bluuebunny/crossref_metadata_2025 huggingface dataset:
Rhizobium leguminosarum biovar viciae 3841, deficient in 27-hydroxyoctacosanoate-modified lipopolysaccharide, is impaired in desiccation tolerance, biofilm formation and motility
Authors: Vanderlinde Elizabeth M. , Muszyński Artur , Harrison Joe J. , Koval Susan F. , Foreman Dallas L. , Ceri Howard , Kannenberg Elmar L. , Carlson Russell W. , Yost Christopher K.
Published: August 2009

The lipopolysaccharide (LPS) of the Gram-negative legume symbiontRhizobium leguminosarumbiovarviciae3841 contains several unique modifications, including the addition of a 27-hydroxyoctacosanoic acid (27OHC28 : 0), also termed the very long chain fatty acid (VLCFA), attached at the 2′ position of lipid A. A transposon mutant that lacks expression of two putative 3-oxo-acyl [acyl-carrier protein] synthase II genes,fabF1andfabF2, from the VLCFA biosynthetic cluster, was isolated and characterized. MS indicated that the lipid A of the mutant lacked the VLCFA modification, and sodium deoxycholate (DOC)-PAGE of the LPS indicated further structural alterations. The mutant was characteristically sensitive to several stresses that would be experienced in the soil environment, such as desiccation and osmotic stresses. An increase in the excretion of neutral surface polysaccharideswas observed in the mutant. This mutant was also altered in its attachment to solid surfaces, and was non-motile, with most of the mutant cells lacking flagella. Despite the pleiotropic effects of the mutation, these mutants were still able to nodulate legumes and fix atmospheric nitrogen. This report emphasizes that a structurally intact VLCFA-containing lipid A is critical to cellular traits that are important for survival in the rhizosphere.
Rhizobium etli HrpW is a pectin-degrading enzyme and differs from phytopathogenic homologues in enzymically crucial tryptophan and glycine residues
Authors: Fauvart Maarten , Verstraeten Natalie , Dombrecht Bruno , Venmans Ruth , Beullens Serge , Heusdens Christophe , Michiels Jan
Published: August 2009

While establishing a nitrogen-fixing symbiosis with leguminous plants, rhizobia are faced with the problem of penetrating the plant cell wall at several stages of the infection process. One of the major components of this barrier is pectin, a heteropolysaccharide composed mainly of galacturonic acid subunits. So far, no enzymes capable of degrading pectin have been isolated from rhizobia. Here, we make an inventory of rhizobial candidate pectinolytic enzymes based on available genome sequence data and present an initial biochemical and functional characterization of a protein selected from this list. Rhizobium etli hrpW is associated with genes encoding a type III secretion system, a macromolecular structure that allows bacteria to directly inject so-called effector proteins into a eukaryotic host's cell cytosol and an essential virulence determinant of many Gram-negative pathogenic bacteria. In contrast to harpin HrpW from phytopathogens, R. etli HrpW possesses pectate lyase activity and is most active on highly methylated substrates. Through comparative sequence analysis, three amino acid residues crucial for the observed enzymic activity were identified: Trp192, Gly212 and Gly213. Their importance was confirmed by site-directed mutagenesis and biochemical characterization of the resulting proteins, with the tryptophan mutant showing no detectable pectate lyase activity and the double-glycine mutant's activity reduced by about 80 %. Surprisingly, despite hrpW expression being induced specifically on the plant root surface, a knockout mutation of the gene does not appear to affect symbiosis with the common bean Phaseolus vulgaris.
The transcription factor DNR from Pseudomonas aeruginosa specifically requires nitric oxide and haem for the activation of a target promoter in Escherichia coli
Authors: Castiglione Nicoletta , Rinaldo Serena , Giardina Giorgio , Cutruzzolà Francesca
Published: August 2009

Pseudomonas aeruginosa is a well-known pathogen in chronic respiratory diseases such as cystic fibrosis. Infectivity of P. aeruginosa is related to the ability to grow under oxygen-limited conditions using the anaerobic metabolism of denitrification, in which nitrate is reduced to dinitrogen via nitric oxide (NO). Denitrification is activated by a cascade of redox-sensitive transcription factors, among which is the DNR regulator, sensitive to nitrogen oxides. To gain further insight into the mechanism of NO-sensing by DNR, we have developed an Escherichia coli-based reporter system to investigate different aspects of DNR activity. In E. coli DNR responds to NO, as shown by its ability to transactivate the P. aeruginosa
norCB promoter. The direct binding of DNR to the target DNA is required, since mutations in the helix–turn–helix domain of DNR and specific nucleotide substitutions in the consensus sequence of the norCB promoter abolish the transcriptional activity. Using an E. coli strain deficient in haem biosynthesis, we have also confirmed that haem is required in vivo for the NO-dependent DNR activity, in agreement with the property of DNR to bind haem in vitro. Finally, we have shown, we believe for the first time, that DNR is able to discriminate in vivo between different diatomic signal molecules, NO and CO, both ligands of the reduced haem iron in vitro, suggesting that DNR responds specifically to NO.
The xrvA gene of Xanthomonas oryzae pv. oryzae, encoding an H-NS-like protein, regulates virulence in rice
Authors: Feng Jia-Xun , Song Zhi-Zhong , Duan Cheng-Jie , Zhao Shuai , Wu Ying-Qiao , Wang Chao , Dow J. Maxwell , Tang Ji-Liang
Published: August 2009

