Kazuko Araki

Infantile convulsions with mild gastroenteritis

The development of sensitive new molecular genetic techniques has led to the detection of rotavirus in cerebrospinal fluid, stools and throat swabs from patients with gastroenteritis with accompanying clinical symptoms similar to infantile benign convulsions. Small round structured virus (SRSV) has also been found in stools of patients with similar clinical symptoms by a new procedure. However, the mechanism by which these viral infections induce benign convulsions remains to be elucidated. The present paper reviews recent virological and clinical studies of seizures probably caused by gastroenteritis viruses including rotavirus, SRSV and other viruses.

Epidemiology of enteric adenoviruses 40 and 41 in acute gastroenteritis in infants and young children in the tokyo area

82/2,223 stool specimens, collected 1982-1988, from children with enteritis (3.7%) were found to contain adenoviruses; 17 adenovirus-positive samples were provided from other institutes. 89 adenoviruses were isolated in Graham 293 cells from these 99 specimens and were typed by DNA restriction enzyme analysis with Sma I. 37 strains were typed as adenovirus 40 (AD40), and 37 strains as adenovirus 41 (Ad41). Although most strains had the same DNA profiles, a few strains had 3 kinds of different electropherotypes generated by Sma I. Five strains were identified as adenovirus 31. The remaining 10 strains were adenovirus 1 (2 strains), adenovirus 2 (3 strains), adenovirus 3 (1 strain), adenovirus 5 (1 strain), and a non-classified adenovirus (3 strains). Ad40 and Ad41 infections were found throughout the year, but peaked between September and November. 80% of the children with adenovirus infections were less than or equal to 2 years of age. The highest incidence of diarrhea caused by Ad40 or Ad41 was in 6-11 months old children. 1982-1984, the rate of Ad40 infection was 91.7%, while the rate of Ad41 infection was only 8.3%. The prevalence of Ad40 infection gradually diminished from 1985. During 1987 and 1988 the reverse ratios, 20.6% and 79.4%, respectively, of Ad40 and Ad41 infections were observed. Thereafter, Ad41 infection became predominant.

Epidemiology of rotavirus infection in Tokyo during two winter seasons, as revealed by analyses of recovered viral RNA

Sir, - Rotaviruses are important etiologic agents of acute gastroenteritis in young children all over the world [9]. Gel electrophoresis can be used to detect and genotype rotavirus strains [2, 3, 4, 8]. We have investigated the relationships between rotavirus RNA electropherotypes and the epidemiology of acute gastroenteritis in Tokyo. Three hundred and seventy-four diarrheal stool specimens were collected from in- and out-patients (less than 4 years old, mainly between 6 months and 2 years old) at Teikyo University Hospital in Tokyo during the winter periods of 1981-1982 and 1982-1983. Outbreaks of rotavirus infection developed in two nursery rooms during this time. Twentyeight and 11 stool specimens were obtained from children in each nursery room, respectively. The extraction of virus from feces and

Coproantibody response to rotavirus in an outbreak in a day-care nursery

13 months and 10years 8 months. Patient no. 1 was first observed at a very advanced stage of the disease and died 19 months after the diagnosis of lung metastases. Patient no. 2 is still alive 10 years and 8 months after the diagnosis. Patients 3 and 4 are still alive after 6 years 11 months and patient no. 5 is well 6 years after the diagnosis. All the patients treated in the second period are still alive. Patients 6 and 8 are still in complete remission 35 and 13 months, respectively, after diagnosis. In patient no. 7 (alveolar RMS) a local relapse was noted 2 months after the completion of initial therapy. The subject was again treated with 5000 rads to the orbit, plus chemotherapy consisting of cisplatinum and tenixposide. After 7 months of treatment there were side-effects of renal toxicity with hyperuricaemia, increase in BUN and hyaline casts in the urine. Treatment was stopped and the patient is still in complete remission. Our results do not demonstrate any significant difference in life expectancy between patients treated with radical surgery and those treated conservatively, but the advantage of the latter treatment is so impressive cosmetically that this approach must be seriously taken into consideration. In our patients, the follow-up period is obviously too short to allow evaluation of long-term evolution and to draw definitive conclusions about the most suitable chemotherapy protocol. However, taking the quality of life, vision and psychological situation into consideration, we believe that chemotherapy and radiotherapy can offer a very valuable alternative in the therapy of RMS. Further trials involving this approach must be carefully considered.

A study of adenovirus gastroenteritis in the Tokyo area

Probable serotypes of faecal adenoviruses 40, 41, 31, 2 and 1 were found in the Tokyo area by rapid DNA restriction endonuclease analyses directly of stool specimens or after primary culture in Graham 293 cells. Some variations were found in both adenoviruses 40 and 41 by this method.

Comparison of five methods for detecting human rotavirus in stool specimens

Sir, Various methods have been developed for the detection of rotaviruses in stool specimens. For the rapid diagnosis of rotavirus infections, a simple latex agglutination (LA) test has recently become commercially available (Rotalex, Orion). Its sensitivity is comparable with that of polyacrylamide gel electrophoresis (PAGE) [3], and is four times greater than that of EM [11. We identified rotaviruses by electron microscopy (EM) after negative staining, by solid-phase immune microscopy (SPIEM) with protein-A [4], PAGE [5], LA, and reversed passive haemagglutination (RPHA) [2]. For the LA, antihuman rotavirus (strain Wa) rabbit serum was coated on latex beads. Stool samples were diluted with a solution of 0.2M Tris-HC1, 0.9% NaC1, and 0.1% bovine serum albumin. A commercially available RPHA kit was used (ROTA RPHA, Denka Seiken, Japan) with antihuman rotavirus (Wa strain) rabbit serum as antibody. Of 51 samples, 28 were positive and 18 were negative by all five techniques (Table 1). One sample was positive by four methods and negative by one, while one sample was negative by four methods and positive by one. The sensitivities of the tests were as follows: 30/51 (59%) for EM and SPIEM, 32/51 (63%) for PAGE, 31/51 (61%) for LA, and 29/51 (57%) for RPHA. EM and SPIEM were less sensitive than PAGE and LA. Although PAGE is the most sensitive technique, it takes 2-3 days before RNA genomes are visualized. All LA-positive specimens were positive in EM, SPIEM and/or PAGE. Though RPHA is the least sensitive test it is valuable for screening a large number of samples. LA is suitable for rapid bedside detection of rotavirus in stool specimens. This work was supported in part by the Ministry of Education Grant-in-Aid for Scientific Research 59440048, Japan, and by a grant from the Morinaga Hoshikai of Japan.

