Exertional Rhabdomyolysis in an Austere Deployed Setting.

INTRODUCTION: Exertional rhabdomyolysis is an, unfortunately, common disease process of military members undergoing strenuous activities. In an austere location, decisions on management and transfer to higher levels of care must be considered. This case series aims to discuss care in more austere locations. MATERIALS AND METHODS: A case series of 3 patients treated at Craig Joint Theater Hospital, Afghanistan. RESULTS: All patients were successfully treated with intravenous hydration and resumed normal military functions in theater rapidly. CONCLUSION: Management can be done at austere locations if certain labs and intravenous therapy are available and patients can be returned to normal duty quickly on the basis of symptom resolution.

Differentiation Among Blumeria graminis f. sp. tritici Isolates Originating from Wild Versus Domesticated Triticum Species in Israel.

Israel and its vicinity constitute a center of diversity of domesticated wheat species (Triticum aestivum and T. durum) and their sympatrically growing wild relatives, including wild emmer wheat (T. dicoccoides). We investigated differentiation within the forma specialis of their obligate powdery mildew pathogen, Blumeria graminis f. sp. tritici. A total of 61 B. graminis f. sp. tritici isolates were collected from the three host species in four geographic regions of Israel. Genetic relatedness of the isolates was characterized using both virulence patterns on 38 wheat lines (including 21 resistance gene differentials) and presumptively neutral molecular markers (simple-sequence repeats and single-nucleotide polymorphisms). All isolates were virulent on at least some genotypes of all three wheat species tested. All assays divided the B. graminis f. sp. tritici collection into two distinct groups, those from domesticated hosts and those from wild emmer wheat. One-way migration was detected from the domestic wheat B. graminis f. sp. tritici population to the wild emmer B. graminis f. sp. tritici population at a rate of five to six migrants per generation. This gene flow may help explain the overlap between the distinct domestic and wild B. graminis f. sp. tritici groups. Overall, B. graminis f. sp. tritici is significantly differentiated into wild-emmer and domesticated-wheat populations, although the results do not support the existence of a separate f. sp. dicocci.

Structure and Migration in U.S. Blumeria graminis f. sp. tritici Populations.

While wheat powdery mildew occurs throughout the south-central and eastern United States, epidemics are especially damaging in the Mid-Atlantic states. The structure of the U.S. Blumeria graminis f. sp. tritici population was assessed based on a sample of 238 single-spored isolates. The isolates were collected from 16 locations in 12 states (18 site-years) as chasmothecial samples in 2003 or 2005, or as conidial samples in 2007 or 2010. DNA was evaluated using nine single nucleotide polymorphism (SNP) markers in four housekeeping genes, and 10 simple sequence repeat (SSR) markers. The SSR markers were variably polymorphic, with allele numbers ranging from 3 to 39 per locus. Genotypic diversity was high (210 haplotypes) and in eight of the site-years, every isolate had a different SSR genotype. SNP haplotypic diversity was lower; although 15 haplotypes were identified, the majority of isolates possessed one of two haplotypes. The chasmothecial samples showed no evidence of linkage disequilibrium (P = 0.36), while the conidial samples did (P = 0.001), but the two groups had nearly identical mean levels of genetic diversity, which was moderate. There was a weakly positive relationship between genetic distance and geographic distance (R(2) = 0.25, P = 0.001), indicating modest isolation by distance. Most locations in the Mid-Atlantic and Great Lakes regions clustered together genetically, while Southeast locations formed a distinct but adjacent cluster; all of these were genetically separated from Southern Plains locations and an intermediate location in Kentucky. One-way migration was detected at a rate of approximately five individuals per generation from populations west of the Appalachian Mountains to those to the east, despite the fact that the Atlantic states experience more frequent and damaging wheat mildew epidemics. Overall, the evidence argues for a large-scale mosaic of overlapping populations that re-establish themselves from local sources, rather than continental-scale extinction and re-establishment, and a low rate of long-distance dispersal roughly from west to east, consistent with prevailing wind directions.

Molecular characterization of a new powdery mildew resistance gene Pm54 in soft red winter wheat.

