Quantitative evaluation of cytomegalovirus DNA in infantile hepatitis.

We used a PCR method to develop a diagnostic assay for the detection of cytomegalovirus (CMV) DNA in infantile hepatitis, which has been suggested to be associated with CMV infection. CMV DNA was detected in 25 (58.1%) of 43 patients with elevated serum alanine aminotransferase (ALT) levels but no jaundice, and no hepatitis B or C as assessed by conventional PCR. None of the samples from 97 healthy infants tested positive for CMV DNA. We assayed CMV DNA quantitatively in blood using a real-time PCR system that allowed reproducible detection of at least 10 copies of CMV DNA. When 1 microg of DNA from each blood sample was used in this system, a good correlation was obtained between the calculated and measured copy numbers of CMV DNA. This system detected CMV DNA in 29 patients (67.4%) with liver dysfunction. Serial studies in patients with liver dysfunction revealed that CMV DNA copy number decreased, ultimately to below 10, as the ALT levels normalized. In contrast, no CMV DNA copies were detectable by the real-time system in any of the samples from control subjects. These results highlight the usefulness of detecting CMV DNA in the diagnosis of infantile hepatitis and indicate that the real-time quantitative PCR assay may be a valuable tool for monitoring CMV-associated infantile hepatitis.

Use of c-myb antisense oligonucleotides to increase the sensitivity of human colon cancer cells to cisplatin.

Human colon cancer SW480DDP and SW620DDP cells resistant to cisplatin exhibited stronger c-myb gene expression than the parent SW480 and SW620 cells. However, cell growth rates in the cisplatin-resistant cell lines remained similar to those of the parent cells. Antisense oligonucleotides to c-myb inhibited c-myb expression and induced increased sensitivity to cisplatin in SW480DDP and SW620DDP cells, but this did not occur with the control sense oligonucleotides. In contrast, the parent cell lines were not affected by antisense oligonucleotides to c-myb. These results indicate that the c-myb gene in human colon cancer is one of the factors related to cisplatin resistance, and support the need to develop anti-cancer therapeutics based on oncogene-targeted antisense oligonucleotide technology.

Quantitative PCR determination of human cytomegalovirus in blood cells.

We evaluated a rapid and sensitive method to determine human cytomegalovirus (CMV) DNA levels in blood cells using a quantitative polymerase chain reaction (PCR) technique. This method is based on real-time detection of PCR using a dual fluorescence-labeled probe and a sequence detector. Ten copies of CMV DNA were detected, when 1 microg of DNA from blood samples was used with this method, and a good correlation was obtained between increased concentrations of copy numbers calculated and measured copy numbers of CMV DNA (r = 0.999). Forty normal subjects exhibited no copies of CMV DNA. On the other hand, a 6-month-old girl tested positive for increased levels 4 weeks after liver transplant. This method is simple, accurate, and sensitive for the quantitative detection of CMV DNA in vivo, indicating possible applications for the diagnosis and monitoring of CMV infection.

In vitro leukemia cell models of Ara-C resistance.

Cells of the human leukemia line K562 were continuously exposed to cytosine arabinoside (Ara-C) at increasing concentrations for 3 months. The resulting cell line, termed K562/AC, showed 48-fold resistance to Ara-C, compared with the parental K562 cells. The sensitivities of K562/AC to adriamycin (ADR), vincristine (VCR) and etoposide (VP16) were similar to those of parental K562. Gene analysis revealed that this cell line lacked expression of the deoxycytidine kinase (dCK) gene, which was evident in Ara-C-sensitive cells. As in K562 cells, multidrug resistance (MDR-1) and multidrug resistance protein (MRP) genes were not expressed in K562/AC. We also established an in vitro model of Ara-C resistance using phosphorothioate antisense oligonucleotides to dCK (dCK-AS). Treatment of K562 with dCK-AS caused decreased dCK expression and 6- to 10-fold increases in resistance to Ara-C, compared with that in cells treated with sense oligonucleotides to dCK (dCK-S) or in non-transfected cells. The cells described here may contribute to the study of a novel mechanism associated with Ara-C resistance, in which reduced dCK activity may play an important role.

Alcohol dementia and alcohol delirium in aged alcoholics.

In the present study, 126 alcoholics aged 60 years or older were compared with 104 alcoholics aged 35-45 years. No dementia was found in the younger group, whereas 62.7% of the aged patients had dementia; the dementia being irreversible in 32.9% of such patients. Cases of so-called alcohol dementia excluding organic brain diseases accounted for 42.1%. The percentage of aged alcoholics having dementia increased with age, being far beyond the frequency of senile dementia in the general aged. Among various physical complications, hepatic injury and myocardiopathy were more frequent in the aged alcoholics than in general aged people, suggesting that hypertension, myocardiopathy and hepatic injury underlie the manifestation of dementia. There was no case of dementia attributable to the direct effect of alcohol distinctly exceeding the effects of various physical factors. Problem behaviors characteristic of the aged group included 'being soaked in drink' and being inebriated, showing no correlation with the presence or absence of dementia. There was no significant difference in frequency of delirium between the aged group and the younger group. However, in aged alcoholics delirium tended to continue for a longer period during abstinence and was more likely to occur even during non-abstinence. A similar trend was found in aged alcoholics with dementia compared with those without dementia.

[Clear cell sarcoma extending into the inferior vena cava].

