First direct double-ß decay Q-value measurement of 82Se in support of understanding the nature of the neutrino.

In anticipation of results from current and future double-ß decay studies, we report a measurement resulting in a (82)Se double-ß decay Q value of 2997.9(3) keV, an order of magnitude more precise than the currently accepted value. We also present preliminary results of a calculation of the (82)Se neutrinoless double-ß decay nuclear matrix element that corrects in part for the small size of the shell model single-particle space. The results of this work are important for designing next generation double-ß decay experiments and for the theoretical interpretations of their observations.

The Armed Services Blood Program: blood support to combat casualty care 2001 to 2011.

BACKGROUND: The Armed Services Blood Program (ASBP) provides the farthest-reaching blood supply in the world. This article provides statistics and a review of blood operations in support of combat casualty care during the last 10 years. It also outlines changes in blood doctrine in support of combat casualty care. METHODS: This is a descriptive overview and review of blood product use and transfusions used by ASBP personnel to support combat operations in Iraq and Afghanistan between October 2001 and November 2011. RESULTS: The ASBP initiated major changes in blood availability and age of blood in theater. In support of data published by physicians in theater, showing improved patient survival when a higher ratio of fresh frozen plasma and red blood cells (RBCs) is achieved, plus the use of platelets, the ASBP increased availability of plasma and established platelet collection facilities in theater. New capabilities included emergency collection of apheresis platelets in the battlefield, availability and transfusion of deglycerolized red cells, rapid diagnostic donor screening, and a new modular blood detachment. Forward surgical facilities that were at one time limited to a blood inventory consisting of RBCs now have a complete arsenal of products at their fingertips that may include fresher RBCs, fresh frozen plasma, cryoprecipitate, and platelets. A number of clinical practice guidelines are in place to address these processes. Changes in blood doctrine were made to support new combat casualty care and damage-control resuscitation initiatives. CONCLUSION: Despite the challenges of war in two theaters of operation, a number of improvements and changes to blood policy have been developed during the last 10 years to support combat casualty care. The nature of medical care in combat operations will continue to be dynamic and constantly evolving. The ASBP needs to be prepared to meet future challenges. LEVEL OF EVIDENCE: Epidemiologic study, level IV.

Systemin and jasmonic acid regulate constitutive and herbivore-induced systemic volatile emissions in tomato, Solanum lycopersicum.

Transgenic tomato (Solanum lycopersicum) plants that overexpress the Prosystemin gene (35S::PS) and plants with a mutation in the JA biosynthetic pathway (def1) are known to exhibit a constitutive or reduced wound response, respectively. Here it is demonstrated that several independent 35S::PS lines emit high levels of specific volatiles in addition to increased accumulation of proteinase inhibitors (PIs). Furthermore, the temporal dynamics of systemically induced volatile compounds including green-leaf volatiles, terpenes, and shikimic acid-derivatives from 35S::PS and def1 plants in response to herbivore wounding and treatment with jasmonic acid (JA) are described. Application of JA induced defense protein accumulation and volatile emissions in wild type plants, but did not further increase systemic volatile emissions from 35S::PS plants. Wounding by Manduca sexta larvae induced synthesis of defense proteins and emission of volatiles in wild type plants, but not in def1 plants. Application of jasmonic acid restored the local and systemic accumulation of defense proteins in def1, as well as enhanced herbivore-induced volatile emissions. These results provide strong support for the role of prosystemin- and JA-signaling in the regulation of volatile emissions in tomato plants.

Thioredoxin and thioredoxin reductase expression in thyroid cancer depends on tumour aggressiveness.

