bluuebunny/crossref_metadata_2025 huggingface dataset:
Patterns of alcohol consumption in 10 European countries participating in the European Prospective Investigation into Cancer and Nutrition (EPIC) project
Authors: Sieri S , Agudo A , Kesse E , Klipstein-Grobusch K , San-José B , Welch AA , Krogh V , Luben R , Allen N , Overvad K , Tjønneland A , Clavel-Chapelon F , Thiébaut A , Miller AB , Boeing H , Kolyva M , Saieva C , Celentano E , Ocké MC , Peeters PHM , Brustad M , Kumle M , Dorronsoro M , Feito A Fernandez , Mattisson I , Weinehall L , Riboli E , Slimani N
Published: March 2003

bjective:

The aim of this study was to compare the quantities of alcohol and types of alcoholic beverages consumed, and the timing of consumption, in centres participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). These centres, in 10 European countries, are characterised by widely differing drinking habits and frequencies of alcohol-related diseases.

Methods:

We collected a single standardised 24-hour dietary recall per subject from a random sample of the EPIC cohort (36 900 persons initially and 35 955 after exclusion of subjects under 35 and over 74 years of age). This provided detailed information on the distribution of alcohol consumption during the day in relation to main meals, and was used to determine weekly consumption patterns. The crude and adjusted (by age, day of week and season) means of total ethanol consumption and consumption according to type of beverage were stratified by centre and sex.

Results:

Sex was a strong determinant of drinking patterns in all 10 countries. The highest total alcohol consumption was observed in the Spanish centres (San Sebastian, 41.4 g day−1) for men and in Danish centres (Copenhagen, 20.9 g day−1) for women. The lowest total alcohol intake was in the Swedish centres (Umeå, 10.2 g day−1) in men and in Greek women (3.4 g day−1). Among men, the main contributor to total alcohol intake was wine in Mediterranean countries and beer in the Dutch, German, Swedish and Danish centres. In most centres, the main source of alcohol for women was wine except for Murcia (Spain), where it was beer. Alcohol consumption, particularly by women, increased markedly during the weekend in nearly all centres. The German, Dutch, UK (general population) and Danish centres were characterised by the highest percentages of alcohol consumption outside mealtimes.

Conclusions:

The large variation in drinking patterns among the EPIC centres provides an opportunity to better understand the relationship between alcohol and alcohol-related diseases.
Trends in self-reported past alcoholic beverage consumption and ethanol intake from 1950 to 1995 observed in eight European countries participating in the European Investigation into Cancer and Nutrition (EPIC)
Authors: Klipstein-Grobusch K , Slimani N , Krogh V , Keil U , Boeing H , Overvad K , Tjønneland A , Clavel-Chapelon F , Thiébaut A , Linseisen J , Schulze MB , Lagiou P , Papadimitrou A , Saieva C , Veglia F , Bueno-de-Mesquita HB , Peeters PHM , Kumle M , Brustad M , García C Martínez , Barricarte A , Berglund G , Weinehall L , Mulligan A , Allen N , Ferrari P , Riboli E
Published: March 2003

Objective:

To describe the trends of self-reported past consumption of alcoholic beverages and ethanol intake from 1950 to 1995 within the European Prospective Investigation into Cancer and Nutrition (EPIC).

Design:

Data on consumption of beer/cider, wine and liqueur/spirits were obtained retrospectively at age 20, 30 and 40 years to calculate average consumption and ethanol intake for the time periods 1950–1975 (at age 20), 1960–1985 (at age 30) and 1970–1995 (at age 40). Regression analysis was conducted with the time period data to assess trends in past alcoholic beverage consumption and ethanol intake with time.

Setting:

The EPIC project.

Subjects:

In total, 392 064 EPIC participants (275 249 women and 116 815 men) from 21 study centres in eight European countries.

Results:

Generally, increases in beer/cider consumption were observed for most EPIC centres for 1950–1975, 1960–1985 and 1970–1995. Trends in wine consumption differed according to geographical location: downward trends with time were observed for men in southern European EPIC centres, upward trends for those in middle/northern European study centres. For women, similar but less pronounced trends were observed. Because wine consumption was the major contributor to ethanol intake for both men and women in most study centres, time trends for ethanol intake showed a similar geographical pattern to that of wine consumption.

Conclusion:

The different trends in alcoholic beverage consumption and ethanol intake suggest that information depicting lifetime history of ethanol intake should be included in analyses of the relationship between ethanol and chronic diseases, particularly in multi-centre studies such as EPIC.
Diversity of dietary patterns observed in the European Prospective Investigation into Cancer and Nutrition (EPIC) project
Authors: Slimani N , Fahey M , Welch A , Wirfält E , Stripp C , Bergström E , Linseisen J , Schulze MB , Bamia C , Chloptsios Y , Veglia F , Panico S , Bueno-de-Mesquita HB , Ocké MC , Brustad M , Lund E , González CA , Barcos A , Berglund G , Winkvist A , Mulligan A , Appleby P , Overvad K , Tjønneland A , Clavel-Chapelon F , Kesse E , Ferrari P , Van Staveren WA , Riboli E
Published: March 2003

Objective:

To describe the diversity in dietary patterns existing across centres/regions participating in the European Prospective Investigation into Cancer and Nutrition (EPIC).

Design and setting:

Single 24-hour dietary recall measurements were obtained by means of standardised face-to-face interviews using the EPIC-SOFT software. These have been used to present a graphic multi-dimensional comparison of the adjusted mean consumption of 22 food groups.

