bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumour stromal cells derived from paediatric malignancies display MSC-like properties and impair NK cell cytotoxicity
Authors: Johann Pascal-David , Vaegler Martin , Gieseke Friederike , Mang Philippa , Armeanu-Ebinger Sorin , Kluba Torsten , Handgretinger Rupert , Müller Ingo
Published: September 2010

Background

Tumour growth and metastatic infiltration are favoured by several components of the tumour microenvironment. Bone marrow-derived multipotent mesenchymal stromal cells (MSC) are known to contribute to the tumour stroma. When isolated from healthy bone marrow, MSC exert potent antiproliferative effects on immune effector cells. Due to phenotypic and morphological similarities of MSC and tumour stromal cells (TStrC), we speculated that immunotherapeutic approaches may be hampered if TStrC may still exhibit immunomodulatory properties of MSC.

Methods

In order to compare immunomodulatory properties of MSC and tumour stromal cells (TStrC), we established and analyzed TStrC cultures from eleven paediatric tumours and MSC preparations from bone marrow aspirates. Immunophenotyping, proliferation assays and NK cell cytotoxicity assays were employed to address the issue.

Results

While TStrC differed from MSC in terms of plasticity, they shared surface expression of CD105, CD73 and other markers used for MSC characterization. Furthermore, TStrC displayed a strong antiproliferative effect on peripheral blood mononuclear cells (PBMC) in coculture experiments similar to MSC. NK cell cytotoxicity was significantly impaired after co-culture with TStrC and expression of the activating NK cell receptors NKp44 and NKp46 was reduced.

Conclusions

Our data show that TStrC and MSC share important phenotypic and functional characteristics. The inhibitory effect of TStrC on PBMC and especially on NK cells may facilitate the immune evasion of paediatric tumours.
Antibiotic-free selection in E. coli: new considerations for optimal design and improved production
Authors: Peubez Isabelle , Chaudet Nicolas , Mignon Charlotte , Hild Géraldine , Husson Stéphanie , Courtois Virginie , De Luca Karelle , Speck Denis , Sodoyer Régis
Published: September 2010

Background

The increasing regulatory requirements to which biological agents are subjected will have a great impact in the field of industrial protein expression and production. There is an expectation that in a near future, there may be "zero tolerance" towards antibiotic-based selection and production systems. Besides the antibiotic itself, the antibiotic resistance gene is an important consideration. The complete absence of antibiotic-resistance gene being the only way to ensure that there is no propagation in the environment or transfer of resistance to pathogenic strains.

Results

In a first step, we have designed a series of vectors, containing a stabilization element allowing a complete elimination of antibiotics during fermentation. Vectors were further improved in order to include alternative selection means such as the well known poison/antidote stabilization system. Eventually we propose an elegant positive pressure of selection ensuring the elimination of the antibiotic-resistance gene through homologous recombination. In addition, we have shown that the presence of an antibiotic resistance gene can indirectly reduce the amount of expressed protein, since even in absence of selection pressure the gene would be transcribed and account for an additional stress for the host during the fermentation process.

Conclusions

We propose a general strategy combining plasmid stabilization and antibiotic-free selection. The proposed host/vector system, completely devoid of antibiotic resistance gene at the end of construction, has the additional advantage of improving recombinant protein expression and/or plasmid recovery.
MicroRNA roles in beta-catenin pathway
Authors: Huang Kai , Zhang Jun-Xia , Han Lei , You Yong-Ping , Jiang Tao , Pu Pei-Yu , Kang Chun-Sheng
Published: September 2010

β-catenin, a key factor in the Wnt signaling pathway, has essential functions in the regulation of cell growth and differentiation. Aberrant β-catenin signaling has been linked to various disease pathologies, including an important role in tumorigenesis. Here, we review the regulation of the Wnt signaling pathway as it relates to β-catenin signaling in tumorigenesis, with particular focus on the role of microRNAs. Finally, we discuss the potential of β-catenin targeted therapeutics for cancer treatment.
Genetic and diet effects on Ppar-α and Ppar-γ signaling pathways in the Berlin Fat Mouse Inbred line with genetic predisposition for obesity
Authors: Wagener Asja , Goessling Helge F , Schmitt Armin O , Mauel Susanne , Gruber Achim D , Reinhardt Richard , Brockmann Gudrun A
Published: September 2010

Background

The Berlin Fat Mouse Inbred (BFMI) line is a new mouse model for obesity, which was long-term selected for high fatness. Peroxisome proliferator-activated receptors (PPARs) are involved in the control of energy homeostasis, nutrient metabolism and cell proliferation. Here, we studied the expression patterns of the different Ppar genes and the genes in the PPAR pathway in the BFMI line in comparison to physiological changes.

Results

At the age of 10 weeks, the BFMI mice exhibited marked obesity with enlarged adipocytes and high serum triglycerides concentrations in comparison to the often used mouse line C57BL/6 (B6). Between these two lines, gene expression analyses revealed differentially expressed genes belonging to the PPAR pathway, in particular genes of the lipogenesis and the fatty acid transport.