Xanthomonas oryzaepv.oryzae(Xoo) causes bacterial blight disease in rice, one of the most serious rice diseases. ThexrvAgene fromXoostrain 13751 encodes a protein containing a histone-like nucleoid-structuring protein (H-NS) domain. The expression ofxrvAin strain 13751 was enhanced in XOM2 minimal medium. Mutation of thexrvAgene of strain 13751 led to a significant reduction in virulence in the host plant rice, a delayed hypersensitive response in the nonhost castor-oil plant, a decrease in extracellular polysaccharide and diffusible signal factor production, and an increase in intracellular glycogen accumulation. Northern hybridization analyses revealed that the virulence-associated geneshrpG,hrpX,rpfC,rpfF,rpfGandgumBwere downregulated in thexrvAmutant compared to the wild-type and complemented strains. Interestingly, increase of copy number ofxrvAin the wild-type strain 13751 resulted in a strain showing similar phenotypes as thexrvAmutant and a reduction of the expression ofgumB,hrpX,rpfC,rpfFandrpfG. These findings indicate that thexrvAgene, which is highly conserved in the sequenced strains ofXanthomonas, encodes an important regulatory factor for the virulence ofXoo.
Quorum sensing differentially regulates Pseudomonas aeruginosa type VI secretion locus I and homologous loci II and III, which are required for pathogenesis
Authors: Lesic B. , Starkey M. , He J. , Hazan R. , Rahme L. G.
Published: August 2009

Pseudomonas aeruginosa harbours three type VI secretion (T6S) loci. Although HSI-I has been partially studied, limited knowledge is available on the homologous loci HSI-II and HSI-III. We show that quorum sensing (QS) differentially regulates the expression of genes at all three loci. HSI-I-associated gene expression is suppressed by both the homoserine lactone transcription factor LasR and the 4-hydroxy-2-alkylquinoline (HAQ) transcriptional regulator MvfR. Conversely, both HSI-II and HSI-III loci are positively controlled by LasR and MvfR. PqsE, a key component of the MvfR regulon, is required for the expression of part of HSI-III but not HSI-II, and previously identified inhibitors of HAQ biosynthesis significantly downregulate HSI-II and -III gene expression. Animal and plant infection studies reveal that both HSI-II and -III play important roles in pathogenesis. Furthermore, analysis of a double ΔHSI-II : : III mutant suggests that these loci functionally compensate for one another in virulence. This study illustrates the contribution of the QS systems to T6S gene regulation and reveals the importance of HSI-II and -III in mediating P. aeruginosa pathogenesis. Moreover, this work provides new insights into the design and development of selective compounds that may restrict human P. aeruginosa and possibly other clinical infections.
Uptake of the fluorescent probe FM4-64 by hyphae and haemolymph-derived in vivo hyphal bodies of the entomopathogenic fungus Beauveria bassiana
Authors: Lewis Michael W. , Robalino Ines V. , Keyhani Nemat O.
Published: August 2009

The entomopathogenic fungus Beauveria bassiana is under intensive study as a pest biological control agent. B. bassiana produces several distinct single-cell types that include aerial conidia, in vitro blastospores and submerged conidia. Under appropriate nutrient conditions these cells can elaborate germ tubes that form hyphae, which in turn lead to the formation of a fungal mycelium. In addition, B. bassiana displays a dimorphic transition, producing in vivo specific yeast-like hyphal bodies during growth in the arthropod haemolymph. The amphiphilic styryl dye FM4-64 was used to investigate internalization and morphological features of in vitro and in vivo insect haemolymph-derived B. bassiana cells. In vitro blastospores and submerged conidia displayed a punctate pattern of internal labelling, whereas aerial conidia failed to internalize the dye under the conditions tested. FM4-64 was also taken up into both apical and subapical compartments of living hyphae in a time-dependent manner, with clearly observable vesicle labelling. Internalization, where occurring, was reversibly disrupted by lowering the temperature of the assay or by treatment with azide/fluoride and latrunculin A. Treatment with cytochalasin D and monensin also caused abnormal vesicle trafficking, although some staining of vesicles was noted. Fungal cells derived from infected Heliothis virescens haemolymph (in vivo cells) actively internalized FM4-64. The in vivo blastospores or hyphal bodies displayed bright membrane and internal vesicle staining, although diffuse staining of internal structures was also visible. These results suggest active uptake by different developmental stages of B. bassiana, including haemolymph-derived cells that can evade the insect immune system.
DNA polymerase X from Deinococcus radiodurans implicated in bacterial tolerance to DNA damage is characterized as a short patch base excision repair polymerase
Authors: Khairnar Nivedita P. , Misra Hari S.
Published: August 2009

TheDeinococcus radioduransR1 genome encodes an X-family DNA repair polymerase homologous to eukaryotic DNA polymeraseβ. The recombinant deinococcal polymerase X (PolX) purified from transgenicEscherichia colishowed deoxynucleotidyltransferase activity. Unlike the Klenow fragment ofE. coli, this enzyme showed short patch DNA synthesis activity on heteropolymeric DNA substrate. The recombinant enzyme showed 5′-deoxyribose phosphate (5′-dRP) lyase activity and base excision repair functionin vitro,with the help of externally supplied glycosylase and AP endonuclease functions. ApolXdisruption mutant ofD. radioduransexpressing 5′-dRP lyase and a truncated polymerase domain was comparatively less sensitive toγ-radiation than apolXdeletion mutant. Both mutants showed higher sensitivity to hydrogen peroxide. Excision repair mutants ofE. coliexpressing this polymerase showed functional complementation of UV sensitivity. These results suggest the involvement of deinococcal polymerase X in DNA-damage tolerance ofD. radiodurans, possibly by contributing to DNA double-strand break repair and base excision repair.
Inactivation of the ilvB1 gene in Mycobacterium tuberculosis leads to branched-chain amino acid auxotrophy and attenuation of virulence in mice
Authors: Awasthy Disha , Gaonkar Sheshagiri , Shandil R. K. , Yadav Reena , Bharath Sowmya , Marcel Nimi , Subbulakshmi Venkita , Sharma Umender
Published: August 2009

