Identification and functional characterization of genes involved in the sexual reproduction of the ascomycete fungus Gibberella zeae.
We previously reported that G protein alpha subunit 1 (GPA1) is essential for sexual reproduction in the homothallic ascomycete fungus Gibberella zeae. In this study we performed microarray analyses on a GPA1 deletion mutant of G. zeae (Δgpa1) to identify genes involved in the sexual reproduction of this fungus. In the Δgpa1 strain, 645 genes were down-regulated and 550 genes were up-regulated during sexual reproduction when compared to the wild-type strain. One hundred of the down-regulated genes were selected for further investigation based on orthologous group clusters and differences in transcript levels. Quantitative real time-PCR was used to determine transcriptional profiles of these genes at various sexual and vegetative stages. We observed that transcript levels of 78 of these genes were dramatically increased in the wild-type strain during sexual reproduction compared to levels observed during vegetative growth, and were down-regulated in Δgpa1 compared to the wild-type strain. We deleted 57 of these genes and found that four of the deletion mutants lost self-fertility and five produced fewer perithecia compared to the wild-type strain. Two mutants produced wild-type numbers of perithecia, but maturation of perithecia and ascospores was delayed. In all we identified 11 genes that are involved in sexual reproduction of G. zeae and present evidence that some of these genes function at distinct stages during sexual reproduction in the fungus.

title: Population structure of and mycotoxin production by Fusarium graminearum from maize in South Korea.
abstract: Fusarium graminearum (Gibberella zeae) is an important pathogen of wheat, maize, barley, and rice in South Korea, and harvested grain often is contaminated with trichothecenes such as deoxynivalenol and nivalenol. In this study, we examined 568 isolates of F. graminearum collected from maize at eight locations in South Korea. We used amplified fragment length polymorphisms (AFLPs) to identify four lineages (2, 3, 6, and 7); lineage 7 was the most common (75%), followed by lineage 6 (12%), lineage 3 (12%), and lineage 2 (1%). The genetic identity among populations was high (>0.98), and the effective migration rate between locations was higher than that between lineages. Female fertility varied by lineage: all lineage 7 isolates were fertile, while 70%, 26%, and 14% of the isolates in lineages 6, 3, and 2, respectively, were fertile. All lineage 3 and lineage 7 isolates produced deoxynivalenol, whereas most lineage 2 and 6 isolates produced nivalenol. Genotypic diversity in lineage 3 and lineage 6 populations is similar to that found in previously described Korean rice populations, but genotypic diversity in lineage 7 is much lower, even though similar levels of gene flow occur between lineage 7 populations. We conclude that lineage 7 was relatively recently introduced into South Korea, perhaps accompanying imported maize seeds.

title: Rapid determination of bacteria in pools.
abstract: If a method for rapid determination of bacterial levels in swimming pools and whirlpools were available, bacterial health risks could be avoided before exposure occurred. Such a method would enable public-health authorities to make timely decisions about closing or reopening pools and to adjust disinfectant doses. The purpose of this study was to determine if a rapid adenosine triphosphate (ATP) bioluminescence method would serve this purpose. Bacteriological analyses were completed on samples from swimming pools and whirlpools in Washtenaw County, Michigan. The results from the ATP assay were compared with those from the conventional plate count method and were found to be highly correlated (r = .92). To assess whether a prefiltration step was necessary to delete nonbacterial ATP--which might originate from bathers' skin cells or other debris--two procedures were tested: one with prefiltration and one without prefiltration. The results for prefiltered samples were not statistically different from the results for non-prefiltered samples (p > .05), for both the heterotrophic plate counts and the ATP bioluminescence method. This study indicates that the ATP assay can estimate bacterial levels in pools within minutes and without prefiltration.

title: Activation of Ets-2 by oxidative stress induces Bcl-xL expression and accounts for glial survival in amyotrophic lateral sclerosis.
abstract: Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disease characterized by selective degeneration of motor neurons and glial activation. Cell-specific transcriptional regulation induced by oxidative stress may contribute to the survival and activation of astrocytes in the face of motor neuron death. In the present study, we demonstrate an age-dependent increase in Bcl-xL and Ets-2 immunoreactivity that correlates with an increase of glial fibrillary acidic protein (GFAP)-positive cells in the ventral horn of the spinal cord in both ALS transgenic mice [mutant SOD1 (G93A)] and affected humans. Chromatin immunoprecipitation (ChIP) analysis verified that Ets-2 preferentially occupies the Ets-2 binding element in the promoter of Bcl-xL in primary astrocytes under oxidative stress conditions as well as in G93A spinal cords. Ets-2 small-interfering RNA down-regulated the transcriptional activity of Bcl-xL. In primary glial cultures, Bcl-xL overexpression and mutant SOD1 (G93A) both conferred resistance to oxidative stress-induced cell death. Our findings suggest that Ets-2 transcription factor activation of Bcl-xL gene may protect glia from constitutive oxidative stress that is thought to be a key mechanism contributing to the pathogenesis of ALS. This survival pathway may contribute to the glial survival and activation seen in the spinal cord of ALS patients.

