bluuebunny/crossref_metadata_2025 huggingface dataset:
Initial experience of transumbilical laparoendoscopic single-site surgery of partial adrenalectomy in patient with aldosterone-producing adenoma
Authors: Yuge Kazuyuki , Miyajima Akira , Hasegawa Masanori , Miyazaki Yasumasa , Maeda Takahiro , Takeda Toshikazu , Takeda Ayano , Miyashita Kazutoshi , Kurihara Isao , Shibata Hirotaka , Kikuchi Eiji , Oya Mototsugu
Published: November 2010

Background

Laparoscopic single-site surgery has recently emerged in the field of urology and this minimally-invasive surgery has resulted in a further reduction in morbidity compared with traditional laparoscopy. We present our initial experience with laparoendoscopic single-site surgery of partial adrenalectomy (LESS-PA) to treat aldosterone-producing adenomas.

Case presentation

A 60-year-old woman was diagnosed with aldosterone-producing macroadenomas in the left adrenal and aldosterone-producing microadenomas in the right adrenal. A two-step operation was planned. The first step involved transumbilical LESS-PA for the left adrenal tumors. A multichannel port was inserted through the center of the umbilicus and the left adrenal gland was approached using bent instruments according to standard traditional laparoscopic procedures. The tumors were resected using an ultrasonic scalpel, and the resected site was coagulated using a vessel sealing instrument and then sealed with fibrin glue. Operative time was 123 minutes and blood loss was minimal. The patient was discharged from hospital within 72 hours. Her right adrenal microadenomas will be treated in the next several months.

Conclusions

Although our experience is limited, LESS-PA appears to be safe and feasible for treating aldosterone-producing adenomas. More cases and comparisons with the multiport technique are needed before drawing any definite conclusions concerning the technique.
Echinacea-induced cytosolic Ca2+ elevation in HEK293
Authors: Wu Lankun , Rowe Eric W , Jeftinija Ksenija , Jeftinija Srdija , Rizshsky Ludmila , Nikolau Basil J , McKay Jodi , Kohut Marian , Wurtele Eve Syrkin
Published: November 2010

Background

With a traditional medical use for treatment of various ailments, herbal preparations of Echinacea are now popularly used to improve immune responses. One likely mode of action is that alkamides from Echinacea bind to cannabinoid type 2 (CB2) receptors and induce a transient increase in intracellular Ca2+. Here, we show that unidentified compounds from Echinacea purpurea induce cytosolic Ca2+ elevation in non-immune-related cells, which lack CB2 receptors and that the Ca2+ elevation is not influenced by alkamides.

Methods

A non-immune human cell line, HEK293, was chosen to evaluate E. purpurea root extracts and constituents as potential regulators of intracellular Ca2+ levels. Changes in cytosolic Ca2+ levels were monitored and visualized by intracellular calcium imaging. U73122, a phospholipase C inhibitor, and 2-aminoethoxydiphenyl borate (2-APB), an antagonist of inositol-1,4,5-trisphosphate (IP3) receptor, were tested to determine the mechanism of this Ca2+ signaling pathway. E. purpurea root ethanol extracts were fractionated by preparative HPLC, screened for bioactivity on HEK293 cells and by GC-MS for potential constituent(s) responsible for this bioactivity.

Results

A rapid transient increase in cytosolic Ca2+ levels occurs when E. purpurea extracts are applied to HEK293 cells. These stimulatory effects are phospholipase C and IP3 receptor dependent. Echinacea-evoked responses could not be blocked by SR 144528, a specific CB2 receptor antagonist, indicating that CB2 is not involved. Ca2+ elevation is sustained after the Echinacea-induced Ca2+ release from intracellular Ca2+ stores; this longer-term effect is abolished by 2-APB, indicating a possible store operated calcium entry involvement. Of 28 HPLC fractions from E. purpurea root extracts, six induce cytosolic Ca2+ increase. Interestingly, GC-MS analysis of these fractions, as well as treatment of HEK293 cells with known individual and combined chemicals, indicates the components thought to be responsible for the major immunomodulatory bioactivity of Echinacea do not explain the observed Ca2+ response. Rather, lipophilic constituents of unknown structures are associated with this bioactivity.

Conclusions

Our data indicate that as yet unidentified constituents from Echinacea stimulate an IP3 receptor and phospholipase C mediation of cytosolic Ca2+ levels in non-immune mammalian cells. This pathway is distinct from that induced in immune associated cells via the CB2 receptor.
Lipid control and use of lipid-regulating drugs for prevention of cardiovascular events in Chinese type 2 diabetic patients: a prospective cohort study
Authors: Ting Rose ZW , Yang Xilin , Yu Linda WL , Luk Andrea OY , Kong Alice PS , Tong Peter CY , So Wing-Yee , Chan Juliana CN , Ma Ronald CW
Published: November 2010

Background

Dyslipidaemia is an important but modifiable risk factor of cardiovascular disease (CVD) in type 2 diabetes. Yet, the effectiveness of lipid regulating drugs in Asians is lacking. We examined the effects of lipid control and treatment with lipid regulating drugs on new onset of CVD in Chinese type 2 diabetic patients.

Methods

In this prospective cohort consisting of 4521 type 2 diabetic patients without history of CVD and naïve for lipid regulating treatment recruited consecutively from 1996 to 2005, 371 developed CVD after a median follow-up of 4.9 years. We used Cox proportional hazard regression to obtain the hazard ratios (HR) of lipids and use of lipid regulating drugs for risk of CVD.