Acetohydroxyacid synthase (AHAS) is the first enzyme in the branched-chain amino acid biosynthesis pathway in bacteria. Bioinformatics analysis revealed that theMycobacterium tuberculosisgenome contains four genes (ilvB1,ilvB2,ilvGandilvX) coding for the large catalytic subunit of AHAS, whereas only one gene (ilvN or ilvH) coding for the smaller regulatory subunit of this enzyme was found. In order to understand the physiological role of AHAS in survival of the organismin vitroandin vivo, we inactivated theilvB1gene ofM. tuberculosis. The mutant strain was found to be auxotrophic for all of the three branched-chain amino acids (isoleucine, leucine and valine), when grown with either C6or C2carbon sources, suggesting that theilvB1gene product is the major AHAS inM. tuberculosis. Depletion of these branched chain amino acids in the medium led to loss of viability of the ΔilvB1strainin vitro, resulting in a 4-log reduction in colony-forming units after 10 days. Survival kinetics of the mutant strain cultured in macrophages maintained with sub-optimal concentrations of the branched-chain amino acids did not show any loss of viability, indicating either that the intracellular environment was rich in these amino acids or that the other AHAS catalytic subunits were functional under these conditions. Furthermore, the growth kinetics of the ΔilvB1strain in mice indicated that although this mutant strain showed defective growthin vivo, it could persist in the infected mice for a long time, and therefore could be a potential vaccine candidate.
Multiple redundant stress resistance mechanisms are induced in Salmonella enterica serovar Typhimurium in response to alteration of the intracellular environment via TLR4 signalling
Authors: Wright John A. , Tötemeyer Sabine S. , Hautefort Isabelle , Appia-Ayme Corinne , Alston Mark , Danino Vittoria , Paterson Gavin K. , Mastroeni Pietro , Ménager Nathalie , Rolfe Matthew , Thompson Arthur , Ugrinovic Sanja , Sait Leanne , Humphrey Tom , Northen Helen , Peters Sarah E. , Maskell Duncan J. , Hinton Jay C. D. , Bryant Clare E.
Published: August 2009

Toll-like receptor 4 (TLR4) senses bacterial LPS and is required for the control of systemicSalmonella entericaserovar Typhimurium infection in mice. The mechanisms of TLR4 activation and its downstream signalling cascades are well described, yet the direct effects on the pathogen of signalling via this receptor remain unknown. To investigate this we used microarray-based transcriptome profiling of intracellularS. Typhimurium during infection of primary bone marrow-derived macrophages from wild-type and TLR4-deficient mice. We identified 17S.Typhimurium genes that were upregulated in the presence of functional TLR4. Nine of these genes have putative functions in oxidative stress resistance. We therefore examinedS. Typhimurium gene expression during infection of NADPH oxidase-deficient macrophages, which lack normal oxidative killing mechanisms. We identified significant overlap between the ‘TLR4-responsive’ and ‘NADPH oxidase-responsive’ genes. This is new evidence for a link between TLR4 signalling and NADPH oxidase activity. Interestingly, with the exception of adpsmutant,S.Typhimurium strains lacking individual TLR4- and/or oxidative stress-responsive genes were not attenuated during intravenous murine infections. Our study shows that TLR4 activity, either directly or indirectly, induces the expression of multiple stress resistance genes during the intracellular life ofS.Typhimurium.
Biodegradation of phenanthrene by Pseudomonas sp. strain PPD: purification and characterization of 1-hydroxy-2-naphthoic acid dioxygenase
Authors: Deveryshetty Jaigeeth , Phale Prashant S.
Published: August 2009

Pseudomonassp. strain PPD can metabolize phenanthrene as the sole source of carbon and energy via the ‘phthalic acid’ route. The key enzyme, 1-hydroxy-2-naphthoic acid dioxygenase (1-HNDO, EC 1.13.11.38), was purified to homogeneity using a 3-hydroxy-2-naphthoic acid (3-H2NA)-affinity matrix. The enzyme was a homotetramer with a native molecular mass of 160 kDa and subunit molecular mass of ∼39 kDa. It required Fe(II) as the cofactor and was specific for 1-hydroxy-2-naphthoic acid (1-H2NA), withKm13.5 μM andVmax114 μmol min−1 mg−1. 1-HNDO failed to show activity with gentisic acid, salicylic acid and other hydroxynaphthoic acids tested. Interestingly, the enzyme showed substrate inhibition with aKiof 116 μM. 1-HNDO was found to be competitively inhibited by 3-H2NA with aKiof 24 μM. Based on the pH-dependent spectral changes, the enzyme reaction product was identified as 2-carboxybenzalpyruvic acid. Under anaerobic conditions, the enzyme failed to convert 1-H2NA to 2-carboxybenzalpyruvic acid. Stoichiometric studies showed the incorporation of 1 mol O2into the substrate to yield 1 mol product. These results suggest that 1-HNDO fromPseudomonassp. strain PPD is an extradiol-type ring-cleaving dioxygenase.
Smad‐3 as a mediator of the fibrotic response
Authors: Flanders Kathleen , Sato Misako , Ooshima Akira , Russo Angelo , Roberts Anita
Published: September 2009

Introduction  Smad‐3, a key cytoplasmic mediator of transforming growth factor‐β (TGF‐β) signalling, mediates many of its inflammatory and fibrotic effects in vivo (Roberts et al. 2001). Smad‐3 null mice are protected against cutaneous injury induced by ionizing irradiation (Flanders et al. 2002). Here, we report on our continuing studies on radioprotection as well as protection against tubulointerstitial fibrosis following unilateral ureteral obstruction (UUO) in Smad‐3 null mice.

