No abstract

A nucleic acid hybridization technique has been developed to study the effect of different antiviral compounds on the replication of human cytomegalovirus in vitro. One laboratory strain of human cytomegalovirus, Ad. 169, and six clinical isolates were studied. Doses needed for 50%6 inhibition of viral DNA replication were calculated fbr foscarnet, acyclovir, and arabinosyladenine. The (8,19,20), acyclovir (ACV) (10, 13, 17), and arabinosyladenine (ara-A) (6). The specificity of most antiviral cotmnds is at the level of the herpesvirus-induced DNA polymerases (9).The effect of antiviral substances is usually studied by determining the reduction in production of infectious virus particles, e.g., the plaque reduction technique or titrations of produced virus particles. Other techniques include enzyme-linked immunosorbent assay for the effect on the synthesis of viral proteins (18)

A prospective study of cytomegalovirus (CMV) infections has been carried out in 28 renal graft recipients. The protocol called for frequent blood and urine sampling during the first year after transplantation, but death or graft loss caused earlier termination in nearly half the patients. In this material 5/7 (71%) susceptible patients developed primary infections and 20/21 experienced a secondary infection (95%). Viruria was detected in 79% and viremia in 43%. The type of blood cell responsible for the viremic phase was studied by separating the blood cells on a density gradient. The polymorphonuclear cell fraction was the most common source of virus but virus could also be recovered from the mononuclear cell fraction. As some samples that were freeze-thawed repeatedly never yielded virus, it would appear that viable cells are needed for virus isolation. In both primary and secondary infections isolation of CMV from blood cells often preceded the isolation of CMV from urine. Among variables tested for a possible relationship to the occurrence of CMV viremia the only one to display such an association was the time at which rejection episodes occurred. In 19/28 such episodes recorded in 19 patients there was a temporal relationship to viremia (p less than 0.03). Seven of the patients experienced clinical symptoms suggestive of CMV infection as fever, cough, myalgia, arthralgia, chest pain and pneumonia. Laboratory signs included elevated amino acid transferase levels, leukopenia and thrombocytopenia and a specific anti-CMV antibody response.

Carcinoembryonic antigen and cross-reacting NCA were demonstrated in 413 amniotic fluid samples representing different stages of pregnancy. Radioimmunoassay of CEA showed a decrease from a mean of 79 ng/ml at 19 weeks to 50 ng/ml near term. NCA, on the other hand, increased slightly from 184 ng/ml at 19 weeks to 219 ng/ml in the terminal part of pregnancy. There was a considerable interindividual variation of both substances at each time period. The NCNCEA quotient increased from 2.5 at 19 weeks to 4.9 at the end of gestation, but the variation precluded its use as a maturation index. The correlations between CEA, NCA, NCNCEA and alpha-fetoprotein levels and the lecithin/ sphingomyelin ratio were studied. The best correlations were found between CEA and NCA levels (r, = 0.52) and between CEA and AFP levels (r, = 0.46). CEA values deviating from the mean 2 2 SD were found in 3 out of 12 samples from fetuses with chromosomal aberrations, and in 3 out of 15 samples from pregnancies with fetuses considered small for date. Two patients with missed abortions both had NCA values outside the normal range. The quotient NCN CEA was lower than expected in two out of four patients with diabetes. Raised AFP levels were found in all ten pregnancies with neural tube defects and in two out of seven patients with chromosomal aberrations.Cancer 42:1579-1584, 1978.ARCINOEMBRYONIC ANTIGEN (CEA), which C is found in and has been characterized from tumors of the gastrointestinal tra~t.~,*,l" has been described to occur in endodermal fetal tissue7 and in amniotic fluid."14 Since a high CEA content can be found in meconiumg and raised values are found in discolored amniotic fluids,5 amniotic CEA may derive mainly from the gut of the fetus. CEA is found in low amounts in the adult gut1' and has been thought to be replaced with nonspecific crossreacting antigen (NCA).13,15 A radioimmunoassay of NCA was therefore used to assess the possible presence of NCA and any variations in this during pregnancy. Alpha-fetoprotein (AFP) has been demonstrated in human fetal liver and yolk sac and is presumably filtered to the amniotic fluid. Raised amniotic AFP is encountered in certain Accepted for publication September 8, 1977. fetal malformations.1~3~21.23.24 In nonpregnant subjects, raised serum values can be seen in tumors of hepatic or germ cell origin.' Fetal proteins are of interest as differentiation products and might therefore be useful as markers of maturity. It was accordingly decided to study the relation between known fetal substances during ontogenesis. Normal values of CEA, NCA and AFP in amniotic fluid at different gestational ages were determined, using specific antisera. Amniotic fluids from abnormal pregnancies were then studied for possible deviations from normal. MATERIALS A N D METHODS Clinical MaterialThe total material consisted of 413 samples collected at the Department of Obstetrics and Gynaecology at the Karolinska Hospital, Stockholm. Of these, 24 were serial samples from 11 patients. Pregnancies were classified as nor...

A new synthesis of 5-(monofluoromethyl)- and 5-(difluoromethyl)-2'-deoxy-2'-fluoro-beta-D-arabinofuranosyluracil (F-FMAU and F2-FMAU) is reported. 3',5'-Di-O-(tert-butyldiphenyl)silylated thymidine or FMAU was photochemically brominated with NBS to the corresponding alpha-monobromide, which was hydrolyzed to the 5-hydroxymethyl derivative. Further oxidation of the latter with MnO2 afforded the 5-formyluracil nucleoside. Treatment of these nucleosides with DAST in CH2Cl2 gave the protected alpha-fluorinated nucleosides. Desiylation with TBAF afforded the desired free nucleosides. Also, 5-(trifluoromethyl)-2'-deoxy-2'-fluoro-beta-D-arabinofuranosyluracil (F3-FMAU) was synthesized by copper-catalyzed trifluoromethylation of 5-iodo-2'-fluoro-ara-U (FIAU). These new nucleosides were studied in comparison with the corresponding 2'-deoxy-erythro-pentofuranosyl derivatives, for their inhibitory activity against cellular thymidylate synthase (TS) and [3H]TdR incorporation into DNA, cytotoxicity against HL-60 cells, and antiviral activity against herpes simplex types 1 and 2 (HSV-1 and -2). F2-TDR and F3-TDR strongly inhibited TS and were also quite cytotoxic and antiherpetic, whereas FTDR was only active in the antiviral assay. In the 2'-fluoroarabino series, fluorine substitution at the alpha-methyl function did not alter significantly the antiherpetic activity. Although FMAU and F-FMAU did not inhibit TS to any significant extent, F2-FMAU and F3-FMAU were weakly inhibitory. The latter nucleosides did not inhibit [3H]TDR incorporation into DNA, while all the other alpha-fluorinated thymine nucleosides inhibited the incorporation of radioactivity of [3H]TDR into DNA to various extents. F2-FMAU and F3-FMAU were about 2 orders of magnitude less cytotoxic against HL-60 cells than were F2-TDR and F3-TDR. The results strongly suggest that in both the 2'-deoxy-2'-fluoroarabino and the 2'-deoxy-erythro-pentofurano series the cytotoxic action of the alpha,alpha-difluoro and alpha,alpha,alpha-trifluoro derivatives may involve the inhibition of TS. The synthesis of [2-14C]F2-FMAU, as an experimental imaging agent, is also described. Unfortunately, the highly selective uptake of the labeled compound within infected brain regions previously noted with [2-14C]FMAU was not detected with the derivative [2-14C]F2-FMAU.