Antiprotozoal activities of some constituents of Markhamia tomentosa (Bignoniaceae).

Phytochemical investigation of an ethyl-acetate extract of the stem bark of Markhamia tomentosa (Bignoniaceae), which had good antimalarial activity in vitro, resulted in the isolation of eight known compounds: 2-acetylnaphtho[2,3-b]furan-4,9-dione (1), 2-acetyl-6-methoxynaphtho[2,3-b]furan-4,9-dione (2), oleanolic acid (3), pomolic acid (4), 3-acetylpomolic acid (5), tormentic acid (6), beta-sitosterol (7) and beta-sitosterol-3-O-beta-D-glucopyranoside (8). The structures of these compounds were established by spectroscopic methods. Each of compounds 1, 2, 4 and 5 was evaluated in vitro for its antiprotozoal activities against the ring stages of two chloroquine-resistant strains of Plasmodium falciparum (K1 and W2), the amastigotes of Leishmania donovani, and the bloodstream trypomastigotes of Trypanosoma brucei rhodesiense (the species responsible for human malaria, visceral leishmaniasis and African trypanosomiasis, respectively). Although compounds 1 and 2 exhibited potent antiprotozoal activities, they also showed high toxicity against a mammalian (L-6) cell line.

In vitro antiprotozoal activities and cytotoxicity of some selected Cameroonian medicinal plants.

Eight extracts from seven selected Cameroonian medicinal plants, traditionally used to treat malaria and other protozoal diseases, were tested in vitro for their antiprotozoal activities against Plasmodium falciparum K1 chloroquine-resistant strain, Leishmania donovani, Trypanosoma cruzi and Trypanosoma brucei rhodesiense, protozoa responsible for malaria, visceral leishmaniasis, Chagas disease and African trypanosomiasis, respectively. The most active extract against Plasmodium falciparum K1 strain and Trypanosoma brucei rhodesiense was the methanolic extract of Albizia zygia (Fabaceae) stem bark with IC(50) values of 1.0 microg/ml and 0.2 microg/ml, respectively. Five extracts showed IC(50) values below 5mug/ml against Leishmania donovani, with the methanolic seed extract of Harungana madagascarensis showing the highest activity, but only the methanolic extract of Albizia zygia showed activity against Trypanosoma cruzi. Cytotoxicity and selectivity indexes were estimated for the most active extracts. The best ratio of cytotoxicity to antiplasmodial activity (SI(a)=14) was established for the methanolic leaf extract of Symphonia globulifera (Clusiaceae), while the methanolic stem bark extract of Albizia zygia showed the best ratio of cytotoxicity to antitrypanosomal activity (SI(b)=22.5).

Comparative antiplasmodial, leishmanicidal and antitrypanosomal activities of several biflavonoids.

The antiplasmodial, leishmanicidal and antitrypanosomal activities of eight natural biflavonoids were estimated in vitro on a chloroquine-resistant strain of Plasmodium falciparum, axenically grown Leishmania donovani amastigotes and Trypanosoma cruzi trypomastigotes and Trypanosoma brucei rhodesiense bloodstream forms. Lanaroflavone showed the highest antiplasmodial activity (IC(50) = 0.48 microM), isoginkgetin was the most active leishmanicidal compound (IC(50) = 1.9 microM), whereas ginkgetin (IC(50) = 11 microM) and isoginkgetin (IC(50) = 13 microM) showed the best antitrypanosomal activity in our assays. The cytotoxicity and the selectivity indices for the most active compounds were also estimated. Lanaroflavone exhibited a high selectivity index value (SI = 159), indicating selective antiplasmodial activity.

A bioactive biflavonoid from Campnosperma panamense.

Lanaroflavone (1), a biflavonoid isolated from the methanol extract of the aerial part of Campnosperma panamense by bioguided fractionation, has been assessed for in vitro antiprotozoal activity. Lanaroflavone showed both antimalarial and leishmanicidal activities, but was inactive against Chagas disease vector, Trypanosoma cruzi.

Combination vaccines: postlicensure safety evaluation.