Methods  For radioprotection studies, the flank skin of Smad‐3+/+ wild‐type (WT) and Smad‐3–/– knockout (KO) mice was exposed to 30 Gy of localized γ‐irradiation and analysed for histology and gene expression at various times post irradiation. In the UUO model, the right proximal ureter of WT and KO mice was ligated, and 1–2 weeks later kidneys were analysed for inflammation, fibrosis and gene expression.

Results  Six weeks after exposure to irradiation, skin from KO mice shows less epidermal acanthosis and influx of mast cells, macrophages and neutrophils than skin of WT mice. Paradoxically, at 6–8 h post irradiation, KO skin shows a significantly greater number of neutrophils. Irradiated KO skin also exhibits less immunoreactive TGF‐β, fewer myofibroblasts and less scarring than does WT. Smad‐3 null dermal fibroblasts do not respond to the chemotactic effects of TGF‐β and show less induction of fibrogenic cytokines when treated with irradiation plus TGF‐β compared to WT cells. Following UUO, normal kidney architecture is preserved in KO mice, while kidneys from WT mice are enlarged with an influx of mononuclear cells and increased expression of collagen and TGF‐β1. Additionally, renal tubules in obstructed kidneys of KO mice remain positive for E‐cadherin without expression of α‐smooth muscle actin, while the opposite expression pattern is seen in obstructed kidneys of WT mice. TGF‐β treatment of primary cultures of WT renal tubular epithelial cells results in a phenotypic change from a cobblestone pattern to a spindle‐shaped fibroblastic appearance, while KO cells treated with TGF‐β maintain their original appearance.

Conclusion  Smad‐3 plays an important role in mediating pathogenic inflammation and fibrosis in several model systems and is also essential for TGF‐β1‐induced epithelial–mesenchymal transition in renal tubular epithelial cells. Inhibitors of the Smad‐3 pathway may have clinical applications in the treatment of a number of fibrotic conditions.
Identification of ribosomal protein L19 as a novel tumor antigen recognized by autologous cytotoxic T lymphocytes in lung adenocarcinoma
Authors: Kuroda Koji , Takenoyama Mitsuhiro , Baba Tetsuro , Shigematsu Yoshiki , Shiota Hironobu , Ichiki Yoshinobu , Yasuda Manabu , Uramoto Hidetaka , Hanagiri Takeshi , Yasumoto Kosei
Published: September 2009

The purpose of the present study was to identify a novel tumor‐specific antigen capable of inducing a specific cellular immune response in lung cancer patients. The co‐culture of regional lymph node lymphocytes and the CD80‐transfected autologous lung adenocarcinoma cell line H1224L resulted in a successful induction of bulk cytotoxic T lymphocytes (CTL). CTL clone L7/8 was established by the limiting dilution method from these bulk CTLs and lysed H1224L but not autologous Epstein–Barr virus‐transformed B cells or K562. The CTL clone also recognized allogeneic lung cancer cell lines in an HLA‐A*31012‐restricted manner. Using the CTL clone, an antigen‐coding gene was identified using the cDNA expression cloning technique, which encodes ribosomal protein L19 (RPL19). Finally, a 9 mer antigenic peptide was identified by means of construction of mini‐genes. RPL19 was overexpressed in the lung cancer tissue from patient H1224. All of the normal tissues examined expressed lower levels of RPL19 mRNA than that of the lung cancer tissue. RPL19 was also found to be overexpressed in 12 of 30 (40%) non‐small‐cell lung cancer tissues by immunohistochemical staining. The expression level of RPL19 in tumor cell lines correlated positively with the production of interferon (IFN)‐γby CTL clone L7/8 in response to such cell lines. In addition, the suppression of RPL19 expression by transfection with small interfering RNA resulted in the suppression of cyclinD1, D3 synthesis, and the growth inhibition of lung cancer cell lines overexpressing RPL19. Therefore, this growth suppression could be ascribed to the inhibition of the cell cycle. These results may indicate that RPL19 is a novel overexpressed antigen which may therefore be a useful candidate as a target for specific immunotherapy. (Cancer Sci 2009)
A comparison of sex ratio, birth periods and calf survival among Serengeti wildebeest sub‐populations, Tanzania
Authors: Ndibalema Vedasto G.
Published: August 2009

Although adaptation and environmental conditions can easily predict demographic variation in most savannah ungulates, no study on demographic consequences arising from natural and anthropogenic factors among Serengeti wildebeest (Connochaetes taurinus) sub‐populations in Tanzania has been carried out. Here, I report estimates of annual sex ratio, calf and yearling survival rate and birth seasonality between resident and migratory sub‐populations to explore demographic patterns arising from the different age and sex structure. The results indicate significantly higher female‐biased sex ratios in the resident and almost even sex ratios among individual migrants. The calf recruitment estimated as mother: offspring ratios indicate a more synchronous birth in the migrant than the resident sub‐population. Also, birth seasonality in the migratory sub‐population coincided with seasonal variability of rainfall and the timing of the birth peak was more variable in the migrants than the resident sub‐population. The migratory sub‐population had a higher annual proportional mean calf survival estimate (0.84) than that of the residents (0.44) probably due to higher mortality resulting from predation in the western corridor. However, the proportion of yearling survival estimates was much lower (0.31) in the migrants and relatively higher (0.39) in the residents. Different demographic outcomes resulting from environment, predation, movements and ecological factors including resource competition have conservation implications for the two sub‐populations.
Effect of eland density and foraging onCombretum apiculatumphysiognomy in a semi‐arid savannah
Authors: Nyengera Reason , Sebata Allan
Published: August 2009

