bluuebunny/crossref_metadata_2025 huggingface dataset:
Antagonism of Trichoderma harzianum ETS 323 on Botrytis cinerea Mycelium in Culture Conditions
Authors: Cheng Chi-Hua , Yang Chia-Ann , Peng Kou-Cheng
Published: June 2012

Previous studies have shown that the extracellular proteins of Trichoderma harzianum ETS 323 grown in the presence of deactivated Botrytis cinerea in culture include a putative l-amino acid oxidase and have suggested the involvement of this enzyme in the antagonistic mechanism. Here, we hypothesized that the mycoparasitic process of Trichoderma spp. against B. cinerea involves two steps; that is, an initial hyphal coiling stage and a subsequent hyphal coiling stage, with different coiling rates. The two-step antagonism of T. harzianum ETS 323 against B. cinerea during the mycoparasitic process in culture was evaluated using a biexponential equation. In addition, an l-amino acid oxidase (Th-l-AAO) was identified from T. harzianum ETS 323. The secretion of Th-l-AAO was increased when T. harzianum ETS 323 was grown with deactivated hyphae of B. cinerea. Moreover, in vitro assays indicated that Th-l-AAO effectively inhibited B. cinerea hyphal growth, caused cytosolic vacuolization in the hyphae, and led to hyphal lysis. Th-l-AAO also showed disease control against the development of B. cinerea on postharvest apple fruit and tobacco leaves. Furthermore, an apoptosis-like response, including the generation of reactive oxygen species, was observed in B. cinerea after treatment with Th-l-AAO, suggesting that Th-l-AAO triggers programmed cell death in B. cinerea. This may be associated with the two-step antagonism of T. harzianum ETS 323 against B. cinerea.
Factors Affecting Admission to Anesthesiology Residency in the United States
Authors: de Oliveira Gildàsio S. , Akikwala Tulsi , Kendall Mark C. , Fitzgerald Paul C. , Sullivan John T. , Zell Christopher , McCarthy Robert J.
Published: June 2012

Background

Admission to an anesthesiology residency in the United States is competitive, and the odds associated with a successful match based on the applicants' characteristics have not been determined. The objective of this study was to examine factors associated with admission to anesthesiology residency in the United States.

Methods

The study was a retrospective cohort evaluation of the 2010 to 2011 residency applicants. Applicants' characteristics and objective factors used to select trainees were extracted. The primary outcome was a successful match to an anesthesiology residency. Data were analyzed using conditional inference tree analysis and propensity score matching.

Results

Data available from 1,976 applications were examined corresponding to 58% of the national sample. The odds (99% CI) for successful match were 3.6 (3.1-4.2) for U.S. medical school graduates, 2.6 (2.3 to 3.0) for applicants with United States Medical Licensing Examination Step 2 scores more than 210, and 1.2 (1.1 to 1.3) for female applicants. The odds (99% CI) for a successful match for international and U.S. graduate applicants younger than 29 yr was 3.3 (2.0-5.4) and (1.9 to 4.2), respectively, even after propensity matching for medical school, exam scores, and gender. The average applicant had no peer-reviewed scholarly productivity.

Conclusion

Although anesthesiology residency acceptance was primarily associated with U.S. medical school attendance and United States Medical Licensing Examination Step 2 scores, our study suggest an influence of age and gender bias in the selection process. Peer-reviewed scholarly production among applicants and prior graduate education did not appear to influence candidate selection.
Cutaneous Vasoconstriction Affects Near-infrared Spectroscopy Determined Cerebral Oxygen Saturation during Administration of Norepinephrine
Authors: Sørensen Henrik , Secher Niels H. , Siebenmann Christoph , Nielsen Henning B. , Kohl-Bareis Matthias , Lundby Carsten , Rasmussen Peter
Published: June 2012

Background

Perioperative optimization of spatially resolved near-infrared spectroscopy determined cerebral frontal lobe oxygenation (scO2) may reduce postoperative morbidity. Norepinephrine is routinely administered to maintain cerebral perfusion pressure and, thereby, cerebral blood flow, but norepinephrine reduces the scO2. We hypothesized that norepinephrine-induced reduction in scO2 is influenced by cutaneous vasoconstriction.

Methods

Fifteen healthy male subjects (25 ± 5 yr, mean ± SD) were studied during: hyperventilation (1.5 kPa end-tidal PcO2 reduction), whole-body heating, administration of norepinephrine (0.15 μg · kg · min; with and without end-tidal carbon dioxide correction), and hypoxia (FiO2: 0.12%). Arterial (saO2), skin, and internal jugular venous oxygen saturations (sjO2) were recorded, and the average cerebral capillary oxygen saturation (scapO2) was calculated.

Results

This study indicates that scO2 is influenced by skin oxygen saturation because whole-body heating increased scO2 by 3.6% (2.1-5.1%; 95% CI) and skin oxygen saturation by 3.1% (1.3-4.9%), whereas scapO2 remained unaffected. Conversely, hyperventilation decreased scO2 by 2.1% (0.4-3.7%) and scapO2 by 5.3% (3.8-6.9%), whereas skin oxygen saturation increased 1.8% (0.5-3.1%). In response to hypoxia, scO2 (10.2%; 6.6-13.7%), scapO2 (7.9%; 6.4-9.4%), and skin oxygen saturation (8.9%; 6.3-11.6%) all decreased. With administration of norepinephrine there was a 2.2% (1.0-4.3%) decrease in skin oxygen saturation and scO2 decreased 6.2% (4.2-8.0%), with scapO2 remaining unaffected.

