One hundred and twenty-one children showing symptoms of acute upper respiratory disease were bled during a period of nine months, from winter to summer, for detection of complement-requiring neutralizing (CRN) antibody against herpes simplex virus. Six of the cases, all from children under the age of 13 years, were unequivocally related to herpetic infection as evidenced by the presence of anti-herpes CRN antibody. During the same period, 144 other children who were normal healthy or suffering from unrelated non-febrile diseases were tested as controls; anti-herpes CRN antibody was not detected in any of them. Further, the age distribution of individuals with antibodies was compared between patients in the acute upper respiratory disease group and the control group. This analysis showed that approximately 5 to 7% of the acute upper respiratory diseases in the young children in this study would be attributed to herpetic infection.It has been hitherto believed that, although a variety of disease manifestations may result from primary and repeated infections of herpes simplex virus in man, the lesions are limited mainly to the skin and the outer mucous membrane, with the exception of some unusual instances such as encephalitis or generalized infection in newborn infants [1,2]. A recent report of Scott and Tokumaru [3], however, revealed the occurrence of upper respiratory diseases due to herpes virus. Thus, herpes virus has been registered anew as a member of the family of respiratory viruses. As a result of this last report, the author attempted to estimate the importance of herpetic infections as a cause of acute upper respiratory disease (ARD) in children.In the present study, determination of herpes infection as a cause of ARD was done by detection of CRN antibody against herpes virus [5], since this method did not require paired serum samples for diagnosis and consequently enabled the handling of a large number of sera with ease. The results indicated that some of the ARD cases were related to herpetic infection as evidenced by this serological test. Further analysis was then made upon the age distribution of ARD patients and control children with antibodies, to test the possibility that some cases of herpetic infection were missed by this serological method of diag-

Plaque formation by the HF strain of herpes simplex virus in HeLa cells under agar overlay was possible when a line of this strain highly adapted to HeLa cells was used, whereas the homologous strain, passaged in eggs or lowly adapted to HeLa cells did not form any visible plaques using the same technique. Experiments were performed to investigate the influence of different diluents, cellular factors and conditions of virus adsorption. It was found that 0.1% yolk-saline could replace the ordinarily used maintenance medium as a diluent. Among three lines of S3 HeLa cells maintained in different laboratories, a subclone possessing a rapid cellular growth rate showed the highest sensitivity to the virus. Monolayers which scarcely covered the whole glass surface were most suitable for virus inoculation. Such monolayers could be stored at 20 C for 2 days before use without lowering the efficiency of plaque formation. The highest titer of virus was obtained when adsorption of inoculated virus was allowed to take place at 37 C for 2 hr. This titer was about equal to the plaque titer obtained in primary chick embryo cell cultures.

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Summary The three suspected cases of HSV encephalitis who still survive were reported. Case 1, a 8 months old girl, and case 2 of 8 years 11 months old girl were diagnosed with basis of clinical findings and clinical course, serological demonstration of greater than four‐fold rises in complement fixing antibody and non‐CRN antibody, significant rise in CRN antibody (CRN/non‐CRN) and tests which could rule out other common viral origin Case 3 was diagnosed by isolation of virus from C.S.F. obtained at intraventricular puncture and by a significant rise in antibody (CF antibody, non‐CRN and CRN antibody).