Bis-basic-substituted polycyclic aromatic compounds. A new class of antiviral agents. 7. Bisalkamine esters of 9-oxoxanthene-2,7-dicarboxylic acid, 3,6-bis-basic ethers of xanthen-9-one, and 2,7-bis(aminoacyl)xanthen-9-ones-xanthenes, and -thioxanthenes.

3,6-Bis[2-(dimethylamino)ethoxy]-9H-xanthen-9-one dihydrochloride (4, RMI 10874DA) and 1,1'-(9H-xanthene 2,7-diyl)bis[2-(dimethylamino)ethanone] dihydrochloride (16, RMI 11513DA) were found to prolong survival of mice infected with lethal challenges of encephalomyocarditis (EMC) virus. They were effective by oral as well as subcutaneous administration and showed broad-spectrum antiviral activity. They were selected for preclinical evaluation from the five series of compounds named in the title that were synthesized in analogy to tilorone and related fluorenone derivatives, described earlier. In addition to 4 and 16, compounds 11, 12, 17, and 18 showed high antiviral activity on oral as well as subcutaneous administration. High antiviral activity on subcutaneous admistration was found in the bisalkamine esters 1,2, and 14, the bis(aminoacyl)xanthenes 23 and 26, the bis(aminoalkylene)xanthene 31, the bis(aminoacyl)thioxanthenes 34-40, and the bis-basic ethers of 9-benzylide-nexanthenes 41 and 42. Structure-activity relationships showed a decrease of oral activity with increased length of side chains and increased molecular weight of dialkylamino substituents of 3,6-bis-basic ethers of xanthen-9-one and of 2,7-bis(aminoacyl)xanthenes and-xanthen-9-ones. At least one carbonyl or alkenyl function in conjugation to the xanthene nucleus either at the 9 position of the nucleus or in the side chains is required for high antiviral activity.

Genetic epidemiologic study of hearing loss in an adult population.

Previous population studies of hearing loss have been limited to children with moderate to profound impairment, and have reported that heritability accounts for at least 50% of congenital or early-onset cases. The present study was designed to assess genetic factors associated with late-onset hearing impairment in an adult population. A brief family history and audiologic questionnaire was sent to approximately 11,200 members of the consumer organization, Self Help for the Hard of Hearing, Inc., and 4,039 questionnaires were returned. All respondents reported having at least one previous audiologic exam. Reported data were verified against audiograms when available. Regardless of the reported causes, 49% of early-onset cases (< or = 20 years of age) had one or two parent(s) with some form of hearing loss compared with 62% in later-onset cases. As expected, mean age at onset was substantially younger for cases with positive family histories than cases with negative family histories. Results from nuclear segregation analysis showed that fully recessive and dominant models failed to explain the early- or late-onset hearing loss data. In this nationwide survey, the large proportion of cases with positive family histories clearly indicates the importance of genetic factors in adult-onset forms of hearing loss. Comparison with younger-onset cases will permit further delineation of differences in inheritance patterns. This study should identify more homogeneous groups of adult-onset families for further genetic study, and provide empiric information for use in genetic counselling.

Specifics on the refinement and application of two serological assays for the detection of antibodies to HHV-8.

BACKGROUND: Serologic assays for the detection of antibodies to human herpesvirus type 8 (HHV-8) are important for epidemiological studies and to further investigate the proposed pathogenesis of the virus in cancer. Although a variety of assays are available, a lack of optimization and standardization makes their usefulness uncertain, and may be responsible for the controversy concerning the prevalence of infection. OBJECTIVES: To refine an indirect immunofluorescent assay (IFA) for the detection of latent antibodies and a recombinant ORF 65 ELISA for the detection of lytic antibodies in order to increase their ability to differentiate individuals at higher and lower risk for HHV-8 infection. STUDY DESIGN: Sera from Kaposi's sarcoma (KS) patients and blood donors (BDs) were used to modify assay parameters in an attempt to better discriminate between the two populations. Modifications included methods of substrate fixation, incubation times, sample dilution, and antigen/conjugate concentrations. RESULTS: Optimal modifications to the latent IFA included acetone fixation of substrate, and dilution of sera to 1:64 which enhanced detection of HHV-8 antibodies from 68 to 92% in the KS population. Similarly, successful refinement of the ORF 65 ELISA to increase the signal-to-noise ratio included the use of 88 ng of ORF 65 antigen per well and serum dilutions of 1:50. Optical density-to-cut-off ratios directly correlated with titers, thereby introducing a strategy to predict antibody concentrations. The ORF 65 ELISA and the latent IFA were both able to discriminate between the two populations but with different efficiencies. CONCLUSIONS: Although neither the latent IFA nor the ORF 65 ELISA produced perfect test indices, improvement in their performances was noted following the optimization strategies. The ELISA produced better detection of antibodies to the virus than the IFA and permitted prediction of sample titers, thus improving cost and time effectiveness.

Prolactin and growth hormone responses in the amenorrhea-galactorrhea syndrome.

