bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in Bacterial Diversity Associated with Epithelial Tissue in the Beef Cow Rumen during the Transition to a High-Grain Diet
Authors: Chen Yanhong , Penner Gregory B. , Li Meiju , Oba Masahito , Guan Le Luo
Published: June 2011

Our understanding of the ruminal epithelial tissue-associated bacterial (defined as epimural bacteria in this study) community is limited. In this study, we aimed to determine whether diet influences the diversity of the epimural bacterial community in the bovine rumen. Twenty-four beef heifers were randomly assigned to either a rapid grain adaptation (RGA) treatment (
n
= 18) in which the heifers were allowed to adapt from a diet containing 97% hay to a diet containing 8% hay over 29 days or to the control group (
n
= 6), which was fed 97% hay. Rumen papillae were collected when the heifers were fed 97%, 25%, and 8% hay diets. PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR analysis were used to characterize rumen epimural bacterial diversity and to estimate the total epimural bacterial population (copy numbers of the 16S rRNA gene). The epimural bacterial diversity from RGA heifers changed (
P
= 0.01) in response to the rapid dietary transition, whereas it was not affected in control heifers. A total of 88 PCR-DGGE bands were detected, and 44 were identified from phyla including
Firmicutes
,
Bacteroidetes
, and
Proteobacteria
. The bacteria
Treponema
sp.,
Ruminobacter
sp., and
Lachnospiraceae
sp. were detected only when heifers were fed 25% and 8% hay diets, suggesting the presence of these bacteria is the result of adaptation to the high-grain diets. In addition, the total estimated population of rumen epimural bacteria was positively correlated with molar proportions of acetate, isobutyrate, and isovalerate, suggesting that they may play a role in volatile fatty acid metabolism in the rumen.
Roles of Two Shewanella oneidensis MR-1 Extracellular Endonucleases
Authors: Gödeke Julia , Heun Magnus , Bubendorfer Sebastian , Paul Kristina , Thormann Kai M.
Published: June 2011

The dissimilatory iron-reducing bacteriumShewanella oneidensisMR-1 is capable of using extracellular DNA (eDNA) as the sole source of carbon, phosphorus, and nitrogen. In addition, we recently demonstrated thatS. oneidensisMR-1 requires eDNA as a structural component during all stages of biofilm formation. In this study, we characterize the roles of twoShewanellaextracellular endonucleases, ExeS and ExeM. While ExeS is likely secreted into the medium, ExeM is predicted to remain associated with the cell envelope. BothexeMandexeSare highly expressed under phosphate-limited conditions. Mutants lackingexeSand/orexeMexhibit decreased eDNA degradation; however, the capability ofS. oneidensisMR-1 to use DNA as the sole source of phosphorus is only affected in mutants lackingexeM. Neither of the two endonucleases alleviates toxic effects of increased eDNA concentrations. The deletion ofexeMand/orexeSsignificantly affects biofilm formation ofS. oneidensisMR-1 under static conditions, and expression ofexeMandexeSdrastically increases during static biofilm formation. Under hydrodynamic conditions, a deletion ofexeMleads to altered biofilms that consist of densely packed structures which are covered by a thick layer of eDNA. Based on these results, we hypothesize that a major role of ExeS and, in particular, ExeM ofS. oneidensisMR-1, is to degrade eDNA as a matrix component during biofilm formation to improve nutrient supply and to enable detachment.
Role of
rpoS
in the Development of Cell Envelope Resilience and Pressure Resistance in Stationary-Phase Escherichia coli
Authors: Charoenwong Duangkamol , Andrews Simon , Mackey Bernard
Published: June 2011

This work investigated the role of
rpoS
in the development of increased cell envelope resilience and enhanced pressure resistance in stationary-phase cells of
Escherichia coli
. Loss of both colony-forming ability and membrane integrity, measured as uptake of propidium iodide (PI), occurred at lower pressures in
E. coli
BW3709 (
rpoS
) than in the parental strain (BW2952). The
rpoS
mutant also released much higher concentrations of protein under pressure than the parent. We propose that RpoS-regulated functions are responsible for the increase in membrane resilience as cells enter stationary phase and that this plays a major role in the development of pressure resistance. Strains from the Keio collection with mutations in two RpoS-regulated genes,
cfa
(cyclopropane fatty acyl phospholipid synthase) and
osmB
(outer membrane lipoprotein), were significantly more pressure sensitive and took up more PI than the parent strain, with
cfa
having the greatest effect. Mutations in the
bolA
morphogene and other RpoS-regulated lipoprotein genes (
osmC
,
osmE
,
osmY
, and
ybaY
) had no effect on pressure resistance. The cytoplasmic membranes of the
rpoS
mutant failed to reseal after pressure treatment, and strains with mutations in
o
smB
and
nlpI
(new lipoprotein) were also somewhat impaired in the ability to reseal their membranes. The
cfa
mutant, though pressure sensitive, was unaffected in membrane resealing, implying that the initial transient permeabilization event is critical for loss of viability rather than the failure to reseal. The enhanced pressure sensitivity of
polA
,
recA
, and
xthA
mutants suggested that DNA may be a target of oxidative stress in pressure-treated cells.
An Essential Tyrosine Phosphatase Homolog Regulates Cell Separation, Outer Membrane Integrity, and Morphology in Caulobacter crescentus
Authors: Shapland Elaine B. , Reisinger Sarah J. , Bajwa Amrita K. , Ryan Kathleen R.
Published: June 2011