Subjects:

In total, 35 955 men and women, aged 35–74 years, participating in the EPIC nested calibration study.

Results:

Although wide differences were observed across centres, the countries participating in EPIC are characterised by specific dietary patterns. Overall, Italy and Greece have a dietary pattern characterised by plant foods (except potatoes) and a lower consumption of animal and processed foods, compared with the other EPIC countries. France and particularly Spain have more heterogeneous dietary patterns, with a relatively high consumption of both plant foods and animal products. Apart from characteristics specific to vegetarian groups, the UK ‘health-conscious’ group shares with the UK general population a relatively high consumption of tea, sauces, cakes, soft drinks (women), margarine and butter. In contrast, the diet in the Nordic countries, The Netherlands, Germany and the UK general population is relatively high in potatoes and animal, processed and sweetened/refined foods, with proportions varying across countries/centres. In these countries, consumption of vegetables and fruit is similar to, or below, the overall EPIC means, and is low for legumes and vegetable oils. Overall, dietary patterns were similar for men and women, although there were large gender differences for certain food groups.

Conclusions:

There are considerable differences in food group consumption and dietary patterns among the EPIC study populations. This large heterogeneity should be an advantage when investigating the relationship between diet and cancer and formulating new aetiological hypotheses related to dietary patterns and disease.
Perceptions of ethnic discrimination by minority and majority early adolescents in the Netherlands
Authors: Verkuyten Maykel
Published: April 2003

This research investigates perceptions of discrimination among ethnic majority and minority group early adolescents (aged between 10 and 12 years) living in the multi‐ethnic context of the Netherlands. In two studies (N = 679 and N = 2630), personal and group discrimination was examined in terms of name‐calling and social exclusion, and in relation to ethnic identity and family allocentrism. All early adolescents reported more group than personal discrimination. The personal‐group discrimination discrepancy (PGDD) was found independently of ethnic group, gender, allocentrism, and ethnic identity. Hence, the PGDD seems a more general phenomenon that already exists among early adolescents and across different domains. However, minority group participants perceived far more discrimination overall than majority group early adolescents, and the Turkish participants reported more discrimination than the Moroccan and Surinamese early adolescents. Furthermore, family allocentrism was positively related to perceived discrimination among all ethnic groups in Study 2 and among the Dutch in Study 1. In agreement with ethnic identity development models, strength of ethnic identity was not related to perceived discrimination. Ethnic identity was, however, positively related to allocentrism. In both studies, ethnic minority group participants had higher scores for allocentrism and for ethnic identity than majority group participants. In addition, boys had stronger ethnic identity than girls and ethnic identity was negatively associated with perceived discrimination for the boys but not for the girls. It is concluded that in order to understand early adolescents' perception of discrimination it is necessary to pay attention to basic (cognitive) tendencies that cross ethnic lines, to cultural and status differences between the majority group and ethnic minorities as a category and between ethnic minority groups, and to within‐group differences or individual level variables.
The determinants of parents' beliefs about the intelligence their children: A study from Hong Kong
Authors: Furnham Adrian , Rakow Tim , Mak Twiggy
Published: April 2003

One hundred and ninety‐three Hong Kong parents (mean age 42.2 years) were given a structured interview / questionnaire concerning their own and their children's self‐estimated overall and multiple intelligence. Previous research suggested that males tend to give higher overall “g” estimates to their children and themselves than do females, as well as higher scores on mathematical and spatial intelligence (Furnham, 2001). Further, studies in the West suggest that parents think their children are significantly brighter than they are and that their sons are brighter than their daughters. Estimates were lower than those found in Western populations but, even so, males rated their own mathematical and spatial intelligence higher than did females. Hong Kong Chinese parents did not think their sons were brighter than their daughters. The seven multiple intelligences factored into three clear factors for self and children, and regressions indicated that it was “academic” intelligence (verbal, mathematical, spatial) that was most “g” loaded. The child's age and the self‐rated overall IQ of both the parents were the best predictors of the child's overall estimated IQ. Less than a third of the parents had taken an IQ test or believed they measured IQ very well. Those who were more likely to be better educated, had taken an IQ test, and believed intelligence was inherited were more likely to award themselves higher overall IQ scores. Results are compared with the British studies in the same area.
Exploring self‐esteem of Hong Kong Chinese adults cardiac diseases
Authors: Tam Sing‐Fai , Ng Jenny Yuen Yee
Published: April 2003

Low self‐esteem appears to be both a vulnerability factor for cardiac diseases and a result of cardiac diseases. Thus enhancing self‐esteem might facilitate recovery and/or help to prevent recurrence. Since some evidence suggests that self‐esteem might even serve as an important gauge of cardiac rehabilitation, it is therefore important to know more of the construct of self‐esteem to enable the planning of a more promising rehabilitation process. This paper explores the source and basis of the self‐esteem of Hong Kong Chinese with cardiac diseases. One hundred and fifty‐two adults with cardiac diseases and 146 adults not suffering from any major illness participated in this study. The Adult Source of Self‐Esteem Inventory (ASSEI) (Elovson & Fleming, 1989), open‐ended questions on self‐evaluation, and interviewing were the major procedures used to identify the sources and basis of the self‐esteem of persons with and without cardiac diseases. The subjects' important life aspects were identified through interpreting their responses to open‐ended questions and interviews. Moreover, a structured questionnaire on their subjective perceptions of importance and satisfaction in different life areas was used to identify the relationships between discrete self‐concept variables. Factor analysis of their responses to the 20 ASSEI items revealed four factors, namely, Interpersonal Relationship, Personal Quality, Physical Self, and Personal Achievement. We also examined and compared the means and ranks of the ASSEI items as indicated by the subjects. Content analysis of open‐ended questions further confirmed the self‐esteem domains of persons with cardiac diseases. The discrepancy of ideal‐actual physical abilities was found to be more prominent in the cardiac group. It was also found that family is an important entity to Chinese individuals with cardiac diseases. Implications of the findings to rehabilitation of persons with cardiac diseases were also discussed.
Human values and social identities: A study in two collectivist cultures
Authors: Gouveia Valdiney V. , de Albuquerque Francisco José B. , Clemente Miguel , Espinosa Pablo
Published: April 2003