Conclusion

Surprisingly, the Ppar-α gene expression was up-regulated in liver and Ppar-γ gene expression was down-regulated in the white adipose tissue, indicating the activation of a mechanism that counteracts the rise of obesity.
Profiling of cellular proteins in porcine reproductive and respiratory syndrome virus virions by proteomics analysis
Authors: Zhang Chengwen , Xue Chunyi , Li Yan , Kong Qingming , Ren Xiangpeng , Li Xiaoming , Shu Dingming , Bi Yingzuo , Cao Yongchang
Published: September 2010

Background

Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped virus, bearing severe economic consequences to the swine industry worldwide. Previous studies on enveloped viruses have shown that many incorporated cellular proteins associated with the virion's membranes that might play important roles in viral infectivity. In this study, we sought to proteomically profile the cellular proteins incorporated into or associated with the virions of a highly virulent PRRSV strain GDBY1, and to provide foundation for further investigations on the roles of incorporated/associated cellular proteins on PRRSV's infectivity.

Results

In our experiment, sixty one cellular proteins were identified in highly purified PRRSV virions by two-dimensional gel electrophoresis coupled with mass spectrometric approaches. The identified cellular proteins could be grouped into eight functional categories including cytoskeletal proteins, chaperones, macromolecular biosynthesis proteins, metabolism-associated proteins, calcium-dependent membrane-binding proteins and other functional proteins. Among the identified proteins, four have not yet been reported in other studied envelope viruses, namely, guanine nucleotide-binding proteins, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase, peroxiredoxin 1 and galectin-1 protein. The presence of five selected cellular proteins (i.e., β-actin, Tubulin, Annexin A2, heat shock protein Hsp27, and calcium binding proteins S100) in the highly purified PRRSV virions was validated by Western blot and immunogold labeling assays.

Conclusions

Taken together, the present study has demonstrated the incorporation of cellular proteins in PRRSV virions, which provides valuable information for the further investigations for the effects of individual cellular proteins on the viral replication, assembly, and pathogenesis.
Quantitative analysis of interferon alpha receptor subunit 1 and suppressor of cytokine signaling 1 gene transcription in blood cells of patients with chronic hepatitis C
Authors: Sedeño-Monge Virginia , Santos-López Gerardo , Rocha-Gracia Rosa C , Meléndez-Mena Daniel , Ramírez-Mata Alberto , Vallejo-Ruiz Verónica , Reyes-Leyva Julio
Published: September 2010

Background

Interferon (IFN)-α receptor 1 (ifnar1) and suppressor of cytokine signaling 1 (socs1) transcription levels were quantified in peripheral blood mononuclear cells (PBMC) of 59 patients infected with hepatitis C virus (HCV) and 17 non-infected individuals. Samples were obtained from patients infected with HCV that were either untreated or treated with IFN-α2 plus ribavirin for 1 year and divided into responders and non-responders based on viral load reduction 6 months after treatment. Ifnar1 and socs1 transcription was quantified by real-time RT-PCR, and the fold difference (2-ΔΔCT) with respect to hprt housekeeping gene was calculated.

Results

Ifnar1 transcription increased significantly in HCV-infected patients either untreated (3.26 ± 0.31), responders (3.1 ± 0.23) and non-responders (2.18 ± 0.23) with respect to non-infected individuals (1 ± 0.34; P = 0.005). Ifnar1 transcription increased significantly (P = 0.003) in patients infected with HCV genotypes 1a (4.74 ± 0.25) and 1b (2.81 ± 0.25) but not in 1a1b (1.58 ± 0.21). No association was found of Ifnar1 transcription with disease progress, initial viral load or other clinical factors. With respect to socs1 transcription, values were similar for non-infected individuals (1 ± 0.28) and untreated patients (0.99 ± 0.41) but increased in responders (2.81 ± 0.17) and non-responder patients (1.67 ± 0.41). Difference between responder and non-responder patients was not statistically significant. Socs1 transcription increased in patients infected with HCV genotypes 1a and 1b (2.87 ± 0.45 and 2.22 ± 0.17, respectively) but not in 1a1b (1.28 ± 0.40). Socs1 transcript was absent in three patients infected with HCV genotype 1b. A weak correlation between ifnar1 and socs1 transcription was found, when Spearman's correlation coefficient was calculated.

Conclusion

Our results suggest that HCV infection may up-regulate ifnar1 transcription. HCV genotypes differ in their capacity to affect ifnar1 and socs1 transcription, as well as in the ability to evade the antiviral response.
Coinfection with EBV/CMV and other respiratory agents in children with suspected infectious mononucleosis
Authors: Wang Xia , Yang Kun , Wei Cong , Huang Yuan , Zhao Dongchi
Published: September 2010

Background

Numerous studies have shown that Epstein-Barr virus (EBV) and cytomegalovirus (CMV) can infect immunocompetent patients simultaneously with other agents. Nonetheless, multiple infections with other agents in EBV/CMV-infected children have received little attention. We conducted a retrospective study of children with suspected infectious mononucleosis. Peripheral blood samples were analyzed by indirect immunofluorescence to detect EBV, CMV and other respiratory agents including respiratory syncytial virus; adenovirus; influenza virus types A and B; parainfluenza virus types 1, 2 and 3; Chlamydia pneumonia e and Mycoplasma pneumoniae. A medical history was collected for each child.

Results

The occurrence of multipathogen infections was 68.9%, 81.3% and 63.6% in the children with primary EBV, CMV or EBV/CMV, respectively, which was significantly higher than that in the past-infected group or the uninfected group (p < 0.001). Of the multipathogen-infected patients, the incidence of C. pneumoniae in children with primary infection was as high as 50%, significantly higher than in the other groups (p < 0.001). In the patients with multipathogen infection and EBV/CMV primary infection, fever, rash, lymphadenopathy, hepatomegaly, splenomegaly, atypical lymphocytes and abnormal liver function were more frequent and the length of hospital stay and duration of fever were longer than in other patients.

