Genetic analysis reveals conspecificity of two nominal species of Anaphes fairyflies (Hymenoptera: Mymaridae), egg parasitoids of Oulema leaf beetle (Coleoptera: Chrysomelidae) pests of cereal crops in Europe and of rice in East Asia.

Anaphes (Anaphes) flavipes (Foerster), a fairyfly (Hymenoptera: Mymaridae) native of Europe, is an economically important egg parasitoid for the natural control of Oulema spp. leaf beetle (Coleoptera: Chrysomelidae) pests of cereal crops such as barley, oats, rye, and wheat in Europe, and for the classical biological control of the invasive Oulema melanopus (L.) in North America. A morphologically very similar Anaphes (Anaphes) nipponicus Kuwayama, known from mainland China, Japan, Republic of Korea, Far East of Russia and Taiwan, is an egg parasitoid of Oulema oryzae (Kuwayama), a pest of rice mainly in temperate parts of East Asia. The nuclear 28S-D2 and ITS2 and the mitochondrial COI genes were used as markers to compare specimens of A. (Anaphes) flavipes reared from eggs of an Oulema sp. on barley in Germany with those of A. (Anaphes) nipponicus reared from eggs of O. oryzae on rice in Honshu Island, Japan. Because the resulting sequences are practically identical, within an expected intraspecific genetic variability, conspecificity of these two nominal species has been confirmed, and consequently A. (Anaphes) nipponicus Kuwayama, 1932, syn. n. is synonymized with A. (Anaphes) flavipes (Foerster, 1841). Taxonomic notes and illustrations are provided for the specimens of both sexes of A. (Anaphes) flavipes from Japan to facilitate their recognition.

Molecular, Morphological, and Biological Differentiation between Anagrus virlai sp. n., an Egg Parasitoid of the Corn Leafhopper Dalbulus maidis (Hemiptera: Cicadellidae) in the New World, and Anagrus incarnatus from the Palaearctic Region (Hymenoptera: Mymaridae).

The common New World egg parasitoid of the corn leafhopper Dalbulus maidis (DeLong) (Hemiptera: Cicadellidae), an economically important pest of maize from Argentina to southern USA, has long been misidentified as the Palaearctic species Anagrus incarnatus Haliday or its synonym A. breviphragma Soyka (Hymenoptera: Mymaridae). Using a combination of genetic and morphometric methods, and available biological information, specimens reared from eggs of D. maidis in Argentina and Mexico, described and illustrated here as Anagrus (Anagrus) virlai Triapitsyn sp. n., are shown to be different from those of A. incarnatus from the Palaearctic region. Mitochondrial and nuclear ribosomal DNA sequence data provide clear evidence for the separation of the two species. Anagrus virlai is also known from Brazil, Colombia, Guadeloupe (France), and Guyana.

Armored Scales and Their Parasitoids on Commercial Avocados Grown in California or Imported from Mexico.

Levels of armored scales (Hemiptera: Diaspididae) on Mexican Hass avocados imported into California over May 2008-June 2009 were monitored on 135 trucks entering the state via the Blythe border station, the entry point receiving the highest volume of fruit. Levels of live sessile scales were 3.9-fold higher than indicated in a previous survey (September 2007-April 2008) although levels of live eggs and crawlers were similar to previous levels. A survey of avocado fruit in California infested with armored scales detected four species known to be endemic but failed to find any of the seven exotic Diaspididae entering the state on Mexican fruit. Monitoring of Mexican armored scales on imported avocados from September 2007 to December 2010 recovered 10 species of parasitoids predominated by two species of Signiphora Ashmead (Hymenoptera: Signiphoridae). One of these species, Signiphora flavopalliata Ashmead, comprised 36% of all collected Mexican parasitoids and is a known hyperparasitoid. A survey of armored scale parasitoids present on commercial California avocados detected 17 genetic signatures, with only four of these in common with those detected on imported Mexican fruit. The implications of these findings are discussed.

Mathematical and computational models of drug transport in tumours.