Conclusion

The results confirm that spatially resolved near-infrared spectroscopy detects cerebral deoxygenation with systemic hypoxic exposure and hyperventilation. However, a commonly used vasopressor norepinephrine disturbs skin oxygen saturation to an extent that influences scO2.
Volatile Anesthetics Reduce Invasion of Colorectal Cancer Cells through Down-regulation of Matrix Metalloproteinase-9
Authors: Müller-Edenborn Björn , Roth-Z'graggen Birgit , Bartnicka Kamila , Borgeat Alain , Hoos Alexandra , Borsig Lubor , Beck-Schimmer Beatrice
Published: June 2012

Background

Invasion of extracellular matrix is a hallmark of malignant tumors. Clamping maneuvers during cancer surgery reduce blood loss, but trigger reperfusion injury (RI). RI increases cancer recurrence in the reperfused organ through up-regulation of matrix metalloproteinase-9 (MMP-9). Interleukin-8 is an important cytokine in RI promoting accumulation of neutrophils, a major source of MMP-9. Volatile anesthetics were demonstrated to reduce RI. We hypothesized that these anesthetics might attenuate MMP-9 up-regulation and consequently tumor cell invasion in RI.

Methods

Isolated human neutrophils (n = 6) were preconditioned with sevoflurane or desflurane, followed by stimulation with interleukin-8, phorbol myristate acetate, or chemokine CXC-ligand 1 (CXCL1) to differentiate intracellular pathways. MMP-9 release and activity were quantified by enzyme-linked immunosorbent assay and zymography, respectively. CXC-receptor-2 (CXCR2) expression and phosphorylation of extracellular signal-regulated kinases 1/2 were assessed by flow cytometry. The impact of MMP-9 on the invasion of neutrophils and MC-38 colon cancer cells was assessed using Matrigel-coated filters (n = 6).

Results

Preconditioning reduced interleukin-8-induced MMP-9-release by 41% (±13, 5%, sevoflurane) and 40% (±13%, desflurane). This was also evident following stimulation of CXCR2 with CXCL1. No impact on phosphorylation of extracellular signal-regulated kinases 1/2 and MMP-9 release was observed with receptor-independent stimulation of protein kinase C with phorbol myristate acetate. Preconditioning reduced transmigration of neutrophils and MC-38 tumor cells to baseline levels.

Discussion

Volatile anesthetics impair neutrophil MMP-9 release and interfere with pathways downstream of CXCR2, but upstream of protein kinase C. Through down-regulation of MMP-9, volatile anesthetics decrease Matrigel breakdown and reduce subsequent migration of cancer cells in vitro.
In Vitro Hemostatic Properties of French Lyophilized Plasma
Authors: Martinaud Christophe , Civadier Corinne , Ausset Sylvain , Verret Catherine , Deshayes Anne-Virginie , Sailliol Anne
Published: June 2012

Background

French lyophilized plasma (FLyP) is used routinely by the French Armed Forces in war settings. The authors compared concentrations of coagulation proteins and global in vitro hemostatic properties in FLyP and in the same plasma before lyophilization to assess the impact of lyophilization on coagulation properties.

Methods

Twenty-four batches of plasma before and after lyophilization were tested for coagulation proteins. Thrombin generation time, thrombin antithrombin concentration, prothrombin fragment 1 + 2, and thromboelastography were assessed. Finally, the efficiencies of FLyP and plasma before lyophilization were compared on a hemorrhagic shock hemodilution model and tested on TEG(Haemoscope Corporation, Glenview, IL).

Results

Prothrombin time ratio (1.1 ± 0.1 vs. 1.2 ± 0.1) and activated partial thromboplastin time (35 ± 1.3 vs. 39 ± 2.4 s) were significantly increased in FLyP (8 ± 3%, P < 0.05 and 11 ± 5%, P < 0.001, respectively). Activity of factors V (85 ± 18 vs. 51 ± 16 UI/ml) and VIII (0.77 ± 0.11 vs. 0.62 ± 0.10 UI/ml) was also diminished (25 ± 12% and 20 ± 7%, respectively); however, activity of other factors was preserved. The authors observed no alteration in the thromboelastographic parameters. Thrombin generation was preserved when induced with 5 pM tissue factor in vitro but significantly reduced when using 1 pM tissue factor. The thrombin-antithrombin complex and prothrombin fragment 1 + 2 attested for the absence of coagulation activation. This hemodilution model showed no significant difference before and after lyophilization.

Conclusions

The study results account for a significant decrease of factors V and VIII in FLyP. However, the global capacity to induce clot formation in vitro seems to be preserved. The clinical relevance of these decreased factors is not known.
Differential Effects of Peripheral versus Central Coadministration of QX-314 and Capsaicin on Neuropathic Pain in Rats
Authors: Shen Jun , Fox Lyle E. , Cheng Jianguo
Published: June 2012

Background

Neuropathic pain is common and difficult to treat. Recently a technique was developed to selectively inhibit nociceptive inputs by simultaneously applying two drugs: capsaicin, a transient receptor potential vanilloid receptor-1 channel activator, and QX-314, a lidocaine derivative that intracellularly blocks sodium channels. We used this technique to investigate whether transient receptor potential vanilloid receptor 1-expressing nociceptors contribute to neuropathic pain.

Methods

The rat chronic constriction injury model was used to induce neuropathic pain in order to test the analgesic effects of both peripheral (perisciatic) and central (intrathecal) administration of the QX-314/capsaicin combination. The Hargreaves and von Frey tests were used to monitor evoked pain-like behaviors and visual observations were used to rank spontaneous pain-like behaviors.

