bluuebunny/crossref_metadata_2025 huggingface dataset:
The new TNM classification of lung cancer in practice
Authors: Rami-Porta R. , Giroux D.J. , Goldstraw P.
Published: July 2011

Educational aims

To describe the changes in the 7th edition of the TNM classification of lung cancer

To discuss the clinical implications of the incorporation of the new TNM classification in everyday practice

To emphasise the key points for a proper pathologic classification

The 7th edition of the TNM (tumour, node, metastases) classification of lung cancer incorporates the proposals of the International Association for the Study of Lung Cancer, whose database included more than 100,000 patients from Asia, Australia, Europe, and North America. The changes affect the T and the M components of the classification, and the stage grouping. The N component remained unaltered, although the present descriptors were validated both in the clinical and pathologic settings. This new TNM classification applies to non-small cell lung cancer, small cell lung cancer and, for the first time, to broncho-pulmonary carcinoids. The innovations allow for a better separation of tumours with significantly different prognosis, and imply a more careful determination of tumour size.
Imaging techniques in lung cancer
Authors: De Wever W. , Coolen J. , Verschakelen J.A.
Published: July 2011

Conventional chest radiography, computed tomography (CT), magnetic resonance (MR) and positron emission tomography (PET) are all imaging techniques used for the detection, characterisation, staging and follow-up of lung cancer. The success of CT is related to the fact that very detailed imaging information of the localisation and the extent of the tumour, the presence of enlarged lymph nodes and the presence of metastatic disease can be provided. Integrated PET/CT combines anatomical information from CT with metabolic information from PET. PET/CT is nowadays the best imaging technique for the staging of lung cancer. MR was used for a long time as a problem-solving tool; however, with the development of new application methods, MR becomes a promising imaging modality in the evaluation of lung cancer patients. In this review, the use of these imaging techniques in the evaluation of lung cancer will be discussed.
Construct and Compare Gene Coexpression Networks with DAPfinder and DAPview
Authors: Skinner Jeff , Kotliarov Yuri , Varma Sudhir , Mine Karina L , Yambartsev Anatoly , Simon Richard , Huyen Yentram , Morgun Andrey
Published: July 2011

Background

DAPfinder and DAPview are novel BRB-ArrayTools plug-ins to construct gene coexpression networks and identify significant differences in pairwise gene-gene coexpression between two phenotypes.

Results

Each significant difference in gene-gene association represents a Differentially Associated Pair (DAP). Our tools include several choices of filtering methods, gene-gene association metrics, statistical testing methods and multiple comparison adjustments. Network results are easily displayed in Cytoscape. Analyses of glioma experiments and microarray simulations demonstrate the utility of these tools.

Conclusions

DAPfinder is a new friendly-user tool for reconstruction and comparison of biological networks.
Detection of quantitative trait loci affecting serum cholesterol, LDL, HDL, and triglyceride in pigs
Authors: Uddin Muhammad Jasim , Duy Do Ngoc , Cinar Mehmet Ulas , Tesfaye Dawit , Tholen Ernst , Juengst Heinz , Looft Christian , Schellander Karl
Published: July 2011

Background

Serum lipids are associated with many serious cardiovascular diseases and obesity problems. Many quantitative trait loci (QTL) have been reported in the pig mostly for performance traits but very few for the serum lipid traits. In contrast, remarkable numbers of QTL are mapped for serum lipids in humans and mice. Therefore, the objective of this research was to investigate the chromosomal regions influencing the serum level of the total cholesterol (CT), triglyceride (TG), high density protein cholesterol (HDL) and low density protein cholesterol (LDL) in pigs. For this purpose, a total of 330 animals from a Duroc × Pietrain F2 resource population were phenotyped for serum lipids using ELISA and were genotyped by using 122 microsatellite markers covering all porcine autosomes for QTL study in QTL Express. Blood sampling was performed at approximately 175 days before slaughter of the pig.

Results

Most of the traits were correlated with each other and were influenced by average daily gain, slaughter date and age. A total of 18 QTL including three QTL with imprinting effect were identified on 11 different porcine autosomes. Most of the QTL reached to 5% chromosome-wide (CW) level significance including a QTL at 5% experiment-wide (GW) and a QTL at 1% GW level significance. Of these QTL four were identified for both the CT and LDL and two QTL were identified for both the TG and LDL. Moreover, three chromosomal regions were detected for the HDL/LDL ratio in this study. One QTL for HDL on SSC2 and two QTL for TG on SSC11 and 17 were detected with imprinting effect. The highly significant QTL (1% GW) was detected for LDL at 82 cM on SSC1, whereas significant QTL (5% GW) was identified for HDL/LDL on SSC1 at 87 cM. Chromosomal regions with pleiotropic effects were detected for correlated traits on SSC1, 7 and 12. Most of the QTL identified for serum lipid traits correspond with the previously reported QTL for similar traits in other mammals. Two novel QTL on SSC16 for HDL and HDL/LDL ratio and an imprinted QTL on SSS17 for TG were detected in the pig for the first time.

Conclusion

The newly identified QTL are potentially involved in lipid metabolism. The results of this work shed new light on the genetic background of serum lipid concentrations and these findings will be helpful to identify candidate genes in these QTL regions related to lipid metabolism and serum lipid concentrations in pigs.
Genetic polymorphisms of innate immunity-related inflammatory pathways and their association with factors related to type 2 diabetes
Authors: Arora Paul , Garcia-Bailo Bibiana , Dastani Zari , Brenner Darren , Villegas Andre , Malik Suneil , Spector Timothy D , Richards Brent , El-Sohemy Ahmed , Karmali Mohamed , Badawi Alaa
Published: July 2011

Background

Type 2 diabetes mellitus (T2DM) has been linked to a state of pre-clinical chronic inflammation resulting from abnormalities in the innate immune pathway. Serum levels of pro-inflammatory cytokines and acute-phase proteins, collectively known as 'inflammatory network', are elevated in the pre-, or early, stages of T2DM and increase with disease progression. Genetic variation can affect the innate immune response to certain environmental factors, and may, therefore, determine an individual's lifetime risk of disease.

