines after several serial passages.

[Effectiveness of hypoallergenic rice-by cast analysis].

Rice is the main food taken every day in Japan. We introduced hypoallergenic rice (95% of albumin and globulin were taken off by hyperpressure method) for 4 weeks to 7 atopic dermatitis patients (m 3, f 4), 11 mo-4 y 8 m (mean 3 y 2 m) who showed the RAST score of rice 1 or more. Stimulation Index by CAST (cellular antigen stimulation test) were compared before and after taking the rice (50-100 g). By albumin (0.1-100 micrograms/ml), maximum decreases of S.I. were 22.4-96.5% (mean 70.9) and by globulin maximum decreases of S.I. were (0.1-100 micrograms/ml) 33.3-97.0% (mean 72.9). In all the patients the skin conditions were improved.

Enterovirus type 70-neutralizing IgM in animal sera.

The neutralizing activity against human enterovirus type 70 was found in serum samples from normal cattle, horses, sheep, goats, swine and chickens raised in Japan. The frequency was variable depending upon animal species and the year of bleeding. The neutralizing activity in bovine sera was shown to reside in IgM by sucrose gradient centrifugation and immune gel electrophoresis. These findings suggested that the neutralizing substance in domestic animal sera is the antibody of IgM class elicited by unidentified viruses antigenically related to human enterovirus type 70.

Serologic characterization and sero-epidemiologic studies on acute hemorrhagic conjunctivitis (AHC) virus.

Serologic and sero-epidemiologic characteristics of AHC virus infection were studied by neutralization test (NT). Four-fold or greater virus neutralizing (VN) antibody response was demonstrated to the Japanese isolate of AHC virus (the J 670/71 strain) in 77.3% and 66.7% of paired sera from clinical AHC patients in Japan (1971-1973) and Tunisia (1973). The four patients from Indonesia studied in 1972 showed similar antibody response. Cross-neutralization tests of AHC virus isolated in Japan (1971), Taiwan (1971), Hong Kong (1971), Thailand (1972), Indonesia (1972), Singapore (1972), Morocco (1971) and England (1971) with three kinds of antisera prepared against Japanese, Hong Kong and Moroccan AHC virus isolates indicated their antigenic identity. However, isolates from Sinapore in 1970 (Singapore 70 virus) were not neutralized with the AHC virus antisera mentioned above: Singapore 70 virus constitutes another antigenic type, to which, however, no VN antibody rise was found in paired patients' sera from Japan, Tunisia and Indonesia. Thus, no serologic evidence supporting an etiologic role of this virus group in the development of AHC was found. Although cross-tests using monospecific antisera suggested some cross-relation between AHC and both echovirus type 4 (E4) and coxsackie A (CA), type 19, no serologic relationship between AHC and these viruses was found. Sera from healthy individuals collected before and after AHC outbreaks were tested for VN antibody against AHC virus in Japan and two epidemic foci, Ghana and Indonesia. Before the epidemic, 80 to 90% of the people lacked antibody in the three countries, but 39.7% and 45.2% of inhabitants posessed VN antibody of 1:8 or over in Ghana and Indonesia after the outbreak. In Japan, however, only a slight increase was found in VN antibody prevalence afterwards. Serologic study showed that 41.5% of horse sera were VN positive at dilutions of 1:8 or more; many cattle sera also had a low VN titer but few cynomologus monkey sera had VN activity.

Growth characteristics of acute hemorrhagic conjunctivitis (AHC) virus in monkey kidney cells. II. Temperature sensitivity of the isolates obtained at various epidemic areas.

Acute hemorrhagic conjunctivitis (AHC) virus strains isolated in eight different areas during epidemics of AHC were tested for the reproductive capacity at 33, 37 and 39 degrees. All of the 25 strains tested grew better at 33 degrees but restrictively at 39 degrees. The degree of temperature sensitivity varied slightly from one strain to the other, but generally exceeded that of attenuated poliovirus type 1, strain LSc2ab. Temperature-resistant clones were selected by repeated passages of originally temperature-sensitive prototype strains at supraoptimal temperature. The importance of using a low temperature (32-34 degrees) for isolation of virus from external tissues of the body and for subsequent passages has been emphasized. It was suggested that the relatively low temperature of the conjunctiva has played a role in perpetuating temperature sensitivity of this virus.

