bluuebunny/crossref_metadata_2025 huggingface dataset:
Structural Assessment Considering Architectural Constraints with Seismic Retrofitting
Authors: Sanoufi Abdulrahman , Bawazeer Hani , Sebaei Yoseff , Shaheen Yousef , Naqash Muhammad Tayyab
Published: July 2021

The Kingdom of Saudi Arabia is located in a low to medium earthquake zone. Therefore, the inclusion of seismic loads in building design was limited to specific building structures such as highrise and midrise in the past decades. Recently, the development and adoption of the Saudi Building Code (SBC) and the experienced seismic activity at many regions in the Kingdom necessitate detailed seismic design considerations for all buildings. Given this, the current work initially emphasizes assessing structural grids obtained from an architectural plan for an existing building in AL Madina. Then the structure has been analyzed critically in such a way to reduce columns and simplify the structural grid. Also, the orientation of columns has been modified to obtain structure symmetry keeping in view the architectural constraints. Two cases have been developed initially: flat slab and solid slab and designed to withstand gravity loads using Saudi Building Provisions. These cases are analyzed for the seismicity of the Medina Region. Since Medina is less prone to seismicity, the building withstands the lateral load calculated based on static analysis. To assess these buildings for stronger earthquakes, we increased the applied load to assess their capacity. Since both the proposed cases fail to withstand the increased seismic load, a bracing system has been introduced at the locations where it does not disturb the architecture of the building. It was observed that introducing bracing improves the performance of the structures. Therefore concluding that complex structural grids schemes can be simplified, regularized, and economized as well. In addition, bracings provide an easy technique to retrofit already existing RC buildings.
Potable Water Scarcity and Options for Effective Provision in Limbe Municipality, Southwest Region, Cameroon
Authors: Buh Emmanuel Nong , Mbua Roy Lyonga , Ngong Ukah Bonaventure
Published: July 2021

Continuous potable water scarcities in Limbe municipality if not well manage and ensure a regular provision of it may lead to many disastrous consequences to the massive growing population of this municipality. Lack of safe drinking water is a severe crisis in Limbe and Cameroon due to poor management strategies employed by the water management bodies involved. The study investigated the implications of water scarcity in Limbe municipality and the options for effective provision of potable water. A questionnaire survey was carried out. About 300 questionnaires were administered in three localities of Limbe to investigate the present status of water scarcity and its impacts on the local communities. The results show an increasing rate of potable water scarcity in the municipality. About 51% of the respondents indicated that inadequate storage facilities are the main caused of water scarcity in the town. In contrast, 26% and 23% of the respondents indicated that inadequate technology and low level of education are other reasons contributing to potable water scarcity in Limbe municipality. Many people in the municipality depend on untreated water sources, resulting in the contraction of many diseases like dysentery and diarrhea as prime waterborne health risks. We suggest options like installing large water safety tanks in all the localities of Limbe municipality. Furthermore, implementing a joint monitoring team to ensure that effective potable water treatment and supply is being done and the bodies involved in water management like the Cameroon Water Utilities Corporation (CAMWATER) need to initiate a platform for multi-stakeholder dialogue, networking and social learning essentially for building trust on how to manage potable water supply in the municipality better.
Analysis of Blood Serum Bilirubin Concentration among Moderate Intensity Exercise Performers; A Randomized Control Trails
Authors: Khan Adnan , Khan Alamgir , Butt Muhammad Zafar Iqbal , Khan Shahzaman , Khan Samiullah , Selamoglu Zeliha , Jamil Muhammad , Manan Abdul
Published: July 2021

Purpose: A randomized control trails were conducted to evaluate the effects of moderate intensity exercise on blood serum bilirubin level.

Methods:  A participants of the study were divided in two groups i.e. experimental group (EG= 10) and control group (CG=10).  5ml blood were taken from all the subjects and each blood sample was marked with separate identification code. Twelve weeks exercise (moderate intensity exercise) protocols was employed on the selected subjects. The collected blood samples were tested and thus the collected results were processed through Statistical Package for Social Sciences (SPSS, Version, 24) by applying Mean and Standard deviation etc.

Result: Based on analysis the researcher concluded that moderate intensity exercise has significant impact on Serum Bilirubin Concentration of the subjects. In addition significant difference was found in pre and post test result of experimental group  (t4= 1.000, Sig. = .374 > α= 0.05).
COVID-19 Mutation and Impact on Various Organs
Authors: Chintalwar Kunal , Chakole Swaroopa
Published: July 2021

Background: COVID-19 is the continuously evolving pandemic affecting billions of people. The viruses are notorious for mutations in them. Novel coronavirus is no exception to mutation and mutated into various strains according to geographical locations.

Summary: The recent second and third wave which is actually a resurgence in COVID-19 cases after a steep fall is attributed to the mutation that is occurring in novel coronavirus. Various strains has been isolated and is being under study to tackle their menace. But blaming completely the mutant version of the virus is unjust as there are other factors such as lax implementation of preventive measures, negligence on account of people by not wearing masks and not following physical distancing. The impact of COVID-19 on human organ system is almost negative and affects the functioning of the organ adversely. Therefore proper care must be taken to ensure the safety of oneself.

Conclusion: Mutations in viruses cannot be stopped and solution lies within us. By following preventive measures and guidelines issued by WHO and other competent authority can a key to keep oneself self. Also balance diet along with proper exercise can help in these pandemic. More study needs to be done to establish various correlation so than containment plan of COVID-19 can be modifies accordingly.
Cannabis (Cannabis sativa) Containing Drug Formulations Mentioned in “Thalpathe piliyam”: A Review
Authors: Diddeniya J. I. D. , Kulatunga W. M. S. S. K.
Published: July 2021

Introduction: Cannabis sativa is an herbaceous plant which is used as a remedy for many diseases in Traditional Medicine of Sri Lanka. Even though cannabis is the most illicit plant around the world, its medicinal properties are beneficial in numerous ways. The active ingredients of cannabis are Cannabinol, Cannabidiol, Tetrahydro cannabinol, Tetrahydro cannabivarin etc.

