bluuebunny/crossref_metadata_2025 huggingface dataset:
Domesticating and Commercialisation of Indigenous Fruit and Nut Tree Crops for Food Security and Income Generation in the Kingdom of Eswatini
Authors: Kunene Emmanuel N. , Nxumalo Kwanele A. , Ngwenya Menzi P. , Masarirambi Michael T.
Published: June 2020

Indigenous fruit and nut tree crops are very important to the human diet and subsequent food security and income generation in the Kingdom of Eswatini. Indigenous fruits and nuts are important for the nutritional contribution as sources of Vitamins and minerals. They also provide fibre which is essential for proper digestion of food. Fruits, particularly nuts provide man with unsaturated fats which are good for the heat and the cardio vascular system. Indigenous fruits and nuts are aesthetic as they provide interesting colour, texture and variety. Despite the immense benefits of indigenous fruits and nuts to the people of the Kingdom of Eswatini, there has not been enough effort put towards domesticating the indigenous fruit and nut trees and subsequent commercialising their products. The objective of this paper was to document strategies towards domestication and commercialisation of indigenous fruits and nuts for improved livelihoods of Emaswati citizens, which in turn will help achieve sustainable development goals (SDGs) pertaining to human health and food security in the Kingdom of Eswatini. It can be concluded that domesticating indigenous fruit and nut trees can be achieved through a multi-stakeholder involvement. Domestication can also help in conservation of indigenous fruit and nut trees. Value addition on indigenous fruits and nuts can generate more income.
Soil Carbon Pools, Carbon and Nitrogen Storage Pattern in Soil Aggregate Fractions under Long-term Application of Organic and Synthetic Fertilizers in Rice-Wheat System: A Review
Authors: Yadav Shipra , Naresh R. K. , Vivek . , Chandra M. Sharath , Mahajan N. C.
Published: June 2020

Soil organic matter (SOM) has long been recognized as an important indicator of soil productivity. The SOM refers to the organic fraction of undecayed plant and animal residues. The preservation of SOM is crucial to ensure long-term sustainability of agricultural ecosystems. OM plays a critical role in the global carbon balance that is thought to be the major factor affecting global warming. Overall, adequate amounts of SOM maintain soil quality and reduce environmental pollution. SOC concentrations and storage to 60 cm depth are significantly influenced due to long-term fertilization. The SOC storage in 0–60 cm in NP+FYM (farmyard manure), NP+S, FYM and NP treatments were increased as compared to the CK treatment. The concentration of particulate organic carbon (POC), dissolved organic carbon (DOC) and microbial biomass carbon (MBC) in organic manure plus inorganic fertilizer treatments (NP+S and NP+FYM) in 0–60 cm depth increased linearly with increasing SOC content. Light fraction organic carbon (LFOC) was also significantly higher following the treatments including organic amendment than following applications solely of chemical fertilizers. Application solely of chemical fertilizers had no significant effects on LFOC and KMnO4C fractions compared with unfertilized control plots. Carbon pools were significantly correlated with SOC, which increased with application of organic amendments. Threshold C input of 3.3 MgCha-1 yr-1 was needed to maintain the SOC stock even at the low antecedent level. This review study will be helpful in crafting sustainable nutrient management programs in the future to enhance crop productivity with high efficiency and minimum nutrient loss. Therefore, fertilization strategies that include organic manure can increase the pool of stable C in the surface soil layer, while at the same time increasing concentrations and proportions of labile C. Organic manure use contributes to improved nutrient cycling services and higher soil quality in rice-wheat system.
Pharmacognostic Study of Justicia beddomei (C. B. Clarke) Bennet
Authors: Haleshi C. , Sringeswara A. N. , Danapur Vijay
Published: June 2020

Justicia beddomei (C. B. Clarke) Bennet synonym Adhatoda beddomei C.B. Clarke consists of the fresh or dried leaves Dried leaves are of a dull brownish-green colour, characteristic odour and bitter taste. The leaves of the plant contain major alkaloids called vasicine, vasicinone and deoxyvasicinone, sitosterol and its glucoside, which are shown to be having bronchodilator and antihistaminic effects. These alkaloids are said to exist in combination with an acid that has been named adhatodic acid. Leaves yield essential oil and an alkaloid vasicine. Roots contain vasicinol and vasicinone. Roots also contain vasicoline, adhatodine, anisotine and vasicolinone. Several alkaloids like quinazoline and valicine are present in this plant. The leaf extracts of Justicia beddomei (C. B. Clarke) Bennet is used as an expectorant especially in chronic bronchitis and asthma. It relieves cough and breathlessness. It is also prescribed commonly for local bleeding due to peptic ulcer, piles etc. Its local use gives relief in pyorrhoea and in bleeding gums. In the present study, detailed Pharmacognosy of this plant is studied as it is used as substitute for Adathoda vasica.
The Effect of Market Performance, Corporate Governance and Examination of Corporate Taxes with Tax Amnesty as Moderators
Authors: Faisal Aris Riantori , Murwaningsari Etty , Mayangsari Sekar , Mulyani Susi Dwi
Published: June 2020

The purpose of this study is to examine and analyze the effect of market performance, corporate governance, tax audits toward corporate tax, and the role of the tax amnesty in moderating the relationship between market performance, corporate governance, and tax audits toward corporate tax. The unit of analysis is a public company (corporate taxpayers), with a purposive sampling sample of manufacturing industry companies listed on the Indonesia Stock Exchange and meeting the sampling criteria of 52 companies in the 2014-2017 period.The results of the study are that market performance is not proven to have a positive effect on corporate tax, this shows that the higher market performance as measured by Tobin’s Q does not have an impact on corporate tax increases; Corporate governance is not proven to have a positive effect on corporate tax, this shows that the higher corporate governance does not automatically raise corporate taxes; Tax audits have a positive impact on corporate taxes, this indicates that the higher the examination of taxes carried out, the corporation tax will increase; Tax amnesty has not been proven to strengthen the positive influence of market performance, corporate governance and tax audits of corporate taxes. For the sensitivity model with ETR, none of the proposed hypotheses has been proven.
Export Performance and Trade Direction of Fresh and Dried Grapes (Raisin): Evidence from Afghanistan
Authors: Mushair Hasibullah , Murugananthi D. , Rohini A.
Published: June 2020

