bluuebunny/crossref_metadata_2025 huggingface dataset:
IgG Subclass Concentrations in Certified Reference Material 470 and Reference Values for Children and Adults Determined with The Binding Site Reagents
Authors: Schauer Uwe , Stemberg Frank , Rieger Christian H L , Borte Michael , Schubert Simone , Riedel Frank , Herz Udo , Renz Harald , Wick Manfred , Carr-Smith Hugh D , Bradwell Arthur R , Herzog Wilhelm
Published: February 2004

Background: There is currently no international reference preparation for IgG subclass (IgGSc) quantification. This situation has led to calibration differences among assays and a variety of reference interval values with consequential difficulties in comparing results. We therefore evaluated IgGSc concentrations in Certified Reference Material 470 (CRM 470).

Methods: Pure, polyclonal IgG1, -2, -3, and -4 were prepared from a large serum pool for use as primary standards. The IgG mass in each preparation was calculated from amino-acid analysis data. IgGSc concentrations were assessed in CRM 470 by nephelometry with modern analytical techniques, using these reference preparations. Subsequently, IgGSc concentrations were measured in 380 healthy individuals (250 males and 130 females), and age-dependent reference intervals were established.

Results: IgGSc concentrations in CRM 470 were as follows: IgG1, 5028 mg/L; IgG2, 3418 mg/L; IgG3, 579 mg/L, and IgG4, 381 mg/L, with a total IgG concentration of 9406 mg/L, 2.83% below the certified total IgG value of 9680 mg/L. Age-dependent percentile curves for the four IgGSc were constructed using a Box–Cox transformation. Maximum median values were as follows: IgG1, 6.02 g/L at 11 years; IgG2, 3.45 g/L at 31 years; IgG3, 0.63 g/L at 17 years; and IgG4, 0.48 g/L at 14 years. No significant sex-related differences were observed.

Conclusions: The correlation between the summation of individual IgGSc and separate measurements of total IgG concentrations was good and supports the accuracy of the results. The results are based on The Binding Site assays and should not be considered appropriate for other assays unless so demonstrated.
Improved HPLC Method for Carbohydrate-deficient Transferrin in Serum
Authors: Helander Anders , Husa Asgeir , Jeppsson Jan-Olof
Published: February 2004

Background: There is need for a reference method for transferrin glycoforms in serum to which routine immunologic methods for the alcohol marker carbohydrate-deficient transferrin (CDT) can be traceable. We describe an improved HPLC method for transferrin glycoforms.

Methods: Transferrin was iron-saturated by mixing the serum with ferric nitrilotriacetic acid, and lipoproteins were precipitated with dextran sulfate and calcium chloride. Separation of glycoforms was performed on a SOURCE 15Q anion-exchange column using salt gradient elution. Quantification relied on selective absorbance of the iron–transferrin complex at 470 nm. The relative amount of each glycoform was calculated as a percentage of the area under the curve, using baseline integration.

Results: The HPLC system provided reproducible separation and quantification of the asialo-, monosialo-, disialo-, trisialo-, tetrasialo-, pentasialo-, and hexasialotransferrin glycoforms. Most importantly, disialo- and trisialotransferrin were almost baseline separated. The intra- and interassay CV for disialotransferrin were <5%. Serum and the pretreated samples were stable for at least 2 days at 22 or 4 °C. Sera from 132 healthy controls contained [mean (SD)] 1.16 (0.25)% disialotransferrin, 4.77 (1.36)% trisialotransferrin, 80.18 (2.01)% tetrasialotransferrin, and 13.88 (1.69)% pentasialo- + hexasialotransferrin. In some cases of a high (>6%) trisialotransferrin, monosialotransferrin was detected at <0.25%. Asialotransferrin was not detected in control sera, but was detected in 57% of chronic heavy drinkers and in 62% of sera with ≥2% disialotransferrin.

Conclusions: The HPLC method fulfills the requirements of a preliminary reference method for CDT and should work for any combination of serum transferrin glycoforms. This method could also be useful for confirming positive CDT results by immunoassays in medico-legal cases.
Evaluation of Oligonucleotide Arrays for Sequencing of the p53 Gene in DNA from Formalin-Fixed, Paraffin-Embedded Breast Cancer Specimens
Authors: Cooper Melissa , Li Shu-Qiu , Bhardwaj Tajinder , Rohan Thomas , Kandel Rita A
Published: February 2004

Background: Routine tissue processing has generated banks of paraffin-embedded tissue that could be used in retrospective cohort studies to study the molecular changes that occur during cancer development. The purpose of this study was to determine whether a p53 microarray could be used to sequence the p53 gene in DNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissues.

Methods: DNA was extracted from 70 FFPE breast cancer tissue specimens. p53 was sequenced with an oligonucleotide microarray (p53 GeneChip®; Affymetrix), and the results were compared with the results obtained from direct sequencing.

Results: DNA was extracted from 62 of 70 cases. We identified 26 mutations in 24 of the 62 cases by the p53 GeneChip. No polymorphisms were detected, and exon 4 could not be evaluated in 20 cases. There were 43 genetic alterations detected by direct sequencing in 35 of the 62 cases. These consisted of 26 polymorphisms and 17 mutations in exons or splice sites. Fifteen mutations were identified by both methods. Direct sequencing detected significantly more gene alterations (43 of 54) in DNA extracted from FFPE tissue than the p53 GeneChip (26 of 54; P = 0.018). However, if the changes in exon 4 were eliminated from this comparison, the p53 GeneChip detected 26 of 27 mutations compared with direct sequencing, which identified 16 of 27 mutations. (P = 0.016).

