Cold stress-induced neuroinvasiveness of attenuated arboviruses is not solely mediated by corticosterone

In previous studies we have shown that various stress paradigms can induce the penetration of noninvasive, attenuated viruses into the central nervous system (CNS). Since glucocorticoids levels are elevated during stress, we compared the effect of cold stress and corticosterone (CS) injection on neuroinvasiveness of a non-invasive encephalitic virus, WN-25 (West Nile). Exposure of inoculated mice to cold stress or CS resulted in high viremia and a marked increase in mortality when compared to control untreated mice. Exposure of WN-25 inoculated mice to cold treatment or CS injection led to high blood virus levels as compared to nontreated mice (3.2 and 3.1 vs > 1 log 10 PFU/ml). Cold stress or CS (5000 ng/mouse) treatment caused a mortality rate of 70% and 50% of the WN-25 inoculated mice respectively. No mortality was recorded in control inoculated groups (p < 0.05). Passive transfer serum from uninfected cold stressed mice to WN-25 inoculated nonstressed mice, resulted in similar mortality. The levels of CS in passive transferred serum from cold stressed animals was 500 ng/ml, only 2% (100 vs. 5000 ng) of the CS dose required to obtain a similar effect on viral penetration and mortality when CS was injected directly. Therefore, we concluded that CS was not the sole factor responsible for the cold stress effect on the viral infection outcome.     

Primary human immunodeficiency virus type 1 viremia and central nervous system invasion in a novel hu-PBL-immunodeficient mouse strain

We established four new mouse strains with defective T and B cells as well as defects in innate immunological reactions using an NK cell depletion antibody and showed that all mutant mouse strains efficiently received human peripheral blood leukocyte (PBL) engraftment (hu-PBL-scid mice). Higher levels of human immunodeficiency virus type 1 (HIV-1) replication were observed in these new hu-PBL-scid mice than in conventional hu-PBL-C.B-17-scid mice. In one particular strain, hu-PBL-NOD-scid mice, high levels of HIV-1 viremia (more than 10(6) 50% infectious doses per ml) were detected after infection with HIV-1. The plasma viral load was about 100 to 1,000 times higher than that observed in other hu-PBL-scid mice infected with HIV-1. Although high-level viremia did not correlate with the total amount of HIV-1 RNA in cells from infected mice, high levels of free virions were detected only in hu-PBL-NOD-scid mice. HIV-1 viremia induced systemic HIV-1 infection involving the liver, lungs, and brain. PCR in situ hybridization confirmed that HIV-1-infected cells invaded the brain tissue of the hu-PBL-NOD-scid mice. Our results suggest that the genetic background, including innate immunity, is critical in the development of primary HIV-1 viremia and subsequent central nervous system invasion with HIV-1. The hu-PBL-NOD-scid mouse represents a useful model for the study of the pathogenesis of HIV-1 in vivo, especially brain involvement, and therapy of primary HIV-1 viremia.     

Use of dental instruments for bridging during electric pulp testing

It has been suggested that dental instruments can be used as bridging instruments to facilitate electric pulp testing of teeth with extensive restorations. This study reports a clinical investigation to evaluate the effectiveness of this procedure. One hundred seventeen vital teeth in 20 volunteers were tested. Ten endodontically treated teeth functioned as controls. Following appropriate isolation and asepsis technique, baseline recordings of the threshold response with the electric pulp tester were taken. With the use of dental explorers and endodontic files to bridge between the probe tip and the tooth surface, recordings were made of the threshold responses. Findings indicate that electrically conductive dental instruments can be reliably used as bridging instruments with the electric pulp tester.     

Deletion of nef slows but does not prevent CD4-positive T-cell depletion in human immunodeficiency virus type 1-infected human-PBL-SCID mice

The pathogenicity of four human immunodeficiency virus type 1 (HIV-1) isolates with nef deleted for SCID mice repopulated with human peripheral blood leukocytes (hu-PBL-SCID mice) was studied. Deletion of nef led to a substantial reduction in CD4-positive T-cell depletion and delayed kinetics of plasma viremia in infected hu-PBL-SCID mice. Deletion of the nef gene impacts both the efficiency of primary infection and the cytopathicity of virus for infected CD4-positive T cells in this animal model of HIV-1 infection.     

