bluuebunny/crossref_metadata_2025 huggingface dataset:
Research of protein complex of F3 hybrids chickpea seed
Authors: Molodchenkova O. O. , Lavrova G. D. , Kartuzova T. V. , Bezkrovna L. Ya. , Lykhota O. B. , Ryshchakova O. V. , Levitsky Yu. A.
Published: November 2020

Aim. The aim of the study was to investigate the peculiarities of the protein complex of seeds of F3 hybrid populations and their parental forms of chickpea (Cicer arietinum L.) of Ukrainian and foreign breeding. Methods. Research methods are  standard methodologies of  plant biochemical analysis  and methods developed  in the laboratory (Kjeldahl method, spectrophotometric methods, protein electrophoresis). Results. The peculiarities of protein content, basic fractions of globulins – legumin and vicilin, activity of trypsin inhibitor, lectins, lipoxygenase in the seeds of F3 hybrid populations and their parent forms of chickpea have been established. It was shown using electrophoretic and densitometric analyses that F3 hybrid populations of chickpea  were non identifical  in level and character of  globulin component composition variability.  The clear differences are observed both in the intensity of the staining and the displacement of  protein components of legumin and vicilin which are identical in mobility,  presence and absence of components that are typical for  particular hybrid combination. Conclusions. Obtained results can be used to selection of chickpea genotypes of food direction.

Keywords: chickpea, protein,  vicilin, legumin, trypsin inhibitor.
The Vestigial Heart
Authors: Torras Carme
Published: November 2020

A thirteen-year-old girl wakes up in a future where human emotions are extinct and people rely on personal-assistant robots to navigate daily life.

Imagine a future in which many human emotions are extinct, and “emotional masseuses” try to help people recover those lost sensations. Individuals rely on personal-assistant robots to navigate daily life. Students are taught not to think but to employ search programs. Companies protect their intellectual property by erasing the memory of their employees. And then imagine what it would feel like to be a sweet, smart thirteen-year-old girl from the twenty-first century who wakes from a cryogenically induced sleep into this strange world. This is the compelling story told by Carme Torras in this prize-winning science fiction novel. We meet Celia, brought back to life when a cure is found for her formerly terminal disease, and Lu, Celia's adoptive mother, protective but mystified by her new daughter. There is Leo, a bioengineer, who is developing a “creativity prosthesis” to augment humans' atrophied capacities, and the eccentric robotics mogul Dr. Craft. And there is Silvana, an emotional masseuse who reads old books to research the power of emotion. Silvana sees Celia as a living, breathing example of the emotions and feelings that are now out of reach for most people.

Torras, a prominent roboticist, weaves provocative ethical issues into her story. What kind of robots do we want when robot companions become as common as personal computers are now? Is it the responsibility of researchers to design robots that make the human mind evolve in a certain way? An appendix provides readers with a list of ethics questions raised by the book.
The Joy of Search
Authors: Russell Daniel M.
Published: November 2020

How to be a great online searcher, demonstrated with step-by-step searches for answers to a series of intriguing questions (for example, “Is that plant poisonous?”).

We all know how to look up something online by typing words into a search engine. We do this so often that we have made the most famous search engine a verb: we Google it—“Japan population” or “Nobel Peace Prize” or “poison ivy” or whatever we want to know. But knowing how to Google something doesn't make us search experts; there's much more we can do to access the massive collective knowledge available online. In The Joy of Search, Daniel Russell shows us how to be great online researchers. We don't have to be computer geeks or a scholar searching out obscure facts; we just need to know some basic methods. Russell demonstrates these methods with step-by-step searches for answers to a series of intriguing questions—from “what is the wrong side of a towel?” to “what is the most likely way you will die?” Along the way, readers will discover essential tools for effective online searches—and learn some fascinating facts and interesting stories.

Russell explains how to frame search queries so they will yield information and describes the best ways to use such resources as Google Earth, Google Scholar, Wikipedia, and Wikimedia. He shows when to put search terms in double quotes, how to use the operator (*), why metadata is important, and how to triangulate information from multiple sources. By the end of this engaging journey of discovering, readers will have the definitive answer to why the best online searches involve more than typing a few words into Google.
Repression of interrupted and intact rDNA by the SUMO pathway in Drosophila melanogaster
Authors: Luo Yicheng , Fefelova Elena , Ninova Maria , Chen Yung-Chia Ariel , Aravin Alexei A
Published: November 2020

Ribosomal RNAs (rRNAs) are essential components of the ribosome and are among the most abundant macromolecules in the cell. To ensure high rRNA level, eukaryotic genomes contain dozens to hundreds of rDNA genes, however, only a fraction of the rRNA genes seems to be active, while others are transcriptionally silent. We found that individual rDNA genes have high level of cell-to-cell heterogeneity in their expression inDrosophila melanogaster. Insertion of heterologous sequences into rDNA leads to repression associated with reduced expression in individual cells and decreased number of cells expressing rDNA with insertions. We found that SUMO (Small Ubiquitin-like Modifier) and SUMO ligase Ubc9 are required for efficient repression of interrupted rDNA units and variable expression of intact rDNA. Disruption of the SUMO pathway abolishes discrimination of interrupted and intact rDNAs and removes cell-to-cell heterogeneity leading to uniformly high expression of individual rDNA in single cells. Our results suggest that the SUMO pathway is responsible for both repression of interrupted units and control of intact rDNA expression.
Single-cell multiomic profiling of human lungs reveals cell-type-specific and age-dynamic control of SARS-CoV2 host genes
Authors: Wang Allen , Chiou Joshua , Poirion Olivier B , Buchanan Justin , Valdez Michael J , Verheyden Jamie M , Hou Xiaomeng , Kudtarkar Parul , Narendra Sharvari , Newsome Jacklyn M , Guo Minzhe , Faddah Dina A , Zhang Kai , Young Randee E , Barr Justinn , Sajti Eniko , Misra Ravi , Huyck Heidie , Rogers Lisa , Poole Cory , Whitsett Jeffery A , Pryhuber Gloria , Xu Yan , Gaulton Kyle J , Preissl Sebastian , Sun Xin ,   NHLBI LungMap Consortium
Published: November 2020

