Phytoseiidae (Acari: Mesostigmata) on plants of the family Solanaceae: results of a survey in the south of France and a review of world biodiversity.

Species of the family Phytoseiidae are predators of pest mites and small insects. Their biodiversity is not equally known according to regions and supporting plants. This paper focuses on Phytoseiidae species on plants of the family Solanaceae. The Solanaceae contain many cultivated plants, for example tomato on which leaf characteristics hinder Phytoseiidae settlement and dispersal. This study presents (i) results of surveys carried out on Solanaceae in the south of France, and (ii) Phytoseiidae biodiversity on Solanaceae worldwide. Eleven species were retrieved on 20 solanaceous plants in the south of France with four main species: Euseius gallicus, Euseius stipulatus, Phytoseiulus persimilis and Typhlodromus (Anthoseius) recki. The global analysis suggests that much more species might be found enhancing sampling efforts, whatever the biogeographic region considered. Five Phytoseiidae genera concentrate the highest number of reports and species [Amblyseius, Neoseiulus, Euseius, Phytoseius and Typhlodromus (Anthoseius)]. These genera are not evolutionarily related; adaptation on Solanaceae seems to be recent, except in the Neotropical region. The latter region represents the highest number of reports, species and Solanaceae plants sampled, probably as the centre of origin of this plant family. Occurrence probabilities in biogeographic regions and plant genera are provided as a baseline for searching for new predators adapted to Solanaceae.

Actively personalized cancer vaccines--the step into clinical application.

Cancer vaccine development enters a new phase of innovation based on the development of modern sequencing technologies and novel RNA-based synthetic drug formats which enable the analysis and therapeutic targeting of every patient's tumor genome. By applying and combining these innovations, we have brought the concept of "actively personalized cancer vaccines" to clinical testing. Synthetic RNA is used as the drug format, allowing affordable, individual "on demand" manufacturing of tumor-optimized vaccines.

Translation of genomics-guided RNA-based personalised cancer vaccines: towards the bedside.

Cancer is a disease caused by DNA mutations. Cancer therapies targeting defined functional mutations have shown clinical benefit. However, as 95% of the mutations in a tumour are unique to that single patient and only a small number of mutations are shared between patients, the addressed medical need is modest. A rapidly determined patient-specific tumour mutation pattern combined with a flexible mutation-targeting drug platform could generate a mutation-targeting individualised therapy, which would benefit each single patient. Next-generation sequencing enables the rapid identification of somatic mutations in individual tumours (the mutanome). Immunoinformatics enables predictions of mutation immunogenicity. Mutation-targeting RNA-based vaccines can be rapidly and affordably synthesised as custom GMP drug products. Integration of these cutting-edge technologies into a clinically applicable process holds the promise of a disruptive innovation benefiting cancer patients. Here, we describe our translation of the individualised RNA-based cancer vaccine concept into clinic trials.

Different feeding behaviors in a single predatory mite species. 1. Comparative life histories of three populations of Phytoseiulus longipes (Acari: Phytoseiidae) depending on prey species and plant substrate.

The spider mites Tetranychus evansi and T. urticae are key pests of tomato crops, for which no sustainable practical control strategy is available yet. A Brazilian (B) and an Argentinean (A) population of a phytoseiid predatory mite species, Phytoseiulus longipes, are able to develop and reproduce on T. evansi on tomato, whereas a Chilean (C) population is not. In order to better characterize the two distinct feeding behaviours of these three populations, life table data were assessed when the predator was offered T. evansi or T. urticae as prey on bean or tomato leaves. No effect of the prey offered nor the plant substrate was demonstrated on development durations of the three populations. However, immature mortality was low for the Argentinean and the Brazilian populations whatever the prey or plant substrate, whereas 89 % of P. longipes from Chile died before reaching adulthood when fed T. evansi on tomato. No difference in effect on female longevity was detected among the three populations. Finally, the demographic parameters of all populations were lower in presence of tomato compared to beans. Possible explanations for these results are discussed.

Different feeding behaviours in a single predatory mite species. 2. Responses of two populations of Phytoseiulus longipes (Acari: Phytoseiidae) to various prey species, prey stages and plant substrates.