KEY MESSAGE: A new powdery mildew resistance gene Pm54 was identified on chromosome 6BL in soft red winter wheat. Powdery mildew is causing increasing damage to wheat production in the southeastern USA. To combat the disease, a continuing need exists to discover new genes for powdery mildew resistance and to incorporate those genes into breeding programs. Pioneer(®) variety 26R61 (shortened as 26R61) and AGS 2000 have been used as checks in the Uniform Southern Soft Red Winter Wheat Nursery for a decade, and both have provided good resistance across regions during that time. In the present study, a genetic analysis of mildew resistance was conducted on a RIL population developed from a cross of 26R61 and AGS 2000. Phenotypic evaluation was conducted in the field at Plains, GA, and Raleigh, NC, in 2012 and 2013, a total of four environments. Three quantitative trait loci (QTL) with major effect were consistently detected on wheat chromosomes 2BL, 4A and 6BL. The 2BL QTL contributed by 26R61 was different from Pm6, a widely used gene in the southeastern USA. The other two QTL were identified from AGS 2000. The 6BL QTL was subsequently characterized as a simple Mendelian factor when the population was inoculated with a single Blumeria graminis f. sp. tritici (Bgt) isolate in controlled environments. Since there is no known powdery mildew resistance gene (Pm) on this particular location of common wheat, the gene was designated Pm54. The closely linked marker Xbarc134 was highly polymorphic in a set of mildew differentials, indicating that the marker should be useful for pyramiding Pm54 with other Pm genes by marker-assisted selection.

Rye-derived powdery mildew resistance gene Pm8 in wheat is suppressed by the Pm3 locus.

Genetic suppression of disease resistance is occasionally observed in hexaploid wheat or in its interspecific crosses. The phenotypic effects of genes moved to wheat from relatives with lower ploidy are often smaller than in the original sources, suggesting the presence of modifiers or partial inhibitors in wheat, especially dilution effects caused by possible variation at orthologous loci. However, there is little current understanding of the underlying genetics of suppression. The discovery of suppression in some wheat genotypes of the cereal rye chromosome 1RS-derived gene Pm8 for powdery mildew resistance offered an opportunity for analysis. A single gene for suppression was identified at or near the closely linked storage protein genes Gli-A1 and Glu-A3, which are also closely associated with the Pm3 locus on chromosome 1AS. The Pm3 locus is a complex of expressed alleles and pseudogenes embedded among Glu-A3 repeats. In the current report, we explain why earlier work indicated that the mildew suppressor was closely associated with specific Gli-A1 and Glu-A3 alleles, and predict that suppression of Pm8 involves translated gene products from the Pm3 locus.

Population genetic analysis of an Eastern U.S. wheat powdery mildew population reveals geographic subdivision and recent common ancestry with U.K. and Israeli populations.

The structure of the U.S. wheat powdery mildew population (Blumeria graminis f. sp. tritici) has not been previously investigated, and the global evolutionary history of B. graminis f. sp. tritici is largely unknown. After gathering 141 single-ascosporic B. graminis f. sp. tritici isolates from 10 eastern U.S. locations, 34 isolates from the United Kingdom, and 28 isolates from Israel, we analyzed pathogen population structure using presumptively neutral markers. DNA was extracted from conidia, primers for 12 "housekeeping" genes were designed, and amplicons were examined for polymorphism. Four genes were found to contain a total of 12 single-nucleotide polymorphisms in the U.S. population and were also analyzed in the U.K. and Israeli populations. In total, 25 haplotypes were inferred from the four concatenated genes, with 2 haplotypes comprising over 70% of the U.S. population. Using Hudson's tests and analysis of molecular variance, we found the wheat mildew isolates subdivided into four groups corresponding to distinct regions: the mid-Atlantic United States, the southern United States, the United Kingdom, and Israel. Genotypic diversity was greatest in samples from the United Kingdom, Israel, Virginia, and Kingston, NC. Using rarefaction, a procedure that compensates for differing sample sizes when estimating population richness and diversity, we found that cooler locations with greater conduciveness to regular powdery mildew epidemics had the greatest haplotype richness. Our results suggest that the eastern U.S. B. graminis f. sp. tritici population is young, descended recently from Old World populations with isolation and genetic drift, and is currently subdivided into northern and southern subpopulations.

Virulence Structure of the Eastern U.S. Wheat Powdery Mildew Population.

