Assessment of potential (inhalation and dermal) and actual exposure to acetamiprid by greenhouse applicators using liquid chromatography-tandem mass spectrometry

New analytical methods based on liquid chromatography with electrospray tandem mass spectrometry (LC-MS/MS) have been developed and validated for assessing the exposure of greenhouse workers to acetamiprid. Both ambient (potential inhalation and dermal exposure) and internal dose (biological monitoring of urine samples) measurements were carried out. Potential inhalation exposure was assessed using Chromosorb 102 cartridges connected to air personal samplers. Potential dermal exposure was estimated by using whole body dosimetry. The measurement of actual exposure was done by analyzing the parent compound in urine samples of the applicators, after a solid-phase extraction (SPE) step. The methods showed a good accuracy (72-92%), precision (2-13%) and lower limits (few microg l(-1)). The validated approaches have been applied to assess potential and actual exposure of agricultural workers spraying acetamiprid in greenhouses. The results shown the need to wear personal protective equipment (suits) in order to reduce the absorbed dose of acetamiprid.     

Antibody dynamics in childhood diseases: waning and boosting of immunity and the impact of vaccination

The introduction of vaccination against acute diseases such as measles induced a dramatic decline in the prevalence of the disease, and a more gradual rise in the proportion of the population whose immunity is derived solely from vaccination. These two factors combine to constitute an important shift in the dynamics of immunity, especially in highly vaccinated populations. We develop a general model to describe both loss of immunity in the absence of disease, and boosting of immunity corresponding to subclinical infection in individuals whose immunity has waned. We consider the interaction between infection and immunity and identify the key parameters that determine the eradication threshold. We explore the dynamics in the years following the introduction of vaccination using a stochastic version of the model, and consider the effect of different assumptions concerning the nature of immunity. A comparison of the model results with recently published data suggests that heterogeneity in host immune response is an important feature of the antibody dynamics.     

The protozoan parasite, Theileria annulata, induces a distinct acute phase protein response in cattle that is associated with pathology

Acute phase proteins (APP) are synthesised in the liver in response to the systemic presence of high levels of pro-inflammatory cytokines. Bacteria are considered to be strong inducers of APP whereas viruses are weak or non-inducers of APP. Very few reports have been published on APP induction by parasites. Here, we report that the tick-borne protozoan parasite of cattle, Theileria annulata, induced an atypical acute phase response in cattle. Following experimental infection, serum amyloid A (SAA) appeared first, followed by a rise in alpha(1) acid glycoprotein (alpha(1)AGP) in all animals, whereas haptoglobin, which is a major APP in cattle, only appeared in some of the animals, and generally at a low level. All three APP only became elevated around or after the appearance of schizonts in draining lymph nodes and after the first observed temperature rise. Increased alpha(1)AGP levels coincided with the appearance of piroplasms. The production of SAA and alpha(1)AGP correlated strongly with each other, and also with some clinical measures of disease severity including the time to fever, development of leucopaenia, parasitaemia and mortality. These results are consistent with the hypothesis that T. annulata causes severe pathology in susceptible cattle by inducing high levels of pro-inflammatory cytokines.     

Helminth community structure in an expanding white-winged dove (Zenaida asiatica asiatica) population

Helminth communities of 171 fledged white-winged doves (Zenaida asiatica asiatica) from the expanding eastern population in Texas (USA) were examined from hosts collected 11 June to 19 September 1997 within their historical range, new breeding periphery, and an intermediate area. Eleven helminth species, representing 435 individuals, were found. Helminths occurred in three microhabitats, of which the small intestine was the most commonly occupied. Nematodes dominated numerically (76% of total worms), followed by cestodes (17%), and trematodes (7%). Infracommunities were species-poor; the most complex infracommunity consisted of three helminth species, which occurred in three host individuals, followed by two species that occurred in 13 hosts. The remaining 155 doves had one (70) or no (85) species. The overall helminth component community was species-poor and was dominated by Ascaridia columbae which occurred in 26% of the white-winged doves and accounted for 65% of all helminth individuals. Prevalence and abundance of A. columbae varied by geographic region and host age, but not by host sex. Helminth component communities varied by geographic region, host age, and host sex. These differences were primarily attributable to unique occurrences of uncommon species within specific host subpopulations. Results suggest that the white-winged doves' multimodal regional abundance pattern, sympatry with other columbids, and granivorous diet may be more important in shaping helminth community structure than the influences often associated with geographic range expansion.     