Conclusions: A combination of oligonucleotide microarray and direct sequencing may be necessary to accurately identify p53 gene alterations in FFPE breast cancer. The p53 GeneChip cannot be used to detect exon 4 polymorphisms (codon 72) in FFPE breast cancer tissue.
High Preoperative CA 15-3 Concentrations Predict Adverse Outcome in Node-Negative and Node-Positive Breast Cancer: Study of 600 Patients with Histologically Confirmed Breast Cancer
Authors: Duffy Michael J , Duggan Catherine , Keane Rachel , Hill Arnold D K , McDermott Enda , Crown John , O’Higgins Niall
Published: February 2004

Background: CA 15-3 is the most widely used serum marker in breast cancer. Currently, its main uses are in the surveillance of patients with diagnosed disease and monitoring the treatment of patients with advanced disease.

Methods: Preoperative CA 15-3 concentrations were measured prospectively in 600 patients with histologically confirmed breast cancer. Marker concentrations were related to patient outcome by both univariate and multivariate analysis.

Results: After a median follow-up of 6.27 years, patients with high preoperative concentrations of CA 15-3 (>30 units/L) had a significantly shorter overall survival pattern than those with low concentrations. As a prognostic factor, CA 15-3 was independent of tumor size, axillary node status, and patient age. As well as being prognostic in the total population of patients, CA 15-3 also predicted outcome in different subgroups of patients, including those with both node-negative and node-positive disease, those who were both estrogen receptor (ER)-negative and ER-positive, and those younger and older that 50 years of age. CA 15-3 was also predictive of outcome irrespective of the type of adjuvant therapy administered, i.e., whether adjuvant hormone therapy, adjuvant chemotherapy, or radiotherapy was administered.

Conclusion: Assay of CA 15-3 is a relatively inexpensive, convenient, and noninvasive method for evaluating prognosis in newly diagnosed breast cancer patients.
In Vitro Contamination of Hair by Marijuana Smoke
Authors: Thorspecken Jana , Skopp Gisela , Pötsch Lucia
Published: February 2004

Background: The deposition of cannabinoids on/into hair from environmental smoke can be considered as a potential source of drug findings in hair. We studied external uptake of cannabinoids from marijuana smoke, investigating possible influencing factors on drug uptake and the efficiency of decontamination procedures.

Methods: Strands of a natural hair sample were moistened with water, greased with sebum or sebum/sweat, or bleached or permed. Treated and untreated samples were exposed to marijuana smoke for 60 min. Aliquots of each hair strand were either kept unwashed or were washed with methanol, dichloromethane, or 5 g/L dodecyl sulfate in water. Cannabinoid concentrations in unwashed and washed hair samples, as well as in air samples collected from the exposure chamber and in the marijuana sample being combusted, were quantified by gas chromatography–mass spectrometry or gas chromatography.

Results: Cannabinoids were deposited on the hair fibers from marijuana smoke. Cannabinoid concentrations were dependent on air concentration and hair pretreatment. Uptake was less in untreated than in pretreated hair. Concentrations were increased in damp hair, but were even higher in greased hair. There was no significant difference in concentration between bleached and permed strands. External contaminants were completely removed by washing with methanol and dichloromethane in untreated hair only. Washing with dodecyl sulfate in water was insufficient in all cases.

Conclusions: Exposures of hair to marijuana smoke yields detectable cannabinoids depending on concentrations in the air, hair care habits, and cosmetic treatment. Environmental marijuana smoke exposure may produce false-positive or falsely increased test results in hair.
Properties of the Reverse Transcription Reaction in mRNA Quantification
Authors: Ståhlberg Anders , Håkansson Joakim , Xian Xiaojie , Semb Henrik , Kubista Mikael
Published: February 2004

Background: In most measurements of gene expression, mRNA is first reverse-transcribed into cDNA. We studied the reverse transcription reaction and its consequences for quantitative measurements of gene expression.

Methods: We used SYBR green I-based quantitative real-time PCR (QPCR) to measure the properties of reverse transcription reaction for the β-tubulin, glyceraldehyde-3-phosphate dehydrogenase, Glut2, CaV1D, and insulin II genes, using random hexamers, oligo(dT), and gene-specific reverse transcription primers.

Results: Experimental variation in reverse transcription-QPCR (RT-QPCR) was mainly attributable to the reverse transcription step. Reverse transcription efficiency depended on priming strategy, and the dependence was different for the five genes studied. Reverse transcription yields also depended on total RNA concentration.

Conclusions: RT-QPCR gene expression measurements are comparable only when the same priming strategy and reaction conditions are used in all experiments and the samples contain the same total amount of RNA. Experimental accuracy is improved by running samples in (at least) duplicate starting with the reverse transcription reaction.
Differential Gene Expression of Eph Receptors and Ephrins in Benign Human Tissues and Cancers
Authors: Hafner Christian , Schmitz Gerd , Meyer Stefanie , Bataille Frauke , Hau Peter , Langmann Thomas , Dietmaier Wolfgang , Landthaler Michael , Vogt Thomas
Published: February 2004

Background: Eph receptors and their ligands, the ephrins, represent a large class of cell–cell communication molecules with well-defined developmental functions. Their role in healthy adult tissues and in human disease is still largely unknown, although diverse roles in carcinogenesis have been postulated.

Methods: We established a set of fluorescent PCR probes and primers for the definition of individual gene expression profiles of 12 different Eph receptors and 8 ephrins in 13 different healthy tissues. The mRNA expression profiles were studied in human lung, colorectal, kidney, liver, and brain cancers.

Results: The family of Eph receptors/ephrins was widely expressed in adult tissues with organ-site-specific patterns: EphB6 was highest in the thymus, compatible with an involvement in T-cell maturation. Brain and testis shared a unique pattern with EphA6, EphA8, and EphB1 being the most prominent. EphA7 had a high abundance in the kidney vasculature. Ephrin-A3 was up-regulated 26-fold in lung cancer, and EphB2 was up-regulated 9-fold in hepatocellular carcinoma. EphA8 was down-regulated in colon cancer, and EphA1/EphA8 was down-regulated in glioblastomas.