Results

The multivariate-adjusted HR (95% confidence interval) of CVD in patients with high LDL-cholesterol (≥ 3.0 mmol/L) was 1.36 (1.08 - 1.71), compared with lower values. Using the whole range value of HDL-cholesterol, the risk of CVD was reduced by 41% with every 1 mmol/L increase in HDL-cholesterol. Plasma triglyceride did not predict CVD. Statins use was associated with lower CVD risk [HR = 0.66 (0.50 - 0.88)]. In sub-cohort analysis, statins use was associated with a HR of 0.60 (0.44 - 0.82) in patients with high LDL-cholesterol (≥ 3.0 mmol/L) and 0.49 (0.28 - 0.88) in patients with low HDL-cholesterol. In patients with LDL-cholesterol < 3.0 mmol/L, use of fibrate was associated with HR of 0.34 (0.12 - 1.00). Only statins were effective in reducing incident CVD in patients with metabolic syndrome [(HR = 0.58(0.42--0.80)].

Conclusions

In Chinese type 2 diabetic patients, high LDL-cholesterol and low HDL-cholesterol predicted incident CVD. Overall, patients treated with statins had 40-50% risk reduction in CVD compared to non-users.
Epigenetic silencing of MAL, a putative tumor suppressor gene, can contribute to human epithelium cell carcinoma
Authors: Cao Wei , Zhang Zhi-yuan , Xu Qin , Sun Qiang , Yan Ming , Zhang Jun , Zhang Ping , Han Ze-guang , Chen Wan-tao
Published: November 2010

Background

To identify new and useful candidate biomarkers in head and neck squamous cell carcinoma (HNSCC), we performed a genome-wide survey and found that Myelin and lymphocyte-associated protein (MAL) was a gene that was markedly down-regulated in HNSCC. Hence, we investigated the mechanism ofMALsilencing and the effects ofMALon the proliferation, invasion, and apoptotic potential in HNSCC.

Results

MALwas significantly down-regulated in 91.7% of HNSCC specimens at the mRNA level as compared with adjacent normal tissues (P= 0.0004). Moreover, the relative transcript levels of theMALgene were remarkably decreased by five-fold in nine HNSCC cell lines as compared with normal head and neck epithelium cells.MALgene expression was restored in 44%, 67%, and 89% in HNSCC cell lines treated with TSA, 5-Aza-dC, and TSA plus 5-Aza-dC, respectively. Furthermore, bisulfate-treated DNA sequencing demonstrated that the two CpG islands (that is, M1and M2) located inMALpromoter region were completely methylated in the HNSCC cell lines (CpG methylated ratio was more than 90%), and only one CpG island (that is, M1) was partially methylated in HNSCC tissues (CpG methylated ratio between 20% and 90%). A significant reduction in cell proliferation and a change in the cell cycle profile were also observed inMALtransfectants. Matrigel assay demonstrated that the invasiveness of HNSCC cells significantly decreased. A significant increase in the population of apoptotic cells was observed inMALtransfected cells. The exogenous expression of theMALgene suppressed malignant phenotypes, while the cell death induced byMALgene transfer was a result of apoptosis as demonstrated by the induction of cleavage of the poly (that is, ADP-ribose) polymerase. Additionally, tumor growth was suppressed in cells expressingMALas compared with cells not expressingMAL.

Conclusion

Our data suggest that the epigenetic inactivation ofMAL, as a candidate tumor suppressor gene, can contribute to human epithelial cell carcinoma and may be served as a biomarker in HNSCC.
Role of the atypical chemoattractant receptor CRAM in regulating CCL19 induced CCR7 responses in B-cell chronic lymphocytic leukemia
Authors: Catusse Julie , Leick Marion , Groch Mareike , Clark David J , Buchner Maike V , Zirlik Katja , Burger Meike
Published: November 2010

Background

The non-signalling chemokine receptors, including receptors DARC, D6 and CCX-CKR, have recently been shown to be involved in chemokine clearance and activity regulation. The human chemokine receptor CRAM (also known as HCR or CCRL2) is the most recently identified member of this atypical group. CRAM is expressed on B cells in a maturation-stage dependent manner and absent on T cells. We have recently shown that it competitively binds CCL19. CCL19 and its signalling receptor CCR7 are critical components involved in cell recruitment to secondary lymphoid organs and in maturation. B cell Chronic Lymphocytic Leukemia (B-CLL) is a low-grade lymphoma characterized by proliferative centres (or pseudofollicles). Proliferative centres develop due to abnormal cellular localisation and they are involved in the development of malignant cells. CCR7 is highly expressed on B cells from CLL patients and mediates migration towards its ligands CCL19 and CCL21, while CRAM expression and potential interferences with CCR7 are yet to be characterized.

Results

In this study, we show that B cells from patients with B-CLL present highly variable degrees of CRAM expression in contrast to more consistently high levels of CCR7. We investigated the hypothesis that, similar to the atypical receptor DARC, CRAM can modulate chemokine availability and/or efficacy, resulting in the regulation of cellular activation. We found that a high level of CRAM expression was detrimental to efficient chemotaxis with CCL19. MAP-kinase phosphorylation and intracellular calcium release induced by CCL19 were also altered by CRAM expression. In addition, we demonstrate that CRAM-induced regulation of CCL19 signalling is maintained over time.