Respiratory failure associated with COVID-19 has placed focus on the lungs. Here, we present single-nucleus accessible chromatin profiles of 90,980 nuclei and matched single-nucleus transcriptomes of 46,500 nuclei in non-diseased lungs from donors of ~30 weeks gestation,~3 years and ~30 years. We mapped candidate cis-regulatory elements (cCREs) and linked them to putative target genes. We identified distal cCREs with age-increased activity linked to SARS-CoV-2 host entry gene TMPRSS2 in alveolar type 2 cells, which had immune regulatory signatures and harbored variants associated with respiratory traits. At the 3p21.31 COVID-19 risk locus, a candidate variant overlapped a distal cCRE linked to SLC6A20, a gene expressed in alveolar cells and with known functional association with the SARS-CoV-2 receptor ACE2. Our findings provide insight into regulatory logic underlying genes implicated in COVID-19 in individual lung cell types across age. More broadly, these datasets will facilitate interpretation of risk loci for lung diseases.
Creating Institution-Wide Awareness of, and Engagement with, Open Scholarship
Authors: Colla Eleanor
Published: November 2020

INTRODUCTION Strategies for how, when, and why to communicate on the topics of Open Scholarship (OS) are many and varied. Here, the author reflects on how a small, regional university library went from a low knowledge base of OS to having OS more thoughtfully and thoroughly considered across many aspects of scholarship. DESCRIPTION OF APPROACH The author discusses how, over a three-year period, the library turned OS from being seen as a topic only the library deals with, to a nuanced conversation present across many levels of a university. This was done in three broad stages: first, upskilling librarians; second, reaching out to others working in this space and creating conversations across campus; and third, broadening conversations to different audiences whilst beginning to embed OS in institutional practices. NEXT STEPS Collaborative engagement across various levels has worked well for this university. The library will continue this approach to further embed OS in the culture of the institution while looking to further collaborate across the institution, and working with colleagues and OS advocates in other organizations, groups, and bodies.
Approaches combining methods of Operational Research with Business Process Model and Notation: A systematic review
Authors: Tomaskova Hana , Weber Gerhard-Wilhelm
Published: November 2020

Background

Business process modelling is increasingly used not only by the companies’ management but also by scientists dealing with process models. Process modeling is seldom done without decision-making nodes, which is why operational research methods are increasingly included in the process analyses.

Objective

This systematic literature review aimed to provide a detailed and comprehensive description of the relevant aspects of used operational research techniques in Business Process Model and Notation (BPMN) model.

Methods

The Web Of Science of Clarivate Analytics was searched for 128 studies of that used operation research techniques and business process model and notation, published in English between 1 January 2004 and 18 May 2020. The inclusion criteria were as follows: Use of Operational Research methods in conjunction with the BPMN, and is available in full-text format. Articles were not excluded based on methodological quality. The background information of the included studies, as well as specific information on the used approaches, were extracted.

Results

In this research, thirty-six studies were included and considered. A total of 11 specific methods falling into the field of Operations Research have been identified, and their use in connection with the process model was described.

Conclusion

Operational research methods are a useful complement to BPMN process analysis. It serves not only to analyze the probability of the process, its economic and personnel demands but also for process reengineering.
4D street view: a video-based visualization method
Authors: Kageyama Akira , Sakamoto Naohisa
Published: November 2020

We propose a new visualization method for massive supercomputer simulations. The key idea is to scatter multiple omnidirectional cameras to record the simulation via in situ visualization. After the simulations are complete, researchers can interactively explore the data collection of the recorded videos by navigating along a path in four-dimensional spacetime. We demonstrate the feasibility of this method by applying it to three different fluid and magnetohydrodynamics simulations using up to 1,000 omnidirectional cameras.
Stochastic computing in convolutional neural network implementation: a review
Authors: Lee Yang Yang , Abdul Halim Zaini
Published: November 2020

Stochastic computing (SC) is an alternative computing domain for ubiquitous deterministic computing whereby a single logic gate can perform the arithmetic operation by exploiting the nature of probability math. SC was proposed in the 1960s when binary computing was expensive. However, presently, SC started to regain interest after the widespread of deep learning application, specifically the convolutional neural network (CNN) algorithm due to its practicality in hardware implementation. Although not all computing functions can translate to the SC domain, several useful function blocks related to the CNN algorithm had been proposed and tested by researchers. An evolution of CNN, namely, binarised neural network, had also gained attention in the edge computing due to its compactness and computing efficiency. This study reviews various SC CNN hardware implementation methodologies. Firstly, we review the fundamental concepts of SC and the circuit structure and then compare the advantages and disadvantages amongst different SC methods. Finally, we conclude the overview of SC in CNN and make suggestions for widespread implementation.
Automatic modular design of robot swarms using behavior trees as a control architecture
Authors: Ligot Antoine , Kuckling Jonas , Bozhinoski Darko , Birattari Mauro
Published: November 2020