Methods

We conducted a cross-sectional study in 6,720 subjects from the TwinsUK Registry to evaluate the association between 18 single nucleotide polymorphisms (SNPs) in five genes (TLR4,IL1A,IL6,TNFA, andCRP) along the innate immunity-related inflammatory pathway and biomarkers of predisposition to T2DM [fasting insulin and glucose, HDL- and LDL- cholesterols, triglycerides (TGs), amyloid-A, sensitive C-reactive protein (sCRP) and vitamin D binding protein (VDBP) and body mass index (BMI)].

Results

Of 18 the SNPs examined for their association with nine metabolic phenotypes of interest, six were significantly associated with five metabolic phenotypes (Bonferroni correction,P ≤ 0.0027). Fasting insulin was associated with SNPs inIL6andTNFA, serum HDL-C with variants ofTNFAandCRPand serum sCRP level with SNPs inCRP. Cross-correlation analysis among the different metabolic factors related to risk of T2DM showed several significant associations. For example, BMI was directly correlated with glucose (r = 0.11), insulin (r = 0.15), sCRP (r = 0.23), LDL-C (r = 0.067) and TGs (r = 0.18) but inversely with HDL-C (r = -0.14). sCRP was also positively correlated (P < 0.0001) with insulin (r = 0.17), amyloid-A (r = 0.39), TGs (r = 0.26), and VDBP (r = 0.36) but inversely with HDL-C (r = -0.12).

Conclusion

Genetic variants in the innate immunity pathway and its related inflammatory cascade is associated with some metabolic risk factors for T2DM; an observation that may provide a rationale for further studying their role as biomarkers for disease early risk prediction.
Radiotherapy may improve overall survival of patients with T3/T4 transitional cell carcinoma of the renal pelvis or ureter and delay bladder tumour relapse
Authors: Chen Bing , Zeng Zhao-Chong , Wang Guo-Min , Zhang Li , Lin Zong-Ming , Sun Li-An , Zhu Tong-Yu , Wu Li-Li , Zhang Jian-Ying , Ji Yuan
Published: July 2011

Background

Since transitional cell carcinoma (TCC) of the upper urinary tract is a relatively uncommon malignancy, the role of adjuvant radiotherapy is unknown.

Methods

We treated 133 patients with TCC of the renal pelvis or ureter at our institution between 1998 and 2008. The 67 patients who received external beam radiotherapy (EBRT) following surgery were assigned to the radiation group (RT). The clinical target volume included the renal fossa, the course of the ureter to the entire bladder, and the paracaval and para-aortic lymph nodes, which were at risk of harbouring metastatic disease in 53 patients. The tumour bed or residual tumour was targeted in 14 patients. The median radiation dose administered was 50 Gy. The 66 patients who received intravesical chemotherapy were assigned to the non-radiation group (non-RT).

Results

The overall survival rates for the RT and non-RT groups were not significantly different (p = 0.198). However, there was a significant difference between the survival rates for these groups based on patients with T3/T4 stage cancer. A significant difference was observed in the bladder tumour relapse rate between the irradiated and non-irradiated bladder groups (p = 0.004). Multivariate analysis indicated that improved overall survival was associated with age < 60 years, T1 or T2 stage, absence of synchronous LN metastases, and EBRT. Acute gastrointestinal and bladder reactions were the most common symptoms, but mild non-severe (> grade 3) hematologic symptoms also occurred.

Conclusion

EBRT may improve overall survival for patients with T3/T4 cancer of the renal pelvis or ureter and delay bladder tumour recurrence in all patients.
Bone turnover in passive smoking female rat: relationships to change in bone mineral density
Authors: Gao Shu-guang , Li Kang-hua , Xu Mai , Jiang Wei , Shen Hong , Luo Wei , Xu Wen-shuo , Tian Jian , Lei Guang-hua
Published: July 2011

Background

Many studies have identified smoking as a risk factor for osteoporosis, but it is unclear whether passive smoking has an effect on bone mineral density and bone turnover and if such an effect could cause osteoporosis.The purpose of the study was to investigate the effect of passive smoking on bone mineral density (BMD) and bone turnover and the relationship between BMD and bone turnover in female rat.

Methods

Forty-eight female Wistar rats were randomized into six groups: 2-month, 3-month,4-month smoke-exposed rats and their controls. A rat model of passive cigarette smoking was prepared by breeding female rats in a cigarette-smoking box for 2, 3 or 4 months. Serums were analyzed for levels of osteocalcin, bone-specific alkaline phosphatase (b-ALP) and Tartrate-resistant acid phosphatase 5b (TRACP 5b). BMD was assessed at lumbar vertebrae and femur by dual energy X-ray absorptiometry in passive smoking rats and in control rats.

Results

BMD of lumbar spine and femur was lower in 4-month smoke-exposed female rats than that in controls. However, there was no significant difference in serum osteocalcin levels between smoke-exposed rats and controls. Significantly lower b-ALP and higher TRACP 5b were found in the 3-month or 4-month smoke-exposed rats compared to controls. Subsequent analysis showed that b-ALP positively correlated with BMD of the lumbar vertebrae(r = 0.764, P = 0.027) and femur(r = 0.899, P = 0.002) in 4-month smoke-exposed female rats. Furthermore, TRACP 5b levels negatively correlated with BMD of lumbar vertebrae (r = -0.871, P = 0.005) and femur (r = -0.715, P = 0.046) in 4-month smoke-exposed female rats.