Conclusions

We postulate that CRAM is a factor involved in the fine tuning/control of CCR7/CCL19 mediated responses. This regulation could be critical to the pivotal role of CCL19 induced formation of proliferation centres supporting the T/B cells encounter as well as disease progression in B-CLL.
Effects of Kurozu concentrated liquid on adipocyte size in rats
Authors: Tong Li-Tao , Katakura Yoshinori , Kawamura Sayaka , Baba Sanae , Tanaka Yasutake , Udono Miyako , Kondo Yoshie , Nakamura Kumi , Imaizumi Katsumi , Sato Masao
Published: November 2010

Background

Kurozu concentrated liquid (KCL) is used as a health-promoting supplement for the treatment of disorders such as cancer, hyperlipidemia, and hypertension in Japan. We investigated the possible anti-obesity effects of KCL in rats.

Methods

Male Sprague Dawley rats were fed American Institute of Nutrition 76 formula diet and were orally administrated KCL or acetic acid at a dose of 100 mg/kg body weight or deionized water for 4 weeks. Adipocyte size, DNA content in subcutaneous adipose tissue, lipid levels in the serum and liver, and the rate of fatty acid excretion were determined. Effects of KCL on pancreatic lipase activity and 3T3-L1 preadipocyte differentiation were investigated in vitro.

Results

In the KCL group, the average adipocyte size in subcutaneous and perirenal adipose tissues was significantly reduced. The KCL-administered rats displayed greater numbers of small adipocytes in the subcutaneous, perirenal and mesenteric adipose tissues than did rats from the other groups. In the KCL group, the DNA content in subcutaneous adipose tissue was significantly increased. The rate of fatty acid excretion was significantly increased in the KCL group. Furthermore, KCL significantly inhibited pancreatic lipase activity in vitro, and also significantly inhibited fat accumulation and mRNA expression of fatty acid binding protein 2 (aP2) and peroxisome proliferator-activated γ (PPARγ) in 3T3-L1 preadipocyte. The levels of serum and liver lipids, the concentration of serum glucose, and the levels of adiponectin were similar among the 3 groups.

Conclusion

Oral administration of KCL decreases the adipocyte size via inhibition of dietary fat absorption and reductions of PPARγ and aP2 mRNA expression levels in adipocytes.
Detection of a novel porcine parvovirus, PPV4, in chinese swine herds
Authors: Huang Lv , Zhai Shao-Lun , Cheung Andrew K , Zhang Hong-Biao , Long Jin-Xue , Yuan Shi-Shan
Published: November 2010

To determine whether the novel porcine parvovirus type 4 (PPV4) recently reported in America is prevalent in China, a set of specific primers was designed and used for molecular survey of PPV4 among the clinical samples collected from various provinces of China between 2006 and 2010. The results showed that PPV4 is present in Chinese swine herds at a rate of 2.09% (12/573) among the clinical samples examined and 0.76% (1/132) among the samples taken from healthy animals. We also noted that PPV4 was not detected in samples taken prior to 2009. Analysis of the coding sequences showed that the Chinese and American PPV4 genome sequences are closely related with greater than 99% nucleotide sequence identity. Similar to a previous study, viral genomes in head-to-tail configuration of various lengths of the non-coding region were detected. Our findings confirmed that PPV4 is a unique recently discovered virus in pigs. Phylogenetically, PPV4 is most closely related to bovine parvovirus 2 (BPV2, which is not a Bocavirus and is not assigned to any Parvovirinae genus) and shares limited ORF1 (33.6%) and ORF2 (24.5%) amino acid identity. With respect to genome structure and organization, PPV4 encodes an ORF3 in the middle of the viral genome that resembles the Bocavirus genus. However, the PPV4 ORF3 encoded protein shares minimal amino acid identity with the ORF3 encoded proteins of the Bocavirus genus.
Prevalence of active hepatitis c virus infection in district mansehra pakistan
Authors: Ali Amjad , Ahmad Habib , Ali Ijaz , Khan Sheema , Zaidi Gulshan , Idrees Muhammad
Published: November 2010

Prevalence of active hepatitis C virus (HCV) infection in apparently healthy inhabitants of District Mansehra, Pakistan was surveyed during September, 2009 to May, 2010. Subjects of different age and gender groups were analyzed through random blood sampling from people of three areas viz; Tehsil Mansehra, Tehsil Balakot and Tehsil Oghi. Sum of 400 individuals, 300 male and 100 females with age groups from 10 years to 50 and above were included in the study. All the individuals were screened for antibodies against HCV. The positive samples thus screened, were subjected to polymerase chain reaction (PCR) analysis for detection of HCV-RNA. The results showed that 3.5% of the people of District Mansehra are actively infected with HCV whereas 7% of the population in general, has the presence of antibodies against HCV in their blood. It was also concluded that the prevalence of active HCV infection was high 4% in males as compared to females (2%). The prevalence of HCV proportionality increases with the increase in age of the people. Its incidence was highest (7.69%) in the people of the age group of 51 years and above, whereas no sign of infection was recorded for the age group of 10-20 years.
Increased expression of sialic acid in cervical biopsies with squamous intraepithelial lesions
Authors: López-Morales Dolores , Reyes-Leyva Julio , Santos-López Gerardo , Zenteno Edgar , Vallejo-Ruiz Verónica
Published: November 2010

Background

Altered sialylation has been observed during oncogenic transformation. Sialylated oligosaccharides of glycoproteins and glycolipids have been implicated in tumor progression and metastases. In the cervical cancer high levels of sialic acid have been reported in the patients serum, and an increased of total sialic acid concentration has been reported for the cervical neoplasia and cervical cancer. This study investigates the changes in expression and distribution of α2,3-linked sialic acid and α2,6- linked sialic acid in low and high squamous intraepithelial lesions and in normal tissue.