Thyroid cancer is the second most common malignancy following breast cancer in Arab females. Thioredoxin (TRX) is a small multi-functional redox protein with both intracellular and extracellular functions. The protein exists in either a reduced form (thioredoxin-SH2) or an oxidized form (thioredoxin-S2). TRX acts as an enhancement for growth factors and stimulates the growth of cancer cells. In this study of thyroid neoplasms, involving 121 female and 62 male patients, expression of TRX and TRX-R was studied using purified mouse anti-human TRX monoclonal antibody and anti-human TRX-R antiserum from rabbits, respectively. In order to delineate tumour cell growth, proliferating cell nuclear antigen (PCNA) polyclonal antibody was used. Compared to normal thyroid tissue, expression of TRX and TRX-R was increased in the cytoplasm and nuclei of thyroid cancer cells. Furthermore, TRX expression correlated with that of TRX-R. Of the 183 thyroid neoplasms investigated, overexpression of TRX-R was found in different types of neoplasms. The majority of carcinomas showed a correlation between strongly positive TRX and TRX-R expression and neoplastic cellular proliferation, as measured by PCNA. This indicates that increased TRX and TRX-R expression may be associated with tumourigenesis by acting as an autocrine growth stimulus. This study suggests that TRX immunoreactivity in thyroid tumours is a function of malignancy and cancer progression. In addition, secreted TRX can also act as an extracellular growth factor for both normal and tumour cells and enhance the sensitivity of the cells. Furthermore, this study emphasizes the potential benefits of anti-TRX/TRX-R agents in cancer therapeutics in the treatment of thyroid cancer.

Production of the phytohormone indole-3-acetic acid by estuarine species of the genus Vibrio.

Strains of Vibrio spp. isolated from roots of the estuarine grasses Spartina alterniflora and Juncus roemerianus produce the phytohormone indole-3-acetic acid (IAA). The colorimetric Salkowski assay was used for initial screening of IAA production. Gas chromatography-mass spectroscopy (GC-MS) was then employed to confirm and quantify IAA production. The accuracy of IAA quantification by the Salkowski assay was examined by comparison to GC-MS assay values. Indole-3-acetamide, an intermediate in IAA biosynthesis by the indole-3-acetamide pathway, was also identified by GC-MS. Multilocus sequence typing of concatenated 16S rRNA, recA, and rpoA genes was used for phylogenetic analysis of environmental isolates within the genus Vibrio. Eight Vibrio type strains and five additional species-level clades containing a total of 16 environmental isolates and representing five presumptive new species were identified as IAA-producing Vibrio species. Six additional environmental isolates similar to four of the Vibrio type strains were also IAA producers. To our knowledge, this is the first report of IAA production by species of the genus Vibrio or by bacteria isolated from an estuarine environment.

Tomato MAPKs LeMPK1, LeMPK2, and LeMPK3 function in the systemin-mediated defense response against herbivorous insects.

Systemin is a wound-signaling peptide that mediates defenses of tomato plants against herbivorous insects. Perception of systemin by the membrane-bound receptor SR160 results in activation of MAPKs, synthesis of jasmonic acid (JA), and expression of defense genes. To test the function of MAPKs in the response to systemin, we used virus-induced gene silencing (VIGS) in plants that overexpress the systemin precursor prosystemin (35S::prosys plants). These transgenic plants accumulate high levels of defense proteins and exhibit increased resistance to herbivorous insects. Cosilencing of the MAPKs MPK1 and MPK2 reduced MPK1/2 kinase activity, JA biosynthesis, and expression of JA-dependent defense genes. Application of methyl-JA restored the full defense response. These data show that MPK1 and MPK2 are essential components of the systemin signaling pathway and most likely function upstream of JA biosynthesis. MPK1 and MPK2 are 95% identical at the amino acid level. Specific VIGS of only MPK1 or MPK2 resulted in the same reduction of defense gene expression as cosilencing of MPK1 and MPK2, indicating that gene dosage effects may be important for MPK signaling. In addition, VIGS of the closely related MPK3 also reduced systemin-induced defense responses. The function of MPK1/2 and orthologs in pathogen-induced defenses is well established. Here we show that cosilencing of MPK1 and MPK2 compromised prosystemin-mediated resistance to Manduca sexta (Lepidoptera) herbivory, demonstrating that MPK1 and MPK2 are also required for successful defenses against herbivorous insects.

Carbon inhibits vascular endothelial growth factor- and fibroblast growth factor-promoted angiogenesis.

Angiogenesis is important for normal growth and wound healing processes. An imbalance of the growth factors involved in this process, however, causes the acceleration of several diseases including malignant, ocular, and inflammatory diseases. Inhibiting angiogenesis through interfering with its pathway is a promising methodology to hinder the progression of these diseases. Herein, we studied the anti-angiogenic effects of various carbon materials such as graphite, multiwalled carbon nanotubes and fullerenes in vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF2)-induced angiogenesis evaluated in the chick chorioallantoic membrane (CAM) model. All the carbon materials tested showed substantial anti-angiogenic activity against either FGF2- or VEGF-induced angiogenesis in the CAM model. Those carbon materials did not have any significant effects on basal angiogenesis in the absence of the added growth factors.