Conclusion

Our study suggests that there is a high incidence of multipathogen infections in children admitted with EBV/CMV primary infection and that the distribution of these pathogens is not random.
The ancient function of RB-E2F Pathway: insights from its evolutionary history
Authors: Cao Lihuan , Peng Bo , Yao Lei , Zhang Xinming , Sun Kuan , Yang Xianmei , Yu Long
Published: September 2010

Background

The RB-E2F pathway is conserved in most eukaryotic lineages, including animals and plants. E2F and RB family proteins perform crucial functions in cycle controlling, differentiation, development and apoptosis. However, there are two kinds of E2Fs (repressive E2Fs and active E2Fs) and three RB family members in human. Till now, the detail evolutionary history of these protein families and how RB-E2F pathway evolved in different organisms remain poorly explored.

Results

We performed a comprehensive evolutionary analysis of E2F, RB and DP (dimerization partners of E2Fs) protein family in representative eukaryotic organisms. Several interesting facts were revealed. First, orthologues of RB, E2F, and DP family are present in several representative unicellular organisms and all multicellular organisms we checked. Second, ancestral E2F, RB genes duplicated before placozoans and bilaterians diverged, thus E2F family was divided into E2F4/5 subgroup (including repressive E2Fs: E2F4 and E2F5) and E2F1/2/3 subgroup (including active E2Fs: E2F1, E2F2 and E2F3), RB family was divided into RB1 subgroup (including RB1) and RBL subgroup (including RBL1 and RBL2). Third, E2F4 and E2F5 share more sequence similarity with the predicted E2F ancestral sequence than E2F1, E2F2 and E2F3; E2F4 and E2F5 also possess lower evolutionary rates and higher purification selection pressures than E2F1, E2F2 and E2F3. Fourth, for RB family, the RBL subgroup proteins possess lower evolutionary rates and higher purification selection pressures compared with RB subgroup proteins in vertebrates,

Conclusions

Protein evolutionary rates and purification selection pressures are usually linked with protein functions. We speculated that function conducted by E2F4/5 subgroup and RBL subgroup proteins might mainly represent the ancient function of RB-E2F pathway, and the E2F1/2/3 subgroup proteins and RB1 protein might contribute more to functional diversification in RB-E2F pathway. Our results will enhance the current understanding of RB-E2F pathway and will also be useful to further functional studies in human and other model organisms.

Reviewers

This article was reviewed by Dr. Pierre Pontarotti, Dr. Arcady Mushegian and Dr. Zhenguo Lin (nominated by Dr. Neil Smalheiser).
Effect of etanercept in polymyalgia rheumatica: a randomized controlled trial
Authors: Kreiner Frederik , Galbo Henrik
Published: September 2010

Introduction

To elucidate in polymyalgia rheumatica (PMR) the role of tumor necrosis factor (TNF) α and the therapeutic potential of blockade with soluble TNF-α receptor, we carried out the first randomized controlled trial with etanercept in PMR.

Methods

Twenty newly diagnosed, glucocorticoid (GC) naïve patients with PMR and 20 matched non-PMR control subjects completed the trial. Subjects were randomized in a 1:1 ratio to monotherapy with etanercept (25 mg s.c. biweekly) or placebo (saline) for 14 days. Study outcomes were assessed at baseline and after 14 days. The primary outcome was the change in PMR activity score (PMR-AS). Secondary outcomes were: changes in erythrocyte sedimentation rate (ESR) and plasma levels of TNF-α and interleukin (IL) 6; patients' functional status (health assessment questionnaire) and cumulative tramadol intake during the trial.

Results

At baseline, plasma TNF-α was higher in patients than in controls (P < 0.05). The concentration always increased with etanercept treatment (P < 0.05). In patients, etanercept decreased PMR-AS by 24% (P = 0.011), reflecting significant improvements in shoulder mobility, physician's global assessment and C-reactive protein, and insignificant (P > 0.05) improvements in duration of morning stiffness and patient's assessment of pain. In parallel, ESR and IL-6 were reduced (P < 0.05). Placebo treatment did not change PMR-AS, ESR and IL-6 (P > 0.05). Functional status did not change and tramadol intake did not differ between patient groups. In controls, no changes occurred in both groups.

Conclusions

Etanercept monotherapy ameliorates disease activity in GC naïve patients with PMR. However, the effect is modest, indicating a minor role of TNF-α in PMR.

Trial registration

ClinicalTrials.gov (NCT00524381).
Genomic acquisition of a capsular polysaccharide virulence cluster by non-pathogenic Burkholderia isolates
Authors: Sim Bernice Meng Qi , Chantratita Narisara , Ooi Wen Fong , Nandi Tannistha , Tewhey Ryan , Wuthiekanun Vanaporn , Thaipadungpanit Janjira , Tumapa Sarinna , Ariyaratne Pramila , Sung Wing-Kin , Sem Xiao Hui , Chua Hui Hoon , Ramnarayanan Kalpana , Lin Chi Ho , Liu Yichun , Feil Edward J , Glass Mindy B , Tan Gladys , Peacock Sharon J , Tan Patrick
Published: September 2010

Background

Burkholderia thailandensis is a non-pathogenic environmental saprophyte closely related to Burkholderia pseudomallei, the causative agent of the often fatal animal and human disease melioidosis. To study B. thailandensis genomic variation, we profiled 50 isolates using a pan-genome microarray comprising genomic elements from 28 Burkholderia strains and species.

