bluuebunny/crossref_metadata_2025 huggingface dataset:
Three new allelic mouse mutations that cause skeletal overgrowth involve the natriuretic peptide receptor C gene (
Npr3
)
Authors: Jaubert Jean , Jaubert Francis , Martin Natalia , Washburn Linda L. , Lee Barbara K. , Eicher Eva M. , Guénet Jean-Louis
Published: July 2002

In 1979, a BALB/cJ mouse was identified with an exceptionally long body. This phenotype was found to be caused by a recessive mutation, designated longjohn (
lgj
), that mapped to the proximal region of chromosome 15. Several years later, a mouse with a similarly elongated body was identified in an outbred stock after chemical mutagenesis with ethylnitrosourea. This phenotype also was caused by a recessive mutation, designated strigosus (
stri
). The two mutations were found to be allelic. A third allele was identified in a DBA/2J mouse and was designated longjohn-2J (
lgj
2J
). Analysis of skeletal preparations of
stri/stri
mice indicated that the endochondral ossification process was slightly delayed, resulting in an extended proliferation zone. A recent study reported that mice overexpressing brain natriuretic peptide, one of the members of the natriuretic peptide family, exhibit a skeletal-overgrowth syndrome with endochondral ossification defects. The
Npr3
gene coding for type C receptor for natriuretic peptides (NPR-C), which is mainly involved in the clearance of the natriuretic peptides, mapped in the vicinity of our mouse mutations and thus was a candidate gene. The present study reports that all three mutations involve the
Npr3
gene and provides evidence
in vivo
that there is a natriuretic-related bone pathway, underscoring the importance of natriuretic peptide clearance by natriuretic peptide type C receptor.
Acylation stabilizes a protease-resistant conformation of protoporphyrinogen oxidase, the molecular target of diphenyl ether-type herbicides
Authors: Arnould Sylvain , Takahashi Masayuki , Camadro Jean-Michel
Published: July 2002

Protein acylation is an important way in which a number of proteins with a variety of functions are modified. The physiological role of the acylation of cellular proteins is still poorly understood. Covalent binding of fatty acids to nonintegral membrane proteins is thought to produce transient or permanent enhancement of the association of the polypeptide chains with biological membranes. In this paper, we investigate the functional role for the palmitoylation of an atypical membrane-bound protein, yeast protoporphyrinogen oxidase, which is the molecular target of diphenyl ether-type herbicides. Palmitoylation stabilizes an active heat- and protease-resistant conformation of the protein. Palmitoylation of protoporphyrinogen oxidase has been demonstrated to occur
in vivo
both in yeast cells and in a heterologous bacterial expression system, where it may be inhibited by cerulenin leading to the accumulation of degradation products of the protein. The thiol ester linking palmitoleic acid to the polypeptide chain was shown to be sensitive to hydrolysis by hydroxylamine and also by the widely used serine-protease inhibitor phenylmethylsulfonyl fluoride.
Cloning, expression, and genetic mapping of Sema W, a member of the semaphorin family
Authors: Encinas Jeffrey A. , Kikuchi Kaoru , Chedotal Alain , de Castro Fernando , Goodman Corey S. , Kimura Toru
Published: July 2002

The semaphorins comprise a large family of membrane-bound and secreted proteins, some of which have been shown to function in axon guidance. We have cloned a transmembrane semaphorin, Sema W, that belongs to the class IV subgroup of the semaphorin family. The mouse and rat forms of Sema W show 97% amino acid sequence identity with each other, and each shows about 91% identity with the human form. The gene for Sema W is divided into 15 exons, up to 4 of which are absent in the human cDNAs that we sequenced. Unlike many other semaphorins, Sema W is expressed at low levels in the developing embryo but was found to be expressed at high levels in the adult central nervous system and lung. Functional studies with purified membrane fractions from COS7 cells transfected with a Sema W expression plasmid showed that Sema W has growth-cone collapse activity against retinal ganglion-cell axons, indicating that vertebrate transmembrane semaphorins, like secreted semaphorins, can collapse growth cones. Genetic mapping of human
SEMAW
with human/hamster radiation hybrids localized the gene to chromosome 2p13. Genetic mapping of mouse
Semaw
with mouse/hamster radiation hybrids localized the gene to chromosome 6, and physical mapping placed the gene on bacteria artificial chromosomes carrying microsatellite markers
D6Mit70
and
D6Mit189
. This localization places
Semaw
within the locus for motor neuron degeneration 2, making it an attractive candidate gene for this disease.
Channel-mediated high-affinity K
+
uptake into guard cells from
Arabidopsis
Authors: Brüggemann Lioubov , Dietrich Petra , Becker Dirk , Dreyer Ingo , Palme Klaus , Hedrich Rainer
Published: July 2002

