bluuebunny/crossref_metadata_2025 huggingface dataset:
HR 4.0 in Manufacturing: Using AI and Big Data to Enhance Workforce Agility and Reduce High Employee Turnover Intentions
Authors: Abrar Fatima , Ahmed Rameez , Khoso Ikhtiar Ahmed , Farooq Nadeem , Ibrahimi Ikramullah
Published: February 2025

The human resource management field in manufacturing operations experienced major transformations because of Industrial Revolution 4.0 (IR 4.0) with HR 4.0 as its main advancement combining artificial intelligence systems with big data analytics. The merged system has built organizational employee flexibility and automated HR procedures leading to reduced employee departures. Through the integration of AI in HRM organizations have gained the ability to analyze advanced predictive data alongside improved employee engagement and optimized talent control which has boosted their HR process automation capability. The research evaluates how the implementation of AI algorithms for HR management impacts employee retention outcomes through big data examination. Secondary statistical research of previously collected data examines the part AI technology plays in enhancing employee retention and minimizing staff departures. The paper applies predictive models based on AI algorithms and big data which predict turnover risks. The predictive models assist organizations in detecting risky staff members and facilitate early prevention plans. To evaluate the relationship between AI-powered HR practices and turnover intentions the researchers have applied correlation methods together with regression analysis of Variance (ANOVA) and chi-square testing. The research employs statistical analysis to assess how AI-based solutions impact employee satisfaction levels and efficiency as well as decrease employee turnover rates. The implementation of AI and big data analytics succeeds in both launching employee retention programs effectively and increasing organizational efficiency through improved HR management better decision systems and adaptable workforce models that match Industry 4.0 manufacturing sector requirements. Organizations use these methods to create highly engaged and efficient staff who maintain stability which leads to better productivity and decreased expenses from employee replacement and new hire recruitment.
A eclesiologia fundamental de Lutero para o diálogo ecumênico atual
Authors: Toné Arthur Jordan de Azevedo
Published: February 2025

A unidade eventual entre católicos e luteranos somente terá algum significado se houver uma compreensão cabal das dificuldades existentes no tempo da Reforma originadas do contexto interpretativo da atividade de Martinho Lutero. Esta pesquisa pretendeu identificar os pontos de convergência entre o pensamento eclesiológico de Lutero e o ensinamento da Lumen gentium do Concílio Vaticano II. Para isto, analisou-se a natureza sacramental da Igreja, identificando o limite de um pluralismo teológico verdadeiro, e cinco das principais obras eclesiológicas de Lutero. Foram identificados oito pontos de consenso entre a eclesiologia fundamental católica e a de Lutero, alguns dos quais não tratados suficientemente nos diálogos ecumênicos oficiais. Fundando-se neles, foram sugeridas nove propostas práticas de comunhão concreta. A caminhada ecumênica poderá se beneficiar com esta pesquisa: (1) porque ela elenca propostas para transformar em experiência prática de formação e vivência cristã os consensos obtidos; (2) porque ela serve para purificar a memória dos cristãos católicos a respeito do pensamento de Lutero sobre esse tema.
What happens when grit goes bad? Conceptualizing “dark” grit among sport employees
Authors: Oja Brent , Jordan Samantha
Published: February 2025

This paper details the complexities of the construct known as “grit” and specifically examines the potential for the positive effects of grit to turn maladaptive or “dark.” Although grit is commonly viewed as a generative construct that helps individuals achieve success and experience well-being, past scholars have suggested there could be a “too much of a good thing” effect whereby the helpful outcomes of grit reach a turning point and become detrimental. We propose that the sport industry is an ideal environment for this phenomenon to occur because the industry has high labor demands, and its employees have a strong passion for sport. When sport employees over engage with their work, they are disposed to the negative outcomes of dark grit, which we propose as burnout and turnover. The paper concludes with an overview of the possible future directions for the study of dark grit among sport employees.
A Conceptual Framework of Work Passion and Key Intelligences among Coaches
Authors: Doh Paul , Kim Minjung , Oja Brent
Published: February 2025

This study proposes a conceptual framework that integrates work passion and three key intelligences: cultural intelligence, digital intelligence, and emotional intelligence, to enhance coaching effectiveness and psychological well-being in sport. The framework explores the anticipated antecedents and outcomes of work passion and these intelligences among coaches. The proposed framework contributes theoretically by highlighting the role of work passion and integrative intelligences as foundational elements for innovative work behaviors in sport coaching, which ultimately enhancing both coaching effectiveness and psychological well-being among coaches. This framework also offers practical insights for sport organizations to strategically invest in coaching development, promoting adaptive, tech-savvy, and inclusive coaching strategies that meet the demands of today’s dynamic sport environment.
Perspectives on Passion in Sport Management Research
Authors: Magnusen Marshall
Published: February 2025

The focus of this special issue is to continue to broaden discussions of passion in sports to management contexts with sport personnel rather than marketing contexts with sport consumers. This is done through an offering of several “think pieces” with the goal of inspiring new and creative scholarship about passion in sport work environments.
Rectification of Dust Particles in a Dusty Plasma Metal Straight Ratchet
Authors: ZHANG Shunxin  , WANG Shuo  , LIU Xue  , WANG Xinzhan  , LIU Fucheng  , HE Yafeng  ,   1 College of Physics Science and Technology, Hebei University, Baoding 071002;,   2 Hebei Research Center of the Basic Discipline for Computational Physics, Baoding 071002;,   3 Hebei Key Laboratory of High-precision Computation and Application of Quantum Field Theory, Baoding 071002
Published: February 2025