Conclusion

Our data suggest that smoke exposure can inhibit bone formation and increase bone resorption. The hazardous effects of passive smoking on bone status are associated with increased bone turnover in female rat.
Plasmodium vivax: comparison of immunogenicity among proteins expressed in the cell-free systems of Escherichia coli and wheat germ by suspension array assays
Authors: Rui Edmilson , Fernandez-Becerra Carmen , Takeo Satoru , Sanz Sergi , Lacerda Marcus VG , Tsuboi Takafumi , del Portillo Hernando A
Published: July 2011

Background

In vitro cell-free systems for protein expression with extracts from prokaryotic (Escherichia coli) or eukaryotic (wheat germ) cells coupled to solid matrices have offered a valid approach for antigen discovery in malaria research. However, no comparative analysis of both systems is presently available nor the usage of suspension array technologies, which offer nearly solution phase kinetics.

Methods

Five Plasmodium vivax antigens representing leading vaccine candidates were expressed in the E. coli and wheat germ cell-free systems at a 50 μl scale. Products were affinity purified in a single-step and coupled to luminex beads to measure antibody reactivity of human immune sera.

Results

Both systems readily produced detectable proteins; proteins produced in wheat germ, however, were mostly soluble and intact as opposed to proteins produced in E. coli, which remained mostly insoluble and highly degraded. Noticeably, wheat germ proteins were recognized in significantly higher numbers by sera of P. vivax patients than identical proteins produced in E. coli.

Conclusions

The wheat germ cell-free system offers the possibility of expressing soluble P. vivax proteins in a small-scale for antigen discovery and immuno-epidemiological studies using suspension array technology.
Chronologically scheduled snacking with high-protein products within the habitual diet in type-2 diabetes patients leads to a fat mass loss: a longitudinal study
Authors: Navas-Carretero Santiago , Abete Itziar , Zulet M Angeles , Martínez J Alfredo
Published: July 2011

Background

Obesity is the most relevant overnutrition disease worldwide and is associated to different metabolic disorders such as insulin resistance and type-2 diabetes. Low glycemic load foods and diets and moderately high protein intake have been shown to reduce body weight and fat mass, exerting also beneficial effects on LDL-cholesterol, triglyceride concentrations, postprandial glucose curve and HDL-cholesterol levels. The present study aimed at studying the potential functionality of a series of low glycemic index products with moderately high protein content, as possible coadjuvants in the control of type-2 diabetes and weight management following a chronologically planned snacking offer (morning and afternoon).

Methods

The current trial followed a single group, sequential, longitudinal design, with two consecutive periods of 4 weeks each. A total of 17 volunteers participated in the study. The first period was a free living period, with volunteers' habitual ad libitum dietary pattern, while the second period was a free-living period with structured meal replacements at breakfast, morning snack and afternoon snack, which were exchanged by specific products with moderately high protein content and controlled low glycemic index, following a scheduled temporal consumption. Blood extractions were performed at the beginning and at the end of each period (free-living and intervention). Parameters analysed were: fasting glucose, insulin, glycosylated hemoglobin, total-, HDL- and LDL-cholesterol, triglyceride, C - reactive protein and Homocysteine concentrations. Postprandial glucose and insulin were also measured. Anthropometrical parameters were monitored each 2 weeks during the whole study.

Results

A modest but significant (p = 0.002) reduction on body weight (1 kg) was observed during the intervention period, mainly due to the fat mass loss (0.8 kg, p = 0.02). This weight reduction was observed without apparently associated changes in total energy intake. None of the biochemical biomarkers measured was altered throughout the whole study.

Conclusions

Small changes in the habitual dietary recommendations in type-2 diabetes patients by the inclusion of specific low-glycemic, moderately high-protein products in breakfast, morning and afternoon snacks may promote body weight and fat-mass loss, without apparently altering biochemical parameters and cardiovascular risk-related factors.

Trial Registration

Trial registered at clinicaltrials.gov NCT01264523.
Integrin subunits alpha5 and alpha6 regulate cell cycle by modulating the chk1 and Rb/E2F pathways to affect breast cancer metastasis
Authors: Wang Yanfang , Shenouda Sylvia , Baranwal Somesh , Rathinam Rajamani , Jain Prachi , Bao Lili , Hazari Siddhartha , Dash Srikanta , Alahari Suresh K
Published: July 2011

Background

Although integrins have been implicated in the progression of breast cancer, the exact mechanism whereby they exert this regulation is clearly not understood. To understand the role of integrins in breast cancer, we examined the expression levels of several integrins in mouse breast cancer cell lines by flow cytometry and the data were validated by Western and RT-PCR analysis. The importance of integrins in cell migration and cell invasion was examined byin vitroassays. Further the effect of integrins on metastasis was investigated byin vivoexperimental metastasis assays using mouse models.

Results

Integrin α5 subunit is highly expressed in the nonmetastatic cell line 67NR and is significantly low in the highly invasive cell line 4T1. In contrast, expression levels of integrin α6 subunit are high in 4T1 cells and low in 67NR cells.In vitrodata indicated that overexpression of α5 subunit and knockdown of α6 integrin subunit inhibited cell proliferation, migration, and invasion. Ourin vivofindings indicated that overexpression of integrin α5 subunit and knockdown of α6 subunit decreased the pulmonary metastasis property of 4T1 cells. Our data also indicated that overexpression of alpha 5 integrin subunit and suppression of alpha6 integrin subunit inhibited cells entering into S phase by up-regulating p27, which results in downregulation of cyclinE/CDK2 complexes, This suggests that these integrins regulate cell growth through their effects on cell-cycle-regulated proteins. We also found that modulation of these integrins upregulates E2F, which may induce the expression of chk1 to regulate cdc25A/cyclin E/CDK2/Rb in a feedback loop mechanism.