Potassium uptake by higher plants is the result of high- or low-affinity transport accomplished by different sets of transporters. Although K
+
channels were thought to mediate low-affinity uptake only, the molecular mechanism of the high-affinity, proton-dependent K
+
uptake system is still scant. Taking advantage of the high-current resolution of the patch-clamp technique when applied to the small
Arabidopsis thaliana
guard cells densely packed with voltage-dependent K
+
channels, we could directly record channels working in the concentration range of high-affinity K
+
uptake systems. Here we show that the K
+
channel KAT1 expressed in
Arabidopsis
guard cells and yeast is capable of mediating potassium uptake from media containing as little as 10 μM of external K
+
. Upon reduction of the external K
+
content to the micromolar level the voltage dependence of the channel remained unaffected, indicating that this channel type represents a voltage sensor rather than a K
+
-sensing valve. This behavior results in K
+
release through K
+
uptake channels whenever the Nernst potential is negative to the activation threshold of the channel. In contrast to the H
+
-coupled K
+
symport shown to account for high-affinity K
+
uptake in roots, pH-dependent K
+
uptake into guard cells is a result of a shift in the voltage dependence of the K
+
channel. We conclude that plant K
+
channels activated by acid pH may play an essential role in K
+
uptake even from dilute solutions.
A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage
Authors: Brown Alexandra L. , Lee Chang-Hun , Schwarz Julie K. , Mitiku Nesanet , Piwnica-Worms Helen , Chung Jay H.
Published: July 2002

Checkpoints maintain the order and fidelity of the eukaryotic cell cycle, and defects in checkpoints contribute to genetic instability and cancer. Much of our current understanding of checkpoints comes from genetic studies conducted in yeast. In the fission yeast
Schizosaccharomyces pombe
(Sp), SpRad3 is an essential component of both the DNA damage and DNA replication checkpoints. The SpChk1 and SpCds1 protein kinases function downstream of SpRad3. SpChk1 is an effector of the DNA damage checkpoint and, in the absence of SpCds1, serves an essential function in the DNA replication checkpoint. SpCds1 functions in the DNA replication checkpoint and in the S phase DNA damage checkpoint. Human homologs of both SpRad3 and SpChk1 but not SpCds1 have been identified. Here we report the identification of a human cDNA encoding a protein (designated HuCds1) that shares sequence, structural, and functional similarity to SpCds1. HuCds1 was modified by phosphorylation and activated in response to ionizing radiation. It was also modified in response to hydroxyurea treatment. Functional ATM protein was required for HuCds1 modification after ionizing radiation but not after hydroxyurea treatment. Like its fission yeast counterpart, human Cds1 phosphorylated Cdc25C to promote the binding of 14-3-3 proteins. These findings suggest that the checkpoint function of HuCds1 is conserved in yeast and mammals.
Herpes simplex virus vector-mediated expression of Bcl-2 prevents 6-hydroxydopamine-induced degeneration of neurons in the substantia nigra
in vivo
Authors: Yamada Masanobu , Oligino Thomas , Mata Marina , Goss James R. , Glorioso Joseph C. , Fink David J.
Published: July 2002

6-Hydroxydopamine (6-OHDA) is widely used to selectively lesion dopaminergic neurons of the substantia nigra (SN) in the creation of animal models of Parkinson’s disease.
In vitro
, the death of PC-12 cells caused by exposure to 6-OHDA occurs with characteristics consistent with an apoptotic mechanism of cell death. To test the hypothesis that apoptotic pathways are involved in the death of dopaminergic neurons of the SN caused by 6-OHDA, we created a replication-defective genomic herpes simplex virus-based vector containing the coding sequence for the antiapoptotic peptide Bcl-2 under the transcriptional control of the simian cytomegalovirus immediate early promoter. Transfection of primary cortical neurons in culture with the Bcl-2-producing vector protected those cells from naturally occurring cell death over 3 weeks. Injection of the Bcl-2-expressing vector into SN of rats 1 week before injection of 6-OHDA into the ipsilateral striatum increased the survival of neurons in the SN, detected either by retrograde labeling of those cells with fluorogold or by tyrosine hydroxylase immunocytochemistry, by 50%. These results, demonstrating that death of nigral neurons induced by 6-OHDA lesioning may be blocked by the expression of Bcl-2, are consistent with the notion that cell death in this model system is at least in part apoptotic in nature and suggest that a Bcl-2-expressing vector may have therapeutic potential in the treatment of Parkinson’s disease.
Estrogen-induced activation of mitogen-activated protein kinase requires mobilization of intracellular calcium
Authors: Improta-Brears Teresa , Whorton A. Richard , Codazzi Franca , York John D. , Meyer Tobias , McDonnell Donald P.
Published: July 2002

Estrogens and growth factors such as epidermal growth factor (EGF) act as mitogens promoting cellular proliferation in the breast and in the reproductive tract. Although it was considered originally that these agents manifested their mitogenic actions through separate pathways, there is a growing body of evidence suggesting that the EGF and estrogen-mediated signaling pathways are intertwined. Indeed, it has been demonstrated recently that 17β-estradiol (E2) can induce a rapid activation of mitogen-activated protein kinase (MAPK) in mammalian cells, an event that is independent of both transcription and protein synthesis. In this study, we have used a pharmacological approach to dissect this novel pathway in MCF-7 breast cancer cells and have determined that in the presence of endogenous estrogen receptor, activation of MAPK by E2 is preceded by a rapid increase in cytosolic calcium. The involvement of intracellular calcium in this process was supported by the finding that the presence of EGTA and Ca
2+
-free medium did not affect the activation of MAPK by E2 and, additionally, that this response was blocked by the addition of the intracellular calcium chelator 1,2-bis(2-aminophenoxy)ethane-
N,N,N′,N′
-tetraacetate. Cumulatively, these data indicate that the estrogen receptor, in addition to functioning as a transcription factor, is also involved, through a nongenomic mechanism, in the regulation of both intracellular calcium homeostasis and MAPK-signaling pathways. Although nongenomic actions of estrogens have been suggested by numerous studies in the past, the ability to link estradiol and the estrogen receptor to a well defined signaling pathway strongly supports a physiological role for this activity.
1,2,3 = 2,3,4: A solution to the problem of the homology of the digits in the avian hand
Authors: Wagner Günter P. , Gauthier Jacques A.
Published: July 2002

