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Herpes zoster (HZ) results from the reactivation, multiplication, and spread of varicella-zoster virus (VZV) that remains latent in sensory neurons following earlier primary VZV infection (ie, varicella or chickenpox) [1]. Department of Veterans Affairs (VA) Cooperative Study 403: The Shingles Prevention Study was a randomized, double-blind, placebo-controlled efficacy trial of live attenuated Oka/Merck HZ vaccine (zoster vaccine) in 38 546 adults ≥60 years of age. The Shingles Prevention Study demonstrated that zoster vaccine reduced the HZ burden of illness by 61.1% (95% confidence interval [CI], 51.1–69.1), the incidence of postherpetic neuralgia (PHN) by 66.5% (95% CI, 47.5–79.2), and the incidence of HZ by 51.3% (95% CI, 44.2–57.6) through 4 years of postvaccination follow-up [2–4]. After the completion of the Shingles Prevention Study, a cohort of subjects was re-enrolled into a short-term persistence substudy. The Short-Term Persistence Substudy extended the follow-up of this cohort of subjects to collect data on the persistence of zoster vaccine efficacy for the 3 end points described above during the interval between the closeout of VA Cooperative Study 403 and the completion of the final data analysis. The Short-Term Persistence Substudy also provided a cohort of the original Shingles Prevention Study vaccine recipients who could subsequently be enrolled in a long-term persistence substudy to further assess the persistence of vaccine efficacy. In addition, data on the persistence of vaccine efficacy for each year after vaccination in the Shingles Prevention Study have not been published. Therefore, the objectives of this article are to assess the persistence of vaccine efficacy for the 3 study end points in the Short-Term Persistence Substudy population, the Shingles Prevention Study population, and the combined Shingles Prevention Study and Short-Term Persistence Substudy populations and to assess the persistence of vaccine efficacy for the 3 study end points for each year through year 7 after subjects received zoster vaccine or placebo in the Shingles Prevention Study.

In patients with chronic hepatitis B, brief lamivudine therapy suppresses hepatitis B virus (HBV) DNA but results infrequently in sustained losses of virus replication posttreatment. We evaluated treatment response and its posttreatment durability during up to 18 months of lamivudine therapy (100 mg/d) in 24 patients who had hepatitis B e antigen (HBeAg) despite 1 to 3 months of prior therapy. Therapy was to be stopped after HBeAg loss or seroconversion (acquisition of antibody to HBeAg); posttreatment monitoring continued for 6 months. During therapy, which was well tolerated, HBV DNA became undetectable in all evaluable patients, accompanied by reduced alanine transaminase (ALT) activity. The cumulative 18-month confirmed loss of HBeAg during therapy was 9 of 24 (38%) and seroconversion was 5 of 24 (21%). Therapy was discontinued after HBeAg loss/seroconversion in 7 patients, and HBeAg status was maintained in all. Four of the patients with HBeAg responses lost HBsAg at least once. In 10 (43%) of 23 patients tested, we identified HBV polymerase YMDD mutations, 3 with detectable HBV DNA (2 with ALT elevations) and 7 without virological/biochemical breakthrough. In conclusion, up to 18 months of lamivudine therapy was well tolerated, suppressed HBV replication consistently, and tripled the frequency of HBeAg losses observed during brief-duration therapy; HBeAg loss/seroconversion remained durable posttreatment. The emergence of YMDD-variant HBV was relatively common but occurred typically without reappearance of detectable HBV DNA or ALT elevation. Our observations suggest that lamivudine can be stopped after confirmed HBeAg loss or seroconversion.

Members of the herpes virus family and hepatitis B virus (HBV) have been implicated as etiologic agents in non-A, non-B (NANB) fulminant hepatic failure (FHF), but the frequency of infection with these agents has not been established using appropriate controls. To examine this issue, we studied 50 NANB FHF patients and 104 liver transplant recipients from North America and Europe. Hepatic DNA was analyzed by polymerase chain reaction (PCR) for evidence of Epstein-Barr virus (EBV), cytomegalovirus (CMV), herpes simplex virus I (HSV I) and II (HSV II), varicella-zoster virus (VZV), and human herpes virus-6 (HHV-6) nucleic acid sequences. The prevalence of HBV was assessed in North American subjects only. HSV I, HSV II, VZV, and HHV-6 viral sequences were not observed in any samples. Three of 50 FHF (6%) and 14 of 104 control patients (13%) were positive for CMV DNA. Two of 50 FHF (4%) and 10 of 104 control patients (10%) had EBV DNA, and HBV DNA was observed in 3 of 10 North American FHF patients (30%) and 3 of 59 controls (5%) without serum markers for HBV infection. The finding of HBV DNA in the liver of seronegative controls from North America but not Europe suggests that occult hepatitis B sequences in patients with NANB FHF may simply reflect geographic differences. The majority of cryptogenic FHF cases cannot be attributed to infection with herpes viruses or HBV.

