bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional Characterization of the Quorum Sensing Regulator RsaL in the Plant-Beneficial Strain Pseudomonas putida WCS358
Authors: Rampioni Giordano , Bertani Iris , Pillai Cejoice Ramachandran , Venturi Vittorio , Zennaro Elisabetta , Leoni Livia
Published: November 2011

In many bacteria, quorum sensing (QS) systems rely on a signal receptor and a synthase producing
N
-acyl-homoserine lactone(s) as the signal molecule(s). In some species, the
rsaL
gene, located between the signal receptor and synthase genes, encodes a repressor limiting signal synthase expression and hence signal molecule production. Here we investigate the molecular mechanism of action of the RsaL protein in the plant growth-promoting rhizobacterium
Pseudomonas putida
WCS358 (RsaL
WCS
). In
P. putida
WCS358, RsaL
WCS
displayed a strong repressive effect on the promoter of the QS signal synthase gene,
ppuI
, while it did not repress the same promoter in
Pseudomonas aeruginosa
. DNase I protection assays showed that purified RsaL
WCS
specifically binds to
ppuI
on a DNA region overlapping the predicted σ
70
-binding site, but such protection was observed only at high protein concentrations. Accordingly, electrophoretic mobility shift assays showed that the RsaL
WCS
protein was not able to form stable complexes efficiently with a probe encompassing the
ppuI
promoter, while it formed stable complexes with the promoter of
lasI
, the gene orthologous to
ppuI
in
P. aeruginosa
. This difference seems to be dictated by the lower dyad symmetry of the RsaL
WCS
-binding sequence on the
ppuI
promoter relative to that on the
lasI
promoter. Comparison of the results obtained
in vivo
and
in vitro
suggests that RsaL
WCS
needs a molecular interactor/cofactor specific for
P. putida
WCS358 to repress
ppuI
transcription. We also demonstrate that RsaL
WCS
regulates siderophore-mediated growth limitation of plant pathogens and biofilm formation, two processes relevant for plant growth-promoting activity.
Refining the Roots of the Beewolf-Streptomyces Symbiosis: Antennal Symbionts in the Rare Genus Philanthinus (Hymenoptera, Crabronidae)
Authors: Kaltenpoth Martin , Yildirim Erol , Gürbüz M. Faruk , Herzner Gudrun , Strohm Erhard
Published: November 2011

Insects engage in symbiotic associations with a large diversity of beneficial microorganisms. While the majority of well-studied symbioses have a nutritional basis, several cases are known in which bacteria protect their host from pathogen infestation. Solitary wasps of the genera
Philanthus
and
Trachypus
(beewolves; Hymenoptera, Crabronidae) cultivate the actinomycete “
Candidatus
Streptomyces philanthi” in specialized antennal gland reservoirs. The symbionts are transferred to the larval cocoon, where they provide protection against pathogenic fungi by producing at least nine different antibiotics. Here we investigated the closest relatives of
Philanthus
and
Trachypus
, the rare genus
Philanthinus
, for the presence of antennal gland reservoirs and symbiotic streptomycetes. Molecular analyses identified “
Ca
. Streptomyces philanthi” in reservoirs of
Philanthinus quattuordecimpunctatus
. Phylogenies based on the 16S rRNA gene suggest that
P. quattuordecimpunctatus
may have acquired “
Ca
. Streptomyces philanthi” by horizontal transfer from other beewolf species. In histological sections and three-dimensional reconstructions, the antennal gland reservoirs were found to occupy six antennal segments (as opposed to only five in
Philanthus
and
Trachypus
) and to be structurally less complex than those of the evolutionarily more derived genera of beewolves. The presence of “
Ca
. Streptomyces philanthi” in antennal glands of
Philanthinus
indicates that the symbiosis between beewolves and
Streptomyces
bacteria is much older than previously thought. It probably evolved along the branch leading to the monophyletic tribe Philanthini, as it seems to be confined to the genera
Philanthus
,
Trachypus
, and
Philanthinus
, which together comprise 172 described species of solitary wasps.
Native Valve Bacillus cereus Endocarditis in a Non-Intravenous-Drug-Abusing Patient
Authors: Thomas Benjamin S. , Bankowski Matthew J. , Lau William K. K.
Published: November 2011

Bacillus cereus
is a rare cause of endocarditis, typically associated with intravenous drug abuse, rheumatic heart disease, prosthetic heart valves, pacemakers, or immunodeficiency. We present the first case of native valve
Bacillus cereus
endocarditis with no apparent risk factors. The patient had a fulminant course requiring emergent valve replacement.
Role of Helicobacter pylori Plasticity Region Genes in Development of Gastroduodenal Diseases
Authors: Sugimoto Mitsushige , Watada Masahide , Jung Sung Woo , Graham David Y. , Yamaoka Yoshio
Published: November 2011

