bluuebunny/crossref_metadata_2025 huggingface dataset:
Form, frequency and burden of sleep problems in general health care: a report from the WHO Collaborative Study on Psychological Problems in General Health Care
Authors: Üstün TB , Privett M , Lecrubier Y , Weiller E , Simon G , Korten A , Bassett SS , Maier W , Sartorius N ,   Investigators of the WHO International Study on Psychological Problems in General Health Care
Published: July 2002

The WHO Collaborative Study on Psychological Problems in General Health Care examined the frequency, form, course and outcome of psychological problems in general health care settings. A total of 25,916 general health care attenders at 15 sites in 14 countries were screened using the 12-item General Health Questionnaire (GHQ-12). Of those screened, 5,438 were assessed in detail using a Primary Health Care version of the Composite International Diagnostic Interview (CIDI-PHC) in conjunction with the Brief Disability Questionnaire, the Social Disability Schedules, a self rated overall health status form and the 28-item General Health Questionnaire. The analysis has shown that sleep problems were common at all sites with: 26.8% of all patients having some form of sleep problem and 15% of the patients examined had trouble falling or staying asleep. Of those with sleep problems, 51.5% had a well-defined International Classification of Diseases 10th Revision (ICD-10) mental disorder (such as depression, anxiety, somatoform disorders or alcohol problems) and 48.5% of those with sleep problems for at least two weeks or more did not fulfil the criteria for any well defined ICD-10 diagnosis. Persons with sleep problems reported a degree of disability in the performance of their daily activities and social roles even when they had no symptoms of psychological disorders. When such symptoms were present the disability was significantly increased.
The survival motor neuron protein interacts with the transactivator FUSE binding protein from human fetal brain
Authors: Williams Barbara Y , Hamilton Susan L , Sarkar Hemanta K
Published: July 2002

To identify interacting proteins of survival motor neuron (SMN) in neurons, a fetal human brain cDNA library was screened using the yeast two‐hybrid system. One identified group of SMN interacting clones encoded the DNA transactivator FUSE binding protein (FBP). FBP overexpressed in HEK293 cells or endogenously expressed in fetal and adult mouse brain bound specifically in vitro to recombinant SMN protein. Furthermore, an anti‐FBP antibody specifically co‐immunoprecipitated SMN when both proteins were overexpressed in HEK293 cells. These results demonstrate that FBP is a novel interacting partner of SMN and suggests a possible role for SMN in neuronal gene expression.
Crosstalk between estrogen receptor α and the aryl hydrocarbon receptor in breast cancer cells involves unidirectional activation of proteasomes
Authors: Wormke Mark , Stoner Matthew , Saville Brad , Safe Stephen
Published: July 2002

2,3,7,8‐Tetrachlorodibenzo‐p‐dioxin (TCDD) is an environmental toxin that activates the aryl hydrocarbon receptor (AhR) and disrupts multiple endocrine signaling pathways. T47D human breast cancer cells express a functional estrogen receptor α (ERα) and AhR, and treatment of these cells with 17β‐estradiol (E2) or TCDD resulted in a rapid proteasome‐dependent decrease in immunoreactive ERα and AhR proteins (>60–80%), respectively. E2 did not affect the AhR, whereas TCDD induced proteasome‐dependent degradation of both the AhR and ERα in T47D and MCF‐7 human breast cancer cells, and these responses were specifically blocked by proteasome inhibitors. Thus, TCDD‐induced degradation of ERα may contribute to the antiestrogenic activity of AhR agonists and this pathway may be involved in AhR‐mediated disruption of other endocrine responses.
Defective terminal differentiation and hypoplasia of the epidermis in mice lacking the Fgf10 gene
Authors: Suzuki Kentaro , Yamanishi Kiyofumi , Mori Osamu , Kamikawa Mami , Andersen Bogi , Kato Shigeaki , Toyoda Tetsuya , Yamada Gen
Published: July 2002

Here, we characterized the skin and hair phenotype of mice lacking the fibroblast growth factor 10 gene (Fgf10), a newly identified member of the fibroblast growth factor family. Histological examination of Fgf10
−/− newborn mouse skin revealed abnormalities in epidermal morphogenesis. The number of proliferating cells in the basal layer was decreased, the granular layer was hypoplastic and lacked distinctive keratohyaline granules and tonofibrils. The expression of loricrin, a marker of epidermal differentiation, was dramatically reduced. Despite the presence of Fgf10 transcripts in normal hair follicles, abnormalities of hair development were not observed in Fgf10
−/− skin. These data suggest that Fgf10 is required for embryonic epidermal morphogenesis but is not essential for hair follicle development.
Genomic Exploration of the Hemiascomycetous Yeasts: 19. Ascomycetes‐specific genes
Authors: Malpertuy Alain , Tekaia Fredj , Casarégola Serge , Aigle Michel , Artiguenave Francois , Blandin Gaëlle , Bolotin-Fukuhara Monique , Bon Elisabeth , Brottier Philippe , de Montigny Jacky , Durrens Pascal , Gaillardin Claude , Lépingle Andrée , Llorente Bertrand , Neuvéglise Cécile , Ozier-Kalogeropoulos Odile , Potier Serge , Saurin William , Toffano-Nioche Claire , Wésolowski-Louvel Micheline , Wincker Patrick , Weissenbach Jean , Souciet Jean-Luc , Dujon Bernard
Published: July 2002

