bluuebunny/crossref_metadata_2025 huggingface dataset:
An S/T-Q cluster domain census unveils new putative targets under Tel1/Mec1 control
Authors: Cheung Hannah C , San Lucas F Anthony , Hicks Stephanie , Chang Kyle , Bertuch Alison A , Ribes-Zamora Albert
Published: November 2012

Background

The cellular response to DNA damage is immediate and highly coordinated in order to maintain genome integrity and proper cell division. During the DNA damage response (DDR), the sensor kinases Tel1 and Mec1 in Saccharomyces cerevisiae and ATM and ATR in human, phosphorylate multiple mediators which activate effector proteins to initiate cell cycle checkpoints and DNA repair. A subset of kinase substrates are recognized by the S/T-Q cluster domain (SCD), which contains motifs of serine (S) or threonine (T) followed by a glutamine (Q). However, the full repertoire of proteins and pathways controlled by Tel1 and Mec1 is unknown.

Results

To identify all putative SCD-containing proteins, we analyzed the distribution of S/T-Q motifs within verified Tel1/Mec1 targets and arrived at a unifying SCD definition of at least 3 S/T-Q within a stretch of 50 residues. This new SCD definition was used in a custom bioinformatics pipeline to generate a census of SCD-containing proteins in both yeast and human. In yeast, 436 proteins were identified, a significantly larger number of hits than were expected by chance. These SCD-containing proteins did not distribute equally across GO-ontology terms, but were significantly enriched for those involved in processes related to the DDR. We also found a significant enrichment of proteins involved in telophase and cytokinesis, protein transport and endocytosis suggesting possible novel Tel1/Mec1 targets in these pathways. In the human proteome, a wide range of similar proteins were identified, including homologs of some SCD-containing proteins found in yeast. This list also included high concentrations of proteins in the Mediator, spindle pole body/centrosome and actin cytoskeleton complexes.

Conclusions

Using a bioinformatic approach, we have generated a census of SCD-containing proteins that are involved not only in known DDR pathways but several other pathways under Tel1/Mec1 control suggesting new putative targets for these kinases.
Assessment of virulence diversity of methicillin-resistant Staphylococcus aureus strains with a Drosophila melanogaster infection model
Authors: Wu Kaiyu , Conly John , Surette Michael , Sibley Christopher , Elsayed Sameer , Zhang Kunyan
Published: November 2012

Background

Staphylococcus aureus strains with distinct genetic backgrounds have shown different virulence in animal models as well as associations with different clinical outcomes, such as causing infection in the hospital or the community. With S. aureus strains carrying diverse genetic backgrounds that have been demonstrated by gene typing and genomic sequences, it is difficult to compare these strains using mammalian models. Invertebrate host models provide a useful alternative approach for studying bacterial pathogenesis in mammals since they have conserved innate immune systems of biological defense. Here, we employed Drosophila melanogaster as a host model for studying the virulence of S. aureus strains.

Results

Community-associated methicillin-resistant S. aureus (CA-MRSA) strains USA300, USA400 and CMRSA2 were more virulent than a hospital-associated (HA)-MRSA strain (CMRSA6) and a colonization strain (M92) in the D. melanogaster model. These results correlate with bacterial virulence in the Caenorhabditis elegans host model as well as human clinical data. Moreover, MRSA killing activities in the D. melanogaster model are associated with bacterial replication within the flies. Different MRSA strains induced similar host responses in D. melanogaster, but demonstrated differential expression of common bacterial virulence factors, which may account for the different killing activities in the model. In addition, hemolysin α, an important virulence factor produced by S. aureus in human infections is postulated to play a role in the fly killing.

Conclusions

Our results demonstrate that the D. melanogaster model is potentially useful for studying S. aureus pathogenicity. Different MRSA strains demonstrated diverse virulence in the D. melanogaster model, which may be the result of differing expression of bacterial virulence factors in vivo.
Highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of Listeria monocytogenes and L. ivanovii
Authors: Mendonça Marcelo , Conrad Neida L , Conceição Fabricio R , Moreira Ângela N , da Silva Wladimir P , Aleixo José AG , Bhunia Arun K
Published: November 2012

Background

Immunomagnetic separation (IMS) and immunoassays are widely used for pathogen detection. However, novel technology platforms with highly selective antibodies are essential to improve detection sensitivity, specificity and performance. In this study, monoclonal antibodies (MAbs) against Internalin A (InlA) and p30 were generated and used on paramagnetic beads of varying diameters for concentration, as well as on fiber-optic sensor for detection.

Results

Anti-InlA MAb-2D12 (IgG2a subclass) was specific for Listeria monocytogenes and L. ivanovii, and p30-specific MAb-3F8 (IgM) was specific for the genus Listeria. At all bacterial concentrations (103–108 CFU/mL) tested in the IMS assay; the 1-μm diameter MyOne beads had significantly higher capture efficiency (P < 0.05) than the 2.8-μm diameter M-280 beads with both antibodies. The highest capture efficiency for MyOne-2D12 (49.2% for 105 CFU/mL) was significantly higher (P < 0.05) than that of MyOne-3F8 (16.6 %) and Dynabeads anti-Listeria antibody (9 %). Furthermore, capture efficiency for MyOne-2D12 was highly specific for L. monocytogenes and L. ivanovii. Subsequently, we captured L. monocytogenes by MyOne-2D12 and MyOne-3F8 from hotdogs inoculated with mono- or co-cultures of L. monocytogenes and L. innocua (10–40 CFU/g), enriched for 18 h and detected by fiber-optic sensor and confirmed by plating, light-scattering, and qPCR assays. The detection limit for L. monocytogenes and L. ivanovii by the fiber-optic immunosensor was 3 × 102 CFU/mL using MAb-2D12 as capture and reporter antibody. Selective media plating, light-scattering, and qPCR assays confirmed the IMS and fiber-optic results.