Objectives: The aim of this study was to make an overview of the different formulae containing cannabis, their system wise indications, and preparatory methods and to analyze the amount of cannabis percentage in the formulations mentioned in “Thalpathe Piliyam”. And also to identify the social economic impact of cannabis to the community through this.

Methodology: For this review all the twenty one “Thalpathe Piliyam” books were used and other necessary facts were obtained from published research articles in data bases such as Science direct, PubMed etc.

Results: The results revealed that, majority of drug formulations, that is 49.23% acts on the Anna pachana Vyuhava (Gastro intestinal tract) and apart from that 40.77% acts on Naadi vyuhava (Nervous system), 7.69% on Raktha vyuhava (Circulatory system), 6.54% on Prajanana sansthaana (Reproductive system), 5.77% on Swasana sansthaana (Respiratory system) and 22.69% on other diseases by giving relief to diseases such as indigestion, vomiting, cancer, diabetes, piles, cough, diarrhea etc. Among the different preparatory methods of drug formulations which contain Cannabis as an ingredient, “Guli” (pills) was the most abundant formulation with a percentage of 48.07%. Other formulations such as Kalka (pastes), Kashaya (decocotion), Swarasa (fresh juice), Paththu (poultice) were also mentioned in the book. By analyzing the amount of Cannabis percentage, the highest amount of the drug preparations were in between the range of less than or equal to 5% while the least amount of drug preparations were in the range from more than 20%. This indicates that most preparations contain Cannabis less than 5% as an amount in them.

Conclusion: Therefore it can be concluded that, “Thalpathe Piliyam” is a rich repository for the valuable traditional formulae which consist of multiple formulations which contains Cannabis and also very small amount of Cannabis amount can act very efficiently in curing diseases. Furthermore, its socio economic impact to the community can be also measured through it.
Do Energy Poverty and Energy Consumption Drive CO2 Emissions? Evidence from Sub-Saharan Africa
Authors: Juliette Mewamba-Chekem , Willy Noumessi Fodjou
Published: July 2021

Energy poverty is a multidimensional concept, but in the developing country context, it could refer to the lack or difficulty of people to access modern and reliable energy services. It refers particularly to access to electricity and to modern and clean cooking fuels. This limited access constitutes a serious hindrance to socioeconomic development and has adverse consequences on environment. The present study attempts to investigation the contribution of energy poverty and consumption to CO2 emissions in 20 Sub-Saharan Africa countries over 1996-2015. Using Feasible Generalized Least Squares (FGLS) and Panel-corrected standard errors (PCSE), our results broadly suggest non-significant effect of energy poverty on CO2 emissions. However, we found evidence that primary energy consumption drives CO2 emissions. To mitigate the adverse effects of energy consumption on environment, governments should design and implement policies to improve energy efficiency and promote renewable energies use.
A New Approach for the Green Biosynthesis of Silver Oxide Nanoparticles Ag2O, Characterization and Catalytic Application
Authors: El-Ghmari Brahim , Farah Hanane , Ech-Chahad Abdellah
Published: July 2021

In this paper, a facile and green approach for the synthesis of silver oxide nanoparticles Ag2O NPs was performed using the extract of the wild plant Herniaria hirsuta (H. hirsuta). Different spectral methods were used for the characterization of the biosynthesized Ag2O NPs, ultraviolet-visible (UV-Vis) spectroscopy gave a surface plasmon resonance (SPR) peak of Ag2O NPs is 430 nm, estimation of direct and indirect forbidden gap bands are respectively 3.76 eV and 3.68 eV; Fourier transform infrared (FTIR) spectral analysis revealed the groups responsible for the stability and synthesis of Ag2O NPs. The morphology of Ag2O NPs was studied by scanning electron microscopy (SEM) showing a nearly spherical shape of Ag2O NPs, and X-ray diffraction (XRD) study confirmed the crystallinity of Ag2O NPs with a crystallinity size of 15.51 nm. The catalytic activity of Ag2O NPs, as well as the rings number were studied by the degradation of methylene blue dye. Copyright © 2021 by Authors, Published by BCREC Group. This is an open access article under the CC BY-SA License (https://creativecommons.org/licenses/by-sa/4.0).
Effect of adenosine deaminase and an adenosine analogue on insulin sensitivity in soleus muscle of the rat
Authors: Espinal Joseph , John Challiss R.A. , Newsholme Eric A.
Published: July 2002

The concentration of insulin that produces half‐maximal stimulation of glycolysis in stripped soleus muscle preparations is decreased from ∼100−10 μunits/ml by the presence of adenosine deaminase in the incubation medium. This suggests that adenosine decreases insulin sensitivity. The effect of the deaminase is abolished by addition of the adenosine analogue, N
6‐phenylisopropyladenosine which is not metabolised by the deaminase. The effect of the deaminase in isolated soleus muscle is similar to that of a period of physical training of the rat.
Purification of the active mitochondrial tricarboxylate carrier by hydroxylapatite chromatography
Authors: Stipani I. , Palmieri F.
Published: July 2002

The mitochondrial tricarboxylate carrier has been extracted from rat liver mitochondria or SMP with Triton X‐100, in the presence of 1,2,3‐BTA and DPG, and partially purified by chromatography on HTP. The purified fraction, which also contains the ADP/ATP carrier and the phosphate carrier, after incorporation into liposomes catalyzes a 1,2,3‐BTA‐sensitive [14C]citrate/citrate exchange. The tissue and substrate specificity, the inhibitor sensitivity and the kinetic properties of citrate transport in liposomes are similar to those described for the citrate transport in mitochondria. The maximal rate of citrate exchange in the reconstituted system is 338 μmol·min−1·g protein−1, at 30°C and pH 7.0.
Active transport of lead by human red blood cells
Authors: Simons T.J.B.
Published: July 2002