The success of immunizations in nearly eliminating many vaccine-preventable diseases has resulted in an increase in the need to study risks from vaccines, combination or otherwise. The well-known limitations associated with prelicensure trials have led many to hope that postlicensure studies can address safety issues. This article reviews measures that have been or should be taken to meet this expectation: establishment of clinical immunization safety assessment centers, standardization of case definitions for vaccine adverse events, use of the Vaccine Identification Standards Initiative to improve the accuracy and efficiency with which vaccination records are transferred, integration of vaccine safety monitoring into immunization registries, establishment (and enlargement) of the Vaccine Safety Datalink project, use of innovative analytic tools for better signal detection, and implementation of various methods to overcome confounding by contraindication. Only by investing in vaccine safety infrastructure at a level commensurate with investments in vaccine development can we hope to retain the public's confidence in immunization.

"Reverse engineering" a formulary selection algorithm to determine the economic value of pentavalent and hexavalent combination vaccines.

INTRODUCTION: Combination vaccines with overlapping, noncomplementary components are being introduced to reduce the number of separate injections required to immunize children. A vaccine selection algorithm using operations research techniques was developed as a tool for vaccine purchasers to assemble formularies of monovalent and combination vaccines that would satisfy the recommended immunization schedule. The algorithm weighs distinguishing features of economic consequence among competing vaccines to achieve the lowest overall cost to payers and/or to society for immunization. This method was adapted here to solve for the purchase price of several hypothetical future pentavalent and hexavalent combination vaccines that would permit each to "win" a place in such a lowest cost formulary. METHODS: Integer programming and an iterative bisection search method determined the maximum "inclusion price" of 4 vaccines not licensed in the United States as of September, 2001 [diphtheria-tetanus-acellular pertussis (DTPa)-Haemophilus influenzae type b (HIB)-hepatitis B (HBV), DTPa-HIB-inactivated polio vaccine (IPV), DTPa-HBV-IPV and DTPa-HIB-HBV-IPV], in competition with 15 existing formulations of licensed vaccines for these diseases at their March, 2000, federal contract discount prices. Both 5-visit and 6-visit scenarios were studied. Different preparation costs were assigned to lyophilized powder ($1.50), liquid ($0.75) and prefilled-syringe ($0.25) formulations/packaging. Injection costs were varied stepwise from $5 through $45 for each dose administered, shifting from a payer's to a societal perspective. RESULTS: Overall inclusion prices (maximum price for each candidate vaccine to be included in a lowest cost formulary) ranged from $9 to $129 per dose depending on cost assumptions and usage frequency (values would be higher if competing against private-sector vaccine prices). The range was $27 to $68 per dose for DTPa-HIB-HBV, at optimal utilization to avoid extra vaccination. Similarly, as injection costs varied from $5 to $45, DTPa-HIB-IPV ranged from $28 to $75. With the same assumptions, DTPa-HBV-IPV would earn a place in a best value formulary at prices from $35 to $76. As expected the inclusion prices for hexavalent DTPa-HIB-HBV-IPV, $40 to $123, were higher (reflecting more economic value) than for pentavalents. When the assumed injection costs rose to > or = $8, the more expensive HIB-HBV and DTPa-HIB tended to appear in lowest cost formularies, because their cost premium over separate monovalent and trivalent products was outweighed by the savings from one fewer injection. CONCLUSION: Reverse engineering the vaccine selection algorithm provides a tool to demonstrate the economic value of new combination vaccines and to make pricing decisions.

Economic value to parents of reducing the pain and emotional distress of childhood vaccine injections.

BACKGROUND: One reason that recommended childhood immunizations due at child health visits are deferred is to avoid the pain and emotional distress associated with the increasing number of injections required. This deferral leads to additional visits and costs and reduced immunoprotection against vaccine-preventable illnesses. To assess the economic value of combination vaccines that address this problem, we surveyed parents to determine the amount they would be willing to pay to avoid the pain and emotional distress experienced by their infants from injections. METHODS: A self-administered questionnaire was completed within 24 h of the vaccinations by 294 parents of children ages 11/2 to 7 months receiving vaccine injections at 26 outpatient child health centers. The willingness-to-pay (WTP) method was used to estimate the intangible cost of the pain and emotional distress of the 1 to 4 injections their child had received. Parents were asked how much of their own money they would have paid to avoid these injections, without any compromise in the safety and efficacy of the vaccinations. RESULTS: Wide variations in WTP amounts were observed, ranging from median values of $10 to $25 and average values of $57.06 to $79.28 to avoid the pain and emotional distress associated with eliminating all injections at visits in which one to four injections were administered. Parents placed greater value on reductions that avoided all injections than on reductions that avoided only some injections. Overall the median cost per injection avoided was $8.14, and the mean was $30.28. CONCLUSIONS: Parents have strong preferences for limiting vaccine injections. The economic cost of the pain and distress associated with such injections, reflected in the amounts they report they would be willing to pay to avoid them, represents a substantial component of the cost of disease control through immunization.