Volatile emissions from an odorous plant in response to herbivory and methyl jasmonate exposure.

Induced volatile terpenes have been commonly reported among diverse agricultural plant species, but less commonly investigated in odorous plant species. Odorous plants synthesize and constitutively store relatively large amounts of volatiles, and these may play a role in defense against herbivores. We examined the effect of herbivory and methyl jasmonate (MeJA) exposure on the release of volatile organic compounds (VOCs) in the marsh elder, Iva frutescens, which contains numerous constitutive VOCs, mainly mono- and sesquiterpenes. Our specific goal was to test for the presence of inducible VOCs in a naturally occurring plant already armed with VOCs. The abundant, native specialist leaf beetle Paria aterrima was used in herbivore induction trials. VOCs were sampled from herbivore wounded and unwounded, and from MeJA treated and untreated I. frutescens. Total VOC emissions were significantly greater in response to herbivory and MeJA treatment compared to unwounded controls. Herbivore wounding caused a substantial shift in the emission profile (42 VOCs from wounded, compared to 8 VOCs from unwounded I. frutescens), and MeJA had a similar yet less substantial influence on the emission pattern (28 VOCs from MeJA treated compared to 8 VOCs from untreated I. frutescens). Constitutive VOC emissions predominated, but some VOCs were detected only in response to herbivory and MeJA treatment, suggesting de novo synthesis. Several VOCs exhibited a delayed emission profile in contrast to the rapid release of constitutive VOCs, and principal components analysis revealed they were not associated with constitutive emissions. While I. frutescens contains many constitutive VOCs that are released immediately in response to herbivory, it also produces novel VOCs in response to feeding by the specialist P. aterrima and MeJA treatment.

Spectroscopic characterization of the ferric states of Amphitrite ornata dehaloperoxidase and Notomastus lobatus chloroperoxidase: His-ligated peroxidases with globin-like proximal and distal properties.

Amphitrite ornata dehaloperoxidase (DHP) and Notomastus lobatus chloroperoxidase (NCPO) catalyze the peroxide-dependent dehalogenation of halophenols and halogenation of phenols, respectively. Both enzymes have histidine (His) as their proximal heme iron ligand. Crystallographic examination of DHP revealed that it has a globin fold [M.W. LaCount, E. Zhang, Y.-P. Chen, K. Han, M.M. Whitton, D.E. Lincoln, S.A. Woodin, L. Lebioda, J. Biol. Chem. 275 (2000) 18712-18716] and kinetics studies established that ferric DHP is the active state [R.L. Osborne, L.O. Taylor, K. Han, B. Ely, J.H. Dawson, Biochem. Biophys. Res. Commun. 324 (2004) 1194-1198]. NCPO likely has these same properties. Previous work with His-ligated heme proteins has revealed characteristic spectral distinctions between dioxygen binding globins and peroxide-activating peroxidases. Since DHP, and likely NCPO, is a peroxide-activating globin, we have sought to determine in the present investigation whether the ferric resting states of these two novel heme-containing enzymes are myoglobin-like or peroxidase-like. To do so, we have examined their exogenous ligand-free ferric states as well as their azide, imidazole and NO bound ferric adducts (and ferrous-NO complexes) with UV-Visible absorption and magnetic circular dichroism spectroscopy. We have also compared each derivative to the analogous states of horse heart myoglobin (Mb) and horseradish peroxidase (HRP). The spectra observed for parallel forms of DHP and NCPO are virtually identical to each other as well as to the spectra of the same Mb states, while being less similar to the spectra of corresponding HRP derivatives. From these data, we conclude that exogenous ligand-free ferric DHP and NCPO are six-coordinate with water and neutral His as ligands. This coordination structure is distinctly different from the ferric resting state of His-ligated peroxidases and indicates that DHP and NCPO do not activate bound peroxide through a mechanism dependent on a push effect imparted by a partially ionized proximal His as proposed for typical heme peroxidases.

The transcription factor Ets-1 in breast cancer.