Conclusions

IMS coupled with a fiber-optic sensor using anti-InlA MAb is highly specific for L. monocytogenes and L. ivanovii and enabled detection of these pathogens at low levels from buffer or food.
MicroRNA-99a induces G1-phase cell cycle arrest and suppresses tumorigenicity in renal cell carcinoma
Authors: Cui Li , Zhou Hua , Zhao Hu , Zhou Yaojun , Xu Renfang , Xu Xianlin , Zheng Lu , Xue Zhong , Xia Wei , Zhang Bo , Ding Tao , Cao Yunjie , Tian Zinong , Shi Qianqian , He Xiaozhou
Published: November 2012

Background

A growing body of evidence suggests that microRNAs (miRNAs) play an important role in cancer diagnosis and therapy. MicroRNA-99a (miR-99a), a potential tumor suppressor, is downregulated in several human malignancies. The expression and function of miR-99a, however, have not been investigated in human renal cell carcinoma (RCC) so far. We therefore examined the expression of miR-99a in RCC cell lines and tissues, and assessed the impact of miR-99a on the tumorigenesis of RCC.

Methods

MiR-99a levels in 40 pairs of RCC and matched adjacent non-tumor tissues were assessed by real-time quantitative Reverse Transcription PCR (qRT-PCR). The RCC cell lines 786-O and OS-RC-2 were transfected with miR-99a mimics to restore the expression of miR-99a. The effects of miR-99a were then assessed by cell proliferation, cell cycle, transwell, and colony formation assay. A murine xenograft model of RCC was used to confirm the effect of miR-99a on tumorigenicity in vivo. Potential target genes were identified by western blotting and luciferase reporter assay.

Results

We found that miR-99a was remarkably downregulated in RCC and low expression level of miR-99a was correlated with poor survival of RCC patients. Restoration of miR-99a dramatically suppressed RCC cells growth, clonability, migration and invasion as well as induced G1-phase cell cycle arrest in vitro. Moreover, intratumoral delivery of miR-99a could inhibit tumor growth in murine xenograft models of human RCC. In addition, we also fond that mammalian target of rapamycin (mTOR) was a direct target of miR-99a in RCC cells. Furthermore, siRNA-mediated knockdown of mTOR partially phenocopied the effect of miR-99a overexpression, suggesting that the tumor suppressive role of miR-99a may be mediated primarily through mTOR regulation.

Conclusions

Collectively, these results demonstrate for the first time, to our knowledge, that deregulation of miR-99a is involved in the etiology of RCC partially via direct targeting mTOR pathway, which suggests that miR-99a may offer an attractive new target for diagnostic and therapeutic intervention in RCC.
Screening of Finnish RAD51Cfounder mutations in prostate and colorectal cancer patients
Authors: Pelttari Liisa M , Nurminen Riikka , Gylfe Alexandra , Aaltonen Lauri A , Schleutker Johanna , Nevanlinna Heli
Published: November 2012

Background

Rare, heterozygous germline mutations in the RAD51C gene have been found in breast and ovarian cancer families. In the Finnish population, we have identified two founder mutations in RAD51C that increase the risk of ovarian cancer but not breast cancer in the absence of ovarian cancer. Risk for other cancers has not been studied.

Methods

To study the role of RAD51C mutations in other common cancer types, we genotyped the Finnish RAD51C founder mutations c.837 + 1G > A and c.93delG in 1083 prostate cancer patients and 802 colorectal cancer patients using TaqMan Real-Time PCR.

Results

No RAD51C mutations c.837 + 1G > A or c.93delG were detected among the prostate or colorectal cancer patients.

Conclusions

The results suggest that the RAD51C mutations do not predispose to prostate or colorectal cancer.
The LIFE child study: a life course approach to disease and health
Authors: Quante Mirja , Hesse Mara , Döhnert Mirko , Fuchs Michael , Hirsch Christian , Sergeyev Elena , Casprzig Nora , Geserick Mandy , Naumann Stephanie , Koch Christiane , Sabin Matthew A , Hiemisch Andreas , Körner Antje , Kiess Wieland ,   for the LIFE Child Study Investigators
Published: November 2012

Background

Profound knowledge about child growth, development, health, and disease in contemporary children and adolescents is still rare. Epidemiological studies together with new powerful research technologies present exciting opportunities to the elucidation of risk factor-outcome associations with potentially major consequences for prevention, diagnosis and treatment.

Aim

To conduct a unique prospective longitudinal cohort study in order to assess how environmental, metabolic and genetic factors affect growth, development and health from fetal life to adulthood.

Methods

The ‘Leipzig Research Centre for Civilization Diseases (LIFE) Child Study’ focuses on two main research objectives: (1) monitoring of normal growth, development and health; (2) non-communicable diseases such as childhood obesity and its co-morbidities, atopy and mental health problems. Detailed assessments will be conducted alongside long-term storage of biological samples in 2,000 pregnant women and more than 10,000 children and their families.

Results

Close coordination and engagement of a multidisciplinary team in the LIFE Child study successfully established procedures and systems for balancing many competing study and ethical needs. Full participant recruitment and complete data collection started in July 2011. Early data indicate a high acceptance rate of the study program, successful recruitment strategies and the establishment of a representative cohort for the population of Leipzig. A series of subprojects are ongoing, and analyses and publications are on their way.