Although reversible phosphorylation on tyrosine residues regulates the activity of many eukaryotic proteins, there are few examples of this type of regulation in bacteria. We have identified the first essential tyrosine phosphatase homolog in a bacterium,Caulobacter crescentusCtpA.ctpAmutants with altered active-site residues are nonviable, and depletion of CtpA yields chains of cells with blebbed outer membranes, linked by unresolved peptidoglycan. CtpA overexpression reduces cell curvature in a manner similar to deleting the intermediate filament protein crescentin, but it does not disrupt crescentin localization or membrane attachment. Although it has no obvious signal sequence or transmembrane-spanning domains, CtpA associates with theCaulobacterinner membrane. Immunolocalization experiments suggest that CtpA accumulates at the division site during the last quarter of the cell cycle. We propose that CtpA dephosphorylates one or more proteins involved in peptidoglycan biosynthesis or remodeling, which in turn affect cell separation, cell envelope integrity, and vibrioid morphology.
Genome Sequence of Streptomyces sp. Strain Tü6071
Authors: Erxleben Anika , Wunsch-Palasis Julia , Grüning Björn A. , Luzhetska Marta , Bechthold Andreas , Günther Stefan
Published: June 2011

Streptomyces
sp. Tü6071 is a soil-dwelling bacterium which has a highly active isoprenoid biosynthesis. Isoprenoids are important precursors for biopharmaceutical molecules such as antibiotics or anticancer agents, e.g., landomycin.
Streptomyces
sp. Tü6071 produces the industrially important terpene glycosides phenalinolactones, which have antibacterial activity against several Gram-positive bacteria. The availability of the genome sequence of
Streptomyces
sp. Tü6071 allows for understanding the biosynthesis of these pharmaceutical molecules and will facilitate rational genome modification to improve industrial use.
Draft Genome Sequence of Paenibacillus elgiiB69, a Strain with Broad Antimicrobial Activity
Authors: Ding Rui , Li Yudong , Qian Chaodong , Wu Xuechang
Published: June 2011

Here, we report the draft genome sequence of
Paenibacillus elgii
B69, which was isolated from soil and has broad-spectrum antimicrobial activity. As far as we know, the
P. elgii
genome is the largest of the
Paenibacillus
genus for which genome sequences are available. Multiple sets of genes related to antibiotic biosynthetic pathways have been found in the genome.
Complete Genome Sequence of the Bacterium Porphyromonas gingivalis TDC60, Which Causes Periodontal Disease
Authors: Watanabe Takayasu , Maruyama Fumito , Nozawa Takashi , Aoki Akinobu , Okano Soichiro , Shibata Yasuko , Oshima Kenshiro , Kurokawa Ken , Hattori Masahira , Nakagawa Ichiro , Abiko Yoshimitsu
Published: June 2011

Porphyromonas gingivalis
is a black-pigmented asaccharolytic anaerobe and a major causative agent of periodontitis. Here, we report the complete genome sequence of
P. gingivalis
strain TDC60, which was recently isolated from a severe periodontal lesion in a Japanese patient.
Complete Genome Sequence of Propionibacterium acnesType IB Strain 6609
Authors: Hunyadkürti Judit , Feltóti Zsófia , Horváth Balázs , Nagymihály Marianna , Vörös Andrea , McDowell Andrew , Patrick Sheila , Urbán Edit , Nagy István
Published: June 2011

Propionibacterium acnes
is an anaerobic Gram-positive bacterium that forms part of the normal human cutaneous microbiota and is thought to play a central role in acne vulgaris, a chronic inflammatory disease of the pilosebaceous unit (I. Kurokawa et al., Exp. Dermatol.
18:
821-832, 2009). Here we present the whole genome sequence of
P. acnes
type IB strain 6609, which was recovered from a skin sample from a woman with no recorded acne history and is thus considered a nonpathogenic strain (I. Nagy, Microbes Infect.
8:
2195-2205, 2006).
Genome Sequence of the Thermophilic Strain Bacillus coagulans2-6, an Efficient Producer of High-Optical-Purity
l
-Lactic Acid
Authors: Su Fei , Yu Bo , Sun Jibin , Ou Hong-Yu , Zhao Bo , Wang Limin , Qin Jiayang , Tang Hongzhi , Tao Fei , Jarek Michael , Scharfe Maren , Ma Cuiqing , Ma Yanhe , Xu Ping
Published: June 2011

Bacillus coagulans
2-6 is an efficient producer of lactic acid. The genome of
B. coagulans
2-6 has the smallest genome among the members of the genus
Bacillus
known to date. The frameshift mutation at the start of the
d
-lactate dehydrogenase sequence might be responsible for the production of high-optical-purity
l
-lactic acid.
Genome Analysis of a Mycoplasma hyorhinis Strain Derived from a Primary Human Melanoma Cell Line
Authors: Kornspan Jonathan D. , Lysnyansky Inna , Kahan Tamar , Herrmann Richard , Rottem Shlomo , Nir-Paz Ran
Published: June 2011

The complete genome of
Mycoplasma hyorhinis
strain MCLD has been sequenced and annotated. This genome differs by the inversion of a 14.4-kb and a 3.7-kb fragment and the deletion of a 9.9-kb fragment from
M. hyorhinis
strain HUB-1, isolated from swine respiratory tract. The genome revealed 778 coding sequences (CDSs), with a limited number of
vlp
genes encoding variable surface lipoproteins.
Diffuse reflectance spectroscopy: diagnostic accuracy of a non-invasive screening technique for early detection of malignant changes in the oral cavity
Authors: Jayanthi J L , Nisha G U , Manju S , Philip E K , Jeemon P , Baiju K V , Beena V T , Subhash N
Published: June 2011

Background

Strong proof-of-principle for utilisation of diffuse reflectance spectroscopy, a non-invasive tool for early detection of malignant changes, has emerged recently. The potential of this technique in distinguishing normal tissue from hyperplastic and dysplastic tissues was explored.

