bluuebunny/crossref_metadata_2025 huggingface dataset:
Divergence and convergence of TGF‐β/BMP signaling
Authors: Miyazono Kohei , Kusanagi Kiyoshi , Inoue Hirofumi
Published: August 2002

The transforming growth factor‐β (TGF‐β) superfamily includes more than 30 members which have a broad array of biological activities. TGF‐β superfamily ligands bind to type II and type I serine/threonine kinase receptors and transduce signals via Smad proteins. Receptor‐regulated Smads (R‐Smads) can be classified into two subclasses, i.e. those activated by activin and TGF‐β signaling pathways (AR‐Smads), and those activated by bone morphogenetic protein (BMP) pathways (BR‐Smads). The numbers of type II and type I receptors and Smad proteins are limited. Thus, signaling of the TGF‐β superfamily converges at the receptor and Smad levels. In the intracellular signaling pathways, Smads interact with various partner proteins and thereby exhibit a wide variety of biological activities. Moreover, signaling by Smads is modulated by various other signaling pathways allowing TGF‐β superfamily ligands to elicit diverse effects on target cells. Perturbations of the TGF‐β/BMP signaling pathways result in various clinical disorders including cancers, vascular diseases, and bone disorders. © 2001 Wiley‐Liss, Inc.
Endoglin: An accessory component of the TGF‐β‐binding receptor‐complex with diagnostic, prognostic, and bioimmunotherapeutic potential in human malignancies
Authors: Fonsatti Ester , Vecchio Luigi Del , Altomonte Maresa , Sigalotti Luca , Nicotra Maria Rita , Coral Sandra , Natali Pier Giorgio , Maio Michele
Published: August 2002

Endoglin (CD105) is a cell membrane glycoprotein over‐expressed on highly proliferating endothelial cells in culture, and on endothelial cells of angiogenetic blood vessels within benign and malignant tissues. CD105 binds several factors of the Transforming Growth Factor (TGF)‐β superfamily, and its over‐expression modulates cellular responses to TGF‐β1. The complex of experimental findings accumulated in the last few years strongly indicate that CD105 is a powerful marker of angiogenesis, and that it might play a critical role in the pathogenesis of vascular diseases and in tumor progression. In this paper, we will review the structural, biological and functional features of CD105, as well as its distribution within normal and neoplastic tissues, emphasizing its foreseeable role as a molecular target for new diagnostic and bioimmunotherapeutic approaches in human malignancies. © 2001 Wiley‐Liss, Inc.
On‐line monitoring of wastewater quality: a review
Authors: Bourgeois Wilfrid , Burgess Joanna E , Stuetz Richard M
Published: August 2002

Real‐time monitoring of wastewater quality remains an unresolved problem to the wastewater treatment industry. In order to comply with increasingly stringent environmental regulations, plant operators as well as instrument manufacturers have expressed the need for new standards and improved comparability and reliability of existing techniques. A review of currently available methods for monitoring global organic parameters (BOD, COD, TOC) is given. The study reviews both existing standard techniques and new innovative technologies with the focus on the sensors' potential for on‐line and real‐time monitoring and control. Current developments of biosensors, optical sensors and sensor arrays as well as virtual sensors for the monitoring of wastewater organic load are presented and the interests and limitations of these techniques with respect to their application to the wastewater monitoring are discussed.

© 2001 Society of Chemical Industry
Preparation and use of a mesoporous silicate material for the removal of tetramethyl ammonium hydroxide (TMAH) from aqueous solution
Authors: Kelleher Brian P , Doyle Aidan M , O'Dwyer Thomas F , Hodnett Benjamin K
Published: August 2002

A cubic mesoporous silicate (CMS) was prepared, characterised, and assessed as an adsorbent for tetramethyl ammonium hydroxide (TMAH) from aqueous solution. The adsorption process was studied as a function of temperature and time. Sorption closely followed the Langmuir model. The adsorption of TMAH on CMS was endothermic and kinetic studies suggest that the overall rate of adsorption was pseudo‐second‐order. Pore diffusion effects contribute to limiting the overall rate of adsorption while at lower initial TMAH concentrations, film diffusion becomes more important. Desorption studies were carried out using water and methanol. Methanol was the superior desorbing agent.

© 2001 Society of Chemical Industry
Nitric oxide: Relation to integrity, injury, and healing of the gastric mucosa
Authors: Calatayud Sara , Barrachina Dolores , Esplugues Juan V.
Published: August 2002

Nitric oxide (NO) plays a multifaceted role in mucosal integrity. The numerous functions of NO and the double‐edged role played by NO in most of them provide a great complexity to the NO action. The three enzymatic sources of NO, neuronal NO‐synthase (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS), have been characterised in the gastrointestinal tract. The protective properties of the NO derived from constitutive NO‐synthases (eNOS and nNOS) have already been well established. Less clear is the role assigned to iNOS. The simplistic initial view of low levels of NO synthesised by constitutive NOS being protective while exaggerated NO levels after iNOS induction leading irremediably to cytotoxicity is being questioned by new evidence. As initially reported for constitutive NOS, iNOS activity may be associated to reduced leukocyte‐endothelium interaction and platelet aggregation as well as protection of mucosal microcirculation. Moreover, iNOS activity may be important to resolve inflammation by increasing apoptosis in inflammatory cells. It is entirely possible that a low level of expression of iNOS will reflect a positive host‐defense response to challenge, but that exaggerated or uncontrolled expression of iNOS itself becomes detrimental. There is no doubt about the protective role of NO in physiological conditions. However, when the mucosa is threatened, the role of NO becomes multiple and the final effect will probably depend on the nature of the insult, the environment involved, and the interaction with other mediators. Microsc. Res. Tech. 53:325–335, 2001. © 2001 Wiley‐Liss, Inc.
NO message from muscle
Authors: Grozdanovic Zarko
Published: August 2002