We studied the impacts of eland density on the physiognomy ofCombretum apiculatum. We divided the study area into two zones: high eland density zone (≤8 km from watering point) and low eland density zone (≥8 km from watering point). Eland density was determined in each zone using walking transects.Combretum apiculatumheight and diameter and levels of eland damage were measured in each zone. Eland density was 1.92 and 0.86 km−2for high and low‐density zones respectively. We recorded 239C. apiculatumtrees, 138 in low‐density zone and 101 in high‐density zone. MeanC. apiculatumheight was 2.88 ± 0.67 m and 5.05 ± 1.04 m for high and low eland density zones respectively. High eland density prevented the recruitment ofC. apiculatumfrom the 2.6–5.5 m height class to the >5.6 m height class. Although extensive tree damage occurred at <2.5 m height stratum,C. apiculatumshowed resilience as recruitment into the 2.6–5.5 m height class continued. We concluded that high eland density prevents recruitment ofC. apiculatumto higher height classes while at the same time causing extensive tree damage at lower height strata.
The feeding choice of the Red Fox (Vulpes vulpes) in a semi‐arid fragmented landscape of North Africa in relation to water and energy contents of prey
Authors: Dell’Arte Graziella L. , Leonardi Giovanni
Published: August 2009

In semi‐arid zones, the lack of surface water for drinking forced predators to obtain it from the body fluids of prey and this requires more different prey than that needed to provide energy alone. We examined Red fox (Vulpes vulpes) scats to ascertain if selective feeding compensates the shortage of free water by the ingestion of more prey rich in water such as insects. In addition, we discussed our results in light of increasing habitat fragmentation that, in turn, influences the prey availability. At different spatial levels, recombination of prey assemblages in the diet reaches a solution for variable shortage of water. This was true especially at the landscape level (different degree of aridity) and indirectly for habitat types. Nevertheless, the effects of landscape characteristics probably influenced foxes through the influence of suitable habitats, which partitioned prey species. In fact, increasing values of patchiness had a negative effect on measured energetic gain indices but foxes try to exploiting different patches including small and isolated ones because these surveys become important to obtain water moistures from alternative prey (Orthopters).
The anti‐leukaemic activity of novel synthetic naphthoquinones against acute myeloid leukaemia: induction of cell death via the triggering of multiple signalling pathways
Authors: Hallak Maher , Win Thida , Shpilberg Ofer , Bittner Shmuel , Granot Yosef , Levy Itai , Nathan Ilana
Published: August 2009

Naphthoquinones, such as menadione, display lower toxicity than anthracyclins used in cancer chemotherapy. Novel anti‐leukaemic compounds comprised of chloro‐amino‐phenyl naphthoquinones with substitutions on the benzoic ring were developed. Structure–activity relationship studies indicated that the analogue with both methyl and amine substitutions (named TW‐92) was the most efficient in killing leukaemic cells. Treatment of U‐937 promonocytic cells with TW‐92 induced apoptotic or necrotic cell death, dependent on incubation and dose conditions. TW‐92 induced rapid phosphorylation of p38 mitogen‐activated protein kinase (p38MAPK) and of extracellular signal‐regulated protein kinases (ERK1/2). The generation of apoptosis was preceded by intracellular H2O2 accumulation accompanied by glutathione depletion, the former inhibited by di‐phenyl‐iodonium (DPI), an inhibitor of NADPH oxidase. TW‐92 induced swelling of isolated rat liver mitochondria, indicative of a direct effect on mitochondria. Apoptosis in intact cells was accompanied by a decrease in mitochondrial membrane potential, cytochrome c release and caspase activation. In addition, the level of Mcl‐1, an anti‐apoptotic regulatory protein, was down‐regulated, whereas the expression of the pro‐apoptotic BAX was elevated. Finally, TW‐92 exerted strong pro‐apoptotic and necrotic effects in primary acute myeloid leukaemia samples when given in submicromolar concentrations. Together, these findings demonstrate that TW‐92 may provide an effective anti‐leukaemic strategy.
The utility of volunteer home‐visiting support to prevent maternal depression in the first year of life
Authors: Barnes J. , Senior R. , MacPherson K.
Published: August 2009

Background  Maternal depression can be detrimental to infant development. Structured home visiting initiated either in pregnancy or soon after the birth by a professional has led to better outcomes for mothers and their children but some vulnerable families may respond more favourably to a local volunteer. The value of volunteer support provided in the UK by Home‐Start for maternal well‐being is noted in qualitative studies, but there is no evidence of its impact from trials. The support is not structured and both the frequency and content of visits may vary.

Methods  A cluster randomized study allocated Home‐Start local schemes to intervention or control conditions. Mothers in all areas were screened at routine health checks in late pregnancy. In intervention areas names of those scoring 9+ on the Social Disadvantage Screening Index were passed to Home‐Start to be offered a volunteer. Not all those offered the support accepted the offer. In control areas no support was offered. Research assessments were conducted at 2 and 12 months. The outcomes were major or minor depression occurring between 2 and 12 months (Structured Clinical Interview for Diagnostic and Statistical Manual – Third Edition – Revised) and depression symptoms at 12 months (Edinburgh Postnatal Depression Scale). Three groups were compared: supported, case‐matched controls and those offered but not receiving support.

Results  Almost one‐third experienced depression during the time period. Volunteer support had no identifiable impact on the emergence of maternal depression from 2 to 12 months or on depression symptoms when infants were 12 months. The major predictor of both was depression identified at 2 months.