Discussion

This paper addresses key elements in the design and implementation of the new prospective longitudinal cohort study LIFE Child. Given the recognized need for long-term data on adverse effects on health and protective factors, our study data collection should provide magnificent opportunities to examine complex interactions that govern the emergence of non-communicable diseases.
Wound repair and anti-inflammatory potential of Lonicera japonica in excision wound-induced rats
Authors: Chen Wei-Cheng , Liou Shorong-Shii , Tzeng Thing-Fong , Lee Shiow-Ling , Liu I-Min
Published: November 2012

Background

Lonicera japonica Thunb. (Caprifoliaceae), a widely used traditional Chinese medicinal plant, is used to treat some infectious diseases and it may have uses as a healthy food and applications in cosmetics and as an ornamental groundcover. The ethanol extract of the flowering aerial parts of L. japonica (LJEE) was investigated for its healing efficiency in a rat excision wound model.

Methods

Excision wounds were inflicted upon three groups of eight rats each. Healing was assessed by the rate of wound contraction in skin wound sites in rats treated with simple ointment base, 10% (w/w) LJEE ointment, or the reference standard drug, 0.2% (w/w) nitrofurazone ointment. The effects of LJEE on the contents of hydroxyproline and hexosamine during healing were estimated. The antimicrobial activity of LJEE against microorganisms was also assessed. The in vivo anti-inflammatory activity of LJEE was investigated to understand the mechanism of wound healing.

Results

LJEE exhibited significant antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Candida albicans, and Candida tropicalis. The ointment formulation prepared with 10% (w/w) LJEE exhibited potent wound healing capacity as evidenced by the wound contraction in the excision wound model. The contents of hydroxyproline and hexosamine also correlated with the observed healing pattern. These findings were supported by the histopathological characteristics of healed wound sections, as greater tissue regeneration, more fibroblasts, and angiogenesis were observed in the 10% (w/w) LJEE ointment-treated group. The results also indicated that LJEE possesses potent anti-inflammatory activity, as it enhanced the production of anti-inflammatory cytokines that suppress proinflammatory cytokine production.

Conclusions

The results suggest that the antimicrobial and anti-inflammatory activities of LJEE act synergistically to accelerate wound repair.
The effects of Sutaehwan-Gami on menopausal symptoms induced by ovariectomy in rats
Authors: Kim Dong-Il , Choi Min Sun , Pak Sok Cheon , Lee Seung-bok , Jeon Songhee
Published: November 2012

Background

This study was undertaken to evaluate the beneficial effects of a modified prescription of Sutaehwan named Sutaehwan-Gami (SG), created by adding Rhizoma dioscoreae and Carthami semen to Sutaehwan, on menopausal symptoms.

Methods

To evaluate the estrogenic effect of SG, we first examined estrogen receptor (ER) activation by SG treatment in breast adenocarcinoma cells and confirmed the estrogenic effect of SG in vivo ovariectomized rats. The animals were randomized into four groups: Sham operated group (Sham), saline treated ovariectomized group (OVX), SG treated group (SG) and raloxifene treated group (RLX). Animals were provided with SG at a dose of 500 mg/kg bw/day and RLX at a dose of 5.4 mg/kg bw/day with standard rat pellets for 3 months.

Results

SG significantly increased ERα phosphorylation, and its downstream effectors, extracellular signal-regulated kinase (ERK) and protein kinase B (Akt) phosphorylation in breast adenocarcinoma cells. Treatment with SG reversed ovariectomy-induced uterine weight reduction and weight gain. Decreases in the levels of GOT and GPT were observed in the SG group. The significantly reduced E2β level in OVX rats was raised by treatment with SG. Moreover, SG significantly increased the phosphorylation levels of ERK and Akt in the uterus.

Conclusion

Taken together, these data indicate that SG has phytoestrogen-like properties through ERK and Akt activation, implying that it could be protective and beneficial for the management of menopausal symptoms.
An antibody microarray analysis of serum cytokines in neurodegenerative Parkinsonian syndromes
Authors: Mahlknecht Philipp , Stemberger Sylvia , Sprenger Fabienne , Rainer Johannes , Hametner Eva , Kirchmair Rudolf , Grabmer Christoph , Scherfler Christoph , Wenning Gregor K , Seppi Klaus , Poewe Werner , Reindl Markus
Published: November 2012

Background

Microarray technology may offer a new opportunity to gain insight into disease-specific global protein expression profiles. The present study was performed to apply a serum antibody microarray to screen for differentially regulated cytokines in Parkinson's disease (PD), multiple system atrophy (MSA), progressive supranuclear palsy (PSP) and corticobasal syndrome (CBS).

Results

Serum samples were obtained from patients with clinical diagnoses of PD (n = 117), MSA (n = 31) and PSP/CBS (n = 38) and 99 controls. Cytokine profiles of sera from patients and controls were analyzed with a semiquantitative human antibody array for 174 cytokines and the expression of 12 cytokines was found to be significantly altered. In a next step, results from the microarray experiment were individually validated by different immunoassays. Immunoassay validation confirmed a significant increase of median PDGF-BB levels in patients with PSP/CBS, MSA and PD and a decrease of median prolactin levels in PD. However, neither PDGF-BB nor prolactin were specific biomarkers to discriminate PSP/CBS, MSA, PD and controls.