We investigate the possibilities, challenges, and limitations that arise from the use of behavior trees in the context of the automatic modular design of collective behaviors in swarm robotics. To do so, we introduceMaple, an automatic design method that combines predefined modules—low-level behaviors and conditions—into a behavior tree that encodes the individual behavior of each robot of the swarm. We present three empirical studies based on two missions:aggregationandForaging. To explore the strengths and weaknesses of adopting behavior trees as a control architecture, we compareMaplewithChocolate, a previously proposed automatic design method that uses probabilistic finite state machines instead. In the first study, we assessMaple’s ability to produce control software that crosses the reality gap satisfactorily. In the second study, we investigateMaple’s performance as a function of the design budget, that is, the maximum number of simulation runs that the design process is allowed to perform. In the third study, we explore a number of possible variants ofMaplethat differ in the constraints imposed on the structure of the behavior trees generated. The results of the three studies indicate that, in the context of swarm robotics, behavior trees might be appealing but in many settings do not produce better solutions than finite state machines.
A Reconfigurable Model-Based Design for Rapid Prototyping on FPGA
Authors:   School of Cyber Science and Technology, Beihang University, Beijing, China, Pirzada Syed Jahanzeb Hussain , Murtaza Abid , Xu Tongge , Jianwei Liu
Published: November 2020

The digital design methodologies are evolving with the increase of digital systems utilization in daily life. The Model Based Design (MBD) methodology provides a unique methodology for design and implementation of digital systems on Field Programmable Gate Array (FPGA). Recently, a lot of research effort has been put to exploit new methodologies for designing and prototyping of digital systems on FPGA. The FPGA hardware provides prototyping which provides means of verifying your design at an early stage of development cycle. This helps to evaluate design trade-offs by testing the design in real-time on hardware. Making prototypes is a common practice in research-oriented projects. However, it requires excess development time which increases time to market of the product. This paper illustrates the use of reconfigurable MBD for rapid prototyping of digital systems on Microsemi ACTEL FPGAs for improving the design-cycle and time-to-market of a product. The model is simulated to verify the functionality of the design at system-level and a high-level code is generated from the MBD toolset embedded in MATLAB for hardware implementation. Then, a High-Level Synthesis (HLS) is performed on the generated code which converts this high-level code into Verilog-HDL suitable for hardware implementation on FPGA. Hence, this work presents a methodology and its analysis for design of digital system using high-level synthesis on Microsemi ACTEL FPGA.
Effects of Cadmium Stress on Photosynthetic Physiological Characteristics of Two Rorippa Species
Authors: Ning Yuan Yun- , Yu Wang Lin- , Hao Chen , Feng Xu Yu-
Published: November 2020

The photosynthetic physiological characteristics of Rorippa amphilia and Rorippa sylvestris were studied by pot experiment under Cd stress, and 7 Cd treatments were 0, 1, 5, 10, 50, 100, 200（mg·kg-1）respectively. The results showed that, (1) The Chla, chlb, and Ch(la+b) changed little when the Cd concentration in the soil was less than 100 mg·kg-1. There was no significant difference among the treatments. When Cd concentration was more than or equal to 100 mg·kg-1, all indexes decreased significantly, Chla/b increased gradually with the increase of Cd stressful concentration; (2) Net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) first increased and then decreased with the increase of Cd stressful concentration, while intercellular carbon dioxide concentration (Ci) increased significantly; (3) The photochemical quenching coefficient (qP) and electron transfer rate (ETR)  decreased gradually with the increase of Cd stressful concentration, while the non-photochemical quenching coefficient (NPQ) increased with the increase of Cd stress concentration. It indicated that the photosynthetic mechanism of leaves was damaged; the maximum photochemical quantum yield (Fv/Fm) and potential activity (Fv/Fo) of PSII decreased gradually with the increase of Cd stress concentration. When Cd concentration was more than 50 mg·kg-1, the decrease was very obvious, showing photoinhibition. The concentration of Cd below 50 mg·kg-1 had little effect on photosynthesis. These results provide a theoretical application by using these two species of Rorippa to ecological restore the Cd contaminated farmland and abandoned mines.
A Retrospective Analysis of Copper T380A Intrauterine Device in Rivers State, Nigeria: Side Effects and Discontinuation Rate
Authors: Esther Nonye-Enyidah , Nonyenim Enyidah S. , Jumbo A. I.
Published: November 2020

Background: Intrauterine contraceptive devices (IUCDs) are long term, effective and reversible methods of contraception used worldwide by millions of women. The users discontinue its use despite the effectiveness due to various reasons.

Objective: To determine the prevalence rate, side effects, discontinuation rate and indications for discontinuation of Cu T380A at Rivers State University Teaching Hospital (RSUTH), Port Harcourt.

Methods: A retrospective study of 874 clients attending family planning clinic at the RSUTH from 1st January, 2015 – 31st December, 2019. Their records were retrieved from the clinic and reviewed. Data was extracted, coded and analyzed using the statistical package for social sciences (SPSS) IBM version 25.0 (Armonk, NY).

Results: Of the 874 acceptors of contraceptives, 221 accepted and used Cu T380A within the study period giving a prevalence rate of 25.3%. The mean age was 37.87+ 5.01 years. The modal age group was 35-39 years accounting for 81 (36.7%). Age range was 23-51 years and most (63.8%) were multiparous. Majority of the clients had formal education 217 (98.2%) and were married, 219 (99.1%). Discontinuation rate was 29.4%and the common reasons for discontinuation were menorrhagia, 14 (21.6%); desire for pregnancy, 13 (20%); irregular vaginal bleeding and menopause each contributing 8 (12.3%). Others were missing string, 6 (9.2%), vaginal discharge, sexual problems, lower abdominal and waist pains each accounting for 5 (7.7%) respectively. There was one unintended pregnancy. 

