bluuebunny/crossref_metadata_2025_split huggingface dataset:
Industrial-scale production and purification of a heterologous protein in Lactococcus lactis using the nisin-controlled gene expression system NICE: The case of lysostaphin
Authors: Mierau Igor , Leij Peter , van Swam Iris , Blommestein Barry , Floris Esther , Mond James , Smid Eddy J 
Published: May 2005

Abstract

Background
The NI sin-C ontrolled gene E xpression system NICE of Lactococcus lactis is one of the most widespread used expression systems of Gram-positive bacteria. It is used in more than 100 laboratories for laboratory-scale gene expression experiments. However, L. lactis is also a micro-organism with a large biotechnological potential. Therefore, the aim of this study was to test whether protein production in L. lactis using the NICE system can also effectively be performed at the industrial-scale of fermentation.

Results
Lysostaphin, an antibacterial protein (mainly against Staphylococcus aureus) from S. simulans biovar. Staphylolyticus, was used as a model system. Food-grade lysostaphin expression constructs in L. lactis were grown at 1L-, 300-L and 3000-L scale and induced with nisin for lysostaphin production. The induction process was equally effective at all scales and yields of about 100 mg/L were obtained. Up-scaling was easy and required no specific effort. Furthermore, we describe a simple and effective way of downstream processing to obtain a highly purified lysostaphin, which has been used for clinical phase I trials.

Conclusion
This is the first example that shows that nisin-regulated gene expression in L. lactis can be used at industrial scale to produce large amounts of a target protein, such as lysostaphin. Downstream processing was simple and in a few steps produced a highly purified and active enzyme.
Stepping stability: effects of sensory perturbation
Authors: McGibbon Chris A , Krebs David E , Wagenaar Robert 
Published: May 2005

AbstractBackgroundFew tools exist for quantifying locomotor stability in balance impaired populations. The objective of this study was to develop and evaluate a technique for quantifying stability of stepping in healthy people and people with peripheral (vestibular hypofunction, VH) and central (cerebellar pathology, CB) balance dysfunction by means a sensory (auditory) perturbation test.MethodsBalance impaired and healthy subjects performed a repeated bench stepping task. The perturbation was applied by suddenly changing the cadence of the metronome (100 beat/min to 80 beat/min) at a predetermined time (but unpredictable by the subject) during the trial. Perturbation response was quantified by computing the Euclidian distance, expressed as a fractional error, between the anterior-posterior center of gravity attractor trajectory before and after the perturbation was applied. The error immediately after the perturbation (Emax), error after recovery (Emin) and the recovery response (Edif) were documented for each participant, and groups were compared with ANOVA.ResultsBoth balance impaired groups exhibited significantly higher Emax (p= .019) and Emin (p= .028) fractional errors compared to the healthy (HE) subjects, but there were no significant differences between CB and VH groups. Although response recovery was slower for CB and VH groups compared to the HE group, the difference was not significant (p= .051).ConclusionThe findings suggest that individuals with balance impairment have reduced ability to stabilize locomotor patterns following perturbation, revealing the fragility of their impairment adaptations and compensations. These data suggest that auditory perturbations applied during a challenging stepping task may be useful for measuring rehabilitation outcomes.
Nonmelanoma Skin Cancer in Survivors of Childhood and Adolescent Cancer: A Report From the Childhood Cancer Survivor Study
Authors: Perkins Joanna L. , Liu Yan , Mitby Pauline A. , Neglia Joseph P. , Hammond Sue , Stovall Marilyn , Meadows Anna T. , Hutchinson Ray , Dreyer ZoAnn E. , Robison Leslie L. , Mertens Ann C. 
Published: May 2005

Purpose Nonmelanoma skin cancer (NMSC) has become the most common type of cancer in many populations throughout the world. Ultraviolet and ionizing radiation are known risk factors. Because NMSCs are rarely lethal and most cancer registries do not routinely report data regarding these cancers, they have received little attention in studies evaluating long-term effects of cancer therapy. This article reports on the occurrence of secondary NMSC as a long-term effect of cancer therapy in survivors of childhood cancer. Patients and Methods The Childhood Cancer Survivor Study (CCSS) is a cohort study of 5-year survivors of childhood and adolescent cancer from 25 participating institutions in North America. NMSC patients were defined by a history of basal cell or squamous cell carcinoma of the skin after primary malignancy treatment. Demographic and treatment data were collected and analyzed. Results Among the 13,132 eligible CCSS participants, 213 have reported NMSC; 99 patients (46%) have had multiple occurrences. Median age of occurrence was 31 years (range, 7 to 46 years). Location of NMSC included head and neck (43%), back (24%), chest (22%), abdomen and pelvis (5%), extremity (3%), and unknown (4%). Ninety percent of patients had previously received radiation therapy (RT); 90% of tumors occurred within the RT field. RT was associated with a 6.3-fold increase in risk (95% CI, 3.5- to 11.3-fold). Conclusion Long-term survivors of childhood and adolescent cancer who were treated with RT are at highest risk for developing NMSC. Educational efforts need to be directed to this population to facilitate early diagnosis of NMSC and reduction in sun exposure.
Analysis of Treatment Practices for Elderly Cancer Patients in Ontario, Canada
Authors: Townsley Carol , Pond Gregory R. , Peloza Brenda , Kok Joanne , Naidoo Kendra , Dale Darlene , Herbert Carole , Holowaty Eric , Straus Sharon , Siu Lillian L. 
Published: May 2005