The current study examines the value correlates of social identities (identification with in‐groups and geospatial units) in two collectivist cultures (Brazil, n = 471; Spain, n = 476). According to previous research, it was expected that subjects who score higher on social values would be more identified with the various traditional in‐groups and with the local geospatial units and that social identities would be predicted by different values among Brazilians and Spaniards. Subjects were undergraduate students (228 males; 719 females), with age ranging from 16 to 55 years (M = 21.9; SD = 4.57), from three states in Brazil (Paraíba, Distrito Federal, and São Paulo) and Spain (Pontevedra, Madrid, and Barcelona). They were administered the Individualism‐CollectivismQuestionnaire, the Basic Value Survey, the In‐group Identification Scale, the Geospatial Identification Scale, and a questionnaire with demographic questions (e.g., gender, age, religious affiliation). Results confirm that importance attributed to social values is correlated with traditional in‐group and local geospatial identification, and that values predicting social identities are different across cultures, especially in the case of values related to geospatial identification. Specifically, among the set of social human values, belonging and tradition were most important to explain social identities. Subjects that assigned more importance to privacy were less identified with traditional ingroups, independently of their national culture. Geospatial identification was based on the values religiosity and belonging for Brazilians, and tradition, social order, honesty, and power for Spaniards. In general, these findings corroborate previous ones, suggesting the existence of a common set of values to explain social identity, based on principally normative values (social order and tradition). Moreover, they indicate the specificity and complexity of the geospatial identification in Spain, where the values of honesty and power contribute to explaining it. In this country, it is possible that the determinants of geospatial identification surpass the social orientation endorsed by people.
The relationship between Turkish children's perceptions of marital conflict and their internalizing and externalizing problems
Authors: Ulu İ. Pinar , Fişiloĝlu Hürol
Published: April 2003

The present study investigated the relationship between children's perceptions of marital conflict and children's internalizing and externalizing problems. Additionally, investigating gender and age differences in children's perceptions and the type of problems they exhibited were the other purposes of the study. The sample consisted of 9‐ to 12‐year‐old, nonclinical children from intact families (N = 232), one of their parents, and teachers. The data were gathered by administering the Child Behavior Checklist for Ages 4–18 and the Teacher's Report Form to adult participants and the Children's Perception of Interparental Conflict Scale and the Children's Depression Inventory to the child participants. Findings indicated that there was a significant relationship between children's perceptions of marital conflict and their internalizing and externalizing problems. More specifically, children's perceptions of conflict properties were associated with their internalizing problems in parents', teachers', and children's reports. Children's perceptions of threat were associated with child‐reported depression. Children's perceptions of self‐blame were associated with child‐reported depression, parent‐reported internalizing and externalizing problems, and teacher‐reported externalizing problems. Furthermore, it was found that there were gender and age differences in children's perceptions of marital conflict and their internalizing and externalizing problems. Findings indicated that boys have higher self‐blame scores and teacher‐reported externalizing problems than girls and that girls have more parent‐ and teacher‐reported internalizing problems than boys. Additionally, it was found that 9‐year‐old children have more teacher‐reported internalizing and externalizing problems than 12‐year‐old children. Also, 9‐year‐old boys have higher parent‐reported externalizing problems than 9‐year‐old girls and 9‐year‐old boys have higher parent‐reported externalizing problems than 12‐year‐old boys.
Intercellular calcium communication regulates platelet aggregation and thrombus growth
Authors: Nesbitt Warwick S. , Giuliano Simon , Kulkarni Suhasini , Dopheide Sacha M. , Harper Ian S. , Jackson Shaun P.
Published: March 2003

The ability of platelets to form stable adhesion contacts with other activated platelets (platelet cohesion or aggregation) at sites of vascular injury is essential for hemostasis and thrombosis. In this study, we have examined the mechanisms regulating cytosolic calcium flux during the development of platelet–platelet adhesion contacts under the influence of flow. An examination of platelet calcium flux during platelet aggregate formation in vitro demonstrated a key role for intercellular calcium communication (ICC) in regulating the recruitment of translocating platelets into developing aggregates. We demonstrate that ICC is primarily mediated by a signaling mechanism operating between integrin αIIbβ3 and the recently cloned ADP purinergic receptor P2Y12. Furthermore, we demonstrate that the efficiency by which calcium signals are propagated within platelet aggregates plays an important role in dictating the rate and extent of thrombus growth.
The Ran GTPase cycle is required for yeast nuclear pore complex assembly
Authors: Ryan Kathryn J. , McCaffery J. Michael , Wente Susan R.
Published: March 2003