Conclusions  It was not found that informal support initiated following screening for disadvantage in pregnancy reduced the likelihood of depression for mothers with infants.
Profiling of factor VIII mutations in Korean haemophilia A
Authors: HWANG S. H. , KIM M. J. , LIM J. A. , KIM H. C. , KIM H. S.
Published: August 2009

Summary.  Haemophilia A (HA) is an X chromosome‐linked inherited bleeding disorder caused by heterogeneous mutations of coagulation factor VIII (FVIII). Although more than 900 mutations of FVIII gene are reported in the HAMSTeRS database, the mutation data regarding the FVIII gene in the Korean population is currently insufficient. The aim of this study was to profile the mutations of FVIII in Korean HA, 38 unrelated Korean HA male patients were examined. Peripheral blood samples were obtained from the patients. Long distance‐PCR was performed for the identification of inversions in intron 22 and intron 1. Then gross exon deletion was examined to the inversion‐negative patients by multiplex‐PCR. Finally, direct sequencing was performed on exons 1–26, 5’‐ and 3’‐UTR. We identified 33 mutations from the 38 patients. These included 15 inversions in intron 22 (39.5%), one inversion in intron 1 (2.6%), one gross exon deletion (2.6%), five deletions (13.2%), two insertions (5.3%), six missense (15.8%) and three nonsense mutations (7.9%). Mutation types for five patients (13.2%) were not identified in this study. We determined that the most common defect in FVIII in this study was an inversion mutation in intron 22; this is consistent with the findings of other studies. For the first time in Korean HA, a patient with intron 1 inversion was found. In addition, we report eight novel mutation types which never been reported in HAMSTeRS database. The mutation data in this study should prove useful as a reference for the diagnosis of HA and the detection of carriers in the Korean population.
Medical exposures in youth and the frequency of narcolepsy with cataplexy: a population‐based case–control study in genetically predisposed people
Authors: KOEPSELL THOMAS D. , LONGSTRETH WILLIAM T. , TON THANH G. N.
Published: August 2009

Epidemiological observations suggest that exposures in youth may trigger narcolepsy in genetically predisposed individuals. In this population‐based case–control study, we sought to identify all prevalent cases of narcolepsy with cataplexy aged 18–50 years as of 1 July 2001, in King County, Washington. The 45 eligible cases who were DQB1*0602‐positive were compared with 95 controls with this allele, identified through random‐digit dialing and buccal smears. Cases and controls were interviewed in person about physician‐diagnosed infectious and non‐infectious illnesses, immunizations, head trauma and parasomnias or psychiatric problems during youth. Narcolepsy with cataplexy was more frequent in African‐Americans and in poorer households. Adjusting for these factors, the condition was 5.4‐fold more common [95% confidence interval (CI) = 1.5–19.1] among people reporting a physician‐diagnosed strep throat before the age of 21 years. No other significant associations with childhood diseases, immunizations or head trauma were found. However, prevalence was increased 16.3‐fold (95% CI = 6.1–44.1) in subjects who reported having had ‘night terrors’. Strep throat may be related to narcolepsy with cataplexy in genetically susceptible individuals. The association with night terrors could simply reflect early symptoms of narcolepsy, or they could be a prodromal sign of disturbed sleep physiology.
Sleep problems in general practice: a national survey of assessment and treatment routines of general practitioners in Norway
Authors: SIVERTSEN BØRGE , NORDHUS INGER H. , BJORVATN BJØRN , PALLESEN STÅLE
Published: August 2009

The aim of the current national survey of all general practitioners (GPs) practising in Norway was to explore assessment, treatment practice and perceived efficacy of treatment of sleep problems in general practice. A short questionnaire, including self‐report measures of the frequency and routines of treatment and assessment of sleep problems, was sent to all 4049 GPs in Norway, of whom 1465 (36.7%) provided valid responses. The prevalence of sleep problems among patients in general practice was estimated to be 11.2%, of which almost two‐thirds were believed to be caused by a medical condition. Anamnestic information and blood tests were the most common assessment of sleep problems, whereas GPs rarely referred patients to all‐night polysomnographic recording. Sleep hygiene advices were the most commonly used treatment strategy, whereas hypnotics were believed to have the best short‐term efficacy. Antidepressives were considered to be the best option for long‐term management of sleep problems. About one‐third of the patients were prescribed benzodiazepines or ‘Z‐drugs’ for more than 6 months. This study demonstrates that sleep problems are recognized by GPs, but despite evidence that non‐pharmacological treatment is superior in the long‐term management of insomnia, the current study shows that hypnotics are still considered by GPs to be the most successful treatment.
Comparative study of temperature‐dependent life histories of three economically important Adelphocoris spp.
Authors: LU YANHUI , WU KONGMING , WYCKHUYS KRISA G. , GUO YUYUAN
Published: August 2009