Comparisons of the 6213 predicted Saccharomyces cerevisiae open reading frame (ORF) products with sequences from organisms of other biological phyla differentiate genes commonly conserved in evolution from ‘maverick’ genes which have no homologue in phyla other than the Ascomycetes. We show that a majority of the ‘maverick’ genes have homologues among other yeast species and thus define a set of 1892 genes that, from sequence comparisons, appear ‘Ascomycetes‐specific’. We estimate, retrospectively, that the S. cerevisiae genome contains 5651 actual protein‐coding genes, 50 of which were identified for the first time in this work, and that the present public databases contain 612 predicted ORFs that are not real genes. Interestingly, the sequences of the ‘Ascomycetes‐specific’ genes tend to diverge more rapidly in evolution than that of other genes. Half of the ‘Ascomycetes‐specific’ genes are functionally characterized in S. cerevisiae, and a few functional categories are over‐represented in them.
Substitution of a conserved aspartate allows cation‐induced polymerization of FtsZ
Authors: Scheffers Dirk-Jan , de Wit Janny G , den Blaauwen Tanneke , Driessen Arnold J.M
Published: July 2002

The prokaryotic tubulin homologue FtsZ polymerizes in vitro in a nucleotide dependent fashion. Here we report that replacement of the strictly conserved Asp212 residue of Escherichia coli FtsZ by a Cys or Asn, but not by a Glu residue results in FtsZ that polymerizes with divalent cations in the absence of added GTP. FtsZ D212C and D212N mutants co‐purify with GTP as bound nucleotide, providing an explanation for the unusual phenotype. We conclude that D212 plays a critical role in the coordination of a metal ion and the nucleotide at the interface of two FtsZ monomers.
Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed‐tagged proteins1
Authors: Lauf Undine , Lopez Patricia , Falk Matthias M
Published: July 2002

A novel, brilliantly red fluorescent protein, DsRed has become available recently opening up a wide variety of experimental opportunities for double labeling and fluorescence resonance electron transfer experiments in combination with green fluorescent protein (GFP). Unlike in the case of GFP, proteins tagged with DsRed were often found to aggregate within the cell. Here we report a simple method that allows rescuing the function of an oligomeric protein tagged with DsRed. We demonstrate the feasibility of this approach on the subunit proteins of an oligomeric membrane channel, gap junction connexins. Additionally, DsRed fluorescence was easily detected 12–16 h post transfection, much earlier than previously reported, and could readily be differentiated from co‐expressed GFP. Thus, this approach can eliminate the major drawbacks of this highly attractive autofluorescent protein.
Activation of protein phosphatase 5 by limited proteolysis or the binding of polyunsaturated fatty acids to the TPR domain
Authors: Chen Mao Xiang , Cohen Patricia T.W
Published: July 2002

Protein phosphatase 5 (PP5) exhibits very low phosphatase activity, which can be stimulated >25‐fold by proteolysis. Since proteolysis cleaves the N‐terminal tetratricopeptide repeat (TPR) domain from the catalytic domain, these results indicate that the TPR domain shields the active site. Polyunsaturated fatty acids, such as arachidonic acid, and lipids containing polyunsaturated fatty acids, such as phosphatidylinositol, stimulate both bacterially expressed human and native rabbit PP5 activity >25‐fold towards casein and myelin basic protein. Phosphatidylinositol binds to the TPR domain, and not to the catalytic domain, indicating that activation by polyunsaturated fatty acids is allosteric and that it may occur by movement of the TPR domain to allow substrate access.
Activation of MMP‐9 by neutrophil elastase in an in vivo model of acute lung injury
Authors: Ferry Gilles , Lonchampt Michel , Pennel Laurence , de Nanteuil Guillaume , Canet Emmanuel , Tucker Gordon C
Published: July 2002

The effect of neutrophil elastase on the functional status of gelatinases was studied in an hamster model developed by intratracheal administration of lipopolysaccharide followed by in situ cell activation with phorbol myristate acetate. This resulted in the production in bronchoalveolar lavage fluids, in addition to the matrix metalloproteinase MMP‐9, of a 75 kDa gelatinase associated with collagenolytic activity. Treatment in vivo with an elastase inhibitor abolished the latter activity. Since, in addition, elastase activates in vitro purified MMP‐9 gelatinase into a similar 75 kDa entity, these data suggest that elastase may be a physiological activator of MMP‐9 in vivo.
Identification of the yeast ARG‐11 gene as a mitochondrial ornithine carrier involved in arginine biosynthesis
Authors: Palmieri Luigi , De Marco Valeria , Iacobazzi Vito , Palmieri Ferdinando , Runswick Michael J , Walker John E
Published: July 2002

The ARG‐11 gene in Saccharomyces cerevisiae encodes a protein with the characteristic features of a family of 35 related membrane proteins that are encoded in the fungal genome. Some of them are known to transport various substrates and products across the inner membranes of mitochondria, but the functions of 29 members of the family are unknown. The yeast ARG‐11 protein has been over‐produced as inclusion bodies in Escherichia coli. It has been solubilized in the presence of sarkosyl, re‐constituted into liposomes and shown to transport ornithine in exchange for protons. Its main physiological role is probably to take ornithine synthesized from glutamate in the mitochondrial matrix to the cytosol where it is converted to arginine.
Asp477 is a determinant of the enantioselectivity in yeast transketolase
Authors: Nilsson Ulrika , Hecquet Laurence , Gefflaut Thierry , Guerard Christine , Schneider Gunter
Published: July 2002

