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Ethyl 2-alkyl-4-aryl-3-oxobutyrates were synthesized from the corresponding arylacetonitriles and 2-bromo esters. Condensation of the butyrates with thiourea followed by treatment with chloroacetic acid afforded the 5-alkyl-6-(arylmethyl)uracils. Condensation of the uracils with acetals using trimethylsilyl triflate (TMS triflate) as a catalyst gave acyclic 5-alkyl-6-(arylmethyl)uracil derivatives. 6-Benzyl-5-ethyluracil was also condensed with methyl 5-O-(tert-butyldiphenylsilyl)-2-deoxy-3-O-(phenoxythiocarbonyl)-α,β-d-erythro-pentofuranoside, followed by Barton reduction and deprotection, to give the anomers of 6-benzyl-5-ethyl-2‘,3‘-dideoxyuridine. Alkylation of the uracils with alkyl chloromethyl sulfides gave new thio analogues of HEPT. All new N¹-substituted uracils were tested for activity against HIV-1, and the thio analogues were found extremely potent.

Subcellular localization of viral antigens was demonstrated during viral morphogenesis using herpes simplex virus type 1 (HSV-1) infected monolayers of rabbit cornea cells. The localization was done by immunoelectron microscopy employing the peroxidase-antiperoxidase (PAP) immunocytochemical technique and the postembedding staining method. The localization of viral antigens was followed at time intervals during infection from 2 to 19 hr. After exposure of sections to either polyspecific antibodies against total HSV-1 antigens or monospecific antibodies against HSV-1 antigen No. 8, specific immunological reaction products were identified both in the cytoplasm and nucleus after 2 hr. The distribution and quantity of reaction products varied in the infected cells during the viral morphogenesis. The present results on the subcellular distribution of the HSV-1 antigens are related to current biochemical findings.

Fibroblast cultures established from the skin of 56 healthy controls and 15 untreated Stages I and II Hodgkin's patients (HD) were studied in their 3rd, 4th and 5th in vitropassage with respect to transformation with Simian sarcoma virus (SSV) and SV40 and with respect to replication of herpes simplex virus (HSV) Types 1 and 2, pox virus and interferon release. Susceptibility to the 5 viruses varied independently, except for an inverse correlation between susceptibility to SSV and HSV. HD cultures showed a depressed replication of both types of HSV. There was a borderline (P = 0.02) correlation between magnitude of HSV replication and presence of HL-A type B-w44, but this does not explain the HD control difference. Furthermore, the level of serum antibodies to HSV common antigen was not related to magnitude of in vitroreplication. The results thus speak against generally enhanced cellular susceptibility to HSV as a reason for the high titres of serum antibodies to HSV in HD patients.

The nonionic detergent Triton X-100 was used to extract antigens of rubella virus from infected tissue culture cells. Three virus-specific antigens were demonstrated by crossed immunoelectrophoresis by using a pool of human gamma globulin as antiserum. The most dominant of these antigens were purified by ion-exchange chromatography on diethylaminoethyl-cellulose. This antigen was of glucoprotein nature and had slow electrophoretic motility and low binding capacity to diethylaminoethyl-cellulose. Thus, it seems likely that the antigen is identical with the precipitating antigen of rubella virus designated b-antigen or tro-osmophoresis with precipiting antibody in sera obtained from patients recovering from acute postnatal rubella. The precipitin reaction that could be correlated to the hemaglutination-inhibition titers of the same sera appeared 12 days after onset of the disease and remained positive for several years.