Subsequent to the widespread adoption of Bt transgenic cotton in China and an associated reduction in pesticide use, Adelphocoris spp. (Hemiptera: Miridae) are the key pests of this crop. Three species (Adelphocoris suturalis, Adelphocoris fasciaticollis and Adelphocoris lineolatus) are found in Chinese Bt cotton fields, each with a distinct geographic distribution and phenology. In the present study, the development and fecundity of the three species are compared in the laboratory at various temperatures in the range 10–35 °C. Although nymphal development and adult moulting occurs under all temperature regimes, egg eclosion is not observed at 10 °C. In general, egg and nymphal development periods decrease with increasing temperature up to 30 °C. The lower and upper development thresholds are, respectively, 5.6 and 40.1 °C for A. suturalis eggs; 5.0 and 38.4 °C for nymphs; 6.3 and 39.0 °C for A. fasciaticollis eggs, 3.0 and 41.9 °C for nymphs; 5.6 and 41.3 °C for A. lineolatus eggs; and 6.2 and 38.8 °C for nymphs. Thermal constants are 189.9 degree days (DD) (egg) and 308.8 DD (nymph) for A. suturalis, 188.8 DD (egg) and 366.7 DD (nymph) for A. fasciaticollis, and 231.7 DD (egg) and 291.6 DD (nymph) for A. lineolatus. Temperatures above 30 °C affect egg development of A. fasciaticollis and A. lineolatus adversely, but not that of A. suturalis. At the same time, nymphal survival of A. suturalis is reduced at 10 °C. Longevity of all species declines with increasing temperature, whereas extremes of temperature (i.e. 10 and 35 °C) interfere with oviposition. The estimated optimum range for oviposition is 23–25 °C, irrespective of species. In general, development and fecundity of the three Adelphocoris spp. is consistent with their respective distribution and seasonal dynamics. The present study provides insight into the distribution and phenology of Adelphocoris spp., and contributes to the modelling of their population dynamics.
Determination of effective light pulse and classical Bünsow experiment in the larval diapause of the Indian meal moth Plodia interpunctella
Authors: KIKUKAWA SHIGERU , HIGUCHI AKANE , IKEDA TATSUYA , MUKAI TAKESHI , OKAMOTO HIROYUKI , TANAKA KOUJI , YAGUCHI SYUHEI
Published: August 2009

Plodia interpunctella Hübner (Lepidoptera: Pyralidae) comprises a model insect to analyse the photoperiodic time‐measuring system controlling its larval diapause. In the present study, the effective length of light pulse in night interruption experiments is determined at 25 °C. Various lengths of light pulse are tested by inserting them at the midnight of an LD 12 : 12 h photoperiod. When the light pulse is 15 or 30 min, the incidence of diapause is 86%. To inhibit the induction of diapause effectively, a light pulse of 1.75–2 h is needed. The incidence of diapause is 12% under an LD 12 : 5 : 2 : 5 h photoperiod. To determine the precise role of the light pulse, 2‐h light pulses placed at the midnight of an LD 12 : 12 h photoperiod are disrupted systematically by darkness. When a 2‐h light pulse is disrupted by 15 min of darkness, diapause is generally prevented (< 29%) regardless of the temporal position of darkness. Longer disruption by darkness induces diapause moderately (37–67%). A Bünsow experiment is also conducted at 25 and 20 °C, in which the main photophase of 12 h of light is combined with 24–72‐h scotophases scanned by a 2‐h light pulse. The photoperiodic cycle length tested, therefore, varies in the range 36–84 h. In each cycle length, the incidence of diapause fluctuates as the light pulse moves toward dawn. However, no regular and circadian changes in the percentage diapause are observed in relation to diapause determination.
Proteomic analysis ofβ‐aminobutyric acid priming and abscisic acid – induction of drought resistance in crabapple (Malus pumila): effect on general metabolism, the phenylpropanoid pathway and cell wall enzymes
Authors: MACARISIN DUMITRU , WISNIEWSKI MICHAEL E. , BASSETT CAROLE , THANNHAUSER THEODORE W.
Published: August 2009

In a variety of herbaceous, model and crop plants, DL‐β‐aminobutyric acid (BABA), has been shown to enhance both biotic and abiotic stress resistance by potentiating rather than inducing resistance responses but studies in woody plants are lacking. In the present study, two‐dimensional difference in‐gel electrophoresis (DIGE) was used to quantify differences in protein abundance in leaf tissue from BABA‐treated, abscisic acid (ABA)‐treated and untreated (control) plants during a 10 d drought stress. ABA‐treated seedlings were most resistant to water loss followed by BABA‐treated seedlings and then untreated seedlings. Although some similarity was observed in the proteome of ABA‐ and BABA‐treated seedlings, a dramatic shift in the proteome occurred earlier in ABA‐treated seedlings. Some proteins showed almost identical patterns of increase or decrease in abundance in both BABA‐ and ABA‐treated seedlings, supporting the concept that BABA‐induced abiotic stress resistance in plants is achieved by potentiating an ABA‐regulated pathway. Some proteins, however, were induced or suppressed only in BABA‐primed plants, indicating that BABA may also mediate resistance via some ABA‐independent pathways. Based on the putative function of the identified proteins, we propose that changes in cell wall enzymes and a suppression of lignin biosynthesis may play a specific role in BABA‐primed drought resistance.
Probiotics for allergic respiratory diseases – Putting it into perspective
Authors: Singh Meenu , Ranjan Das Rashmi
Published: August 2009

Singh M, Das RR. Probiotics for allergic respiratory diseases – Putting it into perspective.  Pediatr Allergy Immunol 2010: 21: e368–e376. © 2009 John Wiley & Sons A/S

Respiratory allergies include allergic rhinitis, sinusitis and asthma. Increasing attention on pathogenesis of allergic airway diseases has given rise to “atopic march” hypothesis i.e. clinical features of atopic eczema occur first and precede the development of asthma and allergic rhinitis. The “hygiene hypothesis” proposes that the increase in allergic diseases reflects a decrease in infections during childhood. Clinical trials also suggest that the exposure to microbes through the gastrointestinal tract powerfully shapes immune function. Probiotics are live organisms which exert a beneficial effect in the prevention as well as treatment of allergic diseases through modification of immune system of host via gut ecosystem. Intestinal microbiota differs in infants who later develop allergic diseases, and feeding probiotics to infants at risk has been shown to reduce their rate of developing eczema. This has prompted studies of feeding probiotics in prevention as well as treatment of respiratory allergy. We hereby discuss the status of probiotics in respiratory allergy.
Relationship between infant weight gain and later asthma
Authors: Paul Ian M. , Camera Lindsay , Zeiger Robert S. , Guilbert Theresa W. , Bacharier Leonard B. , Taussig Lynn M. , Morgan Wayne J. , Covar Ronina A. , Krawiec Marzena , Bloomberg Gordon R. , Mauger David T. ,   for the Childhood Asthma Research and Education (CARE) Network
Published: August 2009