Conclusion

This study indicates that Integrin α5 subunit functions as a potential metastasis suppressor, while α6 subunit functions as a metastasis promoter. The modulation of integrins reduces cdc25 A, another possible mechanism for downregulation of CDK2. Taken together we demonstrate a link between integrins and the chk1-cdc25-cyclin E/CDK2-Rb pathway.
The role of 18F-FDG-PET/CT in the preoperative staging and posttherapy follow up of gastriccancer:Comparison with spiral CT
Authors: Ozkan Elgin , Araz Mine , Soydal Cigdem , Kucuk Ozlem N.
Published: July 2011

Background

The aim of this study was to investigate the role of F-18 fluoro-deoxy-glucose (FDG) positron emission tomography and computed tomography (PET/CT) in the preoperative and posttherapy restaging of gastric cancer and to compare with spiral computerized tomography (CT).

Method

A total of 42 PET/CT scans of 36 gastric cancer patients (28M, 8F; mean age: 56,0 ± 15) were included in the study. A retrospective analysis of the PET/CT results of the patients were compared with concurrent CT results. Confirmation was made by clinical course and serial imaging studies in the follow up. The compatibility ratios were calculated and the accuracy of the PET/CT was assessed. Agreement between PET/CT and concurrent CT was calculated using kappa statistics.

Results

Patients were separated into 3 groups: the patients who were referred to our clinic for preoperative staging (4 patients), for posttherapy evaluation (24 patients) and for the suspicion of local recurrence and/or metastasis exploration after a disease free period (8 patients). Groups 1 and 3 included a small number of patients so they were omitted from the statistical analysis. Focusing on Goup 2, the overall concordance rate was 50% (12 patients). Region based analysis showed the rates of concordance for local recurrence, local lymph node metastasis and distant metastasis were 91% (Kappa: 0.70), 95% (Kappa:0.86) and 50% (Kappa:0.26) respectively. Distant metastases were also investigated in detail and the two techniques showed a concordance of 91% (Kappa: 0.75) for liver, 79%(Kappa:0.31) for distant lymph node, 79% (0.42) for lung, 87%(Kappa:0.33) for bone and 95% for intestinal wall metastasis.

Conclusion

PET/CT is a complementary imaging method which can be successfully used in both preoperative and posttherapy evaluation of gastric cancer.
A novel MCF-10A line allowing conditional oncogene expression in 3D culture
Authors: Herr Ricarda , Wöhrle Franziska U , Danke Christina , Berens Christian , Brummer Tilman
Published: July 2011

Introduction

Non-transformed mammary epithelial cell lines such as MCF-10A recapitulate epithelial morphogenesis in three-dimensional (3D) tissue culture by forming acinar structures. They represent an important tool to characterize the biological properties of oncogenes and to model early carcinogenic events. So far, however, these approaches were restricted to cells with constitutive oncogene expression prior to the set-up of 3D cultures. Although very informative, this experimental setting has precluded the analysis of effects caused by sudden oncoprotein expression or withdrawal in established epithelial cultures. Here, we report the establishment and use of a stable MCF-10A cell line (MCF-10Atet) fitted with a novel and improved doxycycline (dox)-regulated expression system allowing the conditional expression of any transgene.

Methods

MCF-10Atet cells were generated by stable transfection with pWHE644, a vector expressing a second generation tetracycline-regulated transactivator and a novel transcriptional silencer. In order to test the properties of this new repressor/activator switch, MCF-10Atet cells were transfected with a second plasmid, pTET-HABRAF-IRES-GFP, which responds to dox treatment with the production of a bi-cistronic transcript encoding hemagglutinin-tagged B-Raf and green fluorescent protein (GFP). This improved conditional expression system was then characterized in detail in terms of its response to various dox concentrations and exposure times. The plasticity of the phenotype provoked by oncogenic B-RafV600E in MCF-10Atet cells was analyzed in 3D cultures by dox exposure and subsequent wash-out.

Results

MCF-10Atet cells represent a tightly controlled, conditional gene expression system. Using B-RafV600E as a model oncoprotein, we show that its sudden expression in established 3D cultures results in the loss of acinar organization, the induction of an invasive phenotype and hallmarks of epithelial-to-mesenchymal transition (EMT). Importantly, we show for the first time that this severe transformed phenotype can be reversed by dox wash-out and concomitant termination of oncogene expression.

Conclusions

Taken together, we have generated a stable MCF-10A subline allowing tight dox-controlled and reversible expression of any transgene without the need to modify its product by introducing artificial dimerization or ligand-binding domains. This system will be very valuable to address phenomena such as EMT, oncogene addiction, oncogene-induced senescence and drug resistance.
Topological confinement in an antisymmetric potential in bilayer graphene in the presence of a magnetic field
Authors: Zarenia Mohammad , Pereira Joao Milton , Peeters François Maria , de Aquino Farias Gil
Published: July 2011

We investigate the effect of an external magnetic field on the carrier states that are localized at a potential kink and a kink-antikink in bilayer graphene. These chiral states are localized at the interface between two potential regions with opposite signs.