The synthesis of the free radical gas nitric oxide (NO) is catalyzed by the enzyme NO synthase (NOS). NOS converts arginine and molecular oxygen to NO and citrulline in a reaction that requires NADPH, FAD, FMN, and tetrahydrobiopterin as cofactors. Three types of NOS have been identified by molecular cloning. The activity of the constitutively expressed neuronal NOS (nNOS) and endothelial NOS (eNOS) is Ca2+/calmodulin‐dependent, whereas that the inducible NOS (iNOS) is Ca2+‐insensitive. The predominant NOS isoform in skeletal muscle is nNOS. It is present at the sarcolemma of both extra‐ and intrafusal muscle fibers. An accentuated accumulation of nNOS is found in the endplate area. This strict sarcolemmal localization of nNOS is due its association with the dystrophin‐glycoprotein complex, which is mediated by the syntrophins. The activity of nNOS in skeletal muscle is regulated by developmental, myogenic, and neurogenic influences. NO exerts several distinct effects on various aspects of skeletal muscle function, such as excitation‐contraction coupling, mitochondrial energy production, glucose metabolism, and autoregulation of blood flow. Inside the striated muscle fibers, NO interacts directly with several classes of proteins, such as soluble guanylate cyclase, ryanodine receptor, sarcoplasmic reticulum Ca2+‐ATPase, glyceraldehyde‐3‐phosphate dehydrogenase, and mitochondrial respiratory chain complexes, as well as radical oxygen species. In addition, NO produced and released by contracting muscle fibers diffuses to nearby arterioles where it acts to inhibit reflex sympathetic vasoconstriction. Microsc. Res. Tech. 55:148–153, 2001. © 2001 Wiley‐Liss, Inc.
Temperature effects and genotype‐temperature interactions on sex determination in the European sea bass (Dicentrarchus labrax L.)
Authors: Saillant Eric , Fostier Alexis , Haffray Pierrick , Menu Bruno , Thimonier Jacques , Chatain Béatrice
Published: August 2002

The effect of temperature on sex‐ratios in 27 families of sea bass reared in the same tank from the fertilization stage onward was investigated. An excess of males (68%) was found in the groups that were reared at high temperature (mean�±�standard deviation: 20�±�1°C) until they reached the mean size of 8.1 cm (Standard Length, 149 days post‐fertilization [p.f.]). Masculinization was higher (89% of males) in the groups maintained at low temperature (13°C), from fertilization to a mean length of 6.5�cm (346 days p.f.). Shifts from high to low temperature at 8.1�cm and from low to high temperature at 6.5�cm had no consequence on the sex‐ratio. The percentage of males showing intratesticular oocytes was higher at low temperature (63%) than at high temperature (36%), suggesting that these males may be sensitive fish that have been masculinized by environmental factors. Fish sampled in the groups reared at high (2,200 fish) and low (500 fish) temperature were genotyped on three microsatellite loci. This allowed them to be assigned to the breeders used in the crossing design, thus permitting an analysis of parental influence on sex‐ratios. In groups reared at high temperature, both parents had a significant additive effect on the percentage of females, and the interaction between sire and dam was not significant. Genotype temperature interactions were also detected and their existence suggests the interesting possibility of selecting nonsensitive genotypes in breeding programs. J. Exp. Zool. 292:494–505, 2002. © 2002 Wiley‐Liss, Inc.
Temperature sex determination in the European sea bass, Dicentrarchus labrax (L., 1758) (Teleostei, Perciformes, Moronidae): Critical sensitive ontogenetic phase
Authors: Koumoundouros George , Pavlidis Michalis , Anezaki Lina , Kokkari Constantina , Sterioti Aspasia , Divanach Pascal , Kentouri Maroudio
Published: August 2002

The temperature sex determination (TSD) mechanism in the European sea bass (Dicentrarchus labrax L.) was studied in respect to: a) the TSD sensitivity during the different developmental stages; and b) the intrapopulation correlation of sex determination with the growth rate up to the end of the TSD‐sensitive period. At the stage of half‐epiboly, eggs from the same batch were divided into four groups and subjected to different thermal treatments: a) 15°C (G15 group) and b) 20°C (G20 group) up to the middle of metamorphosis stage; c) 15°C up to the end of yolk‐sac larval stage and subsequently to 20°C (G15‐5 group); and d) 15°C up to the end of the preflexion stage and then to 20°C (G15‐10 group). At the end of the treatments, size grading was applied and four additional populations were established from the upper (L) and lower (S) size portions of the G15 and G20 populations: G15L, G15S, G20L, and G20S. During the following growing phase, all populations were subjected to common rearing conditions. The sex ratios of each population were macroscopically determined at 190–210 mm mean total length. Female incidence was significantly affected (P < 0.05) by the different thermal treatments: 66.1% in the G15, 47.1% in the G15‐10, 37.6% in the G15‐5, and 18.1% in the G20 group. In addition, sex ratio was correlated with the growth rate of the fish up to the end of the TSD‐sensitive period, with the larger fish presenting a significantly higher (P < 0.01) female incidence than the smaller fish in both thermal regimes tested: 73.1% in G15L vs. 57% in G15S, and 36.6% in G20L vs. 22.5% in G20S group. Results provide, for the first time, clear evidence that the sea bass is sensitive to TSD during all different ontogenetic stages up to metamorphosis, and that sex ratio is correlated with the growth rate of the fish well before the differentiation and maturation of the gonads. J. Exp. Zool. 292:573–579, 2002. © 2002 Wiley‐Liss, Inc.
Cell signaling and ion transport across the fish gill epithelium
Authors: Evans David H.
Published: August 2002

