bluuebunny/crossref_metadata_2025 huggingface dataset:
Of Tournaments and Temptations: An Analysis of Managerial Incentives in the Mutual Fund Industry
Authors: BROWN KEITH C. , HARLOW W. V. , STARKS LAURA T.
Published: April 2012

We test the hypothesis that when their compensation is linked to relative performance, managers of investment portfolios likely to end up as “losers” will manipulate fund risk differently than those managing portfolios likely to be “winners.” An empirical investigation of the performance of 334 growth‐oriented mutual funds during 1976 to 1991 demonstrates that mid‐year losers tend to increase fund volatility in the latter part of an annual assessment period to a greater extent than mid‐year winners. Furthermore, we show that this effect became stronger as industry growth and investor awareness of fund performance increased over time.
Real Interest Rates and Inflation: An Ex‐Ante Empirical Analysis
Authors: KANDEL SHMUEL , OFER AHARON R. , SARIG ODED
Published: April 2012

We develop a method of measuring ex‐ante real interest rates using prices of index and nominal bonds. Employing this method and newly available data, we directly test the Fisher hypothesis that the real rate of interest is independent of inflation expectations. We find a negative correlation between ex‐ante real interest rates and expected inflation. This contradicts the Fisher hypothesis but is consistent with the theories of Mundell and Tobin, Darby and Feldstein, and Stulz. We also find that nominal interest rates include an inflation risk premium that is positively related to a proxy for inflation uncertainty.
Novel meningococcal 4CMenB vaccine antigens – prevalence and polymorphisms of the encoding genes in Neisseria gonorrhoeae
Authors: Hadad Ronza , Jacobsson Susanne , Pizza Mariagrazia , Rappuoli Rino , Fredlund Hans , Olcén Per , Unemo Magnus
Published: April 2012

The first cross‐protective Neisseria meningitidis vaccine (focus on serogroup B), the protein‐based 4 component meningococcus serogroup B (4CMenB), includes the New Zealand outer membrane vesicle and three main genome‐derived neisserial antigens (GNAs). These GNAs are fHbp (fused to GNA2091), NHBA (fused to GNA1030) and NadA. In this study, the prevalence and polymorphisms of the nucleotide and amino acid sequences of the 4CMenB antigens in a temporally and geographically diverse collection of N. gonorrhoeae isolates (n = 111) were investigated. All the examined GNA genes, except the nadA gene, were present in all gonococcal isolates. However, 25 isolates contained premature stop codons in the fHbp gene and/or the nhba gene, resulting in truncated proteins. Compared with the 4CMenB antigen sequences in reference strain MC58, the gonococcal strains displayed 67.0–95.4% and 60.9–94.9% identity in nucleotide sequence and amino acid sequence, respectively, in the equivalent GNA antigens. The absence of NadA, lack of universal expression of fHbp and NHBA and the uncertainty regarding the surface exposure of fHbp as well as the function of NHBA in N. gonorrhoeae will likely limit the use of the identical 4CMenB antigens in a gonococcal vaccine. However, possible cross‐immunity of 4CMenB with gonococci and expression and function of the equivalent gonococcal GNAs, as well as of more appropriate GNAs for a gonococcal vaccine, need to be further examined.
Antibiotic susceptibility in prostate‐derived Propionibacterium acnes isolates
Authors: Olsson Jan , Davidsson Sabina , Unemo Magnus , Mölling Paula , Andersson Swen‐Olov , Andrén Ove , Söderquist Bo , Sellin Mats , Elgh Fredrik
Published: April 2012

The aim of this study was to determine antibiotic susceptibility of Propionibacterium acnes isolates from prostate. Prostate‐derived P. acnes isolates (n = 24, Umeå & Örebro, Sweden, 2007–2010) and a panel of control strains (n = 25, Sweden) collected from skin and deep infections were assessed for resistance to penicillin G, piperacillin–tazobactam, imipenem, gentamicin, azithromycin, erythromycin, vancomycin, ciprofloxacin, moxifloxacin, tetracycline, tigecycline, fusidic acid, clindamycin, rifampicin, linezolid, daptomycin, trimethoprim–sulfamethoxazole, and metronidazole. In addition, the isolates were tested for inducible clindamycin resistance. All prostate derived P. acnes isolates displayed wild‐type distribution of MIC‐values, without evidence of acquired resistance. In the reference panel, 5 of 25 isolates had acquired macrolide resistance with cross‐resistance to azithromycin, clindamycin, and erythromycin. In addition, one of these isolates was resistant to tetracycline.
Nelfinavir inhibits regulated intramembrane proteolysis of sterol regulatory element binding protein‐1 and activating transcription factor 6 in castration‐resistant prostate cancer
Authors: Guan Min , Fousek Kristen , Chow Warren A.
Published: April 2012