PACS numbers: 71.10.Pm, 73.21.-b, 81.05.Uw
The investigation of frequency response for the magnetic nanoparticulate assembly induced by time-varied magnetic field
Authors: Sun Jianfei , Sui Yunxia , Wang Chunyu , Gu Ning
Published: July 2011

The field-induced assembly of γ-Fe2O3nanoparticles under alternating magnetic field of different frequency was investigated. It was found that the assembly was dependent upon the difference between colloidal relaxation time and field period. The same experiments on DMSA-coated γ-Fe2O3nanoparticles exhibited that the relaxation time may be mainly determined by the magnetic size rather than the physical size. Our results may be valuable for the knowledge of dynamic assembly of colloidal particles.
Phylogenetic and molecular characterization of equine H3N8 influenza viruses from Greece (2003 and 2007): Evidence for reassortment between evolutionary lineages
Authors: Bountouri Maria , Fragkiadaki Eirini , Ntafis Vasileios , Kanellos Theo , Xylouri Eftychia
Published: July 2011

Background

For first time in Greece equine influenza virus infection was confirmed, by isolation and molecular analysis, as the cause of clinical respiratory disease among unvaccinated horses during 2003 and 2007 outbreaks.

Methods

Equine influenza virus (EIV) H3N8 was isolated in MDCK cells from 30 nasal swabs from horses with acute respiratory disease, which were tested positive by Directigen Flu A. Isolation was confirmed by haemagglutination assay and RT-PCR assay of the M, HA and NA gene.

Results

HA sequences of the Greek isolates appeared to be more closely related to viruses isolated in early 1990s in Europe. These results suggested that viruses with fewer changes than those on the main evolutionary lineage may continue to circulate. On the other hand, analysis of deduced NA amino acid sequences were more closely related to viruses isolated in outbreaks in Europe and Asia during 2003-2007. Phylogenetic analysis characterized the Greek isolates as a member of the Eurasian lineage by the haemagglutinin (HA) protein alignment, but appeared to be a member of the Florida sublineage clade 2 by the neuraminidase (NA) protein sequence suggesting that reassortment might be a possible explanation.

Conclusion

Our findings suggest that the Greek strains represent an example of "frozen evolution" and probably reassortment between genetically distinct co-circulated strains. Therefore expanding current equine influenza surveillance efforts is a necessity.
Reduced Spinal Microglial Activation and Neuropathic Pain after Nerve Injury in Mice Lacking all Three Nitric Oxide Synthases
Authors: Kuboyama Kazuya , Tsuda Makoto , Tsutsui Masato , Toyohara Yumiko , Tozaki-Saitoh Hidetoshi , Shimokawa Hiroaki , Yanagihara Nobuyuki , Inoue Kazuhide
Published: July 2011

Background:

Several studies have investigated the involvement of nitric oxide (NO) in acute and chronic pain using mice lacking a single NO synthase (NOS) gene among the three isoforms: neuronal (nNOS), inducible (iNOS) and endothelial (eNOS). However, the precise role of NOS/NO in pain states remains to be determined owing to the substantial compensatory interactions among the NOS isoforms. Therefore, in this study, we used mice lacking all three NOS genes ( n/i/e/NOS−/− mice) and investigated the behavioral phenotypes in a series of acute and chronic pain assays.

Results:

In a model of tissue injury-induced pain, evoked by intraplantar injection of formalin, both iNOS−/− and n/i/eNOS−/− mice exhibited attenuations of pain behaviors in the second phase compared with that in wild-type mice. In a model of neuropathic pain, nerve injury-induced behavioral and cellular responses (tactile allodynia, spinal microglial activation and Src-family kinase phosphorylation) were reduced in n/i/eNOS−/− but not iNOS−/− mice. Tactile allodynia after nerve injury was improved by acute pharmacological inhibition of all NOSs and nNOS. Furthermore, in MG-5 cells (a microglial cell-line), interferon-γ enhanced NOSs and Mac-1 mRNA expression, and the Mac-1 mRNA increase was suppressed by L-NAME co-treatment. Conversely, the NO donor, sodium nitroprusside, markedly increased mRNA expression of Mac-1, interleukin-6, toll-like receptor 4 and P2X4 receptor.

Conclusions:

Our results provide evidence that the NOS/NO pathway contributes to behavioral pain responses evoked by tissue injury and nerve injury. In particular, nNOS may be important for spinal microglial activation and tactile allodynia after nerve injury.
Vascular pathology in multiple sclerosis: mind boosting or myth busting?
Authors: Waschbisch Anne , Manzel Arndt , Linker Ralf A , Lee De-Hyung
Published: July 2011

The investigation of central nervous system vascular changes in the pathophysiology of multiple sclerosis (MS) is a time-honored concept. Yet, recent reports on changes in venous cerebrospinal outflow, the advent of new magnetic resonance imaging techniques and the investigation of immunomodulatory properties of several vascular mediators on the molecular level have added new excitement to hypotheses centering around vascular pathology as determining factor in the pathophysiology of MS. Here we critically review the concept of chronic cerebrospinal venous insufficiency in MS patients and describe new imaging techniques including perfusion weighted imaging, susceptibility weighted imaging and diffusion weighted imaging which reveal central nervous system hypoperfusion, perivascular iron deposition and diffuse structural changes in the MS brain. On a molecular basis, vascular mediators represent interesting targets connecting vascular pathology with immunomodulation. In summary, the relation of venous changes to the pathophysiology of MS may not be as simple as initially described and it certainly seems awkward to think of the complex disease MS solely as result of a simple venous outflow obstruction. Yet, the investigation of new vascular concepts as one variable in the pathophysiology of the autoimmune attack seems very worthwhile and may add to a better understanding of this devastating disorder.
Detection and differentiation of bacterial spores in a mineral matrix by Fourier transform infrared spectroscopy (FTIR) and chemometrical data treatment
Authors: Brandes Ammann Andrea , Brandl Helmut
Published: July 2011

Background

Fourier transform infrared spectroscopy (FTIR) has been used as analytical tool in chemistry for many years. In addition, FTIR can also be applied as a rapid and non-invasive method to detect and identify microorganisms. The specific and fingerprint-like spectra allow - under optimal conditions - discrimination down to the species level. The aim of this study was to develop a fast and reproducible non-molecular method to differentiate pure samples of Bacillus spores originating from different species as well as to identify spores in a simple matrix, such as the clay mineral, bentonite.