Evaluation of four tests for detecting human rotavirus in feces

Sir, Infantile diarrhea during the winter is mostly due to viruses and the majority of pathogenic viruses prove to be rotavirus. Various methods for diagnosing infections by rotavirus have been described. The migration patterns of RNA segments ascertained by polyacrylamide gel electrophoresis (PAGE) are sensitive for the detection of rotavirus and effective for the analysis of the molecular epidemiology of rotavirus infections [9]. A simple and specific technique for rapidly detecting rotaviruses in feces is still needed. Enzyme-linked immunosorbent assay (ELISA) kits for detecting rotaviruses (Abbott Laboratories, North Chicago, Ill., USA) and latex agglutination test kits (Orion Diagnostica, Helsinki, Finland) are commercially available and have recently been evaluated [1-5, 7, 10]. In the present study, four different techniques to detect rotaviruses in feces are compared. Sixty-two fecal specimens were obtained from patients with infantile diarrhea. Specimens were kept at -20~ Samples were suspended in phosphatebuffered solution (PBS), and centrifuged at 3000 rpm for 15rain at room temperature. Supernatant was used for all four techniques. Migration patterns of rotavirus RNA segments by PAGE were determined as described previously [9]. Virus isolation: MA104 cells and primary cynomolgus monkey kidney (PMK) cells were used. The isolation methods have been described previously [6, 8]. Latex agglutination (LA): The Rotalex kit was used according to the manufacturers instructions. The results were read under the inverted microscope (x 40, Chiyoda Kogaku). ELISA was carried out according to the manual of the commercially available Rotazyme kit. Results were read by measuring absorbance at 492nm (Quantum II, Abbott). Of 62 samples, 31 were positive and 10 were negative by all four techniques (Table 1). Thirteen samples were positive by three techniques and negative by one, while five samples were negative by three techniques and positive by one. The results for the 62 samples tested are as follows: 79% (49/62) for PAGE, 65% (40/62) for virus isolation, 73% (45/62) for LA, and 61% (37/62) for ELISA. Several different migration patterns of RNA segments can be ascertained by PAGE [8]. However, there were no distinct correlations between migration patterns and the results obtained in this study. The commercially available

Molecular phylogenetic analysis of Orientia tsutsugamushi based on the groES and groEL genes.

DNA sequences encoding the GroES and GroEL proteins of Orientia tsutsugamushi were amplified by the PCR and sequenced. Pairwise alignment of full-length groES and groEL gene sequences indicated high sequence similarity (90.4-100% and 90.3-100%) in O. tsutsugamushi, suggesting that these genes are good candidates for the molecular diagnosis and phylogenetic analysis of scrub typhus. Comparisons of the 56-kD type-specific antigen (TSA) protein gene and the groES and groEL genes showed that genotypes based on the 56-kD TSA gene were not related to a cluster containing the groES and groEL genes in a dendrogram, suggesting that a gene rearrangement may be associated with homologous recombination in mites.

THE CHANGING SEROEPIDEMIOLOGY OF VARICELLA IN JAPAN 1977-1981 AND 2001-2005

We conducted an anti-varicella-zoster virus antibody seroepidemiologic survey comparing the pre- and postvaccine eras of 1977–1981 and 2001–2005. For each period, 828 samples were measured by enzyme immunoassay test and compared. The differences from 1-year-old to high-school aged children were statistically significant. The introductions of optional varicella immunization and lifestyle changes for children were considered influencing factors.

Diagnostic significance of specific IgA coproconversion in rotavirus infection

early functional abnormalities within a few weeks, can occur at any time after the onset of diabetes, and are reversible by improvement in diabetic control. Later, the metabolic disturbances are followed by structural and irreversible lesions. At this stage, clinical neuropathy is related to the duration of diabetes. So, the suggested explanation for the discrepancies between various studies could be: (1) the number of patients with clinical signs of diabetic neuropathy (higher in Lorinis study); (2) the search for late components indicating low conduction velocity in some nerve fibres, even in patients in whom the PMNCV is normal (not mentioned in Lorinis study). Recently, we have found that the femoral nerve conduction velocity also is unrelated to the duration of diabetes [7]. In contrast to subclinical peripheral neuropathy, we have shown previously that the prevalence of incipient retinopathy, as diagnosed by fluorescein angiography, clearly was related to the duration of diabetes [2]. However, the mechanisms responsible for retinal microangiopathy are different, including changes in the vascular wall as well as haemorrheological perturbations. These microvascular modifications seem to need more time for installation than the neuropathic abnormalities [3]. On the other hand, the breakdown of the bloodretineal barrier may be precocious and is dependent on metabolic control even in short time periods [1, 5]. The discrepancies concerning the relationship between PMNCV and age could be explained by the young age of the patients included in some studies, and may be biased by the duration of diabetes and by metabolic control.