Three-Dimensional Trapping of Individual Rydberg Atoms in Ponderomotive Bottle Beam Traps.

We demonstrate three-dimensional trapping of individual Rydberg atoms in holographic optical bottle beam traps. Starting with cold, ground-state ^{87}Rb atoms held in standard optical tweezers, we excite them to nS_{1/2}, nP_{1/2}, or nD_{3/2} Rydberg states and transfer them to a hollow trap at 850 nm. For principal quantum numbers 60≤n≤90, the measured trapping time coincides with the Rydberg state lifetime in a 300 K environment. We show that these traps are compatible with quantum information and simulation tasks by performing single qubit microwave Rabi flopping, as well as by measuring the interaction-induced, coherent spin-exchange dynamics between two trapped Rydberg atoms separated by 40 µm. These results will find applications in the realization of high-fidelity quantum simulations and quantum logic operations with Rydberg atoms.

L-System model for the growth of arbuscular mycorrhizal fungi, both within and outside of their host roots.

Development of arbuscular mycorrhizal fungal colonization of roots and the surrounding soil is the central process of mycorrhizal symbiosis, important for ecosystem functioning and commercial inoculum applications. To improve mechanistic understanding of this highly spatially and temporarily dynamic process, we developed a three-dimensional model taking into account growth of the roots and hyphae. It is for the first time that infection within the root system is simulated dynamically and in a spatially resolved way. Comparison between data measured in a calibration experiment and simulated results showed a good fit. Our simulations showed that the position of the fungal inoculum affects the sensitivity of hyphal growth parameters. Variation in speed of secondary infection and hyphal lifetime had a different effect on root infection and hyphal length, respectively, depending on whether the inoculum was concentrated or dispersed. For other parameters (branching rate, distance between entry points), the relative effect was the same independent of inoculum placement. The model also indicated that maximum root colonization levels well below 100%, often observed experimentally, may be a result of differential spread of roots and hyphae, besides intrinsic plant control, particularly upon localized placement of inoculum and slow secondary infection.

Root induced changes of effective 1D hydraulic properties in a soil column.

AIMS: Roots are essential drivers of soil structure and pore formation. This study aimed at quantifying root induced changes of the pore size distribution (PSD). The focus was on the extent of clogging vs. formation of pores during active root growth. METHODS: Parameters of Kosugi's lognormal PSD model were determined by inverse estimation in a column experiment with two cover crops (mustard, rye) and an unplanted control. Pore dynamics were described using a convection-dispersion like pore evolution model. RESULTS: Rooted treatments showed a wider range of pore radii with increasing volumes of large macropores >500 µm and micropores <2.5 µm, while fine macropores, mesopores and larger micropores decreased. The non-rooted control showed narrowing of the PSD and reduced porosity over all radius classes. The pore evolution model accurately described root induced changes, while structure degradation in the non-rooted control was not captured properly. Our study demonstrated significant short term root effects with heterogenization of the pore system as dominant process of root induced structure formation. CONCLUSIONS: Pore clogging is suggested as a partial cause for reduced pore volume. The important change in micro- and large macropores however indicates that multiple mechanic and biochemical processes are involved in root-pore interactions.

Selected reaction monitoring method to determine the species origin of blood-based binding agents in meats: a collaborative study.

Binding products or food 'glues' are used throughout the food industry to increase the meat use rate or to augment economic efficiency. Some of these binders contain thrombin from bovine and porcine blood. The European parliament has recently banned thrombin-based additives and labelling legislation governs their use in the US. A mass spectrometry screening method is available to detect the addition of thrombin agents to foods as there is a need to protect consumers and to avoid misleading trade practices. We report the details of an inter-laboratory trial to determine the transferability of this method to operators in various food testing laboratories, each using a different triple quadrupole mass spectrometer design. The trial was successful with the species origin of the binding agent contained in each of the 43 test materials being correctly reported by the participants. This is consistent with a false positive and false negative rate of 0%. This is the first collaborative study, as far as we are aware, which involves a liquid chromatography mass spectrometry (LC-MS/MS) application to approach a food authenticity issue.

Field quantification of wetting-drying cycles to predict temporal changes of soil pore size distribution.