Risk factors for genital human papillomavirus (HPV) types 6/11 and 16/18 and any HPV type as well as for herpes simplex virus type 2 (HSV-2) infections were investigated in a population-based study of 1,600 randomly selected women (20-39 years) from Godthåb (native, Nuuk), Greenland, and Nykøbing Falster, Denmark. A total of 586 Greenlandic women and 661 Danish women were included, respectively. They all had a personal interview and a gynecologic examination with cervical smear and swab for HPV analysis (filter in situ hybridization). Moreover, a blood sample was obtained for analysis for HSV-2 antibodies by enzyme-linked immunosorbent assay. In view of the general concept of HPV as a sexually transmitted virus, it is unexpected that women with "multiple" partners revealed a significantly lower risk for all types of HPV than did women with "few" partners, the odds ratio (OR) being 40-60% decreased in women with greater than or equal to 30 partners compared with women who had 0-4 sexual partners. In contrast, the risk for HSV-2 infection was significantly increased among women with early age at first sexual intercourse (OR = 2.9 for age less than or equal to 13 vs. 14-16 years) and multiple partners (OR = 2.6 for greater than or equal to 20 vs. 0-4 partners) (hereafter referred to as "high sexual activity" as well as with an increasing number of sexually active years with an unprotected cervix (i.e., without the use of barrier contraceptives) (OR = 2.0 for greater than or equal to 15 vs. 0-9 years). The results of this study thus demonstrate a surprising risk pattern for HPV types 6/11 and 16/18, but a pattern for HSV-2 in line with that to be expected for a sexually transmitted virus. This could indicate the existence of factors, especially in women with high sexual activity, which interfere with the expression of HPV or with the ability to detect it and/or that HPV may be transmitted by means other than sexual contact.

The nonionic detergent Triton X-100 was capable of solubilizing 90% of the protein content in herpes simplex virus (HSV)-infected rabbit cornea cells. The solubilized HSV antigens formed well-characterized precipitates by crossed immunoelectrophoresis in Triton X-100-containing agarose gel, allowing both identification and relative quantitation. Water-soluble and detergent-requiring HSV antigens were identified by different solubilization procedures in buffer with and without detergent. Five glycoprotein antigens were solubilized only in the presence of detergent, indicating their membrane-bound state. One non-glycosylated antigen was present in both a water-soluble and a membrane-bound form. Based upon the crossed immunoelectrophoretic precipitating patterns of Triton X-100-solubilized HSV antigens, it has been estimated that infected cells yield an amount of virus-specific protein equivalent to 2,000 enveloped virions per cell. Rabbits inoculated intracutaneously with Triton X-100-solubilized HSV antigens developed neutralizing antibodies against HSV almost as effectively as rabbits with an active HSV infection. Precipitins against individual HSV antigens in sera from rabbits infected with HSV and immunized with the Triton X-100-solubilized HSV antigens were assayed by the crossed immunoelectrophoretic technique. Sera from infected rabbits reacted more strongly and with a higher number of HSV antigens than sera from immunized rabbits.

Crossed immunoelectrophoresis was used to characterize herpes simplex virus type 1 (HSV-1) antigens produced by infected HEp-2 cells. We report on a method for analyzing the polypeptide content in individual antigen-antibody precipitates eluted from the second-dimensional agarose gel. Four glycoprotein antigens of HSV-1, Ag-8, Ag-11, Ag-6, and Ag-3, were isolated and analyzed for polypeptide content. The molecular weights of the polypeptides are presented.

In the capture competition immunoassay, undiluted serum was reacted in solution with purified human immunodeficiency virus (HIV) antigen in wells of microtest plates coated with anti-HIV immunoglobulin G antibodies (HIV capture antibodies). HIV antibodies present in the serum being tested combined with the HIV antigen and thus blocked (completely or partially) the fixation of the antigen to the capture layer. Unblocked antigenic activity was measured in subsequent steps by the use of biotinylated anti-HIV immunoglobulin G and peroxidase-conjugated avidin. The assay was evaluated in comparison with indirect enzyme-linked immunosorbent assay and Western (immuno-) blot (WB). A total of 180 serum samples which reacted repeatedly as positive in indirect enzyme-linked immunosorbent assay but negative in WB were found to be negative by the capture competition assay. Of 54 serum samples showing dubious reactions (single p24 bands in WB), 53 were clearly separated into positive or negative reactions, whereas 1 serum sample gave a borderline reaction. It was concluded that a characteristic feature of this kind of inhibition assay is a very low frequency of equivocal results.