A large array of circulating and local signaling agents modulate transport of ions across the gill epithelium of fishes by either affecting transport directly or by altering the size and distribution of transporting cells in the epithelium. In some cases, these transport effects are in addition to cardiovascular effects of the same agents, which may affect the perfusion pathways in the gill vasculature and, in turn, affect epithelial transport indirectly. Prolactin is generally considered to function in freshwater, because it is the only agent that allows survival of some hypophysectomized fish species in freshwater. It appears to function by either reducing branchial permeability, Na,K‐activated ATPase activity, or reducing the density of chloride cells. Cortisol was initially considered to produce virtually opposite effects (e.g., stimulation of Na,K‐activated ATPase and of chloride cell size and density), but more recent studies have found that this steroid stimulates ionic uptake in freshwater fishes, as well as the activity of H‐ATPase, an enzyme thought to be central to ionic uptake. Thus, cortisol may function in both high and low salinities. Growth hormone and insulin‐like growth factor appear to act synergistically to affect ion regulation in seawater fishes, stimulating both Na,K‐activated ATPase and Na‐K‐2Cl co‐transporter activity, and chloride cell size, independent of their effects on growth. Some of the effects of the GH–IGF axis may be via stimulation of the number of cortisol receptors. Thyroid hormones appear to affect seawater ion regulation indirectly, by stimulating the GH–IGF axis. Natriuretic peptides were initially thought to stimulate gill ionic extrusion, but recent studies have not corroborated this finding, so it appears that the major mode of action of these peptides may be reduction of salt loading by inhibition of oral ingestion and intestinal ionic uptake. Receptors for both arginine vasotocin and angiotensin have been described in the gill epithelium, but their respective roles and importance in fish ion regulation remains unknown. The gill epithelium may be affected by both circulating and local adrenergic agents, and a variety of studies have demonstrated that stimulation of α‐adrenergic versus β‐adrenergic receptors produces inhibition or stimulation of active salt extrusion, respectively. Local effectors, such as prostaglandins, nitric oxide, and endothelin, may affect active salt extrusion as well as gill perfusion. Recent studies have suggested that the endothelin inhibition of salt extrusion is actually mediated by the release of both NO and prostaglandins. It is hoped that modern molecular techniques, combined with physiological measurements, will allow the dissection of the relative roles in ion transport across the fish gill epithelium of this surprisingly large array of putative signaling agents. © 2002 Wiley‐Liss, Inc.
Alterations in habituation of the tail flip response in epigean and troglobitic crayfish
Authors: Kellie Scott , Greer Jarrett , Cooper Robin L.
Published: August 2002

We demonstrate that the probability of the crayfish, P. clarkii, to tail flip in response to a touch on the dorsal tail fan is dependent on both the size and the behavioral state of the animal. Alterations in the animal’s internal physical state, such as when the animal autotomizes its chelipeds, will cause larger‐sized animals to tail flip; if they were not autotomized, then no tail flip response would occur. Altering the external environment by removal of water causes small crayfish, which normally habituate slowly, to rapidly habituate. Observation of large adult crayfish in a species, O. australis packardi, one that evolved to live in total cave darkness, revealed that they are more likely to tail flip than are the sighted, adult P. clarkii. Results indicate that the behavioral state of the crayfish can result in rapid and long‐term alterations in the tail flip response and in habituation rates to repetitive stimuli. This ability to show plasticity in gain setting may be regulated by neuromodulators and can occur in large adults of the sighted crayfish. Differences between the two species indicate that size may not be the sole contributing factor to account for tail flip behaviors. J. Exp. Zool. 290:163–176, 2001. © 2001 Wiley‐Liss, Inc.
Bafilomycin A1 inhibits proteolytic cleavage and hydration but not yolk crystal disassembly or meiosis during maturation of sea bass oocytes
Authors: Selman Kelly , Wallace Robin A. , Cerdà Joan
Published: August 2002

Oocytes of the black sea bass, Centropristes striata, were enlarged in volume more than three‐fold over a 24‐hr period during oocyte maturation, both in vivo and in vitro. At the same time, the opaque oocytes clarified while the crystalline yolk inclusions lost their ordered structure, fused with one another, and formed a continuous electron‐lucent mass. The oocyte size increase was due almost entirely to water uptake, which was accompanied by the accumulation of Na+, K+, and free amino acids (FAAs). The absolute amounts of each of these small molecular weight osmotic effectors increased 2×, 4×, and over 10×, respectively, indicating that the generation of FAAs is the major cause of water uptake during maturation. Amino acid analyses indicated that the amounts of all amino acids except taurine increased, so that selective amino acids were not produced during maturation. The increase in FAAs was accompanied by the loss of certain high‐molecular‐weight yolk proteins and the generation of many smaller peptides. Oocytes stimulated to undergo maturation in the presence of bafilomycin A1, a specific inhibitor of the vacuolar ATPase‐dependent proton pump, clarified and underwent maturation but did not increase significantly in size. Cytological examination revealed that yolk crystals fused and became homogeneous but maintained their electron density. No evidence of proteolysis was found in bafilomycin A1‐treated oocytes and the generation of FAAs together with hydration was inhibited in a dose‐dependent manner (I50 = 3 nM bafilomycin A1). Taken together, we postulate that the pronounced oocyte hydration in marine teleosts that spawn pelagic (floating) eggs is accomplished by a two‐step process whereby (i) K+ influx promotes yolk crystal disassembly and yolk sphere fusion and (ii) acidification of the yolk spheres activates yolk proteolysis and concomitant hydration. Bafilomycin A1 inhibits only the second step so that many of the events of oocyte maturation, including germinal vesicle breakdown, occur in its presence but oocyte hydration is suppressed. J. Exp. Zool. 290:265–278, 2001. © 2001 Wiley‐Liss, Inc.
Synthesis of 14C‐labelled sulforaphane
Authors: Kuhnert Nikolai , Holst Birgit , Williamson Gary
Published: August 2002