Here, we report the first evidence that the Ran GTPase cycle is required for nuclear pore complex (NPC) assembly. Using a genetic approach, factors required for NPC assembly were identified in Saccharomyces cerevisiae. Four mutant complementation groups were characterized that correspond to respective mutations in genes encoding Ran (gsp1), and essential Ran regulatory factors Ran GTPase–activating protein (rna1), Ran guanine nucleotide exchange factor (prp20), and the RanGDP import factor (ntf2). All the mutants showed temperature-dependent mislocalization of green fluorescence protein (GFP)-tagged nucleoporins (nups) and the pore-membrane protein Pom152. A decrease in GFP fluorescence associated with the nuclear envelope was observed along with an increase in the diffuse, cytoplasmic signal with GFP foci. The defects did not affect the stability of existing NPCs, and nup mislocalization was dependent on de novo protein synthesis and continued cell growth. Electron microscopy analysis revealed striking membrane perturbations and the accumulation of vesicles in arrested mutants. Using both biochemical fractionation and immunoelectron microscopy methods, these vesicles were shown to contain nups. We propose a model wherein a Ran-mediated vesicular fusion step is required for NPC assembly into intact nuclear envelopes.
The cytosolic entry of diphtheria toxin catalytic domain requires a host cell cytosolic translocation factor complex
Authors: Ratts Ryan , Zeng Huiyan , Berg Eric A. , Blue Clare , McComb Mark E. , Costello Cathy E. , vanderSpek Johanna C. , Murphy John R.
Published: March 2003

In vitro delivery of the diphtheria toxin catalytic (C) domain from the lumen of purified early endosomes to the external milieu requires the addition of both ATP and a cytosolic translocation factor (CTF) complex. Using the translocation of C-domain ADP-ribosyltransferase activity across the endosomal membrane as an assay, the CTF complex activity was 650–800-fold purified from human T cell and yeast extracts, respectively. The chaperonin heat shock protein (Hsp) 90 and thioredoxin reductase were identified by mass spectrometry sequencing in CTF complexes purified from both human T cell and yeast. Further analysis of the role played by these two proteins with specific inhibitors, both in the in vitro translocation assay and in intact cell toxicity assays, has demonstrated their essential role in the productive delivery of the C-domain from the lumen of early endosomes to the external milieu. These results confirm and extend earlier observations of diphtheria toxin C-domain unfolding and refolding that must occur before and after vesicle membrane translocation. In addition, results presented here demonstrate that thioredoxin reductase activity plays an essential role in the cytosolic release of the C-domain. Because analogous CTF complexes have been partially purified from mammalian and yeast cell extracts, results presented here suggest a common and fundamental mechanism for C-domain translocation across early endosomal membranes.
Spindle assembly and cytokinesis in the absence of chromosomes during Drosophila male meiosis
Authors: Bucciarelli Elisabetta , Giansanti Maria Grazia , Bonaccorsi Silvia , Gatti Maurizio
Published: March 2003

Alarge body of work indicates that chromosomes play a key role in the assembly of both acentrosomal and centrosome-containing spindles. In animal systems, the absence of chromosomes either prevents spindle formation or allows the assembly of a metaphase-like spindle that fails to evolve into an ana-telophase spindle. Here, we show that Drosophila secondary spermatocytes can assemble morphologically normal spindles in the absence of chromosomes. The Drosophila mutants fusolo and solofuso are severely defective in chromosome segregation and produce secondary spermatocytes that are devoid of chromosomes. The centrosomes of these anucleated cells form robust asters that give rise to bipolar spindles that undergo the same ana-telophase morphological transformations that characterize normal spindles. The cells containing chromosome-free spindles are also able to assemble regular cytokinetic structures and cleave normally. In addition, chromosome-free spindles normally accumulate the Aurora B kinase at their midzones. This suggests that the association of Aurora B with chromosomes is not a prerequisite for its accumulation at the central spindle, or for its function during cytokinesis.
uPARAP/Endo180 is essential for cellular uptake of collagen and promotes fibroblast collagen adhesion
Authors: Engelholm Lars H. , List Karin , Netzel-Arnett Sarah , Cukierman Edna , Mitola David J. , Aaronson Hannah , Kjøller Lars , Larsen Jørgen K. , Yamada Kenneth M. , Strickland Dudley K. , Holmbeck Kenn , Danø Keld , Birkedal-Hansen Henning , Behrendt Niels , Bugge Thomas H.
Published: March 2003

The uptake and lysosomal degradation of collagen by fibroblasts constitute a major pathway in the turnover of connective tissue. However, the molecular mechanisms governing this pathway are poorly understood. Here, we show that the urokinase plasminogen activator receptor–associated protein (uPARAP)/Endo180, a novel mesenchymally expressed member of the macrophage mannose receptor family of endocytic receptors, is a key player in this process. Fibroblasts from mice with a targeted deletion in the uPARAP/Endo180 gene displayed a near to complete abrogation of collagen endocytosis. Furthermore, these cells had diminished initial adhesion to a range of different collagens, as well as impaired migration on fibrillar collagen. These studies identify a central function of uPARAP/Endo180 in cellular collagen interactions.
Determinants of [Cl−] in recycling and late endosomes and Golgi complex measured using fluorescent ligands
Authors: Sonawane N.D. , Verkman A.S.
Published: March 2003