Nelfinavir induces apoptosis in liposarcoma by inhibiting site‐2 protease (S2P) activity, which leads to suppression of regulated intramembrane proteolysis. We postulate similar effects in castration‐resistant prostate cancer because it exhibits a lipogenic phenotype. Nelfinavir inhibited androgen receptor activation in androgen‐sensitive prostate cancer and the nuclear translocation of the fusion proteins sterol regulatory element binding protein‐1 (SREBP‐1)–enhanced green fluorescence protein (EGFP) and activating transcription factor 6 (ATF6)–EGFP in castration‐resistant prostate cancer cells, viewed under confocal microscopy. Nelfinavir and site‐1 protease (S1P) and S2P small interfering RNAs (siRNAs) reduced the proliferation of castration‐resistant prostate cancer and induced apoptosis, which was opposed by autophagy. Inhibition of autophagy with hydroxychloroquine was additive to the apoptotic effect of nelfinavir. Western blotting of S1P and S2P siRNA knockdown and/or nelfinavir‐treated cells confirmed the accumulation of precursor SREBP‐1 and ATF6. 3,4‐Dichloroisocoumarin, an S1P inhibitor, did not affect SREBP‐1 processing. In contrast, 1,10‐phenanthroline, an S2P inhibitor, reproduced the nelfinavir‐treated molecular and biological phenotype. Nelfinavir‐mediated inhibition of regulated intramembrane proteolysis led to the accumulation of unprocessed SREBP‐1 and ATF6. This resulted in sequential endoplasmic reticulum stress, inhibition of the unfolded protein response, reduced fatty acid synthase expression and apoptosis, which was countered by autophagy. Inhibition of autophagy was at least additive to this pro‐apoptotic effect. These findings provide new insights into nelfinavir‐induced endoplasmic reticulum stress and cancer cell death, and lead us to propose investigating its clinical activity in castration‐resistant prostate cancer. This report validates S2P as a therapeutic target in castration‐resistant prostate cancer.
Phenological growth stages of the golden penda tree (Xanthostemon chrysanthus)
Authors: Ahmad Nazarudin M.R. , Tsan F.Y. , Normaniza O. , Adzmi Y.
Published: April 2012

Biologishe Bundesanstalt, Bundessortenamt and CHemical Industry (BBCH) scales are a system for the uniform coding of phenologically similar growth stages of plants, using a two‐digit decimal code. This paper describes the phenological growth stages of a landscape tree, Xanthostemon chrysanthus using the BBCH‐scale. Detailed descriptions of principal growth stages for bud, leaf and shoot development, inflorescence emergence, flowering, fruit development, maturity or ripening of fruit and seed, and other secondary growth stages are provided. The developed scale will be of great help to landscape managers and researchers for better communication, more efficient planning of management practices and experiments of flowering trees.
Pollinating fig wasp Ceratosolen solmsi adjusts the offspring sex ratio to other foundresses
Authors: Hu Hao‐Yuan , Chen Zhong‐Zheng , Jiang Zi‐Feng , Huang Da‐Wei , Niu Li‐Ming , Fu Yue‐Guan
Published: April 2012

Abstract  Local mate competition theory predicts that offspring sex ratio in pollinating fig wasps is female‐biased when there is only one foundress, and increased foundress density results in increased offspring sex ratio. Information of other foundresses and clutch size have been suggested to be the main proximate explanations for sex ratio adjustment under local mate competition. Our focus was to show the mechanism of sex ratio adjustment in a pollinating fig wasp, Ceratosolen solmsi Mayr, an obligate pollinator of the functionally dioecious fig, Ficus hispida Linn., with controlled experiments in the field. First, we obtained offspring from one pollinator and offspring at different oviposition sequences, and found that offspring sex ratio decreased with clutch size, and pollinators produced most of their male offspring at the start of bouts, followed by mostly females. Second, we found that offspring sex ratio increased with foundress density, and pollinators did adjust their offspring sex ratio to other females in the oviposition patches. We suggest that when oviposition sites are not limited, pollinators will mainly adjust their offspring sex ratio to other foundresses independent of clutch size changes, whereas adjusting clutch size may be used to adjust sex ratio when oviposition sites are limited.
A multicenter study of clinical and laboratory findings of palindromic rheumatism in Iran
Authors: Khabbazi Alireza , Hajialiloo Mehrzad , Kolahi Sousan , Soroosh Mohsen , Esalatmanesh Kamal , Sharif Sakinehkhatoon
Published: April 2012

Objective

Considering demographic, clinical and laboratory characteristics of palindromic rheumatism and finding out the difference between rheumatoid factor and anti‐cyclic citrullinated peptide positive and negative cases of palindromic rheumatism in Iran.

Methods

In this cross‐sectional and multicenter study, patients with intermittent arthritis were evaluated by a rheumatologist for palindromic rheumatism. Diagnosis of palindromic rheumatism was made after ruling out other causes of recurrent arthritis, and the fulfillment of the Pasero and Barbieri criteria. Then the demographic and clinical characteristics were recorded and compared in rheumatoid factor and anti‐cyclic citrullinated peptide positive and negative patients.

Results

Of the 69 patients with palindromic rheumatism, 35 were men and 34 were women. The mean ages of the patients at the time of diagnosis were 38.5 ± 14.6 years. The mean attack intervals and duration of the attacks were 66.3 ± 38.4 days and 3.9 ± 3.3 days, respectively. The most commonly involved joints were knees, metacarpophalangeals and proximal interphalangeals. Rheumatoid factor in 46.4% and anti‐cyclic citrullinated peptide in 42% of the cases were positive. Palindromic patients with positive anti‐cyclic citrullinated peptide had higher age, shorter duration of disease, shorter duration of attacks, more frequent attacks and more metacarpophalangeal joints involvement.

Conclusions

Rheumatoid factor and anti‐cyclic citrullinated peptides were found in a high proportion of patients with palindromic rheumatism. The clinical features of the disease in our study were different in rheumatoid factor and anti‐cyclic citrullinated peptide positive and negative patients.
Novel Metagenome-Derived, Cold-Adapted Alkaline Phospholipase with Superior Lipase Activity as an Intermediate between Phospholipase and Lipase
Authors: Lee Mi-Hwa , Oh Ki-Hoon , Kang Chul-Hyung , Kim Ji-Hoon , Oh Tae-Kwang , Ryu Choong-Min , Yoon Jung-Hoon
Published: April 2012