Results

Of 39 genomic regions variably present across the B. thailandensis strains, 13 regions corresponded to known genomic islands, while 26 regions were novel. Variant B. thailandensis isolates exhibited isolated acquisition of a capsular polysaccharide biosynthesis gene cluster (B. pseudomallei-like capsular polysaccharide) closely resembling a similar cluster in B. pseudomallei that is essential for virulence in mammals; presence of this cluster was confirmed by whole genome sequencing of a representative variant strain (B. thailandensis E555). Both whole-genome microarray and multi-locus sequence typing analysis revealed that the variant strains formed part of a phylogenetic subgroup distinct from the ancestral B. thailandensis population and were associated with atypical isolation sources when compared to the majority of previously described B. thailandensis strains. In functional assays, B. thailandensis E555 exhibited several B. pseudomallei-like phenotypes, including colony wrinkling, resistance to human complement binding, and intracellular macrophage survival. However, in murine infection assays, B. thailandensis E555 did not exhibit enhanced virulence relative to other B. thailandensis strains, suggesting that additional factors are required to successfully colonize and infect mammals.

Conclusions

The discovery of such novel variant strains demonstrates how unbiased genomic surveys of non-pathogenic isolates can reveal insights into the development and emergence of new pathogenic species.
Temozolomide Versus Procarbazine, Lomustine, and Vincristine in Recurrent High-Grade Glioma
Authors: Brada Michael , Stenning Sally , Gabe Rhian , Thompson Lindsay C. , Levy David , Rampling Roy , Erridge Sara , Saran Frank , Gattamaneni Rao , Hopkins Kirsten , Beall Sarah , Collins V. Peter , Lee Siow-Ming
Published: September 2010

Purpose

Temozolomide (TMZ) is an alkylating agent licensed for treatment of high-grade glioma (HGG). No prospective comparison with nitrosourea-based chemotherapy exists. We report, to our knowledge, the first randomized trial of procarbazine, lomustine, and vincristine (PCV) versus TMZ in chemotherapy-naive patients with recurrent HGG.

Patients and Methods

Four hundred forty-seven patients were randomly assigned to PCV (224 patients) or TMZ (sub–random assignment: TMZ-5 [200 mg/m2 for 5 days, 112 patients] or TMZ-21 [100 mg/m2 for 21 days, 111 patients]) for up to 9 months or until progression. The primary outcomes were survival (PCV v TMZ) and 12-week progression-free survival (PFS; TMZ-5 v TMZ-21). This study is registered as ISRCTN83176944.

Results

Percentages of patients completing 9 months of treatment in the PCV, TMZ-5, and TMZ-21 arms were 17%, 26%, and 13%, respectively. Major toxicity was similar across all three groups. With a median follow-up time of 12 months and 382 deaths, there was no clear survival benefit when comparing PCV with TMZ (hazard ratio [HR], 0.91; 95% CI, 0.74 to 1.11; P = .350). For TMZ-5 versus TMZ-21, 12-week PFS rates were similar (63.6% and 65.7%, respectively; P = .745), but TMZ-5 improved overall PFS (HR, 1.38; 95% CI, 1.05 to 1.82; P = .023), survival (HR, 1.32; 95% CI, 0.99 to 1.75; P = .056), and global quality of life (49% v 19% improved > 10 points at 6 months, respectively; P = .005).

Conclusion

Although TMZ (both arms combined) did not show a clear benefit compared with PCV, comparison of the TMZ schedules demonstrated that the 21-day schedule was inferior to the 5-day schedule in this setting. This challenges the current understanding of increasing TMZ dose-intensity by prolonged scheduling.
Survival Outcomes and Prognostic Factors in Mycosis Fungoides/Sézary Syndrome: Validation of the Revised International Society for Cutaneous Lymphomas/European Organisation for Research and Treatment of Cancer Staging Proposal
Authors: Agar Nita Sally , Wedgeworth Emma , Crichton Siobhan , Mitchell Tracey J. , Cox Michael , Ferreira Silvia , Robson Alistair , Calonje Eduardo , Stefanato Catherine M. , Wain Elizabeth Mary , Wilkins Bridget , Fields Paul A. , Dean Alan , Webb Katherine , Scarisbrick Julia , Morris Stephen , Whittaker Sean J.
Published: September 2010

Purpose

We have analyzed the outcome of mycosis fungoides (MF) and Sézary syndrome (SS) patients using the recent International Society for Cutaneous Lymphomas (ISCL)/European Organisation for Research and Treatment of Cancer (EORTC) revised staging proposal.

Patients and Methods

Overall survival (OS), disease-specific survival (DSS), and risk of disease progression (RDP) were calculated for a cohort of 1,502 patients using univariate and multivariate models.

Results

The mean age at diagnosis was 54 years, and 71% of patients presented with early-stage disease. Disease progression occurred in 34%, and 26% of patients died due to MF/SS. A significant difference in survival and progression was noted for patients with early-stage disease having patches alone (T1a/T2a) compared with those having patches and plaques (T1b/T2b). Univariate analysis established that (1) advanced skin and overall clinical stage, increased age, male sex, increased lactate dehydrogenase (LDH), and large-cell transformation were associated with reduced survival and increased RDP; (2) hypopigmented MF, MF with lymphomatoid papulosis, and poikilodermatous MF were associated with improved survival and reduced RDP; and (3) folliculotropic MF was associated with an increased RDP. Multivariate analysis established that (1) advanced skin (T) stage, the presence in peripheral blood of the tumor clone without Sézary cells (B0b), increased LDH, and folliculotropic MF were independent predictors of poor survival and increased RDP; (2) large-cell transformation and tumor distribution were independent predictors of increased RDP only; and (3) N, M, and B stages; age; male sex; and poikilodermatous MF were only significant for survival.