Absence of antibodies against SV5, measles and herpes simplex in indoor colony-bred cynomolgus monkeys.

One hundred and eighty-one cynomolgus monkeys born and raised at Tsukuba Primate Center for Medical Science (TPC) were serologically examined for the presence of antibodies against simian virus 5, measles virus and herpes simplex virus. All of them were virtually free from infections with these viruses, whereas wild-originated breeders kept at TPC were highly positive for these antibodies. These results can be regarded as demonstrating that an indoor breeding system such as TPC's is useful and valuable for producing nonhuman primates of good quality being completely free from those viral infections.

Serological survey for SV5, measles and herpes simplex infections in newly-imported cynomolgus monkeys.

On 592 cynomolgus monkeys of newly-imported 14 different groups, sero-epidemiological follow-up surveys of natural infections with SV5, measles (MV) and herpes simplex (HSV) were conducted both at the time of arrival and at the end of the 9-week quarantine period at Tsukuba Primate Center for Medical Science (TPC). At the time of arrival, the positive rate of antibodies against these viruses greatly varied with different countries of origin. As regards SV5, the positive rate ranged widely from 0 to 94.9% among different import-groups, being markedly low in the Philippine monkey groups. The positive rate of MV antibody was generally very high, varying from 11.1 to 78.2% in almost all import-groups except two groups from Indonesia in which no positive case was detected. Concerning HSV, every import-group had a very high positive rate though the average antibody titer was not so high. Generally, the positive rates of SV5 and MV markedly increased by the end of quarantine period. Especially, MV positive rate reached 100% in almost all of the import-groups. However, two Indonesian groups showed no increase of SV5 and MV positive rates during the quarantine period. No increase of SV5 positive rate was detected either with any of three Philippine groups. As for HSV, the positive rate remained unchanged or rather decreased during the quarantine period in almost all groups except two Indonesian groups having shown a significant increase.

Hematologic abnormalities similar to the preleukemic state of adult T-cell leukemia in African green monkeys naturally infected with simian T-cell leukemia virus.

Studies were made of the hematologic features of 23 adult African green monkeys (Cercopithecus aethiops) naturally infected with simian T-cell leukemia virus (STLV), which is closely related to human T-cell leukemia virus type I. None of the STLV-infected monkeys showed any clinical signs, but their absolute lymphocyte count and percentage of atypical lymphocytes were significantly higher than those of uninfected monkeys. Three STLV-infected monkeys had especially high percentages of atypical lymphocytes. Most of the atypical lymphocytes were small- or medium-sized lymphocytes with a lobulate or convoluted nucleus, and were very similar to the leukemic cells in human cases of adult T-cell leukemia (ATL), while other atypical cells were large lymphoblastoid cells. Both types of atypical cells were positive for Leu2a and Tac antigens, and expressed STLV antigen after short-term culture. These findings suggested the presence of a preleukemic state in these STLV-infected monkeys similar to that seen in human ATL, and indicated the value of this natural infection system as an animal model of ATL in humans.

Herpes simplex virus complement fixing antibody and herpes B virus serum neutralizing antibody in sera of wild and laboratory-bred cynomolgus monkeys.

The result of the complement fixation (CF) test for the antibody to herpes simplex virus (HSV) in sera of the cynomolgus monkeys was compared with that of the neutralization test (NS) for the antibody to herpes B virus (HBV) in the same sera. Fifty-seven (74%) of 77 wild-originated monkeys were positive for HSV-CF, while 65 (84%) of the 77 animals were positive for HBV-SN. All of the 57 CF positive cases were also positive for HBV-SN. On the other hand, 30 (75%) of 40 laboratory-bred monkeys had neither HSV-CF antibody nor HBV-SN antibody. Remaining 10 of the 40 laboratory-bred animals were positive for HSV-CF. However, no HBV-SN antibody was detected in nine of the 10 HSV-CF positive animals. These results suggest that the HSV-CF test may be as satisfactory as the HSV-SN test as a practical measure for rough screening of HBV infection in the cynomolgus monkey in laboratories having no containment unit for handling HBV.