Results

Perisciatic injections of the QX-314/capsaicin combination transiently increased the withdrawal thresholds by approximately 3-fold, for mechanical and thermal stimuli in rats (n = 6/group) with nerve injuries suggesting that peripheral transient receptor potential vanilloid receptor 1-expressing nociceptors contribute to neuropathic pain. In contrast, intrathecal administration of the QX-314/capsaicin combination did not alleviate pain-like behaviors (n = 5/group). Surprisingly, intrathecal QX-314 alone (n = 9) or in combination with capsaicin (n = 8) evoked spontaneous pain-like behaviors.

Conclusions

Data from the perisciatic injections suggested that a component of neuropathic pain was mediated by peripheral nociceptive inputs. The role of central nociceptive terminals could not be determined because of the severe side effects of the intrathecal drug combination. We concluded that only peripheral blockade of transient receptor potential vanilloid receptor 1-expressing nociceptive afferents by the QX-314/capsaicin combination was effective at reducing neuropathic allodynia and hyperalgesia.
Antiinflammation Effect of Human Placental Multipotent Mesenchymal Stromal Cells Is Mediated by Prostaglandin E2 via a Myeloid Differentiation Primary Response Gene 88-dependent Pathway
Authors: Chen Chie-Pein , Tsai Pei-Shan , Huang Chun-Jen
Published: June 2012

Background

We sought to elucidate the antiinflammation effect of human placental multipotent mesenchymal stromal cells (hPMSCs) and the possible molecular mechanisms.

Methods

Immortalized murine macrophages (RAW264.7 cells), with or without hPMSCs coincubation, were treated with endotoxin to induce expression of the relevant molecules.

Results

The peak concentrations (means ± SD) of inflammatory molecules in endotoxin-activated macrophages with hPMSCs coincubation were significantly lower than those in endotoxin-activated macrophages without hPMSCs coincubation (tumor necrosis factor-α: 9.4±0.8 vs. 13.0±1.1 ng/ml; interleukin-6: 0.8±0.1 vs. 1.2±0.1 ng/ml; macrophage inflammatory protein-2: 345±30 vs. 666±51 ng/ml; intercellular adhesion molecule 1: 1.4±0.1 vs. 1.7±0.1 ng/ml; prostaglandin E2: 5.7±0.3 vs. 8.5±0.6 ng/ml; all P<0.008). Data of the activation of nuclear factor-κB and mitogen-activated protein kinases as well as the interaction between toll-like receptor 4 and myeloid differentiation primary response gene 88 paralleled those of the inflammatory molecules. In contrast, the endotoxin binding and toll-like receptor 4/myeloid differential-2 complex activation in endotoxin-activated macrophages with hPMSCs coincubation were comparable with those in endotoxin-activated macrophages without hPMSCs coincubation. As our data revealed that hPMSCs could induce low-grade prostaglandin E2 expression in macrophages, we also employed the selective cyclooxygenase-2 inhibitor NS-398 to further elucidate the possible role of prostaglandin E2. Our data revealed that the above-mentioned hPMSC-triggered inhibitory effects were significantly reversed by NS-398.

Conclusions

The antiinflammation effect of human placental multipotent mesenchymal stromal cells is mediated, at least in part, by prostaglandin E2 via a myeloid differentiation primary response gene 88-dependent pathway.
Superoscillation phenomena in SO(3)
Authors: Aharonov Y. , Colombo F. , Nussinov S. , Sabadini I. , Struppa D. C. , Tollaksen J.
Published: June 2012

We prove a new asymptotic formula for the Wigner
d
-functions, as a consequence of the discovery of superoscillatory behaviour in SO(3).
Existence of multiple vortices in supersymmetric gauge field theory
Authors: Chen Shouxin , Yang Yisong
Published: June 2012

Two sharp existence and uniqueness theorems are presented for solutions of multiple vortices arising in a six-dimensional brane-world supersymmetric gauge field theory under the general gauge symmetry group
G
=
U
(1)×
SU
(
N
) and with
N
Higgs scalar fields in the fundamental representation of
G
. Specifically, when the space of extra dimension is compact so that vortices are hosted in a 2-torus of volume |
Ω
|, the existence of a unique multiple vortex solution representing
n
1
,…,
n

N

, respectively, prescribed vortices arising in the
N
species of the Higgs fields is established under the explicitly stated necessary and sufficient condition

where
e
and
g
are the
U
(1) electromagnetic and
SU
(
N
) chromatic coupling constants,
v
measures the energy scale of broken symmetry and

is the total vortex number; when the space of extra dimension is the full plane, the existence and uniqueness of an arbitrarily prescribed
n
-vortex solution of finite energy is always ensured. These vortices are governed by a system of nonlinear elliptic equations, which may be reformulated to allow a variational structure. Proofs of existence are then developed using the methods of calculus of variations.
The Light-Response BTB1 and BTB2 Proteins Assemble Nuclear Ubiquitin Ligases That Modify Phytochrome B and D Signaling in Arabidopsis
Authors: Christians Matthew J. , Gingerich Derek J. , Hua Zhihua , Lauer Timothy D. , Vierstra Richard D.
Published: June 2012