Methods

Diffuse reflectance (DR) spectra in the 400–700 nm region were obtained from the buccal mucosa of 96 patients and 34 healthy volunteers. The DR spectral data were compared against the gold standard biopsy and histopathology results. A principal-component analysis was performed for dimensional reduction in the normalised spectral data with linear discriminant analysis as the classifying technique. The receiver operator characteristic curve technique was employed for evaluating the performance of the diagnostic test.

Results

DR spectral features for different lesions, such as normal/healthy, hyperplastic, dysplastic and squamous cell carcinoma (SCC), varied significantly according to the intensity of oxygenated haemoglobin absorption. While the classification based on discriminant scores provided an overall sensitivity of 98.5% and specificity of 96.0% for distinguishing SCC from dysplasia, they were 100.0% and 95.0%, respectively, for distinguishing dysplasia from hyperplasia. Similarly, the analysis yielded a sensitivity of 95.0% and specificity of 100.0% for distinguishing hyperplasia from healthy tissue. The areas under the receiver operator characteristic curves were 0.98 (95% CI 0.95 to 1.00) and 0.95 (95% CI 0.90 to 1.00) for distinguishing dysplasia from SCC and hyperplasia from dysplasia, respectively.

Conclusion

DR spectral data efficiently discriminate healthy tissue from oral malignant lesions. Diagnostic accuracies obtained in this study highlight the potential use of this method for routine clinical practice.
What you need to know when you prescribe a proton pump inhibitor
Authors: Norman A , Hawkey C J
Published: June 2011

Ever since they were launched, proton pump inhibitors (PPIs) have been regarded as profligate prescription interventions and have become a favourite target for pharmacy advisers. Now that they are cheap, with generic omeprazole 20 mg daily costing £1.88 per month (£24.51 per annum) in the UK, it is time to ask whether this status should be reviewed, whether there are areas where the message should be reversed and whether there are any circumstances in which the extra cost of branded PPIs or combined preparations is justified. Equally, with the recognition of an extended toxicity profile, is prescribing profligacy not an economic but a safety issue?
Quantitative expression study of four cytokeratins and p63 in squamous cell carcinoma of the tongue: suitability for sentinel node navigation surgery using one-step nucleic acid amplification
Authors: Yamauchi Kohichi , Fujioka Yasunori , Kogashiwa Yasunao , Kohno Naoyuki
Published: June 2011

Aims

Sentinel node navigation surgery (SNNS) is currently considered to provide better staging of regional metastasis. For rapid and accurate sentinel lymph node analysis, one-step nucleic acid amplification using cytokeratin 19 (CK19) has been applied, particularly in breast cancer. On the other hand, additional quantitative reverse transcription PCR targets containing cytokeratins have been reported recently in head and neck cancer. In this report, CK19 and p63 were immunohistochemically examined in primary tumours for use as molecular markers and were compared with cytokeratin 903 (CK903), cytokeratin 8/18 (CK8/18) and cytokeratin (AE1/AE3), which are used in diagnostic immunohistochemistry for head and neck squamous cell carcinoma.

Methods

The study reviewed 17 patients with T1/T2, N0 (UICC) oral squamous cell carcinoma of the tongue who were treated surgically at Kyorin University Hospital between 2002 and 2009. The intensity and proportion of tumour cells stained for CK19, CK903, p63, CK8/18 and AE1/AE3 were evaluated.

Results

CK19 and CK8/18 staining in cytoplasm was patchy among carcinoma cells, indicating weak expression. Staining proportion for p63, CK903 and AE1/AE3 was greater than for CK19 and CK8/18, although staining intensity for CK903 was weaker than for p63 and AE1/AE3. The difference in total score between CK19 and CK8/18 staining and p63, CK903 and AE1/AE3 staining was statistically significant (p<0.001). p63 and AE1/AE3 may be better markers than CK903, CK19 and CK8/18.

Conclusions

This suggests that p63 is of clinical utility in SNNS and that CK19 is unsuitable for early tongue carcinoma. Further studies are needed before clinical application of these markers.
Sensitive troponin assays
Authors: Collinson Paul O
Published: June 2011

Sensitive troponin assays have been developed to meet the diagnostic goals set by the universal definition of myocardial infarction (MI). The analytical advantages of sensitive troponin assays include improved analytical imprecision at concentrations below the 99th percentile and the ability to define a reference distribution fully. Clinically, the improved sensitivity translates into the ability to diagnosis MI earlier, possibly within 3 h from admission and the ability to use the rate of change of troponin (Δ troponin) for diagnosis. Very sensitive assays may, in appropriately selected populations (perhaps with the addition of Δ troponin), allow diagnosis on hospital admission or within 1–2 h of admission. An elevated troponin level occurring in patients without suspected acute coronary syndromes has, in all studies to date in which outcome has been examined, been shown to indicate an adverse prognosis whatever the underlying clinical diagnosis. Failure of elevation means a good prognosis allowing early, safe hospital discharge, whereas a raised value requires investigation and should help prevent clinically significant pathology being overlooked. Sensitive troponins do present a challenge to the laboratory and the clinician. For the laboratory, the diagnosis of MI requires a change in troponin value. For the clinician, the challenge is to shift from a simplistic yes/no diagnosis of MI based on a single troponin value to a diagnosis that utilises early troponin changes as part of the clinical picture, and to relate the new class of detectable troponin elevation in patients with ischaemic myocardial disease to existing clinical guidelines and trial evidence.
Republished review: Gene therapy for ocular diseases
Authors: Liu Melissa M , Tuo Jingsheng , Chan Chi-Chao
Published: June 2011