A novel lipolytic enzyme was isolated from a metagenomic library obtained from tidal flat sediments on the Korean west coast. Its putative functional domain, designated MPlaG, showed the highest similarity to phospholipase A from
Grimontia hollisae
CIP 101886, though it was screened from an emulsified tricaprylin plate. Phylogenetic analysis showed that MPlaG is far from family I.6 lipases, including
Staphylococcus hyicus
lipase, a unique lipase which can hydrolyze phospholipids, and is more evolutionarily related to the bacterial phospholipase A
1
family. The specific activities of MPlaG against olive oil and phosphatidylcholine were determined to be 2,957 ± 144 and 1,735 ± 147 U mg
−1
, respectively, which means that MPlaG is a lipid-preferred phospholipase. Among different synthetic esters, triglycerides, and phosphatidylcholine, purified MPlaG exhibited the highest activity toward
p
-nitrophenyl palmitate (C
16
), tributyrin (C
4
), and 1,2-dihexanoyl-phosphatidylcholine (C
8
). Finally, MPlaG was identified as a phospholipase A
1
with lipase activity by cleavage of the
sn
-1 position of OPPC, interfacial activity, and triolein hydrolysis. These findings suggest that MPlaG is the first experimentally characterized phospholipase A
1
with lipase activity obtained from a metagenomic library. Our study provides an opportunity to improve our insight into the evolution of lipases and phospholipases.
Involvement of Clostridium botulinum ATCC 3502 Sigma Factor K in Early-Stage Sporulation
Authors: Kirk David G. , Dahlsten Elias , Zhang Zhen , Korkeala Hannu , Lindström Miia
Published: April 2012

A key survival mechanism of
Clostridium botulinum
, the notorious neurotoxic food pathogen, is the ability to form heat-resistant spores. While the genetic mechanisms of sporulation are well understood in the model organism
Bacillus subtilis
, nothing is known about these mechanisms in
C. botulinum.
Using the ClosTron gene-knockout tool,
sigK
, encoding late-stage (stage IV) sporulation sigma factor K in
B. subtilis
, was disrupted in
C. botulinum
ATCC 3502 to produce two different mutants with distinct insertion sites and orientations. Both mutants were unable to form spores, and their elongated cell morphology suggested that the sporulation pathway was blocked at an early stage. In contrast,
sigK
-complemented mutants sporulated successfully. Quantitative real-time PCR analysis of
sigK
in the parent strain revealed expression at the late log growth phase in the parent strain. Analysis of
spo0A
, encoding the sporulation master switch, in the
sigK
mutant and the parent showed significantly reduced relative levels of
spo0A
expression in the
sigK
mutant compared to the parent strain. Similarly,
sigF
showed significantly lower relative transcription levels in the
sigK
mutant than the parent strain, suggesting that the sporulation pathway was blocked in the
sigK
mutant at an early stage. We conclude that σ
K
is essential for early-stage sporulation in
C. botulinum
ATCC 3502, rather than being involved in late-stage sporulation, as reported for the sporulation model organism
B. subtilis
. Understanding the sporulation mechanism of
C. botulinum
provides keys to control the public health risks that the spores of this dangerous pathogen cause through foods.
Genetic and Functional Characterization of Cyclic Lipopeptide White-Line-Inducing Principle (WLIP) Production by Rice Rhizosphere Isolate Pseudomonas putida RW10S2
Authors: Rokni-Zadeh Hassan , Li Wen , Sanchez-Rodriguez Aminael , Sinnaeve Davy , Rozenski Jef , Martins José C. , De Mot René
Published: April 2012

The secondary metabolite mediating the GacS-dependent growth-inhibitory effect exerted by the rice rhizosphere isolate
Pseudomonas putida
RW10S2 on phytopathogenic
Xanthomonas
species was identified as white-line-inducing principle (WLIP), a member of the viscosin group of cyclic lipononadepsipeptides. WLIP producers are commonly referred to by the taxonomically invalid name “
Pseudomonas reactans
,” based on their capacity to reveal the presence of a nearby colony of
Pseudomonas tolaasii
by inducing the formation of a visible precipitate (“white line”) in agar medium between both colonies. This phenomenon is attributed to the interaction of WLIP with a cyclic lipopeptide of a distinct structural group, the fungitoxic tolaasin, and has found application as a diagnostic tool to identify tolaasin-producing bacteria pathogenic to mushrooms. The genes encoding the WLIP nonribosomal peptide synthetases WlpA, WlpB, and WlpC were identified in two separate genomic clusters (
wlpR-wlpA
and
wlpBC
) with an operon organization similar to that of the viscosin, massetolide, and entolysin biosynthetic systems. Expression of
wlpR
is dependent on
gacS
, and the encoded regulator of the LuxR family (WlpR) activates transcription of the biosynthetic genes and the linked export genes, which is not controlled by the RW10S2 quorum-sensing system PmrR/PmrI. In addition to linking the known phenotypes of white line production and hemolytic activity of a WLIP producer with WLIP biosynthesis, additional properties of ecological relevance conferred by WLIP production were identified, namely, antagonism against
Xanthomonas
and involvement in swarming and biofilm formation.
No Beneficial Effects Evident for Enterococcus faecium NCIMB 10415 in Weaned Pigs Infected with Salmonella enterica Serovar Typhimurium DT104
Authors: Kreuzer Susanne , Janczyk Pawel , Aßmus Jens , Schmidt Michael F. G. , Brockmann Gudrun A. , Nöckler Karsten
Published: April 2012