Quantitative determination of naphthoquinones of Impatiens species.

A convenient and reliable reversed phase-HPLC method has been developed and validated for the quantitative determination of naphthoquinones present in the aerial parts of Impatiens glandulifera (Balsaminaceae) during two growing seasons (May to August 1998 and June to October 1999). Maximal content (0.8-1.1%) of 2-hydroxy-1,4-naphthoquinone and its 2-methylated derivative was observed in flowers collected between July and August. The procedure was applied to compare the pigment content in three other species of Impatiens but showed 2.5-37 times lower levels than those found in I. glandulifera.

Antiprotozoal activities of Colombian plants.

In our search for therapeutical alternatives for antiprotozoal chemotherapy, we collected a selection of 44 plants from western Colombia upon ethnopharmacological and chemotaxonomic considerations. Polar and apolar extracts of these species were examined for antimalarial activity using in vitro tests with two clones of Plasmodium falciparum. Leishmanicidal and trypanocidal activity were determined in vitro using promastigote and amastigote forms of several strains of Leishmania sp. and epimastigotes of Trypanosoma cruzi. Among the selected plants, the 15 following species showed good or very good antiprotozoal activity in vitro: Aspidosperma megalocarpon, Campnosperma panamense, Conobea scoparioides, Guarea polymera, Guarea guidonia, Guatteria amplifolia, Huberodendron patinoi, Hygrophila guianensis, Jacaranda caucana, Marila laxiflora, Otoba novogranatensis, Otoba parviflora, Protium amplium, Swinglea glutinosa and Tabernaemontana obliqua. Cytotoxicity was assessed in U-937 cells and the ratio of cytotoxicity to antiprotozoal activity was determined for the active extracts. Ten extracts from eight species showed selectivity indexes > or = 10. Among the extracts that showed leishmanicidal activity, the methylene chloride extract of leaves from C. scoparioides showed a selectivity index in the same range that the one of the Glucantime control. Several of the active leishmanicidal plants are traditionally used against leishmaniasis by the population of the concerned area.

Safety and immunogenicity of varying dosages of trivalent inactivated influenza vaccine administered by needle-free jet injectors.

To evaluate the perceived pain, other adverse events, and immunogenicity of influenza virus vaccine administered by needle-free jet injector (JI) compared with that of vaccine administered by needle and syringe (N&S), we randomly assigned 304 healthy young adults to receive one of three dosages (0.5, 0.3, or 0.2 ml) of the 1998-1999 season vaccine administered by either of two JI devices or by N&S. In multivariate analysis, female gender and JI administration were associated with higher levels of pain reported at the time of vaccination as well as with the occurrence of local injection site reactions following vaccination. Immune response did not vary significantly by dosage but administration by one JI device was associated with higher post-vaccination H1N1 antibody titers.

Bioactive acridone alkaloids from Swinglea glutinosa.

A new prenylated acridone alkaloid, 1,3,5-trihydroxy-2,8-bis(3-methylbut-2-enyl)-10-methyl-9-acridone (1), was isolated from the stembark of Swinglea glutinosa, along with three known acridone alkaloids, 5-hydroxynoracronycine (2), 1,3,5-trihydroxy-4-methoxy-2-(3-methylbut-2-enyl)-10-methyl-9-acridone (3), and 1,3,5-trihydroxy-4-methoxy-10-methylacridone (4). The isolated alkaloids were assessed in vitro against chloroquine-sensitive and -resistant Plasmodium falciparum strains and for cytotoxicity using HeLa cells.

Saponins from Strasburgeria robusta.

The isolation of three saponins, 24-hydroxytormentic acid ester glucoside (1), niga-ichigoside F1 (2) and niga-ichigoside F2 (3), from the stem bark of Strasburgeria robusta, an endemic plant from New Caledonia, is reported.