Conclusion

This study has validated the recently proposed ISCL/EORTC staging system and identified new prognostic factors.
Phase II Study of Dasatinib in Patients With Advanced Non–Small-Cell Lung Cancer
Authors: Johnson Faye M. , Bekele B. Nebiyou , Feng Lei , Wistuba Ignacio , Tang Xi Ming , Tran Hai T. , Erasmus Jeremy J. , Hwang Li-Ling , Takebe Naoko , Blumenschein George R. , Lippman Scott M. , Stewart David J.
Published: September 2010

Purpose

Src family kinases (SFKs) promote cancer progression and are commonly expressed in non–small-cell lung cancer (NSCLC), but the clinical effects of SFK inhibition in NSCLC are unknown. We conducted a phase II trial of the SFK inhibitor dasatinib for advanced NSCLC. We tested the hypotheses that the activation of epidermal growth factor receptor (EGFR) or SFK or modulation of serum cytokines may predict a response to dasatinib.

Patients and Methods

Patients received dasatinib as first-line therapy. Response was measured by tumor size on computed tomography scans and by metabolic activity on positron emission tomography scans. Tissue samples taken before patients received dasatinib were tested for EGFR and Kras mutation and phosphorylated SFK expression.

Results

Thirty-four patients were enrolled. The overall disease control rate (partial responses plus stable disease) for dasatinib was 43%. One patient had a partial response to therapy. Eleven patients (32%) had a metabolic response to dasatinib. SFK activation and EGFR and Kras mutations in tumor tissue did not predict response to dasatinib. Significant toxicities included fatigue and dyspnea. The presence of a pleural effusion before dasatanib therapy predicted the development of a clinically significant effusion during therapy.

Conclusion

Dasatinib as a single agent had modest clinical activity that was lower than that generally observed in patients with NSCLC who receive chemotherapy. Pleural effusion was an expected and problematic toxicity that was successfully treated with steroids, diuretics, and dose interruptions. Marked activity in one patient and prolonged stable disease in four others suggested a potential subpopulation of patients with dasatinib-sensitive NSCLC.
Summer Is Over
Authors: Black John
Published: September 2010

The College exists to set standards for surgical practice and one of the areas in which we do this is the consultant appointment process. The College regional specialty professional advisor must approve the job description for the post before it can be advertised and there must be a College assessor at the appointment committee. The reason for this is to provide external objective assurance that the full range of working arrangements for the individual are compatible with proper consultant practice and that the person appointed is trained appropriately for the post. Foundation trusts are technically not bound to follow the NHS regulations but to date have continued to involve the College in the traditional way.
The Impact of ISTCS – A Clinician's Perspective (Questionnaire Study)
Authors: Mendis D , Hawrani A
Published: September 2010

Independent sector treatment centres and their effect on the NHS remain a controversial aspect of healthcare delivery. This postal questionnaire study aims to identify the general attitude among surgical consultants about their effects, specifically on NHS workload, departmental/trust finances, training opportunities and case mix. NHS hospitals within five miles of an ISTC offering day case/inpatient services were targeted.
The Concept of Surgical Assessment: Part 2 – Available Tools
Authors: Okoro T , Sirianni C , Brigden D
Published: September 2010

In the past surgical training has been based on many years and long hours of work experience together with examinations, which have been mainly knowledge-based. The reduction in the clinical experience of trainees caused by shifts and the European Working Time Regulations means that new ways are needed to facilitate training and to ensure that competence is achieved. In this paper the current methods of assessment of surgical ability in the UK will be outlined. Other tools available that have evidence to support their use will also be described.
Controls for Immunocytochemistry
Authors: Burry Richard W.
Published: September 2010

Immunocytochemistry is a highly productive method in biomedical research used to identify proteins and other macromolecules in tissues and cells. Control samples are required to show label localization is correct, but the understanding and use of immunocytochemistry controls have been inconsistent. A new classification of immunocytochemical controls is proposed that will help in understanding this most important component of the experiment. The three types of controls required for immunocytochemistry are primary antibody controls that show the specificity of the primary antibody binding to the antigen, secondary antibody controls that show the label is specific to the primary antibody, and label controls that show the labeling is the result of the label added and not the result of endogenous labeling. Publications containing immunocytochemical results must give details of how these controls were performed.
Association of SRD5A2 Variants and Serum Androstane-3α,17β-Diol Glucuronide Concentration in Chinese Elderly Men
Authors: Jiang Jieying , Tang Nelson LS , Ohlsson Claes , Eriksson Anna L , Vandenput Liesbeth , Liao ChenDi , Wang Xingyan , Chan Frank WK , Kwok Anthony , Orwoll Eric , Kwok Timothy CY , Woo Jean , Leung Ping Chung
Published: September 2010

BACKGROUND

Results of recent studies have demonstrated that genetic variants of the enzyme steroid 5α reductase type II (SRD5A2) are associated with serum concentrations of major androgen metabolites such as conjugates of androstane-3α,17β-diol-glucuronide (3α-diol-G). However, this association was not consistently found among different ethnic groups. Thus, we aimed to determine whether the association with SRD5A2 genetic variations exists in a cohort of healthy Chinese elderly men, by examining 2 metabolite conjugates: androstane-3α,l7β-diol-3-glucuronide (3α-diol-3G) and androstane-3α,17β-diol-17-glucuronide (3α-diol-17G).

