bluuebunny/crossref_metadata_2025_split huggingface dataset:
Heterologous expression of glucose oxidase in the yeast Kluyveromyces marxianus
Authors: Rocha Saul N , Abrahão-Neto José , Cerdán María E , González-Siso María I , Gombert Andreas K 
Published: January 2010

Abstract

Background
In spite of its advantageous physiological properties for bioprocess applications, the use of the yeast Kluyveromyces marxianus as a host for heterologous protein production has been very limited, in constrast to its close relative Kluyveromyces
lactis. In the present work, the model protein glucose oxidase (GOX) from Aspergillus niger was cloned into K. marxianus CBS 6556 and into K. lactis CBS 2359 using three different expression systems. We aimed at verifying how each expression system would affect protein expression, secretion/localization, post-translational modification, and biochemical properties.

Results
The highest GOX expression levels (1552 units of secreted protein per gram dry cell weight) were achieved using an episomal system, in which the INU1 promoter and terminator were used to drive heterologous gene expression, together with the INU1 prepro sequence, which was employed to drive secretion of the enzyme. In all cases, GOX was mainly secreted, remaining either in the periplasmic space or in the culture supernatant. Whereas the use of genetic elements from Saccharomyces cerevisiae to drive heterologous protein expression led to higher expression levels in K. lactis than in K. marxianus, the use of INU1 genetic elements clearly led to the opposite result. The biochemical characterization of GOX confirmed the correct expression of the protein and showed that K. marxianus has a tendency to hyperglycosylate the protein, in a similar way as already observed for other yeasts, although this tendency seems to be smaller than the one of e.g. K. lactis and S. cerevisiae. Hyperglycosylation of GOX does not seem to affect its affinity for the substrate, nor its activity.

Conclusions
Taken together, our results indicate that K. marxianus is indeed a good host for the expression of heterologous proteins, not only for its physiological properties, but also because it correctly secretes and folds these proteins.
Isolation, characterization and heterologous expression of a novel chitosanase from Janthinobacterium sp. strain 4239
Authors: Johnsen Mads G , Hansen Ole C , Stougaard Peter 
Published: January 2010

Abstract

Background
Chitosanases (EC 3.2.1.132) hydrolyze the polysaccharide chitosan, which is composed of partially acetylated β-(1,4)-linked glucosamine residues. In nature, chitosanases are produced by a number of Gram-positive and Gram-negative bacteria, as well as by fungi, probably with the primary role of degrading chitosan from fungal and yeast cell walls for carbon metabolism. Chitosanases may also be utilized in eukaryotic cell manipulation for intracellular delivery of molecules formulated with chitosan as well as for transformation of filamentous fungi by temporal modification of the cell wall structures.
However, the chitosanases used so far in transformation and transfection experiments show optimal activity at high temperature, which is incompatible with most transfection and transformation protocols. Thus, there is a need for chitosanases, which display activity at lower temperatures.

Results
This paper describes the isolation of a chitosanase-producing, cold-active bacterium affiliated to the genus Janthinobacterium. The 876 bp chitosanase gene from the Janthinobacterium strain was isolated and characterized. The chitosanase was related to the Glycosyl Hydrolase family 46 chitosanases with Streptomyces chitosanase as the closest related (64% amino acid sequence identity). The chitosanase was expressed recombinantly as a periplasmic enzyme in Escherichia coli in amounts about 500 fold greater than in the native Janthinobacterium strain. Determination of temperature and pH optimum showed that the native and the recombinant chitosanase have maximal activity at pH 5-7 and at 45°C, but with 30-70% of the maximum activity at 10°C and 30°C, respectively.

Conclusions
A novel chitosanase enzyme and its corresponding gene was isolated from Janthinobacterium and produced recombinantly in E. coli as a periplasmic enzyme. The Janthinobacterium chitosanase displayed reasonable activity at 10°C to 30°C, temperatures that are preferred in transfection and transformation experiments.
Role of senescence and mitotic catastrophe in cancer therapy
Authors: Singh Richa , George Jasmine , Shukla Yogeshwer 
Published: January 2010