Members of the Bric-a-Brac/Tramtrack/Broad Complex (BTB) family direct the selective ubiquitylation of proteins following their assembly into Cullin3-based ubiquitin ligases. Here, we describe a subfamily of nucleus-localized BTB proteins encoded by the LIGHT-RESPONSE BTB1 (LRB1) and LRB2 loci in Arabidopsis (Arabidopsis thaliana) that strongly influences photomorphogenesis. Whereas single lrb1 and lrb2 mutants are relatively normal phenotypically, double mutants are markedly hypersensitive to red light, but not to far-red or blue light, and are compromised in multiple photomorphogenic processes, including seed germination, cotyledon opening and expansion, chlorophyll accumulation, shade avoidance, and flowering time. This red light hypersensitivity can be overcome by eliminating phytochrome B (phyB) and phyD, indicating that LRB1/2 act downstream of these two photoreceptor isoforms. Levels of phyB/D proteins but not their messenger RNAs are abnormally high in light-grown lrb1 lrb2 plants, implying that their light-dependent turnover is substantially dampened. Whereas other red light-hypersensitive mutants accumulate phyA protein similar to or higher than the wild type in light, the lrb1 lrb2 mutants accumulate less, suggesting that LRB1/2 also positively regulate phyA levels in a phyB/D-dependent manner. Together, these data show that the BTB ubiquitin ligases assembled with LRB1/2 function redundantly as negative regulators of photomorphogenesis, possibly by influencing the turnover of phyB/D.
Purification, crystallization and preliminary crystallographic analysis of histone lysine demethylase NO66 fromHomo sapiens
Authors: Zhou Xing , Tao Yue , Wu Minhao , Zhang Dandan , Zang Jianye
Published: June 2012

NO66 is a JmjC domain-containing histone demethylase with specificity towards histone H3 methylated on both Lys4 and Lys36in vitroandin vivo. A fragment of NO66 lacking the N-terminal 167 amino-acid residues was overexpressed inEscherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method. X-ray diffraction data were collected to a resolution of 2.29 Å. NO66 crystallized in space groupP31orP32, with unit-cell parametersa= 89.35,b = 89.35,c= 304.86 Å, α = β = 90, γ = 120°, and the crystal is likely to contain four molecules in the asymmetric unit.
Crystallization of an atypical short-chain dehydrogenase fromVibrio vulnificuslacking the conserved catalytic tetrad
Authors: Buysschaert Geraldine , Verstraete Kenneth , Savvides Savvas N. , Vergauwen Bjorn
Published: June 2012

Short-chain dehydrogenases/reductases (SDRs) are a rapidly expanding superfamily of enzymes that are found in all kingdoms of life. Hallmarked by a highly conserved Asn-Ser-Tyr-Lys catalytic tetrad, SDRs have a broad substrate spectrum and play diverse roles in key metabolic processes. Locus tag VVA1599 inVibrio vulnificusencodes a short-chain dehydrogenase (hereafter referred to as SDRvv) which lacks the signature catalytic tetrad of SDR members. Structure-based protein sequence alignments have suggested that SDRvv may harbour a unique binding site for its nicotinamide cofactor. To date, structural studies of SDRs with altered catalytic centres are underrepresented in the scientific literature, thus limiting understanding of their spectrum of substrate and cofactor preferences. Here, the expression, purification and crystallization of recombinant SDRvv are presented. Two well diffracting crystal forms could be obtained by cocrystallization in the presence of the reduced form of the phosphorylated nicotinamide cofactor NADPH. The collected data were of sufficient quality for successful structure determination by molecular replacement and subsequent refinement. This work sets the stage for deriving the identity of the natural substrate of SDRvv and the structure–function landscape of typical and atypical SDRs.
Crystallization and preliminary X-ray crystallographic analysis of Pz peptidase B fromGeobacillus collagenovoransMO-1
Authors: Nakano Hiroaki , Hosokawa Allin , Tagawa Ryuji , Inaka Koji , Ohta Kazunori , Nakatsu Toru , Kato Hiroaki , Watanabe Kunihiko
Published: June 2012

Pz peptidase B is an intracellular M3 metallopeptidase that is found together with Pz peptidase A in the thermophileGeobacillus collagenovoransMO-1 and recognizes collagen-specific tripeptide units (-Gly-Pro-X-). These peptidases have low homology in their primary structures; however, their cleavage patterns towards peptide substrates are similar. In this work, Pz peptidase B was crystallized using the counter-diffusion method. Data were collected to a resolution of 1.6 Å at 100 K from a crystal obtained in the Japanese Experiment Module (JEM; also known as `Kibo') at the International Space Station (ISS). The crystal belonged to the trigonal space groupP3121, with unit-cell parametersa=b= 87.64,c= 210.5 Å. A complete data set was also obtained from crystals of selenomethionine-substituted protein.
Purification, crystallization and preliminary X-ray diffraction analysis of a hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT) fromCoffea canephorainvolved in chlorogenic acid biosynthesis
Authors: Lallemand Laura A. , McCarthy James G. , McSweeney Sean , McCarthy Andrew A.
Published: June 2012

Chlorogenic acids (CGAs) are a group of soluble phenolic compounds that are produced by a variety of plants, includingCoffea canephora(robusta coffee). The last step in CGA biosynthesis is generally catalysed by a specific hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (HQT), but it can also be catalysed by the more widely distributed hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT). Here, the cloning and overexpression of HCT fromC. canephorainEscherichia colias well as its purification and crystallization are presented. Crystals were obtained by the sitting-drop technique at 293 K and X-ray diffraction data were collected on the microfocus beamline ID23-2 at the ESRF. The HCT crystals diffracted to better than 3.0 Å resolution, belonged to space groupP42212 with unit-cell parametersa=b= 116.1,c= 158.9 Å and contained two molecules in the asymmetric unit. The structure was solved by molecular replacement and is currently under refinement. Such structural data are needed to decipher the molecular basis of the substrate specifities of this key enzyme, which belongs to the large plant acyl-CoA-dependent BAHD acyltransferase superfamily.
Expression, purification, crystallization and preliminary crystallographic analysis of the phosphoglycerate kinase fromAcinetobacter baumannii
Authors: Baretta Kayla , Garen Craig , Yin Jiang , James Michael N. G.
Published: June 2012

