bluuebunny/crossref_metadata_2025 huggingface dataset:
Consequences of a novel caveolin‐3 mutation in a large German family
Authors: Fischer Dirk , Schroers Anja , Blümcke Ingmar , Urbach Horst , Zerres Klaus , Mortier Wilhelm , Vorgerd Matthias , Schröder Rolf
Published: January 2003

Mutations in the human caveolin‐3 gene (cav‐3) on chromosome 3p25 have been described in limb girdle muscular dystrophy, rippling muscle disease, hyperCKemia, and distal myopathy. Here, we describe the genetic, myopathological, and clinical findings in a large German family harboring a novel heterozygous mutation (GAC→GAA) in codon 27 of the cav‐3 gene. This missense mutation causes an amino acid change from asparagine to glutamate (Asp27Glu) in the N‐terminal region of the Cav‐3 protein, which leads to a drastic decrease of Cav‐3 protein expression in skeletal muscle tissue. In keeping with an autosomal dominant mode of inheritance, this novel cav‐3 mutation was found to cosegregate with neuromuscular involvement in the reported family. Ultrastructural analysis of Cav‐3–deficient muscle showed an abnormal folding of the plasma membrane as well as multiple vesicular structures in the subsarcolemmal region. Neurological examination of all nine subjects from three generations harboring the novel cav‐3 mutation showed clear evidence of rippling muscle disease. However, only two of these nine patients showed isolated signs of rippling muscle disease without muscle weakness or atrophy, whereas five had additional signs of a distal myopathy and two fulfilled the diagnostic criteria of a coexisting limb girdle muscular dystrophy. These findings indicate that mutations in the human cav‐3 gene can lead to different and overlapping clinical phenotypes even within the same family. Different clinical phenotypes in caveolinopathies may be attributed to so far unidentified modifying factors/genes in the individual genetic background of affected patients. Ann Neurol 2003
Blockers of sodium and calcium entry protect axons from nitric oxide‐mediated degeneration
Authors: Kapoor Raju , Davies Meirion , Blaker Paul A. , Hall Susan M. , Smith Kenneth J.
Published: January 2003

Axonal degeneration can be an important cause of permanent disability in neurological disorders in which inflammation is prominent, including multiple sclerosis and Guillain–Barré syndrome. The mechanisms responsible for the degeneration remain unclear, but it is likely that axons succumb to factors produced at the site of inflammation, such as nitric oxide (NO). We previously have shown that axons exposed to NO in vivo can undergo degeneration, especially if the axons are electrically active during NO exposure. The axons may degenerate because NO can inhibit mitochondrial respiration, leading to intraaxonal accumulation of Na+ and Ca2+ ions. Here, we show that axons can be protected from NO‐mediated damage using low concentrations of Na+ channel blockers, or an inhibitor of Na+/Ca2+ exchange. Our findings suggest a new strategy for axonal protection in an inflammatory environment, which may be effective in preventing the accumulation of permanent disability in patients with neuroinflammatory disorders. Ann Neurol 2003
The neurobiology, diagnosis, and treatment of narcolepsy
Authors: Scammell Thomas E.
Published: January 2003

Narcolepsy is a common cause of chronic sleepiness distinguished by intrusions into wakefulness of physiological aspects of rapid eye movement sleep such as cataplexy and hallucinations. Recent advances provide compelling evidence that narcolepsy may be a neurodegenerative or autoimmune disorder resulting in a loss of hypothalamic neurons containing the neuropeptide orexin (also known as hypocretin). Because orexin promotes wakefulness and inhibits rapid eye movement sleep, its absence may permit inappropriate transitions between wakefulness and sleep. These discoveries have considerably improved our understanding of the neurobiology of sleep and should foster the development of rational treatments for a variety of sleep disorders. Ann Neurol 2003
Axonal injury within language network in primary progressive aphasia
Authors: Catani Marco , Piccirilli Massimo , Cherubini Antonio , Tarducci Roberto , Sciarma Tiziana , Gobbi Gianni , Pelliccioli Gianpiero , Petrillo Stefania Maria , Senin Umberto , Mecocci Patrizia
Published: January 2003

Primary progressive aphasia (PPA) is characterized by an isolated progressive impairment of word use and comprehension reflecting the distribution of pathological processes within the left hemisphere. We used proton magnetic resonance spectroscopy (1H‐MRS) to study in vivo the integrity of axonal fibers connecting perisylvian language areas in 11 patients with PPA, 11 subjects with Alzheimer's disease, and 22 controls. Brain metabolites (N‐acetylaspartate, myoinositol, choline, creatine) were measured bilaterally within a volume of interest located in the central portion of the superior longitudinal fasciculus, a long associative bundle connecting Broca's area with Wernicke's area, and other language regions of the temporal lobe. In the PPA group, there was an asymmetrical N‐acetylaspartate to creatine ratio reduction compared with Alzheimer's disease and controls, with greater changes on the left side. The myoinositol to creatine ratio was increased in the PPA group bilaterally compared with controls. The choline to creatine ratio did not differ among the three groups. These results indicate an asymmetrical focal axonal injury within the language network in PPA. The marked difference in the distribution of N‐acetylaspartate to creatine between PPA and Alzheimer's disease suggests that proton magnetic resonance spectroscopy may help to differentiate between these two conditions. Ann Neurol 2003;53:000–000
Magnetic resonance imaging and neurosphere therapy of stroke in rat
Authors: Zhang Zheng Gang , Jiang Quan , Zhang Ruilan , Zhang Li , Wang Lei , Zhang Lijei , Arniego Polly , Ho Khang‐Loon , Chopp Michael
Published: January 2003