The conserved residue Asp477 in yeast transketolase is located in the substrate channel of the enzyme and forms a hydrogen bond with the C2‐hydroxyl group of the acceptor substrate. The significance of this interaction for the recognition of the preferred acceptor substrates, d‐α‐hydroxyaldehydes was investigated by site‐directed mutagenesis. In the wild‐type enzyme the k
cat/K
M values are by three to four orders of magnitude lower for 2‐deoxyaldoses or substrates with l‐configuration at the C2‐atom. In the Asp477Ala mutant, the k
cat/K
M values for d‐α‐hydroxyaldehydes are decreased by a thousandfold, while the k
cat/K
M values for substrates with l‐configuration or 2‐deoxyaldoses are similar to wild‐type enzyme. These results indicate that Asp477 is involved in determining the enantioselectivity of transketolase.
Female sterility in mice lacking the basigin gene, which encodes a transmembrane glycoprotein belonging to the immunoglobulin superfamily
Authors: Kuno Naohiko , Kadomatsu Kenji , Fan Qi-Wen , Hagihara Masako , Senda Takao , Mizutani Shigehiko , Muramatsu Takashi
Published: July 2002

Basigin (Bsg) is a transmembrane glycoprotein belonging to the immunoglobulin superfamily. Bsg knock‐out mice exhibit infertility of both sexes. Based on limited results, defective implantation has been considered to be the cause of the female infertility. We demonstrate here that disruption of the Bsg gene produces the failure of female reproductive processes including not only implantation but also fertilization. Bsg mRNA expression in cumulus cells and basolateral localization of the Bsg protein in the endometrial epithelium further support the importance of Bsg in these processes.
Tyrosine phosphorylation of the TATA element modulatory factor by the FER nuclear tyrosine kinases
Authors: Schwartz Y. , Ben-Dor I. , Navon A. , Motro B. , Nir U.
Published: July 2002

The FER locus in the mouse encodes two tyrosine kinases, p94ferand p51ferT. While p94feraccumulates in the cytoplasm and nucleus of most mammalian cells the expression of p51ferTis restricted to the nucleus of meiotic primary spermatocytes. The cellular function of the FER kinases is not understood, nor has a substrate for these enzymes been characterized. To identify putative substrates of p94ferand p51ferT, the two enzymes were used as ‘baits’ in the yeast two‐hybrid screening system. cDNAs encoding the mouse TATA element modulatory factor (TMF) were repeatedly isolated in this assay. TMF was previously shown to bind the TATA element in RNA polymerase II promoters and impaired their functioning in a cell free transcription system. Both p94ferand p51ferTphosphorylated the TMF protein in in vitro and in vivo kinase assays. Sequential deletions showed that the carboxy‐terminal region of TMF was essential for phosphorylation. In situ hybridization analysis revealed the preferential accumulation of TMF transcripts in meiotic spermatogenic and oogenic cells. p94ferand p51ferTmay thus modulate the suppressive activity of TMF during cellular growth and in defined differentiation processes.
CYP76C2, an Arabidopsis thaliana cytochrome P450 gene expressed during hypersensitive and developmental cell death
Authors: Godiard Laurence , Sauviac Laurent , Dalbin Nathalie , Liaubet Laurence , Callard Didier , Czernic Pierre , Marco Yves
Published: July 2002

The characterisation of an Arabidopsis thaliana cytochrome P450‐encoding cDNA clone, B72, preferentially expressed during the hypersensitive response (HR) provoked by the bacterial pathogen Pseudomonas syringae pathovar maculicola, is reported. The B72 cDNA clone corresponded to the CYP76C2 gene, which belongs to a small multigene family comprising four genes. HR‐triggering bacteria harbouring different avirulence genes induced the accumulation of transcripts of this P450 gene. CYP76C2 gene expression was moreover associated with various processes leading to cell death such as leaf senescence, ageing of cell cultures, wounding as well as with treatment with the necrotising heavy metal salt, lead nitrate.
Cloning, sequencing and functional expression of a novel human thioredoxin reductase
Authors: Gasdaska Pamela Y. , Berggren Margareta M. , Berry Marla J. , Powis Garth
Published: July 2002

The DNA sequence encoding a novel human thioredoxin reductase has been determined. The protein is predicted to have 524 amino acids including a conserved ‐Cys‐Val‐Asn‐Val‐Gly‐Cys catalytic site and a selenocysteine containing C‐terminal ‐Gly‐Cys‐SeCys‐Gly. The predicted molecular mass is 56.5. The newly identified TR sequence exhibits 54% identity to a previously reported human thioredoxin reductase and 37% identity to human glutathione reductase. Transient transfection of human embryonal kidney cells results in a 5‐fold increase in thioredoxin reductase activity but no increase in glutathione reductase activity.
Identification of the genes responsive to etoposide‐induced apoptosis: application of DNA chip technology
Authors: Wang Yixin , Rea Thomas , Bian Junhui , Gray Steve , Sun Yi
Published: July 2002

DNA chip technology was used in an attempt to identify target genes responsible for apoptosis induced by etoposide, a p53 activating topoisomerase II inhibitor used clinically as an antitumor agent. 62 Individual mRNAs whose mass changed significantly were identified after screening oligonucleotide arrays capable of detecting 6591 unique human mRNA species. 12 (Nine induced and three repressed) of the etoposide‐responsive genes were further studied by Northern analysis and an agreement rate of 92% was reached. Among the 12 genes studied, two (WAF1/p21 and PCNA) are known p53 regulatory genes, two (glutathione peroxidase and S100A2 calcium‐binding protein) appear to be the novel p53 target genes and the others appear to be p53‐independent. Based upon these findings, the signalling pathways that possibly mediate etoposide‐induced apoptosis are proposed.
Strong hydrophobic nature of cysteine residues in proteins
Authors: Nagano Nozomi , Ota Motonori , Nishikawa Ken
Published: July 2002