Conclusion: Eph/Ephrin genes are widely expressed in all adult organs with certain organ-site-specific patterns. Because their function in adult tissues remains unknown, further analysis of their role in disease may disclose new insights beyond their well-defined meaning in development.
Improved Specificity of Newborn Screening for Congenital Adrenal Hyperplasia by Second-Tier Steroid Profiling Using Tandem Mass Spectrometry
Authors: Lacey Jean M , Minutti Carla Z , Magera Mark J , Tauscher Angela L , Casetta Bruno , McCann Mark , Lymp James , Hahn Si Houn , Rinaldo Piero , Matern Dietrich
Published: February 2004

Background: Newborn screening for congenital adrenal hyperplasia (CAH) involves measurement of 17α-hydroxyprogesterone (17-OHP), usually by immunoassay. Because this testing has been characterized by high false-positive rates, we developed a steroid profiling method that uses liquid chromatography–tandem mass spectrometry (LC-MS/MS) to measure 17-OHP, androstenedione, and cortisol simultaneously in blood spots.

Methods: Whole blood was eluted from a 4.8-mm (3/16-inch) dried-blood spot by an aqueous solution containing the deuterium-labeled internal standard d8-17-OHP. 17-OHP, androstenedione, and cortisol were extracted into diethyl ether, which was subsequently evaporated and the residue dissolved in LC mobile phase. This extract was injected into a LC-MS/MS equipped with pneumatically assisted electrospray. The steroids were quantified in the selected-reaction monitoring mode by use of peak areas in reference to the stable-isotope-labeled internal standard. We analyzed 857 newborn blood spots, including 14 blood spots of confirmed CAH cases and 101 of false-positive cases by conventional screening.

Results: Intra- and interassay CVs for 17-OHP were 7.2–20% and 3.9–18%, respectively, at concentrations of 2, 30, and 50 μg/L. At a cutoff for 17-OHP of 12.5 μg/L and a cutoff of 3.75 for the sum of peak areas for 17-OHP and androstenedione divided by the peak area for cortisol, 86 of the 101 false-positive samples were within reference values by LC-MS/MS, whereas the 742 normal and 14 true-positive results obtained by conventional screening were correctly classified.

Conclusion: Steroid profiling in blood spots can identify false-positive results obtained by conventional newborn screening for CAH.
Prognostic Value of Tubular Proteinuria and Enzymuria in Nonoliguric Acute Tubular Necrosis
Authors: Herget-Rosenthal Stefan , Poppen Dennis , Hüsing Johannes , Marggraf Günter , Pietruck Frank , Jakob Heinz-Günther , Philipp Thomas , Kribben Andreas
Published: February 2004

Background: Acute tubular necrosis (ATN) has high mortality, especially in patients who require renal replacement therapy (RRT). We prospectively studied the diagnostic accuracy of the urinary excretion of low-molecular-weight proteins and enzymes as predictors of a need for RRT in ATN.

Methods: In 73 consecutive patients with initially nonoliguric ATN, we measured urinary excretion of α1- and β2-microglobulin, cystatin C, retinol-binding protein, α-glutathione S-transferase, γ-glutamyltransferase, lactate dehydrogenase, and N-acetyl-β-d-glucosaminidase early in the course of ATN.

Results: Twenty-six patients (36%) required RRT a median of 4 (interquartile range, 2–6) days after detection of proteinuria and enzymuria. Patients who required RRT had higher urinary cystatin C and α1-microglobulin [median (interquartile range), 1.7 (1.2–4.1) and 34.5 (26.6–45.1) g/mol of creatinine] than patients who did not require RRT [0.1 (0.02–0.5) and 8.0 (5.0–17.5) g/mol of creatinine]. Urinary excretion of cystatin C and α1-microglobulin had the highest diagnostic accuracies in identifying patients requiring RRT as indicated by the largest areas under the ROC curves: 0.92 (95% confidence interval, 0.86–0.96) and 0.86 (0.78–0.92), respectively. Sensitivity and specificity were 92% (95% confidence interval, 83–96%) and 83% (73–90%), respectively, for urinary cystatin C >1 g/mol of creatinine, and 88% (78–93%) and 81% (70–88%) for urinary α1-microglobulin >20 g/mol of creatinine.

Conclusion: In nonoliguric ATN, increased urinary excretion of cystatin C and α1-microglobulin may predict an unfavorable outcome, as reflected by the requirement for RRT.
Mass Spectrometric Phenotyping of Val34Leu Polymorphism of Blood Coagulation Factor XIII by Differential Peptide Display
Authors: Tammen Harald , Möhring Thomas , Kellmann Markus , Pich Andreas , Kreipe Hans H , Hess Rüdiger
Published: February 2004

Background: The Val34Leu mutation in the activation peptide of factor XIII (FXIIIA) correlates with a lower incidence of myocardial infarction and ischemic stroke but an increased risk for hemorrhagic stroke. We describe mass spectrometric detection of the activation peptide variants in human serum.

Methods: We used differential peptide display (DPD) to compare comprehensive peptide maps from pairs of serum samples from healthy volunteers. Peptides were separated by liquid chromatography, and fractions were subjected to mass spectrometry. Mass spectra of all fractions were combined, giving a peptide map representing a two-dimensional display of peptide masses. After comparison of peptide mass maps, peptides that differentiated FXIIIA phenotypes were identified by mass spectrometry.

Results: Val34Leu polymorphisms of the activation peptide of FXIIIA were identified in 20 serum samples from 10 volunteers by DPD, and their sequences were confirmed by nanoelectrospray-ionization quadrupole time-of-flight mass spectrometry. Analysis of three (V34V, V34L, and L34L) phenotypes was confirmed by allele-specific genotypic analysis in all (n = 10) volunteers.