Tritrophic studies involving several populations of the predatory mite Phytoseiulus longipes showed distinct life history traits depending on the prey offered and/or the plant substrate. In order to better understand the biology of this predator, the response to several combinations of prey species (Tetranychus evansi and Tetranychus urticae), prey stages (eggs and mobile stages) and plant substrates (bean and tomato leaf discs) has been assessed for two populations of P. longipes. Unlike what was found for life history traits, both populations displayed similar behaviour: they went and stayed more on tomato than on bean, they preferred T. urticae over T. evansi and mobile stages over eggs. Combining the previous life history data with the present results suggests that P. longipes may display host-plant mediated specialization on a prey species, T. evansi. Possible underlying mechanisms are discussed, as well as directions for future work.

Obtaining molecular data for all life stages of Typhlodromus (Typhlodromus) exhilaratus (Mesostigmata: Phytoseiidae): consequences for species identification.

Several species of the family Phytoseiidae are known to control mite pests in many crops worldwide. However, biological control success greatly depends on the accurate identification of these predatory mites. Species diagnostics is essentially based on the morphological characters of females. Thus, when only immature stages and/or males are collected, their identification is poorly supported. Molecular tools could be of great help to overcome these difficulties, as molecular sequences are assumed to be identical for the life stage considered. However, one of the essential points is to extract a sufficient DNA amount from a single specimen of immature stages (eggs, protonymphs, deutonymphs) and males (less than 300 µm in length) to amplify and sequence DNA. The markers used were two mitochondrial DNA fragments (12S rRNA and Cytb mtDNA) and the species studied were Typhlodromus (Typhlodromus) exhilaratus and T. (T.) phialatus, two cryptic species, reported to control mite pests in crops of southern Europe and commonly found on the same plants. Despite a low quantity of DNA extracted, particularly for the egg, larva and protonymph stages, DNA was amplified and sequences were obtained from all the life stages considered with the two mtDNA fragments. Sequences from all the developmental stages of T. (T.) exhilaratus were identical and well differentiated from those of its sister-species. However, contaminations were observed especially for eggs and DNA amplified with the Cytb mt marker. Utility of the present results are discussed and protocol improvements are proposed.

Selective uptake of naked vaccine RNA by dendritic cells is driven by macropinocytosis and abrogated upon DC maturation.

Even though it is known for more than one decade that antigen-encoding RNA can deliver antigenic information to induce antigen-specific immunity against cancer, the nature and mechanism of RNA uptake have remained enigmatic. In this study, we investigated the pharmacokinetics of naked RNA administered into the lymph node. We observed that RNA is rapidly and selectively uptaken by lymph node dendritic cells (DCs). Furthermore, in vitro and in vivo studies revealed that the efficient internalization of RNA by human and murine DCs is primarily driven by macropinocytosis. Selective inhibition of macropinocytosis by compounds or as a consequence of DC maturation abrogated RNA internalization and delivery of encoded antigens. Our findings imply that bioavailability of recombinant RNA vaccines in vivo highly depends on the density and the maturation stage of DCs at the administration site and are of importance for the design of RNA-based clinical immunotherapy protocols.

Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo.

Vaccination with in vitro transcribed RNA coding for tumor antigens is considered a promising approach for cancer immunotherapy and has already entered human clinical testing. One of the basic objectives for development of RNA as a drug is the optimization of immunobioavailability of the encoded antigen in vivo. By analyzing the effect of different synthetic 5' mRNA cap analogs on the kinetics of the encoded protein, we found that m(2)(7,2'-O)Gpp(S)pG (beta-S-ARCA) phosphorothioate caps, in particular the D1 diastereoisomer, profoundly enhance RNA stability and translational efficiency in immature but not mature dendritic cells. Moreover, in vivo delivery of the antigen as beta-S-ARCA(D1)-capped RNA species is superior for protein expression and for efficient priming and expansion of naïve antigen-specific T cells in mice. Our findings establish 5' mRNA cap analogs as yet another module for tuning immunopharmacological properties of recombinant antigen-encoding RNA for vaccination purposes.

On the specific identity of specimens of Phytoseiulus longipes Evans (Mesostigmata: Phytoseiidae) showing different feeding behaviours: morphological and molecular analyses.