Chloride concentration ([Cl−]) was measured in defined organellar compartments using fluorescently labeled transferrin, α2-macroglobulin, and cholera toxin B-subunit conjugated with Cl−-sensitive and -insensitive dyes. In pulse-chase experiments, [Cl−] in Tf-labeled early/recycling endosomes in J774 cells was 20 mM just after internalization, increasing to 41 mM over ∼10 min in parallel to a drop in pH from 6.91 to 6.05. The low [Cl−] just after internalization (compared with 137 mM solution [Cl−]) was prevented by reducing the interior-negative Donnan potential. [Cl−] in α2-macroglobulin–labeled endosomes, which enter a late compartment, increased from 28 to 58 mM at 1–45 min after internalization, whereas pH decreased from 6.85 to 5.20. Cl− accumulation was prevented by bafilomycin but restored by valinomycin. A Cl− channel inhibitor slowed endosomal acidification and Cl− accumulation by ∼2.5-fold. [Cl−] was 49 mM and pH was 6.42 in cholera toxin B subunit–labeled Golgi complex in Vero cells; Golgi compartment Cl− accumulation and acidification were reversed by bafilomycin. Our experiments provide evidence that Cl− is the principal counter ion accompanying endosomal and Golgi compartment acidification, and that an interior-negative Donnan potential is responsible for low endosomal [Cl−] early after internalization. We propose that reduced [Cl−] and volume in early endosomes permits endosomal acidification and [Cl−] accumulation without lysis.
An anillin homologue, Mid2p, acts during fission yeast cytokinesis to organize the septin ring and promote cell separation
Authors: Tasto Joseph J. , Morrell Jennifer L. , Gould Kathleen L.
Published: March 2003

Anillin is a conserved protein required for cell division (Field, C.M., and B.M. Alberts. 1995. J. Cell Biol. 131:165–178; Oegema, K., M.S. Savoian, T.J. Mitchison, and C.M. Field. 2000. J. Cell Biol. 150:539–552). One fission yeast homologue of anillin, Mid1p, is necessary for the proper placement of the division site within the cell (Chang, F., A. Woollard, and P. Nurse. 1996. J. Cell Sci. 109(Pt 1):131–142; Sohrmann, M., C. Fankhauser, C. Brodbeck, and V. Simanis. 1996. Genes Dev. 10:2707–2719). Here, we identify and characterize a second fission yeast anillin homologue, Mid2p, which is not orthologous with Mid1p. Mid2p localizes as a single ring in the middle of the cell after anaphase in a septin- and actin-dependent manner and splits into two rings during septation. Mid2p colocalizes with septins, and mid2Δ cells display disorganized, diffuse septin rings and a cell separation defect similar to septin deletion strains. mid2 gene expression and protein levels fluctuate during the cell cycle in a sep1- and Skp1/Cdc53/F-box (SCF)–dependent manner, respectively, implying that Mid2p activity must be carefully regulated. Overproduction of Mid2p depolarizes cell growth and affects the organization of both the septin and actin cytoskeletons. In the presence of a nondegradable Mid2p fragment, the septin ring is stabilized and cell cycle progression is delayed. These results suggest that Mid2p influences septin ring organization at the site of cell division and its turnover might normally be required to permit septin ring disassembly.
Mid2p stabilizes septin rings during cytokinesis in fission yeast
Authors: Berlin Ana , Paoletti Anne , Chang Fred
Published: March 2003

Septins are filament-forming proteins with a conserved role in cytokinesis. In the fission yeast Schizosaccharomyces pombe, septin rings appear to be involved primarily in cell–cell separation, a late stage in cytokinesis. Here, we identified a protein Mid2p on the basis of its sequence similarity to S. pombe Mid1p, Saccharomyces cerevisiae Bud4p, and Candida albicans Int1p. Like septin mutants, mid2Δ mutants had delays in cell–cell separation. mid2Δ mutants were defective in septin organization but not contractile ring closure or septum formation. In wild-type cells, septins assembled first during mitosis in a single ring and during septation developed into double rings that did not contract. In mid2Δ cells, septins initially assembled in a single ring but during septation appeared in the cleavage furrow, forming a washer or disc structure. FRAP studies showed that septins are stable in wild-type cells but exchange 30-fold more rapidly in mid2Δ cells. Mid2p colocalized with septins and required septins for its localization. A COOH-terminal pleckstrin homology domain of Mid2p was required for its localization and function. No genetic interactions were found between mid2 and the related gene mid1. Thus, these studies identify a new factor responsible for the proper stability and function of septins during cytokinesis.
Caspase cleavage product of BAP31 induces mitochondrial fission through endoplasmic reticulum calcium signals, enhancing cytochrome c release to the cytosol
Authors: Breckenridge David G. , Stojanovic Marina , Marcellus Richard C. , Shore Gordon C.
Published: March 2003