Conclusion: Copper T380A is safe and effective. Menorrhagia and irregular vaginal bleeding were the commonest side effects. Pregnancy desire and side effects were the reasons for its discontinuation.
Mechanism of action of Bacillus thuringiensis var israelensis insecticidal δ‐endotoxin
Authors: Thomas W.E. , Ellar D.J.
Published: July 2002

Bacillus thuringiensis var israelensisδ‐endotoxin protein active against mosquitoes was inactivated by prior incubation with lipids extracted from Aedes albopictus cells. Experiments with lipid dispersions and multilamellar liposomes showed that the toxin binds to phosphatidyl choline, sphingomyelin and phosphatidyl ethanolamine provided these lipids contain unsaturated fatty acids. Phosphatidyl serine binds toxin less efficiently and phosphatidyl inositol, cardiolipin, cerebroside and cholesterol show no affinity for the toxin. The results suggest an insecticidal mechanism in which interaction of toxin with specific plasma membrane lipids causes a detergent‐like rearrangement of the lipids, leading to disruption of membrane integrity and eventual cytolysis.
Neurotensin modulates human neutrophil locomotion and phagocytic capability
Authors: Goldman Rachel , Bar-Shavit Zvi , Romeo Domenico
Published: July 2002

Neurotensin (NT) was found to induce oriented locomotion and augment the phagocytic capability of human blood neutrophils over 10−11–10−7 M. The tridecapeptide also causes Ca2+ extrusion from neutrophils, very likely as a result of intracellular Ca2+ mobilization. It is suggested that the NT‐mediated functional modulation of neutrophils correlates with the capacity of NT to affect the intracellular compartmentalization of Ca2+. Peripheral NT‐elicited phenomena such as vasodilation, enhanced vascular permeability, mast cell degranulation and the enhancement of directional migration and phagocytosis of neutrophils described here, classify NT as a typical mediator of inflammation.
Proteolytic activation and stimulation by Ca2+ of glucan synthase from soybean cells
Authors: Kauss Heinrich , Köhle Harald , Jeblick Wolfgang
Published: July 2002

In homogenates from suspension‐cultured soybean cells, 1,3‐β‐d‐glucan synthase activity is increased through preincubation with trypsin or due to action of an endogenous enzyme which presumably is a protease as it is inhibited by soybean trypsin inhibitor. The 1,3‐β‐d‐glucan synthase in untreated membrane preparations is also reversibly stimulated by Ca2+. This Ca2+‐dependence is lost on proteolytic activation. Regardless as to whether the enzyme was rendered active by preincubation with trypsin or by the presence of Ca2+, it is inhibited by calmidazolium, trifluoperazine and polymyxin B, suggesting that the activation by Ca2+ is not mediated by calmodulin.
The Ca2+sensitivity of the actin‐activated ATPase of scallop heavy meromyosin
Authors: Wells Christine , Bagshaw Clive R.
Published: July 2002

The actin‐activated scallop heavy meromyosin (HMM) ATPase was monitored turbidometrically during a limited number of turnovers. At 4 μM actin, the turnover rate in the presence of Ca2+ (1.2 s−1 per head) was 650‐fold higher than in its absence (1.8 × 10−3 s−1), a Ca2+ sensitivity which approaches that expected in vivo. The extent of Ca2+ activation was much larger than the observed by steady‐state measurements, where the rate, in the absence of Ca2+, is dominated by a small proportion of unregulated molecules. Acto‐HMM formation, and its dissociation by ATP, were Ca2+ insensitive.
Purification and characterization of ubiquitin from mammalian testis
Authors: Loir M. , Caraty A. , Lanneau M. , Menezo Y. , Muh J.P. , Sautiere P.
Published: July 2002

Ubiquitin was extracted from testis of 4 mammals and purified to homogeneity by gel filtration chromatography. Amino acid compositions and NH2‐terminal sequences were found to be identical in the 4 species and with calf thymus ubiquitin. Ubiquitin conformation was shown to be very sensitive to oxidation. Improved methods for radioimmunoassay of ubiquitin in tissue extracts are also discussed.
The cleavable prepiece of an imported mitochondrial protein is sufficient to direct cytosolic dihydrofolate reductase into the mitochondrial matrix
Authors: Hurt Eduard C. , Pesold-Hurt Brigitte , Schatz Gottfried
Published: July 2002

The cleavable prepiece of the precursor to yeast cytochrome c oxidase subunit IV (an imported mitochondrial protein) was attached to the amino‐terminus of mouse dihydrofolate reductase (a cytosolic protein) by gene fusion. The resulting fusion protein was imported into the matrix of isolated, energized yeast mitochondria and cleaved to a polypeptide whose size was similar to that of authentic dihydrofolate reductase.
VIP enhances TRH‐stimulated prolactin secretion of pituitary tumours
Authors: Prysor-Jones R.A. , Silverlight J.J. , Jenkins J.S. , Stevens A.N. , Rodrigues J.L. , Griffiths J.R.
Published: July 2002

Intravenous thyrotrophin releasing hormone (TRH) caused a 6.5‐fold increase in plasma prolactin (PRL) in rats carrying implanted pituitary tumours. Vasoactive intestinal polypeptide (VIP) had no effect, but TRH given after VIP raised TRH stimulated secretion 13‐fold above basal. 31P NMR spectroscopy showed that VIP caused a decrease in high energy metabolites (depleted phosphocreatine, elevated inorganic phosphate and lowered intracellular pH). TRH alone caused a similar but smaller effect; given after VIP, it caused no detectable depletion. We suggest that the changes in high energy metabolite cencentrations reflect increased cellular energy consumption consistent with a priming process (stage 1) in PRL secretion, followed by hormone release (stage 2). VIP induces stage 1 whereas RTH induced both stages.
Synergistic effect of A23187 and a phorbol ester on amylase secretion from rabbit pancreatic acini
Authors: de Pont J.J.H.H.M. , Fleuren-Jakobs A.M.M.
Published: July 2002