Improved procedure for the quality control of Hypericum perforatum L.

A new, fast and reliable procedure for the quantification of the major compounds of Hypericum perforatum L. has been developed. Four naphthodianthrones (protopseudohypericin, pseudohypericin, protohypericin, hypericin) and two phloroglucinols (hyperforin, adhyperforin) were assayed by HPLC using a short (17 min) linear gradient, with hypericin and hyperforin as external standards. Extraction of dried plant material with methanol in the dark at room temperature for 2 h led to a complete recovery of phloroglucinols but only a partial recovery of the naphthodianthrone derivatives. Treatment of plant material with water:ethanol (40:60, v/v) in a water bath shaker at 80 degrees C led to the total extraction of hypericins, but a 10% loss of total hyperforins was also observed. The two extraction methods, applied successively to the same sample, allowed the complete extraction of all compounds of interest. A 5 min exposure of the crude extract of H. perforatum to sunlight (1 E/m2) induced a 96% loss of hyperforins, whereas the dry plant material lost only 20% of hyperforins after 2 h exposure to sunlight (24 E/m2).

Triterpenoid saponins from Isertia pittieri.

Two new 27-nor-triterpenoid saponins, pyrocincholic acid 3 beta-O-beta-D-quinovopyranosyl-28-[beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl] ester (1) and pyrocincholic acid 3 beta-O-beta-D-quinovopyranosyl(1-->6)-beta-D-glucopyranosyl-28-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl] ester (2) were isolated from the stem bark of Isertia pittieri, together with two known bidesmosidic quinovic acid glycosides. The structures of 1 and 2 were determined on the basis of spectroscopic studies.

Actividad antipacudica y potenciación de lacloroquina con alcaloides aporfinicos in vitro / In vitro antimalarial activity and chloroquine potentiating action of aporphine alkaloids

Pregunta de investigación: ¿Los alcaloides aporfínicos tiene una actividad intrinseca antipalúdica, pueden potencializar la acitivad de la cloroquina y revertir la resitencia de Plasmodium falciparum?. OBjetivos: Determinar la actividad antipaludica intrinseca de 12 alcaloides aporfínicos, Determinar el nivel de su efecto, establecer si tiene efecto y de maduracion o ponteciación, sinergismo aditivo o de antagonismo con la cloroquina, Determinar qué alcaloide pueden provocar reversión de la resistencia a la cloroquina, Estudiar un posible mecanismo de acción. Lugar:IINSAD, IBBA. Métodos: Cultivo de estadios eritrocitarios de Plasmodium falciparum, evaluación de la actividad antipalúdica in vitro de los alcaloides aporfinicos usando varios métodos. Resutlados: Los alcaloides evaluados son menos acativos que la cloroquia sola, todos los alcaloides mostraron un claro efecto acumulativo, especialmente Glaucina fumarato, Los alcaloides asimilobina, isoboldina HCI, coridina y actinodafnia conbinados con lacloroquina mostraron un efecto antagónico hacia esta droga, losalcaloides nuciferina HCI, pachyconfina, cassiticina, presentan un efecto de potenciación a la cloroquina. Glaucina fumarato, lautotetanina e isocorytuberina mostraron un sinergismo aditivo a la cloroquina, Nuceferina HCI revierte la resisstencia de la cloroquina a 0.5ug/ml, pachyconfina a la concentraion de 2 ug/ml y cassyticina a 1 ug/ml, se evidenció que los alcaloides aorfinicos no perturban el mecanismo de transporte creado por los parásitos intraeritrocitarios. Conclusión: Los 12 alcaloides aporfinicos, no presentan una actividad mayor que la cloroquina, pero nuciferina HCI, pachyconfina y cassyticina convinados con esta 4 aminoquinolina, pueden potencializar su actividad y revertir la resitencia de cepas cloroquina-resistentes, la modalidad de acción de estos alcaloides es de tipo acumulativo y no perturban el mecanismo de transporte creado por los parásitos intraeritrocitarios.

The hidden costs of infant vaccination.