Salmonella enterica
serovar Typhimurium DT 104 is the major pathogen for salmonellosis outbreaks in Europe. We tested if the probiotic bacterium
Enterococcus faecium
NCIMB 10415 can prevent or alleviate salmonellosis. Therefore, piglets of the German Landrace breed that were treated with
E. faecium
(
n
= 16) as a feed additive and untreated controls (
n
= 16) were challenged with
S.
Typhimurium 10 days after weaning. The presence of salmonellae in feces and selected organs, as well as the immune response, were investigated. Piglets treated with
E. faecium
gained less weight than control piglets (
P
= 0.05). The feeding of
E. faecium
had no effect on the fecal shedding of salmonellae and resulted in a higher abundance of the pathogen in tonsils of all challenged animals. The specific (anti-
Salmonella
IgG) and nonspecific (haptoglobin) humoral immune responses as well as the cellular immune response (T helper cells, cytotoxic T cells, regulatory T cells, γδ T cells, and B cells) in the lymph nodes, Peyer's patches of different segments of the intestine (jejunal and ileocecal), the ileal papilla, and in the blood were affected in the course of time after infection (
P
< 0.05) but not by the
E. faecium
treatment. These results led to the conclusion that
E. faecium
may not have beneficial effects on the performance of weaned piglets in the case of
S.
Typhimurium infection. Therefore, we suggest a critical discussion and reconsideration of
E. faecium
NCIMB 10415 administration as a probiotic for pigs.
Temperature-Dependent Survival of Hepatitis A Virus during Storage of Contaminated Onions
Authors: Sun Y. , Laird D. T. , Shieh Y. C.
Published: April 2012

Pre- or postharvest contamination of green onions by hepatitis A virus (HAV) has been linked to large numbers of food-borne illnesses. Understanding HAV survival in onions would assist in projecting the risk of the disease associated with their consumption. This study defined HAV inactivation rates in contaminated green onions contained in air-permeable, moisture-retaining high-density polyethylene packages that were stored at 3, 10, 14, 20, 21, 22, and 23°C. A protocol was established to recover HAV from whole green onions, with 31% as the average recovery by infectivity assay. Viruses in eluates were primarily analyzed by a 6-well plaque assay on FRhK-4 cells. Eight storage trials, including two trials at 3°C, were conducted, with 3 to 7 onion samples per sampling and 4 to 7 samplings per trial. Linear regression correlation (
r
2
= 0.80 to 0.98) was observed between HAV survival and storage time for each of the 8 trials, held at specific temperatures. Increases in the storage temperature resulted in greater HAV inactivation rates, e.g., a reduction of 0.033 log PFU/day at 3.4 ± 0.3°C versus 0.185 log PFU/day at 23.4 ± 0.7°C. Thus, decimal reduction time (
D
) values of 30, 14, 11, and 5 days, respectively, were obtained for HAV in onions stored at 3, 10, 14, and 23°C. Further regression analysis determined that 1 degree Celsius increase would increase inactivation of HAV by 0.007 log PFU/day in onions (
r
2
= 0.97). The data suggest that natural degradation of HAV in contaminated fresh produce is minimal and that a preventive strategy is critical to produce safety. The results are useful in predicting the risks associated with HAV contamination in fresh produce.
Histopathological Effects of the Yen-Tc Toxin Complex from Yersinia entomophaga MH96 (Enterobacteriaceae) on the Costelytra zealandica (Coleoptera: Scarabaeidae) Larval Midgut
Authors: Marshall Sean D. G. , Hares Michelle C. , Jones Sandra A. , Harper Lincoln A. , Vernon James R. , Harland Duane P. , Jackson Trevor A. , Hurst Mark R. H.
Published: April 2012

Yersinia entomophaga
MH96, which was originally isolated from the New Zealand grass grub,
Costelytra zealandica
, produces an orally active proteinaceous toxin complex (Yen-Tc), and this toxin is responsible for mortality in a range of insect species, mainly within the Coleoptera and Lepidoptera. The genes encoding Yen-Tc are members of the toxin complex (Tc) family, with orthologs identified in several other bacterial species. As the mechanism of Yen-Tc activity remains unknown, a histopathological examination of
C. zealandica
larvae was undertaken in conjunction with cultured cells to identify the effects of Yen-Tc and to distinguish the contributions that its individual subunit components make upon intoxication. A progressive series of events that led to the deterioration of the midgut epithelium was observed. Additionally, experiments using a cell culture assay system were carried out to determine the cellular effects of intoxication on cells after topical application and the transient expression of Yen-Tc and its individual components. While observations were broadly consistent with those previously reported for other Tc family members, some differences were noted. In particular, the distinct stepwise disintegration of the midgut shared features associated with both apoptosis and necrotic programmed cell death pathways. Second, we observed, for the first time, a contribution of toxicity from two chitinases associated with the Yen-Tc complex. Our findings were suggestive of the activities encoded within the subunit components of Yen-Tc targeting different sites along putative programmed cell death pathways. Given the observed broad host range for Yen-Tc, these targeted loci are likely to be widely shared among insects.
Lactobacillus casei Ferments the
N
-Acetylglucosamine Moiety of Fucosyl-α-1,3-
N
-Acetylglucosamine and Excretes
l
-Fucose
Authors: Rodríguez-Díaz Jesús , Rubio-del-Campo Antonio , Yebra María J.
Published: April 2012

We have previously characterized from
Lactobacillus casei
BL23 three α-
l
-fucosidases, AlfA, AlfB, and AlfC, which hydrolyze
in vitro
natural fucosyl-oligosaccharides. In this work, we have shown that
L. casei
is able to grow in the presence of fucosyl-α-1,3-
N
-acetylglucosamine (Fuc-α-1,3-GlcNAc) as a carbon source. Interestingly,
L. casei
excretes the
l
-fucose moiety during growth on Fuc-α-1,3-GlcNAc, indicating that only the
N
-acetylglucosamine moiety is being metabolized. Analysis of the genomic sequence of
L. casei
BL23 shows that downstream from
alfB
, which encodes the α-
l
-fucosidase AlfB, a gene,
alfR
, that encodes a transcriptional regulator is present. Divergently from
alfB
, three genes,
alfEFG
, that encode proteins with homology to the enzyme IIAB (EIIAB), EIIC, and EIID components of a mannose-class phosphoenolpyruvate:sugar phosphotransferase system (PTS) are present. Inactivation of either
alfB
or
alfF
abolishes the growth of
L. casei
on Fuc-α-1,3-GlcNAc. This proves that AlfB is involved in Fuc-α-1,3-GlcNAc metabolism and that the transporter encoded by
alfEFG
participates in the uptake of this disaccharide. A mutation in the PTS general component enzyme I does not eliminate the utilization of Fuc-α-1,3-GlcNAc, suggesting that the transport via the PTS encoded by
alfEFG
is not coupled to phosphorylation of the disaccharide. Transcriptional analysis with
alfR
and
ccpA
mutants shows that the two gene clusters
alfBR
and
alfEFG
are regulated by substrate-specific induction mediated by the inactivation of the transcriptional repressor AlfR and by carbon catabolite repression mediated by the catabolite control protein A (CcpA). This work reports for the first time the characterization of the physiological role of an α-
l
-fucosidase in lactic acid bacteria and the utilization of Fuc-α-1,3-GlcNAc as a carbon source for bacteria.
Synthesis of Chiral Cyanohydrins by Recombinant Escherichia coli Cells in a Micro-Aqueous Reaction System
Authors: Scholz Kathrin Emmi , Okrob Daniel , Kopka Benita , Grünberger Alexander , Pohl Martina , Jaeger Karl-Erich , Krauss Ulrich
Published: April 2012