The eye is an easily accessible, highly compartmentalised and immune-privileged organ that offers unique advantages as a gene therapy target. Significant advancements have been made in understanding the genetic pathogenesis of ocular diseases, and gene replacement and gene silencing have been implicated as potentially efficacious therapies. Recent improvements have been made in the safety and specificity of vector-based ocular gene transfer methods. Proof-of-concept for vector-based gene therapies has also been established in several experimental models of human ocular diseases. After nearly two decades of ocular gene therapy research, preliminary successes are now being reported in phase 1 clinical trials for the treatment of Leber congenital amaurosis. This review describes current developments and future prospects for ocular gene therapy. Novel methods are being developed to enhance the performance and regulation of recombinant adeno-associated virus- and lentivirus-mediated ocular gene transfer. Gene therapy prospects have advanced for a variety of retinal disorders, including retinitis pigmentosa, retinoschisis, Stargardt disease and age-related macular degeneration. Advances have also been made using experimental models for non-retinal diseases, such as uveitis and glaucoma. These methodological advancements are critical for the implementation of additional gene-based therapies for human ocular diseases in the near future.
PKCα mediates acetylcholine-induced activation of TRPV4-dependent calcium influx in endothelial cells
Authors: Adapala Ravi K. , Talasila Phani K. , Bratz Ian N. , Zhang David X. , Suzuki Makoto , Meszaros J. Gary , Thodeti Charles K.
Published: June 2011

Transient receptor potential vanilloid channel 4 (TRPV4) is a polymodally activated nonselective cationic channel implicated in the regulation of vasodilation and hypertension. We and others have recently shown that cyclic stretch and shear stress activate TRPV4-mediated calcium influx in endothelial cells (EC). In addition to the mechanical forces, acetylcholine (ACh) was shown to activate TRPV4-mediated calcium influx in endothelial cells, which is important for nitric oxide-dependent vasodilation. However, the molecular mechanism through which ACh activates TRPV4 is not known. Here, we show that ACh-induced calcium influx and endothelial nitric oxide synthase (eNOS) phosphorylation but not calcium release from intracellular stores is inhibited by a specific TRPV4 antagonist, AB-159908. Importantly, activation of store-operated calcium influx was not altered in the TRPV4 null EC, suggesting that TRPV4-dependent calcium influx is mediated through a receptor-operated pathway. Furthermore, we found that ACh treatment activated protein kinase C (PKC) α, and inhibition of PKCα activity by the specific inhibitor Go-6976, or expression of a kinase-dead mutant of PKCα but not PKCε or downregulation of PKCα expression by chronic 12- O-tetradecanoylphorbol-13-acetate treatment, completely abolished ACh-induced calcium influx. Finally, we found that ACh-induced vasodilation was inhibited by the PKCα inhibitor Go-6976 in small mesenteric arteries from wild-type mice, but not in TRPV4 null mice. Taken together, these findings demonstrate, for the first time, that a specific isoform of PKC, PKCα, mediates agonist-induced receptor-mediated TRPV4 activation in endothelial cells.
Chronic renal failure alters endothelial function in cerebral circulation in mice
Authors: Bugnicourt Jean-Marc , Silveira Charlène Da , Bengrine Abderrahmane , Godefroy Olivier , Baumbach Gary , Sevestre Henri , Bode-Boeger Stefanie M. , Kielstein Jan T. , Massy Ziad A. , Chillon Jean-Marc
Published: June 2011

We examined structure, composition, and endothelial function in cerebral arterioles after 4 wk of chronic renal failure (CRF) in a well-defined murine model (C57BL/6J and apolipoprotein E knockout female mice). We also determined quantitative expression of endothelial nitric oxide synthase (eNOS), phosphorylated eNOS (on serine 1177 and threonine 495), and caveolin-1; quantitative expression of markers of vascular inflammation or oxidative stress [Rock-1, Rock-2, VCAM-1, and peroxisome proliferator-activated receptor-γ (PPARγ)]; and the plasma concentration of l-arginine and asymmetric dimethylarginine (ADMA). Our hypothesis was that endothelial function would be impaired in cerebral arterioles during CRF following either a decrease in NO production (through alteration of eNOS expression or regulation) or an increase in NO degradation (due to oxidative stress or vascular inflammation). Endothelium-dependent relaxation was impaired during CRF, but endothelium-independent relaxation was not. CRF had no effect on cerebral arteriolar structure and composition. Quantitative expressions of eNOS, eNOS phosphorylated on serine 1177, caveolin-1, Rock-1, Rock-2, and VCAM-1 were similar in CRF and non-CRF mice. In contrast, quantitative expression of PPARγ (which exercises a protective role on blood vessels) was significantly lower in CRF mice, whereas quantitative expression of eNOS phosphorylated on the threonine 495 (the inactive form of eNOS) was significantly higher. Lastly, the plasma concentration of ADMA (a uremic toxin and an endogenous inhibitor of eNOS) was elevated and plasma concentration of l-arginine was low in CRF. In conclusion, endothelial function is impaired in a mouse model of early stage CRF. These alterations may be related (at least in part) to a decrease in NO production.
Identification of the MEK1(F129L) Activating Mutation as a Potential Mechanism of Acquired Resistance to MEK Inhibition in Human Cancers Carrying the B-RafV600E Mutation
Authors: Wang Huisheng , Daouti Sherif , Li Wen-hui , Wen Yang , Rizzo Christine , Higgins Brian , Packman Kathryn , Rosen Neal , Boylan John F. , Heimbrook David , Niu Huifeng
Published: June 2011