This paper focuses on the differentiation of specimens, identified as Phytoseiulus longipes, collected in four countries: Argentina, Brazil, Chile and South Africa. Two of these populations are known to feed and develop on Tetranychus evansi, whereas the two others do not. As morphologically similar specimens can sometimes belong to different species and because differences in predatory behaviours exist among the four populations considered, we tested for the presence of cryptic species. Morphological and molecular experiments (12S rDNA) were carried out. The four studied populations of P. longipes could be morphologically differentiated thanks to a combination of characters. However, these morphological differences are very small. The two populations that feed and develop on T. evansi (from Argentina and Brazil) are morphologically closer to each other than to the two other populations. Genetic distances among the four populations of P. longipes were very low, suggesting that despite their different feeding habits, all specimens belong to the same species. However, the populations associated with T. evansi showed some genetic differentiation from those that do not use this pest. This is the first time that this type of differentiation has been reported for the family Phytoseiidae. These results are of primary importance to ensure the success of biological control programs and to develop strains adapted to both crops and prey species.

Kampimodromus aberrans (Acari: Phytoseiidae) from the USA: morphological and molecular assessment of its density.

Morphological measurements and a mitochondrial molecular marker (COI) were used to identity specimens reported as Kampimodromus aberrans on hazelnut in the USA. Several species and populations of this genus were studied to assist with identification. Both data types showed that specimens from the USA differed from K. aberrans from other regions. USA specimens seem to belong to the same species as Kampimodromus specimens from France on hazelnut. These mites were morphologically similar to Kampimodromus coryli and K. corylosus, which according to the original descriptions, are distinguished by the presence or absence of a tooth on the movable digit of the chelicera, with K. coryli having one tooth and K. corylosus none. As chelicerae of Kampimodromus from hazelnut in the USA and France are toothless, they are assigned to the species K. corylosus. Studies showed that morphological characters traditionally used to identify Kampimodromus species, such as setal length, are of less value than other characters that are difficult to observe, such as the numbers of solenostomes and the presence of teeth on the movable digit of the chelicerae. Some synonyms are discussed.

Cancer immunotherapy 2005: Mainz, Germany, 12-13 May 2005.

Cancer Immunotherapy 2005 was the third international meeting organized by the Association for Immunotherapy of Cancer (AIC). About 200 participants were attracted by the excellent scientific program that consisted of overview lectures from 25 international speakers in the plenary auditorium and four guided poster sessions during both days of the meeting. The first day of the symposium mainly focused on experience with, and new perspectives in, antibody therapy. On the second day of the meeting, organized as a joint conference together with the Combined Research Grant "Mechanisms of Tumor Defense and Therapeutic Intervention" funded by the German Research Council, the participants had the chance to gain deeper insights into the principles of antigen processing and the regulation of immune responses. Further topics that were discussed mainly in the poster sessions and in the special lecture given by M. Nishimura (Chicago, USA), were "cellular therapies" and "vaccination against cancer". The lectures selected for this report aim to provide an overview of the complete scientific program and give an impression of the lively atmosphere that could be felt from the first until the last session of CIMT 2005.

Failure of sustained engraftment after non-myeloablative conditioning with low-dose TBI and T cell-reduced allogeneic peripheral stem cell transplantation.

We investigated whether a T cell-reduced allogeneic stem cell transplant (SCT) with minimal conditioning and subsequent donor lymphocyte infusions (DLI) could reduce the incidence and severity of GVHD while retaining stable engraftment. Five patients with hematological malignancies (three MM, one CLL, one Chediak-Higashi syndrome) were conditioned with TBI (200 cGy). One patient additionally received fludarabine (120 mg/m(2)). CsA and mofetyl-mycophenolate (MMF) were administered to prevent GVHD. All patients were grafted with >3 x 10(6)/kg highly purified CD34(+) cells together with 2 x 10(6)/kg CD3(+) cells (three patients) or 1 x 10(5)/kg CD3(+) cells (two patients). Quick hematopoietic recovery and initial mixed donor chimerism was observed. Treatment-related toxicity was minimal in all but one patient who died of treatment-refractory GVHD on day 112. The four other patients only achieved partial donor T cell chimerism. BM and PBMC donor chimerism was lost between day 40 and 209 despite DLI. Three patients are alive with disease and one is in CR. We conclude that T cell-reduced SCT using 200 cGy as the conditioning regimen does not result in stable hematopoietic engraftment. Predominant donor T cell chimerism is not a prerequisite for initial allogeneic hematopoietic proliferation. However for sustained long-term engraftment it is of major importance.