Methods

Lectin histochemistry was used to examine the expression and distribution of sialic acid in different grades of cervical neoplasia. We applied Maackia amurensis lectin, which interacts with α2,3-linked sialic acid and Sambucus nigra lectin specific for α2,6-linked sialic acid.

Results

The histochemical analysis showed that α2,3-linked sialic acid and α2,6- linked sialic acid increased in intensity and distribution in concordance with the grade of squamous intraepithelial lesion (SIL). These results are in concordance with a previous study that reports increased RNAm levels of three sialyltransferases.

Conclusions

These results show that the change in sialylation occurs before cancer development and may play an important role in cellular transformation. These findings provide the basis for more detailed studies of the possible role of cell surface glycoconjugates bearing sialic acid in the cellular cervix transformation.
A biologically competitive 21 days hypofractionation scheme with weekly concomitant boost in breast cancer radiotherapy feasibility acute sub-acute and short term late effects
Authors: Guenzi Marina , Vagge Stefano , Azinwi Ngwa Che , D'Alonzo Alessia , Belgioia Liliana , Garelli Stefania , Gusinu Marco , Corvò Renzo
Published: November 2010

Background

Radiation therapy after lumpectomy is a standard part of breast conserving therapy for invasive breast carcinoma. The most frequently used schedule worldwide is 60 Gy in 30 fractions in 6 weeks, a time commitment that sporadically may dissuade some otherwise eligible women from undertaking treatment. The purpose and primary endpoint of this perspective study is to evaluate feasibility and short-term late toxicity in a hypofractionated whole breast irradiation schedule.

Methods

Between February and October 2008 we treated 65 consecutive patients with operable invasive early-stage breast cancer with a hypofractionated schedule of external beam radiation therapy. All patients were assigned to 39 Gy in 13 fractions in 3 weeks to the whole breast plus a concomitant weekly boost dose to the lumpectomy cavity of 3 Gy in 3 fractions.

Results

All the patients had achieved a median follow up of 24 months (range 21-29 months). At the end of treatment 52% presented grade 0 acute toxicity 39% had grade 1 and 9% had grade 2. At 6 months with all the patients assessed there were 34% case of grade 1 subacute toxicity and 6% of grade 2. At 12 months 43% and 3% of patients presented with clinical grade 1 and grade 2 fibrosis respectively and 5% presented grade 1 hyperpigmentation. The remaining patients were free of side effects. At 24 months, with 56 assessed, just 2 patients (3%) showed grade 2 of late fibrosis.

Conclusions

The clinical results observed showed a reasonably good feasibility of the accelerated hypofractionated schedule in terms of acute, subacute and short-term late toxicity. This useful 13 fractions with a concomitant boost schedule seems, in selected patients, a biologically acceptable alternative to the traditional 30 days regime.
Development and analysis of an in vivo-compatible metabolic network of Mycobacterium tuberculosis
Authors: Fang Xin , Wallqvist Anders , Reifman Jaques
Published: November 2010

Background

During infection, Mycobacterium tuberculosis confronts a generally hostile and nutrient-poor in vivo host environment. Existing models and analyses of M. tuberculosis metabolic networks are able to reproduce experimentally measured cellular growth rates and identify genes required for growth in a range of different in vitro media. However, these models, under in vitro conditions, do not provide an adequate description of the metabolic processes required by the pathogen to infect and persist in a host.

Results

To better account for the metabolic activity of M. tuberculosis in the host environment, we developed a set of procedures to systematically modify an existing in vitro metabolic network by enhancing the agreement between calculated and in vivo- measured gene essentiality data. After our modifications, the new in vivo network contained 663 genes, 838 metabolites, and 1,049 reactions and had a significantly increased sensitivity (0.81) in predicted gene essentiality than the in vitro network (0.31). We verified the modifications generated from the purely computational analysis through a review of the literature and found, for example, that, as the analysis suggested, lipids are used as the main source for carbon metabolism and oxygen must be available for the pathogen under in vivo conditions. Moreover, we used the developed in vivo network to predict the effects of double-gene deletions on M. tuberculosis growth in the host environment, explore metabolic adaptations to life in an acidic environment, highlight the importance of different enzymes in the tricarboxylic acid-cycle under different limiting nutrient conditions, investigate the effects of inhibiting multiple reactions, and look at the importance of both aerobic and anaerobic cellular respiration during infection.

Conclusions

The network modifications we implemented suggest a distinctive set of metabolic conditions and requirements faced by M. tuberculosis during host infection compared with in vitro growth. Likewise, the double-gene deletion calculations highlight the importance of specific metabolic pathways used by the pathogen in the host environment. The newly constructed network provides a quantitative model to study the metabolism and associated drug targets of M. tuberculosis under in vivo conditions.
Is advanced life support better than basic life support in prehospital care? A systematic review
Authors: Ryynänen Olli-Pekka , Iirola Timo , Reitala Janne , Pälve Heikki , Malmivaara Antti
Published: November 2010

Background -

Prehospital care is classified into ALS- (advanced life support) and BLS- (basic life support) levels according to the methods used. ALS-level prehospital care uses invasive methods, such as intravenous fluids, medications and intubation. However, the effectiveness of ALS care compared to BLS has been questionable.