Although targeting the Ras/Raf/MEK pathway remains a promising anticancer strategy, mitogen-activated protein/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitors in clinical development are likely to be limited in their ability to produce durable clinical responses due to the emergence of acquired drug resistance. To identify potential mechanisms of such resistance, we established MEK inhibitor–resistant clones of human HT-29 colon cancer cells (HT-29R cells) that harbor the B-RafV600E mutation. HT-29R cells were specifically resistant to MEK inhibition in vitro and in vivo, with drug-induced elevation of MEK/ERK and their downstream targets primarily accountable for drug resistance. We identified MEK1(F129L) mutation as a molecular mechanism responsible for MEK/ERK pathway activation. In an isogenic cell system that extended these findings into other cancer cell lines, the MEK1(F129L) mutant exhibited higher intrinsic kinase activity than wild-type MEK1 [MEK1(WT)], leading to potent activation of ERK and downstream targets. The MEK1(F129L) mutation also strengthened binding to c-Raf, suggesting an underlying mechanism of higher intrinsic kinase activity. Notably, the combined use of Raf and MEK inhibitors overcame the observed drug resistance and exhibited greater synergy in HT-29R cells than the drug-sensitive HT-29 parental cells. Overall, our findings suggested that mutations in MEK1 can lead to acquired resistance in patients treated with MEK inhibitors and that a combined inhibition of Raf and MEK may be potentially useful as a strategy to bypass or prevent drug resistance in the clinic. Cancer Res; 71(16); 5535–45. ©2011 AACR.
TLX1-Induced T-cell Acute Lymphoblastic Leukemia
Authors: De Keersmaecker Kim , Ferrando Adolfo A.
Published: June 2011

The TLX1 transcription factor oncogene is frequently activated by chromosomal translocations in T-cell acute lymphoblastic leukemia (T-ALL) and defines a distinct molecular group of tumors characterized by differentiation arrest at the early cortical stage of thymocyte differentiation and excellent response to therapy. Recent developments from the analysis of genomic data on TLX1-specific transcriptional targets and analysis of the molecular mechanisms of TLX1 transformation in human- and mouse-induced leukemias have shown novel insight into the activity of this transcription factor oncogene. Aberrant expression of TLX1 in T-cell progenitors disrupts normal T-cell development and triggers the development of aneuploidy during T-cell transformation. Importantly, the disruption of the mitotic checkpoint in TLX1-induced tumors may be linked not only to the acquisition of secondary genetic alterations in T-ALL but also to increased sensitivity of these tumors to chemotherapy with drugs targeting the formation of the mitotic spindle. Clin Cancer Res; 17(20); 6381–6. ©2011 AACR.
The Two Faces of Interferon-γ in Cancer
Authors: Zaidi M. Raza , Merlino Glenn
Published: June 2011

Interferon-γ is a cytokine whose biological activity is conventionally associated with cytostatic/cytotoxic and antitumor mechanisms during cell-mediated adaptive immune response. It has been used clinically to treat a variety of malignancies, albeit with mixed results and side effects that can be severe. Despite ample evidence implicating a role for IFN-γ in tumor immune surveillance, a steady flow of reports has suggested that it may also have protumorigenic effects under certain circumstances. We propose that, in fact, IFN-γ treatment is a double-edged sword whose anti- and protumorigenic activities are dependent on the cellular, microenvironmental, and/or molecular context. As such, inhibition of the IFN-γ/IFN-γ receptor pathway may prove to be a viable new therapeutic target for a subset of malignancies. Clin Cancer Res; 17(19); 6118–24. ©2011 AACR.
Activating Effects of Chronic Variable Stress in Rats with Different Exploratory Activity: Association with Dopamine D1 Receptor Function in Nucleus Accumbens
Authors: Matrov Denis , Vonk Argo , Herm Laura , Rinken Ago , Harro Jaanus
Published: June 2011

Background/Aims: Rats display persistent behavioural phenotypes of low (LE) versus high (HE) exploratory activity in the exploration box paradigm. LE rats that prefer passive coping strategies show differential dopaminergic activity in the striatum. The main hypothesis of this study was that chronic variable stress (CVS) would have a higher impact on LE rats. Methods: Animals were submitted to a CVS regimen lasting 32 days that was followed by a behavioural test battery. The functional states of their dopamine D1 and D2 receptors were measured in the striatum and nucleus accumbens (NAcc). Cerebral oxidative metabolism was assessed via cytochrome c oxidase histochemistry in 65 brain regions. Results: CVS decreased weight gain, to a higher extent in LE rats, and lowered the sucrose preference after the first week, but habituation to the anhedonic effect had developed by the end of the experiment. CVS did not change the behavioural phenotypes initially assigned. No effect of stress on D2 receptor function was found. Chronically stressed animals exhibited higher levels of social interaction and D1 receptor-mediated cAMP accumulation in the NAcc, but not in the striatum. CVS was associated with higher oxidative metabolism levels in the anteroventral thalamus, median raphe nuclei and central periaqueductal grey matter. These changes after stress did not depend upon the exploratory phenotype. Conclusion: This study revealed changes in brain biochemistry after habituation to CVS that might be implicated in successful adaptation to chronic stress.
Successful Treatment of Amegakaryocytic Thrombocytopenia with Azathioprine
Authors: Chang Hung , Tang Tzung-Chih
Published: June 2011

Acquired amegakaryocytic thrombocytopenia (AAMT) is an entity characterized by severe thrombocytopenia with a significantly reduced number of megakaryocytes in the bone marrow. AAMT is rare and poorly defined. Therefore, standard treatment is not well established. In general, steroids are considered the frontline treatment while anti-thymocyte globulin and cyclosporine are reported to be effective in scattered reports. We report a case of AAMT which was successfully treated with azathioprine 3 mg/kg/day. The clinical bleeding tendency resolved after treatment for 4 weeks and complete remission was documented after 6 weeks. Azathioprine treatment for AAMT is low risk, convenient, and cost-effective. Our successful experience suggests that azathioprine is potentially the treatment of choice after steroid failure.
Unusually Long Survival of a 67-Year-Old Patient with Near-Tetraploid Acute Myeloid Leukemia M0 without Erythroblastic and Megakaryocytic Dysplasia
Authors: Lemež Petr , Klamová Hana , Zemanová Zuzana , Marinov Iuri , Fuchs Ota , Schwarz Jiří , Březinová Jana , Provazníková Dana , Kostečka Arnošt , Marková Jana , Michalová Kyra , Jelínek Jaroslav
Published: June 2011