Fifteen patients with hyperprolactinemia, amenorrhea, and galactorrhea were studied; 10 of these patients had apparent tumors. Nine of 12 patients had a suppression of prolactin with oral L-dopa, and all of 4 patients undergoing dopamine infusion had suppression of prolactin. Eleven and 8 patients underwent chlorpromazine stimulation and insulin-induced hypoglycemia, respectively; none responded. Three of the 4 patients did not show a growth hormone response with dopamine infusion, and L-dopa testing failed to achieve a growth hormone response in 7 of 10 patients. The data suggest an intact dopaminergic inhibition of prolactin with exogenous agents. However, inhibition of endogenous dopamine by chlorpromazine failed to elicit a prolactin response. The disordered growth hormone response to dopaminergic agents suggests a central disorder of dopamine generation. The possible implication of these results with respect to the pathogenesis of hyperprolactinemia is discussed.

Evaluating adherence to medication in children and adolescents with HIV.

OBJECTIVE: The study was aimed to evaluate the relationship between pharmacy supply, self-reported treatment adherence and HIV viral load in HIV-infected children. METHODS: A retrospective (52 weeks) cohort study was conducted through the review of the existing databases. Pharmacy supply was classified as "home delivery" when the medications were delivered home and as "in pharmacy pick-up" when they were picked up at the pharmacy. Adherence was assessed through retrospective (3 days recall) self-report. Fisher's exact model, univariate and multivariate logistic regression analyses were used. SETTINGS: The study collected data on 140 HIV-infected children (<18 years). Adherence, pharmacy supply information and HIV viral loads were obtained from clinical and research databases. PATIENTS: The data from 127 HIV-infected children (60 boys and 67 girls; mean age 9.9 years) were collected. MAIN OUTCOME MEASURES: Complete adherence (100%) was reported in only 24% of patients. With 40% of patients being rarely or never completely adherent, 64% of children achieved undetectable viral loads during the study period. RESULTS: No association between pharmacy supply and self-reported adherence was found (p = 0.605). Self-reported adherence (p = 0.0328) and age (p = 0.025) were the significant predictors of reaching undetectable viral loads. Adolescents (>13 years) were significantly less likely to reach undetectable viral loads than children under 13 years (odds ratio 0.38; 95% CI 0.16 to 0.89). CONCLUSION: In our study, pharmacy supply was not associated with self-reported adherence. Most importantly, adherence and age were significant predictors of reaching undetectable viral loads.

Preliminary evidence for a human prolactin releasing factor.

Plasma from normal human subjects was subjected to a 3-phase methanol extraction. An increase in prolactin release from rat anterior pituitary tissue in vitro obtained with increasing doses of the plasma extract, while no increase in TSH release occurred. Control studies with an extract of simulated plasma using incubation medium with human serum albumin resulted in less release than a comparable plasma extract dose. The plasma extract did not lose activity with heating to 100 degrees C for 10 min, and incubation of normal plasma at 37 degrees C for 4 h prior to methanol extraction resulted in no loss of prolactin releasing activity. The plasma extract was subjected to cascade ultra-filtration through UM-10, UM-2, and UM-0.5 filters with no significant loss of prolactin releasing activity. Finally, the extract contained no TRH immunoreactivity. These results support the existence of a heat stable, ultra-filterable circulating PRF in human plasma. The origin of this material is unknown.

A survey of recovering Maryland physicians.

In a survey of physicians who had participated in the Maryland Physician Rehabilitation Program, 75 percent of the respondents reported that they were in recovery, with a mean recovery duration of eighty-eight months.

Spontaneous and antigen-induced production of HIV-inhibitory beta-chemokines are associated with AIDS-free status.

The beta-chemokines RANTES, macrophage inflammatory protein (MIP)-1alpha, and MIP-1beta suppress infection by macrophage-tropic strains of HIV and simian immunodeficiency virus (SIV) by binding and down-regulating the viral coreceptor, CCR5. Accordingly, we have examined whether higher levels of CCR5 ligands are associated with a more favorable clinical status in AIDS. A cross-sectional study of 100 subjects enrolled in the Multicenter AIDS Cohort Study at the Baltimore site was conducted to measure chemokine production and lymphocyte proliferation by peripheral blood mononuclear cells (PBMC). Statistical analyses of the data revealed that the production of HIV-suppressive beta-chemokines by HIV antigen-stimulated PBMC was significantly higher in HIV-positive subjects without AIDS compared with subjects with clinical AIDS. Increased chemokine production was also correlated with higher proliferative responses to HIV antigens. Both parameters were significantly lower in the AIDS versus non-AIDS group. Notably, significantly higher levels of MIP-1alpha were also observed with unstimulated PBMC from seronegative subjects at risk for HIV infection released as compared with seropositive and non-Multicenter AIDS Cohort Study seronegative subjects. The association of chemokine production with antigen-induced proliferative responses, more favorable clinical status in HIV infection, as well as with an uninfected status in subjects at risk for infection suggests a positive role for these molecules in controlling the natural course of HIV infection.