The differences between disulfide‐bonding cystine (Cys_SS) and free cysteine (Cys_SH) residues were examined by analyzing the statistical distribution of both types of residue in proteins of known structure. Surprisingly, Cys_SH residues display stronger hydrophobicity than Cys_SS residues. A detailed survey of atoms which come into contact with the sulfhydryl group (sulfur atom) of Cys_SH revealed those atoms are essentially the same in number and variety as those of the methyl group of isoleucine, but are quite different to those of the hydroxyl group of serine. Moreover, the relationships among amino acids were also determined using the 3D‐profile table of known protein structures. Cys_SH was located in the hydrophobic cluster, along with residues such as Met, Trp and Tyr, and was clearly separated from Ser and Thr in the polar cluster. These results imply that free cysteines behave as strongly hydrophobic, and not hydrophilic, residues in proteins.
Combination of the novel farnesyltransferase inhibitor RPR130401 and the geranylgeranyltransferase‐1 inhibitor GGTI‐298 disrupts MAP kinase activation and G1‐S transition in Ki‐Ras‐overexpressing transformed adrenocortical cells
Authors: Mazet Jean-Luc , Padieu Martine , Osman Hanan , Maume Gabrielle , Mailliet Patrick , Dereu Norbert , Hamilton Andrew D , Lavelle François , Sebti Saïd M , Maume Bernard F
Published: July 2002

To test the Kirsten‐Ras (Ki‐Ras) alternative prenylation hypothesis in malignant transformation, we used a novel farnesyltransferase inhibitor competitive to farnesyl‐pyrophosphate, RPR130401, and a CaaX peptidomimetic geranylgeranyltransferase‐1 inhibitor GGTI‐298. In Ki‐Ras‐overexpressing transformed adrenocortical cells, RPR130401 at 1–10 μM inhibited very efficiently the [3H]farnesyl but not [3H]geranylgeranyl transfer to Ras. However, proliferation of these cells was only slightly sensitive to RPR130401 (IC50=30 μM). GGTI‐298 inhibited the growth of these cells with an IC50 of 11 μM but cell lysis was observed at 15 μM. The combination of 10 μM RPR130401 and 10 μM GGTI‐298 inhibited efficiently (80%) cell proliferation. These combined inhibitors but not each inhibitor alone blocked the cell cycle in G0/G1 and disrupted MAP kinase activation. Thus, combination of two inhibitors, at non‐cytotoxic concentrations, acting on the farnesyl‐pyrophosphate binding site of the farnesyltransferase and the CaaX binding site of the geranylgeranyltransferase‐1 respectively is an efficient strategy for disrupting Ki‐Ras tumorigenic cell proliferation.
Cloning of bovine RANTES mRNA and its expression and regulation in ovaries in the periovulatory period
Authors: Aust Gabriela , Brylla Elke , Lehmann Irina , Kiessling Silke , Spanel-Borowski Katharina
Published: July 2002

RANTES may be one of the chemoattractants involved in stimulating eosinophils and macrophages to migrate selectively into bovine dominant follicles and into developing corpora lutea. We sequenced a 736 bp fragment of the bovine RANTES mRNA encoding the complete protein and defined the ovarian source of RANTES mRNA. As demonstrated by competitive RT‐PCR, follicle‐derived macrophages showed a 100–1000 times higher RANTES mRNA level compared to unpurified granulosa cells or follicle‐derived fibroblasts. By means of in situ hybridization, RANTES mRNA positive macrophages were located in the former thecal layer of the developing corpora lutea.
Family history of pre‐eclampsia as a predictor for pre‐eclampsia in primigravidas
Authors: Cincotta R.B. , Brennecke S.P.
Published: July 2002

Objective: To assess the clinical utility of knowledge of a family history of pre‐eclampsia as a predictor for the development of pre‐eclampsia in primigravid women. Methods: 368 primigravid women were prospectively recruited from the outpatients department of an obstetric teaching hospital. Details of any family history of pre‐eclampsia were obtained from the women and their subsequent obstetric outcomes were observed. Results: Of 368 primigravid women, 34 (9.2%) developed pre‐eclampsia. Eighteen (4.9%) women of the total group stated that they had a mother (12), sister (five) or both (one) who had had pre‐eclampsia. Of these 18 women, five (27.8%) developed pre‐eclampsia. Of the women who had no family history, 29 (8.3%) developed pre‐eclampsia (relative risk, RR = 3.4; 95% CI, 1.5–7.6; P = 0.018). Four (22.2%) of the women with a positive family history developed severe pre‐eclampsia compared to 18 (5.1%) with a negative family history (RR = 4.3; 95% CI, 1.6‐11.5; P = 0.017). Conclusions: In a primigravida, a family history of pre‐eclampsia is associated with a fourfold increased risk of severe pre‐eclampsia. This clinical history identifies a group who warrant close clinical surveillance during pregnancy and who may be suitable for trials of prophylactic interventions.
Biochemical markers in the synovial fluid of glenohumeral joints from patients with rotator cuff tear
Authors: Yoshihara Yasuo , Hamada Kazutoshi , Nakajima Tomotaka , Fujikawa Kyosuke , Fukuda Hiroaki
Published: July 2002