Patients with near-tetraploid acute myeloid leukemia (NT-AML) typically have poor survival. We present the case of a 67-year-old Caucasian male with NT-AML M0 who had an unusually long first complete remission of 51 months and an overall survival of 80 months. The only characteristic distinguishing him from other previously described patients with NT-AML was the absence of erythroblastic and/or megakaryocytic dysplasia (EMD) at diagnosis. Molecular-genetic testing for AML fusion transcripts associated with a favorable prognosis (PML/RARα,AML1/ETO, and CBFβ/MYH11) were negative, as were other prognostic markers like MLL-PTD,FLT3-ITD, or mutations of FLT3-D835,NPM1, or CEBPA. Expression studies of ERG,MN1, and EVI1 revealed overexpression of ERG only. The absence of EMD may be a useful prognostic/diagnostic feature of this new rare subtype of NT-AML.
Anti-Infective Properties of Lactobacillus fermentum against Staphylococcus aureus and Pseudomonas aeruginosa
Authors: Varma Parvathi , Nisha N. , Dinesh Kavitha R. , Kumar Anil V. , Biswas Raja
Published: June 2011

Surgical wounds and implant-associated Staphylococcus aureus and Pseudomonas aeruginosa infections are often difficult to treat because of limited susceptibility of several of these strains to conventional antibiotics. As a result, there is a constant need for new alternative drugs. The aim of this study was to investigate the antimicrobial properties of Lactobacillus fermentum, a probiotic bacterium, which we have isolated from colonic biopsies. The inhibition of S. aureus and P. aeruginosa growth was evaluated by coincubating with L. fermentum strains. Growth inhibition was tested for several of their clinical isolates using agar well diffusion assays. For biofilm assay S. aureus and P. aeruginosa were grown on the glass slides and in 96-well plates in presence of 2.5 µg/ml culture filtrate of L. fermentum. Biofilms were photographed using confocal microscope or stained with 0.1% crystal violet. Reduction in the cytotoxicity of S. aureus and P. aeruginosa was observed in presence of 2.5 µg/ml L. fermentum-spent media. Using in vitroexperiments, we showed that L. fermentum-secreted compound(s) inhibits the growth, cytotoxicity and biofilm formation of several S. aureus and P. aeruginosa strains. Compound(s) present in the culture supernatant of L. fermentum may have promising applications in treating hospital-acquired infections.
The Effect of Vascular Risk Factors on the Efficacy of Rivastigmine Patch and Capsule Treatment in Alzheimer’s Disease
Authors: Farlow M.R. , Doraiswamy P.M. , Meng X. , Cooke K. , Somogyi M.
Published: June 2011

Background: Vascular risk factors (VRF) may influence response to rivastigmine in Alzheimer’s disease (AD). Methods: AD patients who participated in a randomized, double-blind, placebo-controlled trial of rivastigmine patch and capsule treatment were stratified by baseline VRF status. Treatment response was evaluated using the AD Assessment Scale-cognitive subscale (ADAS-cog), AD Cooperative Study-Clinical Global Impression of Change (ADCS-CGIC) and the AD Cooperative Study-Activities of Daily Living (ADCS-ADL) scale. Results: ADAS-cog scores significantly improved in all rivastigmine-treated patients (p < 0.05 vs. placebo), except 9.5 mg/24 h patch-treated patients with VRF, and were significantly affected by VRF status in the study population as a whole. Significant benefits were seen on the ADCS-ADL in 9.5 mg/24 h patch- and capsule-treated patients with, but not without, VRF. The ADCS-CGIC significantly improved in capsule-treated patients with, and patch-treated patients without VRF. Although non-significant, patients without VRF showed an apparent faster rate of placebo decline. Conclusion: VRF may influence AD progression and response to rivastigmine.
Insulin-Like Growth Factor I Serum Levels Influence Ischemic Stroke Outcome
Authors: De Smedt Ann , Brouns Raf , Uyttenboogaart Maarten , De Raedt Sylvie , Moens Maarten , Wilczak Nadine , Luijckx Gert-Jan , De Keyser Jacques
Published: June 2011

Background and Purpose—

Insulin-like growth factor I (IGF-I) is neuroprotective in animal models of stroke. We investigated whether serum IGF-I levels in patients with acute ischemic stroke influence stroke severity and outcome.

Methods—

Concentrations of IGF-I and IGF binding protein 3 were measured in serum samples obtained within 6 hours after stroke onset from 255 patients who took part in the placebo arm of the United States and Canadian Lubeluzole in Acute Ischemic Stroke Study. Stroke severity was assessed with the National Institutes of Health Stroke Scale. Multivariate analysis was performed to assess the overall shift in modified Rankin Scale score and changes in the National Institutes of Health Stroke Scale score at 3 months. Survival curves were plotted using the Kaplan-Meier method, and the Cox proportional hazard model was used for multivariate analysis to investigate factors influencing survival.

Results—

After controlling for statistically relevant risk factors, subjects with high IGF-I levels or IGF-I/IGF binding protein 3 ratios had a better neurological and functional outcome at 3 months. Baseline stroke severity was not different between high and low IGF-I groups. In contrast to the low IGF-I group, neurological symptoms gradually improved from Day 3 in the high IGF-I group.

Conclusions—

Our results suggest that high serum IGF-I levels just after ischemic stroke onset are associated with neurological recovery and a better functional outcome.
The von Willebrand Inhibitor ARC1779 Reduces Cerebral Embolization After Carotid Endarterectomy
Authors: Markus Hugh S. , McCollum Charles , Imray Chris , Goulder Michael A. , Gilbert Jim , King Alice
Published: June 2011

Background and Purpose—

Inhibition of von Willebrand factor offers a novel approach to prevention of stroke and myocardial ischemia but has not yet been demonstrated to show efficacy on clinically relevant end points. ARC1779 is an aptamer that inhibits the prothrombotic function of von Willebrand factor by binding to the A1 domain of von Willebrand factor and thereby blocking its interaction with glycoprotein. Phase 1 studies suggest it inhibits platelet aggregation with less increase in bleeding than conventional antiplatelet agents. The effect of ARC 1779 on cerebral emboli immediately after carotid endarterectomy was investigated in a randomized clinical trial.