Background The herpes zoster vaccine is effective in preventing herpes zoster and postherpetic neuralgia in immunocompetent older adults. However, its safety has not been described in depth. Objective To describe local adverse effects and short- and long-term safety profiles of herpes zoster vaccine in immunocompetent older adults. Design Randomized, placebo-controlled trial with enrollment from November 1998 to September 2001 and follow-up through April 2004 (mean, 3.4 years). A Veterans Affairs Coordinating Center generated the permutated block randomization scheme, which was stratified by site and age. Participants and follow-up study personnel were blinded to treatment assignments. (ClinicalTrials.gov registration number: NCT00007501) Setting 22 U.S. academic centers. Participants 38 546 immunocompetent adults 60 years or older, including 6616 who participated in an adverse events substudy. Intervention Single dose of herpes zoster vaccine or placebo. Measurements Serious adverse events and rashes in all participants and inoculation-site events in substudy participants during the first 42 days after inoculation. Thereafter, vaccination-related serious adverse events and deaths were monitored in all participants, and hospitalizations were monitored in substudy participants. Results After inoculation, 255 (1.4%) vaccine recipients and 254 (1.4%) placebo recipients reported serious adverse events. Local inoculation-site side effects were reported by 1604 (48%) vaccine recipients and 539 (16%) placebo recipients in the substudy. A total of 977 (56.6%) of the vaccine recipients reporting local side effects were aged 60 to 69 years, and 627 (39.2%) were older than 70 years. After inoculation, herpes zoster occurred in 7 vaccine recipients versus 24 placebo recipients. Long-term follow-up (mean, 3.39 years) showed that rates of hospitalization or death did not differ between vaccine and placebo recipients. Limitations Participants in the substudy were not randomly selected. Confirmation of reported serious adverse events with medical record data was not always obtained. Conclusion Herpes zoster vaccine is well tolerated in older, immunocompetent adults. Primary funding source Cooperative Studies Program, Department of Veterans Affairs, Office of Research and Development; grants from Merck to the Veterans Affairs Cooperative Studies Program; and the James R. and Jesse V. Scott Fund for Shingles Research.

The results of the Shingles Prevention Study, a Cooperative Studies Program conducted through the US Department of Veterans Affairs, are summarized. Also provided are general recommendations and contraindications for the use of zoster vaccine live among patients aged 60 years or older.

A real-time PCR assay was developed to identify varicella-zoster virus (VZV) and herpes simplex virus (HSV) DNA in clinical specimens from subjects with suspected herpes zoster (HZ; shingles). Three sets of primers and probes were used in separate PCR reactions to detect and discriminate among wild-type VZV (VZV-WT), Oka vaccine strain VZV (VZV-Oka), and HSV DNA, and the reaction for each virus DNA was multiplexed with primers and probe specific for the human beta-globin gene to assess specimen adequacy. Discrimination of all VZV-WT strains, including Japanese isolates and the Oka parent strain, from VZV-Oka was based upon a single nucleotide polymorphism at position 106262 in ORF 62, resulting in preferential amplification by the homologous primer pair. The assay was highly sensitive and specific for the target virus DNA, and no cross-reactions were detected with any other infectious agent. With the PCR assay as the gold standard, the sensitivity of virus culture was 53% for VZV and 77% for HSV. There was 92% agreement between the clinical diagnosis of HZ by the Clinical Evaluation Committee and the PCR assay results.

In May 2008, the Advisory Committee on Immunization Practices (ACIP) of the Centers for Disease Control and Prevention in the USA recommended the routine administration of a live-attenuated high-potency varicella zoster vaccine to all adults over 60 years of age without specific contraindications, for the prevention and attenuation of herpes zoster (HZ). Nevertheless, many physicians still consider this vaccine to be of marginal value. This is not a reasonable conclusion. This short article reviews available data.

In this 3-year study of suppressive acyclovir for recurrent genital herpes, patients with more than six recurrences per year were randomized initially to 400 mg of acyclovir or placebo orally two times per day, with recurrences treated with 200 mg of acyclovir five times per day for 5 days. In the second year of the study, all patients received acyclovir as a daily suppressive or intermittent acute therapy; in the third year, all received daily acyclovir. Among 525 patients completing 3 study years, 289 received 3 years of suppressive therapy and 236 received 1 year of acute therapy followed by 2 years of suppressive therapy. Of those who completed the third year, 61% were recurrence free that year; 25% of the suppressive therapy-only group were recurrence free for all 3 years. The annual recurrence rate dropped from more than 12 recurrences per year at baseline to 1.0 (suppressive therapy) and 1.4 (acute and suppressive therapy) recurrences during the third year. No significant toxic effects were detected. Daily suppressive acyclovir therapy was effective and well tolerated.