METHODS

We used GC-MS and LC-MS to measure serum sex steroid concentrations, including testosterone and dihydrotestosterone, and 3α-diol-3G and 3α-diol-17G in 1182 Chinese elderly men age 65 and older. Genotyping of the 3 SRD5A2 tagSNPs [rs3731586, rs12470143, and rs523349 (V89L)] was performed by using melting-temperature–shift allele-specific PCR.

RESULTS

The well-described SRD5A2 missense variant rs523349 (V89L) was modestly associated with the 3α-diol-17G concentration (P = 0.040). On the other hand, SNP rs12470143 was found to be significantly correlated with 3α-diol-3G concentration (P = 0.021). Results of haplotype analysis suggested that the presence of an A-C-G haplotype leads to an increased 3α-diol-3G concentration, a finding consistent with results of single SNP analysis.

CONCLUSIONS

The genetic variation of SRD5A2 is associated with circulating 3α-diol-3G and 3α-diol-17G concentrations in Chinese elderly men. In addition, we showed that SRD5A2 haplotypic association, rather than a single SNP alone, might be a better predictor of the 3α-diol-G concentration. Thus, the effect of either the haplotype itself or of other ungenotyped SNPs in linkage disequilibrium with the haplotype is responsible for the interindividual variation of 3α-diol-G.
Microsatellite Instability Detection by High-Resolution Melting Analysis
Authors: Janavicius Ramunas , Matiukaite Dovile , Jakubauskas Arturas , Griskevicius Laimonas
Published: September 2010

BACKGROUND

Microsatellite instability (MSI) is an important marker for screening for hereditary nonpolyposis colorectal cancer (Lynch syndrome) as well as a prognostic and predictive marker for sporadic colorectal cancer (CRC). The mononucleotide microsatellite marker panel is a well-established and superior alternative to the traditional Bethesda MSI analysis panel, and does not require testing for corresponding normal DNA. The most common MSI detection techniques—fluorescent capillary electrophoresis and denaturing HPLC (DHPLC)—both have advantages and drawbacks. A new high-resolution melting (HRM) analysis method enables rapid identification of heteroduplexes in amplicons by their lower thermal stability, a technique that overcomes the main shortcomings of capillary electrophoresis and DHPLC.

METHODS

We investigated the straightforward application of HRM for the detection of MSI in 70 archival CRC samples. HRM analysis for 2 MSI markers (BAT25 and BAT26) was evaluated, and 2 different HRM-enabled instruments were compared—the LightCycler® 480 (Roche Diagnostics) and the LightScannerTM (Idaho Technology). We also determined the analytical sensitivity and specificity of the HRM assay on both instruments using 11 known MSI-positive and 54 microsatellite-stable CRC samples.

RESULTS

All MSI-positive samples were detected on both instruments (100% analytical sensitivity). The LightScanner performed better for analytical specificity, giving a combined specificity value of 99.1% compared with 92.3% on the LightCycler 480.

CONCLUSIONS

We expanded the application of the HRM analysis method as an effective MSI detection technique for clinical samples.
Long Life Fatigue Fracture Induced by Interior Inclusions for High Carbon Chromium Bearing Steels Under Rotating Bending
Authors: Oguma N. , Lian B. , Sakai T. , Watanabe K. , Odake Y.
Published: September 2010

The characteristics of the long life fatigue fractures induced by interior nonmetallic inclusions for two types of high carbon chromium bearing steels were experimentally examined under rotating bending. Specimens were machined from the round bars with different diameters. One type was cut from the central area of a thin bar; the other type was cut from the medium area of the radius of a thick bar. As a result of the fatigue tests, the fatigue life of the latter type specimen was longer in the long life region. Crack initiation sites on fracture surfaces of all the failed specimens were observed by means of a scanning electron microscope. The number of alumina inclusions was decreased, and the average size of titanium nitride inclusions on fracture surfaces was smaller for the latter type specimens. And a characteristic rough surface of fine granular area (FGA) was observed in a circle around the inclusion on the fracture surface. The size of FGA tended to increase with an increase of the fatigue life and a decrease of the stress amplitude at the position of the inclusion. From a view point of fracture mechanics, the stress intensity factor range at the front of FGA, ⊿KFGA, was calculated for each specimen by using √area model. Values of ⊿KFGA of both type specimens were almost constant regardless of the stress amplitude at the site of inclusion, and the average value was about 5 MPa√m. That is, it seems that the fatigue life of the specimens cut from the thick bar was improved by downsizing the inclusion area on the fracture surface.
Systemic Hypothermia After Neonatal Encephalopathy: Outcomes of neo.nEURO.network RCT
Authors: Simbruner Georg , Mittal Rashmi A. , Rohlmann Friederike , Muche Rainer ,   neo.nEURO.network Trial Participants
Published: September 2010

OBJECTIVE:

Mild hypothermia after perinatal hypoxic-ischemic encephalopathy (HIE) reduces neurologic sequelae without significant adverse effects, but studies are needed to determine the most-efficacious methods.

METHODS:

In the neo.nEURO.network trial, term neonates with clinical and electrophysiological evidence of HIE were assigned randomly to either a control group, with a rectal temperature of 37°C (range: 36.5–37.5°C), or a hypothermia group, cooled and maintained at a rectal temperature of 33.5°C (range: 33–34°C) with a cooling blanket for 72 hours, followed by slow rewarming. All infants received morphine (0.1 mg/kg) every 4 hours or an equivalent dose of fentanyl. Neurodevelopmental outcomes were assessed at the age of 18 to 21 months. The primary outcome was death or severe disability.