Results

We investigated spores from pure cultures of seven different Bacillus species by FTIR in reflection or transmission mode followed by chemometrical data treatment. All species investigated (B. atrophaeus, B. brevis, B. circulans, B. lentus, B. megaterium, B. subtilis, B. thuringiensis) are typical aerobic soil-borne spore formers. Additionally, a solid matrix (bentonite) and mixtures of benonite with spores of B. megaterium at various wt/wt ratios were included in the study. Both hierarchical cluster analysis and principal component analysis of the spectra along with multidimensional scaling allowed the discrimination of different species and spore-matrix-mixtures.

Conclusions

Our results show that FTIR spectroscopy is a fast method for species-level discrimination of Bacillus spores. Spores were still detectable in the presence of the clay mineral bentonite. Even a tenfold excess of bentonite (corresponding to 2.1 × 1010 colony forming units per gram of mineral matrix) still resulted in an unambiguous identification of B. megaterium spores.
Severe Cortisol Deficiency Associated with Reversible Growth Hormone Deficiency in Two Infants: What Is the Link?
Authors: McEachern Rebecca , Drouin Jacques , Metherell Louise , Huot Céline , Van Vliet Guy , Deal Cheri
Published: July 2011

Context:

Hypoglycemia is potentially life-threatening, especially in infants, and can be due to congenital cortisol and/or GH deficiency (GHD).

Case Illustration:

Two full-term infants had undetectable cortisol levels, but also low GH levels, at the time of severe hypoglycemia. GHD persisted for several months, even after cortisol replacement.

Methods:

Targeted molecular investigations were performed and revealed homozygous inactivating mutations in MRAP (MIM ID 609196) in patient 1 and in TPIT (MIM ID 604614) in patient 2. Because GHD is not part of the MRAP or TPIT phenotypes, we performed GH stimulation tests. These revealed that GHD had resolved by 8 months (patient 1) and 3 yr (patient 2) of glucocorticoid replacement. GH replacement was therefore stopped, hypoglycemia did not recur, and over the subsequent 10 yr, growth and puberty have proceeded normally.

Conclusions:

1) Physiological glucocorticoid levels appear to be required for the development and function of the somatotrophs during infancy. 2) Eucortisolism, like euthyroidism, is required for the proper evaluation of GH secretory capacity. 3) The metabolic effect of GH replacement is essential for the maintenance of normoglycemia, especially in infants. And 4) targeted molecular investigations are a powerful tool to clarify the diagnosis in severely ill infants and to reevaluate the specific treatment they need.
Variants in STAT5B Associate with Serum TC and LDL-C Levels
Authors: Kornfeld Jan-Wilhelm , Isaacs Aaron , Vitart Veronique , Pospisilik J. Andrew , Meitinger Thomas , Gyllensten Ulf , Wilson James F. , Rudan Igor , Campbell Harry , Penninger Josef M. , Sexl Veronika , Moriggl Richard , van Duijn Cornelia , Pramstaller Peter P. , Hicks Andrew A.
Published: July 2011

Context:

Known genetic variants influencing serum lipid levels do not adequately account for the observed population variability of these phenotypes. The GH/signal transducers and activators of transcription (STAT) signaling pathway is an evolutionary conserved system that exerts strong effects on metabolism, including that of lipids.

Research Design and Methods:

We analyzed the association of 11 single-nucleotide polymorphisms (SNP) spanning the STAT5B/STAT5A/STAT3 locus with serum lipid levels in six European populations (n = 5162 nondiabetic individuals).

Results:

After adjustment for age, sex, alcohol use, smoking, and body mass index, we identified STAT5B variants (rs8082391 and rs8064638) in novel association with total cholesterol (TC; P = 0.001 and P = 0.002) and low-density lipoprotein cholesterol (P = 0.002 and P = 0.004) levels. The minor alleles of these single-nucleotide polymorphisms were significantly enriched in hyperlipidemic individuals across the six discovery populations (P = 0.004 and P = 0.006). In transgenic mice deficient for hepatic STAT5A and STAT5B, reduced serum TC levels coincided with reduced hepatic cholesterol biosynthesis as demonstrated using gene expression profiling and pathway enrichment analysis.

Conclusions:

Genetic variants in STAT5B are associated with TC and low-density lipoprotein cholesterol levels among six populations. Mechanistically, STAT5B transcriptionally regulates hepatic cholesterol homeostasis.
Central Diabetes Insipidus as the Inaugural Manifestation of Langerhans Cell Histiocytosis: Natural History and Medical Evaluation of 26 Children and Adolescents
Authors: Marchand Isis , Barkaoui Mohamed Aziz , Garel Catherine , Polak Michel , Donadieu Jean ,   for the Writing Committee
Published: July 2011

Context:

Isolated central diabetes insipidus (CDI) can be the first manifestation of Langerhans cell histiocytosis (LCH), creating diagnostic dilemmas such as dysgerminoma and other inflammatory lesions.

