bluuebunny/crossref_metadata_2025 huggingface dataset:
Diacylglycerol analogues activate second messenger‐operated calcium channels exhibiting TRPC‐like properties in cortical neurons
Authors: Tu Peng , Kunert‐Keil Christiane , Lucke Silke , Brinkmeier Heinrich , Bouron Alexandre
Published: November 2008

The lipid diacylglycerol (DAG) analogue 1‐oleoyl‐2‐acetyl‐sn‐glycerol (OAG) was used to verify the existence of DAG‐sensitive channels in cortical neurons dissociated from E13 mouse embryos. Calcium imaging experiments showed that OAG increased the cytosolic concentration of Ca2+([Ca2+]i) in nearly 35% of the KCl‐responsive cells. These Ca2+responses disappeared in a Ca2+‐free medium supplemented with EGTA. Mn2+quench experiments showed that OAG activated Ca2+‐conducting channels that were also permeant to Ba2+. The OAG‐induced Ca2+responses were unaffected by nifedipine or omega‐conotoxin GVIA (Sigma‐Aldrich, Saint‐Quentin Fallavier, France) but blocked by 1‐[β‐(3‐(4‐Methoxyphenyl)propoxy)‐4‐methoxyphenethyl]‐1H‐imidazole hydrochloride (SKF)‐96365 and Gd3+. Replacing Na+ions withN‐methyl‐d‐glucamine diminished the amplitude of the OAG‐induced Ca2+responses showing that the Ca2+entry was mediated via Na+‐dependent and Na+‐independent mechanisms. Experiments carried out with the fluorescent Na+indicator CoroNa Green showed that OAG elevated [Na+]i. Like OAG, the DAG lipase inhibitor RHC80267 increased [Ca2+]i but not the protein kinase C activator phorbol 12‐myristate 13‐acetate. Moreover, the OAG‐induced Ca2+responses were not regulated by protein kinase C activation or inhibition but they were augmented by flufenamic acid which increases currents through C‐type transient receptor potential protein family (TRPC) 6 channels. In addition, application of hyperforin, a specific activator of TRPC6 channels, elevated [Ca2+]i. Whole‐cell patch‐clamp recordings showed that hyperforin activated non‐selective cation channels. They were blocked by SKF‐96365 but potentiated by flufenamic acid. Altogether, our data show the presence of hyperforin‐ and OAG‐sensitive Ca2+‐permeable channels displaying TRPC6‐like properties. This is the first report revealing the existence of second messenger‐operated channels in cortical neurons.
Invasion phylogeography of the Ponto‐Caspian crustacean Limnomysis benedeni dispersing across Europe
Authors: Audzijonyte Asta , Wittmann Karl J. , Ovcarenko Irina , Väinölä Risto
Published: November 2008

Aim  Limnomysis benedeni Czerniavsky, 1882 is a mysid crustacean native to the Ponto‐Caspian (Black and Caspian Sea) rivers and estuaries, and has recently spread across Europe through intentional and unintentional introductions. We explored the structuring of genetic variation in native and non‐native populations with an aim to trace the sources of the invasions, and to infer whether the spread has occurred through a single or multiple invasion waves.

Location  Native estuaries in the Ponto‐Caspian basin (Volga, Don, Dnieper, Dniester, Danube) and the recently colonized range along the Danube–Rhine river systems and Lithuania.

Methods  A fragment of the mitochondrial COI gene was sequenced to assess genetic affinities and diversity in native and recently established populations.

Results  The genetic diversity in the native regions is organized into several strongly diverged haplotype groups or lineages, partly allopatric, partly sympatric. All these lineages have also spread beyond the native range. Even the recent rapid dispersal across Europe along the Danube–Rhine system towards the North Sea basin involved several lineages from the Danube delta sector. The structuring of genetic diversity among invaded sites suggests multiple invasion events to the Danube–Rhine drainage. This contrasts with data from some other Ponto‐Caspian species, where a single haplotype seems to have occupied most invaded areas. There is no evidence that intentionally stocked reservoirs in the Baltic Sea basin would have contributed to further unintentional spread of L. benedeni.