Utilizing the principle of the Feynman ratchet, it is possible to rectify the random motion into directed flow of particles under a nonequilibrium environment. In this paper, an experimental setup for a dusty plasma metal straight ratchet is designed to create an asymmetric plasma environment along the ratchet channel, enabling a controllable rectification of micron-sized dust particles. Monodispersed dust particles can form a directional flow in the ratchet channel, and the transport direction could be precisely controlled by adjusting the discharge power and the gas pressure. Research on the transport of dust particles of varying sizes proves that the rectification effect is universal. To reveal the rectification mechanism of dust particles, a fluid model of plasma is employed to calculate the two-dimensional distribution of plasma parameters within the ratchet channel. Further research through Langevin simulation shows that dust particles experience ratchet potentials with distinct asymmetric orientations at different suspension heights within the ratchet channel, leading to different transport directions. The results of this paper provide a theoretical and experimental foundation for further achieving the separation of bi-disperse particles in dusty plasma metal straight ratchets.
Mapping kinase domain resistance mechanisms for the MET receptor tyrosine kinase via deep mutational scanning
Authors: Estevam Gabriella O , Linossi Edmond , Rao Jingyou , Macdonald Christian B , Ravikumar Ashraya , Chrispens Karson M , Capra John A , Coyote-Maestas Willow , Pimentel Harold , Collisson Eric A , Jura Natalia , Fraser James S
Published: February 2025

Mutations in the kinase and juxtamembrane domains of the MET Receptor Tyrosine Kinase are responsible for oncogenesis in various cancers and can drive resistance to MET-directed treatments. Determining the most effective inhibitor for each mutational profile is a major challenge for MET-driven cancer treatment in precision medicine. Here, we used a deep mutational scan (DMS) of ~5764 MET kinase domain variants to profile the growth of each mutation against a panel of 11 inhibitors that are reported to target the MET kinase domain. We validate previously identified resistance mutations, pinpoint common resistance sites across type I, type II, and type I ½ inhibitors, unveil unique resistance and sensitizing mutations for each inhibitor, and verify non-cross-resistant sensitivities for type I and type II inhibitor pairs. We augment a protein language model with biophysical and chemical features to improve the predictive performance for inhibitor-treated datasets. Together, our study demonstrates a pooled experimental pipeline for identifying resistance mutations, provides a reference dictionary for mutations that are sensitized to specific therapies, and offers insights for future drug development.
Negative regulation of miRNAs sorting in EVs: the RNA-binding protein PCBP2 impairs SYNCRIP-mediated miRNAs EVs loading
Authors: Marocco Francesco , Garbo Sabrina , Montaldo Claudia , Colantoni Alessio , Quattrocchi Luca , Gaboardi Gioele , Cicchini Carla , Tartaglia Gian Gaetano , Battistelli Cecilia , Tripodi Marco
Published: February 2025

While it is accepted that Extracellular Vesicles (EVs)-mediated transfer of microRNAs contributes to intercellular communication, the knowledge about molecular mechanisms controlling the selective and dynamic miRNA-loading in EVs is still limited to few specific RNA-binding proteins interacting with sequence determinants. Moreover, although mutagenesis analysis demonstrated the presence/function of specific intracellular retention motifs, the interacting protein/s remained unknown. Here, PCBP2 was identified as a direct interactor of an intracellular retention motif: RIP coupled to RNA pull down and proteomic analysis demonstrated that it binds to miRNAs embedding this motif and mutagenesis proved the binding specificity. Notably, PCBP2 binding requires SYNCRIP, a previously characterized miRNA EV-loader as indicated by SYNCRIP knock-down. SYNCRIP and PCBP2 may contemporarily bind to miRNAs as demonstrated by EMSA assays and PCBP2 knock-down causes EV-loading of intracellular microRNAs. This evidence highlights that multiple proteins/miRNA interactions govern miRNA compartmentalization and identifies PCBP2 as a dominant inhibitor of SYNCRIP function.
An improved termite life cycle optimizer algorithm for global function optimization
Authors: Wang Yanjiao , Wei Mengjiao
Published: February 2025

The termite life cycle optimizer algorithm (TLCO) is a new bionic meta-heuristic algorithm that emulates the natural behavior of termites in their natural habitat. This work presents an improved TLCO (ITLCO) to increase the speed and accuracy of convergence. A novel strategy for worker generation is established to enhance communication between individuals in the worker population and termite population. This strategy would prevent the original worker generation strategy from effectively balancing algorithm convergence and population diversity to reduce the risk of the algorithm in reaching a local optimum. A novel soldier generation strategy is proposed, which incorporates a step factor that adheres to the principles of evolution to further enhance the algorithm’s convergence speed. Furthermore, a novel replacement update mechanism is executed when the new individual is of lower quality than the original individual. This mechanism ensures a balance between the convergence of the algorithm and the diversity of the population. The findings from CEC2013, CEC2019, and CEC2020 test sets indicate that ITLCO exhibits notable benefits regarding convergence speed, accuracy, and stability in comparison with the basic TLCO algorithm and the four most exceptional meta-heuristic algorithms thus far.
Identification of potential insect ecological interactions using a metabarcoding approach
Authors: Borsato Nicole D. , Lunn Katherine , Garrett Nina R. , Biganzoli-Rangel Alejandro José , Marquina Daniel , Steinke Dirk , Floyd Robin , Clare Elizabeth L.
Published: February 2025