Grapes are one of the most delicious refreshing and nourishing fruits. Afghanistan produces 9.84 lakhtonnes of Grapes during 2018 (FAO statistics). It is one of the important commodities in the export basket of Afghanistan. The major export markets for Afghan’s fresh grapes are Pakistan and India. The major export markets for dried grapes (raisin) are India, Russia, Belarus, USA, Pakistan and UK. The present study aims to quantify the export performance and changing the structure of Grapes and raisin exports from Afghanistan. Secondary data on area, production and country-wise quantity exports of fresh and dried grapes were collected from FAO statistics, ITC and APEDA for a period of 13 years from 2006 to 2018. Compound Annual Growth Rate was computed for studying the trend in the area, production, yield, export quantity and export value for fresh and dried (raisin) grapes. Markov chain analysis was attempted to assess the direction of change in exports. Markov chain analysis results showed that Pakistan is the stable market for Afghan fresh Grapes and India and France are less stable markets. The major reasons are a geographical advantage for Pakistan which gave a competitive advantage over other countries concerning fresh grapes. India, Russia and Pakistan are stable markets for dried grapes and USA, UK, Germany and Belarus are less stable markets. India is the main country to import dried grapes (raisin) in the next five years. It shows a high value in terms of quantity and percentage which is more than 50 percent of all Afghanistan's dried grapes (raisin) export.
Multilingualism as Curriculum Policy in Cameroon Education System
Authors: Loveline Yaro
Published: June 2020

The concern on the effective inclusion of indigenous languages in the school curriculum has been central to educational discussions among scholars and stakeholders. It is true that language as a medium of communication is very vital for sustainable growth in any society. However, the level of implementation of the policy of multilingualism in classrooms in Cameroon poses so many pedagogical challenges. While the Government is collaborating with private agencies such as PROPELCA(Cameroon National Language Association) in introducing the teaching of national languages in primary schools,  and though the Government through the Ministry of Basic Education has also created an inspectorate in charge of the introduction of indigenous languages in Primary schools, there still exist some resistance from stakeholders to introduce the teaching of native languages in the curriculum of schools despite recommendations made in the 1995 education forum and the provision in article 4 section 5 of the 1998 Education Law. The study argues that, since the introduction of multilingualism as curriculum policy in Cameroon, its implementation is still at the pilot stage. Using the Historical analytic approach, data was collected from primary and secondary sources in establishing the level of use of the curriculum policy of multilingualism as a guide to teaching and learning in schools in Cameroon.
Variation Analysis of Material Characteristics of Steel through Mechanical Calibration
Authors: Kim Sung Bae , Kim Sang Seup , Baek Ki Youl
Published: June 2020

Steel elements exhibit varying characteristics during site installation after shop manufacturing. There may be a mix-up owing to unclear management methods for transport, and deformation might occur during field installation. Moreover, the technical basis for evaluating repair methods is narrow. In this study, variations in material properties after performing mechanical calibration were determined for widely used steel types, such as SS400 and SM490, in which thicknesses of 30 mm and less are frequently used on sites. The test results showed that the mechanical calibration in unheated and thermal heating cases increased the yield strength and tensile strength of steel, and the Korea Standard value was satisfied. A yield ratio of approximately 85% or less was obtained, indicating sufficient toughness. In addition, variations owing to thermal heating did not occur when temperatures below 650 °C were applied.
A Suggested Method for Performance Evaluation of Large Scale Fireproof Shutters
Authors: Woo Youngje
Published: June 2020

Currently, there is a limitation in the performance evaluation of a large fireproof shutter due to the constraint of the maximum size of the test equipment (3 m × 3 m). In particular, the longer the maximum span of a large fireproof shutter, the greater the fire load. Accordingly, the deformation of the large fireproof shutter is increased. The fire fragile area between the guide rails at both ends of the large fireproof shutter and the body of the fireproof shutter increases, and at the same time, the possibility of flame passing through the fragile area increases. However, the reduced-scale test is performed without reflecting the above-mentioned problems, and the maximum span of 8 m specified in KS F 4510 is widely used. Therefore, the author suggests a new method for performance evaluation of large-scale fireproof shutter, which can reflect the fire vulnerabilities of large fireproof shutter with increasing span.
Performance-Based Evaluation on Egress Safety of Nursing Hospitals Considering Egress Guides and Delay Time
Authors: Lee Doohee , Heo Inwook , Khaliunaa Darkhanbat , Han Sun-Jin , Kim Kang Su
Published: June 2020

In recent years, the number of nursing hospitals has significantly increased as the number of elderly has increased. However, since there are many patients who have difficulty in moving quickly in the nursing hospitals, it is very hard to evacuate rapidly in the event of a fire, which results in many casualties. In this study, fire and evacuation simulations for the nursing hospital were performed, based on which the evacuation safety was evaluated in detail according to the location of the fire compartment, number of evacuation guides, and evacuation delay time. The simulation results showed that it is necessary to secure the number of evacuation guides complying with the legal standards, and to reduce the evacuation delay time to be less than 60 s in order to avoid loss of life.
A Study on the Efficiency of Vertical Non-Combustible Blocking for External Fire Spread of EIFS with Combustible Insulation in Piloti Building
Authors: Cho Kyungsuk , Chae Seungun , Choi Jihun , Cho Bumyean
Published: June 2020