Conclusions

In our unbiased cytokine array based screening approach and validation by a different immunoassay only two of 174 cytokines were significantly altered between patients and controls.
Cardiac Depression Scale: Mokken scaling in heart failure patients
Authors: Ski Chantal F , Thompson David R , Hare David L , Stewart Andrew G , Watson Roger
Published: November 2012

Background

There is a high prevalence of depression in patients with heart failure (HF) that is associated with worsening prognosis. The value of using a reliable and valid instrument to measure depression in this population is therefore essential. We validated the Cardiac Depression Scale (CDS) in heart failure patients using a model of ordinal unidimensional measurement known as Mokken scaling.

Findings

We administered in face-to-face interviews the CDS to 603 patients with HF. Data were analysed using Mokken scale analysis. Items of the CDS formed a statistically significant unidimensional Mokken scale of low strength (H<0.40) and high reliability (Rho>0.8).

Conclusions

The CDS has a hierarchy of items which can be interpreted in terms of the increasingly serious effects of depression occurring as a result of HF. Identifying an appropriate instrument to measure depression in patients with HF allows for early identification and better medical management.
NADPH oxidase 2-derived reactive oxygen species signal contributes to bradykinin-induced matrix metalloproteinase-9 expression and cell migration in brain astrocytes
Authors: Lin Chih-Chung , Hsieh Hsi-Lung , Shih Ruey-Horng , Chi Pei-Ling , Cheng Shin-Ei , Chen Jin-Chung , Yang Chuen-Mao
Published: November 2012

Background

Matrix metalloproteinase-9 (MMP-9) plays a crucial role in pathological processes of brain inflammation, injury, and neurodegeneration. Moreover, bradykinin (BK) induces the expression of several inflammatory proteins in brain astrocytes. Recent studies have suggested that increased oxidative stress is implicated in the brain inflammation and injury. However, whether BK induced MMP-9 expression mediated through oxidative stress remains virtually unknown. Herein we investigated the role of redox signals in BK-induced MMP-9 expression in rat brain astrocytes (RBA-1 cells).

Results

In the study, we first demonstrated that reactive oxygen species (ROS) plays a crucial role in BK-induced MMP-9 expression in cultured brain astrocytes (in vitro) and animal brain tissue (in vivo) models. Next, BK-induced MMP-9 expression is mediated through a Ca2+-mediated PKC-α linking to p47phox/NADPH oxidase 2 (Nox2)/ROS signaling pathway. Nox2-dependent ROS generation led to activation and up-regulation of the downstream transcriptional factor AP-1 (i.e. c-Fos and c-Jun), which bound to MMP-9 promoter region, and thereby turned on transcription of MMP-9 gene. Functionally, BK-induced MMP-9 expression enhanced astrocytic migration.

Conclusions

These results demonstrated that in RBA-1 cells, activation of AP-1 (c-Fos/c-Jun) by the PKC-α-mediated Nox2/ROS signals is essential for up-regulation of MMP-9 and cell migration enhanced by BK.
Transcriptomics and proteomics analyses of the PACAP38 influenced ischemic brain in permanent middle cerebral artery occlusion model mice
Authors: Hori Motohide , Nakamachi Tomoya , Rakwal Randeep , Shibato Junko , Ogawa Tetsuo , Aiuchi Toshihiro , Tsuruyama Tatsuaki , Tamaki Keiji , Shioda Seiji
Published: November 2012

Introduction

The neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) is considered to be a potential therapeutic agent for prevention of cerebral ischemia. Ischemia is a most common cause of death after heart attack and cancer causing major negative social and economic consequences. This study was designed to investigate the effect of PACAP38 injection intracerebroventrically in a mouse model of permanent middle cerebral artery occlusion (PMCAO) along with corresponding SHAM control that used 0.9% saline injection.

Methods

Ischemic and non-ischemic brain tissues were sampled at 6 and 24 hours post-treatment. Following behavioral analyses to confirm whether the ischemia has occurred, we investigated the genome-wide changes in gene and protein expression using DNA microarray chip (4x44K, Agilent) and two-dimensional gel electrophoresis (2-DGE) coupled with matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS), respectively. Western blotting and immunofluorescent staining were also used to further examine the identified protein factor.

Results

Our results revealed numerous changes in the transcriptome of ischemic hemisphere (ipsilateral) treated with PACAP38 compared to the saline-injected SHAM control hemisphere (contralateral). Previously known (such as the interleukin family) and novel (Gabra6, Crtam) genes were identified under PACAP influence. In parallel, 2-DGE analysis revealed a highly expressed protein spot in the ischemic hemisphere that was identified as dihydropyrimidinase-related protein 2 (DPYL2). The DPYL2, also known as Crmp2, is a marker for the axonal growth and nerve development. Interestingly, PACAP treatment slightly increased its abundance (by 2-DGE and immunostaining) at 6 h but not at 24 h in the ischemic hemisphere, suggesting PACAP activates neuronal defense mechanism early on.

Conclusions

This study provides a detailed inventory of PACAP influenced gene expressions and protein targets in mice ischemic brain, and suggests new targets for thereaupetic interventions.
Induction of cross-protection against influenza A virus by DNA prime-intranasal protein boost strategy based on nucleoprotein
Authors: Luo Jian , Zheng Dan , Zhang Wenjie , Fang Fang , Wang Hanzhong , Sun Ying , Ding Yahong , Xu Chengfei , Chen Quanjiao , Zhang Hongbo , Huang Ding , Sun Bing , Chen Ze
Published: November 2012

Background

The highly conserved nucleoprotein (NP) is an internal protein of influenza virus and is capable of inducing cross-protective immunity against different influenza A viruses, making it a main target of universal influenza vaccine. In current study, we characterized the immune response induced by DNA prime-intranasal protein boost strategy based on NP (A/PR/8/34, H1N1) in mouse model, and evaluated its protection ability against a lethal dose challenge of influenza virus.