Abstract
Senescence and mitotic catastrophe (MC) are two distinct crucial non-apoptotic mechanisms, often triggered in cancer cells and tissues in response to anti-cancer drugs. Chemotherapeuticals and myriad other factors induce cell eradication via these routes. While senescence drives the cells to a state of quiescence, MC drives the cells towards death during the course of mitosis. The senescent phenotype distinguishes tumor cells that survived drug exposure but lost the ability to form colonies from those that recover and proliferate after treatment. Although senescent cells do not proliferate, they are metabolically active and may secrete proteins with potential tumor-promoting activities. The other anti-proliferative response of tumor cells is MC that is a form of cell death that results from abnormal mitosis and leads to the formation of interphase cells with multiple micronuclei. Different classes of cytotoxic agents induce MC, but the pathways of abnormal mitosis differ depending on the nature of the inducer and the status of cell-cycle checkpoints. In this review, we compare the two pathways and mention that they are activated to curb the growth of tumors. Altogether, we have highlighted the possibilities of the use of senescence targeting drugs, mitotic kinases and anti-mitotic agents in fabricating novel strategies in cancer control.
Comparison of the epidermal growth factor receptor protein expression between primary non-small cell lung cancer and paired lymph node metastases: implications for targeted nuclide radiotherapy
Authors: Rao Chuangzhou , Hu Qiongge , Ma Jianhua , Li Jian , Zhang Chen , Shen Li , Wei Qichun 
Published: January 2010

Abstract

Background
The knowledge of Epidermal growth factor receptor (EGFR) expression in metastases of NSCLC was limited. In receptor-mediated targeted nuclide radiotherapy, tumor cells are killed with delivered radiation and therapeutic efficiency is mainly dependent on the receptor expression. Thus, the level and stability of receptor expression in both primary tumors and corresponding metastases is crucial in the assessment of a receptor as target. The goal of this study was to evaluate whether EGFR is suitable as target for clinical therapy.

Methods
Expression of EGFR was investigated immunohistochemically in paired samples of lymph node metastases and corresponding NSCLC primary lesions (n = 51). EGFR expression was scored as 0, 1+, 2+ or 3+.

Results
Positive (1+, 2+ or 3+) EGFR immunostaining was evident in 36 of 47 (76.6%) analysed NSCLC primary tumors, and in 78.7% of the corresponding lymph node metastases. When EGFR expression is classified as positive or negative, discordance between the primary tumors and the corresponding metastases was observed in 5 cases (10.6%). EGFR overexpression (2+ or 3+) was found in 53.2% (25/47) of the NSCLC primary tumors and 59.6% of the corresponding metastases. Nine out of the 47 paired samples (19.2%) were discordant: Only three patients who had EGFR overexpression in the primary tumors showed EGFR downregulation (0 or 1+) in lymph node metastases, while six patients changed the other way around.

Conclusions
The EGFR expression in the primary tumor and the corresponding metastasis is discordant in about 10% of the patients. When overexpression is considered, the discordance is observed in about 20% of the cases. However, concerning EGFR overexpression in the primary tumors, similar expression in the metastases could be predicted with a reasonably high probability, which is encouraging for testing of EGFR targeted nuclide radiotherapy.
An incidentally detected hepatic subcapsular hematoma in a very low birth weight newborn: a case report
Authors: Ahn Hye Shin , Chang Yun-Woo , Lee Dong Whan , Kwon Kui Hyang , Yang Seung Boo 
Published: January 2010

AbstractIntroductionA hepatic subcapsular hematoma in a neonate shows a non-specific presentation such as the presence of an abdominal mass without symptoms of hemorrhage and is clinically less distinguished as compared to cases detected during an autopsy.Case presentationA neonate was delivered by vaginal delivery after 29 weeks and three days gestation with breech presentation. In a laboratory study, there were slightly increased levels of liver enzymes but the platelet count and hemoglobin level were normal. An abdomen ultrasonography and CT image demonstrated the cystic mass containing an internal thin septum with compression of the lateral margin of the right hepatic lobe and Morison's pouch. A CT image showed an irregular low-density lesion in the dome of liver that was suspected parenchymal laceration.ConclusionWe have described the sonographic and CT findings of an incidentally detected subcapsular hematoma of the liver in a neonate who showed a breech presentation, very low birth weight and was premature.
A2A adenosine receptor stimulation decreases foam cell formation by enhancing ABCA1-dependent cholesterol efflux
Authors: Bingham Taiese Crystal , Fisher Edward A , Parathath Saj , Reiss Allison B , Chan Edwin S , Cronstein Bruce N 
Published: January 2010