Paul IM, Camera L, Zeiger RS, Guilbert TW, Bacharier LB, Taussig LM, Morgan WJ, Covar RA, Krawiec M, Bloomberg GR, Mauger DT, for the Childhood Asthma Research and Education (CARE) Network. Relationship between infant weight gain and later asthma. Pediatr Allergy Immunol 2010: 21: 82–89. © 2009 John Wiley & Sons A/S

Like obesity, the prevalence of asthma has increased over the past several decades. Accelerated patterns of infant growth have been associated with obesity and its co‐morbidities. We aimed to determine if infant weight gain pattern is associated with asthma development later in childhood. Birth weight, growth, pulmonary function, and symptom data were collected in a trial of 2‐ to 3‐yr‐old children at‐risk for asthma randomized to a 2‐yr treatment with inhaled corticosteroids or placebo followed by a 1‐yr observation period of study medication. Patterns of infant weight gain between birth and study enrollment were categorized as accelerated, average, or decelerated. Regression analyses were used to test the effects of infant weight gain pattern prior to study enrollment on outcomes during the observation year and at study conclusion while adjusting for demographics, baseline symptom severity, study treatment, and atopic indicators. Among the 197 study participants, early life weight gain pattern was not associated with daily asthma symptoms or lung function at the study’s conclusion. However, both prednisone courses (p = 0.01) and urgent physician visits (p < 0.001) were significantly associated with weight gain pattern with fewer exacerbations occurring amongst those with a decelerated weight gain pattern. We conclude that early life patterns of weight change were associated with subsequent asthma exacerbations, but were not associated with asthma symptoms or pulmonary function during the pre‐school years for these children at‐risk for asthma.
Implementing a pre‐operative checklist to increase patient safety: a 1‐year follow‐up of personnel attitudes
Authors: NILSSON L. , LINDBERGET O. , GUPTA A. , VEGFORS M.
Published: August 2009

Background: The operating room is a complex work environment with a high potential for adverse events. Protocols for perioperative verification processes have increasingly been recommended by professional organizations during the last few years. We assessed personnel attitudes to a pre‐operative checklist (‘time out’) immediately before start of the operative procedure.

Methods: ‘Time out’ was implemented in December 2007 as an additional safety barrier in two Swedish hospitals. One year later, in order to assess how the checklist was perceived, a questionnaire was sent by e‐mail to 704 persons in the operating departments, including surgeons, anesthesiologists, operation and anesthetic nurses and nurse assistants. In order to identify differences in response between professions, each alternative in the questionnaire was assigned a numerical value.

Results: The questionnaire was answered by 331 (47%) persons and 93% responded that ‘time out’ contributes to increased patient safety. Eighty‐six percent thought that ‘time out’ gave an opportunity to identify and solve problems. Confirmation of patient identity, correct procedure, correct side and checking of allergies or contagious diseases were considered ‘very important’ by 78–84% of the responders. Attitudes to checking of patient positioning, allergies and review of potential critical moments were positive but differed significantly between the professions. Attitudes to a similar checklist at the end of surgery were positive and 72–99% agreed to the different elements.

Conclusion: Staff attitudes toward a surgical checklist were mostly positive 1 year after their introduction in two large hospitals in central Sweden.
Single nucleotide polymorphisms of 17β‐hydroxysteroid dehydrogenase type 7 gene: Mechanism of estramustine‐related adverse reactions?
Authors: Ozeki Takeshi , Takeuchi Megumi , Suzuki Motofumi , Kitamura Tadaichi , Takayanagi Risa , Yokoyama Haruko , Yamada Yasuhiko
Published: September 2009

Objectives:  To investigate the influence of single nucleotide polymorphisms (SNP) on transcription of the 17β‐hydroxysteroid dehydrogenase (HSD17B7) gene.

Methods:  Luciferase reporter genes containing a 5′‐flanking of the HSD17B7 gene, as well as the sequence around the SNP, were transfected into LNCaP and DU145 cells. Then, luciferase assays were carried out.

Results:  The presence of the G allele resulted in an increase of transcriptional activity derived from the 5′‐flanking region of the HSD17B7 gene by 270% and 370% in LNCaP and DU145 cells, respectively. Transcriptional activity of the HSD17B7 gene containing the G allele was higher than that of the C allele.

Conclusions:  The transcriptional activity of the HSD17B7 gene containing the G allele is higher than that of the C allele. This difference in HSD17B7 expression may regulate the risk of peripheral edema as an adverse reaction induced by estramustine phosphate sodium.
Modifying techniques in deep inferior epigastric artery perforator flap harvest with the use of preoperative imaging
Authors: Rozen Warren M. , Ashton Mark W.
Published: August 2009

New techniques in the harvest of deep inferior epigastric artery perforator (DIEP) flaps have become introduced as a result of modern imaging technologies that can allow virtual surgery to be achieved preoperatively. With computed tomographic angiography, individual anatomy can be appreciated in detail to a level not previously appreciated. These imaging techniques can be successfully used to guide DIEP flap surgery. ‘Optimal’ perforators can be selected based on size, location, intramuscular and subcutaneous course, and their association with motor nerves. Flap design can be safely achieved based on the cutaneous distribution of perforators. Abdominal wall closure can be improved based on the abdominal contour seen with imaging. Preoperative planning can aid patient selection, plan all aspects of the operative technique, reduce operating time and improve operative outcomes.