Results

The intranasal boost with recombinant NP (rNP) protein could effectively enhance the pre-immune response induced by the NP DNA vaccine in mice. Compared to the vaccination with NP DNA or rNP protein alone, the prime-boost strategy increased the level of NP specific serum antibody, enhanced the T cell immune response, and relatively induced more mucosal IgA antibody. The overall immune response induced by this heterologous prime-boost regimen was Th-1-biased. Furthermore, the immune response in mice induced by this strategy provided not only protection against the homologous virus but also cross-protection against a heterosubtypic H9N2 strain.

Conclusions

The NP DNA prime-intranasal protein boost strategy may provide an effective strategy for universal influenza vaccine development.
p300 Exerts an Epigenetic Role in Chronic Neuropathic Pain through its Acetyltransferase Activity in Rats following Chronic Constriction Injury (CCI)
Authors: Zhu Xiao-Yan , Huang Chang-Sheng , Li Qian , Chang Rui-Min , Song Zong-Bing , Zou Wang-Yuan , Guo Qu-Lian
Published: November 2012

Background:

Neuropathic pain is detrimental to human health; however, its pathogenesis still remains largely unknown. Overexpression of pain-associated genes and increased nociceptive somato-sensitivity are well observed in neuropathic pain. The importance of epigenetic mechanisms in regulating the expression of pro- or anti-nociceptive genes has been revealed by studies recently, and we hypothesize that the transcriptional coactivator and the histone acetyltransferase E1A binding protein p300 (p300), as a part of the epigenetic mechanisms of gene regulation, may be involved in the pathogenesis of neuropathic pain induced by chronic constriction injury (CCI). To test this hypothesis, two different approaches were used in this study: (I) down-regulating p300 with specific small hairpin RNA (shRNA) and (II) chemical inhibition of p300 acetyltransferase activity by a small molecule inhibitor, C646.

Results:

Using the CCI rat model, we found that the p300 expression was increased in the lumbar spinal cord on day 14 after CCI. The treatment with intrathecal p300 shRNA reversed CCI-induced mechanical allodynia and thermal hyperalgesia, and suppressed the expression of cyclooxygenase-2 (COX-2), a neuropathic pain-associated factor. Furthermore, C646, an inhibitor of p300 acetyltransferase, also attenuated mechanical allodynia and thermal hyperalgesia, accompanied by a suppressed COX-2 expression, in the spinal cord.

Conclusions:

The results suggest that, through its acetyltransferase activity in the spinal cord after CCI, p300 epigenetically plays an important role in neuropathic pain. Inhibiting p300, using interfering RNA or C646, may be a promising approach to the development of new neuropathic pain therapies.
Facile spectrophotometric assay of molar equivalents of N-hydroxysuccinimide esters of monomethoxyl poly-(ethylene glycol) derivatives
Authors: Gao Ang , Yang Xiaolan , Zhang Chun , Long Gaobo , Pu Jun , Yuan Yonghua , Liu Hongbo , Li Yuanli , Liao Fei
Published: November 2012

Background

A new method is developed to quantify molar equivalents of N-hydroxysuccinimide (NHS) esters of derivatives of monomethoxyl poly-(ethylene glycol) (mPEG) in their preparations with NHS acetate ester as the reference.

Results

NHS ester of succinic monoester or carbonate of mPEG of 5,000 Da was synthesized and reacted with excessive ethanolamine in dimethylformamide at 25°C for 15 min. Residual ethanolamine was subsequently quantified by absorbance at 420 nm after reaction with 2,4,6-trinitrobenzenesulfonic acid (TNBS) at pH 9.2 for 15 min at 55°C followed by cooling with tap water. Reaction products of ethanolamine and NHS esters of mPEG caused no interference with TNBS assay of residual ethanolamine. Reaction between ethanolamine and NHS acetate ester follows 1:1 stoichiometry. By the new method, molar equivalents of NHS esters of carbonate and succinic monoester of mPEG in their preparations were about 90% and 60% of their theoretical values, respectively. During storage at 37°C in humid air, the new method detected spontaneous hydrolyses of the two NHS esters of mPEG more sensitively than the classical spectrophotometric method based on absorbance at 260 nm of NHS released by reaction with ammonia in aqueous solution.

Conclusion

The new method is favorable to quantify molar equivalents of NHS esters of mPEG derivatives and thus control quality of their preparations.
Perivascular mesenchymal stem cells in the adult human brain: a future target for neuroregeneration?
Authors: Özen Ilknur , Boix Jordi , Paul Gesine
Published: November 2012

Perivascular adult stem cells have been isolated from several tissues, including the adult human brain. They have unique signatures resembling both pericytes and mesenchymal stem cells. Understanding the nature of these cells in their specific vascular niches is important to determine their clinical potential as a new adult stem cell source. Indeed, they have promising features in vitro in terms of multipotency, immunomodulation and secretion of growth factors and cytokines. However, their in vivo function is less known as yet. Recent emerging data show a crucial role of perivascular mesenchymal stem cells in tissue homeostasis and repair. Furthermore, these cells may play an important role in adult stem cell niche regulation and in neurodegeneration. Here we review the recent literature on perivascular mesenchymal stem cells, discuss their different in vitro functions and highlight especially the specific properties of brain‐derived perivascular mesenchymal stem cells. We summarize current evidence that suggests an important in vivo function of these cells in terms of their regenerative potential that may indicate a new target cell for endogenous tissue regeneration and repair.
Adherence to a treat-to-target strategy in early rheumatoid arthritis: results of the DREAM remission induction cohort
Authors: Vermeer Marloes , Kuper Hillechiena H , Bernelot Moens Hein J , Hoekstra Monique , Posthumus Marcel D , van Riel Piet LCM , van de Laar Mart AFJ
Published: November 2012

Introduction

Clinical trials have demonstrated that treatment-to-target (T2T) is effective in achieving remission in early rheumatoid arthritis (RA). However, the concept of T2T has not been fully implemented yet and the question is whether a T2T strategy is feasible in daily clinical practice. The objective of the study was to evaluate the adherence to a T2T strategy aiming at remission (Disease Activity Score in 28 joints (DAS28) < 2.6) in early RA in daily practice. The recommendations regarding T2T included regular assessment of the DAS28 and advice regarding DAS28-driven treatment adjustments.