Stimulation of cell surface death receptors activates caspase-8, which targets a limited number of substrates including BAP31, an integral membrane protein of the endoplasmic reticulum (ER). Recently, we reported that a caspase-resistant BAP31 mutant inhibited several features of Fas-induced apoptosis, including the release of cytochrome c (cyt.c) from mitochondria (Nguyen, M., D.G. Breckenridge, A. Ducret, and G.C. Shore. 2000. Mol. Cell. Biol. 20:6731–6740), implicating ER-mitochondria crosstalk in this pathway. Here, we report that the p20 caspase cleavage fragment of BAP31 can direct pro-apoptotic signals between the ER and mitochondria. Adenoviral expression of p20 caused an early release of Ca2+ from the ER, concomitant uptake of Ca2+ into mitochondria, and mitochondrial recruitment of Drp1, a dynamin-related protein that mediates scission of the outer mitochondrial membrane, resulting in dramatic fragmentation and fission of the mitochondrial network. Inhibition of Drp1 or ER-mitochondrial Ca2+ signaling prevented p20-induced fission of mitochondria. p20 strongly sensitized mitochondria to caspase-8–induced cyt.c release, whereas prolonged expression of p20 on its own ultimately induced caspase activation and apoptosis through the mitochondrial apoptosome stress pathway. Therefore, caspase-8 cleavage of BAP31 at the ER stimulates Ca2+-dependent mitochondrial fission, enhancing the release of cyt.c in response to this initiator caspase.
CH-ILKBP regulates cell survival by facilitating the membrane translocation of protein kinase B/Akt
Authors: Fukuda Tomohiko , Guo Lida , Shi Xiaohua , Wu Chuanyue
Published: March 2003

Cell survival depends on proper propagation of protective signals through intracellular signaling intermediates. We report here that calponin homology domain–containing integrin-linked kinase (ILK)–binding protein (CH-ILKBP), a widely expressed adaptor protein localized at plasma membrane-actin junctions, is essential for transmission of survival signals. Cells that are depleted of CH-ILKBP undergo extensive apoptosis despite the presence of cell–extracellular matrix contacts and soluble growth factors. The activating phosphorylation of protein kinase B (PKB/Akt), a key regulator of apoptosis, is impaired in the absence of CH-ILKBP. Importantly, loss of CH-ILKBP prevents the membrane translocation of PKB/Akt. Furthermore, forced membrane targeting of PKB/Akt bypasses the requirement of CH-ILKBP for the activating phosphorylation of PKB/Akt, suggesting that CH-ILKBP is required for the membrane translocation but not the subsequent phosphorylation of PKB/Akt. Finally, we show that loss of CH-ILKBP is also required for the full activation of extracellular signal–regulated kinase (ERK)1/2. However, restoration of the PKB/Akt activation is sufficient for protection of cells from apoptosis induced by the depletion of CH-ILKBP despite the persistent suppression of the ERK1/2 activation. Thus, CH-ILKBP is an important component of the prosurvival signaling pathway functioning primarily by facilitating the membrane translocation of PKB/Akt and consequently the activation of PKB/Akt in response to extracellular survival signals.
To be helped or not helped, that is the question
Authors: Lemichez Emmanuel , Boquet Patrice
Published: March 2003

Diphtheria toxin (DT)**Abbreviations used in this paper: DT, diphtheria toxin; DT-C, DT catalytic fragment; DT-T, DT transmembrane; EF-2, elongation factor 2. is the paradigm of the powerful A-B toxins. These bacterial poisons bind to cells, are endocytosed, and inject their catalytic domain into the cytosol causing the irreversible modification of a key component of the the host cellular machinery. The mechanism by which the hydrophilic enzymatic fragment of DT crosses the endosomal membrane and is released into the cytosol remains controversial. In this issue, Ratts et al. (2003) demonstrate that delivery of the DT catalytic domain from the lumen of purified early endosomes to the external medium requires the addition of a cytosolic translocation factor complex composed in part of Hsp90 and thioredoxin reductase.
Movement in Colorado
Authors: Wells William A.
Published: March 2003

The use of complex and three-dimensional environments, plus the promise of collaborative work by a Migration Consortium, are heating up research into cell migration.
Nonmuscle Myosin Motor of Smooth Muscle
Authors: Löfgren Mia , Ekblad Eva , Morano Ingo , Arner Anders
Published: March 2003

Nonmuscle myosin can generate force and shortening in smooth muscle, as revealed by studies of the urinary bladder from mice lacking smooth muscle myosin heavy chain (SM-MHC) but expressing the nonmuscle myosin heavy chains A and B (NM-MHC A and B; Morano, I., G.X. Chai, L.G. Baltas, V. Lamounier-Zepter, G. Lutsch, M. Kott, H. Haase, and M. Bader. 2000. Nat. Cell Biol. 2:371–375). Intracellular calcium was measured in urinary bladders from SM-MHC–deficient and SM-MHC–expressing mice in relaxed and contracted states. Similar intracellular [Ca2+] transients were observed in the two types of preparations, although the contraction of SM-MHC–deficient bladders was slow and lacked an initial peak in force. The difference in contraction kinetics thus do not reflect differences in calcium handling. Thick filaments were identified with electron microscopy in smooth muscle cells of SM-MHC–deficient bladders, showing that NM-MHC can form filaments in smooth muscle cells. Maximal shortening velocity of maximally activated, skinned smooth muscle preparations from SM-MHC–deficient mice was significantly lower and more sensitive to increased MgADP compared with velocity of SM-MHC–expressing preparations. Active force was significantly lower and less inhibited by increased inorganic phosphate. In conclusion, large differences in nucleotide and phosphate binding exist between smooth and nonmuscle myosins. High ADP binding and low phosphate dependence of nonmuscle myosin would influence both velocity of actin translocation and force generation to promote slow motility and economical force maintenance of the cell.
A Chloride Channel at the Basolateral Membrane of the Distal-convoluted Tubule
Authors: Lourdel Stéphane , Paulais Marc , Marvao Pedro , Nissant Antoine , Teulon Jacques
Published: March 2003