The requirements for exterior finishing materials have been substantially established recently. After a fire at an apartment building in Uijeongbu City, fire safety requirements for exterior finishing materials used in the buildings that have six and more stories or are taller than 22 meters were specified in 2015, and three and more stories or are taller than 9 meters were specified in 2019. However, the buildings built before that may not accomplish the safety requirements and thus have fire safety problem. This study suggests a method for partially reinforcing the Exterior Insulation Finishing System (EIFS) of buildings to enhance the fire safety. EIFS specimens whose walls on the pilotis were reinforced with a mineral insulation material, were tested in accordance with BS 8414-1. The result showed that fire spread was delayed more than 9 min when compared with an EIFS specimen without reinforcement.
The Improvement of Safety Inspection Policies for Passenger Ships, Excursion Ships, and Ferries through Safety Management Analysis and On-site Inspections
Authors: Park Hayong , Woo Juhyoung , Yi Waonho
Published: June 2020

In contrast to land, sea routes are exposed to many dangers due to certain conditions on the surface of the water. In the wake of the Sewol ferry disaster in April, 2014, ferry and other ship-related systems were improved. Experts from related organizations have since conducted safety inspections on passenger ships, excursion ships, and ferries in accordance with the National Security Agency and individual laws. These inspections were introduced after the public's attention was raised from not only the Sewol incident but also from the sinking of the cruise ship Hableány, which happened in Budapest, Hungary on May 29, 2015. Accordingly, this study presents measures to improve the safety management of passenger ships, excursion ships, and ferries such as facility standards, inspection regulations, and safety management regulations. These tasks will be accomplished by comprehensively analyzing problems through on-site inspections combined with safety management initiatives which will identify safety issues regarding passenger ships, excursion ships, and ferries.
Experimental Study on the Temperature Distribution and Residual Strength of Fire-damaged RC Columns Depending on Heated Areas
Authors: Ryu Eunmi , Kim Heesun , Shin Yeongsoo
Published: June 2020

Most of the columns in actual fire conditions are heated on partial faces rather than all four sides due to the floor plan, which results in asymmetric behaviors of columns. The asymmetric behaviors of fire-damaged columns may cause more vulnerability to the structural performance. In this study, temperature distribution and residual strength of reinforced concrete columns exposed to fire were investigated according to various heated areas. To achieve the objective, columns were heated for 2 h according to ISO-834 standard time-temperature curve and subsequently tested under the axial loading after a week. The test results show that the residual strength of the fire-damaged columns decreased as the heated area increased, and the residual strength reduced additionally due to asymmetric heating.
An Experimental Study of Vertical Fire Spread through Openings in Compartments
Authors: Kweon Ohsang , Kang Hyun
Published: June 2020

In this study, a real scale fire experiment was conducted to analyze the characteristics of the flame that is ejected from an opening. The experiment was conducted by setting mattresses and wood cribs in compartments sized 2.4 (L) × 3.6 (W) × 2.4 (H) m and using heptane as an ignition source, and changes in external heat flux and temperature were measured during the experiment. In the experiment, openings of two sizes: small [2.2 (L) × 1.0 × 1.0 (H) m] and large [2.2 (L) × 2.0 × 2.0 (H) m], were used. The results showed that the maximum decrease of 40.7 kW/㎡ and 31.7 kW/㎡ was measured in small and large openings, respectively, at the distance of 0.5 m from the top of the opening. As a test result, securing the separation distance of the upper and lower openings can greatly affect the prevention of vertical spread of the flame, and if sufficient separation distance is not secured, it is considered that an additional fire diffusion prevention method is necessary.
A fucosyltransferase in teratocarcinoma stem cells
Authors: Muramatsu Hisako , Muramatsu Takashi
Published: July 2002

Teratocarcinoma stem cell F9 expressed a potent fucosyltransferase activity acting on asialofetuin. A majority of the product was susceptible to α‐L‐fucosidase I from almond emulsin, indicating that the linkage formed was mainly Fucα1 → 3GlcNAc. The specific activity of the transferase decreased when the stem cells were induced to differentiate into parietal endoderm cells by retinoic acid and dibutyryl cyclic AMP. Furthermore, PYS‐2 cell, a parietal endoderm cell line virtually lacked the transferase. The change in the fucosyltransferase activity could be correlated with cell surface changes occurring during differentiation.
Rat glutathione transferase 8‐8, an enzyme efficiently detoxifying 4‐hydroxyalk‐2‐enals
Authors: Jensson Helgi , Guthenberg Claes , Ålin Per , Mannervik Bengt
Published: July 2002

Rat glutathione transferase 8‐8 is one of the less abundant cytosolic glutathione transferases, accounting for approx. 1% of the total activity with 1‐chloro‐2,4‐dinitrobenzene in liver. The enzyme is eluted at pH 6.3 upon chromatofocusing and has so far been identified in liver, kidney, lung and testis. Characteristic properties include high relative activity with ethacrynic acid (70% of the specific activity with 1‐chloro‐2,4‐dinitrobenzene) and an apparent subunit M
r of 24500. The most significant property noted is the high catalytic activity in the conjugation of 4‐hydroxyalk‐2‐enals, major products of lipid peroxidation. The catalytic efficiency with these substrates exceeds corresponding values for all known substrates tested with any glutathione transferase, which suggests that transferase 8‐8 may have evolved to detoxify 4‐hydroxyalk‐2‐enals.
A mechanism for the RNA‐catalyzed formation of 5′‐phosphates The origin of nucleases
Authors: Cedergren Robert , Lang B.Franz , Gravel Denis
Published: July 2002