Purpose Older patients are underrepresented in many areas of cancer services utilization and in clinical trial enrollment. This study evaluates whether age, when adjusted for sex, comorbidity, stage, tumor site, geography, and time period, is predictive of cancer treatment practice. Methods First, we used the Ontario Cancer Registry (OCR) to examine for any apparent differences in treatment practices between elderly (≥ 70 years) and younger patients in the last three decades. Second, we performed a chart review of 1,505 patients with lung, breast, and colorectal cancers seen in Ontario either at an urban center, the Princess Margaret Hospital, or at a rural center, the Northwestern Regional Cancer Centre. Patients were randomly selected from two time periods, 1977 to 1978 and 1997; and the study population was to comprise at least 50% elderly patients. Results OCR data demonstrated that, in some settings, such as colorectal cancer, the proportions of elderly cancer patients who were referred to cancer centers and who received any cancer treatment were lower than their younger counterparts. The chart review data showed that increasing age was a significant negative predictor for receiving any cancer treatment (P < .001, multivariate analysis) and for having a clinical trial discussion with the treating specialist (P < .001, multivariate analysis). Conclusion Independent of other factors, older age is consistently a cause of disparity in cancer treatment practice and in clinical trial discussion with patients. By increasing the accrual rate of elderly cancer patients in clinical trials, a better understanding of appropriate therapies for this patient population can be obtained and may, thereby, impact on their cancer-related morbidity and mortality.
Endogenous Ghrelin Regulates Episodic Growth Hormone (GH) Secretion by Amplifying GH Pulse Amplitude: Evidence from Antagonism of the GH Secretagogue-R1a Receptor
Authors: Zizzari P. , Halem H. , Taylor J. , Dong J. Z. , Datta R. , Culler M. D. , Epelbaum J. , Bluet-Pajot M. T. 
Published: May 2005

Abstract
Ghrelin was purified from rat stomach as an endogenous ligand for the GH secretagogue (GHS) receptor. As a GHS, ghrelin stimulates GH release, but it also has additional activities, including stimulation of appetite and weight gain. Plasma GH and ghrelin secretory patterns appear unrelated, whereas many studies have correlated ghrelin variations with food intake episodes. To evaluate the role of endogenous ghrelin, GH secretion and food intake were monitored in male rats infused sc (6 μg/h during 10 h) or intracerebroventricularly (5 μg/h during 48 h) with BIM-28163, a full competitive antagonist of the GHS-R1a receptor. Subcutaneous BIM-28163 infusion significantly decreased GH area under the curve during a 6-h sampling period by 54% and peak amplitude by 46%. Twelve hours after the end of treatment these parameters returned to normal. Central treatment was similarly effective (−37 and −42% for area under the curve and −44 and −49% for peak amplitude on the first and second days of infusion, respectively). Neither peripheral nor central BIM-28163 injection modified GH peak number, GH nadir, or IGF-I levels. In this protocol, food intake is not strongly modified and water intake is unchanged. Subcutaneous infusion of BIM-28163 did not change plasma leptin and insulin levels evaluated at 1200 and 1600 h. On the contrary, central BIM-28163 infusion slightly increased leptin and significantly increased insulin concentrations. Thus, endogenous ghrelin, through GHS-R1a, acts as a strong endogenous amplifier of spontaneous GH peak amplitude. The mechanisms by which ghrelin modifies food intake remain to be defined and may involve a novel GHS receptor.
Src Homology 3-Domain Growth Factor Receptor-Bound 2-Like (Endophilin) Interacting Protein 1, a Novel Neuronal Protein that Regulates Energy Balance
Authors: Trevaskis James , Walder Ken , Foletta Victoria , Kerr-Bayles Lyndal , McMillan Janine , Cooper Adrian , Lee Scott , Bolton Kristy , Prior Matthew , Fahey Richard , Whitecross Kate , Morton Gregory J. , Schwartz Michael W. , Collier Greg R. 
Published: May 2005