Effect of foscarnet on quantities of cytomegalovirus and human immunodeficiency virus in blood of persons with AIDS

Four intravenous dosages of foscarnet given for 10 days were compared with no therapy in persons with AIDS who had asymptomatic cytomegalovirus (CMV) viremia. CMV viremia was quantitated by endpoint cell dilution microcultures, pp65 antigenemia assay, and measurement of CMV DNA in peripheral blood leukocytes by a quantitative-competitive PCR. Human immunodeficiency virus type 1 (HIV-1) viremia was quantitated by endpoint cell dilution microculture, serum p24 antigen assay, and PCR for HIV-1 RNA in plasma. Twenty-seven subjects who had received a median of 22 months of nucleoside antiretroviral therapy were enrolled. Twenty-two subjects received foscarnet, which was well tolerated and decreased the CMV burden, as reflected by all three indicator assays. During the 10 days of dosing, the level of CMV viremia, as measured by 50 percent tissue culture infective doses, decreased from 117.5 to 12.7 (P = 0.001), the amount of CMV DNA decreased from 20,328 copies to 622 copies per 150,000 leukocytes (P = 0.02), and the level of CMV pp65 antigenemia decreased from 14.9 to 1.6 positive peripheral blood mononuclear cells per 50,000 leukocytes (P = 0.008). A significant pharmacodynamic relationship was found between the peak foscarnet concentration and a decrease in the level of CMV antigenemia (P < 0.05). Foscarnet had no effect on quantitative HIV-1 microcultures during the 10 days of treatment, but the HIV-1 p24 antigen level in serum decreased significantly, from 454 to 305 pg/ml (P = 0.01). Also, a significant pharmacodynamic relationship was seen between plasma HIV-1 RNA concentrations and both peak foscarnet concentration (P < 0.01) and the area under the foscarnet time-concentration curve (P < 0.05). Reductions in the levels of CMV and HIV-1 viremia correlated quantitatively with systemic exposure to foscarnet, whereas control subjects actually experienced an increase in CMV and HIV-1 burdens. The dual antiviral activity of foscarnet shown in this trial encourages investigation of its use in combination with other antiretroviral therapies for persons with AIDS.     

Importance of localized skin infection in tick-borne encephalitis virus transmission

Arboviruses are transmitted to vertebrates by the "bite" of infected arthropods. Events at the site of virus deposition are largely unknown despite increasing evidence that blood-sucking arthropods immunomodulate their skin site of feeding. This question is particularly relevant for ixodid ticks that feed for several days. To examine events under conditions mimicking tick-borne encephalitis (TBE) virus transmission in nature (i.e., infected and uninfected Ixodes ricinus ticks feeding on the same animal), infected adult and uninfected nymphal ticks were placed in one retaining chamber (skin site A) and uninfected nymphs were placed within a second chamber posteriorly (skin site B) on two natural host species, yellow-necked field mice (Apodemus flavicollis) and bank voles (Clethrionomys glareolus). Virus transmission from infected to uninfected cofeeding ticks was correlated with infection in the skin site of tick feeding. Furthermore, virus was recruited preferentially to the site in which ticks were feeding compared with uninfested skin sites. Viremia did not correspond with a generalized infection of the skin; virus was not detected in an uninfested skin site (C) of 12/13 natural hosts that had viremia levels > or = 2.0 log10 ic mouse LD50/0.02 ml blood. To characterize infected cells, laboratory mouse strains were infested with infected ticks and then explants were removed from selected skin sites and floated on culture medium. Numerous leukocytes were found to migrate from the skin explants of tick feeding sites. Two-color immunocytochemistry revealed viral antigen in both migratory Langerhans cells and neutrophils; in addition, the migratory monocyte/macrophages were shown to produce infectious virus. The results indicate that the local skin site of tick feeding is an important focus of viral replication early after TBE virus transmission by ticks. Cellular infiltration of tick feeding sites, and the migration of cells from such sites, may provide a vehicle for transmission between infected and uninfected cofeeding ticks that is independent of a patent viremia. The data support the hypothesis that viremia is a product, rather than a prerequisite, of tick-borne virus transmission.     