Aim -

The aim of this systematic review is to compare the effectiveness of ALS- and BLS-level prehospital care.

Material and methods -

In a systematic review, articles where ALS-level prehospital care was compared to BLS-level or any other treatment were included. The outcome variables were mortality or patient's health-related quality of life or patient's capacity to perform daily activities.

Results -

We identified 46 articles, mostly retrospective observational studies. The results on the effectiveness of ALS in unselected patient cohorts are contradictory. In cardiac arrest, early cardiopulmonary resuscitation and defibrillation are essential for survival, but prehospital ALS interventions have not improved survival. Prehospital thrombolytic treatment reduces mortality in patients having a myocardial infarction. The majority of research into trauma favours BLS in the case of penetrating trauma and also in cases of short distance to a hospital. In patients with severe head injuries, ALS provided by paramedics and intubation without anaesthesia can even be harmful. If the prehospital care is provided by an experienced physician and by a HEMS organisation (Helicopter Emergency Medical Service), ALS interventions may be beneficial for patients with multiple injuries and severe brain injuries. However, the results are contradictory.

Conclusions -

ALS seems to improve survival in patients with myocardial infarction and BLS seems to be the proper level of care for patients with penetrating injuries. Some studies indicate a beneficial effect of ALS among patients with blunt head injuries or multiple injuries. There is also some evidence in favour of ALS among patients with epileptic seizures as well as those with a respiratory distress.
Hepatitis C new drugs
Authors: Wedemeyer H
Published: November 2010

7‐11 November 2010, Tenth International Congress on Drug Therapy in HIV Infection, Glasgow, UK
Highlights of the 12th International Workshop on Adverse Drug Reactions and Co‐Morbidities in HIV, London, November 2010
Authors: Schambelan M
Published: November 2010

7‐11 November 2010, Tenth International Congress on Drug Therapy in HIV Infection, Glasgow, UK
Offering HIV testing in an emergency admission unit in Newcastle upon Tyne, UK — a pilot audit study
Authors: Ellis S , Graham L , Price DA , Ong ELC
Published: November 2010

7‐11 November 2010, Tenth International Congress on Drug Therapy in HIV Infection, Glasgow, UK
Correlates of hepatic stiffness by FibroScan© in a multicentric Italian cohort of HIV‐infected patients
Authors: Tarquini P , Sozio F , Marchionni E , Gabrielli E , Polilli E , Ancarani F , Di Giammartino D , Manzoli LM , Parruti G
Published: November 2010

7‐11 November 2010, Tenth International Congress on Drug Therapy in HIV Infection, Glasgow, UK
Kidney tubular function and serum phosphate levels in HIV‐1‐infected patients treated with tenofovir: preliminary results
Authors: Grima P , Zizza A , Calabrese P , Chiavaroli R , Faraone A , Tana M , Sodo MR , Tundo P , Guido M
Published: November 2010

7‐11 November 2010, Tenth International Congress on Drug Therapy in HIV Infection, Glasgow, UK
Proteomic approach with LCMS-IT-TOF identified an increase of Rab33B after transient focal cerebral ischemia in mice
Authors: Hyakkoku Kana , Hamanaka Junya , Tsuruma Kazuhiro , Shimazawa Masamitsu , Hara Hideaki
Published: November 2010

Background

Several proteins are known to be markedly expressed in the brain during cerebral ischemia; however, the changes in protein profiles within the ischemic brain after an ischemic insult have not been fully elucidated. We studied the changes in the ischemic brain proteome after focal cerebral ischemia, induced by middle cerebral artery occlusion (MCAO) in mice.

Methods

LCMS-IT-TOF mass spectrometry was used to detect the changes in ischemic brain protein patterns after MCAO. We evaluated the protein expression detected in the ischemic area, by western blotting and immunohistochemistry.

Results

Nine unique proteins were identified from the ischemic area at 10 h after ischemic insult. Among these proteins, we focused on Rab33b, a member of RAS oncogene family and we found that Rab33b was up-regulated in the ischemic striatum and the number of Rab33B-positive cells increased in a time-dependent manner. Rab33B colocalized with Iba-1 positive microglia in the ischemic area.

Conclusion

These findings suggest that LCMS-IT-TOF is useful for identifying changes in proteins after cerebral ischemia and that Rab33B is partially related to the pathogenesis of transient cerebral ischemia in mice.
Evidence-Based Guidelines for Determination of Sample Size and Interpretation of the European Organisation for the Research and Treatment of Cancer Quality of Life Questionnaire Core 30
Authors: Cocks Kim , King Madeleine T. , Velikova Galina , Martyn St-James Marrissa , Fayers Peter M. , Brown Julia M.
Published: November 2010

Purpose

To use published literature to estimate large, medium, and small differences in quality of life (QOL) data from the European Organisation for the Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30).

Methods

An innovative method combining systematic review of published studies, expert opinions, and meta-analysis was used to estimate large, medium, and small differences for QLQ-C30 scores. Published mean data were identified from the literature. Differences (contrasts) between groups (eg, between treatment groups, age groups, and performance status groups) were reviewed by 34 experts in QOL measurement and cancer treatment. The experts, blinded to actual QOL results, were asked to predict these differences. A large difference was defined as one representing unequivocal clinical relevance. A medium difference was defined as likely to be clinically relevant but to a lesser extent. A small difference was one believed to be subtle but nevertheless clinically relevant. A trivial difference was used to describe circumstances unlikely to have any clinical relevance. Actual QOL results were combined using meta-analytic techniques to estimate differences corresponding to small, medium, or large effects.