Synthesis of chiral cyanohydrins is performed in a monophasic micro-aqueous reaction system using whole recombinant
Escherichia coli
cells expressing the
Arabidopsis thaliana
hydroxynitrile lyase (
At
HNL). Microscopy studies employing a fusion of
At
HNL with a flavin-based fluorescent protein (FbFP) reveal that the cells remain intact in the reaction system.
Evidence for Lack of Acquisition of Tolerance in Salmonella enterica Serovar Typhimurium ATCC 14028 after Exposure to Subinhibitory Amounts of Origanum vulgare L. Essential Oil and Carvacrol
Authors: Luz Isabelle da Silva , Gomes Neto Nelson Justino , Tavares Adassa Gama , Nunes Pollyana Campos , Magnani Marciane , de Souza Evandro Leite
Published: April 2012

Overnight exposure of
Salmonella enterica
serovar Typhimurium to sublethal amounts of
Origanum vulgare
essential oil (OV) and carvacrol (CAR) did not result in direct and cross-bacterial protection. Cells subcultured with increasing amounts of OV or CAR survived up to the MIC of either compound, revealing few significant changes in bacterial susceptibility.
Differential Regulation by Organic Compounds and Heavy Metals of Multiple Laccase Genes in the Aquatic Hyphomycete Clavariopsis aquatica
Authors: Solé Magali , Müller Ines , Pecyna Marek J. , Fetzer Ingo , Harms Hauke , Schlosser Dietmar
Published: April 2012

To advance the understanding of the molecular mechanisms controlling microbial activities involved in carbon cycling and mitigation of environmental pollution in freshwaters, the influence of heavy metals and natural as well as xenobiotic organic compounds on laccase gene expression was quantified using quantitative real-time PCR (qRT-PCR) in an exclusively aquatic fungus (the aquatic hyphomycete
Clavariopsis aquatica
) for the first time. Five putative laccase genes (
lcc1
to
lcc5
) identified in
C. aquatica
were differentially expressed in response to the fungal growth stage and potential laccase inducers, with certain genes being upregulated by, e.g., the lignocellulose breakdown product vanillic acid, the endocrine disruptor technical nonylphenol, manganese, and zinc.
lcc4
is inducible by vanillic acid and most likely encodes an extracellular laccase already excreted during the trophophase of the organism, suggesting a function during fungal substrate colonization. Surprisingly, unlike many laccases of terrestrial fungi, none of the
C. aquatica
laccase genes was found to be upregulated by copper. However, copper strongly increases extracellular laccase activity in
C. aquatica
, possibly due to stabilization of the copper-containing catalytic center of the enzyme. Copper was found to half-saturate laccase activity already at about 1.8 μM, in favor of a fungal adaptation to low copper concentrations of aquatic habitats.
High-Resolution Analysis of Gut Environment and Bacterial Microbiota Reveals Functional Compartmentation of the Gut in Wood-Feeding Higher Termites (Nasutitermes spp.)
Authors: Köhler Tim , Dietrich Carsten , Scheffrahn Rudolf H. , Brune Andreas
Published: April 2012

Higher termites are characterized by a purely prokaryotic gut microbiota and an increased compartmentation of their intestinal tract. In soil-feeding species, each gut compartment has different physicochemical conditions and is colonized by a specific microbial community. Although considerable information has accumulated also for wood-feeding species of the genus
Nasutitermes
, including cellulase activities and metagenomic data, a comprehensive study linking physicochemical gut conditions with the structure of the microbial communities in the different gut compartments is lacking. In this study, we measured high-resolution profiles of H
2
, O
2
, pH, and redox potential in the gut of
Nasutitermes corniger
termites, determined the fermentation products accumulating in the individual gut compartments, and analyzed the bacterial communities in detail by pyrotag sequencing of the V3-V4 region of the 16S rRNA genes. The dilated hindgut paunch (P3 compartment) was the only anoxic gut region, showed the highest density of bacteria, and accumulated H
2
to high partial pressures (up to 12 kPa). Molecular hydrogen is apparently produced by a dense community of
Spirochaetes
and
Fibrobacteres
, which also dominate the gut of other
Nasutitermes
species. All other compartments, such as the alkaline P1 compartment (average pH, 10.0), showed high redox potentials and comprised small but distinct populations characteristic for each gut region. In the crop and the posterior hindgut compartments, the community was even more diverse than in the paunch. Similarities in the communities of the posterior hindgut and crop suggested that proctodeal trophallaxis or coprophagy also occurs in higher termites. The large sampling depths of pyrotag sequencing in combination with the determination of important physicochemical parameters allow cautious conclusions concerning the functions of particular bacterial lineages in the respective gut sections to be drawn.
Elevated Enterotoxin A Expression and Formation in Staphylococcus aureus and Its Association with Prophage Induction
Authors: Cao Rong , Zeaki Nikoleta , Wallin-Carlquist Nina , Skandamis Panagiotis N. , Schelin Jenny , Rådström Peter
Published: April 2012