Acinetobacter baumanniiis a common multidrug-resistant clinical pathogen that is often found in hospitals. TheA. baumanniiphosphoglycerate kinase (AbPGK) is involved in the key energy-producing pathway of glycolysis and presents a potential target for antibiotic development.AbPGK has been expressed and purified; it was crystallized using lithium sulfate as the precipitant. TheAbPGK crystals belonged to space groupP2221. They diffracted to a resolution of 2.5 Å using synchrotron radiation at the Canadian Light Source.
Crystallization and preliminary X-ray diffraction analysis of the high molecular weight ketoacyl reductase FabG4 complexed with NADH
Authors: Dutta Debajyoti , Bhattacharyya Sudipta , Das Amit Kumar
Published: June 2012

FabG4 fromMycobacterium tuberculosisbelongs to the high molecular weight ketoacyl reductases (HMwFabGs). The enzyme requires NADH for β-ketoacyl reductase activity. The protein was overexpressed, purified to homogeneity and crystallized as a FabG4–NADH complex. A mountable FabG4:NADH complex crystal diffracted to 2.59 Å resolution and belonged to space groupP1, with unit-cell parametersa= 63.07,b= 71.03,c= 92.92 Å, α = 105.02, β = 97.06, γ = 93.66°. The Matthews coefficient suggested the presence of four monomers in the unit cell. In addition, a self-rotation function revealed the presence of two twofold NCS axes and one fourfold NCS axis. At χ = 180° the highest peak corresponds to the twofold NCS between two monomers, whereas the second peak corresponds to the twofold NCS between two dimers.
Purification, crystallization and preliminary X-ray diffraction analysis of the effector protein PevD1 fromVerticillium dahliae
Authors: Han Lei , Liu Zheng , Liu Xinqi , Qiu Dewen
Published: June 2012

The effector protein PevD1 from the pathogenic fungusVerticillium dahliaewas purified and crystallized using the hanging-drop vapour-diffusion method. Native crystals appeared in a solution consisting of 4.0 Msodium formate. A native data set was collected at 1.9 Å resolution at 100 K using an in-house X-ray source. Because of the absence of useful methinione in the protein sequence, derivative crystals that contained iodine were obtained by soaking in 1.25 Mpotassium iodide, and a data set that contained anomalous signal was collected using the same X-ray facility at a wavelength of 1.54 Å. The single-wavelength anomalous dispersion method was used to successfully solve the structure based on the anomalous signal generated from iodine.
Crystallization and preliminary X-ray analysis of the C-terminal domain of δ-COP, a medium-sized subunit of the COPI complex involved in membrane trafficking
Authors: Deng Kai , Gao Feng , Zheng Peng , Gong Weimin , Sun Zhe
Published: June 2012

Coat protein I (COPI) is a protein complex composed of seven subunits that mediates retrograde transport of proteins and lipids from thecis-Golgi network to the endoplasmic reticulum and intra-Golgi membranes. The medium-sized δ subunit of COPI (δ-COP) is a 57 kDa protein with a C-terminal domain (CTD) and an N-terminal longin domain. Here, the δ-COP CTD was successfully cloned, purified and crystallized. Diffraction data were collected from native and selenomethionyl crystals of δ-COP CTD to resolutions of 2.60 and 2.30 Å, respectively. Both crystals belonged to space groupP21212, with similar unit-cell parameters. The native crystals had unit-cell parametersa= 100.23,b= 136.77,c = 44.39 Å.
Purification, crystallization and preliminary X-ray diffraction analysis of inner membrane complex (IMC) subcompartment protein 1 (ISP1) fromToxoplasma gondii
Authors: Tonkin Michelle L. , Brown Shannon , Beck Josh R. , Bradley Peter J. , Boulanger Martin J.
Published: June 2012

The protozoan parasites of the Apicomplexa phylum are devastating global pathogens. Their success is largely due to phylum-specific proteins found in specialized organelles and cellular structures. The inner membrane complex (IMC) is a unique apicomplexan structure that is essential for motility, invasion and replication. The IMC subcompartment proteins (ISP) have recently been identified inToxoplasma gondiiand shown to be critical for replication, although their specific mechanisms are unknown. Structural characterization ofTgISP1 was pursued in order to identify the fold adopted by the ISPs and to generate detailed insight into how this family of proteins functions during replication. An N-terminally truncated form ofTgISP1 was purified fromEscherichia coli, crystallized and subjected to X-ray diffraction analysis. Two crystal forms ofTgISP1 belonging to space groupsP4132 orP4332 andP212121diffracted to 2.05 and 2.1 Å resolution, respectively.
Capacitor Voltage Regulation in Single-DC-Source Cascaded H-Bridge Multilevel Converters Using Phase-Shift Modulation
Authors: Sepahvand Hossein , Liao Jingsheng , Ferdowsi Mehdi , Corzine Keith
Published: June 2012

Cascaded H-bridge multilevel power electronic converters generally require several dc sources. An alternative option is to replace all the separate dc sources feeding the H-bridge cells with capacitors, leaving only one H-bridge cell with a real dc voltage source. This will yield a cost-effective converter. However, the required capacitor voltage balancing is challenging. In this paper, using the phase-shift modulation approach, a new control method for cascaded H-bridge multilevel converters fed with only one independent dc source is presented. The proposed method has a wide voltage regulation range for the replacement capacitors in the H-bridge cells. Experimental and simulation results support the proposed control method.
Clinical follow‐up study of adult‐onset Still’s disease
Authors: NAGAI Yayoi , HASEGAWA Michiko , OKADA Etsuko , HATTORI Tomoyasu , TAGO Osamu , ISHIKAWA Osamu
Published: June 2012