Main conclusions  Limnomysis benedeni is spreading across Europe using the southern invasion corridor. The invasion most likely involved several waves from differentiated sources in the native Danube delta area.
Patterns and Determinants of International Trade Costs in the Food Industry
Authors: Olper Alessandro , Raimondi Valentina
Published: November 2008

This paper documents patterns in international trade costs in processed foods for a large cross‐section of developing and developed countries, during the 1976–2000 period. A trade costs index is inferred from a micro‐founded gravity equation that incorporates bilateral ‘iceberg’ trade costs. For 2000, the trade costs, expressed as weighted average tariff equivalent, range from 73% for the north to 134% for the south countries. The time patterns show an average reduction of about 13% in the observed period that rises to 26% for the emerging countries. However, the same does not occur for south countries. On ranking the trade costs determinants, we find that, on average, geographical and historical factors seem to dominate those of infrastructure and institutions. However, trade policy emerges as an important determinant of the trade costs between north and emerging countries.
Molecular Mechanisms of Alcoholic Fatty Liver
Authors: Purohit Vishnudutt , Gao Bin , Song Byoung‐Joon
Published: November 2008

Alcoholic fatty liver is a potentially pathologic condition which can progress to steatohepatitis, fibrosis, and cirrhosis if alcohol consumption is continued. Alcohol exposure may induce fatty liver by increasing NADH/NAD+ratio, increasing sterol regulatory element‐binding protein‐1 (SREBP‐1) activity, decreasing peroxisome proliferator‐activated receptor‐α (PPAR‐α) activity, and increasing complement C3 hepatic levels. Alcohol may increase SREBP‐1 activity by decreasing the activities of AMP‐activated protein kinase and sirtuin‐1. Tumor necrosis factor‐α (TNF‐α) produced in response to alcohol exposure may cause fatty liver by up‐regulating SREBP‐1 activity, whereas betaine and pioglitazone may attenuate fatty liver by down‐regulating SREBP‐1 activity. PPAR‐α agonists have potentials to attenuate alcoholic fatty liver. Adiponectin and interleukin‐6 may attenuate alcoholic fatty liver by up‐regulating PPAR‐α and insulin signaling pathways while down‐regulating SREBP‐1 activity and suppressing TNF‐α production. Recent studies show that paracrine activation of hepatic cannabinoid receptor 1 by hepatic stellate cell‐derived endocannabinoids also contributes to the development of alcoholic fatty liver. Furthermore, oxidative modifications and inactivation of the enzymes involved in the mitochondrial and/or peroxisomal β‐oxidation of fatty acids could contribute to fat accumulation in the liver.
Alcohol in Moderation, Cardioprotection, and Neuroprotection: Epidemiological Considerations and Mechanistic Studies
Authors: Collins Michael A. , Neafsey Edward J. , Mukamal Kenneth J. , Gray Mary O. , Parks Dale A. , Das Dipak K. , Korthuis Ronald J.
Published: November 2008

In contrast to many years of important research and clinical attention to the pathological effects of alcohol (ethanol) abuse, the past several decades have seen the publication of a number of peer‐reviewed studies indicating the beneficial effects of light‐moderate, nonbinge consumption of varied alcoholic beverages, as well as experimental demonstrations that moderate alcohol exposure can initiate typically cytoprotective mechanisms. A considerable body of epidemiology associates moderate alcohol consumption with significantly reduced risks of coronary heart disease and, albeit currently a less robust relationship, cerebrovascular (ischemic) stroke. Experimental studies with experimental rodent models and cultures (cardiac myocytes, endothelial cells) indicate that moderate alcohol exposure can promote anti‐inflammatory processes involving adenosine receptors, protein kinase C (PKC), nitric oxide synthase, heat shock proteins, and others which could underlie cardioprotection. Also, brain functional comparisons between older moderate alcohol consumers and nondrinkers have received more recent epidemiological study. In over half of nearly 45 reports since the early 1990s, significantly reduced risks of cognitive loss or dementia in moderate, nonbinge consumers of alcohol (wine, beer, liquor) have been observed, whereas increased risk has been seen only in a few studies. Physiological explanations for the apparent CNS benefits of moderate consumption have invoked alcohol’s cardiovascular and/or hematological effects, but there is also experimental evidence that moderate alcohol levels can exert direct “neuroprotective” actions—pertinent are several studies in vivo and rat brain organotypic cultures, in which antecedent or preconditioning exposure to moderate alcohol neuroprotects against ischemia, endotoxin, β‐amyloid, a toxic protein intimately associated with Alzheimer’s, or gp120, the neuroinflammatory HIV‐1 envelope protein. The alcohol‐dependent neuroprotected state appears linked to activation of signal transduction processes potentially involving reactive oxygen species, several key protein kinases, and increased heat shock proteins. Thus to a certain extent, moderate alcohol exposure appears to trigger analogous mild stress‐associated, anti‐inflammatory mechanisms in the heart, vasculature, and brain that tend to promote cellular survival pathways.
The α1‐Adrenergic Receptor Antagonist, Prazosin, Reduces Alcohol Drinking in Alcohol‐Preferring (P) Rats
Authors: Rasmussen Dennis D. , Alexander Laura L. , Raskind Murray A. , Froehlich Janice C.
Published: November 2008