Conclusion: DPD provides a simple and easy-to-use phenotype assay with advantages over PCR-based assays in being faster and directly analyzing the compound of interest.
Association of Coronary Heart Disease with Pre-β-HDL Concentrations in Japanese Men
Authors: Hattori Hiroaki , Kujiraoka Takeshi , Egashira Tohru , Saito Eiji , Fujioka Takayuki , Takahashi Sadao , Ito Mayumi , Cooper Jackie A , Stepanova Irina P , Nanjee M Nazeem , Miller Norman E
Published: February 2004

Background: In individuals heterozygous for ABCA1 transporter mutations, defective reverse cholesterol transport (RCT) causes low HDL-cholesterol and premature coronary heart disease (CHD). However, the extent to which impaired RCT underlies premature CHD in others with low HDL-cholesterol is not known. The primary acceptors of cell cholesterol are a minor subclass of lipid-poor pre-β-HDLs. These are generated during remodeling of α-HDLs, which account for almost all HDL-cholesterol. We studied the strength of the association of CHD with pre-β-HDL concentrations in Japanese men.

Methods: Blood was collected from 42 men with clinical CHD and 44 healthy controls 40–70 years of age. Pre-β-HDL was assayed by crossed immunoelectrophoresis.

Results: Cases had lower HDL-cholesterol (−23%), total apolipoprotein A-I (−26%), and pre-β-HDL (−55%; all P <0.001) concentrations; lower pre-β-HDL:α-HDL ratios (−45%; P <0.001); and higher plasma triglycerides (20%; P <0.03) than the controls. On stepwise logistic regression, CHD was associated most strongly with pre-β-HDL concentrations. On ROC analysis, pre-β-HDL concentration discriminated between cases and controls better than any other lipoprotein measurement. When plasma was incubated for 16 h at 37 °C, mean (SD) pre-β-HDL increased by 47 (36)% in controls, but was unchanged in cases (group difference, P <0.001).

Conclusions: Our results suggest that inefficient RCT, secondary to a low pre-β-HDL concentration and production rate in plasma, contributes to premature CHD in Japanese men with low HDL-cholesterol.
Functional Reorganization of Visual Cortex Maps after Ischemic Lesions Is Accompanied by Changes in Expression of Cytoskeletal Proteins and NMDA and GABAAReceptor Subunits
Authors: Zepeda Angelica , Sengpiel Frank , Guagnelli Miguel Angel , Vaca Luis , Arias Clorinda
Published: February 2004

Reorganization of cortical representations after focal visual cortex lesions has been documented. It has been suggested that functional reorganization may rely on cellular mechanisms involving modifications in the excitatory/inhibitory neurotransmission balance and on morphological changes of neurons peripheral to the lesion. We explored functional reorganization of cortical retinotopic maps after a focal ischemic lesion in primary visual cortex of kittens using optical imaging of intrinsic signals. After 1, 2, and 5 weeks postlesion (wPL), we addressed whether functional reorganization correlated in time with changes in the expression of MAP-2, GAP-43, GFAP, GABAAreceptor subunit α1 (GABAAα1), subunit 1 of the NMDA receptor (NMDAR1), and in neurotransmitter levels at the border of the lesion. Our results show that: (1) retinotopic maps reorganize with time after an ischemic lesion; (2) MAP-2 levels increase gradually from 1wPL to 5wPL; (3) MAP-2 upregulation is associated with an increase in dendritic-like structures surrounding the lesion and a decrease in GFAP-positive cells; (4) GAP-43 levels reach the highest point at 2wPL; (5) NMDAR1 and glutamate contents increase in parallel from 1wPL to 5wPL; (6) GABAAα1 levels increase from 1wPL to 2wPL but do not change after this time point; and (7) GABA contents remain low from 1wPL to 5wPL. This is a comprehensive study showing for the first time that functional reorganization correlates in time with dendritic sprouting and with changes in the excitatory/inhibitory neurotransmission systems previously proposed to participate in cortical remodeling and suggests mechanisms by which plasticity of cortical representations may occur.
Dynamic Influences on Coincidence Detection in Neocortical Pyramidal Neurons
Authors: Grande Lucinda A. , Kinney Gregory A. , Miracle Greta L. , Spain William J.
Published: February 2004

The firing rate of neocortical pyramidal neurons is believed to represent primarily the average arrival rate of synaptic inputs; however, it has also been found to vary somewhat depending on the degree of synchrony among synaptic inputs. We investigated the ability of pyramidal neurons to perform coincidence detection, that is, to represent input timing in their firing rate, and explored some factors that influence that representation. We injected computer-generated simulated synaptic inputs into pyramidal neurons during whole-cell recordings, systematically altering the phase delay between two groups of periodic simulated input events. We explored how input intensity, the synaptic time course, inhibitory synaptic conductance, and input jitter influenced the firing rate representation of input timing. In agreement with computer modeling studies, we found that input synchronization increases firing rate when intensity is low but reduces firing rate when intensity is high. At high intensity, the effect of synchrony on firing rate could be switched from reducing to increasing firing rate by shortening the simulated excitatory synaptic time course, adding inhibition (using the dynamic clamp technique), or introducing a small input jitter. These opposite effects of synchrony may serve different computational functions: as a means of increasing firing rate it may be useful for efficient recruitment or for computing a continuous parameter, whereas as a means of decreasing firing rate it may provide gain control, which would allow redundant or excessive input to be ignored. Modulation of dynamic input properties may allow neurons to perform different operations depending on the task at hand.
Impaired Recency Judgments and Intact Novelty Judgments after Fornix Transection in Monkeys
Authors: Charles David P. , Gaffan David , Buckley Mark J.
Published: February 2004