Hemoagglutination and hemagglutination inhibition tests with enterovirus type 70.

Human type "O," guinea pig, and chicken erythrocytes were agglutinated by enterovirus type 70 at 4 degrees C or room temperature. A hemagglutination inhibition test, using human "O" erythrocytes, is described for the serological diagnosis of acute hemorrhagic conjunctivitis caused by enterovirus type 70.

Spontaneous malignant lymphoma in an African green monkey naturally infected with simian T-lymphotropic virus (STLV).

An African green monkey naturally infected with simian T-lymphotropic virus (STLV) developed spontaneous malignant lymphoma of diffuse pleomorphic type. The clinical, hematological and histopathological characteristics were very similar to those of human adult T-cell leukemia.

Virological and serological studies of neurological complications of acute hemorrhagic conjunctivitis in Thailand.

An extensive outbreak of acute hemorrhagic conjunctivitis (AHC) occurred from September to December 1974 in Thailand. At least 29 patients with polio-like motor paralysis that complicated AHC were hospitalized in Bangkok. Paired or triplicate samples of serum from 16 patients were tested for neutralizing antibody to enterovirus type 70 (EV70). A significant rise in titer of antibody was found for two patients, and the other 14 had neutralizing antibody titers ranging from 1:8 to 1:512 without an increasf larger than or equal to 1:16, a level which is considered to be diagnostically significant. Neutralizing antibody to EV70 was detected in 19S fractions of nine sera examined, but neutralizing antibody to three types of poliovirus was confined to 7S fractions. EV70 was isolated from one of seven stool specimens collected on day 37 after the onset of AHC and none of 10 samples of cerebrospinal fluid. These results and additional clinical and epidemiologic findings gave further support to the hypothesis that EV70 infection can cause polio-like motor paralysis as a complication of AHC.

Adult T-cell leukemia-like disease in monkey naturally infected with simian retrovirus related to human T-cell leukemia virus type I.

Spontaneous T-cell leukemia similar to human adult T-cell leukemia (ATL) was found in an African green monkey naturally infected with simian retrovirus closely related to human T-cell leukemia virus type I (HTLV-I). Monoclonal integration of the simian retrovirus was detected in the primary leukemic cells, suggesting an association of the retrovirus with ATL-like leukemia in the monkey.

Protection of cynomolgus monkeys against infection by human T-cell leukemia virus type-I by immunization with viral env gene products produced in Escherichia coli.

Protection against human T-cell leukemia virus type-I (HTLV-I) infection in cynomolgus monkeys, achieved by immunizing the animals with env gene products of HTLV-I produced in Escherichia coli, was evaluated. Four monkeys that had been immunized with the env product produced antibody against HTLV-I gp68 and gp46, and their sera were found to cause strong inhibition of syncytium formation of a cat fibroblast cell line induced by HTLV-I. Immunized and non-immunized monkeys were challenged with live MT-2 cells, a high HTLV-I-producer cell line. After challenge, all the control non-immunized monkeys were infected with HTLV-I, as judged by the frequent detection of HTLV-I-antigens in cultures of their peripheral blood mononuclear cells (PBMC), whereas no antigens were recovered from PBMC of immunized monkeys. These results indicate that humoral immunity against HTLV-I-envelope protein elicited by immunization with the polypeptides synthesized in bacteria protected the monkeys against primary infection with HTLV-I.

Seroepidemiologic studies of acute hemorrhagic conjunctivitis virus (enterovirus type 70) in West Africa. II. Studies with human sera collected in West African countries other than Ghana.