Topic The role of the zoster vaccine in the prevention of herpes zoster and its sequelae, including postherpetic neuralgia (PHN) and herpes zoster ophthalmicus. Clinical Relevance Wide administration of the herpes zoster vaccine in accordance with the recommendations of the Centers for Disease Control and Prevention Advisory Committee on Immunization Practices (ACIP) will lead to a decline in the incidence and morbidity of herpes zoster and its complications, including PHN. Methods The key study leading to the approval of the zoster vaccine for use, the Centers for Disease Control and Prevention ACIP's recommendations for appropriate use of the zoster vaccine, and predictions regarding the cost efficacy of a zoster vaccination program are reviewed. Results The Shingles Prevention Study established that the zoster vaccine was safe, well tolerated, and effective in reducing the burden of illness due to herpes zoster and the incidence of PHN. The ACIP recommended that the zoster vaccine be given to adults 60 and older for the prevention of herpes zoster. Cost-efficacy analyses suggest that the greatest gain in quality-adjusted life-years can be gained by vaccinating individuals at the younger end of the ACIP-recommended age range. Conclusion The zoster vaccine promises to reduce the morbidity and mortality of herpes zoster. Administering the vaccine at the younger end of the age range may offer a greater cost benefit.

background The incidence and severity of herpes zoster and postherpetic neuralgia increase with age in association with a progressive decline in cell-mediated immunity to varicella– zoster virus (VZV). We tested the hypothesis that vaccination against VZV would decrease the incidence, severity, or both of herpes zoster and postherpetic neuralgia among older adults. methods We enrolled 38,546 adults 60 years of age or older in a randomized, double-blind, placebo-controlled trial of an investigational live attenuated Oka/Merck VZV vaccine (“zoster vaccine”). Herpes zoster was diagnosed according to clinical and laboratory criteria. The pain and discomfort associated with herpes zoster were measured repeatedly for six months. The primary end point was the burden of illness due to herpes zoster, a measure affected by the incidence, severity, and duration of the associated pain and discomfort. The secondary end point was the incidence of postherpetic neuralgia. results More than 95 percent of the subjects continued in the study to its completion, with a median of 3.12 years of surveillance for herpes zoster. A total of 957 confirmed cases of herpes zoster (315 among vaccine recipients and 642 among placebo recipients) and 107 cases of postherpetic neuralgia (27 among vaccine recipients and 80 among placebo recipients) were included in the efficacy analysis. The use of the zoster vaccine reduced the burden of illness due to herpes zoster by 61.1 percent (P

Purpose: The authors describe an immunocompetent patient who developed multiple recurrent branch retinal artery occlusions (BRAOs) associated with the varicella zoster virus (VZV). Methods: A 69-year-old woman with mild bilateral vitritis developed superior and inferior BRAOs in her right eye with decreased visual acuity to 20/40, and a peripheral BRAO inferotemporally in her left eye. One month later, the inferotemporal BRAO progressed proximally in her left eye with a decrease in visual acuity to 20/40. After an extensive negative systemic evaluation, she underwent a diagnostic pars plana vitrectomy of her right eye. Results: Vitreous fluid was positive for VZV DNA by polymerase chain reaction (PCR). The patient was treated with intravenous acyclovir and systemic oral steroids. After remaining disease free for 3 months, the patient had two recurrences: 1) a mild vitritis and 2) development of a new superior temporal artery occlusion in the left eye. Both recurrences were treated with oral acyclovir and systemic steroids. The patient remained recurrence free for 12 months on a maintenance dose of oral acyclovir, and for 4 additional months without acyclovir. Conclusions: Varicella zoster virus can be associated with the syndrome of multiple recurrent BRAOs. The diagnosis of VZV-associated BRAO can be established by PCR of intraocular fluid.

A live attenuated varicella-zoster virus (VZV) vaccine was developed by Takahashi and his colleagues in the early 1970s (1). The vaccine has been designated the Oka strain after the child from whom the virus was isolated. Attenuation was believed to have been induced by serial passage of the original clinical isolate at various temperatures and in different cell lines (1, 2). Numerous studies throughout the world have subsequently shown the vaccine to be safe, immunogenic, and highly protective in both healthy and immunosuppressed individuals (3–6).

Background: We conducted a trial to compare treatment with zidovudine or didanosine in patients with advanced human immunodeficiency virus type 1 (HIV-1) infection who had received little or no previous therapy with zidovudine. Methods: Six hundred seventeen patients with acquired immunodeficiency syndrome (AIDS), advanced AIDS-related complex (CD4 cell count, ≤0.30× 10 9 /L [300/μL]), or asymptomatic HIV (CD4 cell count, ≤0.20× 10 9 /L) received zidovudine, 500 mg/d of didanosine, or 750 mg/d of didanosine in a randomized, double-blind allocation, with cross-over to alternative medication after development of an end point or serious toxic effect. To be eligible, patients must have received either no or up to 16 weeks of zidovudine therapy before entry into the study. Primary end points were development of a new AIDS-defining event or death. Secondary clinical end points were new or recurrent AIDS-defining events, or death, and survival. Results: In the study as a whole, there were no differences in the relative risks (RRs) of the development of end points between treatment groups. However, there was a strong interaction between the relative efficacies of zidovudine and didanosine and previous experience with zidovudine. Among 380 patients with no previous zidovudine therapy, zidovudine was more effective than 750 mg/d of didanosine (RR, 1.43; 90% confidence interval [CI], 1.02 to 2.00), with a similar trend for zidovudine compared with 500 mg/d of didanosine (RR, 1.21; 90% CI, 0.86 to 1.71). However, among 118 patients with more than 8 weeks but no more than 16 weeks of previous zidovudine therapy, 500 mg/d of didanosine was more effective than zidovudine (RR, 0.48; 90% CI, 0.27 to 0.86); there was a similar trend for increased effectiveness of 750 mg/d of didanosine compared with zidovudine (RR, 0.61; 90% CI, 0.36 to 1.03). Among 119 patients who had some but no more than 8 weeks of previous zidovudine therapy, there were no significant differences among the treatment arms. Similar findings were noted in the analysis of the two secondary clinical end points. No significant differences were found in efficacy between the groups receiving 500 and 750 mg/d of didanosine. The major toxic effect associated with zidovudine was hematopoietic (granulocytopenia) and that associated with didanosine was pancreatitis (dosage, 750 mg/d). Conclusions: In patients with advanced HIV disease, zidovudine appears to be more effective than didanosine as initial therapy; however, some patients with advanced HIV disease may benefit from a change to didanosine therapy after as little as 8 to 16 weeks of therapy with zidovudine. (Arch Intern Med. 1995;155:961-974)