Species interactions are challenging to quantify, particularly when they happen cryptically. Molecular methods have become a key tool to uncover these interactions when they leave behind a DNA trace from the interacting organism (e.g., pollen on a bee) or when the taxa are still present but morphologically challenging to identify (e.g., microbial or fungal interactions). The decreasing costs of sequencing makes the mass analysis of thousands of target species possible. However, the challenge has shifted to selecting molecular markers which maximize information recovery while analyzing these data at broad biological scales. In this manuscript we use model arthropod groups to compare molecular markers and their analysis across life stages. We develop protocols for two ecologically and economically devastating pests, the spongy moth (Lymantria dispar dispar) and the emerald ash borer (Agrilus planipennis), and a group of pollinators including bees and wasps which regularly deposit eggs in “bee hotels” where the larvae develop. Using Illumina MiSeq and Oxford Nanopore MinION platforms we evaluate seven primer pairs for five molecular markers which target plants, fungi, microbes, insects, and parasitic phyla (e.g., nematodes). Our data reveals hundreds of potential ecological interactions and establishes generalized methods which can be applied across arthropod host taxa with recommendations on the appropriate markers in different systems. However, we also discuss the challenge of differentiating co-occurring DNA signals and true ecological interactions, a problem only starting to be recognized as eDNA from the environment accumulates on living organisms.
Macroeconomic Factors and Stock Market Performance Based on New Evidence: A Case Study from Nigeria
Authors: Akanbi Abiodun
Published: February 2025

The nexus between macroeconomic conditions and stock market performance is multifaceted in nature. One of the considerations of investors before investing in the stock market is to assess the macro-economic environment of the country. There have been divergent views on the relationship between stock market and macro-economic variables in literature and this debate is inconclusive. This study included the variable that measures the overall macroeconomic environment i.e. Country Policy and Institutional Assessment (CPIA), which is imperative and novel because it has been ignored in the past. This study provided empirical evidence on the relationship between selected macro-economic factors and stock market performance in Nigeria from 2003 to 2022. The foundational theory for this study is the Arbitrage Pricing Theory (APT). The study employed the Autoregressive Distributed Lag (ARDL) cointegration approach to examine relationship between macroeconomic variables and stock market performance in Nigeria. The ARDL results established a short-run linear relationship and non-existence of long run relationship among the variables. The results suggested that stock performance in Nigeria was influenced positively by GDP growth although it was an insignificant influence. However, the study that the found overall macroeconomic environment, inflation, and interest rate spread, having a negative yet insignificant influence over the stock market performance. These findings are important for policy makers because the overall macro-economic environment considers the effectiveness of the policies and actions taken as well as the institutional arrangements a country's government and central bank to regulate and control the economy. This study recommends the need for proper institutional arrangement and policy framework to manage and maintain stable macro-economic environment to promote stock market performance.
Sustainable Libraries for Books and Bytes: A Bibliometric Study of Web of Science from 2005-2024
Authors: Sahu Lili , Maharana Bulu
Published: February 2025

A library is termed sustainable when it's planned and operated, reducing environmental impact as far as possible. Sustainable libraries, therefore, consider all three aspects of sustainability: social, economic, and environmental. The concept of sustainable libraries, which integrate traditional books and digital resources to adapt to technological and environmental changes. This paper emphasizes the importance of sustainable practices in library management, including eco-friendly building designs, digital collections, and waste reduction. The paper also delves into a bibliometric study on sustainable libraries, analysing academic literature to identify trends, influential contributions, and research growth patterns. For the bibliometric analysis and visualization VOSviewer software is used based on 248 literature data on sustainable libraries papers gathered from the Web of Science core collection from 2005 to 2024 on the subject discipline of Library and Information Science. The year 2021 has the highest number of records, with 25 entries, and the most prolific author was found to be Henrik Jochumsen, with a total link strength of 32.  This study also provides insights into the global distribution of research on sustainable libraries, highlighting key countries, document types, and open access modes.
Spatial Variability and Pollution Loads of Selected Heavy Metals in Cocoa Plantation Soils in Cross River State, Nigeria
Authors: Olim Denis Michael , Nwachukwu Olayinka Ibiwumi , Onwuka Mabel Ifeoma
Published: February 2025

Understanding heavy metal concentrations and their spatial variability is crucial for improved soil health and increased crop productivity. This study assessed heavy metal concentrations and their pollution loads in soils of cocoa plantations in Ikom, Etung and Boki Local Government Areas of Cross River State. Sixty (60) composite soil samples were collected from the area and analyzed Cd, Cr, Pb and Al. The results obtained revealed that Cd was above the maximum permissible limit for soil. Lead had means of 0.011 mg/kg, 0.02 mg/kg and 0.010 mg/kg, Cd had means of 1.53 mg/kg, 1.45 mg/kg and 1.25 mg/kg while Cr was 0.05 mg/kg, 0.03 mg/kg and 0.02 mg/kg Ikom, Etung and Boki respectively. The contamination factor for Cr and Pb was low (CF < 1) while Cd was considerably contaminated (3≤ CF < 6). The results of PLI indicated an unpolluted condition (PLI<1). Apart from Cd, the values of Igeo for Cr, Pb and Al fell in class ‘0’, indicating practically uncontaminated conditions in these areas. The soils of the area are safe for crop production but measures should be taken to reverse the trend of high concentration of Cd in the soils.
Effect of Mexican Sunflower (Tithonia diversifolia) Extract on the Growth and Herbage Yield of Lemon Grass (Cymbopogon flexuosus) var. Krishna
Authors: Balaji K. Ayyan Arvind , Venkatesan S.
Published: February 2025