Staphylococcus aureus
strains producing the bacteriophage-encoded staphylococcal enterotoxin A (SEA) were divided into two groups, high- and low-SEA-producing strains, based on the amount of SEA produced. After growth under favorable conditions in batch cultures, 10 of the 21 strains tested produced more than 1,000 ng/ml SEA, and 9 strains produced less than 10 ng/ml SEA; two enterotoxigenic strains, MRSA252 and Newman, produced intermediate levels of SEA (around 450 ng/ml). The differences in the production of SEA were found to be associated with the expression level of
sea
and whether the strains hosted the
sea
1
or
sea
2
version. Furthermore, differences in nucleotide sequence in the
Siphoviridae
phage region showed two clonal lineages of the high-SEA-producing strains. One of these lines was correlated with the capacity for a massive increase in SEA levels by prophage induction as demonstrated using mitomycin C (MC). This was also confirmed by the occurrence of additional
sea
expression, presumed to be initiated by a latent phage promoter located upstream of the endogenous
sea
promoter. Remarkably, the SEA level was increased up to 10-fold in some strains due to prophage induction. The low-SEA-producing group and the high-SEA-producing subgroup lacking phage-activated
sea
transcription showed no increase in SEA formation after the addition of MC. This study demonstrates that
sea
expression in enterotoxigenic strains is correlated with the clonal lineage of
sea
-carrying phages. The high-SEA-producing group, in particular the prophage-inducible
sea
1
group, may be more relevant to staphylococcal food poisoning than the low-SEA-producing group, harboring mainly
sea
2
.
Implications of Microfauna-Host Interactions for Trypanosome Transmission Dynamics in Glossina fuscipes fuscipes in Uganda
Authors: Alam Uzma , Hyseni Chaz , Symula Rebecca E. , Brelsfoard Corey , Wu Yineng , Kruglov Oleg , Wang Jingwen , Echodu Richard , Alioni Victor , Okedi Loyce M. , Caccone Adalgisa , Aksoy Serap
Published: April 2012

Tsetse flies (Diptera: Glossinidae) are vectors for African trypanosomes (Euglenozoa: kinetoplastida), protozoan parasites that cause African trypanosomiasis in humans (HAT) and nagana in livestock. In addition to trypanosomes, two symbiotic bacteria (
Wigglesworthia glossinidia
and
Sodalis glossinidius
) and two parasitic microbes,
Wolbachia
and a salivary gland hypertrophy virus (SGHV), have been described in tsetse. Here we determined the prevalence of and coinfection dynamics between
Wolbachia
, trypanosomes, and SGHV in
Glossina fuscipes fuscipes
in Uganda over a large geographical scale spanning the range of host genetic and spatial diversity. Using a multivariate analysis approach, we uncovered complex coinfection dynamics between the pathogens and statistically significant associations between host genetic groups and pathogen prevalence. It is important to note that these coinfection dynamics and associations with the host were not apparent by univariate analysis. These associations between host genotype and pathogen are particularly evident for
Wolbachia
and SGHV where host groups are inversely correlated for
Wolbachia
and SGHV prevalence. On the other hand, trypanosome infection prevalence is more complex and covaries with the presence of the other two pathogens, highlighting the importance of examining multiple pathogens simultaneously before making generalizations about infection and spatial patterns. It is imperative to note that these novel findings would have been missed if we had employed the standard univariate analysis used in previous studies. Our results are discussed in the context of disease epidemiology and vector control.
Impact of Malic Enzymes on Antibiotic and Triacylglycerol Production in Streptomyces coelicolor
Authors: Rodriguez Eduardo , Navone Laura , Casati Paula , Gramajo Hugo
Published: April 2012

In this paper, we have characterized two malic enzymes (ME), SCO2951 and SCO5261, fromStreptomyces coelicolorand analyzed their role in antibiotic and triacylglycerol (TAG) production. Biochemical studies have demonstrated that Sco2951 and Sco5261 genes encode NAD+- and NADP+-dependent malic enzymes, respectively. Single or double mutants in the ME-encoding genes show no effect on growth rate compared to the parental M145 strain. However, the single Sco2951 and the double Sco2951 Sco5261 mutants display a strong reduction in the production of the polyketide antibiotic actinorhodin; additionally, the Sco2951 Sco5261 mutant shows a decrease in stored TAGs during exponential growth. The lower production of actinorhodin in the double mutant occurs as a consequence of a decrease in the expression ofactII-ORF4, the transcriptional activator of the actinorhodin gene cluster. On the other hand, the reduced TAG accumulation is not due to reduced transcript levels of fatty acid biosynthetic genes nor to changes in the amount of the precursor acetyl coenzyme A (acetyl-CoA). This mutant accumulates intermediates of the tricarboxylic acid (TCA) cycle that could alter the regulation of the actinorhodin biosynthetic pathway, suggesting that MEs are important anaplerotic enzymes that redirect C4 intermediates from the TCA cycle to maintain secondary metabolism and TAG production inStreptomyces.
Polyphyly of Gut Symbionts in Stinkbugs of the Family Cydnidae
Authors: Hosokawa Takahiro , Kikuchi Yoshitomo , Nikoh Naruo , Fukatsu Takema
Published: April 2012