Wetting-drying (WD) cycles substantially influence structure related soil properties and processes. Most studies on WD effects are based on controlled cycles under laboratory conditions. Our objective was the quantification of WD cycles from field water content measurements and the analysis of their relation to the temporal drift in the soil pore size distribution. Parameters of the Kosugi hydraulic property model (rm,Kosugi, σKosugi) were derived by inverse optimization from tension infiltrometer measurements. Spectral analysis was used to calculate WD cycle intensity, number and duration from water content time series. WD cycle intensity was the best predictor (r2 = 0.53-0.57) for the temporal drift in median pore radius (rm,Kosugi) and pore radius standard deviation (σKosugi). At lower soil moisture conditions the effect of cycle intensity was reduced. A bivariate regression model was derived with WD intensity and a meteorological indicator for drying periods (ET0, climatic water balance deficit) as predictor variables. This model showed that WD enhanced macroporosity (higher rm,Kosugi) while decreasing pore heterogeneity (lower σKosugi). A drying period with high cumulative values of ET0 or a strong climatic water balance deficit on the contrary reduced rm,Kosugi while slightly increasing σKosugi due to higher frequency at small pore radius classes. The two parameter regression model was applied to predict the time course of soil pore size distribution parameters. The observed system dynamics was captured substantially better by the calculated values compared to a static representation with averaged hydraulic parameters. The study showed that spectral analysis is an adequate approach for the quantification of field WD pattern and that WD intensity is a key factor for the temporal dynamics of the soil pore size distribution.

Environmental and management influences on temporal variability of near saturated soil hydraulic properties.

Structural porosity is a decisive property for soil productivity and soil environmental functions. Hydraulic properties in the structural range vary over time in response to management and environmental influences. Although this is widely recognized, there are few field studies that determine dominant driving forces underlying hydraulic property dynamics. During a three year field experiment we measured temporal variability of soil hydraulic properties by tension infiltrometry. Soil properties were characterized by hydraulic conductivity, effective macroporosity and Kosugi's lognormal pore size distribution model. Management related influences comprised three soil cover treatment (mustard and rye vs. fallow) and an initial mechanical soil disturbance with a rotary harrow. Environmental driving forces were derived from meteorological and soil moisture data. Soil hydraulic parameters varied over time by around one order of magnitude. The coefficient of variation of soil hydraulic conductivity K(h) decreased from 69.5% at saturation to 42.1% in the more unsaturated range (- 10 cm pressure head). A slight increase in the Kosugi parameter showing pore heterogeneity was observed under the rye cover crop, reflecting an enhanced structural porosity. The other hydraulic parameters were not significantly influenced by the soil cover treatments. Seedbed preparation with a rotary harrow resulted in a fourfold increase in macroporosity and hydraulic conductivity next to saturation, and homogenized the pore radius distribution. Re-consolidation after mechanical loosening lasted over 18 months until the soil returned to its initial state. The post-tillage trend of soil settlement could be approximated by an exponential decay function. Among environmental factors, wetting-drying cycles were identified as dominant driving force explaining short term hydraulic property changes within the season (r2 = 0.43 to 0.59). Our results suggested that beside considering average management induced changes in soil properties (e.g. cover crop introduction), a dynamic approach to hydrological modeling is required to capture over-seasonal (tillage driven) and short term (environmental driven) variability in hydraulic parameters.

Maternal serum proteome changes between the first and third trimester of pregnancy in rural southern Nepal.