Cell-associated viral RNA expression during acute infection with HIV type 1

The mechanism of decline in viremia following acute infection with HIV is unknown. To characterize this process virologically, the expression of viral RNAs was analyzed in samples of peripheral blood mononuclear cells (PBMCs) from a patient who experienced a 100-fold decline in plasma viremia over a 13-day period prior to the initiation of antiretroviral therapy. Cell-associated viral RNA declined in association with the decline in plasma virus. During the initial 7 days of observation, plasma viremia declined more than 10-fold with no change in the ratio of unspliced to multiply spliced mRNAs. The efficiency of viral gene expression did not decline during the study period and varied from 380 to 2800 unspliced RNA copies per productively infected cell. Together, these data indicate no change in the relative proportion of cells in late- and early-stage gene expression during the initial decline and provide evidence against shortening of the viral replication cycle by immune surveillance. However, the prevalence of productively infected cells declined markedly during the 13 days of observation, from 1 in 250 to 1 in 25,000 PBMCs. These data are compatible with depletion of available target cells during the initial decline in viremia. As the level of plasma virus stabilized after 8 days of observation, the ratio of unspliced to multiply spliced mRNAs rose; this rise was due to a relatively greater decline in multiply spliced mRNA. These data suggest the possible onset of a blockade to new infection events (for example, by neutralizing antibody or chemokines), causing an increase in the relative proportion of cells in late-stage gene expression. They may also be explained, however, by the persistence of cell-associated virions together with the near disappearance of productively infected cells from the circulation.     

Distribution of HIV type 1 (HIV-1) in blood components: detection and significance of high levels of HIV-1 associated with platelets

BACKGROUND: Although inactivation of enveloped viruses transmitted by plasma derivatives has been successful, no methods for virus inactivation or removal have been established for platelet concentrates or red cell (RBC) components. Relatively little is known regarding the extent or significance of virus interactions with the cellular constituents in these components. STUDY DESIGN AND METHODS: Units of whole blood were collected from six HIV type 1 (HIV-1)-positive, asymptomatic individuals and separated into peripheral blood mononuclear cells (PBMNCs), cell-free plasma, white cell-reduced platelet concentrate, and white cell-reduced RBCs. DNA and RNA polymerase chain reaction and virus culture methods were used to study the compartmentalization of HIV-1 immediately after component preparation and after storage. RESULTS: As expected, HIV DNA and infectious virus were detected in fresh blood and in PBMNCs, and virion-associated RNA was detected in fresh plasma from all six donors. The levels of viral nucleic acids in these preparations remained relatively stable with 4 degrees C storage, whereas infectivity of PBMNCs was rapidly lost. Washed RBCs tested negative for HIV in all assays at all time points. Platelets retained high levels of HIV RNA (but not infectivity) after extensive washing, as well as after storage at 4 and 22 degrees C. High-level platelet-associated HIV-1 was also demonstrated in samples collected during early seroconversion. Periseroconversion and postseroconversion levels of platelet-associated HIV-1 correlated with the level of plasma viremia and with the rate of progression to AIDS. Cell-free virus from donor plasma and tissue culture fluid rapidly and firmly attached to platelets from noninfected donors. Infectivity of tissue culture virus bound to platelets was demonstrated in vitro. CONCLUSION: Significant levels of HIV-1 are associated with platelets during all stages of infection. Platelet-associated HIV could either mediate virus clearance or facilitate virus dissemination and expanded tropism. Finally, virus inactivation research must address virus associations with platelets.     

Trends in glove use by dentists in England and Wales: 1989-1992

The routine wearing of gloves during the dental treatment of patients forms an integral part of the recommended guidelines of many authorities worldwide. Compliance has been variable but there is evidence to show that this is increasing, as are other cross-infection control measures. A survey of glove use by general dental practitioners in England and Wales was carried out in 1989. This paper reports a follow up study in 1991/2 which aimed to identify any differences that had occurred in the interim. The results indicated that there was increasing compliance with recommended cross-infection control measures, although only one fifth of respondents were found to change gloves between patients. Of those practitioners who did not change gloves between patients, half considered that blood contact was not a reason for changing gloves. Comparisons are made with compliance rates in other countries and recommendations made for further education within the dental profession on the subject of cross-infection control.     