Processes involved in RNA metabolism can be distinguished by the nature of the sugar phosphate substitution (5′ or 3′) in intermediates or products. Although it is known that 3′‐phosphates are produced via a 2′,3′‐cyclic phosphate intermediate, formed by nucleophilic attack on the phosphodiester bond by the adjacent 2′‐OH, little is known about the production of 5′‐phosphate products. We attribute 5′‐phosphate intermediates and products to a preferred configuration of the pentavalent phosphorus intermediate resulting from the attack of a distant nucleophile. This intermediate is favored, since its formation is possible without major conformational changes in the molecule. Based on the two products of nucleic acid hydrolysis we define: the conjunct and disjunct nucleophile mechanisms, each of which would have independent origins. Indeed, the products of an overwhelming number of nucleases and RNases are consistent with one of these mechanistic models demontrating that the origin of these enzymes are deeply rooted in the intrinsic chemistry of phosphate esters.
The conformation of calmodulin: A substantial environmentally sensitive helical transition in Ca4‐calmodulin with potential mechanistic function
Authors: Bayley Peter , Martin Stephen , Jones Gareth
Published: July 2002

The conformation of Ca4‐calmodulin in solution, as assessed by far‐UV peptide circular dichroism, contains significantly less α‐helix than the proposed X‐ray crystal structure. We now show that Ca4‐calmodulin adopts significant additional helical structure in solution in the presence of a helicogenic solvent (50%, v/v, aqueous 2,2,2‐trifluoroethanol or 50%, v/v, methylpentane‐5,5‐diol). We suggest that the long continuous helix (residues 66–92 of the crystal structure)is not necessarily a normal feature of the calmodulin structure in solution, and may be due in part to the conditions of crystallisation. This result is supported by time‐resolved tyrosine fluorescence anisotropy studies indicating that Ca4‐calmodulin in solution is an essentially compact globular structure which undergoes isotropic rotational motion. We conclude that, under appropriate ionic and apolar environmental conditions, Ca4‐calmodulin undergoes a substantial helical transition, which may involve residues in the central region of the molecule. Such a transition could have an important function in determining specificity and affinity in interactions of calmodulin with different target sequences of Ca2+‐dependent regulatory enzymes.
Heparan sulphate with no affinity for antithrombin III and the control of haemostasis
Authors: Scully Michael F. , Ellis Vincent , Kakkar Vijay V.
Published: July 2002

Heparan sulphate with no affinity for antithrombin III (ATIII) was observed to cause acceleration of the factor Xa:ATIII interaction by 1100‐fold (k
2, 7 × 107 M−1·min−1) and the prothrombinase:ATIII interaction by 2900‐fold (k
2, 2.5 × 107 M−1·min−1). Although high‐affinity heparan sulphate catalyzed higher acceleration and at lower concentration, in natural mixtures of the two forms the activity of the no affinity form predominated. Heparan sulphate had no significant effect on the thrombin:ATIII interaction but inhibited its potentiation by heparin (K
d 0.3 μM). From the estimated concentration of heparan sulphate on the endothelial cell surface it is proposed that the non‐thrombogenic property of blood vessels is due to the acceleration of the factor Xa or prothrombinase:ATIII interaction by the greater mass of surface‐bound heparan sulphate rather than by the much smaller proportion of heparin‐like molecules (with high affinity for antithrombin III) which may be present.
The angiotensinogen gene is located on mouse chromosome 8
Authors: Clouston W.M. , Fournier R.E.K. , Richards R.I.
Published: July 2002

We have recently identified a cis‐acting genetic lesion affecting angiotensinogen gene expression in testis and salivary gland. Accordingly, the angiotensinogen gene was assigned to mouse chromosome 8 by screening a series of hybrid cell lines for retention of mouse angiotensinogen sequences by genomic Southern analysis. In AKXD recombinant inbred mice, the angiotensinogen gene is 2.4 ± 1.8 centiMorgan from Rn7S‐8, a 7S RNA gene located on chromosome 8 (Taylor, B.A., personal communication). However, the segregation of salivary and testicular angiotensinogen expression phenotypes into inbred mouse strains was not concordant with the known chromosome 8 proviruses Emv‐2, Mtv‐21, Xmv‐12 or Xmv‐26.
The structural gene for cytochrome c551 from Pseudomonas aeruginosa
Authors: Nordling Margareta , Young Simon , Karlsson B.Göran , Lundberg Lennart G.
Published: July 2002

The gene coding for Pseudomonas aeruginosa cytochrome c
551 has been cloned and its nucleotide sequence determined. Cytochrome c
551 is expressed as a 104 amino acid pre‐proteinfrom which a signal peptide of 22 amino acids is cleaved off during the translocation across the cytoplasmic membrane. The gene is located just downstream of the gene coding for nitrite reductase on the Pseudomonas aeruginosa chromosome, suggesting that these genes form an operon.
Specific interaction of the new fluorescent dye 10‐N‐nonyl acridine orange with inner mitochondrial membrane
Authors: Maftah Abderrahman , Petit Jean Michel , Julien Raymond
Published: July 2002

The fluorescent dye 10‐N‐nonyl acridine orange (NAO), known as specifically associated with mitochondria, has been reported to have a cytotoxic effect when high doses were applied to cells. Presently, the biochemical basis of its toxicity was investigated on isolated rat liver mitochondria. At low concentrations, NAO strongly inhibited state 3 respiration and ATP synthesis. At high concentrations, electron transport, ATP hydrolysis, Pi‐transport and adenine nucleotide activities were also decreased. All these inhibitions can be explained by probe‐cardiolipin interactions which could induce the collapse of energy conversion and/or the modification of membrane fluidity.
Surface membrane CD4 turnover in phorbol ester stimulated T‐lymphocytes
Authors: Møller B.K. , Andresen B.S. , Christensen E.Ilsø , Petersen C.Munck
Published: July 2002