Persistent contradictions in well supported empirical findings usually point to important scientific problems and may even lead to exciting new insights. One of the most enduring problems in evolutionary biology is the apparent conflict between paleontological and embryological evidence regarding the homology of the digits in the avian hand (1, 2). We propose that this problem highlights an important feature of morphological change: namely, the possible dissociation between the developmental origin of a particular repeated element and its subsequent individualization into a fully functional character. We argue that, although comparative embryological evidence correctly identifies the homology of the primordial condensations in avians as CII, CIII, and CIV, subsequent anatomical differentiation reflects a frame shift in the developmental identities of the avian digit anlagen in later ontogeny such that CII becomes DI, CIII becomes DII, and CIV becomes DIII.
Neonatal lead exposure impairs development of rodent barrel field cortex
Authors: Wilson Mary Ann , Johnston Michael V. , Goldstein Gary W. , Blue Mary E.
Published: July 2002

Childhood exposure to low-level lead can permanently reduce intelligence, but the neurobiologic mechanism for this effect is unknown. We examined the impact of lead exposure on the development of cortical columns, using the rodent barrel field as a model. In all areas of mammalian neocortex, cortical columns constitute a fundamental structural unit subserving information processing. Barrel field cortex contains columnar processing units with distinct clusters of layer IV neurons that receive sensory input from individual whiskers. In this study, rat pups were exposed to 0, 0.2, 1, 1.5, or 2 g/liter lead acetate in their dam's drinking water from birth through postnatal day 10. This treatment, which coincides with the development of segregated columns in the barrel field, produced blood lead concentrations from 1 to 31 μg/dl. On postnatal day 10, the area of the barrel field and of individual barrels was measured. A dose-related reduction in barrel field area was observed (Pearson correlation = −0.740;
P
< 0.001); mean barrel field area in the highest exposure group was decreased 12% versus controls. Individual barrels in the physiologically more active caudoventral group were affected preferentially. Total cortical area measured in the same sections was not altered significantly by lead exposure. These data support the hypothesis that lead exposure may impair the development of columnar processing units in immature neocortex. We demonstrate that low levels of blood lead, in the range seen in many impoverished inner-city children, cause structural alterations in a neocortical somatosensory map.
Trans-splicing vectors expand the utility of adeno-associated virus for gene therapy
Authors: Yan Ziying , Zhang Yulong , Duan Dongsheng , Engelhardt John F.
Published: July 2002

Adeno-associated viral (AAV) vectors have demonstrated considerable promise for gene therapy of inherited diseases. However, with a packaging size of <5 kb, applications have been limited to relatively small disease genes. Based on the finding that AAV genomes undergo intermolecular circular concatamerization after transduction in muscle, we have developed a paradigm to increase the size of delivered transgenes with this vector through trans-splicing between two independent vectors coadministered to the same tissue. When two vectors encoding either the 5′ or 3′ portions of the erythropoietin genomic locus were used, functional erythropoietin protein was expressed in muscle subsequent to the formation of intermolecular circular concatamers in a head-to-tail orientation through trans-splicing between these two independent vector genomes. These findings will allow for the application of AAV technologies to a wider variety of diseases for which therapeutic transgenes exceed the packaging limitation of present AAV vectors.
A cGMP-signaling pathway in a subset of olfactory sensory neurons
Authors: Meyer Mike R. , Angele Albert , Kremmer Elisabeth , Kaupp U. Benjamin , Müller Frank
Published: July 2002

It is well established that signal transduction in sensory neurons of the rat olfactory epithelium involves a cAMP-signaling pathway. However, a small number of olfactory neurons specifically express cGMP-signaling components, namely a guanylyl cyclase (GC-D) and a cGMP-stimulated phosphodiesterase (PDE2). Here, we show that this subset of olfactory neurons expressing GC-D and PDE2 does also express the subunit of a cGMP-selective cyclic nucleotide-gated (CNG) channel that has been previously identified in cone photoreceptors. Further, components of the prototypical cAMP-signaling pathway could not be detected in this subpopulation of cells. These results imply that these neurons use an alternative signaling pathway, with cGMP as the intracellular messenger, and that, in these cells, the receptor current is initiated by the opening of cGMP-gated channels.
An alternative interpretation of nanobacteria-induced biomineralization
Authors: Cisar John O. , Xu De-Qi , Thompson John , Swaim William , Hu Lan , Kopecko Dennis J.
Published: July 2002