Sulforaphane (SFN) a naturally occurring isothiocyanate present in broccoli, shows strong evidence of anti‐carcinogenic activity. The mechanism of action, absorption, distribution, metabolism and excretion of the compound is however still poorly understood and requires a radiolabelled version of the compound for further studies. The paper describes an optimised cold synthesis of SFN and a radiosynthesis of 14C‐labelled SFN. Copyright © 2001 John Wiley & Sons, Ltd.
Synthesis of 10,11,14,15‐13C4‐and 14,15‐13C2‐retinyl acetate
Authors: Tanumihardjo Sherry A.
Published: August 2002

The synthesis of 10,11,14,15‐13C4‐ and 14,15‐13C2‐retinyl acetate is described. The procedure uses a modified Wittig‐Horner synthesis. By using the 13C isotope, sensitive gas chromatography‐combustion‐isotope ratio mass spectrometry can be used for analysis in application to human isotope dilution studies. Copyright © 2001 John Wiley & Sons, Ltd.
Synthesis of asymmetric [75Se]selenoethers via carbodiimides
Authors: Blum T. , Ermert J. , Coenen H. H.
Published: August 2002

A convenient radiosynthesis of asymmetric [75Se]selenoethers was developed using 1,3‐disubstituted [75Se]selenoureas as intermediates. These were prepared from appropriate carbodiimides and hydrogen [75Se]selenide, which could only be generated from carrier‐added (c.a.) [75Se]selenite in aqueous solution using phosphinic acid as reducing agent. Optimization of this initial labelling step with dicyclohexylcarbodiimide and polymeric N‐cyclohexylcarbodiimide‐N′‐methyl polystyrene resulted in radiochemical yields (RCY) of 73 and 55% (bound on the polymer), respectively, within 45 min. Treatment of [75Se]selenoureas with alkylbromides led to corresponding [75Se]selenouronium salts in nearly quantitative yields. Hydrolysis under basic conditions provided the [75Se]selenolates and a second alkylation yielded asymmetric [75Se]selenoethers. Thus, within 90 min benzylmethyl[75Se]selenide, benzylbutyl[75Se]selenide, benzylisopropyl[75Se]selenide and 1‐phenyl‐l‐(methyl[75Se]seleno)ethane were synthesized with respective RCY of about 59, 55, 10 and 60%. Furthermore, the 75Se‐labelled alkylating agent 3‐(methyl[75Se]seleno)‐1‐propanyl p‐toluenesulfonate and [75Se]selenomethionine were obtained with radiochemical yields of 51 and 41%, respectively. Copyright © 2001 John Wiley & Sons, Ltd.
Testis differentiation in the glowworm, Lampyris noctiluca, with special reference to the apical tissue
Authors: Balles S. , Maas U. , Sehn E. , Dorn A.
Published: August 2002

The gonads of Lampyris noctiluca are sexually undifferentiated during the first larval instars. They consist of many gonadal follicles that include the germ stem cells enclosed by the somatic cells of the follicle wall. Follicle wall cells are more numerous at the follicle apices than at the distal parts, but different cell types cannot be distinguished. In male larvae, the appearance of apical follicle tissue, derived from follicle wall cells, marks the onset of testis differentiation. When maximally expressed, the apical tissue occupies about the upper half of the testis follicles and can be observed in larvae of the fifth and sixth instar. The apical tissue is characterized by its “light” appearance (due to poor stainability) caused by the small number cellular organelles, especially a paucity of free ribosomes. Maximal expression of the apical tissue must be very brief, since in most examined fifth and sixth instar larvae the apical tissue is partly or mostly translocated into the center of the upper half of the follicles and spermatogonia then occupy the apical follicle tips. During and after translocation apical cells form projections that grow around clusters of spermatogonia (spermatocysts). Thus, the apical cells transform into spermatocyst envelope cells. They retain their “light” appearance but undergo dramatic subcellular differentiation: smooth ER becomes extremely prominent, forming stacks and whorls of parallel cisternae. Golgi complexes are also conspicuous. The cellular organization suggests secretory activity. The possibility of ecdysteroid production and its function is discussed. The spermatocyst envelope cells persist into the pupal stage. When spermiohistogenesis takes place in cysts, cyst envelope cells show signs of regression. At all stages of testis development apical cells and their derivatives, the spermatocyst envelope cells, phagocytize degenerating spermatogonia. Although this is an important task of these cells, the impressive formation of sER in the cyst envelope cells is indicative of an additional, as yet unknown, function. J. Morphol. 251:22–37, 2002. © 2002 Wiley‐Liss, Inc.
Relaxation effects of clustered particles
Authors: Tanimoto Akihiro , Oshio Koichi , Suematsu Makoto , Pouliquen Daniel , Stark David D.
Published: August 2002