Methods

A medical chart review was performed among a random sample of 100 RA patients of the DREAM remission induction cohort. At all scheduled visits, it was determined whether the clinical decisions were compliant to the T2T recommendations.

Results

The 100 patients contributed to a total of 1,115 visits. The DAS28 was available in 97.9% (1,092/1,115) of the visits, of which the DAS28 was assessed at a frequency of at least every three months in 88.3% (964/1,092). Adherence to the treatment advice was observed in 69.3% (757/1,092) of the visits. In case of non-adherence when remission was present (19.5%, 108/553), most frequently medication was tapered off or discontinued when it should have been continued (7.2%, 40/553) or treatment was continued when it should have been tapered off or discontinued (6.2%, 34/553). In case of non-adherence when remission was absent (42.1%, 227/539), most frequently medication was not intensified when an intensification step should have been taken (34.9%, 188/539). The main reason for non-adherence was discordance between disease activity status according to the rheumatologist and DAS28.

Conclusions

The recommendations regarding T2T were successfully implemented and high adherence was observed. This demonstrates that a T2T strategy is feasible in RA in daily clinical practice.
Prevalence of Germline TP53 Mutations in a Prospective Series of Unselected Patients with Adrenocortical Carcinoma
Authors: Raymond Victoria M. , Else Tobias , Everett Jessica N. , Long Jessica M. , Gruber Stephen B. , Hammer Gary D.
Published: November 2012

Purpose:

Adrenocortical carcinoma (ACC) is a hallmark cancer in families with Li Fraumeni syndrome (LFS) caused by mutations in the TP53 gene. The prevalence of germline TP53 mutations in children diagnosed with ACC ranges from 50–97%. Although existing criteria advocate for TP53 testing in all patients with ACC regardless of age at diagnosis, the overall prevalence of germline mutations in patients diagnosed with ACC has not been well studied.

Patients and Methods:

A total of 114 patients with confirmed ACC evaluated in the University of Michigan Endocrine Oncology Clinic were prospectively offered genetic counseling and TP53 genetic testing, regardless of age at diagnosis or family history. Ninety-four of the 114 patients met with a genetic counselor (82.5%), with 53 of 94 (56.4%) completing TP53 testing; 9.6% (nine of 94) declined testing. The remainder (32 of 94; 34%) expressed interest in testing but did not pursue it for various reasons.

Results:

Four of 53 patients in this prospective, unselected series were found to have a TP53 mutation (7.5%). The prevalence of mutations in those diagnosed over age 18 was 5.8% (three of 52). There were insufficient data to estimate the prevalence in those diagnosed under age 18. None of these patients met clinical diagnostic criteria for classic LFS. Three of the families met criteria for Li Fraumeni-like syndrome; one patient met no existing clinical criteria for LFS or Li Fraumeni-like syndrome. Three of the four patients with mutations were diagnosed with ACC after age 45.

Conclusions:

Genetic counseling and germline testing for TP53 should be offered to all patients with ACC. Restriction on age at diagnosis or strength of the family history would fail to identify mutation carriers.
Visualization of cell cycle in mouse embryos with Fucci2 reporter directed by Rosa26 promoter
Authors: Abe Takaya , Sakaue-Sawano Asako , Kiyonari Hiroshi , Shioi Go , Inoue Ken-ichi , Horiuchi Toshitaka , Nakao Kazuki , Miyawaki Atsushi , Aizawa Shinichi , Fujimori Toshihiko
Published: November 2012

Fucci technology makes possible the distinction between live cells in the G1 and S/G2/M phases by dual-color imaging. This technology relies upon ubiquitylation-mediated proteolysis, and transgenic mice expressing Fucci provide a powerful model system with which to study the coordination of the cell cycle and development. The mice were initially generated using the CAG promoter; lines expressing the G1 and S/G2/M phase probes that emitted orange (mKO2) and green (mAG) fluorescence, respectively, were separately constructed. Owing to cell type-biased strength of the CAG promoter as well as the positional effects of random transgenesis, however, we noticed some variability in Fucci expression levels. To control more reliably the expression of cell cycle probes, we used different genetic approaches to create two types of reporter mouse lines with Fucci2 and Rosa26 transcriptional machinery. Fucci2 is a recently developed Fucci derivative, which emits red (mCherry) and green (mVenus) fluorescence and provides better color contrast than Fucci. A new transgenic line, R26p-Fucci2, utilizes the Rosa26 promoter and harbors the G1 and S/G2/M phase probes in a single transgene to preserve their co-inheritance. In the other R26R-Fucci2 approach, the two probes are incorporated into Rosa26 locus conditionally. The Cre-mediated loxP recombination technique thus allows researchers to design cell-type-specific Fucci2 expression. By performing time-lapse imaging experiments using R26p-Fucci2 and R26-Fucci2 in which R26R-Fucci2 had undergone germline loxP recombination, we demonstrated the great promise of these mouse reporters for studying cell cycle behavior in vivo.
A transiently expressed connexin is essential for anterior neural plate development in Ciona intestinalis
Authors: Hackley Christopher , Mulholland Erin , Kim Gil Jung , Newman-Smith Erin , Smith William C.
Published: November 2012