Over four experiments based on the delayed matching-to-sample task, fornix-transected and normal control monkeys were presented with a sequence of five sample stimuli and then received intermixed within-session recency (WSR) and between-session recency (BSR) tests in experiment 1, only BSR tests in experiment 2, only absolute novelty (AN) tests in experiment 3, or only WSR tests in experiment 4. In WSR tests, monkeys chose which of two samples had occurred more recently in the immediately preceding sequence. In BSR and AN tests, monkeys were required to choose one sample from the immediately preceding sequence in preference to a foil unseen in the present session (BSR) or an AN foil that had never been presented before. When tests of WSR and BSR were intermixed (experiment 1), fornix monkeys performed below the level of the control monkeys in both types of test, although this difference was not statistically significant. In experiment 2, fornix monkeys were significantly impaired on tests of BSR alone, in which memory for a stimulus presented in an immediately preceding sequence could compete with memory for a foil presented in an earlier training session. In tests of AN (experiment 3), fornix monkeys performed at the same level as control animals in distinguishing a previously experienced stimulus from a previously unseen foil. In experiment 4, fornix transection significantly impaired tests of WSR alone. Taken together, these results suggest that one specialized role of the fornix is to process temporal information.
Clearance of α-Synuclein Oligomeric Intermediates via the Lysosomal Degradation Pathway
Authors: Lee He-Jin , Khoshaghideh Farnaz , Patel Smita , Lee Seung-Jae
Published: February 2004

Cytoplasmic deposition of α-synuclein aggregates is a common pathological feature of many neurodegenerative diseases. Strong evidence for the causative role of α-synuclein in these disorders is provided by genetic linkage between this gene and familial Parkinson's disease and by neurodegeneration in transgenic animals that overexpress this protein. In particular, it has been hypothesized that the accumulation of nonfibrillar oligomers of α-synuclein, which serve as intermediates for fibrillar inclusion body formation, causes neurodegeneration. However, little is known about how cells handle potentially toxic protein aggregates. Here we demonstrate that cells are capable of clearing preformed α-synuclein aggregates via the lysosomal degradation pathway. Consequently, blocking this pathway causes the accumulation of the aggregates in non-neuronal cells, differentiated neuroblastoma cells, and primary cortical neurons. This aggregate clearance occurs in an aggregation stage-specific manner; oligomeric intermediates are susceptible to clearance, whereas mature fibrillar inclusion bodies are not. Neutralization of the acidic compartments leads to the accumulation of α-synuclein aggregates and exacerbates α-synuclein toxicity in postmitotic neuronal cells, suggesting that the accumulation of oligomeric intermediates may be an important event leading to α-synuclein-mediated cell death. These results suggest that enhancing lysosomal function may be a potential therapeutic strategy to halt or even prevent the pathogenesis of Parkinson's disease and other Lewy body diseases.
Oxidized Galectin-1 Stimulates Macrophages to Promote Axonal Regeneration in Peripheral Nerves after Axotomy
Authors: Horie Hidenori , Kadoya Toshihiko , Hikawa Naoshi , Sango Kazunori , Inoue Hiroko , Takeshita Kaori , Asawa Reiko , Hiroi Tomoko , Sato Manami , Yoshioka Tohru , Ishikawa Yoshihiro
Published: February 2004

Various neurotrophic factors that promote axonal regeneration have been investigatedin vivo, but the signals that prompt neurons to send out processes in peripheral nerves after axotomy are not well understood. Previously, we have shown oxidized galectin-1 (GAL-1/Ox) promotes initial axonal growth after axotomy in peripheral nerves. However, the mechanism by which GAL-1/Ox promotes axonal regeneration remains unclear and is the subject of the present study. To identify possible target cells of GAL-1/Ox, a fluorescently labeled recombinant human GAL-1/Ox (rhGAL-1/Ox) was incubated with DRG neurons, Schwann cells, and intraperitoneal macrophages from adult rats. Only the cell surfaces of intraperitoneal macrophages bound the rhGAL-1/Ox, suggesting that these cells possess a receptor for GAL-1/Ox. Experiments examining tyrosine phosphorylation revealed that rhGAL-1/Ox stimulated changes in signal transduction pathways in these macrophages. These changes caused macrophages to secrete an axonal growth-promoting factor. This was demonstrated when conditioned media of macrophages stimulated with rhGAL-1/Ox in 48 hr culture strongly enhanced axonal regeneration from transected-nerve sites of DRG explants. Furthermore, activated macrophage-conditioned media also improved Schwann cell migration from the transected-nerve sites. From these results, we propose that axonal regeneration occurs in axotomized peripheral nerves as a result of cytosolic reduced galectin-1 being released from Schwann cells and injured axons, which then becomes oxidized in the extracellular space. Oxidized galectin-1 then stimulates macrophages to secrete a factor that promotes axonal growth and Schwann cell migration, thus enhancing peripheral nerve regeneration.
Inducible cAMP Early Repressor Regulates Corticosterone Suppression after Tricyclic Antidepressant Treatment
Authors: Conti Alana C. , Kuo Yuo-Chen , Valentino Rita J. , Blendy Julie A.
Published: February 2004

The cAMP-response element binding protein (CREB) is involved in antidepressant action, but the role of related CRE-binding transcription factors in the behavioral and endocrine responses to antidepressants is unclear. Alternative transcription of the cAMP response element-modulator (CREM) gene yields activator and repressor isoforms, including the strong repressor inducible cAMP early repressor (ICER). ICER is highly expressed in hypothalamic tissues and upregulated after electroconvulsive seizure. Thus, ICER may be a novel mediator of antidepressant action at endocrine and/or behavioral levels. Here we establish that both subchronic and chronic desipramine (DMI) treatments upregulate hypothalamic ICER expression in wild-type mice. Behavioral responses to DMI in the forced swim and tail suspension tests are unchanged in mice lacking ICER. However, the ability of DMI to suppress an acute corticosterone response after swim stress is compromised in ICER-deficient mice, suggesting that increased hypothalamic ICER mRNA after DMI treatment may be required for suppression of corticosterone release. To investigate the mechanism underlying this response, we measured corticotropin releasing factor (CRF), an upstream modulator of corticosterone release. Using real-time quantitative PCR, we establish that hypothalamic CRF expression is significantly reduced after swim exposure in DMI-treated wild-type mice, however DMI is unable to blunt hypothalamic CRF expression in ICER-deficient mice. Furthermore, we demonstrate that ICER is enriched in CRF-expressing neurons in the paraventricular nucleus of the hypothalamus. These data indicate that ICER is required for DMI to reduce stress-induced corticosterone release through regulation of hypothalamic CRF expression, revealing a novel role for ICER in antidepressant regulation of the hypothalamic-pituitary adrenal axis.
Essential Function of Oncostatin M in Nociceptive Neurons of Dorsal Root Ganglia
Authors: Morikawa Yoshihiro , Tamura Shinobu , Minehata Ken-ichi , Donovan Peter J. , Miyajima Atsushi , Senba Emiko
Published: February 2004