Role of Anopheles quadrimaculatus and Coquillettidia perturbans (Diptera: Culicidae) in the transmission cycle of Cache Valley virus (Bunyaviridae: Bunyavirus) in the midwest, USA

Midwestern populations of Coquillettidia perturbans (Walker) and Anopheles quadrimaculatus (Say) were tested for their ability to transmit Cache Valley virus (CV), a recognized human and animal pathogen. Field-collected mosquitoes were fed artificial blood meals containing 5.2-6.2 log10 pfu/ml of CV. After 9-23 d at 28 degrees C, 75-93% of blood-fed Cq. perturbans had disseminated infections and 6-62% transmitted the virus to suckling mice. However, when infected with a lower virus titer (3.3 log10 pfu/ml), only 10-36% of the mosquitoes had disseminated infections and 0-10% transmitted the virus to suckling mice. A similar infection rate (21%) was observed in Cq. perturbans fed on viremic (3.2 log10 pfu/ml) hamsters. An. quadrimaculatus were infected (81-100%) by both doses used, with transmission rates ranging from 13-67% after 16-23 d of incubation. Transmission rates for the laboratory strain An. quadrimaculatus SAVANNAH ranged from 20 to 33% after 7-14 d of incubation. Our data show that although An. quadrimaculatus is more susceptible to CV infections than Cq. perturbans, both mosquito species could be involved in the midwestern transmission cycle of the virus.     

Accumulation of human immunodeficiency virus-specific cytotoxic T lymphocytes away from the predominant site of virus replication during primary infection

Down-regulation of the initial burst of viremia during primary human immunodeficiency virus (HIV) infection is thought to be mediated predominantly by HIV-specific CD8+ cytotoxic T lymphocytes (CTL). This response is associated with major perturbations in the T cell receptor (TCR) repertoire. To investigate the failure of the cellular immune response to adequately control viral spread and replication and to prevent establishment of HIV infection, changes in the TCR repertoire and in the distribution of virus-specific CTL between blood and lymph node were analyzed in three patients with primary infection. By the combined use of clonotype-specific polymerase chain reaction and analysis of the frequency of in vivo activated HIV-specific CTL, it was shown that HIV-specific CTL clones preferentially accumulated in blood as opposed to lymph node. Accumulation of HIV-specific CTL in blood occurred prior to effective down-regulation of virus replication in both blood and lymph node. These findings should provide new insights into how HIV, and possibly other viruses, elude the immune response of the host during primary infection.     

Treatment with enalapril modifies the pain perception pattern in hypertensive patients

The cardiovascular system shares numerous anatomic and functional pathways with the antinociceptive network. The aim of this study was to investigate whether angiotensin-converting enzyme (ACE) inhibitor treatment could affect hypertension-related hypalgesia. Twenty-five untreated hypertensive patients, together with a control group of 14 normotensive subjects, underwent dental pain perception evaluation by means of a pulpar test (graded increase of test current applied to healthy teeth). After the evaluation of the dental pain threshold (occurrence of pulp sensation) and tolerance (time when the subjects asked for the test to be stopped), all the subjects underwent a 24-hour ambulatory blood pressure monitoring. The hypertensive group then was treated with 20 mg/d enalapril, whereas the normotensive subjects remained without any treatment. After a time interval of 6+/-2 months, the dental pain sensitivity was retested in all the subjects, and ambulatory blood pressure was recorded during treatment in the hypertensive patients. At the first assessment, hypertensive patients showed a higher pain threshold than normotensive subjects (P<.001). On retesting of pain sensitivity in hypertensive patients, a significant decrease of both pain threshold and tolerance, leading to their normalization, was observed during treatment (P<.001 and P<.005, respectively), in the presence of reduced 24-hour and office blood pressure values. A slight, though significant, correlation was observed between variations in pain tolerance and baseline blood pressure changes occurring during treatment. During follow-up, the normotensive subjects did not show any significant pain perception or office blood pressure changes. Hypertension-related hypalgesia was confirmed. Mechanisms acting both through lowering of blood pressure and specific pharmacodynamic properties may account for the normalization of pain sensitivity observed in hypertensive patients during treatment with ACE inhibitors.     