The proto-oncogene Ets-1 is a member of the Ets family of transcription factors which share a unique DNA binding domain, the Ets domain. Ets binding sites have been described on the promoters or enhancers of many proteinases and several Ets members transcriptionally regulate such promoters in transient cotransfection assays. Ets-1 is involved in both normal and pathological functions. Ets-1 is expressed in a variety of cells, including endothelial cells, vascular smooth muscle cells and epithelial cells. Ets-1 regulates the expression of several angiogenic and extracellular matrix remodeling factors promoting an invasive phenotype. The Ets family of transcription factors may play a role in the disease progression of breast cancer. In tumors, including breast neoplasia, Ets-1 expression is indicative of poorer prognosis. This review will summarize the role of Ets-1 in both the tumor cells, and the tumor endothelial cells as it relates to breast tumor growth and spread.

Isolation of murine aortic endothelial cells in culture and the effects of sex steroids on their growth.

The lack of commercially available primary murine endothelial cells prompted us to isolate and cultivate this cell type. We report here the effect of sex steroids on the in vitro growth of murine aortic endothelial cells. Murine aortic endothelial cells were isolated by a combination of explant outgrowth from aortic rings and enzymatic digestion. The endothelial nature of the cells was verified by uptake of acylated low-density lipoprotein and positive staining for CD-31. Murine aortic endothelial cell growth is stimulated by physiological concentrations of estrogen. Progesterone, when given simultaneously with estrogen, inhibited the stimulatory growth effect of estrogen. Murine aortic endothelial cells grown in vitro continue to express messenger ribonucleic acid for proteins related to endothelial growth. These include vascular endothelial growth factor, its receptors Flt-1 and Flk-1, and the angiogenesis-associated transcription factor, Ets-1.

The thioredoxin-thioredoxin reductase system: over-expression in human cancer.

Redox control has emerged as a fundamental biological control mechanism. One of the major redox control systems is the thioredoxin system comprised of thioredoxin (TRX) and thioredoxin reductase (TR). Together they form a powerful system involved in many central intracellular and extracellular processes including cell proliferation, the redox regulation of gene expression and signal transduction, protection against oxidative stress, anti-apoptotic functions, growth factor and co-cytokine effects, and regulation of the redox state of the extracellular environment. Over recent years this system has increasingly been linked to the development and expression of cancer phenotypes. In this report immunocytochemical approaches have been used to simultaneously determine the expression and localisation of both TRX and TR in primary human cancers, including breast cancer, thyroid, prostate and colorectal carcinoma, and malignant melanoma. In aggressive invasive mammary carcinomas and advanced malignant melanomas, thioredoxin was highly over-expressed compared to tumours of lesser aggressive nature. TRX expression was found in both nuclear and cytoplasmic location in the neoplastic cells. Furthermore, increased levels of TRX positively correlate with thioredoxin reductase (TR) expression and localisation. These results, which are the first immunocytochemical studies on the in vivo expression and localisation of TRX and TR in melanomas, thyroid, prostate and colorectal carcinomas and the first reports of TR expression in breast carcinomas, significantly extend the range of human cancers for which such data is available. Overall the results support the conclusion that aggressive tumours greatly over-express both TRX and TR. Such tumours have a high proliferation capacity, a low apoptosis rate and an elevated metastatic potential strongly implicating the involvement of the TRX system in the processes of oncogenesis and tumourogenesis and confirming its potential as a target for anticancer therapy for a wide range of human tumours.

Development of gypsy moth larvae feeding on red maple saplings at elevated CO2 and temperature.