Oncostatin M (OSM) is a member of the interleukin-6 family of cytokines, and we have reported previously that the murine OSM receptor β subunit (OSMR) was expressed in some neurons in the adult trigeminal and dorsal root ganglia (DRGs) and in the perineonatal hypoglossal nucleus. In the present study, we investigated the development of OSMR-positive neurons of DRGs in OSM-deficient mice.In situhybridization revealed that OSMR-positive neurons in DRGs began to appear at postnatal day 0 (P0) and reached the adult level at P14. In the DRGs of the OSM-deficient mice, vanilloid receptor 1 (VR1)- and P2X3-positive small-sized neurons were significantly decreased. In addition, OSMR-positive neurons decreased, resulting in the reduced number of VR1/P2X3/OSMR-triple positive neurons. OSM-deficient mice displayed significantly reduced noxious responses in models of acute thermal, mechanical, chemical, and visceral pain. Thus, OSM plays an essential role in the development of a subtype of nociceptive neurons in the DRGs.
Gating of the cAMP Signaling Cascade and Melatonin Synthesis by the Circadian Clock in Mammalian Retina
Authors: Fukuhara Chiaki , Liu Cuimei , Ivanova Tamara N. , Chan Guy C.-K. , Storm Daniel R. , Iuvone P. Michael , Tosini Gianluca
Published: February 2004

Melatonin is synthesized in retinal photoreceptor cells and acts as a neuromodulator imparting photoperiodic information to the retina. The synthesis of melatonin is controlled by an ocular circadian clock and by light in a finely tuned mechanism that ensures that melatonin is synthesized and acts only at night in darkness. Here we report that the circadian clock gates melatonin synthesis in part by regulating the expression of the type 1 adenylyl cyclase (AC1) and the synthesis of cAMP in photoreceptor cells. This gating is effected through E-box-mediated transcriptional activation of the AC1 gene, which undergoes robust daily fluctuations that persist in constant illumination. The circadian control of the cAMP signaling cascade indicates that the clock has a more general and profound impact on retinal functions than previously thought. In addition, rhythmic control of AC1 expression was observed in other parts of the central circadian axis, the suprachiasmatic nucleus and pineal gland, but not in other brain areas examined. Thus, clock control of the cAMP signaling cascade may play a central role in the integration of circadian signals that control physiology and behavior.
The Number of Glutamate Receptors Opened by Synaptic Stimulation in Single Hippocampal Spines
Authors: Nimchinsky Esther A. , Yasuda Ryohei , Oertner Thomas G. , Svoboda Karel
Published: February 2004

The number of receptors opening after glutamate release is critical for understanding the sources of noise and the dynamic range of synaptic transmission. We imaged [Ca2+] transients mediated by synaptically activated NMDA receptors (NMDA-Rs) in individual spines in rat brain slices. We show that Ca2+influx through single NMDA-Rs can be reliably detected, allowing us to estimate the number of receptors opening after synaptic transmission. This number is small: at the peak of the synaptic response, less than one NMDA-R is open, on average. Therefore, stochastic interactions between transmitter and receptor contribute substantially to synaptic noise, and glutamate occupies a small fraction of receptors. The number of receptors opening did not scale with spine volume, and smaller spines experience larger [Ca2+] transients during synaptic transmission. Our measurements further demonstrate that optical recordings can be used to study single receptors in intact systems.
A Pro-Apoptotic Fragment of the p75 Neurotrophin Receptor Is Expressed in p75NTRExonIVNull Mice
Authors: Paul Christine E. , Vereker Emily , Dickson Kathleen M. , Barker Philip A.
Published: February 2004

The p75 neurotrophin receptor (p75NTR) regulates neuronal survival, apoptosis, and growth. Recent studies have reported that disruption of Exon IV produces a null mouse lacking all p75NTR gene products (p75NTRExonIV-/-), whereas mice lacking p75NTR Exon III (p75NTRExonIII-/-) maintain expression of an alternatively spliced form of p75NTR (s-p75NTR). Here, we report that p75NTRExonIV-/-mice express a p75NTR gene product that encodes a truncated protein containing the extracellular stalk region together with the entire transmembrane and intracellular domains. The gene product is initiated from a cryptic Kozak consensus/initiator ATG sequence within a region of Exon IV located 3′ to the pGK-Neo insertion site. Overexpression of this fragment in heterologous cells results in activation of Jun kinase and induces Pro-caspase-3 cleavage, indicating that it activates p75NTR signaling cascades. These results indicate that aspects of the p75NTRExonIV-/-phenotype may reflect a gain-of-function mutation rather than loss of p75NTR function.
Wahrnehmungsbewusstsein in der Automation – ein neuer „bionischer“ Denkansatz (Perception Awareness in Automation – A New "Bionic" Approach)
Authors: Dietrich Dietmar , Kastner Wolfgang , Schweinzer Herbert
Published: February 2004