A field experiment was conducted in the Medicinal plant unit, Department of Horticulture, Faculty of Agriculture, Annamalai university to study the effect of mexcian sunflower (Tithonia diversifolia) leaf extract on the growth and herbage yield of lemon grass (Cymbopogon flexuosus var. krishna). The experiment was laid out in a Randomized Block Design with nine treatments and replication thrice. The treatments consisted of FYM (25 t ha-1) and Tithonia extract at different concentration viz.,5, 10, 15, 20, 25, 30, 35 and 40 per cent as foliar application on 15, 45, 75, 105, and 135 days after transplanting. The results of the experiment revealed that the combined application of FYM 25 t ha-1 + Tithonia extract per cent 40 per cent (T9) produced the highest values for growth parameters such as plant height, number of tillers, plant spread (East-West and North-South), number of leaves plant-1 and yield parameters such as fresh herbage, dry herbage yield and dry matter production (per plant, per plot and per hectare).
Green Textile Innovation: Silk Dyeing with Temple Floral Waste
Authors: Mishra Vaishali , Verma Meenu , Singh Akanksha , Singh K.J. , Tewari S.K. , Pal Mahesh
Published: February 2025

The disposal of temple floral waste, including marigold (Tagetes erecta L.) flowers, into water bodies contributes significantly to river pollution. This study presents a novel and eco-friendly approach for utilizing marigold floral waste as a natural dye for silk fabrics, eliminating the need for chemical solvents in the dyeing process. The aqueous extraction method was employed to obtain the dye, highlighting its sustainability and alignment with green chemistry principles. The dyeing process was conducted on 100% silk fabrics under optimized conditions (30 to 80°C). The dyed fabrics exhibited vibrant and uniform coloration, which was analyzed using a Colorimeter to determine L*, a*, and b* values. The results demonstrated high color fastness, with the surface color remaining intact even after a 30-minute washing cycle at 60°C. This is the first study to successfully utilize marigold floral waste for silk dyeing without the use of chemical solvents, offering a sustainable alternative to synthetic dyes. By repurposing temple waste and avoiding harmful chemicals, this method addresses environmental concerns associated with both floral waste disposal and textile dye pollution. The findings highlight the potential of this innovative approach for sustainable textile coloration while contributing to waste management and pollution reduction.
Extension Advisory Services on Farmer’s Climate Change Mitigation Strategies in Misungwi District, Tanzania: Issues and Options
Authors: Samweli Sagire Nelson , Martin R. , Ntumva M.
Published: February 2025

This study evaluates the impact of extension advisory services on farmers’ climate change mitigation strategies. Questionnaires were completed by 64 extension agents and data were analysed using descriptive statistics. The study found that the most common method used for identifying farmers’ climate change adaptation needs is extension agents visiting farmers (84.6%), followed by training sessions are less frequently used (26.2%), and other methods are rarely employed (1.5%). Insufficient training on climate change, with a frequency of 55, resource constraints, reported by around 49 respondents and limited access to climate information also stand out as a significant issue, with a frequency of about 41, and lastly knowledge barriers with 39 respondents were shown as the challenges encountered during dissemination of advisory services to farmers. This study revealed that majority (56.9%) of extension agents reported receiving transport facilities, while 41.5% had access to timely updated information. Though, only 18.5% of respondents indicated receiving regular training. This study concludes that the majority of extension agents have reported that they do not receive sufficient support in their efforts to educate farmers on how to adapt to climate change. The extension agents are not prepared to deal with the complexities of climate change, despite the fact that they rely on personal visits to capture the needs of the farmers. The low level of diversity engagement approaches, the low rates of hands-on training, and the moderate perceptions of comfort in communicating climate information are all indicators that this is the case. In addition, many extension agents believe that the increase of these concerns is caused by resource problems, which include inadequate access to timely information, inadequate training, and inadequate financial resources. This study recommends that the government should allocate enough resources aimed at supporting the extension agents in fulfilling their operations.
Microbial Contamination of Smoked Tilapia Fish Sold in Kure Market, Minna, Niger State, Nigeria
Authors: H. Kanki, , A.O. Omotosho, , S. Adamu, , U. Okekeaji, , M. S. Akor, , M. M. Lemu,
Published: February 2025

This study assessed the microbial load and diversity in smoked tilapia fish sold in Kure Market, Minna, Niger State, Nigeria. A total of 40 samples were analyzed using standard microbiological methods. The results revealed a mean bacterial colony count ranging from 120 ± 13.03 to 189 ± 11.03 CFU/ml across different parts of the fish. Seven bacterial species, including pathogenic strains such as Salmonella sp. and Escherichia coli, were isolated alongside four fungal species, including Aspergillus flavus, a known producer of aflatoxins. The presence of these microorganisms poses serious public health risks, highlighting the urgent need for improved fish handling, processing, and storage practices to safeguard consumer health. These findings underscore the need for improved handling, processing, and storage practices to ensure the safety of smoked fish and protect consumer health.
Integrated Farming Systems Effect on Farmers Livelihood Security in Uttar Pradesh, India
Authors: Mishra Avinash , Thapa Twinkle , Baboo Akash , Yadav Randhir , Laishram Rabina
Published: February 2025