We intracisternally transplanted subventricular zone (SVZ) cells labeled by ferromagnetic particles into stroked rats. Migration of transplanted cells was non‐invasively tracked using magnetic resonance imaging (MRI). We found that transplanted cells selectively migrated towards the ischemic parenchyma at a mean speed of 65 ± 14.6 μm/hr in living rats. Migration of transplanted cells in the brain was also measured histopathologically. Rats transplanted with SVZ cells exhibited significant improvement of neurological function. Our data suggest that intracisternal transplantation of SVZ cells provides an avenue for cell therapy of stroke and that MRI can be used to track grafted cells in the brain. Ann Neurol 2003
Transient suppression of tinnitus by transcranial magnetic stimulation
Authors: Plewnia Christian , Bartels Mathias , Gerloff Christian
Published: January 2003

It has been proposed that tinnitus is associated with an irregular activation of the temporoparietal cortex. If this activity is a functionally relevant component of the tinnitus‐related network, a virtual temporary lesion of this area should result in transient reduction of tinnitus. To test this hypothesis, we applied 10Hz repetitive transcranial magnetic stimulation to eight scalp and four control positions in 14 patients with chronic tinnitus. Stimulation of left temporoparietal cortex significantly reduced tinnitus (Friedman analysis of variance, p < 0.05; compared with control), indicating that secondary auditory areas can be critical for tinnitus perception, perhaps as a consequence of maladaptive cortical reorganization. Ann Neurol 2003
Expression of cyclooxygenase‐2 in embryonic and fetal tissues during organogenesis and late pregnancy
Authors: Stanfield K.M. , Bell R.R. , Lisowski A.R. , English M.L. , Saldeen S.S. , Khan K. Nasir M.
Published: January 2003

BACKGROUND

Cyclooxygenase (COX) catalyzes the committed step in prostaglandin biosynthesis and exists as two related but unique isoforms, COX‐1 (constitutive) and COX‐2 (inducible). Prostaglandins (PGs) are known to have many important functions in reproduction, such as placentation and decidualization. Studies with the COX‐1 and COX‐2 knockout mice have demonstrated that COX‐2, but not COX‐1, is crucial for normal ovulation, implantation, and decidualization, suggesting that COX‐2–derived PGs are important during the initial stages of pregnancy. Although the COX‐2 knockout mice did not exhibit any abnormalities at birth, relatively little information exists with regard to the expression of COX‐2 in the fetus during development.

METHODS

In order to understand the role of COX‐2 throughout pregnancy, we characterized the cell type and the temporal expression of inducible COX‐2 throughout embryonic and fetal development in the rat (n = 22) by immunohistochemistry and in situ hybridization.

RESULTS

High levels of COX‐2 expression were seen in decidualized uterine tissue on gestation days 7–13 and then in the fetal membranes on gestation days 17–20. Cyclooxygenase‐2 expression was not detectable in any tissues from developing embryos during gestation days 7–13, but was observed in the fetal growth period (gestation days 15–20) in the skin, heart, cartilage, and the kidney.

CONCLUSIONS

No COX‐2 expression was seen in fetal tissues at days 7–13 of gestation, but was seen in various tissues at days 15–17 of gestation. These observations suggest that COX‐2 may be important in mid to late pregnancy through an effect on fetal organ growth, but not in the organogenetic phase of fetal development. Birth Defects Research (Part A) 67:54–58, 2003. © 2003 Wiley‐Liss, Inc.
Surface engineering of living myoblasts via selective periodate oxidation
Authors: De Bank P. A. , Kellam B. , Kendall D. A. , Shakesheff K. M.
Published: January 2003

Cell surface molecules are vital for normal cell activity. To study the functions of these molecules or manipulate cell behavior, the ability to decorate cell surfaces with bioactive molecules of our choosing is a potentially powerful technique. Here, we describe the molecular engineering of living L6 myoblast monolayers via selective periodate oxidation of sialic acid residues and the application of this surface modification in the artificial aggregation of cells. The aldehyde groups generated by this reaction were used to selectively ligate a model molecule, biotin hydrazide, to the cell surfaces. Flow cytometry analysis after staining with fluorescently conjugated avidin revealed a concentration‐dependent increase in fluorescence compared to untreated cells with a maximal shift of 345.1 ± 27.4‐fold and an EC50 of 17.4 ± 1.1 μM. This mild oxidation reaction did not affect cell number, viability, or morphology. We then compared this chemical technique with the metabolic incorporation of reactive cell surface ketone groups using N‐levulinoylmannosamine (ManLev). In this cell line, only a 22.3‐fold fluorescence shift was observed compared to untreated cells when myoblasts were incubated with a high concentration of ManLev for 48 hours. Periodate oxidation was then used to modify myoblast surfaces to induce cell aggregation. Crosslinking biotinylated myoblasts, which do not spontaneously aggregate in culture, with avidin resulted in the rapid formation of millimeter‐sized, multicellular structures. These data indicate that sodium periodate treatment is an effective, noncytotoxic method for the in vitro molecular engineering of living cell surfaces with the potential for cell biology and tissue engineering applications. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 800–808, 2003.
An EPR study of the secretion of G‐CSF heterologous protein from Pichia pastoris
Authors: Anžur Lasnik M. , Nemec M. , Romih R. , Schara M.
Published: January 2003