It is known that rotator cuff tears are sometimes accompanied by joint destruction. Our purpose was to elucidate the pathology with this condition. Thirty‐two synovial fluid (SF) samples aspirated from the glenohumeral joints of patients with rotator cuff tears, including 7 with partial‐thickness and 25 with full‐thickness tears of the rotator cuff (10 massive and 15 isolated supraspinatus tendon (SSp) tears), were examined. Collagenase (MMP‐1), stromelysin 1 (MMP‐3), tissue inhibitor of metalloproteinases‐1 (TIMP‐1) and carboxy‐terminal type II procollagen peptide (pCOL II‐C) were measured in the SF using the respective sandwich enzyme immunoassays. Glycosaminoglycan (GAG) was also quantified with a cationic dye binding method using 1,9‐dimethylmethylene blue. Levels of any molecules except pCOL II‐C in the SF appeared to be higher in full‐thickness tears than those in partial‐thickness tears. Moreover, levels of MMP‐1, MMP‐3 and GAG in the SF were significantly higher in massive tears of the rotator cuff in comparison with those in isolated SSp tears. Such significance was not observed in the levels of TIMP‐1 or pCOL II C in the SF. We examined the relation of those levels with operative findings or clinical parameters from full‐thickness tears, and observed significant correlations of the tear size with the levels of MMP‐1, MMP‐3 and GAG in the SF. Although these marker molecules in SF do not always originate from cartilage, our results may indicate the potential for accelerated cartilage‐degrading activity in the glenohumeral joint in massive tears of the rotator cuff. © 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
Analysis of changes in mRNA levels of myoblast‐ and fibroblast‐derived gene products in healing skeletal muscle using quantitative reverse transcription‐polymerase chain reaction
Authors: Best Thomas M. , Shehadeh Samuel E. , Leverson Glen , Michel James T. , Corr David T. , Aeschlimann Daniel
Published: July 2002

Changes in expression of type III αl‐collagen and myosin II heavy chains were characterized in rabbit skeletal muscle following single stretch injury using quantitative reverse transcription‐polymerase chain reaction. Collagen III expression was highly elevated in the injured leg compared with the control limb both at the myotendinous junction and in the distal muscle belly. While upregulation of collagen III expression at the myotendinous junction was maximal on day 1, collagen III expression in the distal muscle belly was unchanged on day 1 but highly elevated by day 3. Over the initial 7‐day period, there was on average a 94% increase in collagen III expression at the myotendinous junction and a 42% increase in the distal muscle belly. On the other hand, there was little difference, in fact, slightly less expression of myosin II isoforms, in the injured leg compared with the control side. Immunohistochemical analysis of injured muscle showed significant collagen III deposition at the myotendinous junction beginning at day 3 post‐injury and still evident by day 14. Focal deposits of type I and III collagen were first apparent in the distal muscle belly by day 3 and striking by day 7. Taken together, the data suggest the formation of connective tissue scar at the injury site and the absence of significant muscle regeneration following muscle stretch. Furthermore, microinjuries distant to the primary site of injury may result in more general muscle fibrosis and scarring. © 2001 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
Coupling of dopamine receptor subtypes to multiple and diverse G proteins
Authors: Sidhu Anita , Niznik Hyman B.
Published: July 2002

The family of five dopamine receptors subtypes activate cellular effector systems through G proteins. Historically, dopamine receptors were thought to only stimulate or inhibit adenylyl cyclase, by coupling to either Gsα or Giα, respectively. Recent studies in transfected cells, reviewed here, have shown that multiple and highly diverse signaling pathways are activated by specific dopamine receptor subtypes. This multiplicity of signaling responses occurs through selective coupling to distinct G proteins and each of the receptors can interact with more than one G protein. Although some of the multiple coupling of dopamine receptors to different G proteins occurs from within the same family of G proteins, these receptors can also couple to G proteins belonging to different families. Such multiple interactions between receptors and G proteins elicits functionally distinct physiological effects which acts to enhance and subsequently suppress the original receptor response, and to activate apparently distinct signaling pathways. In the brain, where coexpression of functionally distinct receptors in heterogenous cells further adds to the complexity of dopamine signaling, minor alterations in receptor/G protein coupling states during either development or in adults, may underlie the imbalanced signaling seen in dopaminergic‐linked diseases such as schizophrenia, Parkinson's disease and attention deficit hyperactivity disorder.
Brain‐derived neurotrophic factor gene organization and transcription in the zebrafish embryo
Authors: Huynh Gigi , Heinrich Gerhard
Published: July 2002

The gene encoding zebrafish brain‐derived neurotrophic factor (BDNF) was cloned from a PAC genomic DNA library. The entire transcription unit was contained in two independently isolated clones that together encompass 120 kb of genomic DNA. The intron/exon organization of the zebrafish gene was found to be identical to that of the mammalian gene but only one promoter has so far been identified. The associated 5′ exon is 67% identical to exon 1c of the rat BDNF gene. A search of the 5′ flank of the cloned promoter for sequence similarities with known transcription factor binding sites revealed potential AP‐1, CREB, and SP1 binding sites. Fusion constructs containing the cloned promoter and 1.7 kb of 5′ flank and an enhanced green fluorescent protein reporter that becomes membrane‐anchored were injected into 1–8 cell stage embryos. Expression was seen in notochord, muscle, epithelial and endothelial cells of the 1‐day‐old embryo in consonance with the endogenous gene. These results demonstrate that the cloned promoter mediates cell‐specific expression.
Veratridine delays apoptotic neuronal death induced by NGF deprivation through a Na+‐dependent mechanism in cultured rat sympathetic neurons
Authors: Tanaka S. , Koike T.
Published: July 2002