In this 3-year study of suppressive acyclovir for recurrent genital herpes, patients with more than six recurrences per year were randomized initially to 400 mg of acyclovir or placebo orally two times per day, with recurrences treated with 200 mg of acyclovir five times per day for 5 days. In the second year of the study, all patients received acyclovir as a daily suppressive or intermittent acute therapy; in the third year, all received daily acyclovir. Among 525 patients completing 3 study years, 289 received 3 years of suppressive therapy and 236 received 1 year of acute therapy followed by 2 years of suppressive therapy. Of those who completed the third year, 61% were recurrence free that year; 25% of the suppressive therapy—only group were recurrence free for all 3 years. The annual recurrence rate dropped from more than 12 recurrences per year at baseline to 1.0 (suppressive therapy) and 1.4 (acute and suppressive therapy) recurrences during the third year. No significant toxic effects were detected. Daily suppressive acyclovir therapy was effective and well tolerated. ( JAMA . 1991;265:747-751)

Summary Varicella-zoster virus infection of primary hamster embryo cells resulted in oncogenic transformation. These transformed cells exhibited virus-specific antigens by immunofluorescence and developed surface Fc receptors. They induced aggressive fibrosarcomas when injected into inbred hamsters. The tumour-bearing hamsters develop antibodies to varicella-zoster antigens.

Eight patients became clinically reinfected with varicella-zoster virus despite the presence of specific antibody in the blood three days to six months before the onset of illness. One patient had had varicella previously; a second had been immunized with live, attenuated varicella vaccine 10 months earlier. While it was suspected that these patients experienced a reactivation of latent virus that caused atypical disseminated zoster rather than varicella, detailed study of the vaccinated child suggests that this was not the case; by restriction-endonuclease techniques, this vaccinee was shown to have been infected with wild-type varicella-zoster virus despite the presence of specific antibody and cellular immunity to the virus. All cases clinically resembled chickenpox. Thus, not only subclinical varicella (manifested by a rise in antibody titer after close exposure) but also clinical reinfection with the virus can occur. Clinical reinfection probably develops more frequently in immunocompromised than in immunocompetent individuals. Reinfections are usually mild.

The DNA from several clinical isolates of varicella-zoster virus (VZV) were compared with the DNA from the vaccine strain VZV using three restriction endonucleases: BamHI, BgII, and HpaI. When electrophoresed through an agarose gel, the vaccine DNA digestion pattern was significantly different from the digestion patterns of the wild-type DNAs. Variations in the digestion pattern of the separate clinical isolates were also observed.

To determine if the presence of hepatitis B e antigen (HBeAg) and elevated DNA polymerase activity in the serum of chronic HBeAg carriers indicate increased contagiousness in a household setting, the household contacts of 74 carriers were prospectively evaluated for serologic evidence of hepatitis B infection. Thirty of the HBsAg carriers had HBeAg and 44 had anti-HBe. Twenty-eight HBeAg-positive carriers regularly demonstrated elevated DNA polymerase when serially drawn serum samples were analyzed. None of the anti-HBE-positive carriers demonstrated elevation of DNA polymerase activity. Both household contacts of HBeAg-positive and anti-HBe-positive carriers demonstrated serologic evidence of hepatitis B infection (HBsAg, anti-HBs, and anti-HBc). However, infection was significantly more frequent among spouses and sexual partners of carriers who had either HBeAg (P less than 0.001) or elevated DNA polymerase activity (P less than 0.001). Thus, the data indicate that a particular subpopulation of spouses and sexual partners of hepatitis B carriers are at significantly greater risk for acquiring infection.