The combination of the ionophore A23187 and the phorbol ester 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) stimulates amylase secretion from rabbit pancreatic acini up to a level equal to, or slightly higher than when carbachol is used as stimulant. Each of the two compounds alone gives only a minor stimulation. This synergistic effect of A23187 and TPA supports a role of protein kinase C in pancreatic enzyme secretion.
Fructose 2,6‐bisphosphate and anaerobic metabolism in marine molluscs
Authors: Storey Kenneth B.
Published: July 2002

Fructose‐2,6‐bisphosphate contents of tissues of three species of marine molluscs were measured over a time course of environmental anoxia. During anaerobiosis fructose‐2,6‐P2 levels in soft tissues (mantle, gill, hepatopancreas) decreased rapidly (to levels of 2.5–4% of control in oyster tissues). Content in adductor muscles of bivalves remained constant, however, while fructose‐2,6‐P2 in gastropod foot muscle increased. The drop in fructose‐2,6‐P2 in soft tissues may be part of the overall metabolic depression occurring during anaerobiosis and may function to limit the use of glycolytic carbon for purposes other than energy production.
Evidence that fusicoccin and indole‐3‐acetic acid induce cytosolic acidification of Zea mays cells
Authors: Brummer Benno , Bertl Adam , Potrykus Ingo , Felle Hubert , Parish° Roger W.
Published: July 2002

Auxin
Coleoptile
Cytosolic pH
Fusicoccin
Growth
Membrane potential
Zea mays
External and internal acetylcholinesterase in rat sympathetic neurones in vivo and in vitro
Authors: Goudou Danièle , Verdiere-Sahuque Martine , Rieger François
Published: July 2002

The subcellular distribution of multiple molecular forms of acetylcholinesterase (AChE) in neurones of rat superior cervical ganglion (SCG) was determined both in vivo and in vitro by the use of selective lipidsoluble or ‐insoluble inhibitors. In vivo as well as in vitro, 10 S AChE is mainly outside the cell. In primary cultures of rat SCG neurones, both 4 S and 16 S AChE are mainly inside the cell. In near‐term rat SCG, 4 S and 16 S are more external to the cell than in primary cultures. In adult rat SCG, 4 S AChE is equally distributed inside and outside and 16 S AChE is mainly outside the cell. Thus, specific AChE externalization probably occurs in neuronal cells as a developmentally regulated process.
Enhancement of fibroblast growth factor‐induced diacylglycerol formation and protein kinase C activation by colon tumor‐promoting bile acid in Swiss 3T3 cells
Authors: Takeyama Yoshifumi , Tanimoto Tetsuji , Hoshijima Masahiko , Kaibuchi Kozo , Ohyanagi Harumasa , Saitoh Yoichi , Takai Yoshimi
Published: July 2002

A small amount (50–200 μM) of deoxycholate (DOC), a colon tumor‐promoting bile acid, did not show a direct effect on protein kinase C activity in a cell‐free system, but enhanced fibroblast growth factor (FGF)‐induced diacylglycerol formation and protein kinase C activation in Swiss 3T3 cells. DOC potentiated both reactions induced by submaximal doses of FGF but showed little effect on the maximal levels of the reactions. DOC alone was inactive in eliciting both reactions in the absence of FGF. DOC did not affect the binding of FGF to the cells. Since it has been described that diacylglycerol serves as a messenger for the activation of protein kinase C in the action of FGF in Swiss 3T3 cells [(1985) FEBS Lett. 191, 205‐210], these results suggest that a small amount of DOC increases the sensitivity to FGF of diacylglycerol formation and thereby potentiates protein kinase C activation in this cell line. This action of DOC was in marked contrast to that of 12‐O‐tetradecanoylphorbol‐13‐acetate, a potent tumor‐promoting phorbol ester, which directly activated protein kinase C in cell‐free and intact cell systems.
Incorporation of [3H]glucosamine into keratin‐related polypeptides in pig epidermis
Authors: King Ian A.
Published: July 2002

Metabolic labelling studies have provided evidence for glycosylated keratins in cultured pig epidermis. [3H]Glucosamine was incorporated into five major particulate polypeptides of M
r 68000, 61000, 57000, 53 000 and 48 000. Radioactivity was present in protein‐bound carbohydrate. Non‐enzymic glycation was excluded. Labelling was largely unaffected by tunicamycin indicating that radioactivity was incorporated mainly into O‐linked oligosaccharides. These [3H]glucosamine‐labelled components were closely related to keratins since (i) they had a similar electrophoretic mobility to polypeptides of purified pig prekeratin, (ii) they were immunoprecipitated by anti‐prekeratin serum and (iii) they were incorporated into reconstituted, intermediate‐sized, keratin filaments.
Role of catalase in metabolism of hydrogen peroxide by the perfused rat heart
Authors: Thayer William S.
Published: July 2002

Cardiac metabolism of H2O2 was studied by determining the concentration dependence for H2O2‐stimulated release of GSSG, an indicator for flux through the glutathione peroxidase pathway, in perfused heart preparations. Treatment of rats with aminotriazole in vivo inhibited heart catalase by 83% and shifted the doseresponse curve for GSSG release toward lower H2O2 concentrations. In aminotriazole‐treated rats, 50 μM H2O2 elicited a maximal rate of GSSG release (about 5 nmol per g heart), whereas 200 μM H2O2 was necessary for obtaining a similar rate of GSSG release in control rat hearts. The results show that catalase, although present at low levels of activity in the heart compared to other organs, functions as a major route for detoxication of H2O2 in the myocardium.
Hyaluronic acid in rabbit pericardial fluid and its production by pericardium
Authors: Honda Atsushi , Ohashi Yoshiaki , Mori Yo
Published: July 2002

