Barley ROP binding kinase1 is involved in microtubule organization and in basal penetration resistance to the barley powdery mildew fungus

Certain plant receptor-like cytoplasmic kinases were reported to interact with small monomeric G-proteins of the RHO of plant (ROP; also called RAC) family in planta and to be activated by this interaction in vitro. We identified a barley (Hordeum vulgare) partial cDNA of a ROP binding protein kinase (HvRBK1) in yeast (Saccharomyces cerevisiae) two-hybrid screenings with barley HvROP bait proteins. Protein interaction of the constitutively activated (CA) barley HvROPs CA HvRACB and CA HvRAC1 with full-length HvRBK1 was verified in yeast and in planta. Green fluorescent protein-tagged HvRBK1 appears in the cytoplasm and nucleoplasm, but CA HvRACB or CA HvRAC1 can recruit green fluorescent protein-HvRBK1 to the cell periphery. Barley HvRBK1 is an active kinase in vitro, and activity is enhanced by CA HvRACB or GTP-loaded HvRAC1. Hence, HvRBK1 might act downstream of active HvROPs. Transient-induced gene silencing of barley HvRBK1 supported penetration by the parasitic fungus Blumeria graminis f. sp. hordei, suggesting a function of the protein in basal disease resistance. Transient knockdown of HvRBK1 also influenced the stability of cortical microtubules in barley epidermal cells. Hence, HvRBK1 might function in basal resistance to powdery mildew by influencing microtubule organization.     

Continuous fluorescence microphotolysis and correlation spectroscopy using 4Pi microscopy

Continuous fluorescence microphotolysis (CFM) and fluorescence correlation spectroscopy (FCS) permit measurement of molecular mobility and association reactions in single living cells. CFM and FCS complement each other ideally and can be realized using identical equipment. So far, the spatial resolution of CFM and FCS was restricted by the resolution of the light microscope to the micrometer scale. However, cellular functions generally occur on the nanometer scale. Here, we develop the theoretical and computational framework for CFM and FCS experiments using 4Pi microscopy, which features an axial resolution of ∼100 nm. The framework, taking the actual 4Pi point spread function of the instrument into account, was validated by measurements on model systems, employing 4Pi conditions or normal confocal conditions together with either single- or two-photon excitation. In all cases experimental data could be well fitted by computed curves for expected diffusion coefficients, even when the signal/noise ratio was small due to the small number of fluorophores involved.     

Microarray Data Reveal Relationship between Jag1 and Ddr1 in Mouse Liver

Alagille syndrome is an autosomal dominant disorder involving bile duct paucity and cholestasis in addition to cardiac, skeletal, ophthalmologic, renal and vascular manifestations. Mutations in JAG1, encoding a ligand in the Notch signaling pathway, are found in 95% of patients meeting clinical criteria for Alagille syndrome. In order to define the role of Jag1 in the bile duct developmental abnormalities seen in ALGS, we previously created a Jag1 conditional knockout mouse model. Mice heterozygous for the Jag1 conditional and null alleles demonstrate abnormalities in postnatal bile duct growth and remodeling, with portal expansion and increased numbers of malformed bile ducts. In this study we report the results of microarray analysis and identify genes and pathways differentially expressed in the Jag1 conditional/null livers as compared with littermate controls. In the initial microarray analysis, we found that many of the genes up-regulated in the Jag1 conditional/null mutant livers were related to extracellular matrix (ECM) interactions, cell adhesion and cell migration. One of the most highly up-regulated genes was Ddr1, encoding a receptor tyrosine kinase (RTK) belonging to a large RTK family. We have found extensive co-localization of Jag1 and Ddr1 in bile ducts and blood vessels in postnatal liver. In addition, co-immunoprecipitation data provide evidence for a novel protein interaction between Jag1 and Ddr1. Further studies will be required to define the nature of this interaction and its functional consequences, which may have significant implications for bile duct remodeling and repair of liver injury.     

Expression of Multiple Sexual Signals by Fathers and Sons in the East-Mediterranean Barn Swallow: Are Advertising Strategies Heritable?