Abstract
Adenosine A2A receptor occupancy diminishes foam cell formation by stimulating increased reverse cholesterol transport via ABCA1.
Immune and inflammatory cells play a critical role in the pathogenesis of atherosclerotic plaques. We have demonstrated that A2ARs inhibit foam cell formation and stimulate production of ABCA1, the primary transporter of lipoproteins. We asked whether the effects of A2ARs on foam cell formation in vitro are mediated by transporters involved in reverse cholesterol transport, ABCA1 and ABCG1. Foam cells were generated from THP-1 cells by incubation with 100 nM PMA for 2 days and incubated with acLDL (50 μg/mL) plus IFN-γ (500 U/mL) ± A2AR agonist CGS-21680 (1 μM). Radiolabeled cholesterol (0.2 μCi/ml) was added to cells, and efflux was measured using a liquid scintillation counter. Lentiviral siRNA infection markedly reduces ABCA1 or ABCG1 mRNA in THP-1 cells. Despite diminished ABCG1 expression (KD), CGS-21680 inhibits foam cell formation (81+5% inhibition; P&lt;0.0001 vs. IFN-γ alone; n=3) but has no effect on foam cell formation in ABCA1 KD cells (5+3% inhibition; P&lt;0.85 vs. IFN-γ alone; n=3). The A2A agonist increases apoA-I-mediated cholesterol efflux nearly twofold in THP-1-derived macrophages (from 9.5% to 17.5+2.5% [3H]-cholesterol efflux; P&lt;0.0090 vs. control; n=3) but not in ABCA1 KD cells. Activation of Epac, a signaling molecule downstream of the A2AR, increased ABCA1 (23+5%; P&lt;0.0007 vs. control; n=3) and phospho-ABCA1 (13+5%; P&lt;0.0003 vs. control; n=3) protein. These results demonstrate that A2AR occupancy diminishes foam cell formation by stimulating increased reverse cholesterol transport via ABCA1.
Extended opportunity-based age replacement with a one-cycle criterion
Authors: Venkat D , Coolen F P A , Coolen-Schrijner P 
Published: January 2010

Age replacement is a well-established theoretical model for guidance on preventive replacement of technical units, with the intention to replace units before failure, in a manner that is most cost-effective. Traditionally, the cost criterion is based on assumed use of the same replacement strategy over a period of infinite length, enabling application of the renewal reward theorem to derive the units’ optimal preventive replacement age. However, in many situations the number of renewals involving exchangeable units is typically small, which undermines the use of this traditional cost criterion. As an alternative, minimal average costs per unit time over a single cycle has been proposed, and studied for several age replacement scenarios. In this paper, this work is briefly reviewed, followed by presentation of results on the use of the one-cycle criterion for extended opportunity-based age replacement. In this model, a unit is replaced correctively in case of failure or preventively upon reaching a specific age, as in the standard age replacement model. In addition, the unit is replaced preventively if opportunities occur during a period before the optimal age replacement time, for example in case production stops due to failure or preventive replacement of other units in the same production process. As such, costs of preventive replacement in the latter case are likely to be smaller than if preventive replacement of the specific unit requires a production stop, making it clear that it will be attractive to benefit from such opportunities occurring within a certain time range prior to scheduled preventive replacement. Optimality results for this model, with the one-cycle criterion, are presented and illustrated, and it is shown how limiting cases coincide with several other age replacement scenarios. Iskandar and Sandoh (An extended opportunity-based age replacement policy. RAIRO Ops Res., 2000, 34, 145—154) studied this model with the renewal reward criterion, but they assumed that all preventive replacements would incur the same costs. A generalization of their results is presented, allowing different costs for the two preventive replacement scenarios. This enables the comparison of the optimal replacement strategies according to the one-cycle criterion and the renewal reward criterion.
Comparison of Fluorescence In Situ Hybridization, p57 Immunostaining, Flow Cytometry, and Digital Image Analysis for Diagnosing Molar and Nonmolar Products of Conception
Authors: Kipp Benjamin R. , Ketterling Rhett P. , Oberg Trynda N. , Cousin Margot A. , Plagge Amy M. , Wiktor Anne E. , Ihrke Johnita M. , Meyers Cecelia H. , Morice William G. , Halling Kevin C. , Clayton Amy C. 
Published: January 2010