AbstractTo identify genes involved in the central regulation of energy balance, we compared hypothalamic mRNA from lean and obese Psammomys obesus, a polygenic model of obesity, using differential display PCR. One mRNA transcript was observed to be elevated in obese, and obese diabetic, P. obesus compared with lean animals and was subsequently found to be increased 4-fold in the hypothalamus of lethal yellow agouti (Ay/a) mice, a murine model of obesity and diabetes. Intracerebroventricular infusion of antisense oligonucleotide targeted to this transcript selectively suppressed its hypothalamic mRNA levels and resulted in loss of body weight in both P. obesus and Sprague Dawley rats. Reductions in body weight were mediated by profoundly reduced food intake without a concomitant reduction in metabolic rate. Yeast two-hybrid screening, and confirmation in mammalian cells by bioluminescence resonance energy transfer analysis, demonstrated that the protein it encodes interacts with endophilins, mediators of synaptic vesicle recycling and receptor endocytosis in the brain. We therefore named this transcript Src homology 3-domain growth factor receptor-bound 2-like (endophilin) interacting protein 1 (SGIP1). SGIP1 encodes a large proline-rich protein that is expressed predominantly in the brain and is highly conserved between species. Together these data suggest that SGIP1 is an important and novel member of the group of neuronal molecules required for the regulation of energy homeostasis.
Perinatal Exposure to Bisphenol-A Alters Peripubertal Mammary Gland Development in Mice
Authors: Muñoz-de-Toro Monica , Markey Caroline M. , Wadia Perinaaz R. , Luque Enrique H. , Rubin Beverly S. , Sonnenschein Carlos , Soto Ana M. 
Published: May 2005

AbstractDevelopmental exposure to estrogenic chemicals induces morphological, functional, and behavioral anomalies associated with reproduction. Humans are exposed to bisphenol-A (BPA), an estrogenic compound that leaches from dental materials and plastic food and beverage containers. The aim of the present study was to determine the effects of perinatal exposure to low, environmentally relevant doses of BPA [25 and 250 ng BPA/kg body weight (bw)·d] on the peripubertal development of the mammary gland. BPA exposure enhanced the mammary glands’ sensitivity to estradiol in ovariectomized CD-1 mice. In their intact 30-d-old littermates, the area and numbers of terminal end buds relative to the gland ductal area increased whereas their apoptotic activity decreased. There was a positive correlation between ductal length and the age at first proestrus; that was reduced as the BPA dose increased, suggesting that BPA exposure slows down ductal invasion of the stroma. There was also a significant increase of progesterone receptor-positive ductal epithelial cells that were localized in clusters, suggesting future branching points. Indeed, lateral branching was significantly enhanced at 4 months of age in mice exposed to 25 ng BPA /kg bw·d. In conclusion, perinatal exposure to environmentally relevant BPA doses results in persistent alterations in mammary gland morphogenesis. Of special concern is the increased terminal end bud density at puberty as well as the increased number of terminal ends reported previously in adult animals, as these two structures are the sites at which cancer arises in humans and rodents.
Calmodulin-Dependent Kinase IV Stimulates Vitamin D Receptor-Mediated Transcription
Authors: Ellison Tara I. , Dowd Diane R. , MacDonald Paul N. 
Published: May 2005

Abstract1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] promotes intestinal absorption of calcium primarily by binding to the vitamin D receptor (VDR) and regulating gene expression. 1,25-(OH)2D3 also exerts rapid actions at the cell membrane that include increasing intracellular calcium levels and activating protein kinase cascades. To explore potential cross talk between calcium signaling elicited by the nongenomic actions of 1,25-(OH)2D3 and the genomic pathway mediated by VDR, we examined the effects of activated Ca2+/calmodulin-dependent kinases (CaMKs) on 1,25-(OH)2D3/VDR-mediated transcription. Expression of a constitutively active form of CaMKIV dramatically stimulated 1,25-(OH)2D3-activated reporter gene expression in COS-7, HeLa, and ROS17/2.8 cell lines. Metabolic labeling studies indicated that CaMKIV increased VDR phosphorylation levels. In addition, CaMKIV increased the independent transcription activity of the VDR coactivator SRC (steroid receptor coactivator) 1, and promoted ligand-dependent interaction between VDR and SRC coactivator proteins in mammalian two-hybrid studies. The functional consequences of this multifaceted mechanism of CaMKIV action were revealed by reporter gene studies, which showed that CaMKIV and select SRC coactivators synergistically enhanced VDR-mediated transcription. These studies support a model in which CaMKIV signaling stimulates VDR-mediated transcription by increasing phosphorylation levels of VDR and enhancing autonomous SRC activity, resulting in higher 1,25-(OH)2D3-dependent interaction between VDR and SRC coactivators.
Modulation of the Expression and Transactivation of Androgen Receptor by the Basic Helix-Loop-Helix Transcription Factor Pod-1 through Recruitment of Histone Deacetylase 1
Authors: Hong Cheol Yi , Gong Eun-Yeung , Kim Kabsun , Suh Ji Ho , Ko Hyun-Mi , Lee Hyun Joo , Choi Hueng-Sik , Lee Keesook 
Published: May 2005