Die Kommunikation in der Automatisierung führt zu immer höherer Komplexität, die nicht nur ein Umdenken für die Entwickler notwendig werden lässt, sondern auch grundsätzlich neue Modelle erfordert. Die Kommunikation über Feldbusse ist dabei heute die Voraussetzung, dass Computer Integrated Manufacturing (CIM) wirtschaftlich greifen kann und dass die Gebäudeautomation eine bedeutende Stellung mit Auswirkungen auf die Architektur gewonnen hat. Die Feldbusse sollten deshalb auch eine Schlüsselfunktion in der weiteren Modellentwicklung einnehmen. Von ihnen soll ausgegangen werden. Betrachtet man biologische und psychologische Modelle, erhält man mögliche Antworten, wie solche Modelle in der Automation aussehen könnten. Im vorliegenden Beitrag werden grundlegende Lösungsansätze formuliert, die in gemeinsamen Forschungsaktivitäten mehrerer Institute in den letzten Jahren erarbeitet wurden und in weiterer Folge von reiner Theorie in Praxis umgesetzt werden. Ziel ist ein neuer Ansatz, ein neues hierarchisches Automationsmodell, das Allgemeingültigkeit für viele Bereiche haben kann. Ein entscheidendes Moment ist, dass sich die folgenden Überlegungen von traditionellen Ansätzen lösen, die Vorgehensweise stellt aber im Grunde einen konsequenten Schritt hinsichtlich moderner Forschungsergebnisse dar. Bisher haben sich derartige Ansätze allerdings vor allem auf rein neurologische Überlegungen gestützt, die mit Hilfe der naturwissenschaftlichen Denkweise noch nachzuvollziehen sind, während im Folgenden neueste Ergebnisse der Grundlagenforschung der Wissenschaften der Pädagogik, Psychologie und Psychoanalyse zugrunde gelegt werden, ohne diese jedoch hier breit aufzubereiten.
Ein erweitertes, adaptives Kraftstoffzuteilungskonzept für den Ottomotor (Extended Nonlinear-Adaptive Engine Fuel Supply Control for Spark Ignition Engines)
Authors: Jakubek Stefan , Keuth Nikolaus , Bamer Franz
Published: February 2004

Der Artikel beschreibt zwei neue Methoden zur verbesserten Kraftstoffzuteilung für Verbrennungskraftmaschinen: Die erste eliminiert stationäre Fehler, die z.B. durch ungenaue Kennfelder des volumetrischen Wirkungsgrades oder fehlerhafte Luftmessungen entstehen. Die zweite Methode reduziert instationäre Fehler. Bei Lastsprüngen bildet sich im Ansaugtrakt ein flüssiger Treibstoffwandfilm, was zu einer Abmagerung des Gemisches führt. Um diesen Effekt zu korrigieren, wird eine Zusatzmasse eingespritzt, welche durch ein diskretes Filter berechnet wird. Das Hauptproblem war, einen praxisgerechten und robusten Weg zu finden, um die Filterparameter ausgehend von den verfügbaren λ-Messungen zu adaptieren. Die Leistungsfähigkeit der präsentierten Methoden, gemeinsam mit den entsprechenden Adaptionsalgorithmen, wird anhand von Simulationen mit einem physikalischen Modell eines Ottomotors gezeigt. Die Methoden erreichen in allen Arbeitspunkten des Motors λ-Abweichungen von weniger als einem Prozent.
Berührungsthermometer und ihre Messunsicherheiten in industriellen Anwendungen (Contact Thermometers and their Uncertainties in Industrial Applications)
Authors: Tegeler Erich
Published: February 2004

Temperaturen werden in der überwiegenden Zahl der Fälle mit Thermoelementen oder Widerstandsthermometern gemessen. Anhand von Beispielen werden die wesentlichen in der Praxis auftretenden Schwierigkeiten bei der Temperaturmessung und die damit verbundenen Messunsicherheiten behandelt. Neben der Unsicherheit der Kalibrierung und der Ankopplung des Thermometers an das Messobjekt gehören hierzu auch parasitäre Thermospannungen, verfälschende Leitungswiderstände, Inhomogenitäten von Thermodrähten und verfälschende Strahlungseinflüsse.
Plasma‐based analysis of ERBB2 mutational status by multiplex digital PCR in a large series of patients with metastatic breast cancer
Authors: Corné Julien , Quillien Véronique , Godey Florence , Cherel Mathilde , Cochet Agathe , Le Du Fanny , Robert Lucie , Bourien Héloïse , Brunot Angélique , Crouzet Laurence , Perrin Christophe , Lefeuvre‐Plesse Claudia , Diéras Véronique , De la Motte Rouge Thibault
Published: January 2024

Erb‐b2 receptor tyrosine kinase 2 (ERBB2)‐activating mutations are therapeutically actionable alterations found in various cancers, including metastatic breast cancer (MBC). We developed multiplex digital PCR assays to detect and quantify ERBB2 mutations in circulating tumor DNA from liquid biopsies. We studied the plasma from 272 patients with hormone‐receptor‐positive, human epidermal growth factor receptor 2‐negative (HR+/HER2−) MBC to detect 17 ERBB2 mutations using a screening assay. The assay was developed on the three‐color Crystal dPCR™ naica® platform with a two‐step strategy for precise mutation identification. We found that nine patients (3.3%) harbored at least one ERBB2 mutation. The mutation rate was higher in patients with lobular histology (5.9%) compared to invasive breast carcinoma of no special type (2.6%). A total of 12 mutations were found with the following frequencies: L755S (25.00%), V777L (25.00%), S310Y (16.67%), L869R (16.67%), S310F (8.33%), and D769H (8.33%). Matched tumor samples from six patients identified the same mutations with an 83% concordance rate. In summary, our highly sensitive multiplex digital PCR assays are well suited for plasma‐based monitoring of ERBB2 mutational status in patients with MBC.
Decentralized event‐triggered output feedback adaptive neural network control for a class of MIMO uncertain strict‐feedback nonlinear systems with input saturation
Authors: Bey Oussama , Chemachema Mohamed
Published: January 2024