The reported isolation of nanobacteria from human kidney stones
raises the intriguing possibility that these microorganisms are
etiological agents of pathological extraskeletal calcification
[Kajander, E. O. & Çiftçioglu, N. (1998)
Proc.
Natl. Acad. Sci. USA
95, 8274–8279]. Nanobacteria were
previously isolated from FBS after prolonged incubation in DMEM. These
bacteria initiated biomineralization of the culture medium and were
identified in calcified particles and biofilms by nucleic acid stains,
16S rDNA sequencing, electron microscopy, and the demonstration of a
transferable biomineralization activity. We have now identified
putative nanobacteria, not only from FBS, but also from human saliva
and dental plaque after the incubation of 0.45-μm membrane-filtered
samples in DMEM. Although biomineralization in our “cultures” was
transferable to fresh DMEM, molecular examination of decalcified
biofilms failed to detect nucleic acid or protein that would be
expected from growth of a living entity. In addition, biomineralization
was not inhibited by sodium azide. Furthermore, the 16S rDNA sequences
previously ascribed to
Nanobacterium sanguineum
and
Nanobacterium
sp. were found to be indistinguishable
from those of an environmental microorganism,
Phyllobacterium
mysinacearum
, that has been previously detected as a
contaminant in PCR. Thus, these data do not provide plausible support
for the existence of a previously undiscovered bacterial genus.
Instead, we provide evidence that biomineralization previously
attributed to nanobacteria may be initiated by nonliving macromolecules
and transferred on “subculture” by self-propagating
microcrystalline apatite.
Identification des facteurs liés à la consommation de cigarettes, d'alcool et de drogues à l'adolescence
Authors: Gosselin Catherine , Larocque Denis , Vitaro Frank , Gagnon Claude
Published: July 2002

Cette étude vise à identifier les facteurs d'ordre personnel, familial, social et scolaire liés à la consommation de substances psychotropes à l'adolescence. Environ 1000 adolescents ont participé à quatre collectes de données échellonnées sur une période de 44 mois. Les résultats montrent un lien entre la consommation précoce de substances psychotropes et la persistance de cette habitude. D'autres facteurs tels les compétences à entrer en interaction avec les pairs, l'inadaptation scolaire et le manque de supervision parentale sont également des prédicteurs d'une telle habitude. En retour, l'importance accordée aux qualités morales et les réactions négatives vis‐à‐vis les habitudes de consommation de la mère sont associées à un fléchissement de cette habitude. La discussion porte sur l'importance d'identifier les trajectoires développementales menant à la dépendance aux substances psychotropes.
L'accession à l'opérativité formelle chez les enfants intellectuellement précoces
Authors: Planche Pascale , Gicquel Marie‐Cécile
Published: July 2002

Cette recherche tente d'apporter des éléments d'information sur le fonctionnement et le développement cognitifs de l'enfant intellectuellement précoce, au seuil de l'opérativité formelle. Elle s'inscrit dans le cadre théorique du développement de l'intelligence selon Piaget. Des enfants “précoces” et des enfants “moyens”, d'âge mental équivalent (12 ans et 6 mois) ont été comparés. Un second groupe de précoces de même âge chronologique que les moyens, complète cette investigation. L'Échelle de Développement de la Pensée Logique de Longeot a été présentée à tous les sujets. Les résultats révèlent des aspects dyssynchroniques au sein de la genèse des enfants précoces. Ils sont discutés en prenant en considération les caractéristiques des tâches proposées (le type de raisonnement qu'elles imposent) et les spécificités fonctionnelles des sujets comparés.
Meeting New Challenges in Educational Research Training: The Signposts for Educational Research CD‐ROM
Authors: Barrett Elizabeth , Lally Vic
Published: July 2002

This article suggests that the professional educational research community is facing new challenges in the development of educational research training. In particular it argues that as the learning needs of the student population become more differentiated, there is a strong pressure to develop and evaluate the use of information and communications technologies (ICTs) to support research training in education. The article describes one response to this pressure: the development and evaluation of an interactive multimedia (IMM) resource, the Signposts for Educational Research CD‐ROM. The CD‐ROM is designed to be used by postgraduate students on a wide range of courses but is particularly aimed at those enrolled on part‐time and distance education programmes. Typically, part‐time students studying at a distance from their providing institution use print‐based materials in combination with occasional attendance at residential study schools; such residential schools frequently represent the only occasions on which tutors and students come together. Many distance learners therefore have limited contact with their tutors and peers, and only limited access to site‐based resources. At the same time, because of their different prior experiences of formal academic study, there is considerable diversity in the individual needs of students registered on part‐time and distance education programmes. This distinctive student profile indicates highly differentiated training requirements and thus the need for personal and customised support for research training. One of the reasons for producing the Signposts CD‐ROM was the perceived need to offer such students alternative modes of learning support. It is argued that the flexible structure of IMM materials may be effective in meeting the individual needs of students while the use of affective material (in the form of a ‘peer voice’) may help to build the confidence, and lessen the isolation, of the distance learner. The article concludes with a discussion of the impact on learning of IMM resources. In particular, it asks whether research students can ‘become critical’ in a multimedia environment, and attempts to assess some of the methodological difficulties involved in evaluating the use of IMM to support learning in this context.
Induction of the pro‐myelocytic leukaemia gene by type I and type II interferons
Authors: Heuser M. , van der Kuip H. , Falini B. , Peschel C. , Huber C. , Fischer T.
Published: July 2002

The physiological role of the pro‐myelocytic leukaemia (PML) gene product is poorly defined. Among other functions, PML is involved in haem atopoietic differentiation and in control of cell growth and tumorigenesis. We investigated the regulation of human PML expression by interferons (IFNs) and IL‐1 in various human haematopoietic lines (U937, THP1, HL60, NB4), in human diploid fibroblasts and in human peripheral blood leukocytes. Cytokineinduced modulation of PML expression was assessed by Northern blot analyses, flow cytometry studies and in situ immunolabelling. Our data show that IFNs and IL‐1 upregulate PML transcript and protein expression in a time and dose‐dependent manner. In situ immunolabelling revealed that upregulation of protein expression by IFN‐α is a consequence of a marked increase in both the number and the intensity of the staining of so‐called PML nuclear bodies. Our data suggest that stimulation of PML expression by interferons and IL‐1 may account for upregulation of PMLproteins observed in inflammatory tissues and in proliferative states.
Gender and Psychiatric Comorbidity: Impact on Clinical Presentation of Alcohol Dependence
Authors: Pettinati Helen M. , Rukstalis Margaret R. , Luck Gary J. , Volpicelli Joseph R. , O'Brien Charles P.
Published: July 2002