title: Anthocyanins and other polyphenolics in American elderberry (Sambucus canadensis) and European elderberry (S. nigra) cultivars.
abstract: Ten genotypes representing two elderberry species, Sambucus canadensis L. (eight genotypes) and S. nigra L. (two genotypes), were examined for their anthocyanins (ACY), total phenolics (TP),°Brix, titratable acidity (TA), and pH over two growing seasons.



Overall, fruit generally had higher ACY, TP, ACY/TP,°Brix, and pH in 2005 than 2004. All samples of S. canadensis had similar anthocyanin profiles to one another, but were distinctly different from S. nigra. Both species had cyanidin-based anthocyanins as major pigments. Previously unreported anthocyanins were identified in some samples in this study. Trace levels of delphinidin 3-rutinoside were present in all elderberry samples except cv. 'Korsør'. Also, petunidin 3-rutinoside was detected in cvs 'Adams 2', 'Johns', 'Scotia', 'York', and 'Netzer' (S. canadensis). The identified polyphenolics of both species were mainly composed of cinnamic acids and flavonol glycosides. The major polyphenolic compounds present in S. canadensis were neochlorogenic acid, chlorogenic acid, rutin, and isorhamnetin 3-rutinoside, while chlorogenic acid and rutin were found to be major polyphenolic compounds in S. nigra.



Sufficient variability was seen among these genotypes to suggest that a successful breeding program could be carried out to improve levels of the various compounds evaluated in this study.

title: Missed opportunities in the treatment of pediatric appendicitis
abstract: IntroductionThis study sought to evaluate the outcomes of in-hospital delay and determine associated co-morbidities in the treatment of pediatric acute appendicitis.MethodsThis was a retrospective analysis of the national inpatient sample from 2000 to 2008. Immediate treatment was defined as treatment in hospital day 0 or 1. Delay in treatment was defined as treatment in hospital day 2 and beyond.ResultsDuring the study period, 683,016 pediatric appendicitis were identified. 17,737 (2.6 %) experienced a delay in treatment. Multivariate analysis identified associated co-morbidities for delay in treatment: ALL (OR 12.84, CI 11.04–14.94), AML (OR 9.41, CI 7.58–11.68), neutropenia (OR 5.53, CI 4.60–6.65), and ovarian cyst without torsion (OR 3.17, CI 2.94–3.42). Surgical management included more than 13-fold increase in drainage procedures (5.5 vs. 0.4 %), sixfold increase in cecectomy (1.2 vs. 0.2 %), 14-fold increase in hemicolectomy (1.4 vs. 0.1 %), 11-fold increase in small bowel laceration suture repair (1.1 vs. 0.1 %), and 15-fold increase in small bowel resection (1.5 vs. 0.1 %).ConclusionsIn-hospital delay beyond 2 days is associated with significant negative outcomes with regard to complications, economic burden, and subsequent surgical management. Using the co-morbidity index, high-risk co-morbidities with associated delay in treatment were identified.

title: Traction forces generated by locomoting keratocytes
abstract: Traction forces produced by moving fibroblasts have been observed as distortions in flexible substrata including wrinkling of thin, silicone rubber films. Traction forces generated by fibroblast lamellae were thought to represent the forces required to move the cell forwards. However, traction forces could not be detected with faster moving cell types such as leukocytes and growth cones (Harris, A. K., D. Stopak, and P. Wild. 1981.  ). We have developed a new assay in which traction forces produced by rapidly locomoting fish keratocytes can be detected by the two-dimensional displacements of small beads embedded in the plane of an elastic substratum. Traction forces were not detected at the rapidly extending front edge of the cell. Instead the largest traction forces were exerted perpendicular to the left and right cell margins. The maximum traction forces exerted by keratocytes were estimated to be approximately 2 x 10(-8) N. The pattern of traction forces can be related to the locomotion of a single keratocyte in terms of lamellar contractility and area of close cell- substratum contact.