RESULTS:

A total of 129 newborn infants were enrolled, and 111 infants were evaluated at 18 to 21 months (53 in the hypothermia group and 58 in the normothermia group). The rates of death or severe disability were 51% in the hypothermia group and 83% in the normothermia group (P = .001; odds ratio: 0.21 [95% confidence interval [CI]: 0.09–0.54]; number needed to treat: 4 [95% CI: 3–9]). Hypothermia also had a statistically significant protective effect in the group with severe HIE (n = 77; P = .005; odds ratio: 0.17 [95% CI: 0.05–0.57]). Rates of adverse events during the intervention were similar in the 2 groups except for fewer clinical seizures in the hypothermia group.

CONCLUSION:

Systemic hypothermia in the neo.nEURO.network trial showed a strong neuroprotective effect and was effective in the severe HIE group.
Racial Differences in the Effects of Postnatal Environmental Tobacco Smoke on Neurodevelopment
Authors: Xu Xiaohui , Cook Robert L. , Ilacqua Vito A. , Kan Haidong , Talbott Evelyn O.
Published: September 2010

OBJECTIVES:

We used the 2001–2004 National Health and Nutrition Examination Survey to examine the association between postnatal environmental tobacco smoke exposure, measured as serum cotinine levels, and attention-deficit/hyperactivity disorder (ADHD) among children 4 to 15 years of age. We further investigated the interactions of race and serum cotinine levels with ADHD.

METHODS:

Logistic regression models were used to evaluate associations.

RESULTS:

This study found that the prevalence of ADHD increased as blood cotinine levels increased. The effects of blood cotinine levels on ADHD differed according to race. Compared with children of the same racial group with the lowest blood cotinine levels, the odds ratios were 2.72 (95% confidence interval: 1.25–5.93) for Mexican American children and 5.32 (95% confidence interval: 1.55–18.3) for children in other racial groups with the highest blood cotinine levels, with controlling for the effect of maternal smoking during pregnancy. However, no significant associations between blood cotinine levels and ADHD were observed among non-Hispanic white or non-Hispanic black children.

CONCLUSIONS:

The findings of this study underscore the possibility of racial disparities in the effects of environmental tobacco smoke on behavioral problems in children. These findings warrant further investigation.
Visualization of U and M genome chromosomes by multicolour genomic in situ hybridization in Aegilops biuncialis and Triticum aestivum-Ae. biuncialis amphiploids
Authors: Molnár I. , Molnár-Láng M.
Published: September 2010

The multicolour genomicin situhybridization (mcGISH) method was improved in order to visualize the Uband Mbgenomes ofAegilops biuncialisVis. (2n=4x=28, UbUbMbMb). Hybridization probes prepared from the diploid U and M genome donors,Ae. umbellulataandAe. comosa, resulted in clear hybridization signals on the U and M chromosomes inAe. biuncialis. The random primed labelling method made it possible to decrease the blocking ratio to 1:30. McGISH allowed the simultaneous discrimination of individualAe. biuncialisgenomes and wheat chromosomes in γ-irradiatedTriticum aestivum-Ae. biuncialisamphiploids (2n=70; AABBDDUbUbMbMb). Dicentric chromosomes, terminal and interstitial translocations and centric fusions were detected in the irradiated generation. The irradiation-induced wheat-Ae. biuncialisintergenomic translocations will facilitate the successful introgression of useful agronomic traits into bread wheat.
Breeding Rosa taxa native to the Carpathian Basin for fruit purposes — fruit quality
Authors: Kovács S. , Udvardy L. , Tóth M.
Published: September 2010

The aim of dogrose breeding for fruit purposes is to select genotypes suitable for cultivation and to produce new genotypes by crossing. Physical and chemical analyses, prospective genotypes have been developed from
R. inodora
,
R. corymbifera, R. rubiginosa
and
R. canina
varieties.In the course of the investigations, the highest vitamin C content was found in the hips of
R. inodora
and
R. rubiginosa
. The glucose and fructose contents ranged from 9.57–13.36 g/100 g, averaged over several years. The amounts of these two carbohydrates were equal, or in some taxa (e.g.
R. corymbifera, R. canina
Sz3) the fructose content was higher.The glucose, fructose and vitamin C contents changed at different rates in each taxon during ripening. The results showed that the fructose content reached its peak a week earlier than the glucose content. The vitamin C content of morphological varieties of
R. canina
did not change substantially during ripening.
SPME-GC/MS identification of aroma compounds in rose flowers
Authors: Héthelyi É. , Szarka S. , Lemberkovics É. , Szőke É.
Published: September 2010

The content and composition of active ingredients and essential oils in medicinal and aromatic plants have been studied for several decades. The volatile compounds in essential oils have been analysed routinely using gas chromatography (GC) since 1966, and with GC coupled to mass spectrometric detection (GC/MS) since 1978.The 13 rose varieties selected for chemical analysis varied for colour, shape and fragrance. The static headspace solid phase microextraction (sHS-SPME) technique recently developed for sample preparation and sample enrichment was used to study the volatile aromatic components.The main volatile compound of a sweet-smelling purple rose was found to be phenyl ethyl alcohol (33–52%). The phenyl ethyl alcohol content of fragrant rose flowers with blackish-purple petals increased continuously from early summer to late autumn (from 17 to 70 %). The dominant aromatic components of the yellow, orange and pink rose flowers were hexanol, hexenyl acetate and benzyl alcohol. Phenyl ethyl alcohol and orcinol dimethyl ether were the main constituents of the fragrant pink and white rose varieties. Methyl vinyl anisol and orcinol dimethyl ether were dominant in rose flowers with beige petals. In summary, it can be concluded that the SPME-GC/MS method is suitable for the characterization of rose varieties and for the chemical analysis of aromatic volatile compounds.
Different responses of sensitive and resistant apricot genotypes to artificial Monilia laxa (Aderh. & Ruhl.) infection
Authors: Gutermuth Á. , Lendvay B. , Pedryc A.
Published: September 2010