The aim of the present investigation was to determine the degree of livelihood security experienced by farmers in the Sultanpur region of Uttar Pradesh. Kurebhar and Dhanpatganj blocks were selected based on having the maximum acreage under an Integrated Farming System. Both primary and secondary data were used in the investigation. To achieve the objectives of the study, primary data were collected from 150 randomly selected farmers for the years 2021–2022. Six distinct measures of the security of livelihood were developed based on the condition of farmer households in the research location. The index score range for the indicators was 0 to 1. A household that has a higher indication number is likely to be more financially secure and well-off. The security of both health and education are under grave danger. The overall Household Livelihood Security index was found to be 0.54.
Optimizing Mathematics Learning: The Role of Teaching Strategies and Student’s Interests in Performance
Authors: Corda Gemma C.
Published: February 2025

This study assessed the teaching strategies, interests, and performance of first-year college students in Mathematics. Hence, this descriptive-correlational study utilized 381 randomly selected first-year college students. In gathering the data, the study used three valid and reliable researcher-made questionnaires. First, the Mathematical Teaching Strategies Inventory assessed the teaching strategies of teachers as perceived by students. Second, the Mathematical Interest Survey examined the level of interest of students in learning Mathematics. Lastly, the Mathematics Achievement Test determined the students' performance. In analyzing the data, frequency, and percentage were used to profile the respondents. Also, means to assess the teaching strategies, interests, and performance in Mathematics. Meanwhile, Pearson r was used to examine the relationships between these three constructs. Hence, results revealed that the extent of utilization of teaching strategies as perceived by the respondents and the students’ interests in Mathematics when taken as a whole and when they were classified according to the demographics were high. In addition, the performance of first-year college students in Mathematics when taken as a whole and when grouped according to the demographics was satisfactory. Finally, significant relationships existed between the utilization of mathematical teaching strategies and students’ interests; the students’ interests and performance. Meanwhile, no significant relationships were noted between the utilization of mathematical teaching strategies and performance. Given all these findings, various teaching strategies and assessments are encouraged like technology integration and real-life application of the subject that may help invite students' interests and improve the performance of first-year college students in Mathematics.
Cytochrome P450 oxidase activity and its role in NADPH dependent lipid peroxidation
Authors: Bast Aalt , Brenninkmeijer Jan W. , Savenije-Chapel E.Maria , Noordhoek Jan
Published: July 2002

A comparison is made between microsomal NADPH‐dependent H2O2 production and malondialdehyde (MDA) formation in rat liver microsomes, obtained from phenobarbital pretreated rats. An increase in H2O2 formation was observed during NADPH‐dependent disposition (10 min) of 100 μM diazepam (33%) and 2 mM hexobarbital (69%). In contrast orphenadrine (100 μM) and its mono‐N‐demethylated metabolite tofenacine (100 μM) decreased the H2O2 formation (35% and 55%, respectively). However, all these substrates were found to inhibit NADPH‐dependent lipid peroxidation (60 min), estimated by measuring MDA formation, to various extents. These data strongly suggest that the oxidase activity of cytochrome P450 (H2O2 production) is not involved in a rate‐limiting step in NADPH‐dependent lipid peroxidation.
Primary structure of rabbit lung uteroglobin as deduced from the nucleotide sequence of a cDNA
Authors: López de Haro Maria S. , Nieto Antonio
Published: July 2002

Double‐stranded cDNA was synthesized from partially purified uteroglobin mRNA from rabbit lung. A cDNA coding for lung uteroglobin was then cloned in the plasmid pUC18 and both the nucleotide sequence and the derived amino acid sequence were determined. This allowed us to demonstrate unequivocally that uteroglobins from lung and uterus are identical proteins.
Inhibition of carcinogen‐induced cellular transformation of human fibroblasts by drugs that interact with the poly(ADP‐ribose) polymerase system
Authors: Milo George E. , Kurian Ponnama , Kirsten Eva , Kun Ernest
Published: July 2002

Two types of interactions of 13 drugs with human fibroblasts were determined: (a) I
50 of nuclear poly(ADP‐ribose) polymerase, as assayed with isolated nuclei in vitro, and (b) the non‐toxic concentration of drugs that prevented carcinogen‐induced cell transformation of intact fibroblasts (RCF1). In general, RCF1 was much lower than I
50, and one antitransformer did not inhibit the enzyme in vitro, indicating that low‐affinity enzyme inhibitory sites appear to play no role in the mechanism of prevention of cell transformation. Two enzyme inhibitors, caffeine and 1‐methylnicotinamide, exhibited no antitransforming activity. Benzamide when applied in population doubling 1 induced resistance to cell transformation in population doubling 6 by carcinogens added at this stage.
Characterization of the kinetics of neural cell adhesion molecule homophilic binding
Authors: Moran Niamh , Bock Elisabeth
Published: July 2002