The level of expression of sexually selected traits is generally determined by genes, environment and their interaction. In species that use multiple sexual signals which may be costly to produce, investing in the expression of one sexual signal may limit the expression of the other, favoring the evolution of a strategy for resource allocation among signals. As a result, even when the expression of sexual signals is condition dependent, the relative level of expression of each signal may be heritable. We tested this hypothesis in the East-Mediterranean barn swallow (Hirundo rustica transitiva), in which males have been shown to express two uncorrelated sexual signals: red-brown ventral coloration, and long tail streamers. We show that variation in both signals may partially be explained by age, as well as by paternal origin (genetic father-son regressions), but that the strongest similarity between fathers and sons is the relative allocation towards one trait or the other (relative expression index), rather than the expression of the traits themselves. These results suggest that the expression of one signal is not independent of the other, and that genetic strategies for resource allocation among sexual signals may be selected for during the evolution of multiple sexual signals.     

In vivo and in vitro SAR of tetracyclic MAPKAP-K2 (MK2) inhibitors. Part II

Spirocyclopropane- and spiroazetidine-substituted tetracycles 13D–E and 16A are described as orally active MK2 inhibitors. The spiroazetidine derivatives are potent MK2 inhibitors with IC₅₀ <3 nM and inhibit the release of TNFα (IC₅₀<0.3 μM) from hPBMCs and hsp27 phosphorylation in anisomycin stimulated THP-1 cells. The spirocyclopropane analogues are less potent against MK2 (IC₅₀ = 0.05–0.23 μM), less potent in cells (IC₅₀ <1.1 μM), but show good oral absorption. Compound 13E (100 mg/kg po; bid) showed oral activity in rAIA and mCIA, with significant reduction of swelling and histological score.     

A Predominant Multidrug-Resistant Salmonella enterica Serovar Saintpaul Clonal Line in German Turkey and Related Food Products