Background:  Preliminary evidence suggest that noradrenergic signaling may play a role in mediating alcohol drinking behavior in both humans and rats. Accordingly, we tested the hypothesis that blockade of α1‐adrenergic receptors will suppress alcohol drinking in rats selectively bred for alcohol preference (P line).

Methods:  Adult male P rats were given 24‐hour access to food and water and scheduled access to a 15% (v/v) alcohol solution for 2 hours daily. Rats were injected IP with the α1‐adrenergic receptor antagonist, prazosin (0, 0.5, 1.0, 1.5, or 2.0 mg/kg body weight), once a day at 15 minutes prior to onset of the daily 2‐hour 2‐bottle choice, alcohol versus water, access period for 2 consecutive days and then 3 weeks later for 5 consecutive days.

Results:  Prazosin significantly reduced (p < 0.01) alcohol intake during the initial 2 daily administrations, and this reduction of alcohol intake was maintained for 5 consecutive days by daily prazosin treatment in the subsequent more prolonged trial (p < 0.05). The prazosin‐induced reduction of alcohol intake was not dependent upon drug‐induced motor impairment since increases in water drinking (p < 0.05) were exhibited during the 2‐hour access periods during both 2‐ and 5‐day prazosin treatment.

Conclusions:  The results indicate that the noradrenergic system plays a role in mediating alcohol drinking in rats of the P line and suggest that prazosin—a safe, well‐characterized, and well‐tolerated drug—may be an effective pharmacotherapeutic agent for the treatment of alcohol use disorders.
Early Social Isolation in Male Long‐Evans Rats Alters Both Appetitive and Consummatory Behaviors Expressed During Operant Ethanol Self‐Administration
Authors: McCool Brian A. , Chappell Ann M.
Published: November 2008

Background:  Postweaning social isolation in rats produces profound and long‐lasting cognitive and behavioral deficits in adult animals. Importantly, this housing manipulation alters sensitivity to a number of drugs of abuse including ethanol. However, most studies with ethanol have utilized continuous or limited home‐cage access to examine interactions between juvenile social experience and drinking. More recently, social isolation was shown to increased ethanol responding in a “dipper” model of self‐administration (Deehan et al., 2007). In the current study, we utilize a “sipper” operant self‐administration model to distinguish the effects of isolation rearing on ethanol seeking‐ and drinking‐related behaviors.

Methods:  Postweaning juvenile male Long‐Evans rats were placed into 2 housing groups for 6 weeks: one group consisted of individually housed animals; the second group was housed 4 animals per cage. Following the isolation period, anxiety‐like behavior was assessed to confirm the efficacy of the isolation procedure. In some animals, ethanol drinking in the home cage was assessed using a continuous access, 2‐bottle choice paradigm. All animals were then individually housed and trained to lever‐press for a sipper tube containing either an ethanol solution or a sucrose solution.

Results:  Postweaning social isolation increased the expression of anxiety‐like behavior in the elevated plus maze but not the light‐dark box. Ethanol consumption was also increased during continuous home‐cage access with the 2‐bottle choice paradigm. During operant self‐administration, isolation housing increased the response rate and increased ethanol consumption but did not alter responding for or consumption of sucrose. The housing manipulation did not change the total number of lever responses during extinction sessions. Paired‐pulse inhibition deficits that are characteristic of juvenile isolation remained intact after prolonged experience with sucrose self‐administration.

