Reproducibility of regional placental vascularity/perfusion measurement using 3D power Doppler.

OBJECTIVES: To assess reproducibility and regional variability of placental perfusion measurement using three-dimensional (3D) power Doppler VOCAL() (Virtual Organ Computer-aided AnaLysis). METHODS: Twenty pregnant women at 26-34 weeks' gestation with normally grown, biophysically normal, singleton pregnancies with anterior placentae had placental power Doppler mapping data stored digitally from each of the four placental quadrants. Each was imaged by two investigators, with two datasets stored per investigator per quadrant. 5760 data values from the 320 datasets were evaluated by the same two investigators. Power Doppler imaging of the placental cord insertion was performed to generate a value for standardization as 'fractional moving blood volume' if appropriate. The vascularization index (VI), flow index (FI) and vascularization flow index (VFI) were calculated from spherical regions-of-interest to assess reproducibility within and between quadrants and between investigators for both acquisition and analysis. RESULTS: We found extensive variability for all readings. For repeated measurements within the same dataset the intra-analyzer intraclass correlation coefficient (ICC) range was: 0.24-0.57 for VI, 0.33-0.78 for FI and 0.12-0.48 for VFI. The corresponding interanalyzer ICC range was: 0.38-0.92 for VI, 0.40-0.85 for FI and 0.10-0.92 for VFI. The intra-acquirer variability (paired t-test) mean differences range was: - 3.91 to 4.71 for VI, - 2.68 to 3.31 for FI and - 2.23 to 2.78 for VFI; the corresponding interacquirer variability (paired t-test) range was: - 1.92 to 5.18 for VI, - 3.06 to 2.04 for FI and - 1.69 to 2.60 for VFI. The regional variability range (coefficient of variation) was: 6.28-126.34% for VI, 2.26-49.01% for FI and 6.09-151.55% for VFI. For all analyzed data, FI showed least variability and cord values for VI were consistently 100% (mean VFI, 98.4 and 98.8 between observers). CONCLUSIONS: There is insufficient evidence to support the meaning, reliability or reproducibility of VOCAL (VI, FI or VFI) as a tool to quantify placental perfusion, despite its use in multiple publications and journal submissions. There is poor reproducibility at the most fundamental level. Further investigation into the reproducibility of placental perfusion and quantification using VOCAL is required before development and application as a clinically useful tool.

In vitro evaluation of cell-mediated immunity to Epstein-Barr herpesvirus by cell migration inhibition tests.

Migration of peripheral leukocytes in samples from sensitized [Epstein-Barr virus (EBV) antibody-positive] humans was greatly inhibited when challenged by antigen prepared from EBV-producing P3HR-1 cells but not by antigen prepared from EBV-nonproducing RAJI cells, EBV-negative human fibroblasts, or epithelial cells. Such inhibition was not observed when peripheral leuocytes from subjects or neonates not sensitized to EBV were challenged. Similar results were obtained in a two-stage test when the same leukocyte samples were challenged in vitro by antigen prepared from P3HR-1 cells and the cell-free supernatant was assayed for migration inhibition factor (MIF) in the guinea pig macrophage migration inhibition test; migration of guinea pig peritoneal exudate cells was greatly inhibited by the supernatant filtrates of leukocyte cultures only from subjects positive for EBV-antibody. Furthermore, this inhibitory effect was not observed if supernatant filtrates from leukocyte cultures challenged by antigens prepared from RAJI cells, fibroblasts, or epithelial cells were used. The EBV antigen transformed peripheral leukocytes and induced early antigen production in RAJI cells; however, a "killed" preparation (by UV irradiation) was sufficient for eliciting MIF production.

Granulocytic colonies on macrophage-coated cellulose acetate membranes (CAM) in S1/S1dmice.

The kinetics of formation of granulocytic colonies on macrophage-coated cellulose acetate membranes (CAM) were investigated in Sl/Sld mice, which have a genetic defect in their hematopoietic microenvironment. CAM were left in the peritoneal cavity of mice for 7 days to become coated with peritoneal cells. The mice were then sublethally irradiated to suppress endogenously derived granulocytic colonies, and bone marrow cells were injected i.p. within 1--2 hr irradiation. During the next 7 days, peroxidase-positive granulocytic colonies appeared on CAM. The number of colonies on CAM in Sl/Sld mice at 7 days was consistently less than that from littermate +/+ mice. Since it appeared that the CAM in the Sl/Sld provided an inferior microenvironment to that of the +/+, the effect of raising a CAM in one genotype and transferring it to another was investigated. CAM raised in +/+ or in Sl/Sld were transferred to an irradiated +/+ or Sl/Sld following which cells were injected into the secondary host and colonies determined subsequently. The number of granulocytic colonies on CAM was influenced primarily by the type of secondary host. Colony number was consistently less in the secondary Sl/Sld host than in the +/+ host whether the primary host was a Sl/Sld or a +/+. These observations confirm the concept of a microenvironmental defect in Sl/Sld mice. In addition, the studies indicate that the microenvironment on a CAM can be modified by a secondary host and suggest that "remodeling" of CAM may be a continuous, kinetic process.