Little is known about the population structure of wheat powdery mildew in the eastern United States, and the most recent report on virulence in this population involved isolates collected in 1993-94. In the present study, wheat leaves naturally infected with powdery mildew were collected from 10 locations in the southeastern United States in 2003 and 2005 and a collection of 207 isolates was derived from single ascospores. Frequencies of virulence to 16 mildew resistance (Pm) genes were determined by inoculating the isolates individually on replicated plates of detached leaves of differential wheat lines. These virulence frequencies were used to infer local effectiveness of Pm genes, estimate virulence complexity, detect significant associations between pairs of pathogen avirulence loci, and assess whether phenotypic differences between pathogen subpopulations increased with geographic distance. In both years, virulence to Pm3a, Pm3c, Pm5a, and Pm7 was present in more than 90% of sampled isolates and virulence to Pm1a, Pm16, Pm17, and Pm25 was present in fewer than 10% of isolates. In each year, 71 to 88% of all sampled isolates possessed one of a few multilocus virulence phenotypes, although there were significant differences among locations in frequencies of virulence to individual Pm genes. Several significant associations were detected between alleles for avirulence to pairs of Pm genes. Genetic (phenotypic) distance between isolate subpopulations increased significantly (R2 = 0.40, P < 0.001) with increasing geographic separation; possible explanations include different commercial deployment of Pm genes and restricted gene flow in the pathogen population.

The redox behavior of the heme in cystathionine beta-synthase is sensitive to pH.

Human cystathionine beta-synthase (CBS) is a unique pyridoxal-5'-phosphate-dependent enzyme in which heme is also present as a cofactor. Because the function of heme in this enzyme has yet to be elucidated, the study presented herein investigated possible relationships between the chemistry of the heme and the strong pH dependence of CBS activity. This study revealed, via study of a truncation variant, that the catalytic core of the enzyme governs the pH dependence of the activity. The heme moiety was found to play no discernible role in regulating CBS enzyme activity by sensing changes in pH, because the coordination sphere of the heme is not altered by changes in pH over a range of pH 6-9. Instead, pH was found to control the equilibrium amount of ferric and ferrous heme present after reaction of CBS with one-electron reducing agents. A variety of spectroscopic techniques, including resonance Raman, magnetic circular dichroism, and electron paramagnetic resonance, demonstrated that at pH 9 Fe(II) CBS is dominant while at pH 6 Fe(III) CBS is favored. At low pH, Fe(II) CBS forms transiently but reoxidizes by an apparent proton-gated electron-transfer mechanism. Regulation of CBS activity by the iron redox state has been proposed as the role of the heme moiety in this enzyme. Given that the redox behavior of the CBS heme appears to be controlled by pH, interplay of pH and oxidation state effects must occur if CBS activity is redox regulated.

Investigation of the role of the N-terminal proline, the distal heme ligand in the CO sensor CooA.

A unique feature of CooA, a heme-containing transcription factor, is that the N-terminal proline is the distal heme ligand in the ferrous state, and this ligand is displaced upon CO binding. To investigate the importance of Pro(2) in CO-dependent DNA binding, several CooA variants that alter N-terminal ligation were characterized. Electronic absorption, electron paramagnetic resonance, and magnetic circular dichroism spectra of these variants provide the most definitive evidence that Pro(2) is the distal ligand in Fe(III) CooA. Furthermore, the functional and spectroscopic properties of these proteins depended on whether a weak ligand occupied the distal heme coordination site: for CooA variants in which distal coordination is disrupted, the DNA-binding affinities and Fe(II)-CO spectral properties showed an unexpected dependence on the order of CO addition and heme reduction. If N-terminal variant samples were incubated with CO before the heme was reduced, the proteins displayed DNA-binding affinities and Fe(II)-CO spectral characteristics similar to those of wild-type (WT) CooA. However, if the same samples were incubated with CO after the heme was reduced, the extent of functional and spectral similarity to WT CooA negatively correlated with the amount of high-spin heme present in the ferric state. From these data, it was inferred that the absence of a distal heme ligand in the ferric state prevents WT-like CO binding to the ferrous state, and it was hypothesized that correct CO binding is inhibited by the collapse of the distal heme pocket upon reduction. Together with the observation that L116H CooA, a variant in which His(116) replaces Pro(2) as the distal heme ligand, binds CO more slowly than WT CooA, these data indicate that the presence of a weak distal heme ligand, not specifically ligation by the N-terminal proline, is crucial for proper function. The role of Pro(2) in CooA is apparently to direct CO to bind on the distal side of heme and to help maintain the integrity of the distal heme pocket during the redox-mediated ligand switch.