Eighteen patients with adult‐onset Still’s disease have been followed up for 3–22 years in our department. Initial manifestations were fever with skin rash in 14 patients, fever, skin rash and sore throat in two, skin rash in one and arthralgia in one. During the follow‐up period, typical skin rash was seen in all patients, of them five patients (29%) revealed atypical skin rash simultaneously. Atypical rash included persistent erythema with pigmentation in two, persistent plaques and papules with linear erythema in two and edema of the eyelids mimicking dermatomyositis in one. Persistent papules and plaques revealed histologically characteristic features, such as dyskeratotic keratinocyte and liquefaction degeneration as well as a sparse superficial dermal infiltrate containing scattered neutrophils. In patients of chronic articular type and polycyclic systemic type, atypical skin rash, lymphadenopathy and hyperferritinemia were noted to be significantly higher than those of monocyclic type. These factors might be prognostic factors of adult‐onset Still’s disease in our study.
P19 embryonal carcinoma cells exhibit high sensitivity to botulinum type C and D/C mosaic neurotoxins
Authors: Tsukamoto Kentaro , Arimitsu Hideyuki , Ochi Sadayuki , Nakamura Keiji , Tanaka Yoshikazu , Nuemket Nipawan , Taniguchi Koki , Kozaki Shunji , Tsuji Takao
Published: June 2012

Botulinum neurotoxins (BoNTs) inhibit neurotransmitter release at peripheral nerve terminals. They are serologically classified from A to G, C/D and D/C mosaic neurotoxins forming further subtypes of serotypes C and D. Cultured primary neurons, as well as neuronal cell lines such as PC12 and Neuro‐2a, are often utilized in cell‐based experiments on the toxic action of botulinum toxins. However, there are very few reports of the use of neural cell lines for studying BoNTs/C and D. In addition, the differentiated P19 neuronal cell line, which possesses cholinergic properties, has yet to be tested for its susceptibility to BoNTs. Here, the responsiveness of differentiated P19 cells to BoNT/C and BoNT/DC is reported. Both BoNT/C and BoNT/DC were shown to effectively bind to, and be internalized by, neurons derived from P19 cells. Subsequently, the intracellular substrates for BoNT/C and BoNT/DC were cleaved by treatment of the cells with the toxins in a ganglioside‐dependent manner. Moreover, P19 neurons exhibited high sensitivity to BoNT/C and BoNT/DC, to the same extent as cultured primary neurons. These findings suggest that differentiated P19 cells possess full sensitivity to BoNT/C and BoNT/DC, thus making them a novel susceptible cell line for research into BoNTs.
Tax is a potential molecular target for immunotherapy of adult T‐cell leukemia/lymphoma
Authors: Suzuki Susumu , Masaki Ayako , Ishida Takashi , Ito Asahi , Mori Fumiko , Sato Fumihiko , Narita Tomoko , Ri Masaki , Kusumoto Shigeru , Komatsu Hirokazu , Fukumori Yasuo , Nishikawa Hiroyoshi , Tanaka Yuetsu , Niimi Akio , Inagaki Hiroshi , Iida Shinsuke , Ueda Ryuzo
Published: June 2012

We expanded CTL specific for Tax (a human T‐lymphotropic virus type‐1‐encoded gene product) in vitro from PBMC of several adult T‐cell leukemia/lymphoma (ATL) patients, and document its potential significance as a target for ATL immunotherapy. Tax‐specific CTL responses against tumor cells were restricted by Tax‐expression and the appropriate human leukocyte antigen (HLA) type. Tax‐specific CTL recognized HLA/Tax‐peptide complexes on autologous ATL cells, even when their Tax expression was so low that it could only be detected by RT‐PCR but not by flow cytometry. Recognition resulted in interferon gamma (IFN‐γ) production and target cell lysis. This would be the first report that Tax‐specific CTL from ATL patients specifically recognized and killed autologous tumor cells that expressed Tax. The Tax‐specific CTL responded to as little as 0.01 pM of the corresponding peptide, indicating that their T‐cell receptor avidity was much higher than that of any other CTL recognizing viral or other tumor antigens. This is presumably the reason why the Tax‐specific CTL recognized and killed autologous ATL cells despite their very low Tax expression. In addition, cell cycle analyses and experiments with primary ATL cell‐bearing mice demonstrated that ATL cells present at the site of active cell proliferation, such as in the tumor masses, expressed substantial amounts of Tax, but it was minimally expressed by the tumor cells in a quiescent state, such as in the blood. The present study not only provides a strong rationale for exploiting Tax as a possible target for ATL immunotherapy but also contributes to our understanding of the immunopathogenesis of ATL.
Techniques to increase lymph node harvest from gastrointestinal cancer specimens: a systematic review and meta‐analysis
Authors: Abbassi‐Ghadi Nima , Boshier Piers R , Goldin Robert , Hanna George B
Published: June 2012

Abbassi‐Ghadi N, Boshier P R, Goldin R & Hanna G B (2012) Histopathology61, 531–542  Techniques to increase lymph node harvest from gastrointestinal cancer specimens: a systematic review and meta‐analysis

Aim:   This review aims to compare different histopathological techniques for lymph node harvest from ex‐vivo gastrointestinal cancer specimens and to examine their influence on: (i) lymph node yield; (ii) positive lymph node detection; and (iii) cancer staging.