We examined differences in clinical presentation for outpatient alcohol treatment in: 1) males and females, considering comorbidity; and 2) three comorbid groups, consideringgender. Drinkingindices and emotional, physical, and sexual abuse reports were compared in 127 male and 69 female alcohol‐dependent patients who have a current (36.2%) or lifetime (20.4%) psychiatric disorder or who never had a psychiatric disorder (43.4%). Females reported more emotional and physical abuse than males. Females reported drinking smaller volumes of alcohol but on more days than males. All with current comorbidity, irrespective of gender, reported more days of heavy drinking than other groups. When evaluating drinking status, gender and comorbidity should be considered.
Screening for fetal alcohol syndrome: is it feasible and necessary?
Authors: Burd Larry , Cox Chaun , Fjelstad Kari , McCulloch Kimberly
Published: July 2002

The potential to utilize screening strategies to improve the identification and outcome of persons with fetal alcohol syndrome (FAS) is reviewed. FAS is a condition where screening and surveillance activities would be appropriate. Development of FAS screening and surveillance programs is encouraged because the disorder is expensive. People with FAS have poor outcomes as adults with less than 10% living independently. Several useful tools and models are available. Screening would improve ascertainment and prevalence estimates. Early identification could improve access to services and long term outcome, secondary disabilities and, by extension, excess disability in affected children could be decreased. Lastly, mothers who are at the highest risk to have additional children with FAS could be identified and offered treatment. While both screening and surveillance activities are discussed, the principle focus of this article is a review of the screening process. Two screening tools and several screening methodologies for FAS are available. Since no test will be appropriate in all settings, screening tests need to be selected depending on the setting and population of interest. Screening for FAS should be conducted in a variety of settings and in populations of both high and moderate risk. The results would also provide important data to influence public policy development and resource allocation. Appropriate evaluation of the efficacy, efficiency and effectiveness of FAS screening tools and methodologies would be important before utilization in screening programs.
Cardiovascular and hormonal responses to hyperthermic stress in cocaine addicts after a long period of abstinence
Authors: Vescovi Pier Paolo
Published: July 2002

In order to study the effects of exposure to heat, cardiovascular and hormonal responses were measured in 10 male cocaine addicts (after 4 weeks and 1 year of abstinence) and in 10 normal men. Subjects sat for 30 minutes in a sauna room. Hormonal (ACTH, beta endorphin, (beta EP) met enkephalin (met‐enk), prolactin (PRL), cortisol) and cardiovascular parameters (heart rate, systolic and diastolic blood pressure) were measured before and after heat exposure. Basal levels of ACTH, beta EP, Met‐Enk and cortisol were similar in normal and in cocaine addicts, whereas plasma PRL values were higher in drug abusers after long‐term abstinence. All the examined hormones, with the exception of Met‐Enk, were significantly raised in normal control subjects at the end of sauna. In contrast, no significant hormonal response to hyperthermia was observed in cocaine addicts either after 4 weeks and 1 year of abstinence. No changes were detected in heart rate and blood pressure in each group, indicating that the cardiovascular adaptive responses to hyperthermia in cocaine addicts was unaltered. The results of the present study provide evidence of an impairment of the hormonal response to hyperthermia in cocaine abusers. In conclusion, cocaine abuse produces alteration in the hypothalamic‐pituitary function which persists after a long period of abstinence.
Book and Media Reviews
Authors: COOK CHRIS
Published: July 2002

Books reviewed in this article:

Book reviews in this column will primarily be of titles focusing completely, or in part, on biological aspects of addiction. However, significant titles of general relevance to the addictions field will also be included, even if they are not “biological”, as will titles of general methodological and clinical relevance, even if they are not on “addictions”. Similar considerations will apply to other media (software, audio tapes and CDs, videos, etc). However, specific “addictions” software applications seem to be relatively uncommon and, as these items are rarely reviewed elsewhere, we will endeavour to include reviews of some of the older programmes that are still useful, as well as new titles that appear. I would appreciate suggesti ons of any items suitable for reviews, but especially software and other media of specific relevance to the addictions. Please contact: Professor Chris Cook, Kent Institut e of Medicine and Health Sciences, University of Kent at Canterbury, Kent CT2 7NR, UK. Email: c.c.h.cook@ukc.ac.uk

Substance Abuse: a Comprehensive Textbook, 3rd Edition J. H. LOWINSON, P. RUIZ, R. B. MILLMAN & J. G. LANGROD (Eds)

Women and Substance Abuse–Gender Transparency SALLY J. STEVENS & HARRY K. WEXLER (Eds)

Women, Drug Use and HIV Infection SALLY J. STEVENS, STEPHANIE TORTU & SUSAN L. COYLE

If God Is My Father, How Can He Love Me? CELIA HADDON

Encyclopaedia of Mental Health, Vols 1–3 HOWARD S. FRIEDMAN (Editor‐in‐Chief)

Handbook of Religion and Mental Health H. G. KOENIG (Ed.)