The plasticity region of
Helicobacter pylori
is a large chromosomal segment including isolate-specific open reading frames with characteristics of pathogenicity islands. It remains unclear whether genes in the plasticity region play a role in the pathogenesis of gastric mucosal inflammation and gastroduodenal disease. Our aim was to assess the role of selected genes in the plasticity region in relation to risk of
H. pylori
-related disease and the severity of gastric mucosal damage. We used PCR to study the relation of disease outcome and mucosal damage with four genes in the
H. pylori
plasticity region (
jhp0940
,
jhp0945
,
jhp0947
, and
jhp0949
) from isolates obtained from both Western (
n
= 296) and East Asian (
n
= 217) patients. The prevalence of
jhp0945
,
jhp0947
, and
jhp0949
differed significantly between Western and East Asian isolates. In Western isolates, the presence of
jhp0945
was significantly associated with gastric ulcer, duodenal ulcer, and gastric cancer (odds ratios [95% confidence intervals]: 2.27 [1.04 to 4.98], 1.86 [1.03 to 3.34], and 1.92 [1.03 to 3.56], respectively).
jhp0940
-positive Western isolates were significantly associated with absence of gastric ulcer or duodenal ulcer (0.21 [0.05 to 0.94] and 0.31 [0.12 to 0.78], respectively). No significant difference was observed between inflammatory cell infiltration or atrophy and the presence or absence of plasticity region genes. The outcome of
H. pylori
infections varies widely geographically. These data suggest a possible role for difference in the prevalence of plasticity region genes in the geographic variation in
H. pylori
-related diseases.
Value of the Modified Hodge Test for Detection of Emerging Carbapenemases in Enterobacteriaceae
Authors: Girlich Delphine , Poirel Laurent , Nordmann Patrice
Published: November 2011

The modified Hodge test has an excellent sensitivity for detecting enterobacterial isolates producing Ambler class A (KPC) and class D (OXA-48) carbapenemases. Its sensitivity is low for NDM-1 producers (50%) but is increased to 85.7% by adding ZnSO
4
(100 μg/ml) in the culture medium. However, this test has a low specificity and is time-consuming.
Simultaneously Typing Nine Serotypes of Enteroviruses Associated with Hand, Foot, and Mouth Disease by a GeXP Analyzer-Based Multiplex Reverse Transcription-PCR Assay
Authors: Hu Xiumei , Zhang Yong , Zhou Xiaomian , Xu Banglao , Yang Mengjie , Wang Miao , Zhang Chen , Li Jin , Bai Ruyin , Xu Wenbo , Ma Xuejun
Published: November 2011

Hand, foot, and mouth disease (HFMD) is a contagious enteroviral disease occurring primarily in young children and caused by enterovirus 71 (EV71), coxsackievirus A16 (CVA16), and other serotypes of coxsackievirus and echovirus. In this study, a GeXP analyzer-based multiplex reverse transcription (RT)-PCR assay (GeXP assay) consisting of chimeric primer-based PCR amplification with fluorescent labeling and capillary electrophoresis separation was developed to simultaneously identify nine serotypes of enteroviruses associated with HFMD in China, including EV71, CVA16, CVA4, -5, -9, and -10, and CVB1, -3, and -5. The RNAs extracted from cell cultures of viral isolates and synthetic RNAs via
in vitro
transcription were used to analyze the specificity and sensitivity of the assay. The GeXP assay detected as little as 0.03 tissue culture infective dose (TCID
50
) of EV71 and CVA16, 10 copies of panenterovirus, EV71, CVA16, CVB1, and CVB5, and 100 copies of 10 (including panenterovirus) premixed RNA templates. A total of 180 stool specimens collected from HFMD patients and persons suspected of having HFMD were used to evaluate the clinical performance of this assay. In comparison with the results of conventional methods, the sensitivities of the GeXP assay for detection of panenterovirus, EV71, and CVA16 were 98.79% (163/165), 91.67% (44/48), and 91.67% (33/36), respectively, and the specificities were 80.00% (12/15), 98.48% (130/132), and 100% (144/144), respectively. The concordance of typing seven other serotypes of enteroviruses with the results of conventional methods was 92.59% (25/27). In conclusion, the GeXP assay is a rapid, cost-effective, and high-throughput method for typing nine serotypes of HFMD-associated enteroviruses.
Specific Detection of Naturally Occurring Hepatitis C Virus Mutants with Resistance to Telaprevir and Boceprevir (Protease Inhibitors) among Treatment-Naïve Infected Individuals
Authors: Fonseca-Coronado Salvador , Escobar-Gutiérrez Alejandro , Ruiz-Tovar Karina , Cruz-Rivera Mayra Yolanda , Rivera-Osorio Pilar , Vazquez-Pichardo Mauricio , Carpio-Pedroza Juan Carlos , Ruíz-Pacheco Juan Alberto , Cazares Fernando , Vaughan Gilberto
Published: November 2011

The use of telaprevir and boceprevir, both protease inhibitors (PI), as part of the specifically targeted antiviral therapy for hepatitis C (STAT-C) has significantly improved sustained virologic response (SVR) rates. However, different clinical studies have also identified several mutations associated with viral resistance to both PIs. In the absence of selective pressure, drug-resistant hepatitis C virus (HCV) mutants are generally present at low frequency, making mutation detection challenging. Here, we describe a mismatch amplification mutation assay (MAMA) PCR method for the specific detection of naturally occurring drug-resistant HCV mutants. MAMA PCR successfully identified the corresponding HCV variants, while conventional methods such as direct sequencing, endpoint limiting dilution (EPLD), and bacterial cloning were not sensitive enough to detect circulating drug-resistant mutants in clinical specimens. Ultradeep pyrosequencing was used to confirm the presence of the corresponding HCV mutants. In treatment-naïve patients, the frequency of all resistant variants was below 1%. Deep amplicon sequencing allowed a detailed analysis of the structure of the viral population among these patients, showing that the evolution of the NS3 is limited to a rather small sequence space. Monitoring of HCV drug resistance before and during treatment is likely to provide important information for management of patients undergoing anti-HCV therapy.
Chest Wall Abscess Due to Mycobacterium bovis BCG after Intravesical BCG Therapy
Authors: Kanamori Hajime , Isogami Katsuhiko , Hatakeyama Takashi , Saito Hiroo , Shimada Kazuyoshi , Uchiyama Bine , Aso Noboru , Kaku Mitsuo
Published: November 2011