Human red cells suspended in lead‐citrate buffers (2.6μM Pb2+) take up much less Pb than predicted from studies of equilibrium binding of Pb to haemolysates. Pb uptake is increased by ATP depletion, or by loading at 0°C. Tracer studies with 203Pb indicate that the low uptake at 37°C in the presence of substrate is not due to membrane impermeability to Pb. Cold‐loaded cells extrude Pb against a concentration gradient at 37°C when glucose is present. These results suggest that the cellular loading of Pb is dependent on the balance between an inward leak and an outward pump. The extrusion of Pb from the cells is possibly brought by the Ca pump.
Plasma gelsolin caps and severs actin filaments
Authors: Harris Harriet E. , Weeds Alan G.
Published: July 2002

Actin capping protein
Actin polymerization
Gold labeling
Role of the 33‐kDa polypeptide in preserving Mn in the photosynthetic oxygen‐evolution system and its replacement by chloride ions
Authors: Miyao Mitsue , Murata Norio
Published: July 2002

Treatment of Photosystem II particles from spinach chloroplasts with Triton X‐100 with 2.6 M urea in the presence of 200 mM NaCl removed 3 polypeptides of 33 kDa, 24 kDa and 18 kDa, but left Mn bound to the particles. The (urea + NaCl)‐treated particles could evolve oxygen in 200 mM, but not in 10 mM NaCl. Mn was gradually released with concomitant loss of oxygen‐evolution activity in 10 mM NaCl but not in 200 mM Cl−. The NaCl‐treated particles, which contained Mn and the 33‐kDa polypeptide but not the 24‐kDa and 18‐kDa polypeptides, did not lose Mn or oxygen‐evolution activity in 10 mM NaCl. These observations suggest that the 33‐kDa polypeptide maintains the binding of Mn to the oxygen‐evolution system and can be functionally replaced by 200 mM Cl−.
Binding of transferrin‐iron by adriamycin at acidic pH
Authors: Demant Erland J.F. , Nørskov-Lauritsen Niels
Published: July 2002

It is shown that adriamycin is able to chelate iron released from iron‐loaded serum transferrin in the pH range from 6.5‐4.1. The kinetics of iron transfer to free adriamycin and to adriamycin covalently attached to the transferrin has been determined. The results show that adriamycin, if introduced into intracellular acidic compartments, could function as acceptor for transferrin‐iron.
Strong orientational ordering of the near‐infrared transition moment vectors of light‐harvesting antenna bacterioviridin in chromatophores of the green photosynthetic bacterium Chlorobium limicola
Authors: Fetisova Z.G. , Kharchenko S.G. , Abdourakhmanov I.A.
Published: July 2002

The direction of the transition moments of chlorosome pigments in chromatophores of the green photosynthetic bacterium Chlorobium limicola was studied by linear dichroism. Orientation of chromatophores was achieved by stretching a polyacrylamide gel in which they were packed. It was shown that in each individual chromatophore the Qy transition moment vectors of the whole chlorosome bacterioviridin are parallel to each other and are practically ideally oriented along the chlorosome long axis. The exact value of the angle α between the bacterioviridin transition moments and the long axis of the chlorosome is calculated to be α = 0°, the mean square deviation being 7°.
Ca2+ influx mediated through the GPIIb/IIIa complex during platelet activation
Authors: Yamaguchi A. , Yamamoto N. , Kitagawa H. , Tanoue K. , Yamazaki H.
Published: July 2002

When aequorin‐loaded platelets were stimulated with thrombin, the luminescence signal of aequorin showed two peaks. From experiments with 1 mM external Ca2+ or EGTA, both one‐half of the first peak and the entire second peak reflected the influx of Ca2+ from the external medium, and the remaining half of the first peak reflected the mobilization of Ca2+ from its storage site. A monoclonal antibody (TM83) that recognizes the glycoprotein IIb/IIIa (GPIIb/IIIa) complex which has binding sites for fibrinogen and the synthetic peptide GRGDSP are known to inhibit fibrinogen binding and platelet aggregation. Both eliminated the second peak of intracellular free calcium ([Ca2+]i). Similar effects were observed during activation by collagen, but not during PMA activation. It was concluded that the GPIIb/IIIa complex was intimately related to a part of the Ca2+ influx during the activation of platelets.
Cloning and sequence analysis of the human brain β‐adrenergic receptor
Authors: Chung Fu-Zon , Lentes K.-Ulrich , Gocayne Jeannine , Fitzgerald Michael , Robinson Doreen , Kerlavage Anthony R. , Fraser Claire M. , Venter J.Craig
Published: July 2002

Two cDNA clones, λ‐CLFV‐108 and λ‐CLFV‐119, encoding for the β‐adrenergic receptor, have been isolated from a human brain stem cDNA library. One human genomic clone, LCV‐517 (20 kb), was characterized by restriction mapping and partial sequencing. The human brain β‐receptor consists of 413 amino acids with a calculated M
r of 46 480. The gene contains three potential glucocorticoid receptor‐binding sites. The β‐receptor expressed in human brain was homology with rodent (88%) and avian (52%) β‐receptors and with porcine muscarinic cholinergic receptors (31%), supporting our proposal [(1984) Proc. Natl. Acad. Sci USA 81, 272‐276] that adrenergic and muscarinic cholinergic receptors are structurally related. This represents the first cloning of a neurotransmitter receptor gene from human brain.
Temperature dependence of P680+ reduction in O2‐evolving PS II membrane fragments at different redox states Si of the water oxidizing system
Authors: Eckert H.-J. , Renger G.
Published: July 2002