Superior‐cervical ganglion (SCG) cells dissociated from newborn rats depend on nerve growth factor (NGF) for survival. Membrane depolarization with elevated K+ is known to prevent neuronal death following NGF deprivation and/or to promote survival via a Ca2+‐dependent mechanism. Here we have exploited the possibility of whether or not a Na+‐dependent pathway for neuronal survival is present in these cells. Veratridine (ec50=40 nM), a voltage‐dependent Na+ channel activator, significantly delayed the onset of apoptotic cell death in NGF‐deprived SCG neurons that had been cultured for 7 days in the presence of NGF. This effect was blocked completely by Na+ channel blockers including tetrodotoxin (TTX, 1 μM), benzamil (25 μM) and flunarizine (1 μM), but was not attenuated by nimodipine (1 μM), an L‐type Ca2+ channel blocker. The saving effect of veratridine on cultured neurons was observed even in low Ca2+ media (0–1.0 mM), but was completely abolished in a low Na+ medium (38 mM). Sodium‐binding benzofuran isophthalate was employed as a fluorescent probe for monitoring the level of cytoplasmic free Na+, which revealed a sustained increase in its level (12.9 mM, 307% of that of control) in response to veratridine (0.75 μM). The TTX or flunarizine completely blocked veratridine‐induced Na+ influx in these cultured neurons. Moreover, no appreciable increase in intracellular Ca2+ was detected under these conditions. Though Na+ channels were effectual in SCG neurons which were freshly isolated from newborn rats, the Na+‐dependent saving effect of veratridine was not observed in these young neurons. These lines of evidence suggest that the death‐suppressing effect of veratridine on cultured SCG neurons depends on the Na+ influx via voltage‐dependent Na+ channels, and suggests the presence of Na+‐dependent regulatory mechanism(s) in neuronal survival.
Simultaneous measurement of [18F]FDOPA metabolism and cerebral blood flow in newborn piglets
Authors: Brust P. , Bauer R. , Walter B. , Bergmann R. , FÜchtner F. , Vorwieger G. , Steinbach J. , Johannsen B. , Zwiener U.
Published: July 2002

Available information on the dopamine (DA) metabolism of the immature brain is rare. In order to establish a useful animal model we have performed PET experiments in anesthetized neonatal pigs using 6‐[18F]‐fluoro‐L‐DOPA (FDOPA) as tracer. In this study, we have simultaneously determined the cerebral blood flow and the rate constant of FDOPA conversion by the aromatic amino acid decarboxylase, the ultimate enzyme in the synthesis of dopamine. The estimated values of FDOPA decarboxylation in the basal ganglia were similar to values calculated in adult animals and humans. However, in contrast to those studies a significant decarboxylation was also found in the frontal cortex and the cerebellum. HPLC analysis of brain samples also revealed extensive and rapid metabolism of FDOPA in the five investigated brain regions. At 8 min after tracer injection about 80% of FDOPA was already converted to FDA and its metabolites. Surprisingly, a rather high fraction (16−21%) of [18F]‐fluoro‐3‐methoxytyramine was found which may indicate a low storage capacity of vesicular DA at this perinatal stage. It is suggested that the findings are related to the ontogenetic development of the dopaminergic system. The knowledge of the regulation of the DA metabolism in the immature brain may have implications for the understanding of neurodevelopmental effects of perinatal oxygen deprivation.
The combined effects of trkB and trkC mutations on the innervation of the inner ear
Authors: Fritzsch Bernd , Barbacid Mariano , Silos‐santiago Inmaculada
Published: July 2002

Previous research has demonstrated that only the two neurotrophins and their cognate receptors are necessary for the support of the inner ear innervation. However, detailed analyses of patterns of innervation in various combinations of neurotrophin receptor mutants are lacking. We provide here such an analysis of the distribution of afferent and efferent fibers to the ear in various combinations of neurotrophin receptor mutants using the lipophilic tracer DiI. In the vestibular system, trkC+/− heterozygosity aggravates the trkB−/− mutation effect and causes almost complete loss of vestibular neurons. In the cochlea innervation, various mutations are each characterized by specific topological absence of spiral neurons in Rosenthals canal of the cochlea. trkC−/− mutation alone or in combination with trkB+/− heterozygosity causes absence of all basal turn spiral neurons and afferent fibers extend from the middle turn to the basal turn along inner hair cells with little or no contribution to outer hair cells. Both types of basal turn spiral neurons appear to develop and project via radial fibers to inner and, more sparingly, outer hair cells. Simple trkB−/− mutations show a reduction of fibers to outer hair cells in the apex and, less obvious, in the basal turn. Basal turn spiral neurons may be the only neurons present at birth in the cochlea of a trkB−/− mutant mouse combined with trkC+/− heterozygosity. In addition, the trkB−/− mutation combined with trkC+/− heterozygosity has a patchy and variable loss of middle turn spiral neurons in mice of different litters. Comparisons of patterns of innervation of afferent and efferent fibers show a striking similarity of absence of fibers to topologically corresponding areas. For example, in trkC−/− mutants afferents reach the basal turn, spiraling along the cochlea, rather than through radial fibers and efferent fibers follow the same pathway rather than emanating from intraganglionic spiral fibers. The data presented suggest that there are regional specific effects with some bias towards a specific spiral ganglion type : trkC is essential for support of basal turn spiral neurons whereas trkB appears to be more important for middle and apical turn spiral neurons.
Sex and age differences of neurons expressing gaba‐immunoreactivity in the rat bed nucleus of the stria terminalis
Authors: Stefanova Nadya , Bozhilova‐pastirova Anastassia , Ovtscharoff Wladimir
Published: July 2002

Neurons, containing GABA were visualised immunohistochemically in the bed nucleus of the stria terminalis. Young prepubertal (20 days of age) and postpubertal (3 months and 1 year of age) Sprague‐Dawley rats were used. Quantitative studies revealed greater density of GABA‐immunoreactive perikarya in female than in male bed nucleus of the stria terminalis. This difference was not due to distribution in different volumes, since the volumes of the bed nucleus of the stria terminalis in the three ages studied did not differ by gender. Castration of new‐born male rats caused elevation of the density of GABA‐immunoreactive neurons in the bed nucleus of the stria terminalis to female levels on the third month of life. The percentage of nerve cells, expressing detectable amounts of GABA increased with age in the rat bed nucleus of the stria terminalis. The sexual dimorphism of GABA‐immunoreactive neurons in the bed nucleus of the stria terminalis may contribute to the formation of reproductive behavior. The elevation of GABA expression with age might reflect change of the cellular activity in this part of the limbic circuitry.
Are beta‐blockers effective in patients who develop heart failure soon after myocardial infarction? A meta‐regression analysis of randomised trials
Authors: Houghton Timothy , Freemantle Nick , Cleland John G.F.
Published: July 2002

Background:

The great majority of post‐infarction studies of beta‐blockers were conducted in an era when these agents were widely held to be contra‐indicated for the management of heart failure. We now know that beta‐blockers are highly effective for the management of patients with chronic stable heart failure. However, there remains uncertainty about their role in the setting of post‐infarction heart failure and ventricular dysfunction.