The ability to predict how far a drug will penetrate into the tumour microenvironment within its pharmacokinetic (PK) lifespan would provide valuable information about therapeutic response. As the PK profile is directly related to the route and schedule of drug administration, an in silico tool that can predict the drug administration schedule that results in optimal drug delivery to tumours would streamline clinical trial design. This paper investigates the application of mathematical and computational modelling techniques to help improve our understanding of the fundamental mechanisms underlying drug delivery, and compares the performance of a simple model with more complex approaches. Three models of drug transport are developed, all based on the same drug binding model and parametrized by bespoke in vitro experiments. Their predictions, compared for a 'tumour cord' geometry, are qualitatively and quantitatively similar. We assess the effect of varying the PK profile of the supplied drug, and the binding affinity of the drug to tumour cells, on the concentration of drug reaching cells and the accumulated exposure of cells to drug at arbitrary distances from a supplying blood vessel. This is a contribution towards developing a useful drug transport modelling tool for informing strategies for the treatment of tumour cells which are 'pharmacokinetically resistant' to chemotherapeutic strategies.

First Report of Geosmithia pallida Causing Foamy Bark Canker, a New Disease on Coast Live Oak (Quercus agrifolia), in Association with Pseudopityophthorus pubipennis in California.

Declining coast live oak (Quercus agrifolia) trees have been observed since 2012 throughout urban landscapes in Los Angeles, Orange, Riverside, Santa Barbara, Ventura, and Monterey counties in California. Symptoms causing branch dieback and tree death included a cinnamon-colored gum seeping through multiple 0.95-mm-diameter entry holes on the bole, followed by a prolific, cream-colored foamy liquid. Beneath the outer bark was phloem and xylem necrosis. Fifty 1- to 2.5-mm adult and larval beetles were collected. Adults fit the morphological description of Pseudopityophthorus pubipennis (western oak bark beetle) (R. Rabaglia, personal communication), and ~800 bp of the mitochondrial COI gene was amplified for three beetles using primer pairs and methods previously described (2,3). All three sequences were identical (GenBank Accession Nos. KJ831289 to 91) and a BLAST search confirmed the closest match (94%) as P. pubipennis. Necrotic wood tissues collected from two trees in each county were cultured on potato dextrose agar amended with 0.01% tetracycline (PDA-tet), and incubated at 25°C for 1 week. Ochre-colored cultures with plane or radially furrowed velutinous mycelium were consistently produced. Fifty conidia each measured from two isolates were 3.66 ± 0.04 µm × 1.77 ± 0.03 µm, and arranged in non-persistent conidial chains, at first roughly parallel, becoming tangled with age. These fungal colonies were observed within gallery walls. The rDNA internal transcribed spacer (ITS) was amplified using primer pairs and methods previously described (5). Three isolates were sequenced and matched 100% to known sequences of Geosmithia pallida in GenBank; sequences of two isolates (UCR2208 and UCR2210) were deposited in GenBank (KJ468687 and KJ468688). Pathogenicity tests were performed by inoculating twelve 27.0-cm detached coast live oak shoots for each isolate with a spore suspension of G. pallida (UCR2208 and UCR2210) and sterile distilled water for controls. A 2-mm-wide, 3-mm-deep hole was drilled into the center of each shoot, 20 µl of a 106 conidia/ml spore suspension was pipetted into the hole, and sealed with Vaseline and Parafilm. The experiment was repeated twice. After 4 weeks in a moist chamber at 25°C, lesions produced by G. pallida averaged 8.3 cm and was significantly longer (ANOVA; P < 0.0001) from the control (average 0.4 cm). G. pallida was re-isolated from all inoculated plants and identified by colony morphology. P. pubipennis is a native beetle, common as a secondary agent, and previously not associated with disease. However, cryptic species may be common among bark and ambrosia beetles (4). A larger sample (i.e., populations and loci) is needed to determine the precise taxonomic status of P. pubipennis. G. pallida was shown to inhibit root growth of Q. petraea by 25% in Europe (1), appears to have affinities with a range of subcorticolous insects, and is widely distributed (5), but there is no published record of the fungus occurring in the United States. This is the first report of G. pallida causing foamy bark canker in association with P. pubipennis on Q. agrifolia in California. Results suggest this new disease complex is causing decline of Q. agrifolia throughout the state. References: (1) D. Cizková et al. Folia Microbiol. 50:59, 2005. (2) A. I. Cognato and F. A. H. Sperling. Mol. Phylogenet. Evol. 14:445, 2000. (3) A. I. Cognato et al. Mol. Phylogenet. Evol. 36:494, 2006. (4) B. H. Jordal and M. Kambestad. Mol. Ecol. Res. 14:7, 2014. (5) M. Kolarík et al. Mycol. Res. 108:1053, 2004.