The electron transfer kinetics from Z to P680+ was analyzed as a function of temperature in the range of 248<T< 295 K by measuring absorption changes induced at 830 nm by a laser flash train in dark adapted O2 evolving PS II membrane fragments from spinach. It was found: (i) that the kinetics of P680+ reduction and their dependence on the redox state S
i
of the catalytic site of water oxidation are only slightly affected by temperature within the physiological range of 270<T<295 K. (ii) In the dark relaxed state S, the electron transfer from Z to P680+ exhibits an activation energy of the order of 10 kJ/mol in 248<T<295 K. (iii) In the 2nd and subsequent flashes of the train the ability for a stable charge separation between P680+ and Q−
A, markedly decreases below −10°C. This phenomenon is assumed to be due to a strong effect of temperature on the electron transfer from Q−
A to QB. The results are briefly discussed in relation to possible effects of structural changes in the D‐1/D‐2 polypeptide complex on the reaction coordinate of electron transfer steps in PS II.
Crosslinking of ribosomal protein S18 to 16 S RNA in E. coli ribosomal 30 S subunits by the use of a reversible crosslinking agent: Trans‐diamminedichloroplatinum(II)
Authors: Moine Hervé , Bienaimé Christian , Mougel Marylène , Reinbolt Joseph , Ebel Jean Pierre , Ehresmann Chantal , Ehresmann Bernard
Published: July 2002

We have previously developed [(1987) Biochemistry 26, 5200–5208] the use of trans‐diamminedichloroplatinum(II) to induce reversible RNA‐protein crosslinks in the ribosomal 30 S subunit. Protein S18 and, to a lesser extent, proteins S13/S14, S11, S4 and S3 could be crosslinked to the 16 S rRNA. The aim of the present work was to identify the crosslinking sites of protein S18. Three sites could be detected: a major one located in region 825–858, and two others located in regions 434–500 and 233–297. This result is discussed in the light of current knowledge of the topographical localization of S18 in the 30 S subunit and of its relation with function.
Evidence for lipoxin formation by bovine polymorphonuclear leukocytes via triple dioxygenation of arachidonic acid
Authors: Walstra Pauline , Verhagen Jan , Vermeer Mario A. , Klerks Johannes P.M. , Veldink Gerrit A. , Vliegenthart Johannes F.G.
Published: July 2002

Incubation of bovine polymorphonuclear leukocytes (PMNs) with arachidonic acid leads to the formation of four lipoxins. The same lipoxins are also formed upon incubation of bovine PMNs with 5(S)‐hydroperoxy‐6‐trans‐8,11,14‐cis‐eicosatetraenoic acid, 5‐hydroxy‐6‐trans‐8,11,14‐cis‐eicosatetraenoic acid, 5(S)‐hydroperoxy, 15(S)‐hydroxy‐6,13‐trans‐8, 11‐cis‐eicosatetraenoic acid or 5(S), 15(S)‐dihydroxy‐6,13‐trans‐8,11‐cis‐eicosatetraenoic acid. A 5,6‐epoxide as intermediate in lipoxin formation in the bovine PMN is highly improbable because the 5‐hydroxy compounds are as good substrates as the 5‐hydroperoxy compounds. Moreover, the two main lipoxins were found to coelute with the two lipoxins produced via a triple dioxygenation of arachidonic acid by soybean lipoxygenase‐1. Hence the bovine PMN is the first cell for which evidence is presented that the formation of lipoxins proceeds mainly via triple dioxygenation and not via 15‐hydroxy‐leukotriene A4 as is proposed for human and porcine PMNS.
Stabilisation and immobilisation of penicillin amidase
Authors: Burteau N. , Burton S. , Crichton R.R.
Published: July 2002

Penicillin amidase was coupled to a periodate‐oxidised dextran by reductive alkylation in the presence of sodium cyanoborohydride. A loss of activity (25%) was observed but the conjugate enzyme dextran was more thermostable than the native enzyme. Native and dextran‐conjugated penicillin amidase were immobilised on amino activated silica (Promaxon, Spherosil, Aerosil) by a classical method using glutaraldehyde for the native enzyme and reductive alkylation for the modified enzyme. Good relative activity of the enzymes was obtained after insolubilisation. Immobilisation of both native and modified enzymes resulted in the thermostabilisation of the penicillin amidase.
Interactions of papaya proteinase IV with inhibitors
Authors: Buttle David J. , Ritonja Anka , Dando Pamela M. , Abrahamson Magnus , Shaw Elliot N. , Wikstrom Peter , Turk Vito , Barrett Alan J.
Published: July 2002

Papaya proteinase IV (PPIV) is not inhibited by chicken cystatin, or human cystatins A or C, unlike most other proteinases of the papain superfamily. The enzyme inactivates chicken cystatin and human cystatin C by limited proteolysis of the glycyl bond previously shown to be involved in the inhibitory inactivity of the cystatins, but has no action on cystatin A. Contamination of commercial crystalline papain with PPIV accounts for the limited proteolysis of cystatins by ‘papain’ reported previously. PPIV is slowly bound by human α2‐macroglobulin. The enzyme is irreversibly inactivated by E‐64, and by peptidyl diazomethanes containing glycine in P1 and a hydrophobic side‐chain in P2. The reaction of PPIV with iodoacetate is extremely slow. PPIV is inhibited by peptide aldehydes despite the presence of bulky sidechains in P1, suggesting that these reversible inhibitors do not bind as substrate analogues.
Evolutionary origin of numerous kringles in human and simian apolipoprotein(a)
Authors: Ikeo Kazuho , Takahashi Kei , Gojobori Takashi
Published: July 2002

