Clinical outcomes and safety with trabectedin therapy in patients with advanced soft tissue sarcomas following failure of prior chemotherapy: results of a worldwide expanded access program study.

BACKGROUND: This expanded access program (EAP) was designed to provide trabectedin access for patients with incurable soft tissue sarcoma (STS) following progression of disease with standard therapy. The outcomes of trial participants accrued over approximately 5 years are reported. PATIENTS AND METHODS: Adult patients with advanced STS of multiple histologies, including leiomyosarcoma and liposarcoma (L-sarcomas), following relapse or disease progression following standard-of-care chemotherapy, were enrolled. Trabectedin treatment cycles (1.5 mg/m(2), intravenously over 24 h) were repeated q21 days. Objective response, overall survival (OS), and safety were evaluated. RESULTS: Of 1895 patients enrolled, 807 (43%) had evaluable objective response data, with stable disease reported in 343 (43%) as best response. L-sarcoma patients exhibited longer, OS compared with other histologies [16.2 months (95% confidence interval (CI) 14.1-19.5) versus 8.4 months (95% CI 7.1-10.7)], and a slightly higher objective response rate [6.9% (95% CI 4.8-9.6) versus 4.0% (95% CI 2.1-6.8)]. The median treatment duration was 70 days representing a median of three treatment cycles; 30% of patients received ≥ 6 cycles. Safety and tolerability in this EAP were consistent with prior clinical trial data. CONCLUSION: Results of this EAP are consistent with previous reports of trabectedin, demonstrating disease control despite a low incidence of objective responses in advanced STS patients after failure of standard chemotherapy. CLINICALTRIALS.GOV: NCT00210665.

Sibutramine: a new weight loss agent without evidence of the abuse potential associated with amphetamines.

Sibutramine is a serotonin and norepinephrine reuptake inhibitor that has shown efficacy as a weight loss and weight maintenance agent. Because of the abuse liability and physical dependence potential of amphetamines and related antiobesity agents, this study evaluated the abuse potential of sibutramine and compared it with that of dextroamphetamine and placebo in recreational stimulant users. Thirty-one male recreational stimulant users participated in this single-site, Latin square crossover study that compared the effects of two doses of sibutramine (20 mg and 30 mg) to dextroamphetamine (20 mg and 30 mg) and placebo, using a series of validated subjective scales or questionnaires. For scales measuring stimulation and euphoria, there was a greater mean response for dextroamphetamine 30 mg versus 20 mg, with both doses having a significantly greater stimulant and euphoric effect than placebo at the majority of time points (p < 0.05); responses for both doses of sibutramine were statistically indistinguishable from placebo at all time points. Responses to "street value" and "most enjoyed study session" questions confirmed that sibutramine lacks abuse potential; mean cash value estimates of street value were significantly greater for both dextroamphetamine doses than for placebo or either sibutramine doses (p < 0.05), and the rank order of session enjoyment placed both doses of sibutramine last. Together with the relatively late Tmax of the active metabolites (3-4 hours), this short-term, single-dose study provides strong evidence that sibutramine does not have the potential for abuse that is characteristic of amphetamines and that it is indistinguishable from placebo in abuse potential.

Analysis of the Anopheles (Anopheles) quadrimaculatus complex of sibling species (Diptera: Culicidae) using morphological, cytological, molecular, genetic, biochemical, and ecological techniques in an integrated approach.

The Anopheles quadrimaculatus complex of 5 cryptic species (i.e., An. diluvialis Reinert, new species; An. inundatus Reinert, new species; An. maverlius Reinert, new species; An. quadrimaculatus Say; An. smaragdinus Reinert, new species) is analyzed using multiple techniques, including morphological, cytological, molecular, genetic, biochemical, and ecological procedures. All life stages (egg, 4th-instar larva, pupa, and female and male adults) are described using morphological features, and pertinent stages or structures are illustrated. A neotype for An. quadrimaculatus is designated, and the synonymy of An. annulimanus Van der Wulp is confirmed. Several new morphological features are described. New and summarized data from published literature on hybridization, cytological, electrophoretic, molecular, and cuticular hydrocarbon studies are included. Immature and adult bionomics are given. The geographic distribution for each species is listed and shown on maps. Procedures for collecting, processing, and rearing specimens are described. Keys using morphological characters are included for the eggs, 4th-instar larvae, pupae, adult females, and male genitalia. Also, a biochemical key for the 5 species is included. Color and pattern variations of larvae and pupae are discussed.

Human blood-feeding rates among sympatric sibling species of Anopheles quadrimaculatus mosquitoes in northern Florida.