Lymphangiogenesis in abdominal aortic aneurysm.

BACKGROUND: Ongoing angiogenesis is implicated in the inflammatory environment that characterizes abdominal aortic aneurysm (AAA). Although lymphangiogenesis has been associated with chronic inflammatory conditions, it has yet to be demonstrated in AAA. The aim was to determine the presence of lymphangiogenesis and to delineate the relationship between inflammation and neovascularization in AAA tissue. METHODS: AAA samples and preoperative computed tomography images were obtained from patients undergoing elective AAA repair. Control samples were age-matched abdominal aortic tissue. Specific immunostains for blood vessels (CD31, CD105), lymphatic vessels (D2-40), vascular endothelial growth factor (VEGF) A and VEGF receptor (VEGFR) 3 allowed characterization and quantitation of vasculature. RESULTS: The AAA wall contained high levels of inflammatory infiltrate; microvascular densities of blood (P < 0·001) and lymphatic (P = 0·003) vessels were significantly increased in AAA samples compared with controls. Maximal AAA vascularity was observed in inflammatory areas, with vessels that stained positively for CD31 (ρ = 0·625, P = 0·017), CD105 (ρ = 0·692, P = 0·009) and D2-40 (ρ = 0·675, P = 0·008) correlating positively with the extent of inflammation. Increased VEGFR-3 and VEGF-A expression was also evident within inflammatory AAA areas. CONCLUSION: These findings demonstrated lymphatic vessel involvement in end-stage AAA disease, which was associated with the degree of inflammation, and confirmed the involvement of neovascularization.

Phylogeographic structure, outbreeding depression, and reluctant virgin oviposition in the bean thrips, Caliothrips fasciatus (Pergande) (Thysanoptera: Thripidae), in California.

Bean thrips, Caliothrips fasciatus, is native to western North America. Once considered a pest of several crops in its native area, its pest status has waned over recent decades. However, due to its habit of aggregating in the navel of navel oranges, bean thrips remains economically important because some countries importing oranges from California have designated it a quarantine pest. Despite continued propagule pressure, bean thrips has never established outside North America. We examined genetic variation in mitochondrial DNA among Californian populations of C. fasciatus and found that potentially two cryptic species are present (supported by Kimura 2-P distances): a common widespread form B and a rarer form A with a very limited distribution. Form B showed strong phylogeographic structure, with many haplotypes having a limited geographic distribution. Inter-population crossing experiments between three geographically isolated populations of form B resulted in the production of some female offspring, indicating a degree of compatibility between these populations of this haplodiploid species. However, substantial outbreeding depression was also detected. A low frequency of offspring production by hetero-population pairs was evidence of pre-mating isolation, while post-mating isolation was also evident in the elevated mortality of fertilized eggs in successful hetero-population crosses. One surprising finding was the total lack of offspring production by virgin females when isolated individually. However, virgin females did produce sons in the presence of other virgin females. A test for the presence of Wolbachia showed that form B was not infected, but that some populations of the rarer form A were.

Genetic structure of Graphocephala atropunctata (Hemiptera: Cicadellidae) populations across its natural range in California reveals isolation by distance.

The genetic structure of 23 populations of Graphocephala atropunctata (Signoret) (Hemiptera: Cicadellidae: Cicadellinae), a vector of the plant pathogenic bacterium Xylellafastidiosa Wells et al., was investigated using ribosomal 28S and mitochondrial cytochrome C oxidase I gene sequences. The 28S sequences were identical across all G. atropunctata specimens and populations, but 16 mitochondrial haplotypes were detected and significant interpopulation differences were found in the distribution of these haplotypes. Pairwise estimates of Fst correlated positively with geographical distance between populations, a phenomenon known as genetic isolation by distance. Thus, despite potential for widespread movement of G. atropunctata through nursery and agricultural industries, isolated populations of G. atropunctata have remained genetically distinct, suggesting that negligible numbers of G. atropunctata are actually transported or population interbreeding rarely occurs. The phylogenetic relationship between G. atropunctata and two additional congeners, Graphocephala cythura Baker and Graphocephala flavovittata Metcalf, which have overlapping distributions with G. atropunctata, also was investigated. Although 28S sequences of G. flavovittata were strikingly similar to those of G. atropunctata, mitochondrial DNA (mtDNA) suggests that both species are genetically distinct from G. atropunctata.