Human apolipoprotein(a) has a great size heterogeneity and consists of 38 kringle domains in the amino terminal and a serine proteasc domain in the carboxyl terminal. All but one kringle of apolipoprotein(a) are homologous to the fourth kringle of plasminogen. However, the 38th kringle resembles the fifth kringle of plasminogen and it seems to have been deleted in simian species. The phylogenetic trees suggest that an ancestral apolipoprotein(a) may have started with a duplicate of a plasminogen type protein. It also implies that deletion of the three kringles in the amino terminus followed, and that one of the remaining two kringles was duplicated in both human and simian species and the other was processed by a deletion in simian species after species separation. Thus, the number of kringles in other mammals not yet studied may vary considerably from species to species.
DNA‐specific antiidiotypic antibodies in the sera of patients with autoimmune diseases
Authors: Bronshtein Igor B. , Shuster Alexander M. , Gololobov Gennady V. , Gromova Irina I. , Kvashuk Oksana A. , Belostotskaya Ksenia M. , Alekberova Zemfira S. , Prokaeva Tatiana B. , Gabibov Alexander G.
Published: July 2002

Blood sera of patients with autoimmune diseases scleroderma (Scl), systemic lupus erythematosus (SLE), and rheumatoid arthritis (RA) have been shown to yield a specific immune response to topoisomerase I, the product of expression of a cDNA fragment cloned into λgt11 and monoclonal antibodies (MAB) to the enzyme. The ‘topoisomerase test’ is not absolutely specific for Scl. The stable positive response of autoimmune sera to anti‐topoisomerase monoclonal antibodies has a specific character and is associated with the interaction of the Fab fragment of MAB with the IgG fraction of autoimmune serum. The response observed indicates the induction of anti‐idiotypic antibodies against topoisomerase. The anti‐idiotype, isolated by HPLC and affinity chromatography demonstrated the following functional activities: (i) the immunological reaction against DNA; (ii) high‐affinity DNA‐binding with topoisomerase‐specific consensus; (iii) ability to compete with the native enzyme for binding with DNA and MAB to topoisomerase; (iv) immunological reaction against MAB to topoisomerase.
Assay conditions for the mitochondrial NADH:coenzyme Q oxidoreductase
Authors: Estornell Ernesto , Fato Romana , Pallotti Francesco , Lenaz Giorgio
Published: July 2002

The assay of Complex I activity requires the use of artificial acceptors, such as short‐chain coenzyme Q homologs and analogs, because the physiological quinones, such as CoQ10, are too insoluble in water to be added as substrates to the assay media. The medical interest raised in the last years on the pathological changes of Complex I activity has focussed on the requirement of easy reliable assays for its analysis. We have undertaken a systematic examination of the assay conditions of Complex I in mitochondrial membranes, using a series of quinones as electron acceptors, particularly the coenzyme Q homologs CoQ0, CoQ1 and CoQ2, and the analogs duroquinone and decylubiquinone. Our findings have pointed out that the most suitable electron acceptor for the NADH:CoQ reductase assay is the homolog CoQ1. The analog DB, commercially available, although yielding a high activity, nevertheless causes some problems for the standardization of the assay conditions.
Molecular action of the type 1 ribosome‐inactivating protein saporin 5 on Vicia sativa ribosomes
Authors: Iglesias Rosario , Arias F.Javier , Rojo M.Angeles , Escarmis Cristina , Ferreras J.Miguel , Girbés Tomás
Published: July 2002

The type 1 ribosome‐inactivating protein (RIP) saporin 5 isolated from seeds of Saponaria officinalis L. strongly inhibited translation carried out by Vicia sativa L. purified ribosomes. The toxin multidepurinated V. sativa rRNA, which upon treatment with acid aniline releases several RNA fragments including an RNA fragment of approximately 370 nucleotides the 5'‐end sequence of which was 5'‐GAGGAACG‐3'.
Molecular cloning of a human β3‐adrenergic receptor cDNA
Authors: Lelias J.M. , Kaghad M. , Rodriguez M. , Chalon P. , Bonnin J. , Dupre I. , Delpech B. , Bensaid M. , LeFur G. , Ferrara P. , Caput D.
Published: July 2002

We report the molecular cloning of a β3‐adrenergic receptor β3‐AR cDNA from human brown adipose tissue. The cDNA‐encoded protein is identical to the previously cloned β3‐AR but with 6 additional amino acids at the C‐terminus. The C‐terminus is shared by the β3 receptors expressed in human neuroblastoma cells [SK‐N‐MC] [Mol. Pharmacol. 42 (1992) 964‐970]. Furthermore, using a polymerase chain reaction strategy we have cloned and sequenced the β3‐AR introns. Sequence analysis demonstrates that the human β3‐AR gene comprises at least 3 exons and 2 introns and that the most abundant β3‐AR transcripts encode a protein with an exon 3‐derived C‐terminus. Interestingly, although a similar organization has been found in rodent genes, the rat β3‐AR transcripts encode a receptor with an exon 2‐derived C‐terminus.
Anion‐mediated lysine—arginine interaction evidence in Chaetopterus variopedatus sperm protamine
Authors: Piscopo Marina , Tomei Licia , Petrocellis Luciano De , Geraci Giuseppe
Published: July 2002

Chaetopterus ariopedatus sperm protamine is a stable oligomer. Specific amino acid side chain modifications show that the oligomeric structure depends on anion‐mediated lysine—arginine interactions. The occurrence of this type of interaction is confirmed by the finding that Poly‐l‐arginine readily forms aggregates with Poly‐l‐lysine or with the native but not with the protamine with carbamylated ε‐amino groups.
Signal transduction of the GLP‐1‐receptor cloned from a human insulinoma
Authors: van Eyll Brigitte , Lankat-Buttgereit Brigitte , Bode Hans P. , Göke Rüdiger , Göke Burkhard
Published: July 2002