A mixture of varicella-zoster virus-infected human embryonic lung fibroblasts and hamster embryo cells produced foci of morphologically transformed cells after several weeks of incubation. These transformed cells exhibited virus-specific antigens by immunofluorescence and developed surface Fc receptors. They induced aggressive fibrosarcomas when injected back into inbred hamsters. Cells derived from hamster tumor tissue exhibited similar properties. The tumor-bearing hamsters develop antibodies specific for varicella-zoster virus (VZV) antigens. Cell lines derived from both the original transformants and explanted hamster tumor tissue have varying growth properties. All maintain indefinite growth. All eventually lose varicella-zoster virus-specific immunofluorescence. None retain VZV-specific DNA sequences as determined by dot blot and Southern blot hybridization using radiolabeled whole VZV DNA and cloned VZV DNA fragments as a probe. A few transformed and tumor cell lines frozen relatively soon after isolation and stored in liquid nitrogen were also thawed, replicated to mass culture, and analyzed for VZV-specific DNA sequences. Hybridization with radiolabeled whole VZV DNA initially suggested that some of these cell lines did contain virus-specific DNA sequences. However, hybridization with cloned VZV DNA fragments radiolabeled in vitro and representing greater than 95% of the virus genome was negative. Karyotyping of these "positive" transformed cells indicated that they are of human and not hamster origin. The positive hybridization with whole VZV DNA therefore most likely represented contamination of the probe by host DNA sequences. The cells that survived freezing then were predominantly transformants of human origin. Attempts to repeat the transformation of hamster embryo and baby hamster kidney cells with laboratory-passaged VZV strains have been unsuccessful. Similarly, we have been unable to transform cells with whole VZV DNA or cloned VZV DNA fragments, although whole VZV DNA is demonstrably infectious. Apparently, transformation of cells by the varicella-zoster virus is a very rare event and one that may require a recent clinical isolate. Fresh clinical isolates of varicella-zoster virus are seemingly able to transform mammalian cells in vitro. The transformed cells have malignant properties and are capable of indefinite growth. Although VZV gene function can be detected early after transformation, there is no evidence that a VZV-specific protein product is required. Transformation of mammalian cells by varicella-zoster virus apparently occurs through a "hit-and-run" mechanism.

Partially purified hepatitis B e antigen (HBeAG) was prepared by ultracentrifugation, ammonium sulfate precipitation, and molecular sieve chromatography of sera obtained from asymptomatic carriers of hepatitis B surface antigen. The antigenic specificity of the HBeAG preparations was investigated further with affinity chromatography. The results indicated that HBeAG is distinct and separable from DNA polymerase activity. Columns coupled with either goat IgG prepared from antiserum to human IgG or antibody to HBeAg bound all detectable HBeAg and bound 31% and 100% of the IgG, respectively, from a partially purified HBeAg preparation. Rate zonal sucrose sedimentation and molecular sieve and ion-exchange chromatography indicated a variability in molecular weight and charge; this finding suggested a heterogenous population of immunoreactivities containing HBeAg. Our preliminary results suggest the existence of an HBeAg-IgG complex.

Live attenuated varicella vaccine was administered to healthy varicella-susceptible adults. Of 187 adults immunized with the Oka strain of vaccine, seroconversion to varicella-zoster virus (VZV) occurred in 82% after one dose and in 94% after two doses. Adverse effects were unusual. After immunization, one subject developed mild zoster caused by wild-type virus. Twelve adults developed a mild breakthrough case of chickenpox after exposure to VZV. Protection after household exposure was observed in nine (56%) of 16; however, the illness in all seven patients with breakthrough illness was modified, with an average of only 24 vesicles. Subjects seropositive at household exposure were unlikely to develop a breakthrough illness. Approximately 25% of vaccinees who seroconverted lost detectable antibodies to VZV after vaccination, but even those who became seronegative were partially protected. Varicella vaccine offered significant protection against severe chickenpox in healthy adults.

Two discrete classes of particles appear following the infection of primary African green monkey kidney cells with high multiplicities of an SV40-like isolate from a patient with progressive multifocal leukoencephalopathy (PML). The DNA purified from the nondefective “heavy” particles was very similar to standard SV40 DNA. “Light” viral DNA was circular, supercoiled, 80% the physical size of SV40, and did not contain the Eco RI restriction endonuclease site. The kinetics of “light” viral DNA reassociation indicated a physical complexity equivalent to 40–50% of SV40 DNA; no host sequences were detected in “light” viral DNA. Digestion of “light” viral DNA with the Hemophilus influenzae restriction endonuclease resulted in 5 cleavage products none of which corresponded to the 11 fragments obtained with SV40 DNA.

Restriction endonuclease analysis of varicella-zoster virus (VZV) DNA has been used in unraveling the complex epidemiology of VZV infections in individuals immunized with a live, attenuated varicella virus vaccine. Early rashes appearing within the first six weeks after vaccination are invariably due to vaccine virus. True breakthrough infections with wild-type VZV also occur in vaccinees. Five cases of zoster have been seen in leukemic children vaccinated while in remission. One case appeared 22 months after vaccination in the same general area as the inoculation. The virus isolated was vaccine derived. A second case of zoster appeared in a dermatome unrelated to the sites of vaccination approximately 19 months after apparently natural varicella. This virus was wild type. Vaccine virus can therefore establish latency and can later reactivate as herpes zoster.

Discordant results for e antigen, DNA polymerase activity, and Dane particle frequency were noted in the sera of two patients representing an index case-contact case pair with chronic aggressive hepatitis B. The lack of correlation between presence of e antigen and the infecting viral strain, as well as the strong relationship of e antigen to persistent and significant viremia, emphasize the role of the host's immune response in the expression of this antigen.