The biologically active protein known as human granulocyte colony stimulating factor (G‐CSF) can be efficiently secreted from the transformed GS115 Pichia pastoris (GS115/pPIC9/G‐CSF), which contains an α‐mating‐factor prepro signal sequence and an alcohol oxidase I promoter, both introduced using the pPIC9 plasmid. To study the P. pastoris G‐CSF protein‐secretion process, changes to the plasma membrane's lateral domain structure were monitored using electron paramagnetic resonance (EPR). The GS115 and its transformed analogue show that the plasma membrane can be described by fluid‐disordered and fluid‐ordered lateral domains. The relative proportion of these domains is a sensitive parameter that is able to describe the membrane's involvement in the protein‐excretion process. Here, P. pastoris GS115 served as a control for us to compare with the GS115/pPIC9/G‐CSF heterologous protein‐secreting cells. Electron paramagnetic resonance studies using the spin‐probe 5‐doxyl methyl ester of palmitic acid [MeFASL (10,3)] showed that during cultivation in a glycerol medium all types of cells had a relatively high proportion of cell‐membrane fluid‐disordered domains, while during the production phase the G‐CSF protein‐secreting cells showed a decrease in the proportion of fluid‐disordered domains. We ascribe this effect to the vesicular lipid exchange caused by the fusion of secretary vesicles with the plasma membrane during exocytosis. Using electron microscopy and immunocytochemistry intracellular vesicles containing the G‐CSF protein were detected. Our studies support the exocytotic mechanism of the heterologous protein secretion. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 768–774, 2003.
Quantitative comparison of transient growth of Saccharomyces cerevisiae, Saccharomyces kluyveri, and Kluyveromyces lactis
Authors: Herwig Christoph , von Stockar Urs
Published: January 2003

A multitude of metabolic regulations occur in yeast, particularly under dynamic process conditions, such as under sudden glucose excess. However, quantification of regulations and classification of yeast strains under these conditions have yet to be elucidated, which requires high‐frequency and consistent quantification of the metabolic response. The present study aimed at quantifying the dynamic regulation of the central metabolism of strains Saccharomyces cerevisiae, S. kluyveri, and Kluyveromyces lactis upon sudden glucose excess, accomplished by a shift‐up in dilution rate inside of the oxidative region using a small metabolic flux model. It was found that, under transient growth conditions, S. kluyveri behaved like K. lactis, while classification using steady‐state conditions would position S. kluyveri close to S. cerevisiae. For transient conditions and based on the observation whether excess glucose is initially used for catabolism (energy) or anabolism (carbon), we propose to classify strains into energy‐driven, such as S. cerevisiae, and carbon‐driven, such as S. kluyveri and K. lactis, strains. Furthermore, it was found that the delayed onset of fermentative catabolism in carbon‐driven strains is a consequence of low catabolic flux and the initial shunt of glucose in non‐nitrogen‐containing biomass constituents. The MFA model suggests that energy limitation forced the cell to ultimately increase catabolic flux, while the capacity of oxidative catabolism is not sufficient to process this flux oxidatively. The combination of transient experiments and its exploitation with reconciled intrinsic rates using a small metabolic model could corroborate earlier findings of metabolic regulations, such as tight glucose control in carbon‐driven strains and transient changes in biomass composition, as well as explore new regulations, such as assimilation of ethanol before glucose. The benefit from using small metabolic flux models is the richness of information and the enhanced insight into intrinsic metabolic pathways without a priori knowledge of adaptation kinetics. Used in an online context, this approach serves as an efficient tool for strain characterization and physiological studies. © 2003 Wiley Periodicals. Biotechnol Bioeng 81: 837–847, 2003.
Solid‐phase peptide synthesis by ion‐paired α‐chymotrypsin in nonaqueous media
Authors: Altreuter David H. , Dordick Jonathan S. , Clark Douglas S.
Published: January 2003

Solid‐phase synthesis of dipeptides in low‐water media was achieved using AOT ion‐paired α‐chymotrypsin solubilized in organic solvents. Multiple solvents and systematic variation of water activity, aw, were used to examine the rate of coupling between N‐α‐benzyloxycarbonyl‐L‐phenylalanine methyl ester (Z‐Phe‐OMe) and leucine as a function of the reaction medium for both solid‐phase and solution‐phase reactions. In solution, the observed maximum reaction rate in a given solvent generally correlated with measures of hydrophobicity such as the log of the 1‐octanol/water partitioning coefficient (log P) and the Hildebrand solubility parameter. The maximum rate for solution‐phase synthesis (13 mmol/h g‐enzyme) was obtained in a 90/10 (v/v) isooctane/tetrahydrofuran solvent mixture at an aw of 0.30. For the synthesis of dipeptides from solid‐phase leucine residues, the highest synthetic rates (0.14–1.3 mmol/h g‐enzyme) were confined to solvent environments that fell inside abruptly defined regions of solvent parameter space (e.g., log P > 2.3 and normalized electron acceptance index <0.13). The maximum rate for solid‐phase synthesis was obtained in a 90/10 (v/v) isooctane/tetrahydrofuran solvent mixture at an aw of 0.14. In 90/10 and 70/30 (v/v) isooctane/tetrahydrofuran environments with aw set to 0.14, seven different N‐protected dipeptides were synthesized on commercially available Tentagel support with yields of 74–98% in 24 h. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 809–817, 2003.
Self‐reflection and self‐practice in training of Cognitive Behaviour Therapy: an overview
Authors: Laireiter Anton‐Rupert , Willutzki Ulrike
Published: January 2003