Exposure to violence and intentions to engage in moralistic violence during early adolescence.

This study examined young adolescents'intentions to use moralistic violence and their violence exposure, examining male-female differences. Sixth-grade students (n=702) from Georgia middle schools servicing impoverished communities participated. Data were obtained on the students' exposure to violence, family structure and education level, church attendance, gang interest, drug use, and depression status. The dependent variable, intention to use moralistic violence, was measured with an 11-item scale. Linear regression models were run separately for males and females. Males had significantly higher mean intention to use moralistic violence than females (p=0.002). Males reported being exposed to violence more than females, but exposure decreased as attendance to religious services increased. For these 11-12-year-olds, unconventional peer social norms, such as witnessing violence, increased their intention to use violence while involvement in conventional activities, such as church attendance, decreased it. The protective effect was greater for males than females.

Colonization of vineyards by phytoseiid mites: their dispersal patterns in the plot and their fate.

The effect of wind and woody margins on the dispersal and population dynamics of phytoseiid mites was studied in a vine plot for a period of two years. Mites were sampled in the plot and in the surrounding vegetation (crops and natural vegetation) in order to determine phytoseiid mite abundance. The surrounding vegetation was considered to be a reservoir of phytoseiids from where the vine plot could be invaded. Directional and non-directional soil and aerial traps were placed in the plot to determine predatory mite exchange between the two areas. Colonization of the plot occurred in two stages: first, mite migration into the plot, followed by their establishment. The two-year study partially clarified the first of these two stages. Kampimodromus aberrans was the main species caught in the aerial traps. Phytoseiid mite dispersal within the vine plot seemed to be affected by both wind (direction, intensity and regularity) and phytoseiid mite density in the woody margin. However, the woody margin had a large effect only over a short distance. Some observations pointed towards an effect of other reservoir areas but it was not possible to characterize these. The population density of the phytoseiid mites in the plot increased from 1996 to 1998, but these increases are much smaller than one would expect on the basis of the number of mites migrating by air in the plot. Moreover, blocks where most mites were trapped were not the blocks where densities of phytoseiid mites on vine leaves were the largest. It therefore seems likely that not all migrants were able to develop. Their settlement pattern was not determined and this could constitute a potential research focus for the future.

Short Report: Engraftment of T-cell-depleted allogeneic haematopoietic stem cells using a reduced intensity conditioning regimen.

Graft-versus-host disease (GVHD) remains a significant complication in patients undergoing allogeneic stem cell transplantation (SCT) using a reduced intensity conditioning regimen. Although T-cell depletion (TCD) reduces the risk of GVHD after a myeloablative conditioning regimen, it is associated with an increased risk of graft failure. We have therefore examined whether TCD compromises engraftment using a fludarabine-based conditioning regimen. Fifteen patients have been transplanted using such a regimen of whom 13 underwent ex vivo TCD. All but one patient demonstrated durable engraftment and no patient receiving a TCD product developed severe GVHD. Thus, TCD may play a role in GvHD prophylaxis using such regimens.

Exposure to violence and victimization, depression, substance use, and the use of violence by young adolescents.

OBJECTIVE: To examine the relationships among exposure to violence; tobacco, alcohol, and other substance use; depression; church attendance; and the use of violence among very young adolescents. METHODS: An 86-item confidential questionnaire was administered to 722 sixth grade students (mean age = 11.9+/-0.8 years) attending 4 middle schools serving neighborhoods in and around public housing. RESULTS: Boys had a higher mean violence scale score than girls (P < or =.0001), and students living in public housing had higher violence scale scores than other students (P< or =.0001). Self-reported use of violence was significantly associated with exposure to violence (r =.45); age (r =.28); frequency of church attendance (r = -.14); depression (r =.28); the probability of being alive at age 25 (r = -.09); the frequency of use of cigarettes (r =.39), alcohol (r =.37), and multiple substances (r =.38); and interest in a gang (r =.37). When all of these variables were analyzed with multiple linear regression, multiple substance use, exposure to violence, interest in a gang, male gender, cigarette smoking, and depression level accounted for 49.7% of the variation in the use of violence scale. CONCLUSION: Recent multiple substance use and lifetime exposure to violence and victimization were the strongest correlates with the frequency that these youth reported using violence and carrying weapons.