Characterization of normal changes in the serum proteome during pregnancy may enhance understanding of maternal physiology and lead to the development of new gestational biomarkers. In 23 Nepalese pregnant women who delivered at term, two-dimensional difference in-gel electrophoresis (DIGE) was used to assess changes in relative protein abundance between paired serum samples collected in the first and third trimesters. One-hundred and forty-five of over 700 protein spots in DIGE gels (pI 4.2-6.8) exhibited nominally significant (p < 0.05) differences in abundance across trimesters. Additional filtering using a Bonferroni correction reduced the number of significant (p < 0.00019) spots to 61. Mass spectrometric analysis detected 38 proteins associated with gestational age, cytoskeletal remodeling, blood pressure regulation, lipid and nutrient transport, and inflammation. One new protein, pregnancy-specific ß-glycoprotein 4 was detected. A follow-up isotope tagging for relative and absolute quantitation (iTRAQ) experiment of six mothers from the DIGE study revealed 111 proteins, of which 11 exhibited significant (p < 0.05) differences between trimesters. Four of these proteins: gelsolin, complement C1r subcomponent, α-1-acid glycoprotein, and α-1B-glycoprotein also changed in the DIGE analysis. Although not previously associated with normal pregnancy, gelsolin decreased in abundance by the third trimester (p < 0.01) in DIGE, iTRAQ and Western analyses. Changes in abundance of proteins in serum that are associated with syncytiotrophoblasts (gelsolin, pregnancy-specific ß-1 glycoprotein 1 and ß-2-glycoprotein I) probably reflect dynamics of a placental proteome shed into maternal circulation during pregnancy. Measurement of changes in the maternal serum proteome, when linked with birth outcomes, may yield biomarkers for tracking reproductive health in resource poor settings in future studies.

Dispersal of stable flies (Diptera: Muscidae) from larval development sites in a Nebraska landscape.

Seven mark-recapture studies were conducted over 3 yr to assess dispersal of newly emerging adult stable flies, Stomoxys calcitrans L., from larval development sites in a mixed agricultural environment in northeastern Nebraska. Infested hay debris piles were marked by dusting their surfaces with fluorescent pigments, adults were captured with surrounding grids of Alsynite sticky traps, and specimens were dissected to determine feeding histories and reproductive age. Distances and directions of 3,889 marked specimens indicated males and females dispersed equally and in all directions. Midguts of males and females were equally likely to contain blood-meal remnants. Percentage with blood remnants and percentage of females with yolk increased with distance from mark origin, indicating survival and spread were positively associated with host finding success. A time-integrated diffusion model fit to results from the seven studies indicated 50% of stable fly adults had dispersed beyond 1.6 km of their natal site, but only 5% had dispersed beyond 5.1 km. These results indicate that stable fly adults on cattle in a given area are most likely to have originated from larval development sites within an ≈ 5 km radius of the subject cattle.

Bioavailable iron and heme metabolism in Plasmodium falciparum.

Iron metabolism is essential for cell function and potentially toxic because iron can catalyze oxygen radical production. Malaria-attributable anemia and iron deficiency anemia coincide as being treatable diseases in the developing world. In absolute amounts, more than 95% of Plasmodium metal biochemistry occurs in the acidic digestive vacuole where heme released from hemoglobin catabolism forms heme crystals. The antimalarial quinolines interfere with crystallization. Despite the completion of the Plasmodium genome, many 'gene gaps' exist in components of the metal pathways described in mammalian or yeast cells. Present evidence suggests that parasite bioavailable iron originates from a labile erythrocyte cytosolic pool rather than from abundant heme iron. Indeed the parasite has to make its own heme within two separate organelles, the mitochondrion and the apicomplast. Paradoxically, despite the abundance of iron within the erythrocyte, iron chelators are cytocidal to the Plasmodium parasite. Hemozoin has become a sensitive biomarker for laser desorption mass spectrometry detection of Plasmodium infection in both mice and humans.

Management of myiasis: current status and future prospects.

The management of myiasis in livestock has been an example of the success of modern chemical approaches for parasite control, yet in some cases remains extremely intractable, requiring the development of novel strategies. In addition, the growing and urgent need to develop integrated strategies that enhance the sustainability of livestock production systems drives the search for new techniques [see Int. J. Parasitol. 29 (1999) 7].The following summary represents a synthesis of a symposium presented at the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology, New Orleans,USA, 10­14 August 2003. The coverage began with a review of the need for more subtle economic analysis of the impact of myiasis based on the use of the sterile insect technique (SIT) for control of bovine hypodermosis in North America. This was followed by a review of the status of chemical control with particular emphasis on the macrocyclic lactones. The outcome of the use of these compounds in a regulated control program for eradication of bovine hypodermosis in EU was surveyed. Similarly, the success of the screwworm eradication program, using the sterile insect technique has shown how effective this approach can be given the appropriate target. Several aspects of the development of newer approaches were surveyed in discussion of newer chemical control products, development of vaccines, use of host genetics, use of predictive simulation modelling and trapping for monitoring and control and the development of new diagnostic approaches for occult infestations. Finally, use of the latest molecular tools for identification of larvae causing myiasis and their use for the identification of species coming from different and distant geographical areas to colonize regions where they have been eradicated was reviewed.