GLP‐1 (glucagon‐like peptide 1 (7–36) amide) plays an important role in the regulation of insulin secretion and proinsulin gene expression of pancreatic β‐cells. Patients with insulinoma tumors show uncontrolled insulin hypersecretion. This study demonstrates the molecular cloning of a cDNA for the GLP‐1 receptor from a human insulinoma employing a λ‐gt 11 cDNA library. The cloned cDNA encoded a seven transmembrane domain protein of 463 amino acids which showed high homology to the GLP‐1 receptor in normal human pancreas. Four amino acid exchanges were found in comparison to a receptor sequence obtained from regular pancreatic islets. When transfected transiently into COS‐7 or stably into fibroblast CHL cells a high affinity receptor was expressed which coupled to the adenylate cyclase with normal basal cAMP and increasing intracellular cAMP levels under GLP‐1 stimulation. The receptor accepted GLP‐1 and the non‐mammalian agonist exendin‐4 as high affinity ligands. In transfected COS‐7 cells, GLP‐1 did not influence intracellular calcium, whereas in the stably transfected fibroblasts GLP‐1 transiently increased intracellular calcium to a small extent. The understanding of GLP‐1 receptor regulation and signal transduction will aid in the discovery of compounds that act as agonists of the GLP‐1 receptor for potential use in the treatment of diabetes and will facilitate the understanding of its expression under normal and pathophysiological conditions.
The polytopic mitochondrial inner membrane proteins MIM17 and MIM23 operate at the same preprotein import site
Authors: Kübrich Michael , Keil Petra , Rassow Joachim , Dekker Peter J.T. , Blom Jolanda , Meijer Michiel , Pfanner Nikolaus
Published: July 2002

Three proteins of the mitochondrial inner membrane are known that are essential for the viability of yeast and seem to be involved in import of preproteins; the integral membrane proteins MIM17 and MIM23 and the peripheral membrane protein MIM44, MIMI7 and MIM23 are homologous to each other in their hydrophobic domain, expose their termini to the intermembrane space, and span the inner membrane up to four times, each. A preprotein in transit across the mitochondrial membrane is specifically cross‐linked to MIM17, MIM23, MIM44, and matrix hsp70. We conclude that MIM17 and MIM23 are integral parts of a preprotein translocation channel and cooperate with MIM44 and hsp70 at the same protein import site.
Generation of active oxygen in elicited cells of Arabidopsis thaliana is mediated by a NADPH oxidase‐like enzyme
Authors: Desikan Radhika , Hancock John T. , Coffey Marcus J. , Neill Steven J.
Published: July 2002

Suspension‐cultured cells of Arabidopsis thaliana generated active oxygen species (AOS) (measured by luminol‐dependent chemiluminescence) following challenge with the bacterial protein elicitor harpin or the protein kinase activator phorbol 12‐myristate 13‐acetate. These responses were blocked by inhibitors of superoxide dismutase (SOD), NADPH oxidase and protein kinase. Harpin treatment also resulted in an increase in cell death, a response reduced by inhibitors of AOS generation or AOS scavengers. Extracellular SOD activity was found to be present in cell culture medium. Immunoblotting of Arabidopsis extracts revealed the presence of proteins immunologically related to the human neutrophil NADPH oxidase comples, and cell‐free reconstitution assays showed that human neutrophil cytosol combined with Arabidopsis membranes could initiate superoxide generation. These data suggest that the enzyme catalysing the generation of superoxide in elicited Arabidopsis cells is similar to the mammalian NADPH oxidase and that a signalling cascade leading to AOS generation involves protein phosphorylation.
Decreasing rates of incident schizophrenia cases in psychiatric service: a review of the literature
Authors: Munk-Jørgensen P
Published: July 2002

A review of the literature shows that the admission rates of new cases of schizophrenia vary with a factor of two to five in North American studies as well as in European studies. Furthermore, the tendency has been decreasing for the last 40–50 years in Europe. Eighteen studies specifically analyzing the development over the last 20 years are reviewed. The studies preponderantly show significant decreases most probably due to the reduction of the number of available psychiatric beds alongside the decentralization of psychiatry, and to decreasing virulence of schizophrenia. Alternatively, a diagnostic delay of schizophrenia can hardly explain the decrease.
Next day effects after nighttime treatment with zolpidem: a review
Authors: Undén M , Roth Schechter B
Published: July 2002

The purpose of this review was to analyze the literature for potential next-day residual effects of zolpidem, a non-benzodiazepine hypnotic, following nighttime administration. Based on more than 30 international clinical trials involving more than 2,600 subjects/patients, it can be concluded that at the recommended doses of zolpidem 10 mg for adults and zolpidem 5 mg for the elderly, at single or repeated dosing, in healthy subjects or insomniac patients, zolpidem appears to induce minimal next-day residual effects. As for all sedative hypnotics, zolpidem is indicated for the short-term treatment of insomnia and is recommended to be taken only when the patient is able to get a full night's sleep before resuming usual activities.
Redox‐dependent signal transduction
Authors: Finkel Toren
Published: July 2002

Reactive oxygen species (ROS) such as superoxide anions and hydrogen peroxide appear to be transiently produced in response to growth factor and cytokine stimulation. A variety of evidence suggests that this ligand‐stimulated change in the cellular redox state participates in downstream signal transduction. This review will focus on the effects of ROS on signal transduction pathways, the molecules that regulate intracellular ROS production and the potential protein targets of oxidants.
Cloning and characterization of human histone deacetylase 8
Authors: Van den Wyngaert Ilse , de Vries Winfred , Kremer Andreas , Neefs Jean-Marc , Verhasselt Peter , Luyten Walter H.M.L. , Kass Stefan U.
Published: July 2002