Nutritional rickets in African American breast-fed infants.

OBJECTIVE: To analyze the characteristics of infants and children diagnosed with nutritional rickets at two medical centers in North Carolina in the 1990s. STUDY DESIGN: The physical and radiographic findings, calcium, phosphorus, alkaline phosphatase, and 25-hydroxyvitamin D levels of infants and children diagnosed with nutritional rickets at two medical centers were reviewed. Breast-feeding data were obtained from the North Carolina Women, Infants and Children Program (WIC). RESULTS: Thirty patients with nutritional rickets were first seen between 1990 and June of 1999. Over half of the cases occurred in 1998 and the first half of 1999. All patients were African American children who were breast fed without receiving supplemental vitamin D. The average duration of breast-feeding was 12.5 months. The age at diagnosis was 5 to 25 months, with a median age of 15.5 months. Growth failure was common: length was <5th percentile in 65% of cases, and weight was <5th percentile in 43%. CONCLUSION: Factors that may have contributed to the increase in referrals of children with nutritional rickets include more African American women breast-feeding, fewer infants receiving vitamin D supplements, and mothers and children exposed to less sunlight. We recommend that all dark-skinned breast-fed infants and children receive vitamin D supplementation.

Rapid identification of minor histocompatibility antigen HA-1 subtypes H and R using fluorescence-labeled oligonucleotides.

Donor-recipient disparitiy of the minor histocompatibility antigen HA-1 is relevant for the development of graft-versus-host disease after HLA-matched sibling allogeneic bone marrow transplantation in HLA-A*0201-positive individuals. Two different alleles of HA-1 with a single amino acid polymorphism have been identified. Here we describe a time- and cost-efficient method for HA-1 typing of genomic DNA, using site-specific hybridization probes with the LightCycler. This method was compared with standard techniques as sequencing or allele-specific polymerase chain reaction (PCR) and proved to be specific, reliable and reproducible. We conclude that HA-1-subtyping using fluorescent-labeled oligonucleotides represents a attractive method for the screening of samples before allogeneic transplantation in HLA-A*0201-positive individuals.

Gender differences in risk behaviors among adolescents who experience date fighting.