The present paper deals with a developing training element in cognitive behaviour therapy (CBT), the self‐practice of therapeutic techniques and the self‐exploration of the person of the therapist. Initially, the current status of this training element in CBT is discussed, and a short note on its terminology is presented. Then an overview of the most important objectives of self‐practice and self‐reflection in CBT, and the concepts and methods of practice of this element is given. The concepts focus on two major aims, the self‐exploration of the person of the therapist within and beyond his or her therapeutic practice, and the self‐application of therapeutic techniques (= self‐practice). In the second part of the paper empirical studies related to the outcome of self‐reflection and self‐practice on the development of the therapist and her or his therapeutic practice are reviewed. The few empirical studies show that trainees experience in their own view substantial professional and personal gains from this training tool, with the professional impact being more important than the personal one. The most important outcomes evaluated by subjective data from the trainees are improvements in self‐insight and self‐awareness and a better understanding of the therapist's role and the therapeutic change process. Additionally, a better understanding of CBT methods and of general therapeutic skills, such as empathy and role‐taking‐competencies, are reported by the trainees. The paper comes to the conclusion that self‐exploration and self‐practice are important components of CBT training. Consequences and recommendations for its integration into training courses for CBT are discussed as well as the necessity for more research in this area. Copyright © 2002 John Wiley & Sons, Ltd.
Trainee clinical psychologists' adaptation and professional functioning: a longitudinal study
Authors: Kuyken Willem , Peters Emmanuelle , Power Michael J. , Lavender Tony
Published: January 2003

Clinical psychology training is a professionally formative period, which provides an opportunity to enable trainees to learn good self‐care skills and mature approaches to learning. If realized, this can support lifelong learning and high levels of professional functioning. This study sought to use transactional coping theory and experiential learning theory to improve our understanding of what factors predict changes in psychological adaptation and professional functioning over the course of clinical psychology training. A mixed prospective within‐persons and cross‐sectional between‐persons design was used. A sample of 183 trainee clinical psychologists (60.2% response rate) from 15 British clinical psychology training programmes participated at time one, 167 of whom participated at time 2 a year later (91.3% of the time 1 sample). They completed measures of appraisal, coping, social support and professional functioning at times 1 and 2. Path analyses suggested that trainees who appraised the demands of training as manageable, and reported greater access to appropriate support, engaged in less avoidance coping, reported fewer problems of psychological adaptation and were more likely to approach the task of learning and working appropriately and resiliently. Implications for clinical psychology training are suggested. Copyright © 2003 John Wiley & Sons, Ltd.
Identification of genes induced in regenerating Xenopus tadpole tails by using the differential display method
Authors: Ishino Tomoko , Shirai Mie , Kunieda Takekazu , Sekimizu Kazuhisa , Natori Shunji , Kubo Takeo
Published: January 2003

To identify candidate gene(s) involved in the tail regeneration of Xenopus laevis tadpoles, we used the differential display method to isolate four genes (clones 1, 2, 13a, and 13b) whose expression is induced in regenerating tadpole tails. Among them, clones 13a and 13b were found to encode the Xenopus homologues of the α1 chain of type XVIII collagen and neuronal pentraxin I, respectively. Expression of clone 2 and neuronal pentraxin I genes increased dramatically in the blastema 3 days after amputation, whereas that for the clone 1 and type XVIII collagen genes was induced gradually after amputation. In situ hybridization revealed that the neuronal pentraxin I gene is expressed specifically in the regenerating tail epidermis but not in the normal tail epidermis or the most distal margin of the tail blastema, suggesting that it has a tissue‐inductive role in tail regeneration. Expression of the four genes was induced in the limb and in the tail blastema, suggesting that they are involved in the regeneration of both organs. Finally, expression of clone 2 and neuronal pentraxin I genes was scarce during embryonic stages in comparison to the tail blastema, suggesting that their main functions are in organ regeneration. Our results demonstrate unique features of spatial and temporal gene expression patterns during Xenopus tadpole tail regeneration. Developmental Dynamics 226:317–325, 2003. © 2003 Wiley‐Liss, Inc.
Recruitment of postmitotic neurons into the regenerating spinal cord of urodeles
Authors: Zhang Fang , Ferretti Patrizia , Clarke Jonathan D.W.
Published: January 2003

By using fluorescent tracers, we have investigated the origin of the cells that form the regenerating spinal cord after tail amputation in urodele amphibians. We show that spinal cord cells immediately adjacent to the amputation plane die and are removed by phagocytic cells. Spinal cells just anterior to these dying cells are destined to make the majority of the regenerating cord. The largest contribution is likely to come from the radial ependymal cells, but we also demonstrate that postmitotic neurons in this location can translocate into the regenerating cord. These neurons integrate into the regenerate structure and survive for at least 4 weeks. We find no evidence that these translocated neurons dedifferentiate and divide during this regeneration process. We discuss the possibility that these neurons survive long term in the regenerate cord and become part of the functional neuronal circuitry. Developmental Dynamics 226:341–348, 2003. © 2003 Wiley‐Liss, Inc.
Bone formation during forelimb regeneration: A microtomography (microCT) analysis
Authors: Stock Stuart R. , Blackburn David , Gradassi Michael , Simon Hans‐Georg
Published: January 2003