A 131,000 molecular weight herpes simplex virus type 1 (HSV-1) glycoprotein designated antigen number 6 (Ag-6) was previously shown to possess almost exclusively HSV-1-specific antigenic sites. Fused rocket and crossed immunoelectrophoresis of fractions obtained from hydroxylapatite chromatography of crude HSV-1 antigen (Triton X-100-solubilized, infected tissue culture cells) showed that a subfraction of Ag-6 could be separated from the other HSV antigens. Enzyme-linked immunosorbent assay with the isolated Ag-6 showed that sera from rabbits infected with HSV-1 and HSV-1 human antisera contained antibodies to Ag-6, whereas sera from HSV-2-infected rabbits and sera from patients with primary HSV-2 infections did not react with Ag-6. Enzyme-linked immunosorbent assay of 852 human sera for antibodies to HSV type-common glycoproteins, Ag-6, and HSV 2-specific antigens showed that 139 sera which reacted negatively with HSV type-common glycoproteins also did not react with Ag-6 with HSV-2 specific antigens. The 713 sera reacting positively to HSV type-common antigens either reacted with Ag-6 (328 sera) or with HSV-2-specific antigens (31 sera) or both (354 sera). This means that Ag-6 might be useful in large-scale human serology for the detection of past infection with HSV-1, irrespective of whether or not past infection with HSV-2 has occurred.

Nomograms or simple formulas have been suggested for estimating the glomerular filtration rate (GFR) from either plasma creatinine (ClK) or from a single plasma sample drawn after an intravenous injection of 51Cr-EDTA (ClA/B)- This study compares the precision of estimating GFR as ClK and ClA/B-A multiple plasma-sample standard method (ClSM) served as reference. The comparison was performed in 136 individuals. The variance of ClAJB on ClSM was 14.4 ml/min/1.73 m2 while the variance of C1K on ClSM was 204.5 ml/min/1.73 m2. It is concluded that the ClA/B values are more precise than the C1K values (p<0.01). The single-sample method for determination of 51Cr-EDTA clearance seems well-suited for evaluation and monitoring of patients with known or suspected kidney diseases. It may even be applied for screening purposes in cases where plasma creatinine would otherwise have been chosen instead of the usual resource-demanding method for determination of C1SM.

By use of crossed immunoelectrophoresis with intermediate gel, antidbody titers against six individual herpes simplex virus (HSV) glycoproteins and two nonglycosylated proteins were determined in 100 human sera. High antibody titers were found against two different HSV type-common glycoproteins designated Ag8 and Ag11 (containing glycosylated polypeptides D and B, respectively). The anti-Ag8 and -Ag11 titers correlated with HSV neutralizing antibody titers. Most of the serological cross-reactivity between HSV type 1 and type 2 was probably caused by antibodies to Ag8 and Ag11. Human antibodies against one HSV type 1-specific glycoprotein (Ag6, containing glycosylated polypeptide C) and two HSV type 2 glycoproteins (Ag4 and Ag9) were also demonstrated, and the titers correlated better with neutralizing antibody titers of the homologous than of the heterologous virus type. The data presented can be directly applied to the further development of diagnostic reagents.

Twenty immunocompetent patients, four females and 16 males, with severe recurrent genital herpes (median number of recurrences the previous year 16, range (8-24] entered an open continuous long-term suppressive treatment with oral acyclovir (ACV) for 12 months. The study included a dose-titration schedule: (ACV, 200 mg x 4/1-3 months, ACV, 400 mg x 2/4-6 months, ACV, 200 mg x 2/7-9 months, and ACV, 400 mg x 1/10-12 months). Patients with recurrences on steps two and three received an alternative dose of ACV, 200 mg x 3. Otherwise patients entered the previous dose-step. Five (20%) of patients were completely free of symptoms (recurrences and abortive lesions) during the four dose-reduction periods. A further nine patients (50%) could be dose-reduced to 200 mg x 3 without symptoms. Isolates from three patients showed a decrease in virus sensitivity after ceasing treatment. In conclusion, 14/20 of treated patients could be dose reduced to 200 mg x 2-3 without selection of HSV strains showing clinically important decreases in sensitivity towards ACV. [ABSTRACT FROM PUBLISHER]