We report a case of chest wall abscess caused by
Mycobacterium bovis
BCG that arose as a complication 1 year after intravesical BCG instillation. We identified
M. bovis
BCG Tokyo 172 in the abscess by PCR-based typing of
Mycobacterium tuberculosis
complex and analysis of variable number of tandem repeats data.
Picornavirus, the Most Common Respiratory Virus Causing Infection among Patients of All Ages Hospitalized with Acute Respiratory Illness
Authors: Atmar Robert L. , Piedra Pedro A. , Patel Shital M. , Greenberg Stephen B. , Couch Robert B. , Glezen W. Paul
Published: November 2011

We evaluated the prevalence of respiratory virus infection (RVI) in 403 illnesses of 364 persons hospitalized over a 2-year period with acute respiratory conditions using virus-specific reverse transcription-PCR (RT-PCR) assays in addition to cell culture and serology. RVIs were identified in >75% of children under 5 years of age and 25 to 37% of adults. The molecular assays doubled the number of infections identified; picornaviruses were the most frequent in patients of all ages, followed by respiratory syncytial virus and influenza viruses.
Rapid Test for Identification of a Highly Transmissible Mycobacterium tuberculosis Beijing Strain of Sub-Saharan Origin
Authors: Millán-Lou María Isabel , Alonso Henar , Gavín Patricia , Hernández-Febles Melissa , Campos-Herrero María Isolina , Copado Rodolfo , Cañas Fernando , Kremer Kristin , Caminero José Antonio , Martín Carlos , Samper Sofía
Published: November 2011

The development of a rapid test to identify
Mycobacterium tuberculosis
Beijing isolates and specifically strain GC1237, coming from a sub-Saharan country, is needed due to its alarming wide spread on Gran Canaria Island (Spain). A rapid test that detects IS
6110
present between
dnaA
and
dnaN
in the Beijing strains and in a specific site for GC1237 (
Rv2180c
) has been developed. This test would be a useful tool in the surveillance of subsequent cases.
Identification and Characterization of an Aspergillus fumigatus “Supermater” Pair
Authors: Sugui Janyce A. , Losada Liliana , Wang Wei , Varga John , Ngamskulrungroj Popchai , Abu-Asab Mones , Chang Yun C. , O’Gorman Céline M. , Wickes Brian L. , Nierman William C. , Dyer Paul S. , Kwon-Chung Kyung J.
Published: November 2011

The mating efficiency of 50
Aspergillus fumigatus
isolates from both clinical and environmental sources was analyzed. Forty isolates completed the sexual cycle in 4 weeks with variable levels of fertility designated high, medium, or low. Two opposite-mating-type strains exhibiting the highest fertility, AFB62 (
MAT1-1
), isolated from a case of invasive aspergillosis, and AFIR928 (
MAT1-2
), isolated from the environment, were chosen as the supermater pair. Single cleistothecia obtained from a cross of the two strains harbored a minimum of 1 × 10
4
ascospores. The viability of ascospores increased with the age of the fruiting body, 17% at 4 weeks and reaching 95% at 20 weeks. AFB62 and AFIR928 were equally virulent in two different murine models, despite differences in their sources. High recombination frequencies were observed when the closely linked genes
alb1
(AFUA_2G17600) and
abr2
(AFUA_2G17530) were used as genetic markers. Comparative genome hybridization analyses revealed that only 86 genes (ca. 0.86% of the genome) are significantly diverged between AFB62 and AFIR928. The high fertility in a relatively short period, combined with a high degree of virulence and a high recombination frequency, demonstrates that the mating pair AFB62 and AFIR928 provides an excellent tool for genetic studies of
A. fumigatus
.

IMPORTANCE
Aspergillus fumigatus
is a heterothallic fungal pathogen that causes life-threatening infections in immunocompromised hosts. Although heterothallism facilitates genetic study via recombinational analysis, previous work showed that a 6-month incubation period is required for the completion of sexual reproduction in this species. Such a long incubation period impedes progress in genetic research. To discover a highly fertile (supermater) pair that can complete the sexual cycle in a considerably shorter period, we screened 50 strains collected from various geographic regions for mating efficiency. We identified a highly virulent pair of supermaters that can be an invaluable tool for genetic study.
Unbalanced Charge Distribution as a Determinant for Dependence of a Subset of Escherichia coli Membrane Proteins on the Membrane Insertase YidC
Authors: Gray Andrew N. , Henderson-Frost Josephine M. , Boyd Dana , Shirafi Shirin , Niki Hironori , Goldberg Marcia B.
Published: November 2011

Membrane proteins are involved in numerous essential cell processes, including transport, gene regulation, motility, and metabolism. To function properly, they must be inserted into the membrane and folded correctly. YidC, an essential protein in
Escherichia coli
with homologues in other bacteria,
Archaea
, mitochondria, and chloroplasts, functions by incompletely understood mechanisms in the insertion and folding of certain membrane proteins. Using a genome-scale approach, we identified 69
E. coli
membrane proteins that, in the absence of YidC, exhibited aberrant localization by microscopy. Further examination of a subset revealed biochemical defects in membrane insertion in the absence of YidC, indicating their dependence on YidC for proper membrane insertion or folding. Membrane proteins possessing an unfavorable distribution of positively charged residues were significantly more likely to depend on YidC for membrane insertion. Correcting the charge distribution of a charge-unbalanced YidC-dependent membrane protein abrogated its requirement for YidC, while perturbing the charge distribution of a charge-balanced YidC-independent membrane protein rendered it YidC dependent, demonstrating that charge distribution can be a necessary and sufficient determinant of YidC dependence. These findings provide insights into a mechanism by which YidC promotes proper membrane protein biogenesis and suggest a critical function of YidC in all organisms and organelles that express it.