Predicted increases in atmospheric CO(2) and global mean temperature may alter important plant-insect associations due to the direct effects of temperature on insect development and the indirect effects of elevated temperature and CO(2) enrichment on phytochemicals important for insect success. We investigated the effects of CO(2) and temperature on the interaction between gypsy moth (Lymantria dispar L.) larvae and red maple (Acer rubrum L.) saplings by bagging first instar larvae within open-top chambers at four CO(2)/temperature treatments: (1) ambient temperature, ambient CO(2), (2) ambient temperature, elevated CO(2) (+300 microl l(-1) CO(2)), (3) elevated temperature (+3.5 degrees C), ambient CO(2), and (4) elevated temperature, elevated CO(2). Larvae were reared to pupation and leaf samples taken biweekly to determine levels of total N, water, non-structural carbohydrates, and an estimate of defensive phenolic compounds in three age classes of foliage: (1) immature, (2) mid-mature and (3) mature. Elevated growth temperature marginally reduced (P <0.1) leaf N and significantly reduced ( P <0.05) leaf water across CO(2) treatments in mature leaves, whereas leaves grown at elevated CO(2) concentration had a significant decrease in leaf N and a significant increase in the ratio of starch:N and total non-structural carbohydrates:N. Leaf N and water decreased and starch:N and total non-structural carbohydrates:N ratios increased as leaves aged. Phenolics were unaffected by CO(2) or temperature treatment. There were no interactive effects of CO(2) and temperature on any phytochemical measure. Gypsy moth larvae reached pupation earlier at the elevated temperature (female =8 days, P <0.07; male =7.5 days, P <0.03), whereas mortality and pupal fresh weight of insects were unrelated to either CO(2), temperature or their interaction. Our data show that CO(2) or temperature-induced alterations in leaf constituents had no effect on insect performance; instead, the long-term exposure to a 3.5 degrees C increase in temperature shortened insect development but had no effect on pupal weight. It appears that in some tree-herbivorous insect systems the direct effects of an increased global mean temperature may have greater consequences for altering plant-insect interactions than the indirect effects of an increased temperature or CO(2) concentration on leaf constituents.

Estrogen-induced Ets-1 promotes capillary formation in an in vitro tumor angiogenesis model.

We employed an in vitro angiogenesis model that simulates the in vivo milieu for tumor capillary formation to study the direct effects of estrogen. 17beta-estradiol (E2) treatment significantly stimulated capillary sprouting within 8 h in co-cultures of rat aortic endothelial cells (RAECs) and mouse mammary tumor cells. Co-cultures treated with either progesterone (P4) or E2+P4 showed minimal endothelial cell (EC) sprouting when compared to E2 treated cultures. Treatment with the E2 agonist ICI 182,780 dramatically inhibited capillary formation, demonstrating E2-specificity. Within hours, of E2 treatment ECs isolated from tumor cell/EC co-cultures demonstrated a statistically significant increase in both mRNA and protein levels of the transcription factor Ets-1. We observed increased matrix metalloproteinase (MMP) and decreased tissue inhibitor of metalloproteinase (TIMP) mRNA levels in these ECs following E2 treatment. Ets-1 upregulates expression of the vascular endothelial growth factor (VEGF) receptor, Flt-1 and we detected increased Flt-1 mRNA levels in ECs co-cultured with tumor cells following E2 treatment. Expression of Ets-1 contributes to destabilization of a quiescent EC phenotype in favor of an invasive angiogenic one, in part, by increasing expression of MMPs and integrin molecules that favor migration and invasion. Transfection of ECs with Ets-1 antisense prior to co-culture with E2 resulted in a 95% inhibition in capillary formation. We demonstrate here, for the first time that nanomolar concentrations of E2 directly and rapidly induced new capillary formation in a mammary tumor/EC co-culture system and suggest that this response may be mediated, in part, by an E2-induced increase in Ets-1 expression.

Vascular stroma-derived endothelial colony forming progenitor cells adapt to tumour growth.

Under appropriate nutrient agar culture conditions, primary or xenografted human and animal tumour biopsy-derived cell suspensions will form two types of colony. The first type, consisting of tight colonies of round cells which form tumours when introduced into nude mice, is of neoplastic origin. The second type of colony, the cells of which fail to form tumours on injection into nude mice, consists of loose colonies of larger, inter-connecting elongated bi- or tripolar cells and is thought to originate from vascular stroma-derived endothelial colony forming progenitor cells (V-ECPC). The likely importance of V-ECPC to tumour growth is emphasised by a positive correlation between the VECPC-derived endothelial cell colonies and both tumour vascularity and growth rate. A high cloning efficiency obtained from tumours of particularly intense vascular nature indicates that this cell is of importance in vascular adaptation and therefore tumour growth. In contrast, avascular, fibrotic tumour tissue yielded very low numbers of stromal vascular endothelial cell colonies. The results suggest that stromal vascular endothelial cell colonies do not arise from the mature fibroblastic elements of the tumour stroma, but rather from cells within actively growing regions. Tritiated thymidine uptake studies show that the vascular stroma-derived endothelial colony forming progenitor cells cell are cycling. Cell separation studies have characterized the as yet morphologically unidentified V-ECPC as having a sedimentation rate of 4.7 mm./hr and a mean density of 1.068 g/cm3 and hence a calculated volume of 450 microns 3.