Recently, Salmonella enterica subsp. enterica serovar Saintpaul has increasingly been observed in several countries, including Germany. However, the pathogenic potential and epidemiology of this serovar are not very well known. This study describes biological attributes of S. Saintpaul isolates obtained from turkeys in Germany based on characterization of their pheno- and genotypic properties. Fifty-five S. Saintpaul isolates from German turkeys and turkey-derived food products isolated from 2000 to 2007 were analyzed by using antimicrobial agent, organic solvent, and disinfectant susceptibility tests, isoelectric focusing, detection of resistance determinants, plasmid profiling, pulsed-field gel electrophoresis (PFGE), and hybridization experiments. These isolates were compared to an outgroup consisting of 24 S. Saintpaul isolates obtained from humans and chickens in Germany and from poultry and poultry products (including turkeys) in Netherlands. A common core resistance pattern was detected for 27 German turkey and turkey product isolates. This pattern included resistance (full or intermediate) to ampicillin, amoxicillin-clavulanic acid, gentamicin, kanamycin, nalidixic acid, streptomycin, spectinomycin, and sulfamethoxazole and intermediate resistance or decreased susceptibility to ciprofloxacin (MIC, 2 or 1 μg/ml, respectively) and several third-generation cephalosporins (including ceftiofur and cefoxitin [MIC, 4 to 2 and 16 to 2 μg/ml, respectively]). These isolates had the same core resistance genotype, with bla_TEM-1, aadB, aadA2, sul1, a Ser83→Glu83 mutation in the gyrA gene, and a chromosomal class 1 integron carrying the aadB-aadA2 gene cassette. Their XbaI, BlnI, and combined XbaI-BlnI PFGE patterns revealed levels of genetic similarity of 93, 75, and 90%, respectively. This study revealed that a multiresistant S. Saintpaul clonal line is widespread in turkeys and turkey products in Germany and was also detected among German human fecal and Dutch poultry isolates.Over the last few decades, the emergence and spread of antimicrobial agent-resistant zoonotic bacteria has become a serious public health concern (2, 23). The widespread use of antimicrobial agents for disease control, including at the farm level, has increased selection of antimicrobial agent-resistant Salmonella isolates (1, 23, 44). Food animals are considered an important reservoir for resistant bacteria. These animals and food products derived from them are traded worldwide, which contributes to the global spread of zoonotic agents and antimicrobial resistance. In the last few years, several monitoring activities were initiated in order to generate baseline data on antimicrobial resistance in bacteria isolated from livestock and food derived from animals that could be used in future assessments of the risk of antimicrobial resistance (10).According to European Union (EU) Zoonoses Regulation (EC) no. 2160/2003 on the control of Salmonella and other specified food-borne zoonotic agents, a European Community target for reducing the prevalence of Salmonella in turkey flocks had to be established. Consequently, EU Commission decision 2006/662/EC was released, and a baseline survey of the prevalence of Salmonella in turkey flocks was carried out in all European countries, including Germany, over a 1-year period starting on 1 October 2006 (http://www.efsa.europa.eu/EFSA/efsa_locale-1178620753812_1178706574172.htm). The main objective of this study was to estimate the prevalence of Salmonella in commercial flocks of turkeys. The data showed that at the EU level Salmonella enterica serovar Bredeney was the serovar reported most frequently for fattening turkey flocks and occurred in 17.2% of the samples from Salmonella-positive flocks (1,084 of 3,702 flocks were positive), followed by S. enterica serovar Hadar, S. enterica serovar Derby, and then S. enterica serovar Saintpaul (14.0%, 11.3%, and 10.4% of the samples from positive flocks, respectively). In this study, S. Saintpaul was detected in fattening turkeys in 12 countries, reflecting the wide spread of this serovar. Recently, S. Saintpaul has been increasingly observed in several countries, including Germany. According to Enter-Net reports (data on Salmonella human isolates identified by European national reference centers), for the last quarter of the year 2006 S. Saintpaul was the fourth most common serovar (1.6%) and, in contrast to the data for previous years, was one of the most frequent causes of human salmonellosis in Europe. After this, its prevalence was 1.2% and 0.6% in the first quarters of 2007 and 2008, respectively, among the Salmonella serotypes implicated in human disease (http://ecdc.europa.eu/en/publications/Pages/Surveillance_Reports.aspx). During the period from 2001 to 2009 in Germany, 463 cases of human salmonellosis related to S. Saintpaul (0.09% of all cases; the maximum prevalence was 0.15% in 2008, the prevalence was 0.1% in 2002, 2005, 2006, and 2009 and 0.06% in 2004, and the minimum prevalence was 0.05% in 2007) were reported to the Robert Koch Institute (Berlin, Germany) (www3.rki.de/SurvStat). In Germany, S. Saintpaul attracted public attention particularly in 1993, when it caused a nationwide food-borne outbreak (27). This serotype has often been related to outbreaks in other countries, and in 2008 it was implicated in a large multistate human outbreak associated with various vegetables in the United States (4).Previous studies showed that isolates of S. Saintpaul are often multidrug resistant (33, 35), but little is known about the mechanisms underlying antimicrobial resistance or about the pathogenic potential and epidemiology of isolates belonging to this serotype. The goals of this study were to obtain information about the resistance characteristics of isolates collected between 2000 and 2007 in Germany and to assess possible clonal relationships. The isolates used originated from turkey feces collected during the German Salmonella baseline study (in 2006 and 2007) or from diagnostic samples, including samples of turkey feces and turkey-related food products. These isolates were compared with German strains isolated from humans and chickens and with poultry strains isolated in Netherlands.     

Tumor necrosis factor and norepinephrine lower the levels of human neutrophil peptides 1-3 secretion by mixed synovial tissue cultures in osteoarthritis and rheumatoid arthritis

Introduction  

Neutrophils and monocytes play an important role in overt inflammation in chronic inflammatory joint diseases such as rheumatoid arthritis (RA). The sympathetic nervous system (SNS) inhibits many neutrophil/monocyte functions and macrophage tumor necrosis factor (TNF), but because of the loss of sympathetic nerve fibers in inflamed tissue, sympathetic control is attenuated. In this study, we focused on noradrenergic and TNF regulation of human neutrophil peptides 1-3 (HNP1-3), which are proinflammatory bactericidal α-defensins.     

Abnormal calcium metabolism in normocalcaemic sarcoidosis.

In studies of calcium metabolism in 13 unselected patients with untreated sarcoidosis all were normocalcaemic but five had hypercalcuria. All had normal renal function. Calcium absorption was indexed by a double isotope test. 45Ca hyperabsorption occurred in six patients. Ten kinetic studies were carried out with 47Ca and in six bone turnover was increased. 45Ca absorption correlated well with the calculated bone uptake rate of calcium, and with urine calcium excretion. These results suggest that in sarcoidosis abnormalities in calcium metabolism are fairly common although they rarely result in sustained hypercalcaemia.