Summary Aldoses, but not their polyhydroxyl counterparts, were active in inhibiting enterovirus HA. The fewer the number of carbon atoms in the aliphatic chain, the broader (in terms of the number of enteroviruses which are inhibited from HA) the blocking capacity of the compound. Thus glycerylaldehyde was much less specific and more active than D-galactose, D-mannose or D-ribose. Formaldehyde inhibited the HA of all of the viruses which were tested, namely echoviruses type 3, 7, 11, 12 and 19; coxsackieviruses group B, type 1 and 5; and reovirus type 2. The carbonyl group of these aldoses is thus implicated as the necessary reactive site in these blocking reactions. A number of L-amino acids were tested similarly for their ability to block enterovirus-red blood cell union. They were remarkably inactive. Pretreatment of erythrocytes with D, L-glycerylaldehyde rendered them much less agglutinable by echoviruses, type 3, 11 and 19; reovirus type 2; and coxsackievirus group B, type 1. Similarly treated red blood cells were not altered in their agglutinability by echoviruses type 7 and 12. The active aldoses attach firmly to erythrocytes, but not to virus. Enterovirus HA may involve the binding of a carbonyl group of a terminal sugar from an oligosaccharide side chain on the virus capsid to a specific receptor on the red cell surface.

The location of integrated SV40 DNA in a series of virus-free transformed murine cells was analyzed by characterizing the reiteration frequency of adjacent host cell DNA sequences. Cellular DNA was mechanically sheared to fragments approximately 8 × 10 6 or 7 × 10 5 daltons in size, separated into reiterated and nonreiterated fractions, and then examined for the presence of SV40 DNA. SV40 DNA sequences were associated with the nonreiterated fraction of mammalian DNA in each of the cell lines examined. In addition, the pattern of viral DNA distribution between the repetitive and unique cellular DNA fractions varied from cell line to cell line suggesting that integration of SV40 DNA may occur at different sites in each cell line examined.

RNA type C viruses have been isolated from many animal species. In the mouse and chicken, they exist in an unexpressed form in every cell1–3. Whether endogenous type C viral genetic information is integrated within the cell genome has been studied genetically and biochemically. Loci for inducibility of biologically distinguishable viruses have been detected in mouse cells, suggesting that the loci may represent virus structural information4,5. Numerous biochemical studies have characterized and quantified type C virus-specific DNA in avian and murine cells6–9. In some reports, the number of viral DNA equivalents per cell was quantified by measuring the effect of unlabelled cellular DNA on the reassociation kinetics of a labelled DNA probe10. Using labelled double stranded (ds) DNA prepared in vitro with the reverse transcriptases of RNA tumour viruses, earlier reports indicated that the major portion of the type C virus ds DNA product was complementary to a relatively small fraction of the viral genome7,11; this in vitro DNA probe did not effectively discriminate between the DNAs of different species with respect to the presence of viral genetic information7,8. A portion of the ds DNA product was also shown to have a genetic complexity more representative of the 70S genome7,11.

evaluation method for cerebral palsy. Am J Occup Ther 9:105, 1955. 4. Levine MA, Downs RW, Moses AM, Breslau NA, Marx S J, Lasker RD, Rizzoli RE, Aurbach GD, Spiegel AM: Resistance to multiple hormones in patients with pseudohypoparathyroidism: Association with deficient activity of guanine nucleotide regulatory protein. Am J Med 74:545, 1982. 5. Glinder EM, Heth DA: Colorimetric determination with bound "calmagite" of magnesium in human blood [abstract] Clin Chem 17:662, 1975. 6. Marx S J, Sharp ME, Krudy A, Rosenblatt M, Mallette LE: Radioimmunoassay for the middle region of human parathyroid hormone: Studies with a radioiodinated synthetic peptide. J Clin Endocrinol Metab 53:76, 1981. 7. Chase LR, Melsor GL, Aurbach GD: Pseudohypoparathyroidism: Defective excretion of 3 ' 5 ' -AMP in response to parathyroid hormone. J Clin Invest 48:1832, 1969. 8. Eisman JA, Hamstra A J, Kream BE, DeLuca HF: A sensitive, precise and convenient method for determination of 1-25-(OH)2 D in human plasma. Arch Biochem Biophys 176:235, 1976. 9. Albright F: Hypoparathyroidism as a cause of osteomalacia. J Clin Endocrinol Metab 16:419, 1956. 10. Drezrter MR, Neelon FA, Jowsey J, Lebovitz HE: Hypoparathyroidism: A possible cause of osteomalacia. J Clin Endocrinol Metab 45:114, 1977. 11. Omel'chenko LI, Rodionov VP, Skrypnychencko LS: Pseudo vitamin D deficient rachitis in a child with hypoparathyroidism. Pediatr Akush Ginekol 6:28, 1978. 12. Marx S J: Hereditary resistance to 1,25-dihydroxyvitamin D. Rec Prog Horm Res 40:589, 1984. 13. Liberman VA, Samuel R, Halabe A, Kauli R, Edelstein S, Weisman Y, Papapoulos SE, Clemens TL, Fraher L J, O'Riordan JLH: End organ resistance to 1,25-dihydroxycholecaciferol. Lancet 1:504, 1980. 14. Markowitz ME, Rosen JF, Smith C, De Luca HF: 1,25Dihydroxyvitamin D3-treated hypoparathyroidism: 35 patient years in 10 children. J Clin Endocrinal Metab 55:727, 1982. 15. Pettifor JM, Ross FP, Travers R, Glorieux FH, DeLuca HF: Dietary calcium deficiency: A syndrome associated with bone deformities and elevated serum 1-25 dihydroxy vitamin D concentrations. Metab Bone Dis Rel Res 2:301, 1981. 16. Liebross BA, Coburn JW: Renal osteodystrophy. In Heath D, Marx S J, editors: Clinical endocrinology, vol 2. Calcium disorders. London, 1982, Butterworth Scientific.