Down‐regulation of surface membrane CD4 (smCD4) in phorbol ester stimulated T‐cells resulted from internalization. Internalization (T

= 15 min at 50 ng ) was followed by degradation of CD4‐bound antibodies. Degradation in unstimulated T‐cells was comparatively insignificant. Release of degradation products was PMA dose‐dependent and could be inhibited by methylamine. Uptake and degradation continued after maximal down‐regulation of surface membrane CD4, and methylamine did not inhibit reappearance of smCD4 antigens. Metabolic labelling of T‐cells further showed that ongoing synthesis rather than recycling contributed to an accelerated smCD4 turnover in activated cells.
The antioxidant action of tamoxifen and its metabolites Inhibition of lipid peroxidation
Authors: Wiseman Helen , Laughton Miranda J. , Arnstein Henry R.V. , Cannon Michael , Halliwell Barry
Published: July 2002

The anti‐oestrogen drug tamoxifen is an inhibitor of lipid peroxidation in rat liver microsomes and in phospholipid liposomes. Its cis isomer and N‐desmethyl form are weaker inhibitors, but 4‐hydroxytamoxifen is much more powerful. It is possible that the antioxidant property of tamoxifen might contribute to its biological actions.
Degradation of transcription factors, c‐Jun and c‐Fos, by calpain
Authors: Hirai Syu-ichi , Kawasaki Hiroshi , Yaniv Moshe , Suzuki Koichi
Published: July 2002

c‐Jun protein, and AP1/PEA1 transcription factor component, is a typical short‐lived protein, and like other short‐lived proteins such as c‐Fos, contains PEST regions. Calcium‐dependent neutral protease (calpain), a candidate for the degradation of PEST‐containing proteins, digests c‐Jun and c‐Fos efficiently in vitro. This is the first demonstration that transcription factors are substrates for calpain. The C‐terminal portion of c‐Jun is relatively resistant to calpain such that an 18kDa fragment, which includes the DNA binding domain, accumulates under moderate digestion conditions. The activity of c‐Jun in cultured cells can be modified by changing the level of calpastatin, an endogenous calpain inhibitor, indicating that c‐Jun is also a substrate for calpain in vivo.
Identification of the phosphorylation sites in elongation factor‐2 from rabbit reticulocytes
Authors: Price Nigel T. , Redpath Nicholas T. , Severinov Konstantin V. , Campbell David G. , Russell J.M. , Proud Christopher G.
Published: July 2002

The sites in eukaryotic elongation factor eEF‐2 phosphorylated by the Ca2+/calmodulin‐dependent eEF‐2 kinase in vitro have been identified. The kinase catalysed the rapid incorporation of one mol of phosphate per mol eEF‐2 and the slower incorporation of a second mol. All the phosphorylation sites in eEF‐2 are contained in the CNBr fragment corresponding to residues 22‐155. Tryptic digestion of phosphorylated eEF‐2 yielded 3 phosphopeptides, one being unique to monophosphorylated eEF‐2. The phosphorylation sites were identified as threonine residues 56 and 58, the former being more rapidly phosphorylated. Ala‐Gly‐Glu‐Thr‐Phe‐Thr14‐Asp‐Thr18‐Arg. The same sites are labelled in eEF‐2 isolated from reticulocyte lysates.
Two α subunits of the Gq class of G proteins stimulate phosphoinositide phospholipase C‐β1 activity
Authors: Taylor Stephen J. , Exton John H.
Published: July 2002

Two G protein α subunits were detected in preparations of GTPγS‐dependent, phosphoinositide‐specific phospholipase C‐activating proteins from bovine liver membranes. Partial resolution of the two α subunits, of molecular mass 42 and 43 kDa, was achieved by Mono Q chromatography. Quantitation of the levels of each α subunit and reconstitution assays demonstrated that each possessed stimulatory activity towards the β1 isozyme of phospholipase C. Immunoblot analysis showed that the 42 kDa protein was immunologically related to αq, whereas the 43 kDa protein was related to α11, another member of the Gq class. The data thus show that two different α subunits of the Gq class of G proteins stimulate phospholipase C‐β1 Activity.
Crystallization of the seryl‐tRNA synthetase: tRNAser complex of Escherichia coli
Authors: Price Steven , Cusack Stephen , Borel Franck , Berthet-Colominas Carmen , Leberman Reuben
Published: July 2002

Crystals of the complex between seryl‐tRNA synthetase and tRNA2
ser from Escherichia coli have been obtained from ammonium sulphate solutions. The crystals are of the 1:2 enzyme:tRNA complex, belong to the space group C222,, have cell dimensions of a = 128.9 Å, b = 164.9 Å, c = 127.3 Å and diffract anisotropically from 3.5 to 4.5 Å. An X‐ray diffraction data set to 4 Å has been collected. The combination of molecular replacement using the refined structure of the catalytic domain of the native enzyme, data from a heavy atom derivative and solvent flattening was used to produce a map at 4 Å resolution. This shows that a tRNA molecule binds across the dimer, the anticodon stem and loop do not contact the protein and the helical arm of the enzyme contacts the TΨC loop and the long extra arm of the tRNA.
Spin trapping of nitric oxide produced in vivo in septic‐shock mice
Authors: Lai Ching-San , Komarov Andrei M.
Published: July 2002