Aim:

the primary objective in this paper, was to investigate the extent to which heart failure or evidence of major cardiac dysfunction influenced outcome in previous trials of beta‐blockers in heart failure after myocardial infarction.

Methods:

We assessed the extent to which the inclusion of patients with heart failure or major cardiac dysfunction influenced outcome in randomised trials of long‐term use of beta‐blockade after myocardial infarction. The primary analysis was to assess the extent to which the proportion of patients included in each trial with heart failure influenced the relative odds of all‐cause mortality in the trials. All randomised trials without crossover with treatment lasting more than one month and with 50 or more patients were considered. All those that provided information on the proportion of patients with heart failure or major cardiac dysfunction in the original or subsequent articles were included in the analysis.

Results:

Overall treatment with a beta‐blocker was associated with a 22.6% reduction in the odds of death (95% C1 11‐32.3%). There were very few data on the effects of beta‐blockers after myocardial infarction in patients with documented left ventricular systolic dysfunction. In the analysis that included heart failure as a factor, treatment with a beta‐blocker was associated with a non‐significant interaction with the presence of heart failure. However, because the group including heart failure patients were at higher risk, the absolute benefit of treatment with beta‐blockers appeared greater in this group.

Conclusions:

This analysis suggests that the relative benefit of beta‐blockers on mortality after a myocardial infarction is similar in the presence or absence of heart failure but that the absolute benefit may be greater in the former. However, as current clinical practice has changed radically from the time when the majority of these trials were conducted, further trial evidence would be desirable.
Cotargeting of miR‐126‐3p and miR‐221‐3p inhibits PIK3R2 and PTEN, reducing lung cancer growth and metastasis by blocking AKT and CXCR4 signalling
Authors: Di Paolo Daniela , Pontis Francesca , Moro Massimo , Centonze Giovanni , Bertolini Giulia , Milione Massimo , Mensah Mavis , Segale Miriam , Petraroia Ilaria , Borzi Cristina , Suatoni Paola , Brignole Chiara , Perri Patrizia , Ponzoni Mirco , Pastorino Ugo , Sozzi Gabriella , Fortunato Orazio
Published: June 2021

Lung cancer is the leading cause of cancer‐related death worldwide. Late diagnosis and metastatic dissemination contribute to its low survival rate. Since microRNA (miRNA) deregulation triggers lung carcinogenesis, miRNAs might represent an interesting therapeutic tool for lung cancer management. We identified seven miRNAs, including miR‐126‐3p and miR‐221‐3p, that are deregulated in tumours compared with normal tissues in a series of 38 non‐small‐cell lung cancer patients. A negative correlation between these two miRNAs was associated with poor patient survival. Concomitant miR‐126‐3p replacement and miR‐221‐3p inhibition, but not modulation of either miRNA alone, reduced lung cancer cell viability by inhibiting AKT signalling. PIK3R2 and PTEN were validated as direct targets of miR‐126‐3p and miR‐221‐3p, respectively. Simultaneous miRNA modulation reduced metastatic dissemination of lung cancer cells both in vitro and in vivo through CXCR4 inhibition. Systemic delivery of a combination of miR‐126‐3p mimic and miR‐221‐3p inhibitor encapsulated in lipid nanoparticles reduced lung cancer patient‐derived xenograft growth through blockade of the PIK3R2–AKT pathway. Our findings reveal that cotargeting miR‐126‐3p and miR‐221‐3p to hamper both tumour growth and metastasis could be a new therapeutic approach for lung cancer.
Toddlers' expectations of third‐party punishments and rewards following an act of aggression
Authors: Geraci Alessandra , Surian Luca
Published: June 2021

Rewarding someone who defends the victim of an unjust aggression and punishing someone who chose not to defend her may be very important acts of reciprocation in social life. This study investigates whether 21‐month‐olds have some expectations concerning such punishing and rewarding actions. Infants were shown simple puppet shows and were tested using the violation‐of‐expectation paradigm. In Experiment 1, we found that infants looked longer when they saw a bystander puppet punishing the puppet who defended the victim rather than the puppet who did not defend her. This pattern of looking times was reversed when the punishing action was replaced with a rewarding action (Experiment 2). These findings reveal early‐emerging expectations about punitive and reward motivations in third‐party contexts, and provide some support for theoretical claims about the hardwiring of the human mind for cooperation and prosociality.
Adaptive finite‐time tracking control for parameterized nonlinear systems with full state constraints
Authors: Wu Ziwen , Zhang Tianping
Published: June 2021