Abstract
Pathologic examination of products of conception (POC) is used to differentiate hydropic abortus (HA), partial hydatidiform mole (PM), and complete hydatidiform mole (CM). Histologic classification of POC specimens can be difficult, and ancillary testing is often required for a definitive diagnosis. This study evaluated 66 POC specimens by flow cytometry, digital image analysis, p57 immunohistochemical analysis, and fluorescence in situ hybridization (FISH). The final diagnosis, based on the combined analysis of all test results, included 33 HAs, 24 PMs, and 9 CMs. The p57 immunostain identified 9 CMs that were evaluated as nontriploid by all other techniques. FISH seems to have the best accuracy (100%) for determining whether a specimen contains a triploid chromosome complement. These data suggest that the combination of p57 and FISH seems to be the best ancillary testing strategy to aid pathologists in the appropriate identification of CM, PM, and HA in POC specimens.
Peptides Generated Ex Vivo from Serum Proteins by Tumor-Specific Exopeptidases Are Not Useful Biomarkers in Ovarian Cancer
Authors: Timms John F , Cramer Rainer , Camuzeaux Stephane , Tiss Ali , Smith Celia , Burford Brian , Nouretdinov Ilia , Devetyarov Dmitry , Gentry-Maharaj Aleksandra , Ford Jeremy , Luo Zhiyuan , Gammerman Alex , Menon Usha , Jacobs Ian 
Published: January 2010

Abstract
Background: The serum peptidome may be a valuable source of diagnostic cancer biomarkers. Previous mass spectrometry (MS) studies have suggested that groups of related peptides discriminatory for different cancer types are generated ex vivo from abundant serum proteins by tumor-specific exopeptidases. We tested 2 complementary serum profiling strategies to see if similar peptides could be found that discriminate ovarian cancer from benign cases and healthy controls.
Methods: We subjected identically collected and processed serum samples from healthy volunteers and patients to automated polypeptide extraction on octadecylsilane-coated magnetic beads and separately on ZipTips before MALDI-TOF MS profiling at 2 centers. The 2 platforms were compared and case control profiling data analyzed to find altered MS peak intensities. We tested models built from training datasets for both methods for their ability to classify a blinded test set.
Results: Both profiling platforms had CVs of approximately 15% and could be applied for high-throughput analysis of clinical samples. The 2 methods generated overlapping peptide profiles, with some differences in peak intensity in different mass regions. In cross-validation, models from training data gave diagnostic accuracies up to 87% for discriminating malignant ovarian cancer from healthy controls and up to 81% for discriminating malignant from benign samples. Diagnostic accuracies up to 71% (malignant vs healthy) and up to 65% (malignant vs benign) were obtained when the models were validated on the blinded test set.
Conclusions: For ovarian cancer, altered MALDI-TOF MS peptide profiles alone cannot be used for accurate diagnoses.
An Oedometer for Studying Combined Effects of Temperature and Suction on Soils
Authors: François Bertrand , Laloui Lyesse 
Published: January 2010

Abstract
This paper reports the development of an advanced thermo-hydro-mechanical (THM) oedometer in order to characterize the behavior of soils under combined non-isothermal and unsaturated conditions. The simultaneous control of temperature, suction, and stress states within the sample required rigorous calibration. This THM oedometer accommodates samples 80 mm in diameter and 23 mm in height. The applied vertical stresses can reach values of up to 1 MPa, the controlled temperature ranges from 20 to 80°C, and the applied suction is up to 500 kPa. The paper thoroughly discusses the calibration of the device and presents some results of tests performed on sandy silt.
Use of Fe–31Mn–7.5Al–1.3Si–0.9C Alloy for Fabrication of Resistive Elements
Authors: Casteletti L. C. , Lombardi Neto A. , Totten G. E. , Heck S. C. , Fernandes F. A. P. 
Published: January 2010