AbstractAndrogen receptor (AR) is important in male sexual differentiation and testicular function. Here, we demonstrate the regulation of AR expression and its transactivation by the basic helix-loop-helix (bHLH) transcription factor Pod-1, the expression of which in postnatal testis reciprocally coincides with the expression of AR. Pod-1 represses the promoter activity of AR, possibly through its E-box. An AR promoter region of 169 bp, which harbors one canonical E-box, is sufficient for the Pod-1-repression and bound by purified Pod-1 proteins. Pod-1 also suppresses the transactivation of AR. Transient transfection analyses of mammalian cells show that Pod-1 represses AR transactivation in a dose-dependent manner. Furthermore, yeast two-hybrid, glutathione-S-transferase-pull-down, and coimmunoprecipitation analyses reveal that Pod-1 directly associates with AR through its N-terminal region and through the DNA binding-hinge domain of AR. Interestingly, Pod-1 recruits histone deacetylase (HDAC)-1 to inhibit both promoter activity and transactivation of AR. Overexpression of HDAC1 further inhibits the Pod-1-mediated repressions and Pod-1 directly interacts with HDAC1. Furthermore, chromatin immunoprecipitation assay reveals that HDAC1 is recruited with Pod-1 to the endogenous AR promoter and the androgen-regulated Pem promoter. Taken together, these results suggest that Pod-1, which controls AR transcription and function, may play an important role in the development and function of the testis.
The REALAB Project: A New Method for the Formulation of Reference Intervals Based on Current Data
Authors: Grossi Enzo , Colombo Roberto , Cavuto Silvio , Franzini Carlo 
Published: May 2005

AbstractBackground: In a primary healthcare center concerned more with maintaining wellness than with diagnosing and monitoring illness, it is particularly important to compare patients’ results with reference intervals derived from a matched population by use of defined statistical methods.Methods: Laboratory results over a 3-year period (∼15 000 000 records; 197 350 individuals) were retrieved from our laboratory information system. An inclusion/exclusion procedure for individual patients was applied based on (a) presence of at least 1 of 23 previously defined “basic tests”; (b) only 1 measurement per test by the laboratory over the 3-year period; (c) for each test, absence of any abnormality in the correlated tests. Before the third step, correlations among quantities were assessed by a Spearman correlation matrix, comparing each of the 23 basic tests with all remaining tests by use of a novel multivariate algorithm.Results: The initial sample group (n = 197 350) was reduced stepwise by the selection criteria outlined above to 166 027, then to 93 649, and finally to 61 246 individuals constituting our reference sample group. Results from the last 2 groups were used to calculate sex-specific, and in some cases age-related, reference limits for the 23 basic tests and for 13 additional quantities. Reference limits were calculated throughout this study by nonparametric estimation of percentiles.Conclusion: Reference values derived by retrospective analysis of large samples of data obtained at a given institution are particularly suitable for the evaluation of results for the presenting patient population at that institution.
Szenarienspezifikation zur Systemverhaltensanalyse mit Petrinetzen (Scenario Specification for the Analysis of System Behaviour by Petri-Nets)
Authors: Einer Stefan 
Published: May 2005

Abstract
Die formale Technik „STOP — Spezifikationstechnik operationaler Prozesse“ dient dazu, Szenarien eines Systems und deren Abweichungspotenzial zu einem analysierbaren Verhaltensmodell des Systems zu synthetisieren. Dazu wurde eine bestimmte Form der Petrinetzmodellierung entwickelt, die hier vorgestellt sowie anhand ihrer praktischen Anwendung veranschaulicht wird.
P- und PD-Polvorgaberegler für Regelstrecken mit begrenzter Stellgröße (P and PD Pole Assignment Controllers for Constrained Systems)
Authors: Huba Mikuláš 
Published: May 2005