High‐M
r(> 2 × 106) hyaluronic acid (about 82) was found for the first time in rabbit pericardial fluid. Biosynthetic experiments with minced pericardium from rabbit showed that the high‐M
rhyaluronic acid in the pericardial fluid was actively synthesized by the pericardium from [3H]glucosamine.
Immunoactive chimeric ST‐LT enterotoxins of Escherichia coli generated by in vitro gene fusion
Authors: Sanchez J. , Uhlin B.E. , Grundström T. , Holmgren J. , Hirst T.R.
Published: July 2002

Two different lengths of the gene encoding Escherichia coli heat‐stable toxin (STa) were fused to the carboxy end of the gene coding for the E. coli heat‐labile toxin A‐subunit (LTA). The hybrid genes directed expression of chimeric LTA‐STa proteins. Association of these chimeras with native heat‐labile toxin B‐subunit (LTB) resulted in protein complexes that bound to GM1 ganglioside and thereby could be assayed in a GM1 ELISA. The complexes reacted with monoclonal antibodies against either LTA, LTB or STa indicating that the STa and LT epitopes remained immunologically intact after fusion. Genetically constructed chimeric proteins exhibiting LT and STa antigens on the same molecule may represent a promising approach to development of broadly protective immunoprophylactic agents and/or useful immunodiagnostic reagents for diarrhoeal diseases caused by enterotoxinogenic E. coli.
Localization and quantitation of opsin and transducin mRNAs in bovine retina by in situ hybridization histochemistry
Authors: Brann Mark R. , Young W.Scott
Published: July 2002

Oligodeoxynucleotide probes complementary to a portion of bovine opsin mRNA and transducin mRNA were used for in situ hybridization histochemistry. Within the retina, only photoreceptors expressed mRNAs detectable with these probes, and the majority of both mRNAs were in photoreceptor inner segments. More opsin mRNA was detected than transducin mRNA. In the inner segments 0.54 ± 0.05 copies/μm3 of opsin mRNA and 0.34 ± 0.05 copies/μm3 of transducin mRNA were detected. In the outer nuclear layer, 0.39 ± 0.06 copies/ μm3 of opsin mRNA and 0.27 ± 0.04 copies/μm3 of transducin mRNA were detected.
Deuterium NMR study of head‐group deuterated phosphatidylserine in pure and binary phospholipid bilayers
Authors: Roux Michel , Neumann Jean-Michel
Published: July 2002

Head‐group deuterated 1,2‐dimyristoyl‐sn‐glycero‐3‐phosphorylserine (DMPS) was synthesized. 2H NMR spectra reflect the ionic strength‐dependent polymorphism of DMPS aqueous dispersions. Results obtained with pure DMPS and mixed bilayers with phosphatidylcholine or phosphatidylethanolamine at various NaCl or LiCl concentrations indicate that interactions with Na+ and Li+ have very different effects upon the head‐group quadrupole splittings.
The primary structure of ornithine aminotransferase
Authors: Simmaco Maurizio , John Robert A. , Barra Donatella , Bossa Francesco
Published: July 2002

Tentative assignments of functional residues in rat liver mitochondrial ornithine aminotransferase have recently been made using the amino acid sequence deduced from a cDNA clone [(1985) J.Biol.Chem. 260, 12993‐12997]. Partial sequences obtained using the pure mature protein demonstrate that one of these assignments, that of Lys 292 as the residue that binds the coenzyme pyridoxal phosphate, is correct. However, the identification of the Glu 34‐Gln 35 bond as the site of post‐translational proteolysis is in error. This cleavage occurs instead at Ala 25‐Thr 26.
Metabolic pathways leading to liver glycogen repletion in vivo, studied by GC‐MS and NMR
Authors: Kalderon B. , Gopher A. , Lapidot A.
Published: July 2002

A quantitative analysis of the pathways leading to glycogen repletion in rats was conducted. [U‐13C]Glucose was administered intra‐intestinally into awake fasted animals. The distribution of glucose isotopomers derived from liver glycogen, liver extracts and plasma was performed by GC‐MS and 13C NMR. The potential gluconeogenic precursors for liver glycogen, lactate, alanine, glutamate and glutamine, were also analyzed. The amount of glycogen that is synthesized by the direct pathway was found to be 35%. The 13C enrichment of liver lactate, alanine and glucose is similar, indicating that they are the major precursors for liver glycogen synthesis via the indirect pathway. Our results demonstrate that after 24 h fasting, when glucose is supplied, gluconeogenesis from endogenous sources is not shut off.
Transport of membrane receptors and the mechanics of sexual cell fusion in Chlamydomonas eugametos
Authors: Homan Wieger , Sigon Corrien , van den Briel Wies , Wagter Ron , de Nobel Hans , Mesland Dick , Musgrave Alan , van den Ende Herman
Published: July 2002

During sexual reproduction in the heterothallic, biflagellate, green alga Chlamydomonas, gametes adhere together via their agglutinins, sex‐specific glycoproteins extrinsically bound to the flagellar membrane. Using an antibody specific for a C. eugametos agglutinin, we illustrate that agglutinins engaged in adhesion are transported to the flagellar tips. This tipping phenomenon, together with a particular orientation of the flagella, forms part of the mechanism by which gametes position themselves properly for fusion in pairs.
Evidence that the 116 kDa component of kinesin binds and hydrolyses ATP
Authors: Penningroth Stephen M. , Rose Patricia M. , Peterson Dolores D.
Published: July 2002