A 6-year-old boy who had been diagnosed as having Wilms tumor at the prior hospital was admitted to our hospital for investigation of a right flank mass and macrohematuria in October 1990. Magnetic resonance imaging showed Wilms tumor of the right kidney extending into the inferior vena cava. Right radical nephrectomy, lymphadenectomy and thrombectomy were done on October 30th, 1990. Pathological findings indicated clear cell sarcoma of the kidney. One week after the operation, radiotherapy and chemotherapy with actinomycin D (ACTD), vincristine sulfate (VCR), doxorubicin hydrochloride (DXR) and cyclophosphamide (CPM)) were started according to the protocol by National Wilms Tumor Study (NWTS) III. However, he died of bone metastases and pneumomycosis on October 13th, 1992. Clear cell sarcoma of the kidney (CCSK) is a rare tumor accounting for about 4-5% of Wilms tumors. This is the first report on CCSK with tumor thrombus in Japan.

[Two cases of intraurethral condyloma acuminatum].

The first patient was a 43-year-old man with complaint of urethral bleeding. Two small papillary lesions were found at the fossa navicularis. They were diagnosed as condyloma acuminata and excised. The second patient was a 26-year-old man with complaint of tumors at the prepuce, glans and fossa navicularis. They were diagnosed as condyloma acuminata. Circumcision, electric fulguration and excision of urethral tumor were performed.

In-vitro and in-vivo wear profile of composite resins.

The aim of this study was to evaluate in-vitro wear by two types of slurry of 10 commercial composite resin materials (seven hybrid and three microfilled composites). There was great variation in the in-vitro wear pattern by two types of slurry. The wear rate of microfilled composites was greater than that of hybrid composites, and a negative correlation was observed between wear rate and Knoop hardness values among all materials when hydroxyapatite slurry was used. In contrast, the wear rate of hybrid composites was greater than that of microfilled composites when abraded by green carborundum slurry. These abraded surfaces were compared with SEM micrographs of in-vivo composites surface after 4 years of service. The profile of in-vivo wear surfaces was found to be similar to that of in-vitro wear surfaces abraded with hydroxyapatite slurry.

Streptococcal adherence to uncoated and saliva-coated restoratives.

The adherence of Streptococcus sanguis and S. mutans to seven restoratives in the presence and absence of an artificial salivary pellicle has been investigated. The physiochemical surface characteristics of the bacteria and of the restoratives were also measured, together with the effect of salivary coating of restoratives. Zeta potential of uncoated restoratives showed a positive correlation with the number of adherent S. mutans cells and S. sanguis cells. S. sanguis showed a positive correlation with the hydrophobicity of uncoated restoratives. The surface hydrophobicity was decreased upon saliva coating, whereas the zeta potential was slightly increased. The contact angles or zeta potentials of saliva-coated restoratives were positively correlated with those of uncoated restoratives. The number of adherent cells of both S. sanguis and S. mutans to saliva-coated restoratives were decreased after the coating. Zeta potential of saliva-coated restoratives showed a positive correlation with the numbers of adherent S. mutans cells, suggesting that non-specific electrostatic interactions have an important role in the adherence of this species.

Clinical evaluation of two posterior composite resins retained with bonding agents.

Two hundred thirteen carious cavities were restored with two brands of chemically adhesive posterior composite resins by totally etching both the enamel and dentin walls after removing only the caries detector-stainable tissue and with no mechanical retention form in the cavity preparation. Most cavities were extensive, involving more than two surfaces in 74%. The dentin floor was not covered with cement although a spot lining was placed when the cavity was deep. All restorations were examined, usually after 4 years, by photographs and scanning electron microscope observation of replicas, and by criteria established by the U.S. Public Health Service. The materials and the technique proved to be clinically useful.

In vitro color change of composite-based resins.

The adsorption of staining materials to resin restoratives was considered to be influenced by the physico-chemical properties of the resin-based monomers. To study the effects of the surface characteristics of resins on staining, we prepared five visible-light-cured experimental resins without fillers. Staining of these resins was colorimetrically measured. The staining solutions used were the Oil Orange and the Food Red 3 solutions. With the Oil Orange solution, the materials with higher hydrophobicity showed higher staining. With the Food Red 3 solution, the materials with higher water sorption showed higher staining.

Adherence of streptococci to surface-modified glass.

Four types of surface-modified glass were prepared. Aminopropyl glass was prepared by alkylsilylation of glass slides with gamma-aminopropyltriethoxysilane. This glass carries primary amino groups which may be protonated at pH 7.2. Owing to the presence of both positively charged ions and hydrophobic ethoxyl groups, the glass is considered to be amphipathic. Three other types of surface-modified glass slides were prepared from aminopropyl glass by forming Schiff's bases with three aldehydes: glucose, glyoxylic acid and hexanal. The aldehyde-treated slides were subsequently reduced using sodium borohydride. Thus, the surface of the glass was rendered hydrophilic, ampholytic or hydrophobic, respectively. The adherence of two Streptococcus sanguis strains and two Streptococcus mutans strains to the surface-modified glass slides was studied. Different strains showed differences in adherence to these slides depending on their physico-chemical surface properties. For S. sanguis ATCC 10556, hydrophobic bonds seemed to be most important, while in S. mutans OMZ 176, ionic interactions made the highest contribution to adhesion. Hydrogen bonds seemed to contribute least to adherence.

Streptococcal adherence on various restorative materials.

The adherence of Streptococcus sanguis ATCC 10556, S. sanguis ATCC 10557, S. mutans Ingbritt, and S. mutans OMZ 176 to the surfaces of composite resins, amalgam alloys, and a Au-Ag-Pd alloy was measured. Adhesion was correlated with values for hydrophobicity and zeta-potential of the bacteria and the restorative materials. The hydrophobicity of the restoratives showed a positive correlation between the numbers of adherent S. sanguis cells, suggesting that hydrophobic interactions are important for the adherence of this bacterial species. In contrast, the numbers of adherent S. mutans cells showed a positive correlation with the zeta-potential of the restoratives, suggesting that electrostatic interactions are important in adherence of this bacterium.