Blossom blight caused byMonilia laxa(Ehr.) is the most important fungal disease in Hungarian apricot orchards. The cultivars traditionally grown in the country are susceptible toMonilia laxa(Ehr.) to various extents. In this study the shoots of one tree each of the varieties Zard and Korai Zamatos and 48 hybrids from their cross were artificially infectedin vivowithMonilia laxa(Ehr.). The results indicated that when artificial infections are made to evaluate pathogen resistance, this should be carried out on one-year-old shoots, since this is the natural infection point ofMonilia. It also appears that, due to the great variability in the size of destroyed tissues, the microscopic analysis of the infections could provide a more reliable evaluation of progeny resistance than comparing the sizes of destroyed shoot areas.
Goals, present position and prospects of forage sorghum breeding in Hungary
Authors: Pál M. , Rajki E.
Published: September 2010

Forage sorghum species, comprising grain and silage sorghum (S
orghum bicolor)
and sudangrass
(Sorghum sudanense
), are considered the most drought-tolerant field crops in Hungary (Siklósiné Rajki, 1997). Several features, such as its tropical origin, deep voluminous roots, and thick waxy leaves and stem, enable the plants to survive periods with insufficient rainfall (Wall and Ross, 1970). Owing to its large roots, the crop shows wide adaptability to soils with moderate or low fertility. Though sorghum is used mainly for forage in Hungary, it is a very important source of bioenergy in other countries. Hungary lies in the northern part of the European sorghum belt, so the emphasis in the breeding programme is to develop early or mid-early forage sorghum hybrids adapted to Hungarian climatic conditions and soil properties, to introduce them into commercial production and to make them available for farmers.
Characterization of wheat-barley introgression lines for drought tolerance
Authors: Hoffmann B. , Aranyi N. , Molnár-Láng M.
Published: September 2010

The safety of wheat production in Hungary requires the propagation of droughttolerant cultivars because of the regular occurrence of water deficiency. Hybridization between related species makes it possible to transfer desirable traits from one species to another. Introgression lines developed from wheat/barley hybrids were investigated together with the parental wheat and barley cultivars to determine how the added barley chromosome (segment) influences drought tolerance in wheat. The plants were grown in the field at the UP Georgikon Faculty, Keszthely. Sowing and harvest were done by hand. Half the length of the 12 m rows was covered with a plastic rain shelter on 2
nd
April (EC: 30–31) to protect the plants from rain, resulting in a 163 mm difference in water supplies between the control (not covered) and stressed (covered) treatments. Data were obtained for anthesis and maturity date, plant height, root/shoot ratio, leaf water potential, grain yield and grain yield components. The plants adapted to water deficiency by increasing the root/shoot ratio and decreasing the water potential and the duration of grain filling. The grain yield was reduced by 12%, averaged over the genotypes, mainly due to a decrease in the number of spikes per plant.
Tradition, quality and biotechnology in Hungarian spice pepper ( Capsicum annuum L.) breeding
Authors: Pauk J. , Lantos C. , Somogyi G. , Vági P. , Ábrahám Táborosi Z. , Gémes Juhász A. , Mihály R. , Kristóf Z. , Somogyi N. , Tímár Z.
Published: September 2010

Spice pepper production has a history of almost 300 years in the southern part of Hungary. In this study the results of two biotechnological improvements are summarized. Anther and isolated microspore culture techniques were improved to release haploid and doubled haploid (DH) lines for spice pepper breeding. Both the anther and isolated microspore culture methods were successfully used in spice pepper haploid production. Microspore culture-derived structures were analysed to identify their different parts. Green plantlets were regenerated from embryos derived from both anther and microspore cultures. Their doubled haploid analogues were integrated into Hungarian spice pepper hybrid seed breeding programmes. One hybrid, Sláger, was released as a new genotype for spice pepper production in 2008 and two hybrid candidates (Délibáb and Bolero) are now being tested in official trials.
In silico analysis of a putative SPIRAL gene related to strawberry ripening
Authors: Polgári D. , Kalapos B. , Tisza V. , Kovács L. , Kerti B. , Heszky L. , Kiss E.
Published: September 2010

The aim of this study was to characterize a gene associated with ripening in strawberry, a non-climacteric fruit. Differently expressed transcripts of candidate genes functioning in fruit development and ripening were identified from strawberry (Fragaria×ananassaDuch.) in four ripening stages using the cDNA-AFLP method. The cDNA fragment designated C11M32M003 was selected from the putative ripening-related genes for further analysis. This transcript accumulated in the green receptacle, and the achene, but gene expression decreased in both tissues in parallel with the progress of ripening (Balogh, 2006).In silicoanalysis revealed that both the cDNA-AFLP fragment (C11M32M003) and the full-length cDNA AY695666 showed over 60% homology at the nucleotide level with two gene groups found in various plant species, includingArabidopsis thaliana. One of the candidate groups consisted ofNITRILASEsequences thought to be related to auxin biosynthesis. As an alternative, a lesser known gene group namedSPIRALwas suggested. The results of the detailed bioinformatic comparisons presented in this paper prove that the strawberry sequence analysed belongs to theSPIRALgene family.