Fertility cycle influence on surgical breast cancer cure.

Cancer growth and spread is an intricate process dependent upon both tumor and host. This laboratory is interested in the role of the fertility cycle, specifically cyclic changes in steroid hormone levels, in tumor growth and metastases. Our previous studies, using a murine model, have documented that breast cancer growth rate and post-resection metastatic behavior each change reproducibly during the estrous cycle, and that post-resection cancer spread depends upon the time within the estrous cycle that an advanced transplanted cancer is resected. Twelve to thiry-two percent cure rates were seen in these studies. That early work described estrous cycle stages just prior and near to putative ovulation to be superior while those stages farther from ovulation were disadvantageous times for surgery. Data presented here confirm the role of the estrous cycle in post-resection metastatic spread. This current work validates vaginal smear determined estrous cycle stage with uterine weight. A primary, transplantable, mammary carcinoma, which metastasizes to the lungs, was resected for surgical cure in cycling C3HeB/FeJ female mice at each fertility cycle stage. A group of oophorectomized (ovx) animals was also used. In two large, independent studies resecting much earlier stage cancers than in prior studies, a 96% surgical cure frequency was documented when the tumor is resected during estrus. The second best surgical cure rate is achieved when tumors are resected during metestrus (79% overall cure rate). Cure frequency in ovx animals is intermediate. These results further support a probable role for circulating E2 and P4 levels in modulating the metastatic process. We conclude that the timing of surgical resection within the estrous cycle affects the cancer's metastatic potential and that the optimal timing of resection may also depend to some extent upon the size (stage) of the resected cancer.

Effect of resveratrol on growth of 4T1 breast cancer cells in vitro and in vivo.

In vitro, resveratrol inhibited growth of 4T1 breast cancer cells in a dose- and time-dependent manner. In vivo, however, resveratrol had no effect on time to tumor take, tumor growth, or metastasis when administered intraperitoneally daily (1, 3, or 5 mg/kg) for 23 days starting at the time of tumor inoculation. Resveratrol had no effect on body weight, organ histology, or estrous cycling of the tumor-bearing mice. Resveratrol, therefore, is a potent inhibitor of 4T1 breast cancer cells in vitro; is nontoxic to mice at 1-5 mg/kg; and has no growth-inhibitory effect on 4T1 breast cancer in vivo.

THE EVOLUTION OF CARBON ALLOCATION TO PLANT SECONDARY METABOLITES: A GENETIC ANALYSIS OF COST IN DIPLACUS AURANTIACUS.

Diplacus aurantiacus contains large amounts of a leaf phenolic resin, an important deterrent to a leaf-eating caterpillar, Euphydryas chalcedona. The resin can also retard water loss during drought. Furthermore, the leaf resin content differs among plants and populations. This study investigates the existence of heritable variation (h2 ) in resin production and tests for a genetic correlation (rG ) between carbon allocation to secondary metabolites and growth rate, as well as with three other vegetative traits. Nine dam and 10 sire plants were chosen randomly at a field site and used to generate 78 full-sib families (19 half-sib families) by crossing all males to all females in a factorial design. Heritability was estimated in two ways, and genetic correlations were estimated by three methods. We found: (1) the heritability of resin production estimated by the regression of offspring on sires was significantly greater than zero (hs2=0.32, P<0.01); (2) the maternal variance in resin content was significantly greater than zero (21.3% of total phenotypic variance); (3) significant negative genetic correlation between resin content and growth rate was observed from two of three methods and was consistent with the phenotypic correlation; and (4) the cost of resin could be assessed quantitatively. The genetic cost of 1 mg in resin is equivalent to 25 mg of dry shoot-biomass growth, but the phenotypic cost is only 2.1 mg. This study indicates that carbon allocation to these secondary metabolites may respond to natural selection, and the phenotypic cost of resin production has a genetic basis in D. aurantiacus. This trade-off suggests that once selection occurs, increased phenolic resin production may result in decreased growth, or vice versa.