A solid‐phase assay has been developed for the investigation of the kinetics of neural cell adhesion molecule (NCAM) binding. Using this assay we can show that NCAM binds to itself in a time‐dependent and saturable manner. Binding constants (K
B values) of 6.9 × 10−8 M and 1.23 × 10−6 M, respectively, were obtained for adult and newborn rat NCAM homophilic binding. Binding is specifically inhibited by Fab′ fragments of polyclonal anti‐NCAM antibodies but is unaffected by heparin or chondroitin sulphate. This indicates that the NCAM homophilic binding site is separate from and independent of the heparin‐binding site and that a developmental modification, probably polysialation, gives rise to marked differences in the adhesive properties of NCAM.
Vitamin K1 (phylloquinone) restores the turnover of FeS centers in the ether‐extracted spinach PS I particles
Authors: Itoh Shigeru , Iwaki Masayo
Published: July 2002

The effects of vitamin K1 (phylloquinone) addition on the turnover of the FeS centers in photosysten, I photochemistry were studied in diethyl ether‐extracted spinach photosystem I particles. Reconstitution of one Molecule of vitamin K1/reaction center was sufficient to suppress the charge recombination between the oxidized reaction center chlorophyll P700+ and the reduced electron acceptor intermediate chlorophyll a and to restore the turnover of FeS centers in the ether‐extracted particles. This strongly suggests that reconstituted vitamin K1, functions the primary electron acceptor A1 and exhibits a redox midpoint potential low enough to reduce the FeS centers. The quinone binding site in photosystem I, which enables vitamin K1 to show such a low redox potential, seems to be more hydrophobic than those in reaction centers of photosystem II or purple bacteria.
The hexose carrier from Chlorella
Authors: Sauer Norbert , Tanner Widmar
Published: July 2002

The cDNA coding for the inducible H+/hexose cotransporter of Chlorella kessleri has been cloned and sequenced. It was isolated by differential screening of a cDNA library prepared from glucose‐induced cells. The increase in expression of the gene correlates quantitatively with the increase in uptake activity due to induction; it is not expressed in a hexose transport mutant. An open reading frame allows for a membrane protein of 533 amino acids with a relative molecular mass of 57 kDa. This protein is highly homologous to the human and rat glucose transporters catalyzing facilitated diffusion and to the bacterial H+/pentose cotransporters. It is not related to the H+/lactose cotransporter of E.colli and to the mammalian Na+/glucose cotransporter.
Sequence location of a putative transglutaminase cross‐linking site in human vitronectin
Authors: Skorstengaard Karna , Halkier Torben , Højrup Peter , Mosher Deane
Published: July 2002

We attempted to locate the glutamine residue in human vitronectin, susceptible to cross‐linking by transglutaminases. Vitronectin was incubated with 14C‐labelled putrescine and plasma factor XIIIa and, after reduction and alkylation, the vitronectin was digested with trypsin. HPLC of the digest followed by scintillation counting revealed one major and two minor radioactivity labelled peaks. Sub‐digestion with Staphylococcus aureus protease, sequence analysis and mass‐spectrometry of the resulting peptides demonstrated that Gln‐93 of vitronectin had incorporated putrescine. Additionally, Gln‐73, Gln‐84 and Gln‐86 were found to be minor sites for incorporation.
Determination of DNA binding specificities of mutated zinc finger domains
Authors: Thiesen Hans-Jürgen , Bach Christian
Published: July 2002

By substituting non conserved amino acids present in the postulated α‐helical region of zinc finger domains, we demonstrated that Cys2/His2 type zinc finger domains could be targeted to new DNA binding sites. The putative α‐helical region of the second SPI zinc finger (SLH) was replaced by amino acids (SLH) occurring in analogous zinc finger positions of human zinc finger protein Kox 29. The DNA binding specificity of the FPLC purified chimaeric protein (SPI‐Kox 29) was determined by use of the target detection assay (TDA). Chimaeric protein SPI‐Kox 29 wax subjected to randomized oligonucleotides (GGG NNNN GGC) that were designed on the basis that each SPI zinc finger interacts with 3–4 nucleotides concerning its cognate target site GGG GCGG GGC. By this analysis the DNA binding specificity of SPI‐Kox 29 was shown to have switched from the cognate SPI binding site to GGG GGTG GGC. Structure‐function analysis of this type should facilitate the determination of DNA binding specificities for any individual zinc finger of interest.
Identification of a 220 kDa membrane‐associated plant cell protein immunologically related to human β‐spectrin
Authors: Michaud D. , Guillet G. , Rogers P.A. , Charest P.M.
Published: July 2002

Electrophoretic analysis of low ionic strength extracts of tomato plant leaves revealed the presence of two proteins of apparent molecular weights of 240 kDa and 220 kDa which co‐migrated with purified human erythrocyte α‐ and β‐spectrin subunits. Immunochemical analyses employing an affinity‐purified polyclonal antibody to human erythrocyte β‐spectrin reacted specifically with the 220 kDa plant cell protein. Immunofluorescence microscopy indicated that the β‐spectrin antibody recognized an antigen which was primarily restricted to the peripheral areas of the cells. Collectively, these results suggest that the cells of higher plants contain polypeptides related to the spectrin family of proteins. It is proposed that the plant cell possesses a membrane skeleton which is structurally and perhaps functionally analogous to that of the animal cell.
Phosphoprotein phosphatase 2A dephosphorylates eIF‐4E and does not alter binding to the mRNA cap
Authors: Bu Xin , Hagedorn Curt H.
Published: July 2002