A forward genetic screen in the ascidian Ciona intestinalis identified a mutant line (frimousse) with a profound disruption in neural plate development. In embryos with the frimousse mutation, the anteriormost neural plate cells, which are products of an FGF induction at the blastula and gastrula stages, initially express neural plate-specific genes but fail to maintain the induced state and ultimately default to epidermis. The genetic lesion in the frimousse mutant lies within a connexin gene (cx-11) that is transiently expressed in the developing neural plate in a temporal window corresponding to the period of a-lineage neural induction. Using a genetically encoded calcium indicator we observed multiple calcium transients throughout the developing neural plate in wild-type embryos, but not in mutant embryos. A series of treatments at the gastrula and neurula stages that block the calcium transients, including gap junction inhibition and calcium depletion, were also found to disrupt the development of the anterior neural plate in a similar way to the frimousse mutation. The requirement for cx-11 for anterior neural fate points to a crucial role for intercellular communication via gap junctions, probably through mediation of Ca2+ transients, in Ciona intestinalis neural induction.
In toto live imaging of mouse morphogenesis and new insights into neural tube closure
Authors: Massarwa R’ada , Niswander Lee
Published: November 2012

In the field of developmental biology, live imaging is a powerful tool for studying, in real time, the dynamic behaviors of tissues and cells during organ formation. Mammals, which develop in utero, have presented a challenge for live imaging. Here, we offer a novel, prolonged and robust live imaging system for visualizing the development of a variety of embryonic tissues in the midgestation mouse embryo. We demonstrate the advantages of this imaging system by following the dynamics of neural tube closure during mouse embryogenesis and reveal extensive movements of the cranial neural tissue that are independent of neural fold zipping.
Hedgehog is required for CySC self-renewal but does not contribute to the GSC niche in the Drosophila testis
Authors: Amoyel Marc , Sanny Justina , Burel Michael , Bach Erika A.
Published: November 2012

The Drosophila testis harbors two types of stem cells: germ line stem cells (GSCs) and cyst stem cells (CySCs). Both stem cell types share a physical niche called the hub, located at the apical tip of the testis. The niche produces the JAK/STAT ligand Unpaired (Upd) and BMPs to maintain CySCs and GSCs, respectively. However, GSCs also require BMPs produced by CySCs, and as such CySCs are part of the niche for GSCs. Here we describe a role for another secreted ligand, Hedgehog (Hh), produced by niche cells, in the self-renewal of CySCs. Hh signaling cell-autonomously regulates CySC number and maintenance. The Hh and JAK/STAT pathways act independently and non-redundantly in CySC self-renewal. Finally, Hh signaling does not contribute to the niche function of CySCs, as Hh-sustained CySCs are unable to maintain GSCs in the absence of Stat92E. Therefore, the extended niche function of CySCs is solely attributable to JAK/STAT pathway function.
A continuum of transcriptional identities visualized by combinatorial fluorescent in situ hybridization
Authors: Jakt Lars Martin , Moriwaki Satoko , Nishikawa Shinichi
Published: November 2012

Oligonucleotide-based fluorescent in situ hybridization (FISH) coupled with high-resolution high-sensitivity microscopy allows the visualization of single RNA molecules within fixed cells and tissues as distinct foci. We show here that combinatorial labeling of RNA molecules with several fluorescent dyes extends the number of genes that can be targeted simultaneously beyond the number of fluorophores used. This approach also inherently validates the identification of transcripts reducing false positive counts. We have used combinatorial FISH and image analysis to measure the transcript densities of six genes using three fluorophores. This has allowed us to visualize the endothelial maturation of lateral mesoderm in an in vitro ES differentiation assay from a single snapshot of molecular identities. Our observations show that, under these specific conditions, endothelial maturation follows a homogeneous course with a gradual increase in expression of Cdh5 and a concomitant loss of early transcription factors, arguing that maturation is governed in a generally deterministic manner. This methodology is limited by the number of fluorophores that can be used and by the available microscopic resolution, but currently available equipment should allow the visualization of transcripts from 10 or more genes simultaneously.
Cu–Rh Redox Relay Catalysts for Synthesis of Azaheterocycles via C–H Functionalization
Authors: Chiba Shunsuke
Published: November 2012

This Highlight Review describes a Cu–Rh bimetallic redox relay catalytic system that enables efficient synthesis of highly substituted isoquinolines and their derivatives from readily available aryl ketoximes or α-arylvinyl azides with internal alkynes via C–H bond functionalization. A preliminary mechanistic investigation revealed that both the Cu and Rh catalysts are prerequisites to achieve the present process, and play their particular roles with synergistic cooperation during the multistep sequence.
Magnetic Orientation of Hexagonal Carbon Layers at High Temperatures
Authors: Sakaguchi Ayumi , Hamasaki Atom , Sadatou Toyonari , Nishihara Yoshitaka , Yamamoto Shou , Sekinuma Yuya , Ozeki Sumio
Published: November 2012