In this article, the issue of adaptive finite‐time dynamic surface control (DSC) is discussed for a class of parameterized nonlinear systems with full state constraints. Using the property of logarithmic function, a one‐to‐one nonlinear mapping is constructed to transform a constrained system into an unconstrained system with the same structure. The nonlinear filter is constructed to replace the first‐order linear filter in the traditional DSC, and the demand on the filter time constant is reduced. Based on finite‐time stable theory and using modified DSC, the finite‐time controller is designed via DSC. Theoretical analysis shows that all the signals in the closed‐loop system are semiglobal practical finite‐time stable. Furthermore, none of the states are outside the defined open set. In the end, simulation results are presented to demonstrate the effectiveness of the proposed control schemes with both linear filters and nonlinear filters.
Highly Sensitive Mechanoresponsive Smart Windows Driven by Shear Strain
Authors: Li Jianing , Lu Xuegang , Zhang Yin , Ke Xiaoqin , Wen Xiaoxiang , Cheng Fei , Wei Chaoping , Li Yanlin , Yao Kangkang , Yang Sen
Published: June 2021

Advanced optical materials with rational designs and tunable light transmission have been drawing increasing interest due to their great potential in energy‐efficient buildings and on‐demand optical devices. Mechanoresponsive smart windows (SWs) can modulate light transmittance by mechanical actuation, showing high energy efficiency, low cost, and chemical stability. However, current research mainly focuses on tensile strain‐responsive SWs that typically require a large strain to achieve optical transparency switching‐which causes great inconvenience to practical application and fatigue damage to matrix materials. Herein, a novel shear‐responsive SW with high strain sensitivity is fabricated by vertically fixing a Fe3O4@SiO2 nanochains (NCs) array in an elastic polyacrylamide matrix. The flexible SW exhibits optically transparent with all NCs standing vertically to the SW surface at initial relaxation state, which enables a good shielding effect, with NCs tilting along the shearing direction as the strain applied. Critically, a rather small shear displacement (1.5 mm) applied on the surface of SW gives rise to tunable optical states varying from the transparency state of 65% transmittance to the opaque state of 10%. The as‐prepared SW with novel tuning modulation, high shear strain sensitivity, and optical angle‐dependence holds promising potential in smart windows, optical switches, anti‐voyeurism, and etc.
Hydrogel‐Induced Cell Membrane Disruptions Enable Direct Cytosolic Delivery of Membrane‐Impermeable Cargo
Authors: Van Hoeck Jelter , Van de Vyver Thijs , Harizaj Aranit , Goetgeluk Glenn , Merckx Pieterjan , Liu Jing , Wels Mike , Sauvage Félix , De Keersmaecker Herlinde , Vanhove Christian , de Jong Olivier G. , Vader Pieter , Dewitte Heleen , Vandekerckhove Bart , Braeckmans Kevin , De Smedt Stefaan C. , Raemdonck Koen
Published: June 2021

Intracellular delivery of membrane‐impermeable cargo offers unique opportunities for biological research and the development of cell‐based therapies. Despite the breadth of available intracellular delivery tools, existing protocols are often suboptimal and alternative approaches that merge delivery efficiency with both biocompatibility, as well as applicability, remain highly sought after. Here, a comprehensive platform is presented that exploits the unique property of cationic hydrogel nanoparticles to transiently disrupt the plasma membrane of cells, allowing direct cytosolic delivery of uncomplexed membrane‐impermeable cargo. Using this platform, which is termed Hydrogel‐enabled nanoPoration or HyPore, the delivery of fluorescein isothiocyanate (FITC)–dextran macromolecules in various cancer cell lines and primary bovine corneal epithelial cells is convincingly demonstrated. Of note, HyPore demonstrates efficient FITC‐dextran delivery in primary human T cells, outperforming state‐of‐the‐art electroporation‐mediated delivery. Moreover, the HyPore platform enables cytosolic delivery of functional proteins, including a histone‐binding nanobody as well as the enzymes granzyme A and Cre‐recombinase. Finally, HyPore‐mediated delivery of the MRI contrast agent gadobutrol in primary human T cells significantly improves their T1‐weighted MRI signal intensities compared to electroporation. Taken together, HyPore is proposed as a straightforward, highly versatile, and cost‐effective technique for high‐throughput, ex vivo manipulation of primary cells and cell lines.
Sphingosine 1‐Phosphate Liposomes for Targeted Nitric Oxide Delivery to Mediate Anticancer Effects against Brain Glioma Tumors
Authors: Liu Yang , Wang Xiao , Li Jing , Tang Jian , Li Bin , Zhang Yu , Gu Ning , Yang Fang
Published: June 2021

Specifically targeting glioblastoma multiforme (GBM) blood vessels and actively enhancing the permeability of the brain–blood–tumor barrier (BBTB) are two extremely difficult challenges currently hindering the development of effective therapies against GBM. Herein, a liposome drug delivery system (S1P/JS‐K/Lipo) is described, which delivers the nitric oxide (NO) prodrug JS‐K, O2‐(2,4‐dinitrophenyl) 1‐[(4‐ethoxycarbonyl) piperazin‐1‐yl] diazen‐1‐ium‐1,2‐diolate, to GBM tumors using sphingosine‐1‐phosphate (S1P)‐signaling molecules as active targeting lipid ligands. It is revealed that S1P/JS‐K/Lipo actively penetrates the BBTB, aided by caveolin‐1‐mediated transcytosis, and it is demonstrated that the system specifically interacts with S1P receptors (S1PRs), which are highly expressed on GBM cells. Nondestructive ultrasound imaging in GBM mouse models is also utilized to observe microsized NO bubble production from JS‐K, as catalyzed by the glutathione S‐transferases (GSTs) resident in GBM cells. Given that these NO bubbles strongly promote GBM cell death in vivo, the S1PR‐targeted liposome delivery system—which successfully achieves BBTB penetration and tumor targeted delivery of a complex multicomponent drug regimen—represents a promising approach for targeted therapies against GBM and other carcinomas characterized by elevated S1PR expression.