In our study of bone regeneration in the forelimbs of mature newts (Notophthalmus viridescens), we used noninvasive X‐ray microtomography (microCT) to image regenerating limbs from 37 to 85 days and matching (contralateral) controls. We compared the patterns of regenerated and existing (nonregenerated) bone, investigating in particular the onset of mineralization of specific bones, the level of mineral present, and the lengths of the different bones. Overall, we find that the missing limb skeletal elements are restored in a proximal‐to‐distal direction, which reiterates the developmental patterning program. However, in contrast to this proximal–distal sequence, the portion of the humerus distal to the amputation site fails to ossify in synchrony with the regenerating radius and ulna. This finding suggests that the replacement of cartilage with mineralized bone close to the amputation site is delayed with respect to other regenerating skeletal elements. Developmental Dynamics 226:410–417, 2003. © 2003 Wiley‐Liss, Inc.
Mediolateral intercalation in planarians revealed by grafting experiments
Authors: Saito Yumi , Koinuma Satoshi , Watanabe Kenji , Agata Kiyokazu
Published: January 2003

We investigated how planarians organize their left–right axis by using ectopic grafting. Planarians have three body axes: anteroposterior (A‐P), dorsoventral (D‐V), and left–right (L‐R). When a small piece is implanted into an ectopic region along the A‐P and D‐V axes, intercalary structures are always formed to compensate for positional gaps. There are two hypotheses regarding L‐R axis formation in this organism: first, that the left and right sides of the animal may be recognized as different parts, and L‐R intercalation can induce midline structures (asymmetry hypothesis); second, that both sides may have symmetrical positional values, and mediolateral (M‐L) intercalation creates positional values along the L‐R axis (symmetry hypothesis). We performed ectopic grafting experiments in the head region of the planarian, Dugesia japonica, to examine these hypotheses. A left lateral fragment containing a left auricle was implanted into the medial region of the host. Ectopic structures were always formed only on the left side of the graft, where lateral tissues abutted onto the medial tissues. However, no morphologic change was induced on the right side of the graft, where left‐sided tissues faced onto right‐sided tissues. Molecular marker analyses indicated that ectopic structures formed on the left side of the graft were induced by M‐L intercalation, supporting the “symmetry hypothesis.” When the midline tissues were implanted into a lateral region, they induced a complete ectopic head, demonstrating that M‐L intercalation may be sufficient to establish the L‐R axis in planarians. Developmental Dynamics 226:334–340, 2003.© 2003 Wiley‐Liss, Inc.
Old questions, new tools, and some answers to the mystery of fin regeneration
Authors: Akimenko Marie‐Andrée , Marí‐Beffa Manuel , Becerra José , Géraudie Jacqueline
Published: January 2003

Pluridisciplinary approaches led to the notion that fin regeneration is an intricate phenomenon involving epithelial–mesenchymal and reciprocal exchanges throughout the process as well as interactions between ray and interray tissue. The establishment of a blastema after fin amputation is the first event leading to the reconstruction of the missing part of the fin. Here, we review our knowledge on the origin of the blastema, its formation and growth, and of the mechanisms that control differentiation and patterning of the regenerate. Our current understanding results from studies of fin regeneration performed in various teleost fish over the past century. We also report the recent breakthroughs that have been made in the past decade with the arrival of a new model, the zebrafish, Danio rerio, which now offers the possibility to combine cytologic, molecular, and genetic analyses and open new perspectives in this field. Developmental Dynamics 226:190–201, 2003. © 2003 Wiley‐Liss, Inc.
Intercalary regeneration in planarians
Authors: Agata K. , Tanaka T. , Kobayashi C. , Kato K. , Saitoh Y.
Published: January 2003

How can a planarian regenerate its entire body from a small portion of its body? Neoblasts, the totipotent stem cells of planarian, are assumed to be able to produce all missing cell types. However, we do not know how the cell fate of these cells is controlled during regeneration. Our recent studies with molecular markers suggest that intercalary regeneration is the fundamental principle in planarian regeneration. Here, we introduce the intercalation induced by ectopic grafting along the anteroposterior (A‐P), dorsoventral (D‐V), and left–right (L‐R) axes. Blastema formation is evoked by ectopic D‐V interactions after wound closure. Intercalation between the blastema and stump induces rearrangement of the positional identities along the A‐P axis. Consequently, totipotent stem cells change their differentiation patterns according to the newly rearranged positional identities along the A‐P, D‐V, and L‐R axes. According to the classic view, the blastema is regarded as the place where undifferentiated cells accumulate and regenerative events occur. Here, we propose a new interpretation, i.e., that the blastema may work as a signaling center inducing intercalary regeneration. Also, the roles of molecules and genes involved in intercalary regeneration are discussed. Developmental Dynamics 226:308–316, 2003. © 2003 Wiley‐Liss, Inc.
Rearrangements and increased expression of cyclin D1 (CCND1) in neuroblastoma
Authors: Molenaar Jan J. , van Sluis Peter , Boon Kathy , Versteeg Rogier , Caron Huib N.
Published: January 2003