IMPORTANCE
Biological membranes are fundamental components of cells, providing barriers that enclose the cell and separate compartments. Proteins inserted into biological membranes serve critical functions in molecular transport, molecular partitioning, and other essential cell processes. The mechanisms involved in the insertion of proteins into membranes, however, are incompletely understood. The YidC protein is critical for the insertion of a subset of proteins into membranes across an evolutionarily wide group of organisms. Here we identify a large group of proteins that depend on YidC for membrane insertion in
Escherichia coli
, and we identify unfavorable distribution of charge as an important determinant of YidC dependence for proper membrane insertion.
Sensor recorded changes in rates of hand washing with soap in response to the media reports of the H1N1 pandemic in Britain
Authors: Fleischman Diana S , Webster Gregory D , Judah Gaby , de Barra Mícheál , Aunger Robert , Curtis Valerie A
Published: November 2011

Objectives

To examine how the frequency of information regarding a real disease threat influences hand washing with soap.

Design and setting

The authors installed wireless devices in highway service station lavatories in England to record the proportion of individuals washing hands with soap from May 2009 to January 2010.

Participants

Participants were users of men's and women's toilets. Combined there was an average of 6800 participant entrances into the lavatories daily.

Primary outcome measure

The primary outcome measure is the proportion of soap usage to the number of entries into the lavatories.

Results

Hand-washing rates were positively related to both H1NI coverage in blogs and the news; however, these relationships were stronger for men than for women.

Conclusions

Hand washing with soap increases proportionally to the frequency of media key words related to H1N1. Women's hand washing was more strongly associated with incidence of media keywords than men's.
Clinical instructors' perception of a faculty development programme promoting postgraduate year-1 (PGY1) residents' ACGME six core competencies: a 2-year study
Authors: Lee Fa-Yauh , Yang Ying-Ying , Hsu Hui-Chi , Chuang Chiao-Lin , Lee Wei-Shin , Chang Ching-Chih , Huang Chia-Chang , Chen Jaw-Wen , Cheng Hao-Min , Jap Tjin-Shing
Published: November 2011

Objective

The six core competencies designated by Accreditation Council for Graduate Medical Education (ACGME) are essential for establishing a patient centre holistic medical system. The authors developed a faculty programme to promote the postgraduate year 1 (PGY1) resident, ACGME six core competencies. The study aims to assess the clinical instructors' perception, attitudes and subjective impression towards the various sessions of the ‘faculty development programme for teaching ACGME competencies.’

Methods

During 2009 and 2010, 134 clinical instructors participated in the programme to establish their ability to teach and assess PGY1residents about ACGME competencies.

Results

The participants in the faculty development programme reported that the skills most often used while teaching were learnt during circuit and itinerant bedside, physical examination teaching, mini-clinical evaluation exercise (mini-CEX) evaluation demonstration, training workshop and videotapes of ‘how to teach ACGME competencies.’ Participants reported that circuit bedside teaching and mini-CEX evaluation demonstrations helped them in the interpersonal and communication skills domain, and that the itinerant teaching demonstrations helped them in the professionalism domain, while physical examination teaching and mini-CEX evaluation demonstrations helped them in the patients' care domain. Both the training workshop and videotape session increase familiarity with teaching and assessing skills. Participants who applied the skills learnt from the faculty development programme the most in their teaching and assessment came from internal medicine departments, were young attending physician and had experience as PGY1clinical instructors.

Conclusions

According to the clinical instructors' response, our faculty development programme effectively increased their familiarity with various teaching and assessment skills needed to teach PGY1residents and ACGME competencies, and these clinical instructors also then subsequently apply these skills.
“Do you really need to ask me that now?”: a self-audit of interruptions to the ‘shop floor’ practice of a UK consultant emergency physician
Authors: Allard Jon , Wyatt Jonathan , Bleakley Alan , Graham Blair
Published: November 2011

Objectives

To map interruptions encountered by a senior physician performing a variety of everyday tasks on an emergency department (ED) ‘shop floor’ in the UK in order to identify tasks most likely to be interrupted, modes of interruption and those interruptions most likely to result in breaks as suspension of the original task.

Methods

A self-observational audit study of interruptions was undertaken by a consultant emergency physician in a medium-sized ED over 25 separate shifts totalling 119 h. The main outcome measures were type and occurrence of interruption in relation to mode of original task. ‘Success’ of interruptions and number of outstanding tasks were also recorded.

Results

718 interruptions were recorded, with an average of 6 per hour. A mean number of 2.44 outstanding tasks were recorded on each occasion of interruption. Verbal advice, telephone calls and interpretations of x-rays were the most common forms of interruption. 498 interruptions (69%) were successful, defined as interruptions that resulted in a task break (over-riding and suspension of the original task). The most successful interruptions were calls to the resuscitation room (95%). Interruptions from electronic telecommunications systems were extensive (33% of total) with success dependent on the type of communication system. Telephone conversations were rarely interrupted (16% compared with a mean of 69%).