Detection of malaria parasites in blood by laser desorption mass spectrometry.

A novel method for the in vitro detection of the protozoan Plasmodium, the causative agent of malaria, has been developed. It comprises a protocol for cleanup of whole blood samples, followed by direct ultraviolet laser desorption (LD) time-of-flight mass spectrometry. Intense ion signals are observed from intact ferriprotoporphyrin IX (heme), sequestered by malaria parasites during their growth in human red blood cells. The LD mass spectrum of the heme is structure-specific, and the signal intensities are correlated with the sample parasitemia (number of parasites per unit volume of blood). Parasitemia levels on the order of 10 parasites/microL blood can be unambiguously detected by this method. Consideration of laser beam parameters (spot size, rastering across the sample surface) and actual sample consumption suggests that the detection limits can be further improved by at least an order of magnitude. The influence of experimental factors, such as desorbed ion polarity, laser exposure and fluence, sample size, and parasite growth stage, on the threshold for parasite detection is also addressed.

Testicular maturation in the sheep bot fly Oestrus ovis.

The process of testicular maturation in relation to intrapuparial development was studied in the sheep nasal bot fly, Oestrus ovis L. (Diptera: Oestridae). After formation of the puparium during larval-pupal apolysis and the cryptocephalic pupal stage (approximately 24-72 h), spermatogonia had undergone mitotic divisions and sperm cysts had been formed. Five days after pupariation, spermatogonia transformed into primary spermatocytes during the phanerocephalic pupal stage, and secondary spermatocytes first appeared during the pupal-adult apolysis. Secondary spermatocytes began undergoing the second meiotic division by day 8 (transparent-eye pharate adult stage). By days 9 and 10, round spermatids were present and began to elongate by day 11. By day 12, the first bundles of tailed spermatozoa had appeared. By day 15 (the yellow-orange eye pharate adult stage), round, elongated, tailed and bundled spermatids were predominant and by day 17 differentiating spermatids occupied nearly 35% of the testicular cavity, and 60% was occupied by free sperm. By day 21 (the red-brown eye pharate adult stage), spermatozoa colonized the seminal vesicle. At emergence (approximately day 22), a complement of free sperm occupied the testis and the seminal vesicle, but groups of developing cells frequently remained in certain zones. Spermatogenesis was carried out after pupariation and spermiogenesis occurred during the pharate adult stage. After emergence, males possessed fully formed spermatozoa ready for ejaculation.

Liquid chromatography electrospray-mass spectrometry of urinary aflatoxin biomarkers: characterization and application to dosimetry and chemoprevention in rats.

A liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS/MS) method for the measurement of aflatoxin biomarkers in urine has been developed and validated. The two major aflatoxin-DNA adducts formed in rat tissues, aflatoxin N(7)-guanine and its imidazole ring opened derivative, 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-aflatoxin B(1), were detected and quantified in urine by the LC-ESI-MS/MS technique. Other metabolites derived from the conjugation and/or oxidation of aflatoxin B(1) measured in the urine of dosed rats included aflatoxin P(1), aflatoxin P(1)-glucuronide, aflatoxin Q(1), aflatoxin M(1), 8,9-dihydro-8,9-dihydroxy aflatoxin B(1), aflatoxin B(1)-mercapturic acid, the aflatoxin-cysteine glycine adduct derived from the aflatoxin-glutathione conjugate, aflatoxin M(1)P(1) and the aflatoxin B(1)-dialcohol. For in vivo studies to determine the dosimetry of certain aflatoxin metabolites, aflatoxin B(2) was used as an internal standard for recovery since this compound is not naturally produced in rats. In the final method using the internal standard, the coefficient of variation of six replicate analyses of in vivo rat urine samples for aflatoxin N(7)-guanine, aflatoxin B(1)-mercapturic acid, and aflatoxin M(1) was 12.5, 12.8, and 5.8%, respectively. Further, the LC-ESI-MS/MS method to detect aflatoxin N(7)-guanine in in vivo rat urine samples was at least 20-fold more sensitive than prior techniques. Using the LC-ESI-MS/MS technique, the dosimetry, on a weekly basis, of major urinary aflatoxin metabolites was assessed in animals chronically dosed over a 5-week period. Of particular importance was the application of this method to determine the modulation of levels of urinary aflatoxin metabolites by treatment with oltipraz, a chemopreventive agent that can completely ablate aflatoxin hepatocarcinogenesis in the rat. After 1 week, oltipraz administration diminished urinary aflatoxin N(7)-guanine, aflatoxin B(1)-mercapturic acid and aflatoxin M(1) levels by 83, 92, and 82%, respectively. The magnitude of this reduction was persistent at the day 14, 21, 28, and 35-day time points with the average decrease of aflatoxin N(7)-guanine, aflatoxin B(1)-mercapturic acid and aflatoxin M(1) being 73, 92, and 90%, respectively. Importantly, even under circumstances where the oltipraz intervention was most efficient in reducing aflatoxin metabolite levels, the LC-ESI-MS/MS method was still sensitive enough to detect the reduced biomarker content. This outcome has important translational implications for the application and analysis of the efficacy of primary and secondary prevention interventions in human populations where ambient exposure levels are low, but the toxicologic hazards of these exposures remain high.

Coexpression of normal and mutated CD40 ligand with deletion of a putative RNA lariat branchpoint sequence in X-linked hyper-IgM syndrome.

We describe a novel CD40 ligand (CD40L) splicing mutation in a patient with X-linked hyper-IgM syndrome (X-HIM) associated with alternate splicing of exon 3, resulting in the expression of both full-length and exon-3-skipped CD40L mRNA. The mutation is an 8-bp deletion 25 bp upstream of the intron 2/exon 3 junction which overlaps a putative RNA branchpoint, suggesting that it may impair RNA lariat formation. The exon-3-skipped CD40L transcript encodes a truncated protein (CD40LDeltaE3) encompassing the cytoplasmic, transmembrane, and extracellular stalk domains, but lacking the CD40L receptor binding domain. CD40LDeltaE3 protein expression was readily detectable in transfected Cos cells by immunofluorescence. In cells cotransfected with CD40LDeltaE3 and wild-type CD40L, expression of CD40LDeltaE3 did not inhibit the expression of wild-type CD40L monomers, but strongly inhibited staining by the conformationally sensitive anti-CD40L mAb 5c8, suggesting that CD40LDeltaE3 acts in a dominant negative manner to inhibit the assembly of functional CD40L trimers. This mechanism may contribute to the pathophysiology of CD40L deficiency in X-HIM patients with leaky splice site mutations.

Oestrus ovis in sheep: relative third-instar populations, risks of infection and parasitic control.

Oestrus ovis (L.) (Diptera: Oestridae), the nasal bot fly, has a relatively short free-living life cycle outside of the host, and therefore it is necessary to know when the parasitic period occurs in order to prevent the clinical signs and economic losses caused by this parasite. The length of this parasitic portion of the life cycle is quite variable: a few weeks to several months depending on the season and climatic conditions. Surveys of Oestrus ovis larval populations in sheep show different results on the number of generations according to the local climate. Mean monthly larval profiles of L1 and L3 burdens of sheep from West African Sahelian countries, Mediterranean countries (Morocco, Tunisia and Sicily) and Southwest France were compared. Valuable information on the suspected extension of the fly season is obtained showing the period of infection in each area. This knowledge will be a valuable tool to help in choosing the right treatment at the right period.

Periodic fever syndromes.