Methods—

Patients undergoing carotid endarterectomy were randomized double-blind to ARC1779 or placebo administered intravenously. Transcranial Doppler recording, to detect cerebral embolic signals, was performed in the first 3 hours postoperatively. The primary end point was time to first embolic signals.

Results—

Thirty-six patients were recruited, 18 in each arm. The Kaplan-Meier median time to first embolic signals was 83.6 minutes for ARC1779 compared with 5.5 minutes for placebo. Using Cox proportional hazards embolic signals occurred statistically significantly later on ARC1779 (
P
=0.007). Reduced embolic signals counts were correlated with inhibition of von Willebrand factor activity (
P
=0.03). Increased perioperative bleeding and anemia were seen with ARC1779.

Conclusions—

von Willebrand factor inhibition reduces thromboembolism in humans. It may play a role in treatment of stroke and myocardial ischemia. The extent to which bleeding complications occur in nonoperated patients needs to be assessed in further studies.

Clinical Trial Registration—

URL:
http://clinicaltrials.gov
. Unique identifier: NCT00742612.
In vivo versus in vitro protein abundance analysis of Shigella dysenteriaetype 1 reveals changes in the expression of proteins involved in virulence, stress and energy metabolism
Authors: Kuntumalla Srilatha , Zhang Quanshun , Braisted John C , Fleischmann Robert D , Peterson Scott N , Donohue-Rolfe Arthur , Tzipori Saul , Pieper Rembert
Published: June 2011

Background

Shigella dysenteriae serotype 1 (SD1) causes the most severe form of epidemic bacillary dysentery. Quantitative proteome profiling of Shigella dysenteriae serotype 1 (SD1) in vitro (derived from LB cell cultures) and in vivo (derived from gnotobiotic piglets) was performed by 2D-LC-MS/MS and APEX, a label-free computationally modified spectral counting methodology.

Results

Overall, 1761 proteins were quantitated at a 5% FDR (false discovery rate), including 1480 and 1505 from in vitro and in vivo samples, respectively. Identification of 350 cytoplasmic membrane and outer membrane (OM) proteins (38% of in silico predicted SD1 membrane proteome) contributed to the most extensive survey of the Shigella membrane proteome reported so far. Differential protein abundance analysis using statistical tests revealed that SD1 cells switched to an anaerobic energy metabolism under in vivo conditions, resulting in an increase in fermentative, propanoate, butanoate and nitrate metabolism. Abundance increases of transcription activators FNR and Nar supported the notion of a switch from aerobic to anaerobic respiration in the host gut environment. High in vivo abundances of proteins involved in acid resistance (GadB, AdiA) and mixed acid fermentation (PflA/PflB) indicated bacterial survival responses to acid stress, while increased abundance of oxidative stress proteins (YfiD/YfiF/SodB) implied that defense mechanisms against oxygen radicals were mobilized. Proteins involved in peptidoglycan turnover (MurB) were increased, while β-barrel OM proteins (OmpA), OM lipoproteins (NlpD), chaperones involved in OM protein folding pathways (YraP, NlpB) and lipopolysaccharide biosynthesis (Imp) were decreased, suggesting unexpected modulations of the outer membrane/peptidoglycan layers in vivo. Several virulence proteins of the Mxi-Spa type III secretion system and invasion plasmid antigens (Ipa proteins) required for invasion of colonic epithelial cells, and release of bacteria into the host cell cytosol were increased in vivo.

Conclusions

Global proteomic profiling of SD1 comparing in vivo vs. in vitro proteomes revealed differential expression of proteins geared towards survival of the pathogen in the host gut environment, including increased abundance of proteins involved in anaerobic energy respiration, acid resistance and virulence. The immunogenic OspC2, OspC3 and IpgA virulence proteins were detected solely under in vivo conditions, lending credence to their candidacy as potential vaccine targets.
Cucurbitacin-I (JSI-124) activates the JNK/c-Jun signaling pathway independent of apoptosis and cell cycle arrest in B Leukemic Cells
Authors: Ishdorj Ganchimeg , Johnston James B , Gibson Spencer B
Published: June 2011

Background

Cucurbitacin-I (JSI-124) is potent inhibitor of JAK/STAT3 signaling pathway and has anti-tumor activity in a variety of cancer including B cell leukemia. However, other molecular targets of JSI-124 beyond the JAK/STAT3 pathway are not fully understood.

Methods

BJAB, I-83, NALM-6 and primary CLL cells were treated with JSI-124 as indicated. Apoptosis was measured using flow cytometry for accumulation of sub-G1 phase cells (indicator of apoptosis) and Annexin V/PI staining. Cell cycle was analyzed by FACS for DNA content of G1 and G2 phases. Changes in phosphorylation and protein expression of p38, Erk1/2, JNK, c-Jun, and XIAP were detected by Western blot analysis. STAT3 and c-Jun genes were knocked out using siRNA transfection. VEGF expression was determined by mRNA and protein levels by RT-PCR and western blotting. Streptavidin Pull-Down Assay was used to determine c-Jun binding to the AP-1 DNA binding site.