Samples of serum inoculated with Escherichia coli and serum that became contaminated with bacteria after exposure to a laboratory atmosphere demonstrated elevated DNA polymerase activity. The levels of activity were well within the range of values found in hepatitis B e antigen (HBeAg)-positive samples. The bacterial polymerase activity was markedly reduced by a single passage of serum samples through a 0.22-micron Millipore filter prior to analysis. Repeated filtration did not result in a substantial further decrease in polymerase activity. In sera that were heavily contamined with E. coli, however, filtration was not successful in reducing bacteria-associated polymerase activity to a base-line uncontaminated level. In such instances double antibody immunoprecipitation proved effective in elimination of bacterial activity. When bacterial contamination of serum samples is a possibility, specimens should be subjected to either Millipore filtration or immunoprecipitation prior to analysis, particularly when correlation of DNA polymerase activity with HBeAg and its corresponding antibody is attempted.

One hundred ninety-one varicella-susceptible children with leukemia in remission were immunized with live attenuated varicella vaccine. There was serological evidence of an immune response in approximately 80% after one dose and in more than 90% after two doses. The major side effect was mild to moderate rash, seen especially in children with maintenance chemotherapy suspended for one week before and one week after vaccination. Children with rash had higher antibody titers than those without rash, but those with rash were also at risk (10%) to transmit vaccine virus to others. Twenty-two vaccinees subsequently had household exposures to varicella or zoster. The attack rate of clinical varicella in these vaccinees was 18%, significantly lower than the attack rate of approximately 90% in varicella-susceptible persons with household exposures. All cases of clinical illness were extremely mild, with an average of about 50 vesicles. The mild character of the illness was clearly different than varicella in unimmunized children receiving chemotherapy for leukemia. Varicella vaccine was approximately 80% effective in preventing clinical varicella in children with leukemia and completely effective in preventing severe varicella in this high-risk group. (JAMA1984;252:355-362)

The Oka vaccine strains of varicella-zoster virus (VZV) have a significantly different BgII DNA restriction pattern from that of American wild-type isolates of VZV. This difference consists primarily of an additional BgII site, which lies within the BamHI "D" fragment. In conjunction with a study of the efficacy of an experimental Merck/Oka VZV vaccine, the area of the genome from which the most marked restriction pattern alteration arises was studied more closely to determine if there are other significant differences between the Oka strains and American wild-type strains. BamHI "D" fragments from the DNA of the Oka parent strain (the progenitor of the vaccine strain), the RIT/Oka vaccine strain (a derivative of the Oka parent strain), the Merck/Oka vaccine strain, and the EF strain (an American wild type), were submitted to extensive endonuclease digestion studies to ascertain if additional unique restriction sites are present in the Oka parent or vaccine strains. The extra BgII restriction site characteristic of the Merck/Oka vaccine strain is also present in the DNA of the parent virus as well as its derivatives and was therefore not produced by the "attenuation" process. No other novel sites were found in the Oka parent or Oka-derived strains in this section of the genome. The Merck/Oka vaccine strain of VZV, despite its Japanese origin, is therefore quite similar to circulating American varicella-zoster virus strains. Varicella-zoster virus DNA, at least in the area of the BamHI D fragment, also appears to be remarkably stable from strain to strain.

A persistent infection with varicella-zoster virus was established in the Mewo human melanoma cell line. This persistently infected cell line went through periodic crises of virus-induced cell killing and then recovery. Analyses of viral DNA derived from the persistently infected cultures revealed that novel viral nucleic acid rearrangements had been generated. These viral DNA sequences were derived from a specific region of the inverted repeat sequence of the genome flanking the short unique genome segment. The novel DNA was of various lengths, each generated by tandem duplication of an approximately 2760 base pair sub-sequence of the normal viral inverted repeat. These novel sequences were inserted into an otherwise apparently normal genome.