Antimicrobial and chemopreventive properties of herbs and spices.

Herbs and spices have been used for generations by humans as food and to treat ailments. Scientific evidence is accumulating that many of these herbs and spices do have medicinal properties that alleviate symptoms or prevent disease. A growing body of research has demonstrated that the commonly used herbs and spices such as garlic, black cumin, cloves, cinnamon, thyme, allspices, bay leaves, mustard, and rosemary, possess antimicrobial properties that, in some cases, can be used therapeutically. Other spices, such as saffron, a food colorant; turmeric, a yellow colored spice; tea, either green or black, and flaxseed do contain potent phytochemicals, including carotenoids, curcumins, catechins, lignan respectively, which provide significant protection against cancer. This review discusses recent data on the antimicrobial and chemopreventive activities of some herbs and spices and their ingredients.

Antigen competition in cytotoxic T cell response to minor histocompatibility antigens.

Two populations of class-I-restricted CTL are generated from BDF1 mice against multiple minor-HA when BALB.B cells carrying multiple minor-HA are used as immunogen, whereas one population of Db-restricted CTL is generated when H-25.3 antigen on B6.C-H-25c cells is used as antigen. CTL generated against H-25.3 antigen on B6.C-H-25c cells are cytolytic to B6.C-H-25c and BALB.B targets, whereas CTL generated in the same manner with multiple minor-HA on BALB.B cells kill BALB.B targets only. This must be an instance of antigen competition, because H-25.3 antigen is immunostimulatory when used on its own, but not when accompanied by other minor-HA. This competition effect is not an artefact seen during the generation of CTL in MLC cultures in vitro, because it is also seen during the generation of CTL in vivo. We have not identified the immunodominant antigen(s) on BALB.B cells. We have studied this form of antigen competition in some detail. First, BDF1 spleen primed in vivo to multiple minor-HA on BALB.B cells do not respond in vitro to restimulation by H-25.3 antigen, suggesting that antigen competition is not mediated by suppressor cells, but that BALB.B cells do not prime BDF1 spleen cells against H-25.3 antigen in vivo. Second, BDF1 spleen cells primed to the same multiple minor-HA carried on F1(BALB/c X B6.C-H-25c) cells respond to restimulation by H-25.3 antigen in vitro. Third, BDF1 spleen cells primed in vivo to H-25.3 antigen on B6.C-H-25c cells do not respond in vitro to restimulation by multiple minor-HA on BALB.B cells, but they do respond to F1(BALB/c X B6.C-H-25c) cells that carry the same multiple minor-HA as BALB.B cells, and generate Db-restricted anti-H-25.3 CTL. Fourth, BDF1 spleen cells primed in vivo and boosted in vitro with B6.C-H-25c cells readily develop CTL to H-25.3 antigen, whereas when the BDF1 mice are exposed to H-25.3 antigen in vivo by repeated footpad injections of B6.C-H-25c cells, CTL to H-25.3 antigen do not develop. However, anti-H-25.3 CTL do develop after H-25.3 antigen-bearing B6.C-H-25c cells and antigen-specific Th cells are injected into preprimed BDF1 mice. These results are discussed with respect to possible mechanisms of antigen competition.

Modification of human immunodeficiency viral replication by pine cone extracts.

We have shown previously that two fractions (PC6 and PC7) extracted from cones of the Japanese white pine Pinus parvifloria Sieb. et Zucc have potent immunopotentiating effects. Here, we show that PC6 and PC7 inhibited HIV-1 replication (greater than 95%), in a dose-dependent manner, in chronically infected CR10/HIV-1 cells and in acute cytolytic HIV-1 infection of CEM cells. Treatment of CEM cells, prior to or after acute infection with HIV-1, reduced subsequent viral production, but the best inhibitory effect was obtained with treatment before and after infection: an 80% inhibition was achieved with as little as 3 micrograms/ml of PC6. Comparable results were also obtained when PC6 was used to inhibit HIV-1 replication in the U937 human histiocytic lymphoma cell line. Both PC6 and PC7 were relatively nontoxic to cells. The anti-HIV-1 effect of PC6 and PC7 we observed in this report, coupled with earlier reports of their immunopotentiating properties suggest their potential as ideal therapeutic agents for the treatment of AIDS.

Inhibition of human immunodeficiency virus forward and reverse transcription by PC6, a natural product from cones of pine trees.

We have previously shown that PC6, a natural product extracted from cones of Pinus parviflora Sieb et Zucc, can inhibit the replication of human immunodeficiency virus type 1 (HIV-1) in CD4+ T cells and in monocyte/macrophage cell lines. Here, we show by immunoprecipitation of HIV-1 proteins with a specific pooled serum that PC6 inhibited the expression of all HIV-1 proteins in CEM cells. PC6 did not affect the posttranslational processing of the HIV-1 proteins. Northern, Southern, and kinetics analyses revealed that PC6 inhibited HIV-1 reverse and forward transcription in CEM cells. Transient transfection of CEM cells with HIV-1 long terminal repeat (LTR) linked CAT DNA also showed that PC6 inhibited LTR-driven gene expression at the transcriptional level.