The term periodic fever syndrome has been used in a restricted sense to denote two diseases in which episodic fevers occur with a regular periodicity: cyclic neutropenia and the periodic fever, aphthous stomatitis, pharyngitis, and adenopathy (PFAPA) syndrome. Other authors have used the term in a more general sense to encompass a larger group of disorders characterized by recurrent episodes of fever that do not necessarily follow a strictly periodic pattern. These include familial Mediterranean fever, the autosomal dominant familial fevers (also known as Hibernian fever), and the hyperimmunoglobulin D syndrome. This article follows the latter usage, and reviews recent advances in our understanding of the genetics and molecular pathology of this group of diseases, as well as their clinical characterization and treatment.

Pyrimido[1,2-alpha]purin-10(3H)-one: a reactive electrophile in the genome.

Malondialdehyde and base propenal react with deoxyguanosine residues in DNA to form an exocyclic adduct, pyrimido[1, 2-alpha]purin-10(3H)-one (1), that has been detected at high levels in genomic DNA of healthy humans. Previous studies have shown that tris(hydroxymethyl)aminomethane adds to 1 at elevated pH, forming an enaminoimine (2), but it is uncertain whether 1 reacts directly or hydrolyzes under basic conditions to N(2)-(3-oxo-1-propenyl)deoxyguanosine (3) prior to amine addition. We report that 1 reacts at neutral pH with hydroxylamines to form oximes. The rate of reaction of 1 with hydroxylamines at pH 7 is at least 150 times faster than the rate of hydrolysis of 1 to 3. Thus, 1 is directly reactive to nucleophiles. These observations indicate that 1 is an electrophile in the human genome that may react with cellular nucleophiles to form novel cross-linked adducts.

Factors affecting the larvipositional activity of Oestrus ovis gravid females (Diptera: oestridae).

Observations of fly strikes or larvipositions (n=68 in 21 days of observation) were carried out in a herd of goats during the spring in Baja California Sur, Mexico in order to identify the climatic conditions favoring larviposition activity of gravid Oestrus ovis L. flies, as well as to investigate whether a mixture of some potentially useful compounds was involved in this behavior. Hand-caught, tethered flies (n=43) were either exposed or unexposed to a combination of carbon dioxide, humidity, 1-octen-3-ol, butyric, propionic, acetic acid and acetone released from movable sheep and goat dummies under open field and cage conditions. Fly strikes occurred at temperatures greater than 20 degrees C, but mainly between 25 and 28 degrees C and from 116 to 838W m(-2) of solar irradiance. Few or no strikes were seen under moderate or strong wind, but did occur in a wide range of relative humidity. The chemicals applied did not improve the capacity of animal dummies to induce the flies to larviposit, but very irregular behavior was observed. Fourteen larvipositions were made on the dummies lacking chemical stimuli, so visual ability and movement by the dummies was very important in stimulation of the flies. Temperature appeared to be the main factor determining fly activity, but wind and solar irradiance also played important roles. Characteristics of O. ovis larviposition are discussed.

Mass spectrometry for genotyping: an emerging tool for molecular medicine.

Recent technological innovations have made proteins and nucleic acids accessible to mass spectrometric analysis. As a result of their inherently high specificity, accuracy and throughput, there is considerable interest in developing mass spectrometric methods for genotype analysis in clinical diagnostic and research applications. This review outlines some of the most promising genotyping methods developed using electrospray and matrix-assisted laser-desorption-ionization mass spectrometry.

Experimental bovine hypodermosis in Spain.

An experimental survey was carried out in western Spain to investigate both the chronobiology of Hypoderma spp. and the immunoresponse of their bovine hosts. This study was initiated with a new system of obtaining Hypoderma spp. larvae directly from their host, including the eclosion of adults from their pupae, infestation under natural but controlled conditions, and confirmation of the resulting infection. This survey was carried out over 2 cattle grub seasons; it was possible to infest and reinfest the experimental animals and to monitor them by both parasitological methods and by an enzyme-linked immunosorbent assay. This method permitted the evaluation of the development of anti-Hypoderma antibodies during the experiment. The experimental design also enabled us to establish the period of detectable H. lineatum infection to be from December until the end of April with the largest number of warbles observed during March and April. After a pupal period of < 30 d, adults were seen in April and May. Hypoderma bovis (de Geer) showed a delay of 2 m.o. relative to H. lineatum (de Villiers). This study reports a completed biological life cycle of Hypoderma spp. under controlled conditions in both natural and experimental environments.