Artificial Induction and Evaluation of a Mild Isolate of Tomato Spotted Wilt Virus

A severe isolate (BL) of tomato spotted wilt virus (TSWV) that originated from Hawaii was treated with nitrous acid in an effort to obtain mild mutants. The standardized procedure used in mutation experiments was: extracting infected Gomphrena globosa L. leaf tissue in 0.01 M Na₂SO₃, 0.125 M sodium acetate and 0.4 M sodium nitrite at pH 5.5 and incubating the extract for 20 min at room temperature. The extract was inoculated to tobacco (Nicotiana tabacum L. cv. Havana 423) and local lesions were subsequently transferred to lettuce (Lactuca sativa L. cv. Minetto). One isolate (R27G) that incited mild symptoms in lettuce was obtained out of 868 local-lesion-transfers. Under greenhouse conditions, the isolate induced mild symptoms on tomato (Lycopersicon esculentum Mill.) but was severe on peppers (Capsicum annuum L.). The effect of the R27G isolate on growth of potted tomatoes kept outdoors was variable. In one trial, only 15 % of the fruit had symptoms versus 67 % in another trial. R27G fully protected Datura stramonium L. plants that were challenge inoculated with the severe parent BL isolate. Less effective cross protection was observed against a severe isolate from Oklahoma.     

Effect of mersalyl on mitochondrial Mg++ flux

The mercurial mersalyl has little effect either on rapid Mg⁺⁺ binding by isolated rat liver mitochondria or on the total Mg⁺⁺ content of these organelles measured after 0.75 min of incubation at 20°C. The data do not support the previous suggestion that the increased permeability to K⁺ of mitochondria treated with mersalyl results from release of endogenous Mg⁺⁺. An increased pH-dependence of unidirectional Mg⁺⁺ flux into respiring rat liver mitochondria is suggested to arise indirectly from inhibition by mersalyl of pH shifts associated with exchanges of endogenous phosphate. In addition, mersalyl appears to have a stimulatory effect on Mg⁺⁺ influx. Mersalyl also increases the average rate of unidirectional efflux of endogenous Mg⁺⁺. The stimulatory effects of mersalyl on Mg⁺⁺ flux are similar to, although quantitatively less than, the previously reported effects of mersalyl on mitochondrial K⁺ flux.     

Ultrasound‐mediated synthesis of camphoric acid‐based chiral salens for the enantioselective trimethylsilylcyanation of aldehydes

New chiral salen ligands were prepared by the ultrasound‐irradiated condensation of optically active (1R, 3S)‐1,2,2‐trimethyl‐1,3‐diaminocyclopentane with aromatic 1‐hydroxyaldehydes. The ultrasound‐mediated process is more convenient due to shorter reaction times, energy economy, and easier isolation of the products. The in situ formed Ti(IV)(salen) complexes, evaluated as catalysts in the enantioselective trimethylsilylcyanation of benzaldehyde, were found to be efficient for this process, originating the corresponding product in high yields (72–99%) and selectivities of up to 79%. The lowest energy transition states were determined by computational studies. These results were in qualitative agreement with the experimentally observed ones. Chirality, 2010. © 2009 Wiley‐Liss, Inc.     

The IGF1 small dog haplotype is derived from Middle Eastern grey wolves: a closer look at statistics,sampling, and the alleged Middle Eastern origin of small dogs

This paper is a response to Gray MM, Sutter NB, Ostrander EA, Wayne RK: The IGF1 small dog haplotype is derived from Middle Eastern grey wolves. BMC Biology 2010, 8:16.     

An overview of malaria transmission from the perspective of Amazon Anopheles vectors

In the Americas, areas with a high risk of malaria transmission are mainly located in the Amazon Forest, which extends across nine countries. One keystone step to understanding the Plasmodium life cycle in Anopheles species from the Amazon Region is to obtain experimentally infected mosquito vectors. Several attempts to colonise Ano- pheles species have been conducted, but with only short-lived success or no success at all. In this review, we review the literature on malaria transmission from the perspective of its Amazon vectors. Currently, it is possible to develop experimental Plasmodium vivax infection of the colonised and field-captured vectors in laboratories located close to Amazonian endemic areas. We are also reviewing studies related to the immune response to P. vivax infection of Anopheles aquasalis, a coastal mosquito species. Finally, we discuss the importance of the modulation of Plasmodium infection by the vector microbiota and also consider the anopheline genomes. The establishment of experimental mosquito infections with Plasmodium falciparum, Plasmodium yoelii and Plasmodium berghei parasites that could provide interesting models for studying malaria in the Amazonian scenario is important. Understanding the molecular mechanisms involved in the development of the parasites in New World vectors is crucial in order to better determine the interaction process and vectorial competence.     

Bacterial survival studies to assess the efficacy of static pile composting and above ground burial for disposal of bovine carcases

Aim: To assess the survival of bacteria during two alternative means of cattle carcase disposal in windrows: static pile composting (SPC) and above ground burial in soil (AGB), under temperate climate conditions on agricultural land, compared to surface disposal as the control method. Methods and Results: Bacteriological reference materials (pooled bovine faeces in permeable nylon bags and lyophilized cultures of Escherichia coli in glass ampoules) were positioned above and below each of 33 beef cattle carcases (250–300 kg). Temperatures at these sites were monitored with data loggers, while temperature and CO₂ probes were applied repeatedly at varying depths along the windrows. Aliquots of each reference material were cultured from three randomly selected animals from the SPC and AGB group and from all three control animals on five occasions (at 28, 56, 84, 126 and 182 days). SPC was highly efficacious in the destruction of coliforms in faeces and E. coli in ampoules within 28 days, while AGB was not significantly better than controls until 84 days, and bacteria in reference materials above the AGB carcases were still viable after 182 days. Temperature probes and loggers showed SPC provided sustained temperatures of 55–70°C, while AGB did not reach temperatures of 30°C, and the temperature differences correlated with bacteriological findings. Conclusions: In relation to emergency disease management, SPC can be successfully applied to eliminate pathogenic bacteria in cattle carcases, but AGB is unsuitable for carcase disposal. Significance and Impact of the Study: In emergency, animal disease outbreaks in temperate climates requiring large‐scale ruminant carcase disposal, SPC can be successfully applied for the destruction of micro‐organisms.     