Conclusions

Overt electronic communication systems may have a disproportionate impact in determining the likelihood for successful interruptions. Formal consideration of how to prioritise and manage interruptions from various channels could be usefully added to emergency medicine education and training.
Medical humanities as expressive of Western culture
Authors: Hooker Claire , Noonan Estelle
Published: November 2011

In this paper we articulate a growing awareness within the field of the ways in which medical humanities could be deemed expressive of Western cultural values. The authors suggest that medical humanities is culturally limited by a pedagogical and scholarly emphasis on Western cultural artefacts, as well as a tendency to enact an uncritical reliance upon foundational concepts (such as ‘patient’ and ‘experience’) within Western medicine. Both these tendencies within the field, we suggest, are underpinned by a humanistic emphasis on appreciative or receptive encounters with ‘difference’ among patients that may unwittingly contribute to the marginalisation of some patients and healthcare workers. While cultural difference should be acknowledged as a central preoccupation of medical humanities, we argue that the discipline must continue to expand its scholarly and critical engagements with processes of Othering in biomedicine. We suggest that such improvements are necessary in order to reflect the cultural diversification of medical humanities students, and the geographical expansion of the discipline within non-Western and/or non-Anglophone locations.
Vascular parkinsonism: what makes it different?
Authors: Gupta Deepak , Kuruvilla Abraham
Published: November 2011

Vascular parkinsonism (VP) accounts for 2.5–5% of all cases of parkinsonism in various population based and clinical cohort studies. VP develops as a result of ischaemic cerebrovascular disease, so aetiologically it is classified as secondary parkinsonism. It has been variably referred to in the literature as arteriosclerotic parkinsonism, vascular pseudo-parkinsonism, and lower body parkinsonism. The most important consideration while making a diagnosis of VP should be to differentiate VP from Parkinson’s disease (PD) because of prognostic and therapeutic implications. The salient clinical features in VP which differentiate it from PD are presentation with postural instability and falls rather than with upper limb rest tremor or bradykinesia; short shuffling parkinsonian gait in VP is accompanied by a wider base of stance and variable stride length (parkinsonian-ataxic gait), absence of festination, frequent occurrence of pyramidal signs, and early subcortical dementia. In a patient where the clinical features are suggestive of VP the clinical diagnosis can be supported by demonstration of diffuse white matter lesions and/or strategic subcortical infarcts in the MRI of the brain. The therapeutic options in VP are limited to levodopa, and a poor or non-sustained response to levodopa is another differentiating feature between VP and PD.
Role of the renin–angiotensin system in the nandrolone-decanoate-induced attenuation of the Bezold–Jarisch reflex
Authors: Uggere de Andrade Tadeu , Loiola Leonardo Zanoteli , Alcure Samira Merces Nascimento , Medeiros Ana Raquel Santos , Santos Maria Carmen Lopes Ferreira Silva , Moysés Margareth Ribeiro , Abreu Gláucia Rodrigues de , Lenz Dominik , Bissoli Nazaré Souza
Published: November 2011

The androgen nandrolone decanoate (ND) is known to cause cardiovascular abnormalities, such as attenuation of the Bezold–Jarisch Reflex (BJR), cardiac hypertrophy, and elevation of mean arterial pressure (MAP). Futhermore, a relationship between androgens and the renin–angiotensin system (RAS) has been reported. The purpose of this study was to evaluate the influence of RAS on the BJR, cardiac and prostatic hypertrophy, and MAP evoked by ND. For this, male Wistar rats were treated with ND (10 mg·(kg body mass)–1 for 8 weeks; DECA), or vehicle (control animals; CON), or enalapril (10 mg·(kg body mass)–1, daily; CONE), or ND and enalapril (10 mg ND + 10 mg enalapril per kilogram of body mass; DECAE). After 8 weeks of treatment, the BJR was evaluated by bradycardia and hypotensive responses that were elicited by serotonin administration (2–32 µg·(kg body mass)–1). MAP was assessed; cardiac and prostate hypertrophy were determined by the ratio of the tissue mass:body mass, and by histological analysis of the heart. Animals from the DECA group showed prostatic and cardiac hypertrophy, elevation in mean arterial pressure, and an impairment of BJR. Co-treatment with enalapril inhibited these changes. The data from the present study suggest that RAS has an impact on BJR attenuation, cardiac and prostatic hypertrophy, and the elevation in MAP evoked by ND.
Antioxidant, anti-inflammatory, and antiproliferative activities of organic fractions from the Mediterranean brown seaweed Cystoseira sedoides
Authors: Mhadhebi Lamia , Laroche-Clary Audrey , Robert Jacque , Bouraoui Abderrahman
Published: November 2011