Relations between spatial distribution of superparamagnetic iron oxide (SPIO) particles and the image contrast caused by SPIO were investigated. Actual clustering pattern of particles was measured in the liver and spleen of animals using intravital laser confocal microscopy. SPIO‐doped phantoms with and without Sephadex beads were made to simulate these patterns, and relaxation parameters were measured using a 1.5‐T clinical scanner. Finally, these results were compared to clinical image data using SPIO particulate agent. Intravital microscopy indicated that the clustering of latex beads was more predominant in hepatic Kupffer cells than in splenic macrophages (P < 0.001). Phantoms without Sephadex beads showed an approximately linear increase of 1/T1 (R1), 1/T2 (R2) and 1/T2* (R2*) values with increasing SPIO concentration. However, with Sephadex beads, R1 and R2 showed little change with increasing SPIO concentration, while R2* showed the same linear increase with SPIO. Also, the R2* values were higher with Sephadex beads. These results were consistent with the clinical imaging data, where signal reduction was significantly smaller in the spleen (‐0.4% ± 27.4%) than in the liver (50.4% ± 16.8%, P < 0.00001) on T2*‐weighted images, but the reduction in the spleen (47.2% ± 16.1%) was equivalent to the liver (38.8% ± 26.0%) on T2‐weighted images. J. Magn. Reson. Imaging 2001;14:72–77. © 2001 Wiley‐Liss, Inc.
Sequencing of new beauverolides by high‐performance liquid chromatography and mass spectrometry
Authors: Kuzma Marek , Jegorov Alexandr , Kačer Petr , Havlíček Vladimír
Published: August 2002

Mass spectrometry (MS) and tandem mass spectrometry (MSn) were used for the identification of beauverolides in the fermentation broth of Beauveria bassiana and for evaluation of the purified fraction obtained by sublimation of beauverolides. Besides being a new efficient route for purification of beauverolides, sublimation provided an enrichment of new minor lipophilic beauverolides of lower molecular weight from the original complex mycelial extract. The product ion collision‐induced dissociation (CID) spectra obtained on an ion trap (electrospray ionization), the in‐source CID mass spectra on a sector instrument (atmospheric‐pressure chemical ionization) and the post‐source decay matrix‐assisted laser desorption/ionization mass spectra of beauverolides were compared and evaluated. All MSn experiments started with singly charged precursor ions. The following two new representatives of this group of compounds were identified by high‐performance liquid chromatography and MS (HPLC/MS): cyclo‐(3‐hydroxy‐4‐methyloctanoylvalylalanylleucyl) and cyclo‐(3‐hydroxy‐4‐methyloctanoyltyrosylalanylleucyl). Individual structures were confirmed by preparative isolation and nuclear magnetic resonance spectroscopy. The structure of a third novel and minor beauverolide was tentatively assigned by HPLC/MS only as cyclo‐(3‐hydroxy‐4‐methyldecanoylvalylalanylLxx), Lxx = leucyl, isoleucyl, or allo‐isoleucyl. Copyright © 2001 John Wiley & Sons, Ltd.
Characterization of a phosphorylated peptide and peptoid and peptoid–peptide hybrids by mass spectrometry
Authors: Ruijtenbeek Rob , Versluis Cees , Heck Albert J. R. , Redegeld Frank A. M. , Nijkamp Frans P. , Liskamp Rob M. J.
Published: August 2002

Nano‐electrospray tandem mass spectrometry (nano‐ES‐MS/MS) was used to record collision‐induced dissociation (CID) spectra of a set of peptoid–peptide hybrids and the complete peptoid derived from the phosphopeptide Ac‐pTyr‐Glu‐Thr‐Leu‐NH2 (1). The presence of B and Y″‐type fragment ions in the tandem mass spectra of the protonated molecular ions [M + H]+ allowed confirmation of sequence similar to mass spectrometric sequence analysis in peptides. In the isomeric peptoid compounds studied, one or several amino acid residues were replaced by peptoid residues (N‐substituted glycine residues), which resulted in characteristic tandem mass spectra with differently increased relative abundances of Y″‐and B‐type fragment ions. The increment of a particular Y″‐ion was directly correlated to the position of a peptoid residue present. In addition to these increased peak intensities, other characteristic peaks were also observed compared with the spectrum of reference peptide 1. When a peptoid phosphotyrosine was incorporated, the presence of this residue was apparent from the occurrence of a relatively intense peak at m/z 187 representing the positively charged side‐chain of phosphotyrosine, which was almost absent in the spectrum of the reference peptide 1. Since the threonine side‐chain had to be translated into the homo peptoid analog this substitution was apparent from the presence of [M + H]+ and fragment ions 14 mass units higher than observed in the spectrum of the reference phosphopeptide 1. The presence of an NLeu peptoid residue could be confirmed by the specific fragmentation of the immonium ion showing an intense peak in its tandem mass spectrum at m/z 57, which results from the loss of an neutral imine molecule leading to a positively charged [C4H9]+ ion. By means of these mass spectrometric characteristics, all isomeric peptoid compounds could be distinguished from each other and characterized. The methods used appear to be very useful in future studies of peptoids and peptoid–peptide hybrids. Copyright © 2001 John Wiley & Sons, Ltd.
Characteristics of puumala and Dobrava infections in Croatia*
Authors: Markotić A. , Nichol S.T. , Kuzman I. , Sanchez A.J. , Ksiazek T.G. , Gagro A. , Rabatić S. , Zgorelec R. , Avšič‐Županc T. , Beus I. , Dekaris D.
Published: August 2002