Combination vaccines to minimize injections required for infant vaccination, and new vaccines with improved safety profiles, will pose increasingly complex choices for vaccine purchasers in the future. How much of a premium to pay for such vaccines might be determined by taking into account (1) the psychological burden of multiple injections during a single clinic visit, and the costs of any additional visits to minimize these, and (2) the medical, work-loss, and incidental costs of common vaccine-associated symptoms. This cross-sectional survey included randomly-selected parents of 1-8-month-old infants who received vaccines in a Northern California health maintenance organization (HMO) in 1997. Interviewers called parents 14 days after the infant's vaccination to administer a 10-minute closed-ended interview in English or Spanish. Parents were asked about infant symptoms after vaccination, their preferences regarding multiple injections and their (theoretical) willingness to pay to reduce the number of injections their infant would receive, or to avoid the adverse symptoms experienced. Among 1769 eligible infants, interviews were completed with parents of 1657 (93%). The psychological cost of multiple injections was estimated by the willingness of parents to pay a median of $25 to reduce injections from 4 to 3, $25 from 3 to 2, and $50 from 2 to 1. Vaccine-associated symptoms caused mean costs of $42 in medical utilization and $192 in work-loss among the families who experienced those events (Ns=62 and 35, respectively). When averaged among all 1657 study infants, vaccine-associated symptoms after the index vaccination visit resulted in $2.91 in medical utilization, $4.05 in work-loss, and $0.74 in direct nonmedical costs, yielding total financial costs of $7.70. Parents of infants who had vaccine-associated symptoms said they would have paid a median of $50 to avoid these symptoms. Fever and fussiness were associated in logistic regression analysis with a two-fold increase in the odds of medical utilization, and fever with more than a three-fold increase in work loss. We conclude that multiple injections during a single clinic visit entail psychological costs. The psychological costs of vaccine-associated symptoms, as measured by willingness-to-pay methods, are higher than those resulting from multiple injections. The financial costs of medical utilization and work-loss resulting from common vaccine-associated symptoms are non-negligible and should be incorporated in economic analyses.

Safety testing of needle free, jet injection devices to detect contamination with blood and other tissue fluids.

Needle free jet injection guns have been used extensively in both veterinary and human health to deliver both vaccine and drugs, but in recent years, concerns have mounted for their potential to transmit blood borne disease agents among consecutive vaccinates. A Ped-O-Jet type jet injection device was used to deliver serial subcutaneous injections of 0.5 mL saline (as a surrogate for vaccine) into calves and pigs, with intervening ejectates collected in vials to represent what the next vaccinate would have received. An enzyme linked immunosorbant assay was developed to detect species specific albumin as a marker for blood, using calibration standards from known dilutions of bovine or porcine blood. Assay sensitivity of 20 pL/mL corresponded to the estimated minimal chimpanzee infectious dose of 10 pL for hepatitis B virus. The methodology and available results for evaluating the safety of jet injector devices are reported.

HIV-1 and other sexually transmitted infections in a cohort of female sex workers in Chiang Rai, Thailand.

OBJECTIVES: To determine demographic and behavioural factors and sexually transmitted infections (STIs) associated with prevalent HIV-1 infection among brothel based and other female sex workers (FSWs) in Chiang Rai, northern Thailand. METHODS: Data were collected from questionnaires, physical examinations, and laboratory evaluations on Thai FSWs enrolled in a prospective cohort study in Chiang Rai, Thailand, from 1991 to the end of 1994. RESULTS: HIV-1 seroprevalence was 32% among 500 women: 47% for 280 brothel workers and 13% for 220 other FSWs (p < 0.001); 96% of infections were due to HIV-1 subtype E. At enrolment, other STIs were common: chlamydia, 20%; gonorrhoea, 15%; active syphilis (serological diagnosis), 9%; genital ulcer, 12%; seroreactivity to Haemophilus ducreyi, 21%, and herpes simplex virus type 2 (HSV-2), 76%. On multiple logistic regression analysis, HIV-1 was associated with brothel work, birth in upper northern Thailand, initiation of commercial sex at < 15 years of age, syphilis, HSV-2 seropositivity, and genital ulcer. CONCLUSIONS: Young Thai FSWs working in brothels in northern Thailand in the early phase of the HIV epidemic have been at very high risk for HIV-1 infection and several other STIs. Programmes are needed to prevent girls and young women from entering the sex industry and to reduce the risk of infection with HIV-1 and other STIs.