Symbiotic bacteria associated with midgut crypts of stinkbugs of the family Cydnidae, representing seven species and 13 populations, were investigated. All of the symbionts were species specific, and constituted at least four distinct lineages in the
Gammaproteobacteria
, indicating multiple evolutionary origins of the gut symbionts among the burrower bugs.
A 50-Kilodalton Cry2A Peptide Is Lethal to Bombyx mori and Lymantria dispar
Authors: Ohsawa Masataka , Tanaka Miki , Moriyama Kenta , Shimazu Mitsuaki , Asano Shin-ichiro , Miyamoto Kazuhisa , Haginoya Kohsuke , Mitsui Toshiaki , Kouya Tomoaki , Taniguchi Masayuki , Hori Hidetaka
Published: April 2012

The Cry2Aa3 gene was introduced into asporogenic
Bacillus thuringiensis
, and the synthesized protoxin killed
Bombyx mori
and
Lymantria dispar
larvae. Chymotrypsin hydrolyzed the linkages between 49Tyr/Val50 and 145Lys/Ser146 in the protoxin, and 50- and 58-kDa fragments were generated, respectively. Both peptides killed the larvae of both insects.
The Wood Rot Ascomycete Xylaria polymorpha Produces a Novel GH78 Glycoside Hydrolase That Exhibits α-
l
-Rhamnosidase and Feruloyl Esterase Activities and Releases Hydroxycinnamic Acids from Lignocelluloses
Authors: Nghi Do Huu , Bittner Britta , Kellner Harald , Jehmlich Nico , Ullrich René , Pecyna Marek J. , Nousiainen Paula , Sipilä Jussi , Huong Le Mai , Hofrichter Martin , Liers Christiane
Published: April 2012

Soft rot (type II) fungi belonging to the family Xylariaceae are known to substantially degrade hardwood by means of their poorly understood lignocellulolytic system, which comprises various hydrolases, including feruloyl esterases and laccase. In the present study, several members of the Xylariaceae were found to exhibit high feruloyl esterase activity during growth on lignocellulosic materials such as wheat straw (up to 1,675 mU g
−1
) or beech wood (up to 80 mU g
−1
). Following the ester-cleaving activity toward methyl ferulate, a hydrolase of
Xylaria polymorpha
was produced in solid-state culture on wheat straw and purified by different steps of anion-exchange and size-exclusion chromatography to apparent homogeneity (specific activity, 2.2 U mg
−1
). The peptide sequence of the purified protein deduced from the gene sequence and verified by
de novo
peptide sequencing shows high similarity to putative α-
l
-rhamnosidase sequences belonging to the glycoside hydrolase family 78 (GH78; classified under EC 3.2.1.40). The purified enzyme (98 kDa by SDS-PAGE, 103 kDa by size-exclusion chromatography; pI 3.7) converted diverse glycosides (e.g., α-
l
-rhamnopyranoside and α-
l
-arabinofuranoside) but also natural and synthetic esters (e.g., chlorogenic acid, hydroxycinnamic acid glycoside esters, veratric acid esters, or
p
-nitrophenyl acetate) and released free hydroxycinnamic acids (ferulic and coumaric acid) from arabinoxylan and milled wheat straw. These catalytic properties strongly suggest that
X. polymorpha
GH78 is a multifunctional enzyme. It is the first fungal enzyme that combines glycosyl hydrolase with esterase activities and may help this soft rot fungus to degrade lignocelluloses.
Cognitive interventions to reduce diagnostic error: a narrative review
Authors: Graber Mark L , Kissam Stephanie , Payne Velma L , Meyer Ashley N D , Sorensen Asta , Lenfestey Nancy , Tant Elizabeth , Henriksen Kerm , LaBresh Kenneth , Singh Hardeep
Published: April 2012

Background

Errors in clinical reasoning occur in most cases in which the diagnosis is missed, delayed or wrong. The goal of this review was to identify interventions that might reduce the likelihood of these cognitive errors.

Design

We searched PubMed and other medical and non-medical databases and identified additional literature through references from the initial data set and suggestions from subject matter experts. Articles were included if they either suggested a possible intervention or formally evaluated an intervention and excluded if they focused solely on improving diagnostic tests or provider satisfaction.

Results

We identified 141 articles for full review, 42 reporting tested interventions to reduce the likelihood of cognitive errors, 100 containing suggestions, and one article with both suggested and tested interventions. Articles were classified into three categories: (1) Interventions to improve knowledge and experience, such as simulation-based training, improved feedback and education focused on a single disease; (2) Interventions to improve clinical reasoning and decision-making skills, such as reflective practice and active metacognitive review; and (3) Interventions that provide cognitive ‘help’ that included use of electronic records and integrated decision support, informaticians and facilitating access to information, second opinions and specialists.

Conclusions

We identified a wide range of possible approaches to reduce cognitive errors in diagnosis. Not all the suggestions have been tested, and of those that have, the evaluations typically involved trainees in artificial settings, making it difficult to extrapolate the results to actual practice. Future progress in this area will require methodological refinements in outcome evaluation and rigorously evaluating interventions already suggested, many of which are well conceptualised and widely endorsed.
What factors influence emergency department staff attitudes towards using information technology?
Authors: Ayatollahi Haleh , Bath Peter A , Goodacre Steve , Lo Su Yung , Draegebo Morten , Khan Faheem A
Published: April 2012

Objectives

Information technology (IT) has an important role in the emergency department (ED) functioning, but staff attitudes can influence the way IT is used. Qualitative research into the perceptions of the ED staff has identified a variety of individual, environmental and system factors that may influence attitudes towards using IT. The authors aimed to determine which factors predict attitudes towards using IT and which factors are the most influential.

Methods

Findings from a previous qualitative study were used to develop a self-administered questionnaire measuring individual, environmental and system factors, along with staff attitudes towards using IT. The questionnaire was sent to 535 staff working in three English EDs. Simple linear regression was used to examine the relationship between each potential predictor and user attitude, and multiple regression was used to identify the most important predictors.