Kinesin was prepared from bovine brain as described previously for studies of translocation. A major component of kinesin, (116 kDa) was shown to undergo specific photocrosslinking with [α‐32P]ATP, indicating it was an ATP‐binding polypeptide. A low ATPase activity associated with kinesin was stimulated up to 5‐fold by microtubules to a specific activity of 14 nmol·min−1·mg−1. N‐Ethylmaleimide inhibited both [α‐32P]ATP binding to the 116 kDa polypeptide and microtubule‐stimulated ATPase activity, suggesting that the 116 kDa polypeptide was the catalytic subunit of kinesin. Though the ATPase activity associated with kinesin is low, it may be sufficient to support motility assuming it is coupled to the velocity of translocation.
Activation and desensitization of glycolysis by stimulation of adenylate cyclase in rat reticulocytes
Authors: Kostić M.M. , Maretzki D. , Źivković R. , Krause E.G. , Rapoport S.M.
Published: July 2002

Isoproterenol or forskolin induce a 10–15‐fold increase in concentration of cyclic AMP in rat reticulocytes as compared with the basal level of 2.3 ± 0.3 μM. Glycolysis is stimulated by both compounds transiently more than 2‐fold with a peak after 7.5 min followed by an exponential decline. The glycolytic rate in the presence of 1O μM isoproterenol or 10 μM forskolin did not return to basal levels within 60 min of incubation, but was depressed by as much as 50% under the influence of 100 μM forskolin. This phenomenon is designated as metabolic desensitization. The stimulation of glycolysis is probably due to activation of phosphofructokinase as well as to stimulation of Na+,K+‐ATPase. The diminished glycolytic flux during the period of metabolic desensitization is accompanied by a decline of glucose 6‐phosphate and in the presence of high concentrations of forskolin also by a decrease in glucose 1,6‐bisphosphate. A lower rate of influx of glucose is postulated.
Location and sequence characterization of the major phosphorylation sites of the high molecular mass neurofilament proteins M and H
Authors: Geisler Norbert , Vandekerckhove Joel , Weber Klaus
Published: July 2002

Diagonal fingerprinting allows the specific purification of those tryptic peptides which change electrophoretic mobility due to a dephosphorylation step introduced after the first dimension. Nine tryptic peptides from the tail domain of porcine neurofilament M protein identify a minimum of 6 phosphorylated serines. Unexpectedly, four of the nine peptides characterize a region of degenerate repetitive sequences. Results on neurofilament H tail, although less complete, yield longer sequences of degenerate repetitive character. Here, all serines present appear to be contained in a lysine‐serine‐proline unit. This motif also occurs in some but not all M peptides. We suggest that degenerate repetitive sequences in neurofilament M and H tails have a high species‐specific drift.
Removal of 33 kDa extrinsic protein specifically stabilizes the S2QA− charge pair in photosystem II
Authors: Vass Imre , Ono Taka-aki , Inoue Yorinao
Published: July 2002

Removal of the 33 kDa extrinsic protein from photosystem (PS) II membranes resulted in markedly increased stabilization of the S2QA
− charge pair as measured by thermoluminescence. The stabilization increase was specific for the S2QA
− charge pair and did not require any special herbicide. The effect was fully reversed by reconstitution with the 33 kDa protein, but not at all by high concentrations of Cl−, as opposed to those effects known to be reversed by 200 mM Cl− . The data are interpreted as indicating a structural change of the donor and/or acceptor side of PS II dependent on association with the 33 kDa extrinsic protein.
A novel quinone: Quinone methide isomerase generates quinone methides in insect cuticle
Authors: Saul Steven , Sugumaran Manickam
Published: July 2002

Insoluble cuticle‐bound enzyme(s) of Manduca sexta pharate pupae, which is known to convert N‐acetyldopamine to N‐acetylnorepinephrine through the intermediate formation of quinone methide, also converted exogenously supplied N‐acetyldopamine quinone to N‐acetylnorepinephrine. The presence of a quinone trap such as N‐acetylcysteine in the reaction mixture containing N‐acetyldopamine and cuticle prevented the formation of N‐acetylnorepinephrine but readily yielded N‐acetylcysteine‐N‐acetyldopamine quinone adduct as a dead‐end product. These results indicate the oxidation of N‐acetyldopamine to its quinone and its enzyme‐catalyzed isomerization to quinone methide before yielding N‐acetyl‐norepinephrine as the stable product. The role of this newly discovered isomerase in sclerotization of insect cuticle is discussed.
Different tachykinin receptor subtypes are coupled to the phosphoinositide or cyclic AMP signal transduction pathways in rat submandibular cells
Authors: Laniyonu A. , Sliwinski-Lis E. , Fleming N.
Published: July 2002

Tachykinins of different classes (NK1, NK2, NK3) caused the concentration‐dependent synthesis of IP3 in rat submandibular acinar cells with the potency rank order of NK1 < NK2 > NK3. Enhancement of IP3 was not affected by pertussis toxin treatment. The reverse rank order was found in the tachykinin inhibition of isoproterenol‐induced cAMP synthesis and this inhibition was abolished by pertussis toxin, an inactivator of the adenylate cyclase Gi regulatory protein. It is suggested that different tachykinin receptor subtypes are preferentially coupled to phospholipase C or adenylate cyclase by separate G regulatory proteins in rat submandibular acinar cells.
Proteolytic signal sequences (PEST) in the mammalian aminoacyl‐tRNA synthetase complex
Authors: Jacobo-Molina Alfredo , Villa-Garcia Manuel , Chen Hao-Chia , Yang David C.H.
Published: July 2002