The distal-convoluted tubule (DCT) of the kidney absorbs NaCl mainly via an Na+-Cl− cotransporter located at the apical membrane, and Na+, K+ ATPase at the basolateral side. Cl− transport across the basolateral membrane is thought to be conductive, but the corresponding channels have not yet been characterized. In the present study, we investigated Cl− channels on microdissected mouse DCTs using the patch-clamp technique. A channel of ∼9 pS was found in 50% of cell-attached patches showing anionic selectivity. The NPo in cell-attached patches was not modified when tubules were preincubated in the presence of 10−5 M forskolin, but the channel was inhibited by phorbol ester (10−6 M). In addition, NPo was significantly elevated when the calcium in the pipette was increased from 0 to 5 mM (NPo increased threefold), or pH increased from 6.4 to 8.0 (NPo increased 15-fold). Selectivity experiments conducted on inside-out patches showed that the Na+ to Cl− relative permeability was 0.09, and the anion selectivity sequence Cl− ∼ I−> Br− ∼ NO3− > F−. Intracellular NPPB (10−4 M) and DPC (10−3 M) blocked the channel by 65% and 80%, respectively. The channel was inhibited at acid intracellular pH, but intracellular ATP and PKA had no effect. ClC-K Cl− channels are characterized by their sensitivity to the external calcium and to pH. Since immunohistochemical data indicates that ClC-K2, and perhaps ClC-K1, are present on the DCT basolateral membrane, we suggest that the channel detected in this study may belong to this subfamily of the ClC channel family.
Ion Fluxes in Giant Excised Cardiac Membrane Patches Detected and Quantified with Ion-selective Microelectrodes
Authors: Kang Tong Mook , Markin Vladislav S. , Hilgemann Donald W.
Published: March 2003

We have used ion-selective electrodes (ISEs) to quantify ion fluxes across giant membrane patches by measuring and simulating ion gradients on both membrane sides. Experimental conditions are selected with low concentrations of the ions detected on the membrane side being monitored. For detection from the cytoplasmic (bath) side, the patch pipette is oscillated laterally in front of an ISE. For detection on the extracellular (pipette) side, ISEs are fabricated from flexible quartz capillary tubing (tip diameters, 2–3 microns), and an ISE is positioned carefully within the patch pipette with the tip at a controlled distance from the mouth of the patch pipette. Transport activity is then manipulated by solution changes on the cytoplasmic side. Ion fluxes can be quantified by simulating the ion gradients with appropriate diffusion models. For extracellular (intrapatch pipette) recordings, ion diffusion coefficients can be determined from the time courses of concentration changes. The sensitivity and utility of the methods are demonstrated with cardiac membrane patches by measuring (a) potassium fluxes via ion channels, valinomycin, and Na/K pumps; (b) calcium fluxes mediated by Na/Ca exchangers; (c) sodium fluxes mediated by gramicidin and Na/K pumps; and (d) proton fluxes mediated by an unknown electrogenic mechanism. The potassium flux-to-current ratio for the Na/K pump is approximately twice that determined for potassium channels and valinomycin, as expected for a 3Na/2K pump stoichiometery (i.e., 2K/charge moved). For valinomycin-mediated potassium currents and gramicidin-mediated sodium currents, the ion fluxes calculated from diffusion models are typically 10–15% smaller than expected from the membrane currents. As presently implemented, the ISE methods allow reliable detection of calcium and proton fluxes equivalent to monovalent cation currents <1 pA in magnitude, and they allow detection of sodium and potassium fluxes equivalent to <5 pA currents. The capability to monitor ion fluxes, independent of membrane currents, should facilitate studies of both electrogenic and electroneutral ion–coupled transporters in giant patches.
The Pore Region of the Skeletal Muscle Ryanodine Receptor Is a Primary Locus for Excitation-Contraction Uncoupling in Central Core Disease
Authors: Avila Guillermo , O'Connell Kristen M. S. , Dirksen Robert T.
Published: March 2003

Human central core disease (CCD) is caused by mutations/deletions in the gene that encodes the skeletal muscle ryanodine receptor (RyR1). Previous studies have shown that CCD mutations in the NH2-terminal region of RyR1 lead to the formation of leaky SR Ca2+ release channels when expressed in myotubes derived from RyR1-knockout (dyspedic) mice, whereas a COOH-terminal mutant (I4897T) results in channels that are not leaky to Ca2+ but lack depolarization-induced Ca2+ release (termed excitation-contraction [EC] uncoupling). We show here that store depletion resulting from NH2-terminal (Y523S) and COOH-terminal (Y4795C) leaky CCD mutant release channels is eliminated after incorporation of the I4897T mutation into the channel (Y523S/I4897T and Y4795C/I4897T). In spite of normal SR Ca2+ content, myotubes expressing the double mutants lacked voltage-gated Ca2+ release and thus exhibited an EC uncoupling phenotype similar to that of I4897T-expressing myotubes. We also show that dyspedic myotubes expressing each of seven recently identified CCD mutations located in exon 102 of the RyR1 gene (G4890R, R4892W, I4897T, G4898E, G4898R, A4905V, R4913G) behave as EC-uncoupled release channels. Interestingly, voltage-gated Ca2+ release was nearly abolished (reduced ∼90%) while caffeine-induced Ca2+ release was only marginally reduced in R4892W-expressing myotubes, indicating that this mutation preferentially disrupts voltage-sensor activation of release. These data demonstrate that CCD mutations in exon 102 disrupt release channel permeation to Ca2+ during EC coupling and that this region represents a primary molecular locus for EC uncoupling in CCD.
Petrogenesis of Mesozoic, Peraluminous Granites in the Lamoille Canyon Area, Ruby Mountains, Nevada, USA
Authors: Lee Sang-Yun , Barnes Calvin G. , Snoke Arthur W. , Howard Keith A. , Frost Carol D.
Published: April 2003