For a class of multiple‐input multiple‐output large‐scale nonlinear systems in strict‐feedback form with input saturation, external disturbances and immeasurable states, an adaptive decentralized neural network (NN) control strategy on the basis of event triggered mechanism is investigated in this article. In contrast to the literature, the proposed method is centered on the control‐error as a replacement to the tracking‐error that leads to a simplified derivation approach of adaptive laws. Furthermore, the control gains for this class of systems are considered unknown nonlinear functions and not assumed as simple unity or known gains as always done in the literature. Moreover, the challenge of losing controllability that typically arises in state transformation‐based methods, as reported in the literature, is entirely resolved in our approach. Last and not least, all restrictions imposed on unmatched interconnections are eliminated along with avoiding the complexity explosion caused by recursive back‐stepping designs. For this end, the unknown ideal control laws are approximated using NNs, while additional control terms are added to handle saturation effects, unknown interactions, and approximation errors. Additionally, fuzzy inference systems are employed to estimate unknown control errors. Due to the strictly positive real property, the tracking errors are proved to belong to a small compact set using Lyapunov theory. Simulation results demonstrate the effectiveness of the proposed approach.
Electronic Structure Engineering in NiFe Sulfide via A Third Metal Doping as Efficient Bifunctional OER/ORR Electrocatalyst for Rechargeable Zinc‐Air Battery
Authors: Sari Fitri Nur Indah , Lai Yi‐Cheng , Huang Yan‐Jia , Wei Xuan‐Yu , Pourzolfaghar Hamed , Chang Yu‐Hao , Ghufron Muhammad , Li Yuan‐Yao , Su Yen‐Hsun , Clemens Oliver , Ting Jyh‐Ming
Published: January 2024

Ti, V, Cr, Mn, Co, and Cu, have been investigated as a third dopant in NiFe sulfide for enhanced oxygen evolution reaction (OER)/oxygen reduction reaction (ORR). The effects of dopant on surface electronic structure, conductivity, and thermodynamic barrier of reaction are addressed and discussed. For the OER, X‐ray photoelectron spectroscopy analysis shows that electron transferring from the Ni to the dopants enhances the catalytic performance of the sulfide. Cu doped NiFe sulfide exhibits the best OER performance. For the ORR, density functional theory calculation indicates that Ti, V, Mn, Co, and Cu upshift the d‐band center (ɛd), while Cr downshifts the ɛd. Among the dopants, V leads to optimized electronic structure modification, giving optimized adsorption energy of *O on the Ni, the lowest rate determining step ΔG1, and the best ORR activity. By considering E10‐E1/2 together with the maximum current density of the OER and limited diffusion current density of the ORR, NiFeVS exhibits the best OER/ORR bifunctionality. The performance of NiFeVS as a cathodic catalyst has also been evaluated in a zinc air battery, demonstrating a specific capacity of 698 mAh g−1, maximum power density of 190 mW cm−2, and a superior cycle stability of 2400 cycles (400 h).
Enabling High‐Performance Potassium‐Ion Batteries by Manipulating Interfacial Chemistry
Authors: Zhang Haodong , Wang Huwei , Li Wei , Wei Yaojie , Wen Bohua , Zhai Dengyun , Kang Feiyu
Published: January 2024

As a promising candidate for the flame‐retardant electrolyte, triethyl phosphate (TEP)/potassium bis(fluorosulfonyl)amide (KFSI)‐based electrolyte has drawn much attention in the K‐ion battery community. Although the TEP/KFSI formula at a moderate main salt concentration (normally, <3 m) enables the compatibility of the reactive K metal anode, the long‐standing oxidative instability of KFSI salt remains unsolved. Here, an additive strategy is reported to address the high‐voltage issue in the TEP/KFSI electrolyte, and generalize it to the other KFSI‐based electrolytes. The addition of potassium nitrate changes the surface charge distribution and effectively suppresses the decomposition of KFSI toward the high‐voltage cathode. The nitrate‐containing electrolyte enables superior stability of a 4.3 V‐class K‐ion battery, as evidenced by its 80% capacity retention over 2000 cycles (≈6 months) at the 1 C rate. Moreover, the long‐cycling stability of the graphite‐based full cell with Prussian Blue cathode is demonstrated.
2D Conductive Metal–Organic Framework with Anthraquinone Built‐In Active Sites as Cathode for Aqueous Zinc Ion Battery
Authors: Liu Jingjuan , Zhou Yi , Xing Guolong , Qi Meiling , Tang Zhe , Terasaki Osamu , Chen Long
Published: January 2024

2D conductive metal–organic frameworks (2D c‐MOFs) as emerging 2D graphene‐like crystalline materials have become a promising platform for energy storage. However, their capacity is largely constrained by the limited number of electroactive sites. Integrating multiple redox‐active moieties into the 2D c‐MOF skeletons is an efficient strategy toward high‐performance battery cathodes. Herein, by tailoring an anthraquinone‐based multitopic catechol ligand, a novel quinone‐containing copper‐catecholate MOF (Cu‐TBPQ MOF) is successfully developed. The Cu‐TBPQ MOF exhibits abundant porosity, excellent conductivity, and multiple redox‐active sites. These characteristics make it an ideal candidate as a cathode material for zinc ion batteries. Notably, the Cu‐TBPQ MOF demonstrates an impressive reversible specific capacity of 371.2 mAh g−1 at a current density of 50 mA g−1. Furthermore, it exhibits outstanding rate capability and long‐term durability, retaining a capacity of 120.3 mAh g−1 at a high current density of 2.0 A g−1 even after 500 charge–discharge cycles. The successful enrichment of redox‐active sites in the work opens up new avenues for the rational design of electrochemically active 2D c‐MOFs, enhancing their potential for advanced energy storage applications.