The present study was conducted to evaluate the antioxidant, anti-inflammatory, and antiproliferative activities of organic fractions from Cystoseira sedoides (Desfontaines) C. Agardh . Various fractions of C. sedoides (chloroform (F-CHCl3), ethyl acetate (F-AcOEt), and methanol (F-MeOH)) were screened for total phenol content, as well as antioxidant activity, using the stable radical 1,1-diphenyl-2-picrylhydrazyl (DPPH), and assays for determining the reducing power of these fractions. The anti-inflammatory properties of these fractions were assessed using the carrageenan-induced rat paw oedema model. The antiproliferative activity of C. sedoides fractions was evaluated on normal Madin–Darby canine kiney (MDCK), and fibroblast cells and on cancer cell lines (A549, MCF7, and HCT15), using the ability of the cells to metabolically reduce 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) formazan dyes. The F-CHCl3 and F-AcOEt fractions showed significant total phenolic content at 55.09 and 61.30 mg gallic-acid equivalent/g dried sample, respectively. Using the DPPH method, the F-CHCl3 and the F-AcOEt fractions exhibited the strongest radical scavenging activity, with IC50 120 µg/mL for F-CHCl3 and 121 µg/mL for F-AcOEt, which approaches the activity of the powerful antioxidant standard, Trolox (IC50 = 90 µg/mL). The reducing power of the samples was in the following order: F-AcOEt > F-CHCl3  > F-MeOH fraction. The F-CHCl3 and F-AcOEt fractions of C. sedoides tested at different doses (25 and 50 mg/kg, intraperitoneally (i.p)), exhibited a dose-dependent reduction of rat paw oedema. The percentage of inhibition of oedema, 3 h after carrageenan injection, ranged from 67.71% to 73.49% and from 67.74% to 74.58%, for F-CHCl3 and F-AcOEt, respectively. Their effects are comparable with that of lysine acetylsalicylate (300 mg/kg body mass; i.p.), which is used as a reference drug with the ability to inhibit oedema by 66.14%. Our results revealed that the F-CHCl3 and F-AcOEt fractions from C. sedoides showed important antiproliferative properties towards all of the cancer cell lines studied here, as judged by their IC50 values, which ranged from 52.6 to 66.5 µg/mL for A549; 22.4 to 70.2 µg/mL for MCF7, and 250.6 to 255.3 µg/mL for HCT15. Moreover, no visible destruction or alteration of normal cells was observed, even at 500 µg/mL F-CHCl3 or F-AcOEt. These results suggest that C. sedoides fractions might be used as a potential source of natural antioxidant, anti-inflammatory, and antitumor agents. The purification and determination of the chemical structures of the compounds in these active fractions are under investigation. The results could provide a compound(s) with a promising role in future medicines and nutrition, when used either as a drug or a dietary supplement.
UNSUPERVISED FEATURE SELECTION USING INCREMENTAL LEAST SQUARES
Authors: LIU RONG , RALLO ROBERT , COHEN YORAM
Published: November 2011

An unsupervised feature selection method is proposed for analysis of datasets of high dimensionality. The least square error (LSE) of approximating the complete dataset via a reduced feature subset is proposed as the quality measure for feature selection. Guided by the minimization of the LSE, a kernel least squares forward selection algorithm (KLS-FS) is developed that is capable of both linear and non-linear feature selection. An incremental LSE computation is designed to accelerate the selection process and, therefore, enhances the scalability of KLS-FS to high-dimensional datasets. The superiority of the proposed feature selection algorithm, in terms of keeping principal data structures, learning performances in classification and clustering applications, and robustness, is demonstrated using various real-life datasets of different sizes and dimensions.
IMPLEMENTATION OF GIS-BASED MULTICRITERIA DECISION ANALYSIS WITH VB IN ArcGIS
Authors: OZTURK DERYA , BATUK FATMAGUL
Published: November 2011

This article focuses on the integration of multicriteria decision analysis (MCDA) and geographical information systems (GIS) and introduces a tool, GIS–MCDA, written in visual basic in ArcGIS for GIS-based MCDA. The GIS–MCDA deals with raster-based data sets and includes standardization, weighting and decision analysis methods, and sensitivity analysis. Simple additive weighting, weighted product method, technique for order preference by similarity to ideal solution, compromise programming, analytic hierarchy process, and ordered weighted average for decision analysis; ranking, rating, and pairwise comparison for weighting and linear scale transformation for standardization can be applied by using this tool. The maximum score and score range procedures can be used for linear scale transformation. In this article also an application of the GIS–MCDA to determine the flood vulnerability of the South Marmara Basin in Turkey is examined. To check the validity and reliability of the results, the flood vulnerability layer is compared with flood-affected areas.
SIGNED DISTANCED-BASED TOPSIS METHOD FOR MULTIPLE CRITERIA DECISION ANALYSIS BASED ON GENERALIZED INTERVAL-VALUED FUZZY NUMBERS
Authors: CHEN TING-YU
Published: November 2011

The theory of interval valued fuzzy sets is very valuable for modeling impressions of decision makers. In addition, it gives ability to quantify the ambiguous nature of subjective judgments in an easy way. In this paper, by extending the technique for order preference by similarity to ideal solution (TOPSIS), it is proposed a useful method based on generalized interval valued trapezoidal fuzzy numbers (GITrFNs) for solving multiple criteria decision analysis (MCDA) problems. In view of complexity in handling sophisticated data of GITrFNs, this paper employs the concept of signed distances to establish a simple and effective MCDA method based on the main structure of TOPSIS. An algorithm based on TOPSIS method is established to determine the priority order of given alternatives by using properties of signed distances. Finally, the feasibility of the proposed method is illustrated by a practical example of supplier selection.
Expression of muscle anabolic and metabolic factors in mechanically loaded MLO-Y4 osteocytes
Authors: Juffer Petra , Jaspers Richard T. , Lips Paul , Bakker Astrid D. , Klein-Nulend Jenneke
Published: November 2011