Abstract
In diesem Beitrag werden zwei neue Verfahren zum Entwurf von Polvorgabereglern mit beschränkter Stellgröße für Regelstrecken mit einer dominanten Dynamik zweiter Ordnung betrachtet. Die Verfahren liefern einfache und zugleich zuverlässige Entwurfsergebnisse. Sie sind vor allem für die Regelung von instabilen Regelstrecken und für hohe Qualitätsansprüche an das dynamische Verhalten und an den stationären Zustand des Regelkreises bestimmt. Zuerst wird gezeigt, dass bei der Regelung von Strecken erster Ordnung die Stellgrößenbeschränkung zu keinen parasitären Nebenwirkungen wie Überschwingen, Oszillationen oder Instabilität führt. Im Fall von Strecken höherer Ordnung versucht man daher, die Regelungsaufgabe auf mehrere Teilaufgaben erster Ordnung zurückzuführen. Die Lösung dieser Aufgabe ist im Fall von reellen Polen möglich (obwohl nicht eindeutig).
Im Fall von komplexen Polen wird eine prinzipiell andere Möglichkeit genutzt. Durch eine nichtlineare Anpassung des Sollwertes wird die Zustandstrajektorie an eine optimale Bremskurve angenähert. Dies kann durch einfaches Gain-Scheduling ergänzt werden, um mögliche Zeitverzögerungen des Regelkreises in der Beschleunigungsphase zu kompensieren. Kombiniert man solche Reglerentwürfe, die für den doppelten Integrator durchgeführt wurden, mit einer Approximation der Regelstrecken durch I2PT1 oder I2Tt-Glieder, dann erhält man eine einfache und leicht benutzbare Methode für den Reglerentwurf mit Berücksichtigung von Begrenzungen und Zeitverzögerungen, die als eine Erweiterung des Verfahrens nach Ziegler und Nichols angesehen werden kann.
Ermittlung des Nutzens von Datenbankreorganisationen: Notwendigkeit, Werkzeuge, Herangehensweisen (Ascertaining the Benefit of Database Reorganizations: Need, Tools, Procedures)
Authors: Dorendorf Stefan 
Published: May 2005

Zusammenfassung
Um auch bei einer großen Menge zu verwaltender Daten einen hohen Durchsatz und ein gutes Antwortzeitverhalten zu bieten, werden bei modernen Datenbank-Management-Systemen bei Update-Operationen die physischen Strukturen zur Speicherung von Daten und Zugriffspfaden oftmals von den Produkten nicht sofort oder nicht vollständig gepflegt. Damit entstehen sukzessive Degenerierungen in den Strukturen, die sich negativ auf Speicherauslastung und Systemleistung auswirken können. Solche Degenerierungen können mit Datenbankreorganisationen beseitigt werden. Bevor jedoch eine solche meist aufwändige und die laufende Datenbankverarbeitung möglicherweise behindernde Reorganisation durchgeführt wird, ist es notwendig, Degenerierungen zu erkennen und zu lokalisieren. Weiterhin ist es wünschenswert, die Auswirkungen der Degenerierungen und den durch eine Reorganisation erreichbaren Nutzen einschätzen (quantifizieren) zu können. Dieser Beitrag beleuchtet einige Aspekte des Themengebietes Datenbankreorganisation und zeigt Vorgehensweisen, die im Rahmen von Reorganisationsbedarfsanalysen und Reorganisationsnutzenanalysen angewendet werden können, und betrachtet dabei auch derzeit am Markt verfügbare Produkte.
External Egg Membrane Characteristics of Landlocked Fall Chinook Salmon Eyed Eggs Treated with Hydrogen Peroxide or Formalin
Authors: Stephenson Hans , Gabel Mark , Barnes Michael E 
Published: May 2005

Abstract
This study examines the microscopic characteristics of external egg membranes from eyed eggs of landlocked fall Chinook salmon Oncorhynchus tshawytscha treated with formalin (1,667 mg/L) or hydrogen peroxide (700 mg/L) for 15 min daily until hatching. Just before the start of hatching, the internus layer became significantly less differentiated in the membranes of eggs treated with hydrogen peroxide compared with those in the formalin treatments or untreated controls. There were no significant differences among the treatments in total membrane thickness, pore canal height, basal layer height, pore canal width, and the number of pore canals per 50-μm lengthwise segment along the membrane. Overall membrane thickness (externus and internus layers combined) averaged 51.5 μm during the test period. Thickness of externus averaged 0.69 μm, nearly two times that reported previously for nonlandlocked Chinook salmon.