In this study, two different hantaviruses, Puumala virus (PUUV) and Dobrava virus (DOBV), were demonstrated for the first time to coexist and cause hemorrhagic fever with renal syndrome (HFRS) in Croatia. Phylogenetic analysis showed some differences among the nucleotide sequences of PUUV originating from Dinara mountain, which was more closely related to Austrian PUUV than other Croatian PUUV from Mala Kapela mountain. More consistency was found among the Croatian DOBV. HFRS was verified in 85 of 201 suspected cases recorded in 1995 during the largest HFRS outbreak in Croatia. Most of these cases were soldiers. With the exception of the coastal region and islands, all of Croatia was found to be an area endemic for HFRS. A statistically significantly higher proportion of DOBV‐infected patients had acute renal failure, visual disturbance, severe thrombocytopenia, and elevated levels of nonsegmented leukocytes, creatine, and total bilirubin. The prevalence of gastrointestinal and electrocardiography disorders also was greater in DOBV‐infected patients. Interestingly, significantly more PUUV‐infected patients had elevated systolic blood pressure on admission to the hospital. Further prospective studies are necessary to shed more light on differences in HFRS severity associated with PUU and DOB viruses. J. Med. Virol. 66:542–551, 2002. © 2002 Wiley‐Liss, Inc.
Cytokine gene expression and protein production in peripheral blood mononuclear cells of children with acute dengue virus infections
Authors: Gagnon Susan J. , Mori Masuko , Kurane Ichiro , Green Sharone , Vaughn David W. , Kalayanarooj Siripen , Suntayakorn Saroj , Ennis Francis A. , Rothman Alan L.
Published: August 2002

Plasma leakage in dengue hemorrhagic fever (DHF) is associated with elevated plasma levels of cytokines. To define further the contribution of immune activation to DHF and the source of cytokines, we analyzed the production of cytokines in peripheral blood mononuclear cells (PBMC) obtained from children with dengue, using RT‐PCR and immunostaining. Tumor necrosis factor‐α (TNF‐α) and TNF‐β expression was detected in all samples by PCR and in < 50% of samples by immunostaining. Interferon‐γ (IFN‐γ) expression was detected in < 50% of samples by either method. Interleukin‐2 (IL‐2) and IL‐4 expression was detected in a few samples by immunostaining but was not detectable by PCR. We found greater expression of TNF‐α and IL‐4 in DHF than in dengue fever or other (non‐dengue) febrile illnesses. These results support the model of immunopathogenesis of DHF. However, low levels of cytokine expression in PBMC suggest that cellular activation in tissues may contribute to high serum cytokine levels in DHF. J. Med. Virol. 67:41–46, 2002. © 2002 Wiley‐Liss, Inc.
Novel variants related to TT virus distributed widely in China*
Authors: Luo Kangxian , He Haitang , Liu Zhihua , Liu Dingxie , Xiao Hong , Jiang Xiaojing , Liang Weifang , Zhang Lian
Published: August 2002

TTV is a DNA virus with high genetic heterogeneity. To investigate the novel isolates of the virus, blood samples were collected from subjects who lived in various parts of China and suffered from hepatitis or were asymptomatic carriers. Nested PCR was carried out to amplify a 3.2‐kb fragment using primers deduced from the prototype TTV (TA278). The ten entire 3.2‐kb nt sequences were aligned with isolate TA278, SANBAN, TUS01, and SENV retrieved from GenBank, and a phylogenetic tree was constructed by Neighbor‐Joining method. The analysis indicated that five novel variants of the present study have not been described before, and all TTV‐related isolates could be classified into three groups. The isolate TCHN‐A, B and TUS01 were included in a group, and the remaining novel isolates together with SANBAN and TA278 clustered into another group, while SEN virus formed a distinct group. The genetic distances of the five novel variants were 0.5507–0.8476 to TA278, 0.4635–0.7877 to SANBAN, 0.6064–0.7834 to TUS01 and 0.6936–0.8236 to SENV. Of these novel variants, the ORF1 consisted of 426–772 aa and ORF2 of 141–156 aa. The nt identities of ORF1 and ORF2 between those variants and TA278, SANBAN, and TUS01 were 46.1–60.8 and 48.7–63.6%, and those of aa sequences were only 27.1–52.4 and 28.9–45.5%, respectively. The first 65 aa of ORF1 were rich in arginine and most conserved with homology of 56.5–70.0%. There was a hypervariable region from aa 286 to 403 with merely 17.7–27.0% of identity. Despite a low aa identity between TA278 and the variants, they have similar hydrophilicity profiles of ORF1. There were 2–10 N‐glycosylation motifs found in these variants. In conclusion, despite the high divergence, sequences of all these isolates shared common genome organisation, ORF structure, hydrophilicity patterns, and some potential motifs with TTV prototype. It is suggested that various TTV and TTV‐related isolates belong to a very large and complex family, which remains to be studied. J. Med. Virol. 67:118–126, 2002. © 2002 Wiley‐Liss, Inc.
Differential effects of the nonsteroidal antiinflammatory drug flurbiprofen and its nitric oxide‐releasing derivative, nitroflurbiprofen, on prostaglandin E2, interleukin‐1β, and nitric oxide synthesis by activated microglia
Authors: Ajmone‐Cat Maria Antonietta , Nicolini Alessia , Minghetti Luisa
Published: August 2002