Infection with simian virus 40 is known to induce many cells to synthesize DNA and to divide in a medium lacking serum protein growth factor(s) that is essential for growth of uninfected cells (factor-free medium). Cells infected under these conditions then go through several rounds of division, since colonies containing more than 100 cells are formed. Many of these colonies are abortively transformed since, upon subsequent passage of the cells in standard medium, they can no longer grow in factor-free medium and show no other properties of viral transformation. We have examined these abortively transformed cells for the presence of simian virus 40 DNA sequences. Of the three clones tested, two were found to contain viral genetic material despite the fact that they were phenotypically normal. The number of simian virus 40 genome equivalents present was determined by measurement of DNA reassociation kinetics on hydroxyapatite. Two of the abortively transformed lines contained approximately five viral genome equivalents per diploid cell, while the DNA from a third abortive transformant was indistinguishable from that of uninfected BALB/3T3 cells. A standard simian virus 40 transformant, isolated under similar conditions, contained two copies of the viral genome per cell. The abortive transformants also appear to contain the entire viral genome rather than multiple partial copies. Subclones of one abortively transformed line containing five copies per cell had 2.7-10 copies of viral genetic material per diploid cell.

Kinetic analysis of the reassociation of DNA synthesized in vitro by a murine leukemia virus DNA polymerase revealed two classes of double-stranded product representative of 25 and 100 percent of viral genetic information. The DNA product representing the smaller portion of the viral genome comprised 85 percent of the double-stranded DNA generated in vitro and was extensively duplicated in the genomes of both normal cells and cells containing RNA tumor virus.

Alterations occur in the supercoiled form of viral DNA after the serial undiluted passaging of simian virus (SV) 40. We have identified a portion of the viral genome which is amplified during this process. These SV40 DNA sequences represent about 30% of the viral genetic information and are present in a reiterated form in twisted circular molecules prepared from purified virions. In addition, reiterated and unique green monkey DNA sequences are incorporated into supercoiled viral DNA. The cellular DNA appears to be inserted at numerous locations in the DNA I molecules.

The number of SV40 ‡ genome equivalents present in green monkey and SV40 transformed mammalian DNA's have been evaluated by measuring DNA reassociation kinetics on hydroxyapatite. Under the proper conditions, this method is sufficiently sensitive to detect less than one SV40 DNA molecule per mammalian genome (one part in 106) as shown by reconstruction experiments. In four out of five SV40 transformed lines examined, an average of one SV40 genome equivalent was present in the cell DNA; three SV40 DNA equivalents per cell were found in the fifth viral transformed line. The background level of SV40 DNA sequences within the 3T3 genome was 0.45 equivalent per cell. An average of 0.5 SV40 genome equivalent was measured per African green monkey genome, an amount too small to be reliably detected using DNA-DNA hybridization on nitrocellulose membranes. The biological significance of these results and their relationship to previously reported values are discussed.

To determine whether alternating regimens consisting of zidovudine and 2',3'-dideoxycytidine (ddC) reduce the toxicity and maintain or increase the antiretroviral effect associated with each drug alone.An unblinded, randomized (phase II) clinical trial in which seven treatment regimens were compared.Outpatient clinics of 12 AIDS Clinical Trials Units.One hundred thirty-one patients with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex and serum p24 antigenemia (or = 70 pg/mL).Treatments included weekly or monthly alternating zidovudine (200 mg every 4 hours) and ddC (0.01 or 0.03 mg/kg body weight every 4 hours); weekly intermittent zidovudine, 200 mg every 4 hours, or ddC, 0.03 mg/kg every 4 hours; and continuous zidovudine.Toxicity, CD4 cell counts, serum p24 antigen levels, and clinical end points. Data were analyzed for the first 48 weeks of therapy (median follow-up, 40 weeks).Hematologic toxicity was significantly less frequent in patients who received zidovudine therapy every other week (11% to 15%) or every other month (11% to 14%) than in those who received continuous zidovudine therapy (33%) (P0.02). Weekly alternating therapy with zidovudine and ddC, 0.03 mg/kg, or intermittent therapy with ddC, 0.03 mg/kg, produced high rates of peripheral neuropathy (41% and 50%, respectively). Neuropathy occurred in 10% to 21% of patients in the other three alternating-therapy limbs and in 17% of patients receiving zidovudine alone (intermittently or continuously). Initial increases in CD4 cell counts were sustained in three alternating-therapy limbs, but counts returned to baseline by week 28 in the remaining limbs. The median weight gain at week 48 was significantly greater in patients treated with alternating regimens (0.9 to 3.8 kg) compared with those treated with continuous zidovudine therapy (-0.7 kg) (P = 0.008). Patients treated with alternating regimens and those treated with continuous zidovudine had similarly sustained decreases in p24 antigen levels.These findings suggest that alternating therapy with zidovudine and ddC reduces the toxicity associated with each drug alone while maintaining strong antiretroviral activity.

Varicella may be severe in children with leukemia, with 10% mortality. We therefore evaluated the efficacy of live attenuated varicella vaccine in children with acute lymphocytic leukemia (ALL) in remission. We immunized 53 children with ALL off chemotherapy and 138 with ALL with chemotherapy suspended for 2 weeks, with 1000 pfu of varicella-zoster (VZ) (Oka) vaccine. After 1 dose of vaccine 82% seroconverted; 95% seroconverted after 2 doses, as detected by the fluorescent antibody to membrane antigen (FAMA) assay.