Eight aminoacyl‐tRNA synthetases together with three unidentified proteins are associated as a multi‐enzyme complex in mammalian cells. Partial peptide sequences for lysyl‐ and aspartyl‐tRNA synthetases are determined and no highly hydrophobic peptides are found. The partial amino acid sequences for two of the unidentified proteins in the complex are shown to have substantial homology and each has a number of unique sequences. The results suggest that the two unidentified proteins are fragments of synthetases. The partial sequences revealed the presence of PEST sequences in at least three proteins. Inasmuch as PEST sequences are signals for intracellular degradation, the mammalian synthetase complex may have evolved to protect these synthetases against intracellular proteolysis.
Molecular cloning and sequencing of the chicken smooth muscle myosin regulatory light chain
Authors: Messer Neil G. , Kendrick-Jones John
Published: July 2002

A cDNA probe was constructed from a chicken skeletal muscle regulatory light chain cDNA and was used to screen a chicken gizzard cDNA library. A clone containing the entire coding region of the chicken gizzard regulatory light chain was isolated and sequenced. The deduced protein sequence is identical to the most recently reported chemical sequence of the chicken smooth muscle regulatory light chain, and has homologies with other troponin C‐like calcium‐binding proteins.
Translocation of protein kinase C in porcine thyroid cells following exposure to thyrotropin
Authors: Ginsberg Jody , Murray Patricia G. , Parente Janice E. , Wong Kenneth
Published: July 2002

We have previously shown that protein kinase C activators modulate differentiated thyroid function in vitro; however, how protein kinase C may be activated physiologically is unknown. The present studies were undertaken in order to determine whether TSH could activate protein kinase C in vitro. Following exposure of porcine thyroid cells to TSH, translocation of protein kinase C from the cytosol to its membrane‐bound form was observed. Maximal translocation occurred at the lowest TSH concentration able to trigger this response (10 mU/ml) but persisted at higher concentrations (20–100 mU/ml). Time‐course studies revealed that translocation of protein kinase C was seen only after 40 min. TSH could also produce a similar translocation in human neutrophils (known to have TSH receptors). In thyroid cells pre‐treated with TSH, modulation of phorbol‐mediated protein kinase C translocation was noted. These results indicate that TSH causes the translocation of protein kinase C in porcine thyroid cells (and possibly other TSH receptor‐containing cells) and therefore may regulate the action of protein kinase C on differentiated thyroid function.
Amino acid substitutions in mitochondrial ATP synthase subunit 9 of Saccharomyces cerevisiae leading to venturicidin or ossamycin resistance
Authors: Galanis Maria , Mattoon James R. , Nagley Phillip
Published: July 2002

A series of mitochondrially inherited mutants of yeast has been analysed, which were previously identified as showing resistance to the antibiotics venturicidin or ossamycin and whose mutations showed tight linkage to oligomycin‐resistance alleles affecting subunit 9 of the mitochondrial ATP synthase. DNA sequence analysis of the oli1 gene of these mutants has been used to define the nature of amino acid substitution in the subunit 9 protein. In the case of the two venturicidin‐resistant mutants, mutations affect amino acids on the N‐terminal stem of the protein, namely G1y25 → Ser (venR ossS oliR) and Ala 27 → Gly (venR ossS oliS). The mutations found in the two ossamyein‐resistant mutants affect amino acids on the C‐terminal stem of the protein, namely Leu53 → Phe (vanS ossR oliR) and Leu57 → Phe (venS OSSR oliS). These results allow us to further develop a fine structure map of domains within the subunit 9 protein involved in antibiotic interaction.
Identification of 1,4‐thiomorpholine‐3‐carboxylic acid (TMA) in normal human urine
Authors: Matarese R.M. , Solinas S.P. , Montefoschi G. , Ricci G. , Cavallini D.
Published: July 2002

The sulfur‐containing cyclic imino acid 1,4‐thiomorpholine‐3‐carboxylic acid (TMA) has been identified in normal human urine. After the enrichment procedure with ion‐exchange chromatography, the urine extracts were reacted with diazomethane. Gas‐liquid chromatography revealed the presence of two peaks with the same retention times exhibited by authentic TMA after the same derivatization. The two compounds have been identified by mass‐spectrometry as the monomethylated and dimethylated derivatives of TMA. This result represents the first indication of the occurrence of TMA in a mammalian sample.
Identification of photosystem I components from the cyanobacterium, Synechococcus vulcanus by N‐terminal sequencing
Authors: Koike Hiroyuki , Ikeuchi Masahiko , Hiyama Tetsuo , Inoue Yorinao
Published: July 2002

The photosystem I core complex isolated from a thermophilic cyanobacterium, Synechococcus vulcanus, is composed of eight low‐molecular‐mass proteins of 18, 14, 12, 9.5, 9, 6.5, 5 and 4.1 kDa in addition to the PS I chlorophyll protein. N‐terminal amino acid sequences of all these components were determined and compared with those of higher plants. Clearly, the 9.5 kDa component corresponds to the protein which carries the non‐heme iron‐sulfur centers A and B. This protein is so poorly visualized by staining that it has probably been overlooked in gel electrophoresis analyses. The 18, 14, 12 and 9 kDa components show appreciable homology with respective subunits of higher plant PS I. In contrast, the 6.5, 5 and 4.1 kDa components do not correspond to any known proteins except that the sequence of the 4.1 kDa component matches an unidentified open reading frame (ORF) 42 (liverwort) or ORF44 (tobacco) of chloroplast DNA.