A nitric oxide (•NO) spin‐trapping technique combined with electron paramagnetic resonance (EPR) spectroscopy has been employed to measure the in vivo production of •NO in lipopolysaccharide (LPS)‐treated mice. The in vivo spin‐trapping of •NO was performed by injecting into mice a metal—chelator complex, consisting of N‐methyl‐d‐glucamine dithiocarbamate (MGD) and reduced iron (Fe2+), that binds to •NO and forms a stable, water‐soluble [(MGD)2‐Fe2+‐NO] complex, and by monitoring continuously the in vivo formation of the latter complex using an S‐band EPR spectrometer. At 6 h after intravenous injection of LPS, a three‐line EPR spectrum of the [(MGD)2‐Fe2+‐NO] complex, was observed in the blood circulation of the mouse tail; the [(MGD)2‐Fe2+] complex was injected subcutaneously 2 h before EPR measurement. No signal was detected in control groups. Administration of N
G‐monomethyl‐l‐arginine, an •NO synthase inhibitor, caused a marked reduction in the in vivo EPR signal of the [(MGD)2‐Fe2+‐NO] complex, suggesting that the •NO detected is synthesized via the arginine‐nitric oxide synthase pathway. The results presented here demonstrated, for the first time, the in vivo real time measurement of •NO in the blood circulation of conscious, LPS‐treated animals.
Apolipoprotein E expression at neuromuscular junctions in mouse, rat and human skeletal muscle
Authors: Akaaboune Mohammed , Villanova Marcello , Festoff Barry W. , Verdière-Sahuqué Martine , Hantaï Daniel
Published: July 2002

Apolipoprotein E (ApoE)‐ε4 allele has been associated with late onset familial Alzheimer's disease (AD). In both familial and sporadic AD brain, ApoE is localized to the vessel walls, senile amyloid plaques, and neurofibrillary tangles. ApoE is also an ‘injury‐response’ macromolecule in peripheral nerves and was reported to increase in response to injury. We have demonstrated that Alzheimer β‐amyloid precursor protein and a serpin α1‐antichymotrypsin also found accumulated in senile plaques in AD brain, were also localized at neuromuscular junctions (NMJs). Using immunocytochemistry, our present results indicate that ApoE is found in normal mouse, rat and human skeletal muscle and concentrated at the NMJs as it is in the senile plaques in AD brain. Such experiments may shed light on the mechanism of synapse loss, as well as plaque formation in this neurodegenerative disease.
The three dimensional structure of rat cytokine CINC/Gro in solution by homonuclear 3D NMR
Authors: Hanzawa Hiroyuki , Haruyama Hideyuki , Watanabe Kazuyoshi , Tsurufuji Susumu
Published: July 2002

The solution conformation of rat cytokine‐induced neutrophil chemoattractant (CINC/Gro), a small protein consisting of 72 amino acid residues with proinflammatory activities, and a member of the interleukin 8 family corresponding to a counterpart of human Gro, was investigated with homonuclear 2D and 3D NMR spectroscopy. At each phase of the structural analysis, the homonuclear 3D NOESY‐HOHAHA and HOHAHA‐NOESY spectra afforded valuable data, removing ambiguities intractable by conventional 2D NMR techniques. CINC/Gro exists as a dimer in solution and contains a triple stranded anti‐parallel β‐sheet and C‐terminal α‐helix in the monomer structure, as observed in human IL‐8, but non‐trivial differences are also observed.
Proteolytic modification of membrane‐associated phospholipase C‐β by μ‐calpain enhances its activation by G‐protein βγ subunits in human platelets
Authors: Banno Yoshiko , Asano Tomiko , Nozawa Yoshinori
Published: July 2002

Membrane‐associated phosphoinositide‐phospholipase C (PI‐PLC)‐β (150 kDa) and its truncated forms (100 kDa and 45 kDa) were purified from human platelets. The 100 kDa PI‐PLC‐β was found to be activated to a greater extent by brain G‐protein βγ subunits compared to the intact 150 kDa enzyme. Furthermore, treatment with μ‐calpain of the intact PI‐PLC‐β (150 kDa) caused a marked augmentation of its activation by βγ subunits. This enhanced PLC activation by βγ subunits was due to truncation by μ‐calpain, producing a 100 kDa PI‐PLC, but not by another protease,thrombin.
The oxidation of low density lipoprotein by cells or iron is inhibited by zinc
Authors: Wilkins Gary M. , Leake David S.
Published: July 2002

We have examined the effect of zinc ions on low density lipoprotein (LDL) oxidation by macrophages, endothelial cells and iron ions in terms of the increased uptake of the LDL by macrophages. Zinc ions inhibited LDL modification by both cell types (which is dependent on the presence of iron ions in the culture medium) and by iron ions alone. As oxidised LDL is believed to be involved in atherogenesis, this raises the possibility that zinc may be an endogenous protective factor against atherosclerosis.
Function of Pro‐185 in the ProCys of conserved motif IV in the EcoRII [cytosine‐C5]‐DNA methyltransferase
Authors: Kossykh Valeri G. , Schlagman Samuel L. , Hattman Stanley
Published: July 2002

ProCys in the conserved sequence motif IV of [cytosine‐C5]‐DNA methyltransferases is known to be part of the catalytic site. The Cys residue is directly involved in forming a covalent bond with the C6 of the target cytosine. We have found that substitution of Pro‐185 with either Ala or Ser resulted in a reduced rate of methyl group transfer by the EcoRII DNA methyltransferase. In addition, we observed an increase in the K
m for substrate (AdoMet), but a decrease in the K
m for substrate DNA. This is reflected in minor changes in k
cat/K
m for DNA, but in 10‐ to 100‐fold reductions in k
cat/K
m for AdoMet. This suggests that Pro‐185 is important to properly orient the activated cytosine and AdoMet for methyl group transfer by direct interaction with AdoMet and indirectly via the Cys interaction with cytosine.
Using discounted cash flow heuristics to improve project net present valve
Authors: Pinder Jonathan P. , Marucheck Ann S.
Published: July 2002