Software
 
StatView v5.0
Fine‐Needle Aspiration Cytology of Soft Tissue Sarcoma: Benefits and Limitations
Authors: Åkerman Måns
Published: July 2002

Purpose. Examine the benefits and limitations of fine‐needle aspiration cytology (FNA) used as the definitive diagnostic method before treatment.

Method. Review of the 25 year experience at a multidisciplinary musculo‐skeletal centre where FNA is the primary diagnostic approach to soft tissue sarcoma in the extremities and trunk wall and the experience of various experts in the field.

Results. FNA has several benefits compared with coarse needle or open surgical biopsy. The most important are rapid preliminary diagnosis, no need for hospitalization and anaesthesia, negligible complications and fear for tumour cell spread. With the collected experience gained during the years a reliable diagnosis of sarcoma is the rule in general and specific‐type diagnoses are possible in many histotypes, especially when the cytologic examination is supplemented with ancillary diagnostics. The most important limitations are inability to hit small deep‐seated sarcoma and some diagnostic pitfalls such as the correct diagnosis of spindle cell neoplasms, variants of benign lipomatous tumours and ‘new soft tissue tumour entities’.

Discussion. Optimal use of FNA calls for certain requirements such as centralization, experience in soft tissue tumour cytology–histopathology, the FNA technique and close co‐operation between the orthopaedic surgeon and cytopathologist.
Glutathione S‐Transferase, Similar to Sigma Class, from Skin Secretions of Xenopus laevis
Authors: Pennelli Alfonso , Ortolano Saida , Miele Rossella , Renda Tindaro , Sacchetta Paolo , Ilio Carmine Di , Simmaco Maurizio
Published: July 2002

Using glutathione affinity chromatography followed by isoelectro‐focusing, we purified from the skin secretion of Xenopus laevis an isoenzyme of glutathione S‐transferase with an apparent subunit molecular mass of 22.5 kDa and an isoelectric point at pH 5.1. Its N‐terminal amino acid sequence was highly similar to that of the sigma class glutathione S‐transferase, which previously was demonstrated to have a glutathione‐dependent prostaglandin D2 synthase activity. Immunohistochemistry analysis revealed that the isoenzyme was located in the cytoplasm of granular gland cells.
Redox Regulation of Lymphocyte Signaling
Authors: Pani Giovanni , Colavitti Renata , Borrello Silvia , Galeotti Tommaso
Published: July 2002

Compelling evidence exists that reactive oxygen species can deliver intracellular signals in mammalian cells, and elicit a broad array of physiological responses according to the cell type, the oxidative burden and the cellular compartment where radicals are generated. When applied to immune cells, these concepts gain a particular relevance, in relation to the plasticity of immune functions and the biological complexity of lymphocyte response to antigens. Here we review some recent and somehow conflicting observations on the involvement of oxygen radicals and redox balance in lymphocyte activation, and propose models for how radical species could contribute to normal and pathological immunity.
Lessons From Sequenced Genomes. Overlapping Genes in Methanococcus jannaschii?
Authors: Szymanski Maciej , Barciszewski Jan
Published: July 2002

This paper describes our finding on overlapping genes in Methanococcus jannaschii genome. We found that one of the open reading frames (ORFs) within the M. jannaschii genome contains the nucleotide sequence of tRNASer, which raises a serious question of the correctness of the initiation codon assignment for that ORF. We suggest that there are two other possible AUG initiation codons downstream from the TTG triplet, which was initially considered as a translation start site. Only one of the AUG triplets is preceded by the Shine‐Dalgarno sequence that seems to be required for binding the ribosome and initiation of translation.
High‐Yield Expression of Fully Bioactive N‐Terminal Parathyroid Hormone Analog in Escherichia coli
Authors: Sung W. L. , Chan B. S. , Luk C. K. , Zahab D. M. , Willick G. E. , Barbier J.‐R. , Isaacs R. , Maclean S. , Ross V. , Morley P. , Whitfield J. F.
Published: July 2002

A fully active analog of human parathyroid hormone (hPTH) has been produced by recombinant expression in Escherichia coli. Initially, a nucleotide sequence encoding hPTH(1‐34)‐Asp‐Pro was ligated to a proinsulin gene in the plasmid pUC8, for the eventual expression of a fusion protein of 137 amino acids. Unexpectedly, the proinsulin gene and 340 bp downstream were deleted by an unknown mechanism during transformation of the E. coli. This resulted in a new plasmid encoding a small (72‐amino acid) fusion product of hPTH(1‐34)‐Asp35‐Pro36‐X, where X is a 36‐residue “arbitrary” downstream sequence of pUC8. The fusion product was efficiently expressed and the hPTH analog, [Asp35]hPTH‐(1‐35), was readily released by acid cleavage, with a yield of 100 mg/L. This analog had an effective concentration for half‐maximal adenylyl cyclase stimulation (EC50) in rat osteosarcoma cells of 14 nM, which was identical to that for hPTH‐(1‐34). In the ovariectomized rat model of osteoporosis, [Asp35]hPTH‐(1‐35) was fully active as a bone anabolic agent.
Regulation of Cardiac Jak‐2 in Animal Models of Insulin Resistance
Authors: Rojas Fernanda Alvarez , Carvalho Carla R. O. , Paez‐Espinosa Veronica , Saad Mario J. A.
Published: July 2002