Superinfection of a defective human immunodeficiency virus type 1 provirus-carrying T cell clone with vif or vpu mutants gives cytopathic virus particles by homologous recombination.

The partially CD4-expressing T cell clone, Vpr-1, which carries a latent vpr-defective HIV-1 genome and expresses HIV-1 Nef protein only, was permissive to superinfection by HIV-1. Superinfection of Vpr-1 with vif- or vpu-defective mutants, which were noncytopathic, reactivated the vpr-defective virus and led to homologous recombination and cytopathogenesis. The data provide an experimental model for homologous recombination being an important mechanism whereby HIV-1 acquires genetic heterogeneity, and when occurring among defective virus in vivo bestows novel biological activities and virulence.

Soluble interleukin 2 receptors in patients with malignant lymphoma.

We performed a retrospective analysis of serum interleukin 2 receptor (IL-2R) levels in a group of 35 patients with malignant lymphoma (ML; 13 T cell and 22 B cell) using a new enzyme immunoassay. Our objectives were to determine if elevated levels of soluble IL-2R occur in patients with active ML, whether serum IL-2R levels are prognostic, and whether prospective studies are warranted. Our preliminary data indicate that serum IL-2R levels correlate with disease activity and size of tumor, but not with grade or stage of the tumor. Five-year actuarial survival was 20% for patients with IL-2R levels greater than 1000 U/mL at any time during their course and 86% for patients who did not exceed that threshold. Furthermore, patients with IL-2R levels lower than 1000 U/mL were more likely to achieve a complete remission. Serum lactate dehydrogenase and uric acid levels did not show significant correlation with disease activity or prognosis. We conclude that serum IL-2R levels may have clinical and prognostic significance in patients with ML and that prospective studies are indicated.

Prevalence of circulating antibodies to p10, a non-structural protein of the Borna disease virus in cats with ataxia.

Japanese domestic cats were surveyed for circulating antibodies to the plO and p24 proteins of the Borna disease virus (BDV) by Western blotting. Twenty-four of 52 cats (46.2%) with ataxia and other neurologic symptoms of unknown cause were positive for antibodies to BDV p10 and/or p24. In contrast, cats without neurological symptoms gave a significantly lower prevalence of anti-BDV antibodies to p10 and/or p24 (36 of 152 cats, 23.7%). Thirty specific pathogen-free (SPF) cats tested as controls were uniformly negative to BDV pl0 and p24 antigens. These results suggest that BDV may play a role in ataxia in cats. Additionally, our results suggest that it is necessary to use both p10 and p24 as antigens to detect circulating antibodies to BDV in cats.

Infectious mononucleosis: recognition and management.

Differentiation from other common viral infections depends on a triad of clinical, hematologic, and serologic determinations. A distinctive feature is that while a number of circulating immunoglobulins are often detected, only Paul-Bunnell antibodies, found in 75% of cases, are specific for IM. Symptomatic care only is indicated for most patients, though severe cases or complications may require antibiotics or corticosteroids.

Helper T cell lines to major and to minor histocompatibility antigens are equally effective in the regulation of B cell responses in vivo.

Long-term lines of helper T (Th) cells, reactive to minor histocompatibility (minor-H) antigens, were grown by antigen restimulation in the absence of exogenous interleukin-2. These lines were antigen specific and H-2b restricted. When introduced in vivo by adoptive transfer, these Th cells helped syngeneic B cells in an antibody response to other alloantigens. Linked recognition was required for effective help to occur, this suggests B cell presentation of antigen to Th cells in vivo. Parallel titration experiments performed with long-term cultured Th lines to MHC and to minor-H antigens showed that, on a per cell basis, they are equivalent in their ability to help in vivo B cell responses. This shows that any inability to produce antisera to minor-H antigens is not due to a Th or APC defect, but results from either a B cell defect or from suppression.

T helper cell lines that augment in vivo cytotoxic T-cell responses to minor alloantigens.

We have investigated the ability of long-term cultured T helper (Th) cell lines to help an in vivo cytotoxic T lymphocyte (CTL) response to non-H-2 alloantigens (minor antigens). Th cell lines specific for various single or undefined minor antigens were selected by regular restimulation with antigen in vitro. They were antigen specific and H-2 restricted in proliferation assays and were found to be able to help primary CTL responses to multiple minor antigens and secondary CTL responses to single minor antigens. Although the Th were antigen specific they did not determine the specificity of the CTL. Th cells were both necessary and limiting for an effective CTL response indicating that "helper-independent" CTL are not in themselves sufficient to generate a strong in vivo response. Under conditions where a CTL response was clearly H-2 restricted, Th cells were not. Thus, the Th cells appeared to be activated by reprocessed antigen rather than antigen on the surface of the injected antigenic cells even though the CTL themselves reacted directly to the injected antigen.