Increasing experimental, clinical, and epidemiological studies point to the pivotal role of inflammation in the pathogenesis of acute and chronic neurodegenerative diseases and to the protective effects of nonsteroidal antiinflammatory drug (NSAID) therapies. Nonetheless, NSAID long‐term therapies are limited by their significant adverse effects on gastrointestinal tract and kidneys. Nitroflurbiprofen (NO‐flurbiprofen) belongs to a novel class of antiinflammatory agents obtained by derivatization of conventional NSAIDs with a nitric oxide (NO)‐releasing moiety, which strongly reduces their untoward side effects without altering the antiinflammatory effectiveness. The recent evidence of neuroprotective effects of NO‐NSAIDs in animal models of chronic brain inflammation prompted us to investigate the activities of NO‐flurbiprofen and its parent molecule flurbiprofen on activated rat microglia, the brain resident macrophages. We found that NO‐flurbiprofen was as potent as flurbiprofen in preventing prostaglandin E2 synthesis in lipopolysaccharide‐activated microglial cultures. At variance with previous observations on peripheral macrophages is that NO‐flurbiprofen did not show any additional capacity to inhibit interleukin‐1β synthesis compared with flurbiprofen. Moreover, NO enhanced the expression of the inducible NO synthase; this effect was most likely attributable to the NO released from the drug, as suggested by experiments performed in the presence of the NO donor Deta‐NONOate, which similarly to NO‐flurbiprofen is characterised by a slow and long‐lasting release. Our findings indicate that NO‐NSAIDs may differently affect peripheral and brain macrophages. Given their potential therapeutic role in brain inflammation, further in vivo and in vitro studies are required to understand fully their mechanism of action in the CNS. J. Neurosci. Res. 66:715–722, 2001. © 2001 Wiley‐Liss, Inc.
Mitochondrial impairment in the cerebellum of the patients with progressive supranuclear palsy
Authors: Park Larry C.H. , Albers David S. , Xu Hui , Lindsay J. Gordon , Beal M. Flint , Gibson Gary E.
Published: August 2002

Abnormalities in energy metabolism and oxidative stress accompany many neurodegenerative diseases, including progressive supranuclear palsy (PSP). Previously, we showed decreased activities of a mitochondrial enzyme complex, α‐ketoglutarate dehydrogenase complex (KGDHC), and marked increases in tissue malondialdehyde levels in post‐mortem superior frontal cortex from the patients with PSP. The current study demonstrates that KGDHC is also significantly diminished (−58%) in the cerebellum from patients with PSP (n = 14), compared to age‐matched control brains (n = 13). In contrast to cortex, markers of oxidative stress, such as malondialdehyde, tyrosine nitration or general protein carbonyl modification, did not increase in cerebellum. Furthermore, the protein levels of the individual components of KGDHC did not decline. The activities of two other mitochondrial enzymes were measured to determine whether the changes in KGDHC were selective. The activity of aconitase, a mitochondrial enzyme with an iron/sulfur cluster, is also significantly diminished (−50%), whereas glutamate dehydrogenase activity is unchanged. The present results suggest that the interaction of metabolic impairment and oxidative stress is region‐specific in PSP brain. In cerebellum, reductions in KGDHC occur in the absence of increases in common measures of oxidative stress, and may underlie the metabolic deficits and contribute to pathological and clinical manifestation related to the cerebellum in patients with PSP. © 2001 Wiley‐Liss, Inc.
Phosphatidylinositol kinase enzymes regulate the retrograde axonal transport of NT‐3 and NT‐4 in sympathetic and sensory neurons
Authors: Bartlett Selena E. , Reynolds Anna J. , Weible Michael , Hendry Ian A.
Published: August 2002

Phosphatidylinositol 3‐kinase (PI3‐kinase) and phosphatidylinositol 4‐kinase (PI4‐kinase) enzymes are an important family of signaling molecules that have been implicated in the regulation of intracellular vesicle trafficking. It has previously been shown that PI3‐kinase and PI4‐kinase enzymes regulate neuronal survival and the retrograde axonal transport of nerve growth factor in sympathetic and sensory neurons. We have extended these studies to examine the role these enzymes play in the regulation of the retrograde axonal transport of neurotrophin‐3 (NT‐3) and neurotrophin‐4 (NT‐4) in sympathetic and sensory neurons in vivo. Wortmannin (0.1 nmol/eye), a PI3‐kinase and PI4‐kinase antagonist, reduced the amount of 125I‐NT‐3 retrograde transport in sympathetic neurons by approximately 50% and 125I‐NT‐4 in sympathetic neurons by approximately 40% and sensory neurons by approximately 20%. The PI3‐kinase antagonist LY294002 (100 nmol/eye) reduced the retrograde axonal transport of 125I‐NT‐4 in sympathetic and sensory neurons, and 125I‐NT‐3 in sympathetic neurons. Phenylarsine oxide (PAO), a PI4‐kinase antagonist, significantly inhibited 125I‐NT‐4 retrograde axonal transport in sympathetic and sensory neurons. These results show that wortmannin‐sensitive PI3‐kinases and PI4‐kinases may be involved in NT‐3 and NT‐4 retrograde axonal transport. The retrograde axonal transport of neurotrophic factors in sympathetic and sensory neurons in vivo appears to depend upon the activation of different receptors and second messenger cascades at the nerve terminal. © 2002 Wiley‐Liss, Inc.
Nitric oxide and nitric oxide synthase in Huntington's disease
Authors: Deckel A. Wallace
Published: August 2002