Insulin induces phosphorylation and activation of JAK2 tyrosine, as well as its association with STAT1 and SHP2 in insulinsensitive tissues of intact rats, thus demonstrating a new pathway in transduction of insulin signals. We investigated this pathway in hearts of rats in three situations of insulin resistance: 72 h of fasting, chronic treatment with dexamethasone, and acute treatment with epinephrine. The acute treatment with epinephrine showed no difference in insulin‐induced JAK2 tyrosine phosphorylation or JAK2/STAT1 and JAK2/SHP2 association in comparison with the control. In fasted rats the JAK2 protein concentration decreased, accompanied by a decrease in the stoichiometry of the phosphorylation to 70%, an increase in association of JAK2/STAT1 to 160%, and a decrease in JAK2/SHP2 association to 85%. In the dexamethasone‐treated group, the JAK2 protein concentrations increased but the stoichiometry of its phosphorylation decreased to 20%, whereas the JAK2/STAT1 and JAK2/SHP2 associations changed by 70% and 170%, respectively. In fasting and dexamethasone‐treated rats, therefore, insulin‐induced JAK2 tyrosine phosphorylation decreases, and the JAK2 protein expression is differentially regulated such that the insulin‐induced JAK2 association with SHP2 and STAT1 shows opposite interactions with the kinase.
Maesaquinone: A Novel Inhibitor of Plant Mitochondrial Respiratory Enzymes That React with Ubiquinone
Authors: Affourtit Charles , Whitehouse David G. , Moore Anthony L.
Published: July 2002

The effect of maesaquinone, 2‐(14‐nonadecenyl)‐3,6‐dihydroxy5‐methyl‐1,4‐benzoquinone, on plant mitochondrial respiration has been investigated. In mitochondria isolated from thermogenic Arum maculatum spadices, this compound inhibits both cytochrome and alternative pathway activities. Kinetic analyses reveal that this inhibition is the result of potent effects of maesaquinone on the alternative oxidase (ID50 <0.3θM) and complex III (ID50 <5θM). Succinate dehydrogenase and external NADH dehydrogenase are also inhibited, albeit to a lesser extent (∼30% at 1 θM). These data suggest that maesaquinone specifically affects the interaction of the respective enzymes with ubiquinone.
Suppression of Mitochondrial Permeability Transition Pore and Induction of Lymphoma P388 Cell Death by Cyclosporin A
Authors: Teplova V. , Evtodienko Yu. , Odinokova I. , Kruglov A. , Kudrjavtsev A.
Published: July 2002

Suppression of the mitochondrial permeability transition pore (PTP) and induction of lymphoma P388 cell death were studied in the presence of cyclosporin A (CsA) and its derivatives. In experiments with permeabilized P388 cells, CsA and its nonimmunosuppressive derivative N‐methyl‐Val‐4‐CsA, but not cyclosporin H (CsH), enhanced Ca2+ accumulation in mitochondria and suppressed PTP opening. Moreover, CsA was able either itself to induce or to enhance a prooxidant‐induced P388 programmed cell death. Blebbing and formation of apoptotic bodies were among the observed CsA effects. N‐Methyl‐Val‐4‐CsA showed similar effects, but CsH had no effect on P388 cell death. These results show that initial‐stage P388 tumour cell death is not related to PTP opening but can be the result of PTP closing with a corresponding increase in the formation of reactive oxygen species.
Molecular Characterization of abLIM, a Novel Actin-binding and Double Zinc Finger Protein
Authors: Roof Dorothy J. , Hayes Annmarie , Adamian Michael , Chishti Athar H. , Li Tiansen
Published: July 2002

Molecules that couple the actin-based cytoskeleton to intracellular signaling pathways are central to the processes of cellular morphogenesis and differentiation. We have characterized a novel protein, the actin-binding LIM (abLIM) protein, which could mediate such interactions between actin filaments and cytoplasmic targets. abLIM protein consists of a COOH-terminal cytoskeletal domain that is fused to an NH2-terminal domain consisting of four double zinc finger motifs. The cytoskeletal domain is ∼50% identical to erythrocyte dematin, an actin-bundling protein of the red cell membrane skeleton, while the zinc finger domains conform to the LIM motif consensus sequence. In vitro expression studies demonstrate that abLIM protein can bind to F-actin through the dematin-like domain. Transcripts corresponding to three distinct isoforms have a widespread tissue distribution. However, a polypeptide corresponding to the full-length isoform is found exclusively in the retina and is enriched in biochemical extracts of retinal rod inner segments. abLIM protein also undergoes extensive phosphorylation in light-adapted retinas in vivo, and its developmental expression in the retina coincides with the elaboration of photoreceptor inner and outer segments. Based on the composite primary structure of abLIM protein, actin-binding capacity, potential regulation via phosphorylation, and isoform expression pattern, we speculate that abLIM may play a general role in bridging the actin-based cytoskeleton with an array of potential LIM protein-binding partners. The developmental time course of abLIM expression in the retina suggests that the retina-specific isoform may have a specialized role in the development or elaboration of photoreceptor inner and outer segments.
Different cis-Regulatory DNA Elements Mediate Developmental Stage- and Tissue-specific Expression of the Human COL2A1 Gene in Transgenic Mice
Authors: Leung Keith K.H. , Ng Ling Jim , Ho Ken K.Y. , Tam Patrick P.L. , Cheah Kathryn S.E.
Published: July 2002