Abstract
Austenitic Fe–Mn–Al–Si–C steel provides excellent cold formability and good oxidation resistance for possible fabrication of resistive elements. In this work, austenitic Fe–31Mn–7.5Al–1.3Si–0.9C steel wire was obtained by cold rolling and then tested with respect to oxidation resistance and electrical resistive behavior. After being subjected to an isothermal oxidation test at 700°C in air, this steel exhibited behavior similar to that the AISI 304 stainless steel. When the test was conducted at 800°C, the performance was inferior to that the AISI 304 steel. However, after 40 h of testing, the mass-gain curve stabilized. From these results and the results of the electric resistance tests, the Fe–Mn–Al–Si alloy proved to be a promising candidate for substitution of thermo-resistive alloys of Ni–Cr and Fe–Ni–Cr for use at temperatures of up to 800°C. Such alloys provide the potential for cost performance savings by using lower cost alloying elements.
Determination of ε-Caprolactam Concentration in Water and a White Wine Simulant during Simulated Transport and Storage of Lidded Cups
Authors: Ianneo J. C. , Brown J. W. , Vorst K. , Singh J. 
Published: January 2010

Abstract
Under normal sealing and storage conditions, nylon-6, poly (caprolactam)-based plastic laminates may release impurities to packaged foods and liquids, and the application of heat for cooking often increases the rate of migration. Epsilon-caprolactam is one of the main contaminates found to migrate from a nylon-6 poly (caprolactam) plastic film. The objectives of this study were to determine the effects of solvent, transportation, and storage time on the concentration of ε-caprolactam from a nylon-6-based lidding material in water or a white wine substitute (12 % ethanol) in poly (propylene) cups lidded with a nylon-6-based film. Cups were filled, sealed, packaged, stacked, and exposed to a simulated 3-day cross-country shipment. Cups were sampled with or without simulated shipment after 0, 7, 14, or 28 days at 20.6°C. Epsilon-caprolactam was determined using a gas chromatograph (GC) equipped with flame ionization detector and Restek Rtx 1301 megabore column. Results of the study indicated release of ε-caprolactam into containers at the time of sealing with significantly higher levels (4.42 ppm average) occurring in 12 % ethanol versus water (0.01 ppm average). After the cups were sealed, neither simulated cross-country shipment nor storage increased levels of ε-caprolactam in either solvent. It is not understood how the alcohol, whether as liquid or vapor, interacted with the lidding material to increase ε-caprolactam release at the time of sealing. No delamination of the poly (propylene) layer of the lidding material was observed when cups were sealed. Future research needs to be conducted to study the effects of alcohol, alcohol vapor, sealing time, and temperature on potential ε-caprolactam release.
Comprehensive Facial Rehabilitation Improves Function in People With Facial Paralysis: A 5-Year Experience at the Massachusetts Eye and Ear Infirmary
Authors: Lindsay Robin W. , Robinson Mara , Hadlock Tessa A. 
Published: January 2010

Background
The Facial Grading Scale (FGS) is a quantitative instrument used to evaluate facial function after facial nerve injury. However, quantitative improvements in function after facial rehabilitation in people with chronic facial paralysis have not been shown.

Objective
The objectives of this study were to use the FGS in a large series of consecutive subjects with facial paralysis to quantitatively evaluate improvements in facial function after facial nerve rehabilitation and to describe the management of chronic facial paralysis.

Design
The study was a retrospective review.

Methods
A total of 303 individuals with facial paralysis were evaluated by 1 physical therapist at a tertiary care facial nerve center during a 5-year period. Facial rehabilitation included education, neuromuscular training, massage, meditation-relaxation, and an individualized home program. After 2 months of home exercises, the participants were re-evaluated, and the home program was tailored as necessary. All participants were evaluated with the FGS before the initiation of facial rehabilitation, and 160 participants were re-evaluated after receiving treatment. All participants underwent the initial evaluation at least 4 months after the onset of facial paralysis; for 49 participants, the evaluation took place more than 3 years after onset.

Results
Statistically significant increases in FGS scores were seen after treatment (P&lt;.001, t test). The average initial score was 56 (SD=21, range=13–98), and the average score after treatment was 70 (SD=18, range=25–100).

Limitations
A limitation of this study was that evaluations were performed by only 1 therapist.

Conclusions
For 160 patients with facial paralysis, statistically significant improvements after facial rehabilitation were shown; the improvements appeared to be long lasting with continued treatment. The improvements in the FGS scores indicated that patients can successfully manage symptoms with rehabilitation and underscored the importance of specialized therapy in the management of facial paralysis.