Human sera were collected in Senegal, Nigeria, Ivory Coast, Dahomey, Liberia, Gabon and Togo during the pre-epidemic period of acute hemorrhagic conjunctivitis (AHC) from 1965 to 1969, and tested for virus neutralizing (VN) antibody to enterovirus type 70 (EV70). Of these, 1109 (91%) were antibody negative (less than equal to 1:4), 116 (9%) neutralized at a dilution of 1:8 or over, and 45 (4%) at dilutions of at least 1:16. The distribution pattern is not significantly different from that of sera collected from Kenya in 1967 or from army recruits in the United States, Argentina, Brazil and Colombia in the 1960s. Sera collected during the post-epidemic period (1970 to 1977) in Senegal, Sierra Leone, Mali, Upper Volta, Chad, Niger and Gabon were also examined; 1573 (68%) were VN antibody negative (less than or equal to 1:4), while 733 (32%) and 433 (19%) had titers of 1:8 or greater and 1:16 or over, respectively. There is a significant difference in distribution between pre- and post-epidemic antibody titers (p less than 0.001), although the incidence of AHC was lower in these countries than in Ghana and Southeast Asia. The prevalence of VN antibodies tends to be lower in the dry, hot inland areas and thus humid coastal monsoonal climates and dense populations seem to favor the spread of AHC.

Seroepidemiologic studies of acute hemorrhagic conjunctivitis virus (enterovirus type 70) in West Africa. III. Studies with animal sera from Ghana and Senegal.

One hundred and thirty-nine bovine sera collected in Senegal in 1968 and 1969, before the human pandemic of acute hemorrhagic conjunctivitis (AHC), and 145 sera collected in 1977, seven years after the introduction of AHC, were tested for virus neutralizing (VN) titers against enterovirus 70 (EV70) in neutralization tests. Positive rates of VN titers (1:16) were fairly constant (about 40%) in 1968 and 1969 but the proportion of positives from the 1977 collection was significantly higher (61%). The proportion of Ghanaian bovine sera positive in 1977 (39%) was comparable with those collected earlier in Senegal but the proportion was lower in sera of calves (7%). Similar studies were performed on sheep sera collected in 1966, 1967, 1968, and 1969 in Senegal. The proportions positive and the geometric mean titers (GMTs) peaked in alternate years: the proportions were 43% and 54% in 1966 and 1968 but dropped to 12% and 5% in 1967 and 1969, respectively. The prevalence rate for Ghanaian sheep sera in 1977 was comparable to the earlier figures in Senegal. The prevalence of VN (83%) and the GMT (1:27.28) in swine sera in Ghana in 1977 were much higher than those seen in chickens and dogs. However, 87 sera of wild monkeys caught in Senegal after 1970 were negative in neutralization tests. The VN substance detected in these domestic animals was found to be 2-mercaptoethanol sensitive; hence it is considered to belong to IgM. The results seem to favor the hypothesis that enterovirus type 70 (EV70) evolved from an animal enterovirus which shares a common antigen with EV70.

Comparison of gelatin particle agglutination and hemagglutination inhibition tests for measles seroepidemiology studies.

The prevalence of measles antibody in Japan was surveyed with a newly developed gelatin particle agglutination (PA) test, and the results compared with those of the hemagglutination inhibition (HI) test. The two age-distribution curves of the PA antibody-positive rates at > or = 1:8 and > or = 1:32 were almost the same in all the age groups, except the less-than-1-year-old group for which the rate at > or = 1:8 was higher than that at > or = 1:32 (p < 0.05, chi 2 test). In the vaccinated children, all groups older-than-1-year of age had antibody-positive levels of 96% or more. In contrast, in the unvaccinated children, there was a sharp increase in antibody-positive rates between the 1- and 4-year-old groups, indicative that about 80% of the children were infected by wild measles virus at these ages. A significant number of PA antibody-positive specimens were antibody-negative (< 1:8) by HI. The percentage of specimens in this category, PA (+) but HI (-), was greatest in infants less than one year old, and least in young children, but it increased with age to 97% of the HI (-) specimens from adults of more than 20 years of age. The PA test therefore detected some measles antibodies that HI could not. This test is simple and useful for making serosurveys in both developed and developing countries.