The problem of scheduling resource‐constrained projects to maximize their Net Present Value (NPV) is studied in this paper. The performance of seventeen scheduling heuristics, ten of which are original to this work, is evaluated. Heuristic performance is rated separately on maximization of project NPV and minimization of project duration. Two of the new heuristics are significantly better than the standard heuristics at yielding higher project NPVs. These two heuristics also perform well at reducing project duration. Two heuristics, Minslack and Shortest Processing Time (SPT), that performed well in previous studies are shown to be poor performers at maximizing project NPV.
TAK1 mediates an activation signal from toll‐like receptor(s) to nuclear factor‐κB in lipopolysaccharide‐stimulated macrophages
Authors: Irie Takashi , Muta Tatsushi , Takeshige Koichiro
Published: July 2002

Stimulation of monocytes/macrophages with lipopolysaccharide (LPS) results in activation of nuclear factor‐κB (NF‐κB), which plays crucial roles in regulating expression of many genes involved in the subsequent inflammatory responses. Here, we investigated roles of transforming growth factor‐β activated kinase 1 (TGF‐TAK1), a mitogen‐activated protein kinase kinase kinase (MAPKKK), in the LPS‐induced signaling cascade. A kinase‐negative mutant of TAK1 inhibited the LPS‐induced NF‐κB activation both in a macrophage‐like cell line, RAW 264.7, and in human embryonic kidney 293 cells expressing toll‐like receptor 2 or 4. Furthermore, we demonstrated that endogenous TAK1 is phosphorylated upon simulation of RAW 264.7 cells with LPS. These results indicate that TAK1 functions as a critical mediator in the LPS‐induced signaling pathway.
Isolation, reconstitution and functional characterisation of the Rhodobacter sphaeroides photoactive yellow protein
Authors: Haker Andrea , Hendriks Johnny , Gensch Thomas , Hellingwerf Klaas , Crielaard Wim
Published: July 2002

We report the isolation, functional reconstitution and photophysical characterisation of Rhodobacter sphaeroides photoactive yellow protein (PYP), of which the gene was recently cloned. Reconstitution of the his‐tagged purified apo‐protein with 4‐hydroxy‐cinnamic acid yields the characteristic blue absorbance at 446 nm, but surprisingly also an absorbance peak at 360 nm. This additional peak is not caused by binding of a second chromophore, as confirmed with mass spectroscopy. Moreover, reconstitution with the ‘locked’ analogue 7‐hydroxy‐coumarin‐3‐carboxylic acid yields only a single absorbance peak at 441 nm. The 446 nm and 360 nm species are part of a temperature‐ and pH‐dependent equilibrium. Photoactivation of the protein leads to formation of a blue‐shifted intermediate as in other PYPs, with a 100‐fold increased groundstate recovery rate (k
pB→pG=500 s−1) compared to E‐PYP.
Modulation of WHSC2 expression in human endothelial cells
Authors: Mariotti M , Manganini M , Maier J.A.M
Published: July 2002

WHSC2, a novel gene recently isolated within the critical region of Wolf–Hirschhorn syndrome, is expressed in endothelial cells. WHSC2 is downregulated by HIV‐1 Tat, whereas it is not modulated by angiogenic and pro‐differentiative molecules. WHSC2 encodes a basic polypeptide of 528 amino acids. The in vitro translated protein shows a molecular weight of 57 kDa. WHSC2 has two nuclear translocation sequences which actively mediate its transport to the nucleus, as shown in whsc2‐GFP‐transfected NIH‐3T3. We also found a helix‐loop‐helix (HLH) motif in region 130–185. Since members of the HLH family control differentiation and cell cycle progression, we hypothesize that WHSC2 may function as a transcriptional repressor.
Green fluorescent protein‐based halide indicators with improved chloride and iodide affinities
Authors: Galietta Luis J.V , Haggie Peter M , Verkman A.S
Published: July 2002

The green fluorescent protein YFP‐H148Q is sensitive to halides by a mechanism involving halide binding and a shift in pK
a. However, a limitation of YFP‐H148Q is its low halide sensitivity, with K
d>100 mM for Cl−. Indicators with improved sensitivities are needed for cell transport studies, particularly in drug discovery by high‐throughput screening, and for measurement of Cl− concentration in subcellular organelles. YFP‐H148Q libraries were generated in which pairs of residues in the vicinity of the halide binding site were randomly mutated. An automated procedure was developed to screen bacterial colonies for improved halide sensitivity. Analysis of 1536 clones revealed improved anion sensitivities with K
d down to 2 mM for I− (I152L), 40 mM for Cl− (V163S), and 10 mM for NO3
− (I152L). The anion‐sensitive mechanism of these indicators was established and their utility in cells was demonstrated using transfected cells expressing the cystic fibrosis transmembrane conductance regulator chloride channel.
Identification of Raf‐1 Ser621 kinase activity from NIH 3T3 cells as AMP‐activated protein kinase
Authors: Sprenkle Amy B , Davies Stephen P , Carling David , Hardie D.Grahame , Sturgill Thomas W
Published: July 2002

© Federation of European Biochemical Societies.
Colchicine affects protein kinase C‐induced modulation of synaptic transmission in cultured hippocampal pyramidal cells
Authors: Bouron Alexandre
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Formation of a 2‐methyl‐branched fatty aldehyde during peroxisomal α‐oxidation
Authors: Croes Kathleen , Casteels Minne , Asselberghs Stanny , Herdewijn Piet , Mannaerts Guy P , Van Veldhoven Paul P
Published: July 2002