To date, seven different human histone deacetylases (HDACs) have been identified, which fall into two distinct classes. We have isolated and characterized a cDNA encoding a novel human HDAC, which we name HDAC8. HDAC8 shows a high degree of sequence similarity to HDAC1 and HDAC2 and thus belongs to the class I of HDACs. HDAC8 is expressed in a variety of tissues. Human cells overexpressing HDAC8 localize the protein in sub‐nuclear compartments whereas HDAC1 shows an even nuclear distribution. In addition, the HDAC8 gene is localized on the X chromosome at position q13, which is close to the XIST gene and chromosomal breakpoints associated with preleukemia.
Differential inhibition of IL‐6‐type cytokine‐induced STAT activation by PMA
Authors: Terstegen Lara , Maassen Björn G. , Radtke Simone , Behrmann Iris , Schaper Fred , Heinrich Peter C. , Graeve Lutz , Gatsios Petros
Published: July 2002

Prior activation of mitogen‐activated protein kinases by phorbol 13‐myristate 12‐acetate (PMA) results in an inhibition of interleukin (IL)‐6‐induced activation of the Janus kinase/signal transducer and activator of transcription (STAT) signaling pathway which is most likely mediated by the induction of suppressor of cytokine signaling‐3 and requires the specific SHP2 binding site Y759 of the IL‐6 signal transducer gp130. In this study, we demonstrate that PMA inhibits STAT activation by IL‐6 and the related cytokine leukemia inhibitory factor (LIF) but not by oncostatin M (OSM). Since the LIF receptor also contains an SHP2 recruitment site whereas the OSM receptor lacks such a module, we propose that two SHP2 binding modules within a homo‐ or heterodimeric receptor are necessary to mediate the PMA inhibitory effect.
Activation of p38 mitogen‐activated protein kinase is crucial in osteoclastogenesis induced by tumor necrosis factor
Authors: Matsumoto Masahito , Sudo Tatsuhiko , Maruyama Masumi , Osada Hiroyuki , Tsujimoto Masafumi
Published: July 2002

Tumor necrosis factor (TNF) induces osteoclast differentiation from bone marrow cells in the presence of macrophage colony‐stimulating factor. Treatment of bone marrow cells with SB203580 but not PD98059 inhibited TNF‐induced osteoclast differentiation. In RAW264 cells which differentiate into osteoclast‐like multinucleated cells by TNF treatment alone, activation of p38 mitogen‐activated protein (MAP) kinase induced by murine TNF was comparable to and independent of the receptor activator of necrosis factor‐κB ligand. Moreover, the number of multinucleated osteoclasts induced by TNF in bone marrow cell cultures derived from p38 MAP kinase gene deficient mice was significantly less than that from control mice. These results indicate that the p38 MAP kinase pathway plays a crucial role in TNF‐mediated osteoclast differentiation.
Differential regulation of apoptosis in AK‐5 tumor cells by the proto‐oncogene Bcl‐2: presence of Bcl‐2 dependent and independent pathways
Authors: Anjum Rana , Khar Ashok
Published: July 2002

The anti‐apoptotic protein Bcl‐2 functions as a crucial negative regulator of apoptosis. Bcl‐2 has been shown to prevent the efflux of apoptogenic factors from mitochondria to cytosol, thus inhibiting cell death. Here, we show the susceptibility of a spontaneously regressing, rat histiocytic tumor cell line, AK‐5, to the apoptotic effects of diverse stimuli and the ability of Bcl‐2 overexpression to block cell death. Bcl‐2 overexpression selectively inhibits apoptosis induced by ceramide and serum factor from AK‐5 tumor regressing animals but not actinomycin D and curcumin, whereas the pancaspase inhibitor z‐Val‐Ala‐Asp fluoromethylketone completely blocks apoptosis, irrespective of the inducer used. The ability of Bcl‐2 overexpression to block cell death does not depend on its ability to prevent cytochrome c release but correlates with its ability to prevent the dissipation of mitochondrial transmembrane potential. The results demonstrate that there are inducer dependent redundant activation pathways in a single cell, which may either be Bcl‐2 dependent or independent.
Inhibition of epithelial Na+currents by intracellular domains of the cystic fibrosis transmembrane conductance regulator
Authors: Kunzelmann K , Kiser G.L , Schreiber R , Riordan J.R
Published: July 2002

Cystic fibrosis is characterized by an impaired cyclic adenosine 3,5‐monophosphate (cAMP) activated Cl−conductance in parallel with an enhanced amiloride sensitive Na+conductance (ENaC) of the respiratory epithelium. Very recently, acute downregulation of ENaC by the cystic fibrosis transmembrane conductance regulator (CFTR) was demonstrated in several studies. The mechanism, however, by which CFTR exerts its inhibitory effect on ENaC remains obscure. We demonstrate that cytosolic domains of human CFTR are sufficient to induce inhibition of rat epithelial Na+currents (rENaC) when coexpressed inXenopusoocytes and stimulated with 3‐isobutyl‐1‐methylxanthine (IBMX). Moreover, mutations of CFTR, which occur in cystic fibrosis, abolish CFTR‐dependent downregulation of rENaC. Yeast two hybrid analysis of CFTR domains and rENaC subunits suggest direct interaction between the proteins. Enhanced Na+transport as found in the airways of cystic fibrosis patients is probably due to a lack of CFTR dependent downregulation of ENaC.
The crystal structure of human cathepsin L complexed with E‐64
Authors: Fujishima Akira , Imai Yumi , Nomura Toshiyuki , Fujisawa Yukio , Yamamoto Yoshio , Sugawara Tohru
Published: July 2002

We have determined the three dimensional structure of the complex of human cathepsin L and E‐64, an irreversible inhibitor of cysteine proteases, at 2.5 Å resolution. The overall structure was similar to that of other known cysteine proteases and apparently identical to the mature region of procathepsin L. The electron density for E‐64 is clearly visible except for the guanidinobutane moiety. From comparison of the active sites of cathepsin L and B, we found the following: (1) The S′ subsites of cathepsin L and B are totally different because of the `occluding loop' lying on the end of the S′ subsites of cathepsin B. (2) The S2 pocket of cathepsin L is shallow and narrow compared to that of cathepsin B. (3) The S3 subsites of the two enzymes are more similar than the other subsites, but cathepsin L may accommodate a more bulky group at this site. Knowledge of the active site structure of cathepsin L should be helpful for the structure‐based design of potent and specific inhibitors which are of therapeutic importance.
vesl, a gene encoding VASP/Ena family related protein, is upregulated during seizure, long‐term potentiation and synaptogenesis1
Authors: Kato Akihiko , Ozawa Fumiko , Saitoh Yoshito , Hirai Keiko , Inokuchi Kaoru
Published: July 2002