Geographic diversity of adult t-cell leukemia/lymphoma in Brazil. The Brazilian ATLL Study Group.

We describe 195 cases of adult T-cell leukemia/lymphoma (ATLL) reported to the national registry of T-cell malignancies in Brazil between 1994 and 1998. We compared the effect of demographic differences and clinical features of 150 consecutive ATLL cases in different regions of this diverse country. At diagnosis, the predominant clinical sub-type was the acute type (60%), followed by lymphoma (22%), chronic (10%) and smoldering (8%) types. Although we expected that different sub-types would be present in different regions, on the basis of immunogenetic factors determined by ethnicity, we did not demonstrate these differences. There were no significant differences among ATLL subtypes by age or gender. No ethnic group predominated in the total population of patients, but significant differences were noted when examining ethnic distribution by region. Reflecting the general population distribution, white patients were seen more often in São Paulo and black patients in Bahia, than in other regions. In most regions, cases were equally distributed between blacks and mulattos, except in Pernambuco, where blacks were less frequent. The main clinical features were lymphadenopathy, skin lesions, hypercalcemia and hepatomegaly. Fourteen patients (9%) suffered from HTLV-I-associated myelopathy (HAM/TSP), either at diagnosis or during follow-up of ATLL. All cases but one had antibodies to HTLV-I, with concordant results with ELISA, WB and PCR analyses. For the antibody-negative case, pol and tax gene sequences were present in tumor cells when subjected to PCR analyses. The prognosis was generally poor, suggesting that the disease in Brazil behaves in similar fashion regardless of ethnic or geographical differences.

Antibodies to the HIV-1 Tat protein correlated with nonprogression to AIDS: a rationale for the use of Tat toxoid as an HIV-1 vaccine.

OBJECTIVES: To investigate which immune parameters, such as antibodies against HIV-1 specificities, or viral parameters, such as p24 antigenemia, are predictive of disease progression. STUDY DESIGN: We performed studies on serum collected from individuals exhibiting two extremes of disease evolution--67 fast progressors (FP) and 182 nonprogressors (NP)--at their enrollment. After a 1- to 2-year clinical follow-up of 104 nonprogressors after their enrollment, we could determine the best serologic predictors for disease progression. METHODS: We investigated levels of antibodies to tetanus toxoid and to HIV antigens including Env, Gag, Nef, and Tat proteins, as well as p24 antigenemia, viremia, CD4 cell count, and interferon-alpha (IFN-alpha) titers in FPs and NPs, and we correlated these data with clinical and biologic signs of progression. RESULTS: p24 Antigenemia, a marker of viral replication, and anti-Tat antibodies were highly and inversely correlated in both groups (P < .001). Furthermore, anti-p24 antibodies and low serum IFN-alpha levels were correlated to the NP versus the FP cohort. Finally, among NPs, only antibodies to Tat and not to the other HIV specificities (Env, Nef, Gag) were significantly predictive of clinical stability during their follow-up. CONCLUSION: Antibodies toward HIV-1 Tat, which are inversely correlated to p24 antigenemia, appear as a critical marker for a lack of disease progression. This study strongly suggests that rising anti-Tat antibodies through active immunization may be beneficial in AIDS vaccine development to control viral replication.

Synchronized cyclic capillary electrophoresis using channels arranged in a triangle and low voltages.

Synchronized cyclic capillary electrophoresis (SCCE) makes use of a closed loop separation channel by which the same sample can be separated during many cycles. This enables the repeated use of the same voltage for separations such that a high total voltage, and thus high efficiency, is obtained for the synchronized components. This can be accomplished by using any type of polygon geometry for the separation channel; and calculations of the available field and number of connections needed for polygons from 3 to 5 sides are presented. Triangular designs have the advantage of using the lowest number of wells. Such designs are described, with two additional features compared to that of earlier work: 1. voltage connections that are much shallower than the separation channel, to reduce losses and dispersion at the intersections; and 2. corners that are narrower than the separation channels to reduce dispersion in the turns. Experimental data is presented for the separation of a mixture of amino acids, and for a DNA separation in a polymeric sieving matrix. The DNA separation is most sensitive to the corner dispersion problem, which reduces the observed efficiency for that separation.

Centrifugation and capillarity integrated into a multiple analyte whole blood analyser.

A unique clinical chemistry analyser is described which processes 90 mul of whole blood (fingerstick or venous) into multiple aliquots of diluted plasma and reports the results of 12 tests in 14 min. To perform a panel of tests, the operator applies the unmetered sample directly into a single use, 8 cm diameter plastic rotor which contains the required liquid diluent and dry reagents. Using centrifugal and capillary forces, the rotor meters the required amount of blood, separates the red cells, meters the plasma, meters the diluent, mixes the fluids, distributes the fluid to the reaction cuvettes and mixes the reagents and the diluted plasma in the cuvettes. The instrument monitors the reagent reactions simultaneously using nine wavelengths, calculates the results from the absorbance data, and reports the results.