The phosphorylation and dephosphorylation of the 25 kDa mRNA cap binding protein eukaryotic initiation factor‐4E (eIF‐4E) is regulated during different physiologic and pathophysiologic states that include cell growth and the late phase of adenovirus infection. We have found that okadaic acid is much more effective in increasing the phosphorylated fraction of eIF‐4E than phorbol 12‐myristate 13‐acetate in Hep G2 cells. Phosphoprotein phosphatase 2A dephosphorylated eIF‐4E isolated from both phorbol 12‐myristate 13‐acetate‐ or okadaic acid‐treated cells, whereas alkaline and acid phosphatase were relatively ineffective. The ability of purified [35S]eIF‐4E isolated from okadaic acid‐treated cells to bind mRNA caps was compared to phosphoprotein phosphatase 2A‐treated [35S]eIF‐4E and found to be no different. This suggests that alternative explanations for the previously observed effects of eIF‐4E phosphorylation on protein synthesis must be considered. In addition, our results indicate that the in vivo phosphorylation eIF‐4E is not catalyzed solely by protein kinase C.
Isolation of factor Xa from chick embryo as the amniotic endoprotease responsible for paramyxovirus activation
Authors: Gotoh Bin , Yamauchi Fumiko , Ogasawara Tomohiko , Nagai Yoshiyuki
Published: July 2002

In chick embryo, certain paramyxoviruses mainly target the chorioallantois and the allantoamnion and show no extensive further spreading in the other organs. This has been explained by the possible presence of an endoprotease activating the viral fusion glycoprotein precursor in the allantoic and the amniotic fluid, and its absence in other places or organs, We previously isolated such an endoprotease from the allantoic fluid and demonstrated its identity with the clotting factor Xa. Exactly the same endoprotease by all the criteria including the N‐terminal amino acid sequence was now isolated from the amniotic fluid. Thus, the factor Xa seems to be a major host determinant of the viral tropism in chick embryo.
Pro‐oxidants and mitochondrial Ca2+: their relationship to apoptosis and oncogenesis
Authors: Richter Christoph
Published: July 2002

Apoptosis is a physiological process for active cell removal. One of its hallmarks is an increased cytosolic Ca2+ content. Several genes involved in apoptosis control have been identified, but their mode of action is not understood in detail. Apoptosis may relate to oncogenesis, in that some malignant tumors may grow because genes engaged in apoptosis control are altered. L929 cells overexpressing the proto‐oncogen bcl‐2 have an increased mitochondrial membrane potential (δψ), as have many carcinoma cells, bcl‐2 protects L929 cells against apoptosis caused by pro‐oxidant‐induced mitochondrial Ca2+ ‘cycling’ and increased cytosolic Ca2+ levels. Nerve growth factor, which induces catalase, and inhibitors of mitochondrial Ca2+ release also prevent apoptosis. It is suggested that a pro‐oxidant‐induced Ca2+ release from mitochondria, followed by Ca2+ cycling and ATP depletion, is a common basic event during apoptosis. Accordingly, maintenance of δψ stabilizes mitochondria, thereby prevents apoptosis, and may confer increased growth potential to cells.
Calcium‐sensitive control of rhodopsin phosphorylation in the reconstituted system consisting of photoreceptor membranes, rhodopsin kinase and recoverin
Authors: Gorodovikova Elena N. , Senin Ivan I. , Philippov Pavel P.
Published: July 2002

Rhodopsin phosphorylation in the reconstituted system consisting of urea‐washed photoreceptor membranes, rhodopsin kinase and recoverin is regulated by Ca2+: the process takes place at low [Ca2+] but is suppressed at high [Ca2+]. In the absence of recoverin, rhodopsin kinase is active irrespective of the cation concentration used. Hence, recoverin is an inhibitor (at high [Ca2+]) but not an activator of rhodopsin kinase. Based jointly on these data obtained on the reconstituted system and on our preceding experiments on rod outer segments suspension, one may conclude that (i) the function of recoverin in retina rod cells is the Ca2+‐sensitive control of rhodopsin phosphorylation and (ii) the presence of recoverin is essential and sufficient to provide rhodopsin kinase with the Ca2+ sensitivity.
Current and future strategies for insomnia management
Authors: López-Ibor JJ
Published: July 2002

Different forms of insomnia are present as symptoms of many psychiatric and other disorders. The first step for a treatment strategy is therefore a correct diagnosis. As insomnia is more common in patients with psychiatric disorders than in the general population, a careful consideration should be given, depressive and anxiety disorders should especially be carefully investigated. There are reasons to believe that even in so-called insomnia not obviously related to psychiatric disorders, stressful life situations may play a role. Therefore a co-morbidity with emotional disorders which may follow these events is worth considering. Insomnia should always be considered as pan of a sleep-wake schedule disturbance and this has an impact on the importance of the pharmacological properties of the drugs used to treat insomnia. The recent trend for more specific agents both on receptor sub-populations and on relevant sites of the GABA receptor complex will help very much in selecting the most appropriate drug for treating patients.
Measuring patient benefit in mental illness
Authors: Gudex C
Published: July 2002

This paper provides a brief overview of issues to consider in evaluating patient benefit from health care. Different types of measures are described and it is noted that the choice of measure is dependent upon the purpose of the study. The cost-utility approach to evaluation is discussed in more detail. This approach is often criticised but it raises important issues in medical decision-making and has considerable potential in the evaluation of patient benefit from health care.
HSF1/hsp72 pathway as an endogenous anti‐inflammatory system
Authors: Ianaro Angela , Ialenti Armando , Maffia Pasquale , Pisano Barbara , Di Rosa Massimo
Published: July 2002