Cyclin D1 regulates G1 cell cycle progression by controlling the phosphorylation of the retinoblastoma protein. This pathway is frequently deregulated in many malignancies. In neuroblastoma, however, no consistent G1 cell cycle checkpoint aberrations have been found. We examined the possible deregulation of cyclin D1 (CCND1) in this tumor. mRNA expression profiles of neuroblastoma generated by SAGE (Serial Analysis of Gene Expression) revealed a high expression of CCND1 in a subset of neuroblastoma cell lines and tumors. The CCND1 expression level can be 0.3% of the total cellular mRNA. Northern blot analysis of CCND1 expression showed a relative overexpression in 16 of 23 neuroblastoma cell lines and 10 of 15 tumor samples. In the majority of cases, the high CCND1 mRNA levels also led to high CCND1 protein levels. In the search for mechanisms causing this relative overexpression, we screened for amplifications and rearrangements of CCND1. Five amplifications were found in 202 neuroblastoma tumors and cell lines. Analysis of the 3′‐UTR of CCND1 showed a rearrangement in 1 of 96 tumors. These clonal aberrations of CCND1 together with the high expression suggest a role for deregulated CCND1 activity in neuroblastoma tumorigenesis. © 2003 Wiley‐Liss, Inc.
Cryptic chromosomal aberrations leading to an AML1/ETO rearrangement are frequently caused by small insertions
Authors: Gamerdinger Ulrike , Teigler‐Schlegel Andrea , Pils Sabine , Bruch Jochen , Viehmann Susanne , Keller Monika , Jauch Anna , Harbott Jochen
Published: January 2003

The translocation t(8;21)(q22;q22), which results in the fusion of the AML1 (RUNX1) and ETO (CBFA2T1) genes, is a recurrent aberration in acute myeloid leukemia (AML), preferentially correlated with FAB M2, and has the highest incidence in childhood AML. Because of the favorable prognosis, the evidence of the t(8;21) or the AML1/ETO fusion gene is mandatory in most of the therapy trials, allowing the stratification of the patients to the correct risk group in terms of treatment. Here we present six out of 59 children with AML who were positive for AML1/ETO by RT‐PCR, but showed no evidence of the classical t(8;21)(q22;q22) by conventional cytogenetics. Because of the discrepancies between molecular and cytogenetic analyses, these six patients were further investigated by fluorescence in situ hybridization analysis. Small hidden interstitial insertions resulting in an AML1/ETO rearrangement were detected in five (8.5%) of the 59 patients, whereas the sixth patient showed a cryptic three‐way translocation. The insertions could be characterized as ins(21;8) in three patients and ins(8;21) in the remaining two. Additionally, three of the patients showed secondary chromosome aberrations leading to a higher complexity of the karyotype. In conclusion, the combination of more than one standard technique in the analysis of AML1/ETO is useful to reveal the overall frequency of cryptic chromosome rearrangements and permits a better understanding of the mechanisms involved in the generation of this fusion gene. © 2003 Wiley‐Liss, Inc.
Lymphoma‐ and leukemia‐associated chromosomal translocations in healthy individuals
Authors: Janz Siegfried , Potter Michael , Rabkin Charles S.
Published: January 2003

Chromosomal translocations (CTs) are hallmark mutations of hematopoietic malignancy that result in the deregulated expression of oncogenes or the generation of novel fusion genes. The polymerase chain reaction (PCR) can be used to detect illegitimate recombinations of genomic DNA sequences as a more sensitive assay than cytogenetics for determining the presence of CTs. Both direct DNA‐PCR and reverse transcriptase‐PCR were used to examine healthy individuals for lymphoma‐ and leukemia‐associated CTs. Two oncogene‐activating CTs [t(14;18)(q32;q21) and t(8;14)(q24;q32)] and one fusion‐gene CT [t(2;5)(p23;q35)] from lymphomas and five fusion‐gene CTs from leukemia [t(9;22)(q34;q11), t(4;11)(q21;q23), t(15;17)(q22;q11), t(12;21)(p13;q22), t(8;21)(q22;q22)] were detected in such studies. The biological implication is that CTs associated with malignant tumors may also be found in cells that are not neoplastic. CTs are characteristic attributes of neoplastic clones but are by themselves insufficient to cause malignant transformation. A better understanding of the special biology of non‐neoplastic CT‐bearing cells will provide insight into their putative role as tumor precursors. Prospective epidemiological studies are needed to determine whether such cells in healthy individuals may, in some instances, become clonogenic founders of lymphoma or leukemia. © 2003 Wiley‐Liss, Inc.
Role of dimensionless numbers in wave analysis
Authors: Mishra Surendra Kumar , Singh Vijay P.
Published: January 2003