Lack of physical activity results in muscle atrophy and bone loss, which can be counteracted by mechanical loading. Similar molecular signaling pathways are involved in the adaptation of muscle and bone mass to mechanical loading. Whether anabolic and metabolic factors regulating muscle mass, i.e., insulin-like growth factor-I isoforms (IGF-I Ea), mechano growth factor (MGF), myostatin, vascular endothelial growth factor (VEGF), or hepatocyte growth factor (HGF), are also produced by osteocytes in bone in response to mechanical loading is largely unknown. Therefore, we investigated whether mechanical loading by pulsating fluid flow (PFF) modulates the mRNA and/or protein levels of muscle anabolic and metabolic factors in MLO-Y4 osteocytes. Unloaded MLO-Y4 osteocytes expressed mRNA of VEGF, HGF, IGF-I Ea, and MGF, but not myostatin. PFF increased mRNA levels of IGF-I Ea (2.1-fold) and MGF (2.0-fold) at a peak shear stress rate of 44Pa/s, but not at 22Pa/s. PFF at 22 Pa/s increased VEGF mRNA levels (1.8- to 2.5-fold) and VEGF protein release (2.0- to 2.9-fold). Inhibition of nitric oxide production decreased (2.0-fold) PFF-induced VEGF protein release. PFF at 22 Pa/s decreased HGF mRNA levels (1.5-fold) but increased HGF protein release (2.3-fold). PFF-induced HGF protein release was nitric oxide dependent. Our data show that mechanically loaded MLO-Y4 osteocytes differentially express anabolic and metabolic factors involved in the adaptive response of muscle to mechanical loading (i.e., IGF-I Ea, MGF, VEGF, and HGF). Similarly to muscle fibers, mechanical loading enhanced expression levels of these growth factors in MLO-Y4 osteocytes. Although in MLO-Y4 osteocytes expression levels of IGF-I Ea and MGF of myostatin were very low or absent, it is known that the activity of osteoblasts and osteoclasts is strongly affected by them. The abundant expression levels of these factors in muscle cells, in combination with low expression in MLO-Y4 osteocytes, provide a possibility that growth factors expressed in muscle could affect signaling in bone cells.
High-fat diets induce changes in hippocampal glutamate metabolism and neurotransmission
Authors: Valladolid-Acebes Ismael , Merino Beatriz , Principato Antonio , Fole Alberto , Barbas Coral , Lorenzo María P. , García Antonia , Del Olmo Nuria , Ruiz-Gayo Mariano , Cano Victoria
Published: November 2011

Obesity and high-fat (HF) diets have a deleterious impact on hippocampal function and lead to impaired synaptic plasticity and learning deficits. Because all of these processes need an adequate glutamatergic transmission, we have hypothesized that nutritional imbalance triggered by these diets might eventually concern glutamate (Glu) neural pathways within the hippocampus. Glu is withdrawn from excitatory synapses by specific uptake mechanisms involving neuronal (EAAT-3) and glial (GLT-1, GLAST) transporters, which regulate the time that synaptically released Glu remains in the extracellular space and, consequently, the duration and location of postsynaptic receptor activation. The goal of the present study was to evaluate in mouse hippocampus the effect of a short-term high-fat dietary treatment on 1) Glu uptake kinetics, 2) the density of Glu carriers and Glu-degrading enzymes, 3) the density of Glu receptor subunits, and 4) synaptic transmission and plasticity. Here, we show that HF diet triggers a 50% decrease of the Michaelis-Menten constant together with a 300% increase of the maximal velocity of the uptake process. Glial Glu carriers GLT-1 and GLAST were upregulated in HF mice (32 and 27%, respectively), whereas Glu-degrading enzymes glutamine synthase and GABA-decarboxilase appeared to be downregulated in these animals. In addition, HF diet hippocampus displayed diminished basal synaptic transmission and hindered NMDA-induced long-term depression (NMDA-LTD). This was coincident with a reduced density of the NR2B subunit of NMDA receptors. All of these results are compatible with the development of leptin resistance within the hippocampus. Our data show that HF diets upregulate mechanisms involved in Glu clearance and simultaneously impair Glu metabolism. Neurochemical changes occur concomitantly with impaired basal synaptic transmission and reduced NMDA-LTD. Taken together, our results suggest that HF diets trigger neurochemical changes, leading to a desensitization of NMDA receptors within the hippocampus, which might account for cognitive deficits.
Macrophages mediate lung inflammation in a mouse model of ischemic acute kidney injury
Authors: Altmann Christopher , Andres-Hernando Ana , McMahan Rachel H. , Ahuja Nilesh , He Zhibin , Rivard Chris J. , Edelstein Charles Louis , Barthel Lea , Janssen William J. , Faubel Sarah
Published: November 2011

Serum IL-6 is increased in acute kidney injury (AKI) and inhibition of IL-6 reduces AKI-mediated lung inflammation. We hypothesized that circulating monocytes produce IL-6 and that alveolar macrophages mediate lung inflammation after AKI via chemokine (CXCL1) production. To investigate systemic and alveolar macrophages in lung injury after AKI, sham operation or 22 min of renal pedicle clamping (AKI) was performed in three experimental settings: 1) systemic macrophage depletion via diphtheria toxin (DT) injection to CD11b-DTR transgenic mice, 2) DT injection to wild-type mice, and 3) alveolar macrophage depletion via intratracheal (IT) liposome-encapsulated clodronate (LEC) administration to wild-type mice. In mice with AKI and systemic macrophage depletion (CD11b-DTR transgenic administered DT) vs. vehicle-treated AKI, blood monocytes and lung interstitial macrophages were reduced, renal function was similar, serum IL-6 was increased, lung inflammation was improved, lung CXCL1 was reduced, and lung capillary leak was increased. In wild-type mice with AKI administered DT vs. vehicle, serum IL-6 was increased. In mice with AKI and alveolar macrophage depletion (IT-LEC) vs. AKI with normal alveolar macrophage content, blood monocytes and lung interstitial macrophages were similar, alveolar macrophages were reduced, renal function was similar, lung inflammation was improved, lung CXCL1 was reduced, and lung capillary leak was increased. In conclusion, administration of DT in AKI is proinflammatory, limiting the use of the DTR-transgenic model to study systemic effects of AKI. Mice with AKI and either systemic mononuclear phagocyte depletion or alveolar macrophage depletion had reduced lung inflammation and lung CXCL1, but increased lung capillary leak; thus, mononuclear phagocytes mediate lung inflammation, but they protect against lung capillary leak after ischemic AKI. Since macrophage activation and chemokine production are key events in the development of acute lung injury (ALI), these data provide further evidence that AKI may cause ALI.
Potent Induction of Tumor Immunity by Combining Tumor Cryoablation with Anti–CTLA-4 Therapy
Authors: Waitz Rebecca , Solomon Stephen B. , Petre Elena N. , Trumble Anne E. , Fassò Marcella , Norton Larry , Allison James P.
Published: November 2011