An IGF-I promoter polymorphism modifies the relationships between birth weight and risk factors for cardiovascular disease and diabetes at age 36

BACKGROUND: Biochemical testing for pheochromocytoma by measurement of fractionated plasma metanephrines is limited by false positive rates of up to 18% in people without known genetic predisposition to the disease. The plasma normetanephrine fraction is responsible for most false positives and plasma normetanephrine increases with age. The objective of this study was to determine if we could improve the specificity of fractionated plasma measurements, by statistically adjusting for age. METHODS: An age-adjusted metanephrine score was derived using logistic regression from 343 subjects (including 33 people with pheochromocytoma) who underwent fractionated plasma metanephrine measurements as part of investigations for suspected pheochromocytoma at Mayo Clinic Rochester (derivation set). The performance of the age-adjusted score was validated in a dataset of 158 subjects (including patients 23 with pheochromocytoma) that underwent measurements of fractionated plasma metanephrines at Mayo Clinic the following year (validation dataset). None of the participants in the validation dataset had known genetic predisposition to pheochromocytoma. RESULTS: The sensitivity of the age-adjusted metanephrine score was the same as that of traditional interpretation of fractionated plasma metanephrine measurements, yielding a sensitivity of 100% (23/23, 95% confidence interval [CI] 85.7%, 100%). However, the false positive rate with traditional interpretation of fractionated plasma metanephrine measurements was 16.3% (22/135, 95% CI, 11.0%, 23.4%) and that of the age-adjusted score was significantly lower at 3.0% (4/135, 95% CI, 1.2%, 7.4%) (p < 0.001 using McNemar's test). CONCLUSION: An adjustment for age in the interpretation of results of fractionated plasma metanephrines may significantly decrease false positives when using this test to exclude sporadic pheochromocytoma. Such improvements in false positive rate may result in savings of expenditures related to confirmatory imaging.     

Elevated levels of fibrinogen-derived endogenous citrullinated peptides in synovial fluid of rheumatoid arthritis patients

Introduction

Rheumatoid arthritis (RA) is an autoimmune disease characterized by inflammation of the joints and the presence of autoantibodies directed against proteins containing the non-standard arginine-derived amino acid citrulline. The protein fibrinogen, which has an essential role in blood clotting, is one of the most prominent citrullinated autoantigens in RA, particularly because it can be found in the inflamed tissue of affected joints. Here, we set out to analyze the presence of citrullinated endogenous peptides in the synovial fluid of RA and arthritic control patients.

Methods

Endogenous peptides were isolated from the synovial fluid of RA patients and controls by filtration and solid phase extraction. The peptides were identified and quantified using high-resolution liquid chromatography-mass spectrometry.

Results

Our data reveal that the synovial fluid of RA patients contains soluble endogenous peptides, derived from fibrinogen, containing significant amounts of citrulline residues and, in some cases, also phosphorylated serine. Several citrullinated peptides are found to be more abundantly present in the synovial fluid of RA patients compared to patients suffering from other inflammatory diseases affecting the joints.

Conclusions

The increased presence of citrullinated peptides in RA patients points toward a possible specific role of these peptides in the immune response at the basis of the recognition of citrullinated peptides and proteins by RA patient autoantibodies.     

Assaying for Pollen Drift from Transgenic ‘Rainbow’ to Nontransgenic ‘Kapoho’ Papaya under Commercial and Experimental Field Conditions in Hawaii

In 1992, papaya ringspot virus (PRSV) was discovered in the Puna district of Hawaii island where 95% of the state of Hawaii’s papaya was being grown. By 1998 production in Puna had decreased 50% from 1992 levels. A PRSV-resistant transgenic papaya ‘Rainbow’ containing the coat protein gene of PRSV was released commercially in Hawaii in 1998, and saved the papaya industry from further devastation. In the ensuing years since the release of the transgenic papaya, a number of farmers grew hermaphrodite nontransgenic ‘Kapoho’ papaya in close proximity to plantings of hermaphrodite transgenic ‘Rainbow’ papaya. These plantings provided a unique opportunity to assay for transgenic-pollen drift under commercial conditions. Between 2004 and 2010, assays for the GUS (beta-glucuronidase) transgene in embryos were done to study transgenic-pollen drift in commercial ‘Kapoho’ plantings and in replicated field plots. Very low pollen drift (0.8%) was detected in fruit of ‘Kapoho’ trees in the border row of one plantation when 90 embryos were assayed per fruit, while no pollen drift was detected in four other commercial plantings in which eight embryos were tested per fruit. Pollen drift averaged 1.3% of tested embryos in field plots where individual hermaphrodite ‘Kapoho’ trees were adjacent to two or four ‘Rainbow’ trees. In contrast, 67.4% of tested embryos were GUS positive in similarly located female ‘Kapoho’ trees. The very low transgene flow to close-by ‘Kapoho’ plantings is likely due to the fact that hermaphrodite trees are used commercially in Hawaii and that these trees are largely self-pollinated before the stigma is exposed to external pollen.