Infection control in the dental practice with emphasis on the orthodontic practice

Infections present a significant hazard in the orthodontic office because they can be transmitted by blood or saliva through direct or indirect contact, droplets, aerosols, or contaminated instruments and equipment. Because the incidence of certain microbial cross-infections in the dental environment has not been well documented, orthodontic personnel may not take the problem of cross-infection as seriously as they should, and they may transmit or contract more infections than they realize. The use of effective infection-control procedures in the orthodontic office and laboratory will prevent cross-contamination that may extend to the orthodontist, office staff, assistants, and patients. The goal of this article is to present infection control in a simple, yet comprehensive, manner, and to encourage all orthodontic practitioners to implement essential infection-control procedures in their practices.     

Characterization of a live-attenuated retroviral vaccine demonstrates protection via immune mechanisms

Live-attenuated retroviruses have been shown to be effective retroviral vaccines, but currently little is known regarding the mechanisms of protection. In the present studies, we used Friend virus as a model to analyze characteristics of a live-attenuated vaccine in protection against virus-induced disease. Highly susceptible mice were immunized with nonpathogenic Friend murine leukemia helper virus (F-MuLV), which replicates poorly in adult mice. Further attenuation of the vaccine virus was achieved by crossing the Fv-1 genetic resistance barrier. The minimum dose of vaccine virus required to protect 100% of the mice against challenge with pathogenic Friend virus complex was determined to be 10(3) focus-forming units of attenuated virus. Live vaccine virus was necessary for induction of immunity, since inactivated F-MuLV did not induce protection. To determine whether immune cells mediated protection, spleen cells from vaccinated donor mice were adoptively transferred into syngeneic recipients. The results indicated that immune mechanisms rather than viral interference mediated protection.     

Obstructive sleep apnea syndrome in the Community of Valencia: current situation, study of needs and future prospects]

Experimental simian varicella virus (SVV) infection of St. Kitts vervet monkeys was evaluated as an animal model to investigate human varicella-zoster virus (VZV) infections. During the incubation period, viremia disseminated infectious virus throughout the body via infected peripheral blood lymphocytes (PBLs). A vesicular skin rash in the inguinal area, and on the abdomen, extremities, and face appeared on day 7-10 postinfection. Necrosis and hemorrhage in lung and liver tissues from acutely infected monkeys were evident upon histologic analysis. Recovery from simian varicella was accompanied by a rise in the serum neutralizing antibody response to the virus. SVV latency was established in trigeminal ganglia of monkeys which resolved the acute infection. This study indicates that experimental SVV infection of St. Kitts vervets is a useful animal model to investigate SVV and VZV pathogenesis and to evaluate potential antiviral agents and vaccines.     

Viremia in young herons and ibis infected with Venezuelan encephalitis virus

Fifty-seven of 61 nestling, 8- to 30-day-old herons of three species (Black-crowned Night Heron, Great Egret, and Snowy Egret), developed viremia lasting one to three days following subcutaneous inoculation with small doses of endemic or epidemic strains of Venezuelan encephalitis virus from Mexico, Guatemala or Venezuela. Two epidemic strains from Guatemala or Venezuela stimulated levels of viremia similar to those following infection with enzootic strains. Great Egrets, Striated and Boat-billed Herons and Scarlet Ibis older than 30 days of age developed viremias of lower levels and shorter durtions than did young birds. Marked differences in levles of viremia were not observed among Black-crowned Night Herons, Great Egrets, or Snowy Egrets. Over 50% of viremic blood samples from herons 8-30 days of age contained 1000 or more chick embryo cell culture plaque forming units of Venezuelan encephalitis per ml, levels sufficient to infect some vector species mosquitoes.