Results

Nine hundred eleven articles were identified, leading to 152 relevant articles (2,217 contrasts) being reviewed by at least two experts. Resulting estimates from the meta-analysis varied depending on the subscale. Thus, the recommended minimum to detect medium differences ranges from 9 (cognitive functioning) to 19 points (role functioning).

Conclusion

Guidelines for the size of effects are provided for the QLQ-C30 subscales. These guidelines can be used for sample size calculations for clinical trials and can also be used to aid interpretation of differences in QLQ-C30 scores.
What Oncologists Believe They Said and What Patients Believe They Heard: An Analysis of Phase I Trial Discussions
Authors: Jenkins Valerie , Solis-Trapala Ivonne , Langridge Carolyn , Catt Susan , Talbot Denis C. , Fallowfield Lesley J.
Published: November 2010

Purpose

Evaluation of the communication and informed consent process in phase I clinical trial interviews to provide authentic, practice-based content for inclusion in a communication skills training intervention for health care professionals.

Patients and Methods

Seventeen oncologists and 52 patients from five United Kingdom cancer centers consented to recording of phase I trial discussions. Following each consultation, clinicians completed questionnaires indicating areas they felt they had discussed, and researchers conducted semistructured interviews with patients examining their recall and understanding. Patients and oncologists also completed the Life Orientation Test-Revised questionnaire, measuring predisposition toward optimism. Independent researchers coded the consultations identifying discussion of key information areas and how well this was done. Observed levels of agreement were analyzed for each consultation between oncologist-coder, oncologist-patient, and patient-coder pairs.

Results

In several key areas, information was either missing or had been explained but was interpreted incorrectly by patients. Discussion of prognosis was a frequent omission, with patients and coders significantly more likely to agree that oncologists had not discussed it (odds, 4.8; P < .001). In contrast, coders and oncologists were more likely to agree that alternate care plans to phase I trial entry had been explained (odds, 2.5; P = .023).

Conclusion

These data indicate that fundamental components of communication and information sharing about phase I trial participation are often missing from interviews. Important omissions included discussion of prognosis and ensuring patient understanding about supportive care. These findings will inform educational initiatives to assist communication about phase I trials.
Gene Expression Signature to Improve Prognosis Prediction of Stage II and III Colorectal Cancer
Authors: Salazar Ramon , Roepman Paul , Capella Gabriel , Moreno Victor , Simon Iris , Dreezen Christa , Lopez-Doriga Adriana , Santos Cristina , Marijnen Corrie , Westerga Johan , Bruin Sjoerd , Kerr David , Kuppen Peter , van de Velde Cornelis , Morreau Hans , Van Velthuysen Loes , Glas Annuska M. , Van't Veer Laura J. , Tollenaar Rob
Published: November 2010

Purpose

This study aims to develop a robust gene expression classifier that can predict disease relapse in patients with early-stage colorectal cancer (CRC).

Patients and Methods

Fresh frozen tumor tissue from 188 patients with stage I to IV CRC undergoing surgery was analyzed using Agilent 44K oligonucleotide arrays. Median follow-up time was 65.1 months, and the majority of patients (83.6%) did not receive adjuvant chemotherapy. A nearest mean classifier was developed using a cross-validation procedure to score all genes for their association with 5-year distant metastasis–free survival.

Results

An optimal set of 18 genes was identified and used to construct a prognostic classifier (ColoPrint). The signature was validated on an independent set of 206 samples from patients with stage I, II, and III CRC. The signature classified 60% of patients as low risk and 40% as high risk. Five-year relapse-free survival rates were 87.6% (95% CI, 81.5% to 93.7%) and 67.2% (95% CI, 55.4% to 79.0%) for low- and high-risk patients, respectively, with a hazard ratio (HR) of 2.5 (95% CI, 1.33 to 4.73; P = .005). In multivariate analysis, the signature remained one of the most significant prognostic factors, with an HR of 2.69 (95% CI, 1.41 to 5.14; P = .003). In patients with stage II CRC, the signature had an HR of 3.34 (P = .017) and was superior to American Society of Clinical Oncology criteria in assessing the risk of cancer recurrence without prescreening for microsatellite instability (MSI).

Conclusion

ColoPrint significantly improves the prognostic accuracy of pathologic factors and MSI in patients with stage II and III CRC and facilitates the identification of patients with stage II disease who may be safely managed without chemotherapy.
Disease Course Patterns After Discontinuation of Bevacizumab: Pooled Analysis of Randomized Phase III Trials
Authors: Miles David , Harbeck Nadia , Escudier Bernard , Hurwitz Herbert , Saltz Leonard , Van Cutsem Eric , Cassidy Jim , Mueller Barbara , Sirzén Florin
Published: November 2010

Purpose

Preclinical studies have suggested accelerated tumor growth, local invasion, and distant metastasis after withdrawal of treatment with some antiangiogenic agents. To investigate whether discontinuation of bevacizumab treatment is associated with accelerated disease progression or increased mortality, we retrospectively analyzed five randomized, placebo-controlled phase III studies in 4,205 patients with breast, colorectal, renal, and pancreatic cancer.