OBJECTIVE: Although dating violence frequently begins during adolescence, few studies have focused on date fighting in middle and high school students. Fewer studies have studied gender differences in date violence. This study examines whether gender-specific patterns of risk behaviors exist among adolescents who report date fighting. DESIGN: The study was conducted on data collected from 21 297 students in grades 8 through 12 participating in the Vermont 1995 Youth Risk Behavior Survey. Data were analyzed on 20 724 students (females = 50.1%) who reported: 1) never having been involved in a physical fight (n = 8737); 2) that their last physical fight was with a girlfriend, boyfriend, or other dating partner (n = 432); and 3) that their last fight was with someone other than a dating partner (n = 11 555). Indicators of violence (weapon carrying, being threatened, and fighting), suicide attempts, substance use, sexual behavior, and pregnancy were analyzed with chi(2) tests. Significant variables were analyzed with stepwise logistic regression. RESULTS: Of the males, 1.8% and of the females, 4.2% reported that their last fight was with a boyfriend, girlfriend, or dating partner. Risk behaviors significantly associated among females who only experienced date fighting included the number of male sexual partners in the past 3 months (adjusted odds ratio: 1. 48; 95% confidence interval: 1.26-1.74), number of suicide attempts in the past 12 months (1.55; 1.30-1.85), riding in a car with a drinking driver (1.23; 1.10-1.37), injection of illegal drugs (2.87; 1.10-7.50), use of alcohol before last sexual encounter (1.53; 1. 27-1.86), number of pregnancies (1.66; 1.26-2.21), forced sex (2.92; 2.18-3.91), and inhalant use (1.19; 1.06-1.34). Risk behaviors significantly associated among males who experienced only date fighting were sexual activity (4.11; 2.24-7.53), number of male partners in the past 3 months (1.40; 1.12-1.75), number of times of getting someone pregnant (1.68; 1.17-2.40), experiencing forced sex (2.38; 1.11-5.13), and the number of times threatened with physical violence in past 12 months (1.82; 1.53-2.17). When compared with adolescents who reported fighting with someone other than a date, risk factors significantly associated with date fighting among females were the number of male sexual partners in the past 3 months (1.21; 1.10-1.34), older age (1.21; 1.10-1.34), carrying a weapon in the past 30 days (.77;.66-.90), experiencing forced sex (1.70; 1. 30-2.22), condom non-use (1.96; 1.60-2.41), and number of times of being threatened with physical violence in past 12 months (1.11; 1. 01-1.22). The risk factors among males were the number of male sexual partners in the past 3 months (1.43; 1.28-1.60), experiencing forced sex (1.91; 1.02-3.60), and older age (1.34; 1.14-1.57). CONCLUSIONS: The patterns of risk behaviors differed among male and female adolescents reporting dating violence. Females who reported date fighting were more likely than were nonfighters to have attempted suicide, to engage in sexual and human immunodeficiency virus risk behaviors (use of injectable drugs), to have been pregnant, experienced forced sex, and to have ridden in a car with a drinking driver. Sexual behaviors, including same-gender sexual partners, forced sex, and having been threatened with physical violence, were associated with date fighting among males. These findings are important in screening adolescents at risk for date violence.date fighting, adolescence, risk behaviors, gender.

The relationship between early age of onset of initial substance use and engaging in multiple health risk behaviors among young adolescents.

BACKGROUND: Previous research based on problem-behavior theory has found that early age of onset of substance use is associated with engaging in multiple health risk behaviors among high school students. It is unknown whether these relationships begin during early adolescence. OBJECTIVE: To examine the relationships between early age of onset of cigarette, alcohol, marijuana, and cocaine use and engaging in multiple risk behaviors among middle school students. METHODS: A modified version of the Centers for Disease Control and Prevention Youth Risk Behavior Survey was administered to 2227 sixth through eighth grade students attending 53 randomly selected middle schools in North Carolina. A Health Risk Behavior Scale was constructed from 16 behaviors, including indicators of violence and weapon carrying; current substance use; nonuse of helmets when biking, in-line skating or skateboarding; not wearing a seat belt; riding with a driver who had been drinking; and suicide plans. Among this sample of middle school students, the scale had a mean (SD) of 4.1 (2.7) (range=O-15), and had a high internal reliability coefficient (alpha(=0.74). The independent variables included first time use of cigarettes, alcohol, marijuana, and cocaine at age 11 years or earlier; actual age of onset of each substance; race and ethnicity; family composition; sex; school grade; academic ranking; and older age for school grade. These data were analyzed with analysis of variance, Spearman r, and multiple linear regression. RESULTS: All the independent variables were found to be associated (P<.005) with the Health Risk Behavior Scale during the bivariate analyses. When each of these significant variables were entered into a multiple regression model, having smoked at age 11 years or younger accounted for 21.9% of the variation in the Health Risk Behavior Scale. Male sex, early marijuana or cocaine use, older age, lower academic rank, white race, and living in a 1-parent family explained an additional 19.1% of variation in the model (adjusted R2=0.41, P<.001). When the actual ages of onset of the use of substances were analyzed, in order of magnitude; age of onset of smoking; male sex; age of onset of alcohol and marijuana use; age; lower academic ranking; age of onset of cocaine use; white race; and lower academic rating accounted for 52.8% (P<.001) of the variation in the Health Risk Behavior Scale. CONCLUSION: Even when considering sociodemographic factors, early age of onset of cigarette use was the strongest correlate of the number of health risk behaviors in which these young adolescents had engaged. Early onset of use of other substances was also associated with a clustering of health risk behaviors among this sample of middle school students. The findings suggest that screening for early experimentation with tobacco and other substance use will help identify young adolescents at increased risk for engaging in multiple health risk behaviors.