Method:

In 2010, the French national LCH registry had enrolled 1236 LCH patients under 18 yr of age. Isolated CDI was the initial presentation of LCH in 26 patients. We reviewed their clinical and magnetic resonance imaging (MRI) features.

Results:

Median age at the diagnosis of CDI was 9.6 yr (1.8–16.3), and median follow-up after CDI diagnosis was 9.9 yr (3.5–26.6). In addition to CDI, two patients had visual field defects, four had secondary amenorrhea, and 11 had anterior pituitary deficiency. Cerebral imaging (including computed tomography in two cases), performed in 22 patients within 3 months of CDI diagnosis, showed pituitary stalk thickening in 14 patients, which was moderate (3.0–7 mm) in nine cases and marked (>7 mm) in five cases. In eight cases, the lesion extended to the floor of the third ventricle. One child with LCH presented with a mild enlarged sellar content. During follow-up, 22 patients developed extrapituitary involvement, mainly of bone (n = 15), lung (n = 9), and skin (n = 9). Pituitary biopsy was performed in eight cases and was conclusive in six cases.

Conclusions:

Pituitary stalk thickening can be observed in LCH as well as lesions extending to the floor of the third ventricle. In all cases but one, the intrasellar content was not enlarged. Long-term follow-up with close attention to bone, skin, and lung disorders may lead to the diagnosis of LCH.
SDHA Immunohistochemistry Detects Germline SDHA Gene Mutations in Apparently Sporadic Paragangliomas and Pheochromocytomas
Authors: Korpershoek Esther , Favier Judith , Gaal José , Burnichon Nelly , van Gessel Bram , Oudijk Lindsey , Badoual Cécile , Gadessaud Noémie , Venisse Annabelle , Bayley Jean-Pierre , van Dooren Marieke F. , de Herder Wouter W. , Tissier Frédérique , Plouin Pierre-François , van Nederveen Francien H. , Dinjens Winand N. M. , Gimenez-Roqueplo Anne-Paule , de Krijger Ronald R.
Published: July 2011

Context:

Pheochromocytoma-paraganglioma syndrome is caused by mutations in SDHB, SDHC, and SDHD, encoding subunits of succinate dehydrogenase (SDH), and in SDHAF2, required for flavination of SDHA. A recent report described a patient with an abdominal paraganglioma, immunohistochemically negative for SDHA, and identified a causal germline mutation in SDHA.

Objective:

In this study, we evaluated the significance of SDHA immunohistochemistry in the identification of new patients with SDHA mutations.

Setting:

This study was performed in the Erasmus Medical Center in Rotterdam (The Netherlands) and the Université Paris Descartes in Paris (France).

Methods:

We investigated 316 pheochromocytomas and paragangliomas for SDHA expression. Sequence analysis of SDHA was performed on all tumors that were immunohistochemically negative for SDHA and on a subset of tumors immunohistochemically positive for SDHA.

Results:

Six tumors were immunohistochemically negative for SDHA. Four tumors from Dutch patients showed a germline c.91C→T SDHA gene mutation (p.Arg31X). Another tumor (from France) carried a germline SDHA missense mutation c.1753C→T (p.Arg585Trp). Loss of the wild-type SDHA allele was confirmed by loss of heterozygosity analysis. Sequence analysis of 35 SDHA immunohistochemically positive tumors did not reveal additional SDHA mutations.

Conclusions:

Our results demonstrate that SDHA immunohistochemistry on paraffin-embedded tumors can reveal the presence of SDHA germline mutations and allowed the identification of SDHA-related tumors in at least 3% of patients affected by apparently sporadic (para)sympathetic paragangliomas and pheochromocytomas.
Follicle-Stimulating Hormone Receptor Polymorphism (G−29A) Is Associated with Altered Level of Receptor Expression in Granulosa Cells
Authors: Desai Swapna S. , Achrekar Swati K. , Pathak Bhakti R. , Desai Sadhana K. , Mangoli Vijay S. , Mangoli Ranjana V. , Mahale Smita D.
Published: July 2011

Context:

Polymorphisms of the FSHR gene are associated with variable ovarian response to FSH stimulation in subjects undergoing in vitro fertilization (IVF) treatment. The type of ovarian response is correlated with the level of FSH receptor (FSHR) expression on granulosa cells.

Objective:

We investigated whether the polymorphism at position −29 in the promoter of the FSHR gene may contribute in altered receptor expression.

Design and patients:

FSHR polymorphism at position −29 was studied in 100 subjects undergoing IVF treatment. Association of this polymorphism with level of FSHR expression was retrospectively analyzed.

Setting:

The study was conducted at an academic research institute and private IVF clinic.

Methods:

The genotype at position −29 of the FSHR gene was studied in IVF subjects by PCR-restriction fragment length polymorphism. Total RNA and protein was extracted from granulosa cells. The relative FSHR mRNA expression was carried out by real-time PCR. The receptor protein expression was evaluated by Western blot and confocal microscopy.

Results:

The clinical and endocrinological parameters revealed that almost 72% of subjects with the AA genotype at position −29 of FSHR gene were poor ovarian responders (odds ratio 8.63, 95% confidential interval 1.84–45.79; P = 0.001). The lower cleavage intensity predicted by in silico analysis for A allele as compared with the G allele suggest the difference in the DNA-protein binding affinity. The relative expression of FSHR at mRNA and protein level was significantly reduced in subjects with AA genotype as compared with the GG genotype.