We have isolated a novel cDNA, vesl, that was induced during convulsive seizure in the rat hippocampus. The vesl gene encodes a protein of 186 amino acids that has significant homology to the EVH1 domain of the VASP/Ena family of proteins implicated in the control of microfilament dynamics. The expression of vesl mRNA was induced in the granule cell layer during persistent long‐term potentiation (LTP) of the dentate gyrus in an NMDA receptor‐dependent manner. Furthermore, vesl mRNA was expressed at a high level during hippocampal synaptogenesis. We suggest that the Vesl protein may be involved in the structural changes that occur at synapses during long‐lasting neuronal plasticity and development.
Expression and subcellular localization of mouse 20S proteasome activator complex PA28
Authors: Soza Andrea , Knuehl Christine , Groettrup Marcus , Henklein Peter , Tanaka Keiji , Kloetzel Peter-M.
Published: July 2002

We have cloned the mouse PA28 proteasome activator cDNAs. Northern blot demonstrates high PA28 mRNA levels in liver, kidney and lung. mRNA levels are low in thymus, spleen and brain. In contrast, PA28 protein levels vary little between these tissues. Immunocytological analysis and cell fractionation experiments demonstrate that both subunits are almost equally distributed between the cytoplasm and the nucleus. Interestingly, PA28α spares nucleoli, while PA28β is strongly enhanced in the nucleolus. This indicates for the first time that the PA28α and PA28β subunits may serve nuclear functions which may be different from and independent of each other.
A non‐peptide compound which can mimic the effect of thrombopoietin via c‐Mpl
Authors: Kimura Tatsuya , Kaburaki Hiroshi , Tsujino Tomomi , Ikeda Yoshitaka , Kato Hideo , Watanabe Yoshinari
Published: July 2002

Thrombopoietin (TPO) is a cytokine which plays a central role in megakaryopoiesis and platelet production by binding to its cell surface receptor, termed c‐Mpl. In the present study, two benzodiazepinones that compete with the binding of TPO to the extracellular region of c‐Mpl were identified, and one of them stimulated the proliferation of a human TPO‐dependent megakaryocytic cell line, UT‐7/TPO. It stimulated the activation of signal transducer and activator of transcription 5 in UT‐7/TPO cells. These results suggest that a non‐peptide compound can mimic the effect of TPO via c‐Mpl.
Nuclear sphingomyelin protects RNA from RNase action
Authors: Micheli Marta , Albi Elisabetta , Leray Claude , Magni Mariapia Viola
Published: July 2002

Chromatin phospholipidic fraction, as previously demonstrated, shows the same localization as RNA inside the nuclei. DNase and RNase treatment of nuclei removed almost totally the DNA, 63% of RNA and caused a 50% loss of phospholipids. The aim of the present investigation is to study the fraction of RNase undigested nuclear RNA and its relationship with the phospholipids still present in the nuclei. Isolated hepatocyte nuclei were treated with Triton X‐100 and digested with RNase and DNase. The undigested nuclear material contained proteins (98%) and a small amount of RNA (1.7%), DNA (0.4%) and phospholipids (0.18%). The analysis of phospholipids showed the presence of two components only, namely phosphatidylcholine and sphingomyelin. In the same complex, the activity of sphingomyelin synthase, phosphatidylcholine‐dependent phospholipase C and neutral sphingomyelinase has been detected. Treatment of isolated RNA with neutral sphingomyelinase modified the RNA in RNase sensitive RNA, thus suggesting that the SM may represent a bridge between two RNA strands possibly regulating transcription.
Superoxide radical production by sponges Sycon sp
Authors: Peskin Alexander V , Labas Yulii A , Tikhonov Alexander N
Published: July 2002

Using the catechol Tiron as an O−⋅
2 scavenger, we showed that sea sponges (Sycon sp.) produce superoxide radicals in sea water at a high rate without any stimuli added. The rate of O−⋅
2 outflow from sponges to their water surroundings reaches a value of 0.5 nmol/min per sponge at pH 6.5. The generation of O−⋅
2 was inhibited by Cu,Zn‐superoxide dismutase, and restored by the addition of KCN. We also confirmed the abiotic production of O−⋅
2 in sea water, detected earlier with a different method by Petasne and Zika [Nature 325 (1987) 516–518].
Human serum, cysteine and histidine inhibit the oxidation of low density lipoprotein less at acidic pH
Authors: Patterson Rebecca A , Leake David S
Published: July 2002

Low concentrations of serum or interstitial fluid have been shown to inhibit the oxidation of low density lipoprotein (LDL) catalysed by copper or iron, and may therefore protect against the development of atherosclerosis. As atherosclerotic lesions may have an acidic extracellular pH, we have investigated the effect of pH on the inhibition of LDL oxidation by serum and certain components of serum. Human serum (0.5%, v/v), lipoprotein‐deficient human serum at an equivalent concentration and the amino acids l‐cysteine (25 μM) and l‐histidine (25 μM), but not l‐alanine (25 μM), inhibited effectively the oxidation of LDL by copper at pH 7.4, as measured by the formation of conjugated dienes. The antioxidant protection was reduced considerably at pH 6.5, and was decreased further at pH 6.0. These observations may help to explain why LDL becomes oxidised locally in atherosclerotic lesions in the presence of the strong antioxidant protection offered by extracellular fluid.