Results

Completed questionnaires were returned by 362/535 participants (68%). The factors with the strongest positive association with staff attitudes towards using IT were the perceived individual impact of technology (r2=39%, p<0.001), perceived usefulness (r2=7%, p<0.001), perceived ease of use (r2=2%, p=0.006), perceived subjective norms (r2=1%, p=0.013) and computer experience (r2=1%, p=0.034).

Conclusion

The perceived individual impact of technology is the most important factor in determining ED staff attitude towards using IT. The ED staff are more likely to view using IT systems positively if they can see direct individual benefits arising from their use.
Patients' and emergency clinicians' perceptions of improving pre-hospital pain management: a qualitative study
Authors: Iqbal Mohammad , Spaight Peggy Anne , Siriwardena A Niroshan
Published: April 2012

Background

The authors aimed to investigate patients' and practitioners' views and experiences of pre-hospital pain management to inform improvements in care and a patient-centred approach to treatment.

Methods

This was a qualitative study involving a single emergency medical system. Data were gathered through focus groups and semi-structured interviews. Participants were purposively sampled from patients transported by ambulance to hospital with a painful condition during the past 6 months, ambulance service and emergency department (ED) clinicians. Interviews were audiotaped, transcribed and thematic analysis was conducted.

Results

55 participants were interviewed: 17 patients, 25 ambulance clinicians and 13 ED clinicians. Key themes included: (1) consider beliefs of patients and staff in pain management; (2) widen pain assessment strategies; (3) optimise non-drug treatment; (4) increase drug treatment options; and (5) enhance communication and coordination along the pre-hospital pain management pathway. Patients and staff expected pain to be relieved in the ambulance; however, refusal of or inadequate analgesia were common. Pain was commonly assessed using a verbal score, but practitioners' views of severity were sometimes discordant with this. Morphine and Entonox were commonly used to treat pain. Reassurance, positioning and immobilisation were used as alternatives to drugs. Pre-hospital pain management could be improved by addressing practitioner and patient barriers, increasing available drugs and developing multi-organisational pain management protocols supported by training for staff.

Conclusions

Pain is often poorly managed and undertreated in the pre-hospital environment. The authors' findings may be used to inform guidance, education and policy to improve the pre-hospital pain management pathway.
A randomised trial of granulocyte-macrophage colony-stimulating factor for neonatal sepsis: outcomes at 2 years
Authors: Marlow Neil , Morris Timothy , Brocklehurst Peter , Carr Robert , Cowan Frances M , Patel Nishma , Petrou Stavros , Redshaw Maggie E , Modi Neena , Dore Caroline
Published: April 2012

Objective

The authors performed a randomised trial in very preterm small-for-gestational age (SGA) babies to determine if prophylaxis with granulocyte-macrophage colony-stimulating factor (GM-CSF) improves outcomes (the PROGRAMS trial). Despite increased neutrophil counts following GM-CSF, the authors reported no significant difference in neonatal sepsis-free survival.

Patients and methods

280 babies born <31 weeks of gestation and SGA were entered into the trial. Outcome was determined at 2 years to determine neurodevelopmental and general health outcomes, including economic costs.

Results

The authors found no significant differences in health outcomes or health and social care costs between the trial groups. In the GM-CSF arm, 87 of 134 (65%) babies survived to 2 years without severe disability compared with 87 of 131 (66%) controls (RR: 1·0, 95% CI 0·8 to 1·2). Marginally, more children receiving GM-CSF were reported to have cough (RR 1·7, 95% CI 1·1 to 2·6) and had signs of chronic respiratory disease (Harrison's sulcus; RR 2·0, 95% CI 1·0 to 3·9) though this was not reflected in bronchodilator use or need for hospitalisation for respiratory disease. Overall, the rate of neurologic abnormality (7%–9%) was similar but mean overall developmental scores were lower than expected for gestational age.

Conclusions

The administration of GM-CSF to very preterm SGA babies is not associated with improved or more adverse outcomes at 2 years of age. The apparent excess of developmental impairment in the entire PROGRAMS cohort, without corresponding increase in neurological abnormality, may represent diffuse brain injury attributable to intrauterine growth restriction.
Endoscopic retrograde cholangio-pancreatography services can be accessible and of a high standard in a district general hospital
Authors: Mitra Vikramjit , Mitchison Harriet , Nylander David
Published: April 2012

Background

Endoscopic retrograde cholangio-pancreatography (ERCP) is an important tool in the management of pancreato-biliary disease.

Objective

To compare the current practice of ERCP in a district general hospital with those reported in the 2007 British Society of Gastroenterology (BSG) ERCP audit and assess access to the service.

Design

This was a service evaluation study. Data were collected retrospectively for all people who underwent ERCP. Demographic, clinical and procedure related data were collected and analysed.

Setting

Sunderland Hospital.

Results

236 patients (median age 70 years, 56% women) underwent ERCP. The median period from referral to patient review was 1.0 day. The median period from the decision to carry out an ERCP to the actual procedure date was 3 days. All patients had radiological imaging before their first procedure. 96% patients had their bloods checked within 1 week of the procedure. The most common indication was related to choledocholithiasis and its complications. The mean doses of midazolam and diazemul used were 4.4 mg and 11.1 mg, respectively. The selective biliary cannulation rate was 92%. Sphincterotomy, biliary stent insertion and complete stone extraction were achieved in 94%, 85% and 88% of patients before the procedure. Complications that occurred as a result of ERCPs were as follows: bleeding (1.7%), pancreatitis (3.8%), cholangitis (0.4%) and renal failure (0.4%). The 30-day death rate was 4.6%. However, none of these were procedure related.

Conclusions

The structure of the ERCP services at Sunderland Royal Hospital provides patients with a high-quality and accessible service. The technical success rate and sedation rate were better than those reported in the BSG ERCP audit. The complication rate and procedure-related mortality were comparable to the BSG audit and much below the published figures.