Conclusion:

Poor ovarian response observed in subjects with the AA genotype at position −29 of the FSHR gene is due to reduced receptor expression.
Female gametophytic cell specification and seed development require the function of the putativeArabidopsisINCENP orthologWYRD
Authors: Kirioukhova Olga , Johnston Amal J. , Kleen Daniela , Kägi Christina , Baskar Ramamurthy , Moore James M. , Bäumlein Helmut , Groß-Hardt Rita , Grossniklaus Ueli
Published: July 2011

In plants, gametes, along with accessory cells, are formed by the haploid gametophytes through a series of mitotic divisions, cell specification and differentiation events. How the cells in the female gametophyte of flowering plants differentiate into gametes (the egg and central cell) and accessory cells remains largely unknown. In a screen for mutations that affect egg cell differentiation in Arabidopsis, we identified the wyrd (wyr) mutant, which produces additional egg cells at the expense of the accessory synergids. WYR not only restricts gametic fate in the egg apparatus, but is also necessary for central cell differentiation. In addition, wyr mutants impair mitotic divisions in the male gametophyte and endosperm, and have a parental effect on embryo cytokinesis, consistent with a function of WYR in cell cycle regulation. WYR is upregulated in gametic cells and encodes a putative plant ortholog of the inner centromere protein (INCENP), which is implicated in the control of chromosome segregation and cytokinesis in yeast and animals. Our data reveal a novel developmental function of the conserved cell cycle-associated INCENP protein in plant reproduction, in particular in the regulation of egg and central cell fate and differentiation.
A late phase of germ plasm accumulation during Drosophila oogenesis requires Lost and Rumpelstiltskin
Authors: Sinsimer Kristina S. , Jain Roshan A. , Chatterjee Seema , Gavis Elizabeth R.
Published: July 2011

Asymmetric mRNA localization is an effective mechanism for establishing cellular and developmental polarity. Posterior localization of oskar in the Drosophila oocyte targets the synthesis of Oskar to the posterior, where Oskar initiates the assembly of the germ plasm. In addition to harboring germline determinants, the germ plasm is required for localization and translation of the abdominal determinant nanos. Consequently, failure of oskar localization during oogenesis results in embryos lacking germ cells and abdominal segments. oskar accumulates at the oocyte posterior during mid-oogenesis through a well-studied process involving kinesin-mediated transport. Through live imaging of oskar mRNA, we have uncovered a second, mechanistically distinct phase of oskar localization that occurs during late oogenesis and results in amplification of the germ plasm. Analysis of two newly identified oskar localization factors, Rumpelstiltskin and Lost, that are required specifically for this late phase of oskar localization shows that germ plasm amplification ensures robust abdomen and germ cell formation during embryogenesis. In addition, our results indicate the importance of mechanisms for adapting mRNAs to utilize multiple localization pathways as necessitated by the dramatic changes in ovarian physiology that occur during oogenesis.
Specific and integrated roles of Lmx1a, Lmx1b and Phox2a in ventral midbrain development
Authors: Deng Qiaolin , Andersson Elisabet , Hedlund Eva , Alekseenko Zhanna , Coppola Eva , Panman Lia , Millonig James H. , Brunet Jean-Francois , Ericson Johan , Perlmann Thomas
Published: July 2011

The severe disorders associated with a loss or dysfunction of midbrain dopamine neurons (DNs) have intensified research aimed at deciphering developmental programs controlling midbrain development. The homeodomain proteins Lmx1a and Lmx1b are important for the specification of DNs during embryogenesis, but it is unclear to what degree they may mediate redundant or specific functions. Here, we provide evidence showing that DN progenitors in the ventral midbrain can be subdivided into molecularly distinct medial and lateral domains, and these subgroups show different sensitivity to the loss of Lmx1a and Lmx1b. Lmx1a is specifically required for converting non-neuronal floor-plate cells into neuronal DN progenitors, a process that involves the establishment of Notch signaling in ventral midline cells. On the other hand, lateral DN progenitors that do not appear to originate from the floor plate are selectively ablated in Lmx1b mutants. In addition, we also reveal an unanticipated role for Lmx1b in regulating Phox2a expression and the sequential specification of ocular motor neurons (OMNs) and red nucleus neurons (RNNs) from progenitors located lateral to DNs in the midbrain. Our data therefore establish that Lmx1b influences the differentiation of multiple neuronal subtypes in the ventral midbrain, whereas Lmx1a appears to be exclusively devoted to the differentiation of the DN lineage.
Microtubule-induced nuclear envelope fluctuations control chromatin dynamics in Drosophila embryos
Authors: Hampoelz Bernhard , Azou-Gros Yannick , Fabre Roxane , Markova Olga , Puech Pierre-Henri , Lecuit Thomas
Published: July 2011

Nuclear shape is different in stem cells and differentiated cells and reflects important changes in the mechanics of the nuclear envelope (NE). The current framework emphasizes the key role of the nuclear lamina in nuclear mechanics and its alterations in disease. Whether active stress controls nuclear deformations and how this stress interplays with properties of the NE to control NE dynamics is unclear. We address this in the early Drosophila embryo, in which profound changes in NE shape parallel the transcriptional activation of the zygotic genome. We show that microtubule (MT) polymerization events produce the elementary forces necessary for NE dynamics. Moreover, large-scale NE deformations associated with groove formation require concentration of MT polymerization in bundles organized by Dynein. However, MT bundles cannot produce grooves when the farnesylated inner nuclear membrane protein Kugelkern (Kuk) is absent. Although it increases stiffness of the NE, Kuk also stabilizes NE deformations emerging from the collective effect of MT polymerization forces concentrated in bundles. Finally, we report that MT-induced NE deformations control the dynamics of chromatin and its organization at steady state. Thus, the NE is a dynamic organelle, fluctuations of which increase chromatin dynamics. We propose that such mechanical regulation of chromatin dynamics by MTs might be important for gene regulation.