We investigated the occurrence and the role of HSF1 activation and inducible hsp72 expression in the carrageenin pleurisy in the rat. Molecular analysis performed on pleural cells collected from rat pleural cavity after carrageenin challenge revealed increased HSF1 activation and hsp72 expression. Moreover, local injection of a double‐stranded oligodeoxynucleotide (ODN) containing the heat shock element sequence, acting as transcription factor decoy, exacerbated the inflammatory reaction. The exacerbation, induced by wild‐type, but not by mutant ODN decoy, was associated to both inhibition of HSF1/DNA binding activity and reduction of hsp72 expression. In conclusion, this study shows that HSF1 activation and hsp72 expression both actually occur in acute inflammation and that the remission of the inflammatory reaction is tightly associated to the HSF1‐dependent hsp72 expression, suggesting a a relevant role for the HSF1/hsp72 pathway as an endogenous anti‐inflammatory system.
A study on reducing substrates of manganese‐oxidizing peroxidases from Pleurotus eryngii and Bjerkandera adusta
Authors: Heinfling Annette , Ruiz-Dueñas Francisco Javier , Martı́nez Marı́a J , Bergbauer Matthias , Szewzyk Ulrich , Martı́nez Angel T
Published: July 2002

A novel peroxidase, oxidizing Mn2+ and different aromatic compounds, was isolated. Hydroquinones, substituted phenols, dyes, other aromatic compounds and Mn2+ were compared as reducing substrates, and conclusions presented in the light of a molecular model built by homology modeling. The enzymes showed the fastest reaction rates with Mn2+, but the highest affinity corresponded to hydroquinones and dyes. Oxidation of Reactive Black 5 (an azo‐dye not oxidized by Mn3+) was non‐competitively inhibited by Mn2+. These findings, together with identification of putative Mn‐binding site (involving Glu36, Glu40, Asp175 and inner heme propionate) and long‐range electron transfer pathways, indicate that different sites are involved in substrate oxidation.
Unexpected expression of carbonic anhydrase I and selenium‐binding protein as the only major non‐heme proteins in erythrocytes of the subterranean mole rat (Spalax ehrenbergi)
Authors: Yang Hong , Nevo Eviatar , Tashian Richard E
Published: July 2002

Chromatographic separation of the non‐heme proteins from the erythrocytes of the subterranean mole rat belonging to the superspeciesSpalax ehrenbergifrom Israel revealed two major peaks. On sequence analyses, the larger peak corresponded to a 56 kDa selenium‐binding protein (SeBP) previously characterized from mouse and human liver, and the second peak to the low‐activity carbonic anhydrase (CA) isozyme, CA I. There was no evidence of the high‐activity CA II isozyme normally found in the red cells of all amniotes tested to date. Thus, the mole rat appears to be the first mammalian species to express both a SeBP and the low‐activity CA I isozyme, as the major non‐heme proteins in its red blood cells. It is possible that the absence of the high‐activity CA II isozyme may be advantageous to the mole rat in adapting to the low O2and high CO2environment of its underground burrows. It is also likely that the 56 kDa SeBP may play an important adaptive role in the physiology of the red cell.
Identification of a cytochrome P450 cDNA encoding (2S)‐flavanone 2‐hydroxylase of licorice (Glycyrrhiza echinata L.; Fabaceae) which represents licodione synthase and flavone synthase II1
Authors: Akashi Tomoyoshi , Aoki Toshio , Ayabe Shin-ichi
Published: July 2002

The microsome of insect cells expressing CYP Ge‐5 (CYP93B1), a cytochrome P450 cDNA of licorice (Glycyrrhiza echinata L.), catalyzed the formation of [14C]licodione and [14C]‐2‐hydroxynaringenin from (2S)‐[14C]liquiritigenin and (2S)‐[14C]naringenin, respectively. On acid treatment, the products were converted to 14C‐labeled 7,4′‐dihydroxyflavone and apigenin. Eriodictyol was also converted to luteolin by the reaction with the microsome of yeast expressing CYP93B1 and subsequent acid treatment. CYP93B1 was thus shown to encode (2S)‐flavanone 2‐hydroxylase, which has previously been designated to licodione synthase and flavone synthase II depending on the substrates employed.
Purification, molecular properties and specificity of a thermoactive and thermostable proteinase from Pyrococcus abyssi, strain st 549, hyperthermophilic archaea from deep‐sea hydrothermal ecosystem
Authors: Dib Robert , Chobert Jean-Marc , Dalgalarrondo Michèle , Barbier Georges , Haertlé Thomas
Published: July 2002

A protease was isolated and purified from the supernatant of a culture of hyperthermophilic archaebacteria: Pyrococcus abyssi strain st 549. Purification consisted of three chromatographic steps. The enzyme purification yield was 4% and the purification factor 890. This protease is a seryl‐protease hydrolyzing proteins and peptides with a preference for cleavage at the aromatic and hydrophobic residues in P1 and P′1 positions. Its activity is optimal at 95°C and at pH 9. The electrophoretic mobility of the protease observed by zymogram suggests that it can adopt several oligomer forms. Three of them predominate displaying apparent molecular masses of 150, 105 and 60 kDa. Interdependence of the observed bands was revealed by changing the denaturation conditions of the samples (temperature, SDS concentration) before electrophoresis.