Coal tar pitch was treated at 773 K in a constructed furnace system used under a magnetic flux density (B) of up to 10 T. The (002) X-ray diffraction peak intensity of the treated pitches increased with increasing B to approach a saturation value over 6 T. The magnetic-field dependence of the intensity was fitted by the order parameter calculated by assuming the cooperative orientation of several thousand carbon crystallites. This suggests that the hexagonal carbon layers should be oriented in parallel to the magnetic fields.
Dipole Moments of Amino Acid Residues, Gly and Ala, in α-Helix: Quantum Chemical Building Blocks for Macrodipole Moment of α-Helical Polypeptide
Authors: Mieda Shunsuke , Aida Misako
Published: November 2012

We present the dipole moments of amino acid residues, Gly and Ala, as the building blocks of α-helix, which inherently take account of the hydrogen bonding in the main chain of polypeptide in the circumstance of α-helix. The dipole moments can be used to predict the macrodipole moments of longer polypeptides in α-helix.
In Vivo Real-time Detection of Plant Response to Physical and Chemical Stresses by Spin Probe ESR
Authors: Endo Mami , Kurosawa Hidehiro , Kawai Takahiro , Ito Tomohiro , Ogata Tateaki
Published: November 2012

It was possible to trace real-time change of redox balance in a plant in vivo by a spin probe-ESR method, after exposing it to various physical and chemical stresses. Consequently, a strong light, a cold spray, and NO2 and O3 gases were found to lead to relatively oxidative states in broccoli sprouts, but SO2 and CO2 did not influence the redox balance in the sprouts in the dark.
Mild Synthesis of Furans with a Quaternary Carbon Substituent at the 2-Position
Authors: Iwamoto Akihisa , Katori Aki , Sashihara Yoshiaki , Kojima Satoshi
Published: November 2012

The reaction of cis-4-oxopent-2-enal with β-carbonyl compounds in the presence of weak acids such as hexafluoroisopropyl alcohol was found to give furans with a quaternary carbon substituent at the 2-positon at room temperature or below. Reactivity was dependent on the identity of the substrates.
Phosphorescence Detection of the Singlet Molecular Oxygen Generated by Visible-light Irradiation on Gold-nanoparticle-deposited TiO2 Photocatalysts
Authors: Saito Hironobu , Nosaka Yoshio
Published: November 2012

We detected singlet molecular oxygen (1O2) with a near-infrared phosphorescence peak at 1270 nm in photocatalysis of gold-nanoparticle-deposited titanium dioxide (AuNP/TiO2) by the excitation of surface plasmon resonance. The formation of 1O2 indicates a possible application of AuNP/TiO2 for photodynamic therapy.
Mg2+ Storage in Organic Positive-electrode Active Material Based on 2,5-Dimethoxy-1,4-benzoquinone
Authors: Sano Hikaru , Senoh Hiroshi , Yao Masaru , Sakaebe Hikari , Kiyobayashi Tetsu
Published: November 2012

The potential capability of an organic positive-electrode active material for rechargeable magnesium batteries was investigated. 2,5-Dimethoxy-1,4-benzoquinone (DMBQ), which is known to work as a positive-electrode active material in the lithium system, was examined. The DMBQ electrode showed a discharge capacity of 260 mA h g(DMBQ)−1 and an average voltage of 0.9 V versus a magnesium quasi-reference electrode. The DMBQ electrode was charged and discharged reversibly with Mg2+ ions for several cycles.
Unexpected Extension of Usage of PPh3/CBr4, a Versatile Reagent: Isomerization of Aromatic Allylic Alcohols
Authors: Gong Wanchun , Liu Yun , Xue Jijun , Xie Zhixiang , Li Ying
Published: November 2012

The PPh3/CBr4-catalyzed isomerization of 2-aromatic allylic alcohols into the corresponding saturated aldehydes or ketones has been achieved at room temperature in good to excellent yields under mild and metal-free conditions. This new methodology has been applied successfully to the synthesis of ibuprofen in four steps.
A Simple Rhodamine-based Naked-eye and Fluorescence “Off–On” Sensor for Cu(II) in Aqueous Solution
Authors: Goswami Shyamaprosad , Manna Abhishek , Aich Krishnendu , Paul Sima
Published: November 2012

A new rhodamine-6G-based sensor is designed and synthesized. Its absorbance and fluorescence sensing behavior toward metal ions is investigated in mixed aqueous media (CH3OH/H2O, 6:4, v/v, pH 7.1). It displays selective Cu2+-induced spirolactam ring opening of the rhodamine dye in the presence of other heavy and transition-metal (HTM) ions.
Solid-State NMR Study on Actinide Dioxides
Authors: Tokunaga Yo , Sakai Hironori , Kambe Shinsaku , Chudo Hiroyuki , Osaka Masahiko , Miwa Shuhei , Nishi Tsuyoshi , Nakada Masami , Itoh Akinori , Homma Yoshiya
Published: November 2012

Besides the importance of the actinide dioxide series as a nuclear fuel, the magnetic properties of these compounds at low temperatures are particularly interesting. Their surprisingly varied physical properties at low temperatures stimulate continuing interest for both theory and experiment. Recently, we have performed 17O-NMR studies for the first time on Pu and Amcontaining dioxide systems, (Pu1-xAmx)O2. For the x=0.09 sample, a temperature-dependent NMR line broadening has been observed at low temperatures. By comparing the experimental data with the results of NMR line simulations, we have estimated the effective moment of Am ions to be Peff=1.38 μB. The value suggests the 5f5 (Am4+) state of the Am ion in PuO2. For the x=1 (=AmO2) sample, on the other hand, our 17O-NMR data provide the first microscopic evidence for a phase transition at 8.5 K as a bulk property in this system. A spectrum with a triangular line shape indicates that the internal field is distributed very nearly randomly in the ordered state.