We compared rates of feeding on human hosts for blood-engorged female Anopheles quadrimaculatus species A, B and C1 collected from daytime resting sites in Manatee Springs State Park, Levy County, Florida during 1992-1993. Quick-blot DNA probes were used to identify mosquito taxa and also the presence of human blood in the mosquito gut. In collections from a campground area, human blood-feeding rates differed significantly among mosquito species (10.7% [19 of 177], 0%, [0 of 62], and 1.2%, [4 of 327]), respectively for species A, B and C1). In collections from a woodland site (1 km from the campground), 1.5% (2 of 129) of the species B females had fed on humans, whereas none of 19 species A or 159 species C1 females had done so. Of the three species in this study area, species A appears the most likely to be a biting pest of humans and a vector of human malaria.

Anopheles perplexens from artificial containers and intermittently flooded swamps in northern Florida.

Anopheles perplexens was collected from habitats previously unreported for this species in northern Florida. These habitats included intermittently flooded swamps, water-filled tires, and plastic oviposition cups. First-instar An. perplexens larvae were recovered from soil samples collected in an intermittently flooded swamp that were flooded in the laboratory, suggesting that An. perplexens eggs may survive in the soil during dry periods. Anopheles perplexens larvae were collected from water-filled tires and plastic oviposition cups at sites near Gainesville, FL. Of 30 dissected An. perplexens females collected in updraft CDC traps, all had ovarioles in Christophers's stage II and blood was absent in the midgut. The physiological state of these females indicates that updraft CDC traps collect predominately host-seeking females and that females take one blood meal per gonotrophic cycle.

Anopheles perplexens from artificial containers and intermittently flooded swamps in northern Florida

Anopheles perplexens was collected from habitats previously unreported for this species in northern Florida. These habitats included intermittently flooded swamps, water - filled tires, and plastic oviposition cups. First - instar An. perplexens larvae were recovered from soil samples collected in an intermittently flooded swamp that were flooded in the laboratory, suggesting that An. perplexens eggs may survive in the soil during dry periods. Anopheles perplexens larvae were collected from water-filled tires and plastic oviposition cups at sites near Gainesville (AU)

Multiple-technique identification of sibling species of the Anopheles quadrimaculatus complex.

In the past, most researchers used a single technique for identification of cryptic taxa, population structures, biosystematics, and phylogenetic studies. Our experience with the Anopheles quadrimaculatus complex shows the importance of using several methods on individual mosquitoes. This approach consists of analysis of the polytene chromosomes in ovarian nurse cells, gas chromatographic profiles of cuticular hydrocarbons, isozyme electrophoresis, and restriction site analysis of mitochondrial or genomic DNA. We recommend use of this multiple-technique approach when analyzing feral populations for the first time, or for correlating information obtained by investigators using different techniques.

Short-term changes in the abundance and parity rate of Anopheles quadrimaculatus species C (Diptera: Culicidae) in a central Florida swamp.

Updraft CDC traps baited with dry ice were used to monitor changes in the abundance and parity rate of Anopheles quadrimaculatus Say complex mosquitoes in an intermittently flooded swamp in Central Florida during an 18-d period. Mosquitoes collected each day were identified to species using DNA hybridization and isozyme electrophoretic techniques and were dissected to determine follicular maturation and parity. Of 1,178 An. quadrimaculatus mosquitoes identified to species, 4% were species A and 96% were species C. Dissections of females of both species indicated that 98% were nonblood fed and nongravid with ovariole development at Christophers' stage II. Overall parity rates were 0.19 and 0.51 for populations of species A and species C, respectively. The duration of the gonotrophic cycle for species C females was estimated to be 5 d based on the interval between peaks in the number of nulliparous females collected on days 1 and 6 and peaks in the number of parous females collected 5 d later. Significant trends were observed in the number of parous species C females collected each day, indicating that the parity rate could not be used as an unbiased estimator of survivorship.

Comparative susceptibility of species A, B and C of Anopheles quadrimaculatus complex to infection with subperiodic Brugia malayi and Brugia pahangi (Nematoda: Filarioidea).

Susceptibilities of natural populations of sibling species A, B and C of the Anopheles quadrimaculatus complex and the colonized strain A to subperiodic Brugia malayi and Brugia pahangi were compared. All 3 sibling species showed varying degrees of susceptibility to both B. pahangi and B. malayi, and they were considerably more susceptible to B. pahangi than to B. malayi. The rate and intensity of infection to B. pahangi were highest for species A (66.2% and 7.4 L3/female, respectively) and lowest for species B (21.3% and 1.7 L3/female). For B. malayi these values were higher for species A (29.7% and 1.84 L3/female) than for species B (13.3% and 0.86 L3/female) and C (12.6% and 0.75 L3/female). The colonized strain A of An. quadrimaculatus was significantly more susceptible to both Brugia species than the natural populations of sibling species A, B and C.

Evidence for microgeographic genetic subdivision of Anopheles quadrimaculatus species C.