In the final reaction of peroxisomal α‐oxidation of 3‐methyl‐branched fatty acids a 2‐hydroxy‐3‐methylacyl‐CoA intermediate is cleaved to formyl‐CoA and a hitherto unidentified product. The release of formyl‐CoA suggests that the unidentified product may be a fatty aldehyde. When purified rat liver peroxisomes were incubated with 2‐hydroxy‐3‐methylhexadecanoyl‐CoA 2‐methylpentadecanal was indeed formed. The production rates of formyl‐CoA (measured as formate) and of the aldehyde were in the same range. While the production of formate remained unaltered in the presence of NAD+, the amount of 2‐methylpentadecanal was decreased, which was accompanied by the formation of 2‐methylpentadecanoic acid. These data indicate that (1) during α‐oxidation the 2‐hydroxy‐3‐methylacyl‐CoA is cleaved to a 2‐methyl‐branched aldehyde and formyl‐CoA and (2) liver peroxisomes are capable of converting this aldehyde to a 2‐methyl‐branched fatty acid.
Only the last amino acids in the nascent peptide influence translation termination in Escherichia coli genes
Authors: Mottagui-Tabar Salim , Isaksson Leif A
Published: July 2002

Efficiency of translation termination is affected if the last two amino acids in the nascent peptide are changed [1, 2]. By changing the corresponding codons upstream of the stop signal UGAA, we have analyzed if the −3 to −6 amino acids at the C‐terminal region of the nascent peptide also affect termination. Lysine at position −3 gave increased readthrough, whereas a total of 28 variations at positions −4, −5, and −6 showed no significant effect on readthrough. The 3′‐ends corresponding to the last six codons in 27 Escherichia coli genes were inserted upstream of a stop codon in the 3A′ translation assay gene [1]. Readthrough of the stop codon was measured and a possible correlation with the Codon Adaptation Index (CAI) [3]of the respective genes was investigated. Sequences from genes with low CAI do not give any such correlation, whereas sequences from genes with high CAI values are correlated with high termination efficiency. This correlation disappears if the −1 and −2 codons/amino acids are changed. The results suggest that mainly the terminal dipeptide of the terminal hexapeptide sequence has an influence on termination in the tested E. coli genes. This influence is dependent on the charge of the −2 amino acid and is correlated with the α‐helix propensity of the −1 amino acid, in accordance with results obtained from synthetic gene constructs [1, 2].
Oxygen dependence of photosynthetic electron transport in a bacteriochlorophyll‐containing rhizobium
Authors: Kramer David M , Kanazawa Atsuko , Fleischman Darrell
Published: July 2002

Bacteriochlorophyll‐containing rhizobia, which form nitrogen‐fixing nodules on the stems and roots of the legume Aeschynomene, grow photosynthetically only in the presence of oxygen or auxiliary electron acceptors. We show that, in whole cells of the Rhizobium strain BTAi 1, a single‐turnover excitation flash photooxidized c‐type cytochrome under aerobic but not anaerobic conditions. Light‐induced fluorescence yield changes show that under anaerobic conditions, the primary acceptor quinone, QA, is predominantly in the reduced state and so unable to accept electrons. Thus, as is the case for the aerobic photosynthetic bacterium Roseobacter denitrificans, over‐reduction of QA likely prohibits photosynthesis under anaerobic conditions.
Alternative promoter usage and tissue specific expression of the mouse somatostatin receptor 2 gene
Authors: Kraus Jürgen , Wöltje Michael , Schönwetter Nadja , Höllt Volker
Published: July 2002

We have cloned the 5′ upstream regulatory region of the mouse somatostatin receptor 2 gene. Its genomic organization is novel among all somatostatin receptor genes. It contains two previously unrecognized exons, separated by introns larger than 25 kb, and three tissue and cell specific alternative promoters. The first promoter in front of exon 1 is active only in AtT‐20 tumor cells. The second promoter, located 5′ to exon 2, is used in brain, pituitary, adrenals, pancreas, NG 108‐15 and AtT‐20 cells. Furthermore, it contains putative DNA elements for regulation by glucocorticoids, estradiol and cAMP. A third promoter, located in exon 3, is additionally used in lung, kidney and spleen.
Short‐term effects of leptin on lipid metabolism in the rat
Authors: López-Soriano Joaquı́n , Carbó Neus , López-Soriano Francisco J , Argilés Josep M
Published: July 2002

In this study, we have examined the short‐term effects of leptin on lipid metabolism in the rat. Acute leptin administration induced hypertriglyceridaemia (31% increase in plasma triacylglycerols) which was not associated with changes in lipoprotein lipase activity in white adipose tissue. Surprisingly, leptin administration did not induce any changes in the lipogenic rate in either white adipose tissue or liver. Leptin administration caused a decreased tissue uptake of exogenous 14C‐triacylglycerols. These data suggest that leptin induces important changes in lipid uptake in adipose tissue and skeletal muscle which could be responsible for the observed hypertriglyceridaemia.
Neuronal expression of fractalkine in the presence and absence of inflammation
Authors: Schwaeble Wilhelm J , Stover Cordula M , Schall Thomas J , Dairaghi Daniel J , Trinder Peter K.E , Linington Christopher , Iglesias Antonio , Schubart Anna , Lynch Nicholas J , Weihe Eberhard , Schäfer Martin K.-H
Published: July 2002

Fractalkine is the only as yet known member of a novel class of chemokines. Besides its novel Cys‐X‐X‐X‐Cys motif, fractalkine exhibits features which have not been described for any other member of the chemokine family, including its unusual size (397 amino acids human, 395 mouse) and the possession of a transmembrane anchor, from which a soluble form may be released by extracellular cleavage. This report demonstrates the abundant mRNA and fractalkine protein expression in neuronal cells. The neuronal expression of fractalkine mRNA is unaffected by experimentally induced inflammation of central nervous tissue.