Phoretic dispersal of armored scale crawlers (Hemiptera: Diaspididae).

Dispersal and colonization of new areas by armored scale insects (Hemiptera: Diaspididae) is achieved by mobile first-instar nymphs, called crawlers. Few studies have considered the actual mechanisms by which crawlers disperse, and although crawlers are capable of actively wandering over short distances (generally < 1 m), their dispersal over longer distances has been thought to be wind-mediated. Here, we present evidence of a potentially more important means of dispersal over longer distances (> 1 m). We first confirmed that crawlers of four species of Diaspididae [Abgrallaspis aguacatae Evans, Watson & Miller; Hemiberlesia lataniae (Signoret); Aspidiotus nerii Bouché; and Diaspidiotus perniciosus (Comstock)] have four hairs on the end of each of their legs and that each of these hairs ends in a suction cup-like structure, reminiscent of the attachment structures possessed by phoretic mites. In a controlled environment, using crawlers of A. nerii, we then showed that the crawlers use these structures to attach themselves to three different insect species [Musca domestica L., Cryptolaemus montrouzieri Mulsant and Linepithema humile (Mayr)] and can effectively be moved phoretically by these insects. Crawlers can remain attached to flying insects for considerable periods of time, suggesting that this may be an important means of dispersal for armored scale insects. The importance of phoresy for diaspidid dispersal in the field remains to be determined.

High levels of exotic armored scales on imported avocados raise concerns regarding USDA-APHIS' phytosanitary risk assessment.

Between 1914 and 2007, a quarantine protected California avocado, Persea americana Mill., groves from pests that might be introduced into the state along with fresh, imported avocados. Soon after Mexican avocados were first allowed entry on 1 February 2007, live specimens of several species of armored scales (Hemiptera: Diaspididae) not believed to be present in California were detected on 'Hass' avocados entering the state from Mexico. Initially, the California Department of Food and Agriculture (CDFA) prevented avocados infested with these scales from entering the state or required that they be fumigated with an approved treatment such as methyl bromide. After a Science Advisory Panel meeting in May 2007, U.S. Department of Agriculture-Animal and Plant Health Inspection Service (USDA-APHIS) reaffirmed its position that armored scales on shipments of fruit for consumption (including avocados) pose a "low risk" for pest establishment. In compliance with APHIS protocols, as of 18 July 2007, CDFA altered its policy to allow shipments of scale-infested avocados into the state without treatment. Here, we report on sampling Mexican avocados over an 8-mo period, September 2007-April 2008. An estimated 67 million Mexican Hass avocados entered California over this period. Based on samples from 140 trucks containing approximately 15.6% of this volume of fruit, we estimate that approximately 47.6 million live, sessile armored scales and an additional 20.1 million live eggs and crawlers were imported. We found eight probable species of armored scales in the samples, seven of these are not believed to occur in California; 89.3% of the live scales were Abgrallaspis aguacatae Evans, Watson and Miller, a recently described species. In contrast to the USDA-APHIS opinion, we believe the volume of shipments and levels of live scales they contain present a significant risk to California's US$300 million avocado industry and to other crops that might become infested by one or more of these exotic species.

A mathematical model of doxorubicin penetration through multicellular layers.

Inadequate drug delivery to tumours is now recognised as a key factor that limits the efficacy of anticancer drugs. Extravasation and penetration of therapeutic agents through avascular tissue are critically important processes if sufficient drug is to be delivered to be therapeutic. The purpose of this study is to develop an in silico model that will simulate the transport of the clinically used cytotoxic drug doxorubicin across multicell layers (MCLs) in vitro. Three cell lines were employed: DLD1 (human colon carcinoma), MCF7 (human breast carcinoma) and NCI/ADR-Res (doxorubicin resistant and P-glycoprotein [Pgp] overexpressing ovarian cell line). Cells were cultured on transwell culture inserts to various thicknesses and doxorubicin at various concentrations (100 or 50 microM) was added to the top chamber. The concentration of drug appearing in the bottom chamber was determined as a function of time by HPLC-MS/MS. The rate of drug penetration was inversely proportional to the thickness of the MCL. The rate and extent of doxorubicin penetration was no different in the presence of NCI/ADR-Res cells expressing Pgp compared to MCF7 cells. A mathematical model based upon the premise that the transport of doxorubicin across cell membrane bilayers occurs by a passive "flip-flop" mechanism of the drug between two membrane leaflets was constructed. The mathematical model treats the transwell apparatus as a series of compartments and the MCL is treated as a series of cell layers, separated by small intercellular spaces. This model demonstrates good agreement between predicted and actual drug penetration in vitro and may be applied to the prediction of drug transport in vivo, potentially becoming a useful tool in the study of optimal chemotherapy regimes.