Thermal ablation to destroy tumor tissue may help activate tumor-specific T cells by elevating the presentation of tumor antigens to the immune system. However, the antitumor activity of these T cells may be restrained by their expression of the inhibitory T-cell coreceptor CTLA-4, the target of the recently U.S. Food and Drug Administration–approved antibody drug ipilumimab. By relieving this restraint, CTLA-4–blocking antibodies such as ipilumimab can promote tumor rejection, but the full scope of their most suitable applications has yet to be fully determined. In this study, we offer a preclinical proof-of-concept in the TRAMP C2 mouse model of prostate cancer that CTLA-4 blockade cooperates with cryoablation of a primary tumor to prevent the outgrowth of secondary tumors seeded by challenge at a distant site. Although growth of secondary tumors was unaffected by cryoablation alone, the combination treatment was sufficient to slow growth or trigger rejection. In addition, secondary tumors were highly infiltrated by CD4+ T cells and CD8+ T cells, and there was a significant increase in the ratio of intratumoral T effector cells to CD4+FoxP3+ T regulatory cells, compared with monotherapy. These findings documented for the first time an effect of this immunotherapeutic intervention on the intratumoral accumulation and systemic expansion of CD8+ T cells specific for the TRAMP C2–specific antigen SPAS-1. Although cryoablation is currently used to treat a targeted tumor nodule, our results suggest that combination therapy with CTLA-4 blockade will augment antitumor immunity and rejection of tumor metastases in this setting. Cancer Res; 72(2); 430–9. ©2011 AACR.
A Novel Method of Transcriptional Response Analysis to Facilitate Drug Repositioning for Cancer Therapy
Authors: Jin Guangxu , Fu Changhe , Zhao Hong , Cui Kemi , Chang Jenny , Wong Stephen T.C.
Published: November 2011

Little research has been done to address the huge opportunities that may exist to reposition existing approved or generic drugs for alternate uses in cancer therapy. In addition, there has been little work on strategies to reposition experimental cancer agents for testing in alternate settings that could shorten their clinical development time. Progress in each area has lagged, in part, because of the lack of systematic methods to define drug off-target effects (OTE) that might affect important cancer cell signaling pathways. In this study, we addressed this critical gap by developing an OTE-based method to repurpose drugs for cancer therapeutics, based on transcriptional responses made in cells before and after drug treatment. Specifically, we defined a new network component called cancer-signaling bridges (CSB) and integrated it with a Bayesian factor regression model (BFRM) to form a new hybrid method termed CSB-BFRM. Proof-of-concept studies were conducted in breast and prostate cancer cells and in promyelocytic leukemia cells. In each system, CSB-BFRM analysis could accurately predict clinical responses to more than 90% of drugs approved by the U.S. Food and Drug Administration and more than 75% of experimental clinical drugs that were tested. Mechanistic investigation of OTEs for several high-ranking drug–dose pairs suggested repositioning opportunities for cancer therapy, based on the ability to enforce retinoblastoma-dependent repression of important E2F-dependent cell-cycle genes. Together, our findings establish new methods to identify opportunities for drug repositioning or to elucidate the mechanisms of action of repositioned drugs. Cancer Res; 72(1); 33–44. ©2011 AACR.
Epithelial and Stromal Developmental Patterns in a Novel Substitute of the Human Skin Generated with Fibrin-Agarose Biomaterials
Authors: Carriel Víctor , Garzón Ingrid , Jiménez Jose-María , Oliveira Ana-Celeste-Ximenes , Arias-Santiago Salvador , Campos Antonio , Sánchez-Quevedo Maria-Carmen , Alaminos Miguel
Published: November 2011

Development of human skin substitutes by tissue engineering may offer new therapeutic alternatives to the use of autologous tissue grafts. For that reason, it is necessary to investigate and develop new biocompatible biomaterials that support the generation of a proper human skin construct. In this study, we generated a novel model of bioengineered human skin substitute using human cells obtained from skin biopsies and fibrin-agarose biomaterials and we evaluated this model both at the ex vivo and the in vivo levels. Once the dermal fibroblasts and the epithelial keratinocytes were isolated and expanded in culture, we used fibrin-agarose scaffolds for the development of a full-thickness human skin construct, which was evaluated after 1, 2, 3 and 4 weeks of development ex vivo. The skin substitutes were then grafted onto immune-deficient nude mice and analyzed at days 10, 20, 30 and 40 postimplantation using transmission electron microscopy, histochemistry and immunofluorescence. The results demonstrated that the fibrin-agarose artificial skin had adequate biocompatibility and proper biomechanical properties. A proper development of both the bioengineered dermis and epidermis was found after 30 days in vivo, although the tissues kept ex vivo and those implanted in the animal model for 10 or 20 days showed lower levels of differentiation. In summary, our model of fibrin-agarose skin equivalent was able to reproduce the structure and histological architecture of the native human skin, especially after long-term in vivo implantation, suggesting that these tissues could reproduce the native skin.