Method and results:   Systematic review of the English language literature to 10 October 2011, comparing manual nodal dissection to other techniques for lymph node harvest. The methodological quality of included studies was assessed. Twenty‐seven studies, examining fat clearing, methylene blue staining, fat stretching and use of a dedicated pathology assistant, were assessed. The methodological quality of the majority of included studies was poor. Meta‐analysis showed that fat clearing and methylene blue staining increased mean lymph node yield by 13 and 15 nodes, respectively, when compared to manual dissection. Of the 15 studies reporting positive lymph node count, two demonstrated a significant improvement for techniques other than manual dissection. Compared to manual dissection, other techniques were not shown to influence cancer staging.

Conclusion:   This review has shown that fat clearing and methylene blue staining increases the mean lymph node yield from gastrointestinal cancer specimens. There is insufficient evidence to suggest that these techniques increase positive lymph node count or lead to upstaging.
Adaptive landscape genetics: pitfalls and benefits
Authors: PARISOD CHRISTIAN , HOLDEREGGER ROLF
Published: June 2012

Landscape genetics offers a promising framework for assessing the interactions between the environment and adaptive genetic variation in natural populations. A recent workshop held at the University of Neuchatel brought together leading experts in this field to address current insights and future research directions in adaptive landscape genetics. Considerable amounts of genetic and/or environmental data can now be collected, but the forthcoming challenge is to do more with such manna. This requires a markedly better understanding of the genetic variation that is adaptive and prompts for advances in information management together with the development of a balance between theory and data. Moreover, showing the links between landscapes and adaptive genetic variation will ultimately move the field beyond association studies.
Platelet‐activating factor and distinct chemokines are elevated in mucosal biopsies of erosive compared with non‐erosive reflux disease patients and controls
Authors: Altomare A. , Ma J. , Guarino M. P. L. , Cheng L. , Rieder F. , Ribolsi M. , Fiocchi C. , Biancani P. , Harnett K. , Cicala M.
Published: June 2012

Background  A distinction between symptomatic non‐erosive reflux disease (NERD) and erosive esophagitis (EE) patients is supported by the presence of inflammatory response in the mucosa of EE patients, leading to a damage of mucosal integrity. To explore the underlying mechanism of this difference, we assessed inflammatory mediators in mucosal biopsies from EE and NERD patients and compared them with controls.

Methods  Nineteen NERD patients, 15 EE patients, and 16 healthy subjects underwent endoscopy after a 3‐week washout from PPI or H2 antagonists. Biopsies obtained from the distal esophagus were examined by quantitative real‐time polymerase chain reaction (qPCR) and multiplex enzyme‐linked immunosorbent assay for selected chemokines and lyso‐PAF acetyltransferase (LysoPAF‐AT), the enzyme responsible for production of platelet‐activating factor (PAF).

Key Results  Expression of LysoPAF‐AT and multiple chemokines was significantly increased in mucosal biopsies derived from EE patients, when compared with NERD patients and healthy controls. Upregulated chemokines included interleukin 8, eotaxin‐1, ‐2, and ‐3, macrophage inflammatory protein‐1α (MIP‐1α), and monocyte chemoattractant protein‐1 (MCP‐1). LysoPAF‐AT and the chemokine profile in NERD patients were comparable with healthy controls.

Conclusions & Inferences  Levels of selected cytokines and Lyso‐PAF AT were significantly higher in the esophageal mucosa of EE patients compared with NERD and control patients. This difference may explain the distinct inflammatory response occurring in EE patients’ mucosa. In contrast, as no significant differences existed between the levels of all mediators in NERD and control subjects, an inflammatory response does not appear to play a major role in the pathogenesis of the abnormalities found in NERD patients.
The Role of Care‐seeking Delays in Intrauterine Fetal Deaths among ‘Near‐miss’ Women in Herat, Afghanistan
Authors: Hirose Atsumi , Borchert Matthias , Niksear Homa , Alkozai Ahmad S. , Gardiner Julian , Filippi Véronique
Published: June 2012

Background:

Many pregnant women in resource‐poor countries seek care only after developing severe complications during childbirth at home and often reach health facilities in moribund conditions. The objectives were to (i) investigate the association between care‐seeking duration and fetal survival at admission; and (ii) assess the significance of care‐seeking duration in relation to other determinants.

Methods:

Data were analysed for 266 women who were pregnant with a singleton and admitted in life‐threatening conditions to the maternity ward of Herat Regional Hospital in Afghanistan from February 2007 to January 2008. Information about the women's care‐seeking durations, social and financial resources, reproductive factors, household economic status and household types were collected during interviews with the women and their husbands. Information about fetal heartbeats at admission was extracted from the women's medical records.

Results:

Fifty‐four per cent of the women had a decision delay lasting 3 h or more; 69% had a transport delay lasting 3 h or more. Multivariable logistic regression analyses suggest that a decision delay lasting an hour or more increased the odds of fetal death by 6.6 (95% confidence interval [CI] 1.6, 26.3) compared with a delay less than 1 h. A woman's lack of financial autonomy and a distance from her natal home increased the odds of fetal death by 3.1 [95% CI 1.1, 8.4] and 2.5 [95% CI 1.0, 6.3] respectively.