Species C of the Anopheles quadrimaculatus complex has a patchy distribution and has been found principally near the coast in river and springs systems of northwest Florida (GULF, SR, SFR--designated as C1, and CHOC--designated as C2) and the Ogeechee River (OGE--C2) near Savannah, Georgia. We have documented significant differences in allelic frequencies at 5 loci (malic enzyme, aconitase, aldehyde oxidase, esterase-5 and esterase-6), and genetic substructuring (Fst) between C1 and C2. The GULF and CHOC populations occurred in apparently similar environments, but were separated by about 160 km. Both these populations are separated from OGE by over 160 km (with low to undetectable distribution of species C in the transect regions). The higher genetic similarity of the CHOC with OGE populations indicated either selective pressures or genetic drift or both as being responsible for the differences between C1 and C2 populations.

Identification of species D, a new member of the Anopheles quadrimaculatus species complex: a biochemical key.

Sibling species D, a new member of the Anopheles quadrimaculatus species complex was identified in collections from Pickwick Lake, Tishomingo County, Mississippi and Choctawhatchee, Bay County, in West Florida. This species occurred sympatrically with the previously described species, A, B and C. Evidence for identification of species D includes diagnostic allozymes, a lack of polytene chromosomes in the ovarian nurse cells, and inviability of F1 progeny and lack of sperm transfer in hybridization crosses. An electrophoretic taxonomic key for distinguishing species D from A, B and C is presented.

Dichotomous electrophoretic taxonomic key for identification of sibling species A, B, and C of the Anopheles quadrimaculatus complex (Diptera: Culicidae).

Samples of 17 populations of Anopheles quadrimaculatus Say from Florida, Alabama, Arkansas, Louisiana, Mississippi, Tennessee, New York, and New Jersey were analyzed for genetic variability at 33 enzyme loci. Statistical analysis of electromorph frequency distributions indicated that sympatric sibling (morphologically indistinguishable) species occurred in about 59% of the populations tested. The association of polytene chromosome and electrophoretic patterns of individual field-collected females confirmed species-specific diagnostic allozymes, which were useful in identifying sibling species A, B, and C and in estimating the proportions of each species at the 17 collection sites. A dichotomous electrophoretic key is presented for the identification of sibling species of the An. quadrimaculatus complex. The electrophoretic method is better than the ovarian polytene chromosome method, because mosquitoes of both sexes and females irrespective of their gonotrophic condition can be identified.

A new member of the Anopheles quadrimaculatus complex, species C.

A new member, species C, of the Anopheles quadrimaculatus complex was recently found in collections from the northwest coast of Florida. This new species cannot be differentiated from the other 2 species with available taxonomic keys. Evidence for this taxon as a sibling species includes data on hybrid sterility and distorted sex ratios in the progeny of crosses to species A and B, chromosomal differences, and diagnostic allozymes.

Hybridization of laboratory strains of sibling species A and B of Anopheles quadrimaculatus.

Adult mosquitoes of the Anopheles quadrimaculatus complex were collected from Montgomery County, AL (MON) and Alachua County, FL, (KBG) and laboratory stocks of species A and B were established through a selection procedure employing isofemale lines. Progeny from a cross of species B females to ORL males were usually semisterile females and sterile males. Progeny of the reciprocal cross were also semisterile females and sterile males, but the sex ratio was variable and ranged from normal to no males because of male mortality during the pupal stage. Conspecific crosses between strains from the two locations resulted in fertile offspring. Crosses between the sibling species from the two locations invariably gave semisterile females and sterile males (or lethal effects). This evidence confirmed previous data from hybridization and electrophoretic analyses of field populations indicating that Anopheles quadrimaculatus is a species complex.

The ovarian nurse cell polytene chromosomes of Anopheles quadrimaculatus, species A.

Nurse cells in the ovaries of adults of Anopheles quadrimaculatus, Species A, were used to prepare a polytene chromosome map. The chromosome quality is superior to that of salivary glands, and it is easier to use adults rather than larvae for cytological analysis of field populations. The most reliable homologies between the salivary and ovarian maps are located in the distal ends of the respective arms, and one homologous region is a prominent landmark in all of the members of the nearctic Maculipennis complex and related species. The left arm of chromosome 3 is uniquely dimorphic. The homokaryotype for 3L1 is synonymous with 3L of the published map of salivary gland polytenes. The 3L heterokaryotype is mostly asynaptic, except for two small homologous, synaptic areas, one of which is inverted. Each homokaryotype contains a unique, diffuse puff that is adjacent to the centromere.

A genetic sexing strain of Anopheles quadrimaculatus, species A.

A genetic sexing strain of a mosquito, Anopheles quadrimaculatus, Species A, was synthesized for the preferential elimination of females during the egg stage. Malathion susceptibility was used as a conditional lethal, and the dominant malathion-resistance allele was linked to the Y chromosome via a radiation-induced reciprocal translocation involving the terminal end of the right arm of chromosome 3 and the Y chromosome. Examination of mitotic chromosomes and salivary polytene chromosomes revealed the precise nature of the translocation. Genetic leakage, through recombination, in the strain was very low (0.02%).