Two groups of closely associated, peraluminous, two-mica granitic gneiss were identified in the area. The older, sparsely distributed unit is equigranular (EG) with initial εNd ∼ − 8·8 and initial 87Sr/86Sr ∼0·7098. Its age is uncertain. The younger unit is Late Cretaceous (∼80 Ma), pegmatitic, and sillimanite-bearing (KPG), with εNd from −15·8 to −17·3 and initial 87Sr/86Sr from 0·7157 to 0·7198. The concentrations of Fe, Mg, Na, Ca, Sr, V, Zr, Zn and Hf are higher, and K, Rb and Th are lower in the EG. Major- and trace-element models indicate that the KPG was derived by muscovite dehydration melting (<35 km depth) of Neoproterozoic metapelitic rocks that are widespread in the eastern Great Basin. The models are broadly consistent with anatexis of crust tectonically thickened during the Sevier orogeny; no mantle mass or heat contribution was necessary. As such, this unit represents one crustal end-member of regional Late Cretaceous peraluminous granites. The EG was produced by biotite dehydration melting at greater depths, with garnet stable in the residue. The source of the EG was probably Paleoproterozoic metagraywacke. Because EG magmatism probably pre-dated Late Cretaceous crustal thickening, it required heat input from the mantle or from mantle-derived magma.
A divergent INS protein inCaenorhabditis elegansstructurally resembles human insulin and activates the human insulin receptor
Authors: Hua Qing-xin , Nakagawa Satoe H. , Wilken Jill , Ramos Rowena R. , Jia Wenhua , Bass Joseph , Weiss Michael A.
Published: April 2003

Caenorhabditis eleganscontains a family of putative insulin-like genes proposed to regulate dauer arrest and senescence. These sequences often lack characteristic sequence features of human insulin essential for its folding, structure, and function. Here, we describe the structure and receptor-binding properties of INS-6, a single-chain polypeptide expressed in specific neurons. Despite multiple nonconservative changes in sequence, INS-6 recapitulates an insulin-like fold. Although lacking classical receptor-binding determinants, INS-6 binds to and activates the human insulin receptor. Its activity is greater than that of an analogous single-chain human insulin analog.
The death domain kinase RIP has an essential role in DNA damage-induced NF-κB activation
Authors: Hur Gang Min , Lewis Joseph , Yang Qingfeng , Lin Yong , Nakano Hiroyasu , Nedospasov Sergei , Liu Zheng-gang
Published: April 2003

The transcription factor NF-κB is activated when cells are exposed to genotoxic stress. It has been suggested that DNA damage will trigger a cytoplasmic signaling that leads to the activation of IKK and NF-κB, but the signaling components upstream of IKK have not yet been identified. Here we report that the receptor interacting protein, RIP, is the IKK upstream component, essential for the activation of NF-κB by DNA damage. Also, our findings suggest that this NF-κB activation by DNA damage is not mediated by autocrine or TNF-R1 signaling pathway. In wild-type fibroblasts, DNA damage induced by agents such as adriamycin, campthothecin, and ionizing radiation induces NF-κB activation. We found, however, that DNA damage failed to activate NF-κB in RIP−/− fibroblasts. The induction of IκBα degradation by DNA damage was normal in TNF-R1−/−, TRAF2−/−, TRAF5−/− and FADD−/− fibroblasts or when de novo protein synthesis was blocked. More importantly, the reconstitution of RIP expression in RIP−/− cells restores DNA damage-induced NF-κB activation. We also found that RIP forms a complex with IKK in response to DNA damage. Therefore, our study provides a possible mechanism for the initiation of the cytoplasmic signaling to activate NF-κB in response to DNA damage.
Essential role for caspase 8 in T-cell homeostasis and T-cell-mediated immunity
Authors: Salmena Leonardo , Lemmers Benedicte , Hakem Anne , Matysiak-Zablocki Elzbieta , Murakami Kiichi , Au P.Y. Billie , Berry Donna M. , Tamblyn Laura , Shehabeldin Amro , Migon Eva , Wakeham Andrew , Bouchard Denis , Yeh Wen Chen , McGlade Jane C. , Ohashi Pamela S. , Hakem Razqallah
Published: April 2003

Defects in death receptor-mediated apoptosis have been linked to cancer and autoimmune disease in humans. The in vivo role of caspase 8, a component of this pathway, has eluded analysis in postnatal tissues because of the lack of an appropriate animal model. Targeted disruption ofcaspase 8is lethal in utero. We generated mice with a targetedcaspase 8mutation that is restricted to the T-cell lineage. Despite normal thymocyte development in the absence of caspase 8, we observed a marked decrease in the number of peripheral T-cells and impaired T-cell response ex vivo to activation stimuli.caspase 8ablation protected thymocytes and activated T-cells from CD95 ligand but not anti-CD3-induced apoptosis, or apoptosis activated by agents that are known to act through the mitochondria.caspase 8mutant mice were unable to mount an immune response to viral infection, indicating that caspase 8 deletion in T-cells leads to immunodeficiency. These findings identify an essential, cell-stage-specific role for caspase 8 in T-cell homeostasis and T-cell-mediated immunity. This is consistent with the recent identification of caspase 8 mutations in human immunodeficiency.