Nitric oxide (NO) is a biologically active inorganic molecule produced when the semiessential amino acid l‐arginine is converted to l‐citrulline and NO via the enzyme nitric oxide synthase (NOS). NO is known to be involved in the regulation of many physiological processes, such as control of blood flow, platelet adhesion, endocrine function, neurotransmission, neuromodulation, and inflammation, to name only a few. During neuropathological conditions, the production of NO can be either protective or toxic, dependent on the stage of the disease, the isoforms of NOS involved, and the initial pathological event. This paper reviews the properties of NO and NOS and the pathophysiology of Huntington's disease (HD). It discusses ways in which NO and NOS may interact with the protein product of HD and reviews data implicating NOS in the neuropathology of HD. This is followed by a synthesis of current information regarding how NO/NOS may contribute to HD‐related pathology and identification of areas for potential future research. J. Neurosci. Res. 64:99–107, 2001. © 2001 Wiley‐Liss, Inc.
Expression of estrogen receptor (ER)‐α and ‐β immunoreactivity in hippocampal cell cultures with special attention to GABAergic neurons
Authors: Su Jun‐Da , Qiu Jian , Zhong Yong‐Ping , Li Xiao‐Yue , Wang Jian‐Wen , Chen Yi‐Zhang
Published: August 2002

This study investigated the expression patterns of estrogen receptor‐α (ERα) and ‐β (ERβ) in the cultured hippocampal cells of neonatal rats by combined application of cell culture and immunocytochemistry. The results revealed that the expression difference between ERα and ERβ seemed to be not obvious in the cultured hippocampal cells of neonatal rats. Moreover, immunoreactivity for either ERα or ERβ was observed to be localized in the majority of not only neurons but also astrocytes. The coexpression of both ERα and ERβ in the same individual cell was also demonstrated by the double‐label immunocytochemistry. Western blot analysis showed that immunoreactivity for ERα in the neonatal hippocampal tissues was much higher than in the adult (became rather weak), although there was not such a great difference of immunoreactivity for ERβ. The data also provide direct evidence for the expression of ER subtypes within GABAergic neurons in hippocampal cell cultures and suggest that estrogen's effect on the hippocampus may be mediated at least in part by its ER‐containing GABAergic neurons. J. Neurosci. Res. 65:396–402, 2001. © 2001 Wiley‐Liss, Inc.
Vitamin E suppression of microglial activation is neuroprotective
Authors: Li Yuekui , Liu Ling , Barger Steven W. , Mrak Robert E. , Griffin W. Sue T.
Published: August 2002

Neurotoxic microglial–neuronal interactions have been implicated in the pathogenesis of various neurodegenerative diseases such as Alzheimer's disease, and vitamin E has been shown to have direct neuroprotective effects. To determine whether vitamin E also has indirect neuroprotective effects through suppression of microglial activation, we used a microglial–neuronal coculture. Lipopolysaccharide (LPS) treatment of a microglial cell line (N9) induced a time‐dependent activation of both p38 mitogen‐activated protein kinase (p38 MAPK) and nuclear factor‐κB (NFκB), with consequent increases in interleukin‐1α (IL‐1α), tumor necrosis factor‐α (TNF‐α), and nitric oxide (NO) production. Differentiated neuronal cells (PC12 cells treated with nerve growth factor) exhibited marked loss of processes and decreased survival when cocultured with LPS‐activated microglia. Preincubation of microglia with vitamin E diminished this neurotoxic effect, independently of direct effects of the antioxidant on the neuronal cells. Microglial NO production and the induction of IL‐1α and TNFα expression also were attenuated by vitamin E. Such antiinflammatory effects of vitamin E were correlated with suppression of p38 MAPK and NFκB activation and were mimicked by an inhibition of either p38 MAPK (by SB203580) or NFκB (by decoy oligonucleotides). These results suggest that, in addition to the beneficial effects of providing direct antioxidant protection to neurons reported by others, vitamin E may provide neuroprotection in vivo through suppression of signaling events necessary for microglial activation. J. Neurosci. Res. 66:163–170, 2001. Published 2001 Wiley‐Liss, Inc.
Regional temperature variability in the European Alps: 1760–1998 from homogenized instrumental time series
Authors: Böhm Reinhard , Auer Ingeborg , Brunetti Michele , Maugeri Maurizio , Nanni Teresa , Schöner Wolfgang
Published: August 2002

This paper investigates temperature variability in the Alps and their surroundings based on 97 instrumental series of monthly mean temperatures. A discussion of the initial homogenizing procedure illustrates its advantages and risks. A comparison of the homogenized series with the original series clearly shows the necessity to homogenize. Each of the original series had breaks (an average of five per series) and the mean of all series was systematically biased by non‐climatic noise. This noise has subdued the long‐term amplitude of the temperature evolution in the region by 0.5 K. The relatively high spatial resolution of the data enabled a regionalization within the study area of 680 000 km2 into six sub‐regions based on principal component analysis of the monthly series. Long‐term temperature evolution proved to be highly similar across the region—thus making a mean series (averaged over all 97 single series) representative of the study area. Trend analysis (based on progressive forward and backward Mann–Kendall statistics and on progressive analysis of linear regression coefficients) was performed on seasonal and annual series. The results diverge from those of global datasets. This is mainly due to the extension of the 240‐year Alpine dataset by 100 years prior to the mid‐19th century, and also due to the advantages of a dense and homogenized regional dataset. The long‐term features include an initial decrease of the annual and seasonal series to a minimum followed by a positive trend until 1998. The minima are 1890 for the entire year and winter, 1840 for spring and 1920 for summer and autumn, respectively. The initial decreasing trend is more evident in spring and summer, less in autumn and smallest in winter. The mean annual temperature increase since 1890 in the Alps is 1.1 K, which is twice as much as the 0.55 K in the respective grid boxes of the most frequently used global dataset of the Climatic Research Unit (CRU), University of East Anglia. To enable an easier and more systematic handling of the dataset, these data have been interpolated to a 1°×1° longitude–latitude grid. The 105 low‐elevation and 16 high‐elevation grid point series are widely available without restrictions for scientific research and can be obtained from the authors. Copyright © 2001 Royal Meteorological Society