Methods

Time from treatment discontinuation to progressive disease or death was analyzed in patients discontinuing bevacizumab/placebo as a result of adverse events (AEs). Mortality rates were assessed at 30, 60, 90, 120, 150, 180, and 210 days after the last bevacizumab/placebo dose in the following two groups: patients discontinuing bevacizumab/placebo as a result of AEs and patients discontinuing bevacizumab/placebo for any reason. In the same groups, time from treatment discontinuation to death was analyzed. Data on disease progression pattern were available and analyzed in four of the five studies.

Results

In the pooled analysis, median time from discontinuation as a result of AEs to progression/death was 3.0 months (95% CI, 2.6 to 3.8 months) for placebo and 4.0 months (95% CI, 3.4 to 4.6 months) for bevacizumab (hazard ratio, 0.93; 95% CI, 0.79 to 1.10). Mortality rates from 30 days to 210 days after treatment discontinuation and time from discontinuation to death were similar in bevacizumab- and placebo-treated patients. In addition, similar patterns of disease progression were seen in bevacizumab- and placebo-treated patients.

Conclusion

This retrospective analysis of five placebo-controlled clinical trials does not support a decreased time to disease progression, increased mortality, or altered disease progression pattern after cessation of bevacizumab therapy.
The neural crest-enriched microRNA miR-452 regulates epithelial-mesenchymal signaling in the first pharyngeal arch
Authors: Sheehy Neil T. , Cordes Kimberly R. , White Mark P. , Ivey Kathryn N. , Srivastava Deepak
Published: November 2010

Neural crest cells (NCCs) are a subset of multipotent, migratory stem cells that populate a large number of tissues during development and are important for craniofacial and cardiac morphogenesis. Although microRNAs (miRNAs) have emerged as important regulators of development and disease, little is known about their role in NCC development. Here, we show that loss of miRNA biogenesis by NCC-specific disruption of murine Dicer results in embryos lacking craniofacial cartilaginous structures, cardiac outflow tract septation and thymic and dorsal root ganglia development. Dicer mutant embryos had reduced expression of Dlx2, a transcriptional regulator of pharyngeal arch development, in the first pharyngeal arch (PA1). miR-452 was enriched in NCCs, was sufficient to rescue Dlx2 expression in Dicer mutant pharyngeal arches, and regulated non-cell-autonomous signaling involving Wnt5a, Shh and Fgf8 that converged on Dlx2 regulation in PA1. Correspondingly, knockdown of miR-452 in vivo decreased Dlx2 expression in the mandibular component of PA1, leading to craniofacial defects. These results suggest that post-transcriptional regulation by miRNAs is required for differentiation of NCC-derived tissues and that miR-452 is involved in epithelial-mesenchymal signaling in the pharyngeal arch.
A functionally conserved boundary element from the mouse HoxD locus requires GAGA factor inDrosophila
Authors: Vasanthi Dasari , Anant Mamidi , Srivastava Surabhi , Mishra Rakesh K.
Published: November 2010

Hox genes are necessary for proper morphogenesis and organization of various body structures along the anterior-posterior body axis. These genes exist in clusters and their expression pattern follows spatial and temporal co-linearity with respect to their genomic organization. This colinearity is conserved during evolution and is thought to be constrained by the regulatory mechanisms that involve higher order chromatin structure. Earlier studies, primarily in Drosophila, have illustrated the role of chromatin-mediated regulatory processes, which include chromatin domain boundaries that separate the domains of distinct regulatory features. In the mouse HoxD complex, Evx2 and Hoxd13 are located ∼9 kb apart but have clearly distinguishable temporal and spatial expression patterns. Here, we report the characterization of a chromatin domain boundary element from the Evx2-Hoxd13 region that functions in Drosophila as well as in mammalian cells. We show that the Evx2-Hoxd13 region has sequences conserved across vertebrate species including a GA repeat motif and that the Evx2-Hoxd13 boundary activity in Drosophila is dependent on GAGA factor that binds to the GA repeat motif. These results show that Hox genes are regulated by chromatin mediated mechanisms and highlight the early origin and functional conservation of such chromatin elements.
Cellular stress induces cytoplasmic RNA granules in fission yeast
Authors: Nilsson Daniel , Sunnerhagen Per
Published: November 2010

Severe stress causes plant and animal cells to form large cytoplasmic granules containing RNA and proteins. Here, we demonstrate the existence of stress-induced cytoplasmic RNA granules inSchizosaccharomyces pombe. Homologs to several known protein components of mammalian processing bodies and stress granules are found in fission yeast RNA granules. In contrast to mammalian cells, poly(A)-binding protein (Pabp) colocalizes in stress-induced granules with decapping protein. After glucose deprivation, protein kinase A (PKA) is required for accumulation of Pabp-positive granules and translational down-regulation. This is the first demonstration of a role for PKA in RNA granule formation. In mammals, the translation initiation protein eIF2α is a key regulator of formation of granules containing poly(A)-binding protein. InS. pombe, nonphosphorylatable eIF2α does not block but delays granule formation and subsequent clearance after exposure to hyperosmosis. At least two separate pathways inS. pombeappear to regulate stress-induced granules:pka1mutants are fully proficient to form granules after hyperosmotic shock; conversely, eIF2α does not affect granule formation in glucose starvation. Further, we demonstrate a Pka1-dependent link between calcium perturbation and RNA granules, which has not been described earlier in any organism.
Quantitative approaches to monitor protein–nucleic acid interactions using fluorescent probes
Authors: Pagano John M. , Clingman Carina C. , Ryder Sean P.
Published: November 2010