Dimensionless numbers, including the Reynolds number Re, the wave number $\hat{\sigma}$, the Froude number Fo, the quasi‐shallow wave number $\hat{\sigma}_{\rm v}$, the kinematic wave number K, Vedernikov number V, and others, are frequently encountered in analytical and numerical analyses of waves occurring in frictional and frictionless channels. Depending on their derivation, these numbers can be classified into physical, geometrical, derived, input‐based numbers, and their combinations. It can be shown that (a) the waves under friction are primarily governed by Re, pointing to friction laws and, consequently, the rating exponent; (b) the wave number characterizes the wave type for a given Re; (c) the numbers describing the reference state of flow do not have the efficacy of characterizing a wave type on its own, e.g. Fo that describes the significance of inertial forces in wave motion; (d) coupling of Fo with $\hat{\sigma}$ produces a more efficacious number K; (e) Vedernikov number V, a combination of the rating exponent or, in turn, Re and Fo, is more powerful than either Re or Fo; and (f) the numbers emanating from analytical solutions describe the wave types, whereas those emanating from numerical analyses usually prescribe the limits for stable solutions, such as Courant number Cr and cell Reynolds number D. The significance of these numbers is described using the case of evolution and propagation of a dam‐break flood wave. Furthermore, it is shown that as $\hat{\sigma}$ forms the basic unit of shallow wave description, then quasi‐shallow waves are described by the quasi‐shallow wave number $\hat{\sigma}_{\rm v}$, the wave length parameter normalized by the normal depth of flow. Copyright © 2002 John Wiley & Sons, Ltd.
Delineating protein–protein interactions via biomolecular interaction analysis–mass spectrometry
Authors: Nedelkov Dobrin , Nelson Randall W.
Published: January 2003

The utility of biomolecular interaction analysis–mass spectrometry (BIA/MS) in screening for protein–protein interactions was explored in this work. Experiments were performed in which proteins served as ligands for screening of possible interactions with other proteins from human plasma and urine. The proteins utilized were beta‐2‐microglobulin, cystatin C (cysC), retinol binding protein (RBP), transthyretin (TTR), alpha‐1‐microglobulin, C‐reactive protein, transferrin and papain. The immobilization of functionally active proteins was confirmed via interactions with antibodies to the corresponding proteins. Various dilutions of human urine and plasma were injected over the protein‐derivatized surfaces. It was observed that the urine injections generally yielded smaller SPR responses than those observed after the plasma injections. The BIA/MS experiments did not reveal novel protein–protein interactions, although several established interactions (such as those between RBP and TTR, and cysC and papain) were validated. Few protein ligand deficiencies (such as truncations) leading to false negative and false positive BIA/MS results were also discovered. Copyright © 2003 John Wiley & Sons, Ltd.
BEPITOPE: predicting the location of continuous epitopes and patterns in proteins
Authors: Odorico Michael , Pellequer Jean‐Luc
Published: January 2003

In growing need of obtaining highly specific monoclonal antibodies against novel proteins, we developed new functions implemented in the program BEPITOPE to predict continuous protein epitopes. This program not only can compute, combine, display and print prediction profiles, but also provides a list of suggested linear peptides to be synthesized. Novel facilities incorporated in BEPITOPE include the treatment of a whole genome, the search for a user‐defined pattern, and the combination of prediction to pattern profiles. This latter approach is useful to remove unwanted predictions such as those including glycosylation sites. Copyright © 2003 John Wiley & Sons, Ltd.
Interaction of epitope‐related and ‐unrelated peptides with anti‐p24 (HIV‐1) monoclonal antibody CB4‐1 and its Fab fragment
Authors: Welfle Karin , Misselwitz Rolf , Höhne Wolfgang , Welfle Heinz
Published: January 2003

The binding of four epitope‐related peptides and three library‐derived, epitope‐unrelated peptides of different lengths (10–14 amino acids) and sequence by anti‐p24 (HIV‐1) monoclonal antibody CB4‐1 and its Fab fragment was studied by isothermal titration calorimetry. The binding constants KA at 25 °C vary between 5.1 × 107 M
−1 for the strongest and 1.4 × 105 M
−1 for the weakest binder. For each of the peptides complex formation is enthalpically driven and connected with unfavorable entropic contributions; however, the ratio of enthalpy and entropy contributions to ΔG0 differs markedly for the individual peptides. A plot of −ΔH0 vs −TΔS0 shows a linear correlation of the data for a wide variety of experimental conditions as expected for a process with ΔCp much larger than ΔS0. The dissimilarity of ΔCp and ΔS0 also explains why ΔH0 and TΔS0 show similar temperature dependences resulting in relatively small changes of ΔG0 with temperature. The heat capacity changes ΔCp upon antibody–peptide complex formation determined for three selected peptides vary only in a small range, indicating basic thermodynamic similarity despite different key residues interacting in the complexes. Furthermore, the comparison of van't Hoff and calorimetric enthalpies point to a non‐two‐state binding mechanism. Protonation effects were excluded by measurements in buffers of different ionization enthalpies. Differences in the solution conformation of the peptides as demonstrated by circular dichroic measurements do not explain different binding affinities of the peptides; specifically a high helix content in solution is not essential for high binding affinity despite the helical epitope conformation in the crystal structure of p24. Copyright © 2003 John Wiley & Sons, Ltd.
Seasonal modulation of the El Niño–southern oscillation relationship with sea level pressure anomalies over the North Atlantic in October–March 1873–1996
Authors: Moron V. , Gouirand I.
Published: January 2003