Expression of the type II collagen gene (human COL2A1, mouse Col2a1) heralds the differentiation of chondrocytes. It is also expressed in progenitor cells of some nonchondrogenic tissues during embryogenesis. DNA sequences in the 5′ flanking region and intron 1 are known to control tissue-specific expression in vitro, but the regulation of COL2A1 expression in vivo is not clearly understood. We have tested the regulatory activity of DNA sequences from COL2A1 on the expression of a lacZ reporter gene in transgenic mice. We have found that type II collagen characteristic expression of the transgene requires the enhancer activity of a 309-bp fragment (+2,388 to +2,696) in intron 1 in conjunction with 6.1-kb 5′ sequences. Different regulatory elements were found in the 1.6-kb region (+701 to +2,387) of intron 1 which only needs 90-bp 5′ sequences for tissue-specific expression in different components of the developing cartilaginous skeleton. Distinct positive and negative regulatory elements act together to control tissue-specific transgene expression in the developing midbrain neuroepithelium. Positive elements affecting expression in the midbrain were found in the region from −90 to −1,500 and from +701 to +2,387, whereas negatively acting elements were detected in the regions from −1,500 to −6,100 and +2,388 to +2,855.
Annexin VI-mediated Loss of Spectrin during Coated Pit Budding Is Coupled to Delivery of LDL to Lysosomes
Authors: Kamal Adeela , Ying Yun-shu , Anderson Richard G.W.
Published: July 2002

Previously we reported that annexin VI is required for the budding of clathrin-coated pits from human fibroblast plasma membranes in vitro. Here we show that annexin VI bound to the NH2-terminal 28-kD portion of membrane spectrin is as effective as cytosolic annexin VI in supporting coated pit budding. Annexin VI–dependent budding is accompanied by the loss of ∼50% of the spectrin from the membrane and is blocked by the cysteine protease inhibitor N-acetyl-leucyl-leucyl-norleucinal (ALLN). Incubation of fibroblasts in the presence of ALLN initially blocks the uptake of low density lipoprotein (LDL), but the cells recover after 1 h and internalize LDL with normal kinetics. The LDL internalized under these conditions, however, fails to migrate to the center of the cell and is not degraded. ALLN-treated cells have twice as many coated pits and twofold more membrane clathrin, suggesting that new coated pits have assembled. Annexin VI is not required for the budding of these new coated pits and ALLN does not inhibit. Finally, microinjection of a truncated annexin VI that inhibits budding in vitro has the same effect on LDL internalization as ALLN. These findings suggest that fibroblasts are able to make at least two types of coated pits, one of which requires the annexin VI–dependent activation of a cysteine protease to disconnect the clathrin lattice from the spectrin membrane cytoskeleton during the final stages of budding.
A Role for a Protease in Morphogenic Responses during Yeast Cell Fusion
Authors: Elia Lisa , Marsh Lorraine
Published: July 2002

Cell fusion during yeast mating provides a model for signaling-controlled changes at the cell surface. We identified the AXL1 gene in a screen for genes required for cell fusion in both mating types during mating. AXL1 is a pheromone-inducible gene required for axial bud site selection in haploid yeast and for proteolytic maturation of a-factor. Two other bud site selection genes, RSR1, encoding a small GTPase, and BUD3, were also required for efficient cell fusion. Based on double mutant analysis, AXL1 in a MATα strain acted genetically in the same pathway with FUS2, a fusion-dedicated gene. Electron microscopy of axl1, rsr1, and fus2 prezygotes revealed similar defects in nuclear migration, vesicle accumulation, cell wall degradation, and membrane fusion during cell fusion. The axl1 and rsr1 mutants exhibited defects in pheromone-induced morphogenesis. AXL1 protease function was required in MATα strains for fusion during mating. The ability of the Rsr1p GTPase to cycle was required for efficient cell fusion, as it is for bud site selection. During conjugation, vegetative functions may be redeployed under the control of pheromone signaling for mating purposes. Since Rsr1p has been reported to physically associate with Cdc24p and Bem1p components of the pheromone response pathway, we suggest that the bud site selection genes Rsr1p and Axl1p may act to mediate pheromone control of Fus2p-based fusion events during mating.
Recruitment and the Role of Nuclear Localization in Polyglutamine-mediated Aggregation
Authors: Perez Matthew K. , Paulson Henry L. , Pendse Sagun J. , Saionz Sarah J. , Bonini Nancy M. , Pittman Randall N.
Published: July 2002

The inherited neurodegenerative diseases caused by an expanded glutamine repeat share the pathologic feature of intranuclear aggregates or inclusions (NI). Here in cell-based studies of the spinocerebellar ataxia type-3 disease protein, ataxin-3, we address two issues central to aggregation: the role of polyglutamine in recruiting proteins into NI and the role of nuclear localization in promoting aggregation. We demonstrate that full-length ataxin-3 is readily recruited from the cytoplasm into NI seeded either by a pathologic ataxin-3 fragment or by a second unrelated glutamine-repeat disease protein, ataxin-1. Experiments with green fluorescence protein/polyglutamine fusion proteins show that a glutamine repeat is sufficient to recruit an otherwise irrelevant protein into NI, and studies of human disease tissue and a Drosophila transgenic model provide evidence that specific glutamine-repeat–containing proteins, including TATA-binding protein and Eyes Absent protein, are recruited into NI in vivo. Finally, we show that nuclear localization promotes aggregation: an ataxin-3 fragment containing a nonpathologic repeat of 27 glutamines forms inclusions only when targeted to the nucleus. Our findings establish the importance of the polyglutamine domain in mediating recruitment and suggest that pathogenesis may be linked in part to the sequestering of glutamine-containing cellular proteins. In addition, we demonstrate that the nuclear environment may be critical for seeding polyglutamine aggregates.