Sequence-specific recognition of nucleic acids by proteins is required for nearly every aspect of gene expression. Quantitative binding experiments are a useful tool to measure the ability of a protein to distinguish between multiple sequences. Here, we describe the use of fluorophore-labeled oligonucleotide probes to quantitatively monitor protein/nucleic acid interactions. We review two complementary experimental methods, fluorescence polarization and fluorescence electrophoretic mobility shift assays, that enable the quantitative measurement of binding affinity. We also present two strategies for post-synthetic end-labeling of DNA or RNA oligonucleotides with fluorescent dyes. The approaches discussed here are efficient and sensitive, providing a safe and accessible alternative to the more commonly used radio-isotopic methods.
Neptunium(V) complexation by natural pyoverdins and related model compounds
Authors: Moll Henry , Glorius Maja , Johnsson A. , Schäfer M. , Budzikiewicz H. , Pedersen K. , Bernhard Gert
Published: November 2010

Ubiquitous fluorescentPseudomonasspecies secrete bacterial pyoverdin-type siderophores. These bioligands have great potential to bind and transport actinides in the environment due to their hydroxamate and catechol functionalities. We investigated the unknown interaction of the neptunyl cation (NpO2+) with pyoverdins (PYO) released byPseudomonas fluorescens(CCUG 32456) cells and with simple hydroxamate (salicylhydroxamic acid: SHA and benzohydroxamic acid: BHA) and catechol (2,3-dihydroxynaphthalene: NAP) ligands using near-infrared (NIR) absorption spectroscopy over a wide pH range. NpO2+-bioligand species of the MxLyHztype were identified from the spectrophotometric titrations in all four systems. The 1:1:2, 1:1:1, and 1:1:0 complexes were determined with the pyoverdins. In addition to 1:1 species, SHA, BHA, and NAP also form 1:2:0 species with NpO2+. The stability constants of these neptunyl(V)-bioligand complexes and their individual spectroscopic properties are reported. Our findings indicate that NpO2+has a stronger affinity to the catechol functionality of the pyoverdin molecule. The identified NpO2+-PYO species belong to the strongest NpO2+complexes with organic material reported so far.
Influence of Boom Clay organic matter on the adsorption of Eu3+by illite – geochemical modelling using the component additivity approach
Authors: Bruggeman C. , Liu D. J. , Maes Norbert
Published: November 2010

The solid–liquid distribution of europium (Eu) between an adsorptive surface and a solution phase containing a competitive colloid is the result of a delicate balance between several individual chemical reactions. In this study, adsorption isotherms of Eu in presence of dissolved Boom Clay natural organic matter were experimentally determined under conditions relevant for a geological repository (trace Eu concentrations, anoxic conditions, ∼0.014 mol l−1NaHCO3background electrolyte). It was found that both the concentration and size distribution (or operational cut-off used to discriminate between “mobile” and “immobile” colloids) of natural organic matter has a strong influence on the observed solid–liquid distribution.

The experimental data were subsequently modelled using a component additive approach with two well-established sorption/interaction models: the 2 SPNE SC/CE model for describing Eu adsorption on illite, and Humic Ion-Binding Model VI for describing Eu complexation to natural organic matter. Model parameters were gathered from dedicated measurements in batch systems containing only Eu and the interacting phase under study, under similar conditions as in the ternary isotherm experiments. Mutual interactions between illite and natural organic matter were studied and quantified. Under the experimental conditions of this study, it was found that these interactions were only of minor importance.

The two models were subsequently combined to blind predict the Eu solid–liquid distribution in the ternary batch experiments. Within an error margin of 0.5logߙKdunits, the additivity approach succeeded well in predicting Eu uptake in all experimental systems studied. A sensitivity analysis was performed to select the most important model parameters influencing the Eu uptake, and the robustness of the model. This study has shown that the component additivity approach for describing and predicting uptake of trivalent lanthanides/actinides under Boom Clay conditions, is promising, and may help in unraveling the complex behaviour of these radionuclides witnessed in migration experiments.
The role of green rust in the migration of radionuclides: An overview of processes that can control mobility of radioactive elements in the environment using as examples Np, Se and Cr
Authors: Skovbjerg L. L. , Christiansen B. C. , Nedel S. , Dideriksen K. , Stipp S. L. S.
Published: November 2010

Green rust is an Fe(II)-Fe(III) layered double hydroxide that can be present in groundwater in rock formations where long term nuclear waste repositories are being planned. Green rust (GR) is redox-active, making data for its behaviour a necessity in safety assessment modeling because the mobility of many elements, including radionuclides, is controlled by redox reactions. In this paper, we compare the influence of geometry and charge of three redox active ions on their reaction with colloidal size particles of green rust sodium sulfate (GRNa,SO4). Pentavalent neptunyl, selenite and chromate all have different geometries and valence states. Ions with shape and charge similar to SO4
2−, the GRNa,SO4 interlayer anion such as, for example, CrO4
2−, diffuse into the GR interlayer where electrons are readily transferred, and the element may become incorporated into the final oxidised product. More often, the geometry and/or charge of the ion differ significantly from the interlayer anion, such as is the case for SeO3
2− and NpO2
+. These are reduced at the exterior of the flat hexagonal GR crystals, primarily at the edges. Se(IV) is reduced to Se(0) and precipitates as a separate phase, while Np(V) is reduced to Np(IV) and precipitates possibly as NpO2 or as a substitute in an iron oxide. Whether it adsorbs to pore walls in the groundwater flowpath, or it travels as a colloid, green rust can have a significant effect on radionuclide mobility in the near and far field of repositories.