The seasonal modulation of the relationship between the sea‐level pressure anomalies (SLPAs) over the North Atlantic region (100°W–50°E; 20–70°N) and the sea surface temperature anomalies (SSTAs) in the tropical Pacific (120–290°E; 20°N–20°S) is investigated in the northern winter (October to March) from 1873 to 1996, using singular‐value decomposition and composite analyses. Both methods show that the pattern of the North Atlantic SLPA associated with the tropical Pacific SSTA in November–December is different from that found in January–March. The surface covered by a significant SLPA is larger in November–December and February–March than in January. In November–December, the warm El Niño–southern oscillation (ENSO) events are associated with negative SLPAs extending from the Hudson Bay to Scandinavia and positive SLPAs over the Azores high. The cold ENSO events are associated with a positive SLPA between Greenland and western Europe. In January, and mainly in February–March, the warm ENSO events are associated with a positive SLPA north of 50°N and a negative SLPA extending from the southeastern USA toward western and central Europe. The cold ENSO events exhibit almost reversed SLPA patterns. The change between November–December and January–March is also observed at the hemispheric scale. In November–December, the SLPAs associated with the warm minus cold ENSO composite form a hemispheric north–south dipole pattern with positive (negative) anomalies south (north) of 40–45°N. In January–March, the SLPA pattern associated with the warm minus cold ENSO composite is close to the tropical Northern‐Hemisphere pattern. Copyright © 2003 Royal Meteorological Society.
Spatial and temporal comparisons of droughts over Korea with East Asia
Authors: Min Seung‐Ki , Kwon Won‐Tae , Park E‐Hyung , Choi Youngeun
Published: January 2003

To investigate the spatial and temporal relationships of drought occurrence and intensity between Korea and East Asia, the Standardized Precipitation Index (SPI), calculated from Climatic Research Unit (CRU) monthly precipitation data, was used from 1951 to 1996. It is found that the frequency of occurrence of droughts in Korea has significant time intervals of 2–3 and 5–8 years and has been increasing since the 1980s. Correlation and composite analyses showed that the occurrence of droughts over central eastern China, Manchuria, and the north coast of Japan was highly correlated with those in Korea. However, the time scales of occurrence of droughts over the three regions were different. Droughts in eastern China represented in‐phase variations with those in Korea with a time interval of 5–8 years, whereas those in Manchuria occurred with a time interval of 15 years, and those in Japan had no coincident variations.

To assess the feasibility of usage of proxy climate data in eastern China for the research of droughts in Korea during the pre‐instrumental period, dry–wet indices in six regions of eastern China were correlated to the SPI values in Korea for the period of 1951–92 considering several time scales. Dry–wet indices of region V showed a larger correlation and the most similar trend to the droughts in Korea with a time scale of 5–11 years. This suggested an effective utilization of historical records and other proxy data in eastern China to understand extreme climatic events in Korea for the past 500 years. Copyright © 2003 Royal Meteorological Society.
An investigation of the physical and socioeconomic determinants of soil erosion in the Hararghe Highlands, eastern Ethiopia
Authors: Daba S.
Published: January 2003

The vicious poverty–environmental spiral commonly exists due to the interconnectedness of the socioeconomic aspects of farmers and land degradation. The socioeconomic situation of farmers affects their capabilities to implement environmentally viable soil and water conservation measures. These situations include farm practices and attitudes toward rational use of resources. An observational study was conducted to have an insight of the perception of farmers about the danger of gully erosion and their willingness to adopt new improved soil and water conservation measures. The variables studied included literacy, location of farm within the landscape, major land use, the severity of water scarcity, means of traction, perception, type of soil and water conservation measure in use, willingness to adopt new soil and water conservation measures and type of incentives needed for adopting new soil and water conservation measures. The variables studied were coded using dummy variable coding techniques and were subjected to rigorous statistical analysis using canonical correlation, logistic regression and discriminant analysis. Results suggest that perception about the danger of gully erosion is significantly correlated to severity of water scarcity, location of farm within the landscape and literacy of farmers. Willingness to adopt a new or improved soil and water conservation measure is strongly related to fertilizer availability as an incentive and to the literacy of the farmers. It is concluded that under Hararghe Highland conditions, design and implementation of soil and water conservation measures should take into account farmers' priorities such as alleviation of the water scarcity problem. Moreover, successful implementation of an improved or new measure of soil and water conservation measure is contingent upon the availability of incentives, primarily fertilizers. Copyright © 2002 John Wiley & Sons, Ltd.
Rural household responses to fuelwood scarcity in Nyando District, Kenya
Authors: Mahiri I. O.
Published: January 2003

The fuelwood debate over supply–demand balances is well rehearsed; and the use of simplistic linear models to explain supply–demand differentials hardly captures the complex patterns of use and dynamics of fuelwood in the rural household. This paper shows that households in Nyando District have evolved sophisticated local response mechanisms and strategies in coping with the fuelwood scarcity. Data collected through a multi‐method approach revealed, among others, a higher percentage of planted trees in Kochogo than in Awasi study sites. Despite this high proliferation of trees, rural households in Kochogo still identified fuelwood scarcity as a growing problem. The seeming ‘abundance’ of trees is not synonymous with the supply of fuelwood, or the alleviation of fuelwood scarcity. Rural households in Kochogo therefore resort to the market to purchase fuelwood, as well as adapt various fuel‐saving strategies and mechanisms to cope with the apparent scarcity. The purchase of crop residues from the market is a strong indicator of this scarcity, being simply lack of access to or entitlement to trees. The situation is different in Awasi, where there is a higher percentage of natural trees and clump bushes, which provide a relative abundance of fuelwood. This condition has encouraged local households not to keep a stock of fuelwood, but simply to collect from nearby bushes when required. Copyright © 2002 John Wiley & Sons, Ltd.