Conclusion:

An integrated approach to improving fetal and maternal health from pre‐pregnancy through childbirth (including increasing women's social and financial resources) is crucial particularly where senior family members act as gatekeepers to women's access to health care.
C3 plants enhance rates of photosynthesis by reassimilating photorespired and respired CO2
Authors: BUSCH FLORIAN A. , SAGE TAMMY L. , COUSINS ASAPH B. , SAGE ROWAN F.
Published: June 2012

Photosynthetic carbon gain in plants using the C3 photosynthetic pathway is substantially inhibited by photorespiration in warm environments, particularly in atmospheres with low CO2 concentrations. Unlike C4 plants, C3 plants are thought to lack any mechanism to compensate for the loss of photosynthetic productivity caused by photorespiration. Here, for the first time, we demonstrate that the C3 plants rice and wheat employ a specific mechanism to trap and reassimilate photorespired CO2. A continuous layer of chloroplasts covering the portion of the mesophyll cell periphery that is exposed to the intercellular air space creates a diffusion barrier for CO2 exiting the cell. This facilitates the capture and reassimilation of photorespired CO2 in the chloroplast stroma. In both species, 24–38% of photorespired and respired CO2 were reassimilated within the cell, thereby boosting photosynthesis by 8–11% at ambient atmospheric CO2 concentration and 17–33% at a CO2 concentration of 200 µmol mol−1. Widespread use of this mechanism in tropical and subtropical C3 plants could explain why the diversity of the world's C3 flora, and dominance of terrestrial net primary productivity, was maintained during the Pleistocene, when atmospheric CO2 concentrations fell below 200 µmol mol−1.
Invasive Ants Alter Foraging and Parental Behaviors of a Native Bird
Authors: Ligon Russell A. , Siefferman Lynn , Hill Geoffrey E.
Published: June 2012

Introduced species can exert outsized impacts on native biota through both direct (predation) and indirect (competition) effects. Ants frequently become established in new areas after being transported by humans across traditional biological or geographical barriers, and a prime example of such establishment is the red imported fire ant (Solenopsis invicta). Introduced to North America in the 1930s, red imported fire ants are now firmly established throughout the southeastern United States. Although these invasive predators can dramatically impact native arthropods, their effect on vertebrates through resource competition is essentially unknown. Using a paired experimental design, we compared patterns of foraging and rates of provisioning for breeding eastern bluebirds (Sialia sialis) in unmanipulated (control) territories to those in adjacent (treated) territories where fire ants were experimentally reduced. Bluebirds inhabiting treated territories foraged nearer their nests and provisioned offspring more frequently than bluebirds inhabiting control territories with unmanipulated fire ant levels. Additionally, nestlings from treated territories were in better condition than those from control territories, though these differences were largely confined to early development. The elimination of significant differences in body condition toward the end of the nestling period suggests that bluebird parents in control territories were able to make up the food deficit caused by fire ants, potentially by working harder to adequately provision their offspring. The relationship between fire ant abundance and bluebird behavior hints at the complexity of ecological communities and suggests negative effects of invasive species are not limited to taxa with which they have direct contact.
Screening for resistance to black rot in Brassica oleracea crops
Authors: Lema Margarita , Velasco Pablo , Soengas Pilar , Francisco Marta , Cartea María Elena
Published: June 2012

With 4 tables

Xanthomonas campestris pv. campestris (Xcc), causal agent of black rot, is widely distributed around the world in cabbage and kale crops causing severe yield losses. Nine races of the pathogen were described, being races 1 and 4 the most aggressive and widespread. The objective of this work was to identify new sources of resistance to races 1 and 4 of Xcc in different Brassica oleracea crops. Two hundred and twenty‐one landraces and 35 commercial and improved cultivars were evaluated. Most accessions were susceptible to both races, except cabbage cultivars ‘Balón’ and ‘Quintal de Alsacia’ and kale landrace MBG‐BRS0070, which showed some plants with different level of resistance to races 1 and 4, thus indicating that race‐non‐specific resistance can be involved. Kale landrace MBG‐BRS0286 showed an intermediate mean disease score for race 1. These accessions can be crossed with cabbage cultivars and may provide new combinations of resistance genes with protection against black rot.
Postpartum Variations of Plasma IGF and IGFBPs, Oocyte Production and Quality in Dairy Cows: Relationships With Parity and Subsequent Fertility
Authors: Grimard B , Marquant‐Leguienne B , Remy D , Richard C , Nuttinck F , Humblot P , Ponter AA
Published: June 2012

Contents

The aim of this study was to determine whether postpartum variations of plasma IGF‐1 and IGFBP concentrations, oocyte production and quality were related to parity and subsequent conception rate in Holstein dairy cows. Holstein dairy cows [10 primiparous (PP) and 22 multiparous (MP)] were allotted in six batches and sampled once weekly between calving and oestrous synchronization treatment started at 71.2 ± 2.0 days postpartum. During the 3 weeks before treatment, ovum pick‐up (OPU) was performed twice weekly. Oocytes were scored on a 4‐point scale, and oocytes from OPU1, 3 and 5 were fertilized in vitro. Seventeen cows became pregnant after first and second AI and were considered as fertile (F), while the others were considered to be subfertile (SF). Logistic regression was carried out to investigate the relationships between repeated measurements and fertility including parity and batch effects in the models. Likelihood of fertility significantly increased when plasma urea and IGFBP‐3 concentrations decreased and was higher in PP compared with MP cows. There was a trend for fertility to increase when plasma IGF‐1 concentrations increased (p = 0.07). In vitro cleavage and development rates were similar between SF and F cows (46.4% and 28.3% in SF vs 55.0% and 22.1% in F). Parity had an effect on plasma IGF‐1 concentrations (PP: 61.65 ± 2.67 vs MP: 41.63 ± 5.81 ng/ml, p < 0.001), mean number of follicles aspirated per session (PP: 5.7 ± 1.3 vs MP: 9.5 ± 0.8, p < 0.05) and fertility (PP: 8/10 = 80% vs MP: 9/22 = 41%, p < 0.05) but not on the number of oocytes recovered per session nor their quality. In conclusion, postpartum plasma urea and IGFBP‐3 concentrations, but not oocyte production and quality before breeding, were related to subsequent conception rate in our experimental design. Parity had a significant effect on energy status, follicular growth and fertility and needs to be considered when investigating relationships between nutrition and reproduction.