An induced Ets repressor complex regulates growth arrest during terminal macrophage differentiation

Defining the molecular mechanisms that coordinately regulate proliferation and differentiation is a central issue in development. Here, we describe a mechanism in which induction of the Ets repressor METS/PE1 links terminal differentiation to cell cycle arrest. Using macrophages as a model, we provide evidence that METS/PE1 blocks Ras-dependent proliferation without inhibiting Ras-dependent expression of cell type-specific genes by selectively replacing Ets activators on the promoters of cell cycle control genes. Antiproliferative effects of METS require its interaction with DP103, a DEAD box-containing protein that assembles a novel corepressor complex. Functional interactions between the METS/DP103 complex and E2F/ pRB family proteins are also necessary for inhibition of cellular proliferation, suggesting a combinatorial code that directs permanent cell cycle exit during terminal differentiation.     

Characteristics of the haemoproteid community in an expanding white-winged dove population

The haemoproteid community of 171 eastern white-winged doves (Zenaida asiatica asiatica) from the expanding Texas population was examined using thin blood smears. During summer 1997, heart blood was taken from doves within their historical breeding range (Lower Rio Grande Valley of Texas), an intermediate region (San Antonio and surrounding area), and the new breeding periphery (north central to southeast Texas). Two species were found: Haemoproteus columbae and Haemoproteus sacharovi. Infracommunities rarely occurred in heart blood, as only 20 of 132 infected doves demonstrated gametocytes of both species. Overall prevalence of H. columbae and H. sacharovi was 77 and 15%, respectively. Prevalence of H. columbae was higher in the Lower Rio Grande Valley (LRGV) and intermediate regions than at the periphery, higher in adults than juveniles, and similar between males and females. Prevalence of H. sacharovi was lower in the LRGV than intermediate and peripheral regions, similar between juveniles and adults, and higher in females than males. Mean density of H. columbae and H. sacharovi was 15.9 +/- 2.7 and 0.3 +/- 0.1 (mean +/- SE per 3,000 erythrocytes), respectively. Overall mean abundance of H. columbae and H. sacharovi was 12.2 +/- 2.2 and 0.04 +/- 0.02, respectively. Mean abundance of H. columbae was higher in the LRGV and intermediate regions than at the periphery and was similar between host age and between host sex; H. sacharovi was similar among regions, host age, and host sex. This study emphasizes the importance of using prevalence, density, and abundance data to assess haemoproteid community structure and pattern.     

Determination of olanzapine in human blood by liquid chromatography-tandem mass spectrometry

A liquid chromatographic-tandem mass spectrometric (LC-MS-MS) assay was developed and validated to quantitatively determine olanzapine (OLZ) concentrations in human blood. Liquid-liquid extraction, using n-butanol:cyclohexane (3:47, v/v), was used to isolate OLZ and its internal standard, LY170158, from the biological matrix. Chromatographic resolution of OLZ from endogenous interferences and known metabolites was accomplished with a MetaChem Monochrom HPLC column (4.6 x 150 mm, d(p) 5 microm). Detection occurred using a Perkin-Elmer Sciex API III Plus triple quadrupole mass spectrometer using positive ion APCI and multiple reaction monitoring (MRM). The linear dynamic range was from 5 to 500 ng ml(-1) based on a 0.25-ml aliquot of human blood. The inter-day precision (%RSD) and accuracy (%RE) ranged from 3.65 to 10.64 and from -2.14 to 3.07, respectively. Modifications to an existing assay for the determination of OLZ in human plasma were necessary. A different structural analog was used as the internal standard due to instability observed for the original analog when using human blood as the matrix. A second modification was the addition of the anti-oxidant sodium ascorbate to inhibit degradation of OLZ in human blood, as has been noted by other investigators. Upon fortification of human blood with sodium ascorbate (final concentration, 0.33 mM), OLZ was found to be stable for at least 1 week at -70 degrees C as well as through two freeze-thaw cycles. This assay, which will be used to investigate the distribution of OLZ in human blood, grants insight into the proper sample handling conditions needed to perform valid determinations of OLZ in human blood.     

Signalling pathways that mediate skeletal muscle hypertrophy and atrophy

Atrophy of skeletal muscle is a serious consequence of numerous diseases, including cancer and AIDS. Successful treatments for skeletal muscle atrophy could either block protein degradation pathways activated during atrophy or stimulate protein synthesis pathways induced during skeletal muscle hypertrophy. This perspective will focus on the signalling pathways that control skeletal muscle atrophy and hypertrophy, including the recently identified ubiquitin ligases muscle RING finger 1 (MuRF1) and muscle atrophy F-box (MAFbx), as a basis to develop targets for pharmacologic intervention in muscle disease.     

Sequence and transfection of BoLA-DRB3 cDNAs

Bovine MHC (BoLA-) DRB3 alleles encoded by the DH8A, DH22A and DH24A class II haplotypes were cloned from cDNA and characterized by sequence analysis. Comparison with other full-length DRB3 sequences suggested that DRB3 alleles may have evolved through multiple lineages. All three BoLA-DRB3 alleles were shown to express on the surface of transfected cells, and the transfectants were used to define or confirm the class II specificity of a panel of monoclonal antibodies.