Results

Herein, we show that JSI-124 activates c-Jun N-terminal kinase (JNK) and increases both the expression and serine phosphorylation of c-Jun protein in the B leukemic cell lines BJAB, I-83 and NALM-6. JSI-124 also activated MAPK p38 and MAPK Erk1/2 albeit at lower levels than JNK activation. Inhibition of the JNK signaling pathway failed to effect cell cycle arrest or apoptosis induced by JSI-124 but repressed JSI-124 induced c-Jun expression in these leukemia cells. The JNK pathway activation c-Jun leads to transcriptional activation of many genes. Treatment of BJAB, I-83, and NALM-6 cells with JSI-124 lead to an increase of Vascular Endothelial Growth Factor (VEGF) at both the mRNA and protein level. Knockdown of c-Jun expression and inhibition of JNK activation significantly blocked JSI-124 induced VEGF expression. Pretreatment with recombinant VEGF reduced JSI-124 induced apoptosis.

Conclusions

Taken together, our data demonstrates that JSI-124 activates the JNK signaling pathway independent of apoptosis and cell cycle arrest, leading to increased VEGF expression.
Patient education integrated with acupuncture for relief of cancer-related fatigue randomized controlled feasibility study
Authors: Johnston Michael F , Hays Ron D , Subramanian Saskia K , Elashoff Robert M , Axe Eleanor K , Li Jie-Jia , Kim Irene , Vargas Roberto B , Lee Jihey , Yang LuGe , Hui Ka-Kit
Published: June 2011

Background

Cancer-related fatigue (CRF) is a prominent clinical problem. There are calls for multi-modal interventions.

Methods

We assessed the feasibility of delivering patient education integrated with acupuncture for relief of CRF in a pilot randomized controlled trial (RCT) with breast cancer survivors using usual care as control. Social cognitive and integrative medicine theories guided integration of patient education with acupuncture into a coherent treatment protocol. The intervention consisted of two parts. First, patients were taught to improve self-care by optimizing exercise routines, improving nutrition, implementing some additional evidence-based cognitive behavioral techniques such as stress management in four weekly 50-minute sessions. Second, patients received eight weekly 50-minute acupuncture sessions. The pre-specified primary outcome, CRF, was assessed with the Brief Fatigue Inventory (BFI). Secondary outcomes included three dimensions of cognitive impairment assessed with the FACT-COGv2.

Results

Due to difficulties in recruitment, we tried several methods that led to the development of a tailored recruitment strategy: we enlisted oncologists into the core research team and recruited patients completing treatment from oncology waiting rooms. Compared to usual care control, the intervention was associated with a 2.38-point decline in fatigue as measured by the BFI (90% Confidence Interval from 0.586 to 5.014; p <0.10). Outcomes associated with cognitive dysfunction were not statistically significant.

Conclusions

Patient education integrated with acupuncture had a very promising effect that warrants conducting a larger RCT to confirm findings. An effective recruitment strategy will be essential for the successful execution of a larger-scale trial.

Trial registration

NCT00646633
Strengthening systems for communicable disease surveillance: creating a laboratory network in Rwanda
Authors: Kebede Senait , Gatabazi John B , Rugimbanya Pierre , Mukankwiro Therese , Perry Helen N , Alemu Wondimagegnehu , Ndihokubwayo Jean B , Kramer Michael H , Mukabayire Odette
Published: June 2011

Background

The recent emergence of a novel strain of influenza virus with pandemic potential underscores the need for quality surveillance and laboratory services to contribute to the timely detection and confirmation of public health threats. To provide a framework for strengthening disease surveillance and response capacities in African countries, the World Health Organization Regional Headquarters for Africa (AFRO) developed Integrated Disease Surveillance and Response (IDSR) aimed at improving national surveillance and laboratory systems. IDSR emphasizes the linkage of information provided by public health laboratories to the selection of relevant, appropriate and effective public health responses to disease outbreaks.

Methods

We reviewed the development of Rwanda's National Reference Laboratory (NRL) to understand essential structures involved in creating a national public health laboratory network. We reviewed documents describing the NRL's organization and record of test results, conducted site visits, and interviewed health staff in the Ministry of Health and in partner agencies. Findings were developed by organizing thematic categories and grouping examples within them. We purposefully sought to identify success factors as well as challenges inherent in developing a national public health laboratory system.

Results

Among the identified success factors were: a structured governing framework for public health surveillance; political commitment to promote leadership for stronger laboratory capacities in Rwanda; defined roles and responsibilities for each level; coordinated approaches between technical and funding partners; collaboration with external laboratories; and use of performance results in advocacy with national stakeholders. Major challenges involved general infrastructure, human resources, and budgetary constraints.

Conclusions

Rwanda's experience with collaborative partnerships contributed to creation of a functional public health laboratory network.
Recombinant expression and purification of the 2,5-diketocamphane 1,2-monooxygenase from the camphor metabolizing Pseudomonas putida strain NCIMB 10007
Authors: Kadow Maria , Saß Stefan , Schmidt Marlen , Bornscheuer Uwe T
Published: June 2011

Three different Baeyer-Villiger monooxygenases (BVMOs) were reported to be involved in the camphor metabolism by Pseudomonas putida NCIMB 10007. During (+)-camphor degradation, 2,5-diketocamphane is formed serving as substrate for the 2,5-diketocamphane 1,2-monooxygenase. This enzyme is encoded on the CAM plasmid and depends on the cofactors FMN and NADH and hence belongs to the group of type II BVMOs. We have cloned and recombinantly expressed the oxygenating subunit of the 2,5-diketocamphane 1,2-monooxygenase (2,5-DKCMO) in E. coli followed by His-tag-based affinity purification. A range of compounds representing different BVMO substrate classes were then investigated, but only bicyclic ketones were converted by 2,5-DKCMO used as crude cell extract or after purification. Interestingly, also (-)-camphor was oxidized, but conversion was about 3-fold lower compared to (+)-camphor. Moreover, activity of purified 2,5-DKCMO was observed in the absence of an NADH-dehydrogenase subunit.