Angiogenesis in breast cancer: how should we measure this? (Review).

Despite the established importance of angiogenesis in the pathogenesis of solid tumours, there is still no consensus on how this is best measured or which method is the most appropriate in the determination of prognosis. Here we review the pros and cons of current methods of assessing angiogenesis, both clinical and in the laboratory, and discuss with respect to breast cancer.

A mathematical model of tumour angiogenesis, regulated by vascular endothelial growth factor and the angiopoietins.

Angiogenesis--the growth of new blood vessels from existing ones--is a prerequisite for the growth of solid tumours beyond a diameter of approximately 2 mm. In recent years, the angiopoietins have emerged as important regulators of angiogenesis. They mediate a delicate balance between vascular quiescence, regression and new growth, but their mechanism of action is not fully understood. This work attempts to provide a mathematical description of the role of the angiopoietins in angiogenesis. The model is formulated within the framework of reinforced random walks, which allows easy transition between the continuum (macroscopic) and discrete (microscopic) forms. Model predictions are in qualitative agreement with experimental observations, and may have implications for anti-cancer therapies based on the prevention of angiogenesis.

The role of the angiopoietins in tumour angiogenesis.

Angiogenesis--the growth of new blood vessels from existing ones--is a prerequisite for the growth of solid tumours beyond a diameter of approximately 2 mm. In recent years, the angiopoietins have emerged as important regulators of angiogenesis. They mediate a delicate balance between vascular quiescence, regression and new growth, but their mechanism of action is not fully understood. This work attempts to provide a mathematical description of the role of the angiopoietins in angiogenesis. The model is formulated within the framework of reinforced random walks, which allows easy transition between the continuum (macroscopic) and discrete (microscopic) forms. Model predictions are in qualitative agreement with experimental observations, and may have implications for antiangiogenic cancer therapies.

Eight new HLA-A alleles associated with antigens in the A2 CREG.

This report describes eight novel HLA-A alleles. Seven of these new alleles contain substitutions previously reported in other HLA-A alleles, suggesting an origin resulting from a gene conversion mechanism. Of these seven, A*2431 contains a substitution previously associated with a larger polymorphic motif. A*0247 contains a substitution at a previously polymorphic position, but encodes an amino acid change previously unreported in any HLA-A, -B or -C alleles.

Suspected acute appendicitis: trends in management over 30 years.

BACKGROUND: Tradition taught that patients with signs suspicious of acute appendicitis should be explored surgically, but studies in the 1960s found that if this group was closely observed about one-third recovered without treatment. To differentiate these patients a strict regimen of active observation was introduced and this has now been studied over 30 years. Ultrasonography, laparoscopy and computed tomography (CT) have also been used to clarify the diagnosis in these patients. METHODS: Papers on the management of patients with suspected appendicitis published since 1970 were traced through Index Medicus, English-language journals and Medline. All those that mention the use of observation are reviewed, with selective reports on the other methods used. The advantages and disadvantages of various methods of management are compared. RESULTS AND CONCLUSION: Active observation has yielded a consistently low negative appendicectomy rate without a rise in the perforation rate; there was one death in over 1600 patients. It has proved to be a practical method of discriminating between patients who do and do not need an operation. In this situation, both ultrasonography and CT yield some false-negative reports, so results must be checked at the bedside. Furthermore, CT involves exposure to significant doses of radiation.

Twenty-nine new HLA-B alleles associated with antigens in the 5C CREG.

This paper describes 29 novel HLA-B locus alleles identified during low-resolution typing. The majority of the novel alleles carry new patterns of previously known polymorphic motifs or codons. Three alleles carry alterations in the Bw4/Bw6 epitope. Five alleles carry novel substitutions.

Novel HLA-B alleles associated with antigens in the 8C CREG.

This paper describes 13 novel HLA-B locus alleles, B*0809, B*0812, B*0813, B*0814, B*14062, B*3804, B*3806, B*3914, B*3915, B*3918, B*3919, B*3920, and B*3922 which represent new patterns of known polymorphic residues.