Discussion:  The effects of postweaning social isolation on ethanol drinking in the home cage are also manifest during operant self‐administration. Importantly, these alterations in adult operant self‐administration are ethanol‐specific.
Alcohol‐Induced Tolerance and Physical Dependence in Mice With Ethanol Insensitive α1 GABAAReceptors
Authors: Werner David F. , Swihart Andrew R. , Ferguson Carolyn , Lariviere William R. , Harrison Neil L. , Homanics Gregg E.
Published: November 2008

Background: Although many people consume alcohol (ethanol), it remains unknown why some become addicted. Elucidating the molecular mechanisms of tolerance and physical dependence (withdrawal) may provide insight into alcohol addiction. While the exact molecular mechanisms of ethanol action are unclear, γ‐aminobutyric acid type A receptors (GABAA‐Rs) have been extensively implicated in ethanol action. The α1 GABAA‐R subunit is associated with tolerance and physical dependence, but its exact role remains unknown. In this report, we tested the hypothesis that α1‐GABAA‐Rs mediate in part these effects of ethanol.

Methods: Ethanol‐induced behavioral responses related to tolerance and physical dependence were investigated in knockin (KI) mice that have ethanol‐insensitive α1 GABAA‐Rs and wildtype (WT) controls. Acute functional tolerance (AFT) was assessed using the stationary dowel and loss of righting reflex (LORR) assays. Chronic tolerance was assessed on the LORR, fixed speed rotarod, hypothermia, and radiant tail‐flick assays following 10 consecutive days of ethanol exposure. Withdrawal‐related hyperexcitability was assessed by handling‐induced convulsions following 3 cycles of ethanol vapor exposure/withdrawal. Immunoblots were used to assess α1 protein levels.

Results: Compared with controls, KI mice displayed decreased AFT and chronic tolerance to ethanol‐induced motor ataxia, and also displayed heightened ethanol‐withdrawal hyperexcitability. No differences between WT and KI mice were seen in other ethanol‐induced behavioral measures. Following chronic exposure to ethanol, control mice displayed reductions in α1 protein levels, but KIs did not.

Conclusions: We conclude that α1‐GABAA‐Rs play a role in tolerance to ethanol‐induced motor ataxia and withdrawal‐related hyperexcitability. However, other aspects of behavioral tolerance and physical dependence do not rely on α1‐containing GABAA‐Rs.
In Utero Ethanol Exposure Impairs Defenses Against Experimental Group B Streptococcus in the Term Guinea Pig Lung
Authors: Gauthier Theresa W. , Young Paula A. , Gabelaia Levan , Tang Sonja M. , Ping Xiao‐Du , Harris Frank L. , Brown Lou Ann S.
Published: November 2008

Background:  The effects of fetal alcohol exposure on the risks of neonatal lung injury and infection remain under investigation. The resident alveolar macrophage (AM) is the first line of immune defense against pulmonary infections. In utero ethanol (ETOH) exposure deranges the function of both premature and term guinea pig AM. We hypothesized that fetal ETOH exposure would increase the risk of pulmonary infection in vivo.

Methods:  We developed a novel in vivo model of group B Streptococcus (GBS) pneumonia using our established guinea pig model of fetal ETOH exposure. Timed‐pregnant guinea pigs were pair fed ±ETOH and some were supplemented with the glutathione (GSH) precursor S‐adenosyl‐methionine (SAM‐e). Term pups were given GBS intratracheally while some were pretreated with inhaled GSH prior to the experimental GBS. Neonatal lung and whole blood were evaluated for GBS while isolated AM were evaluated using fluorescent microscopy for GBS phagocytosis.

Results:  Ethanol‐exposed pups demonstrated increased lung infection and sepsis while AM phagocytosis of GBS was deficient compared with control. When SAM‐e was added to the maternal diet containing ETOH, neonatal lung and systemic infection from GBS was attenuated and AM phagocytosis was improved. Inhaled GSH therapy prior to GBS similarly protected the ETOH‐exposed pup from lung and systemic infection.

Conclusions:  In utero ETOH exposure impaired the neonatal lung’s defense against experimental GBS, while maintaining GSH availability protected the ETOH‐exposed lung. This study suggested that fetal alcohol exposure deranges the neonatal lung’s defense against bacterial infection, and support further investigations into the potential therapeutic role for exogenous GSH to augment neonatal AM function.
Characterization of a Functional Polymorphism in the 3′ UTR of SLC6A4 and its Association With Drinking Intensity
Authors: Seneviratne Chamindi , Huang Weihua , Ait‐Daoud Nassima , Li Ming D. , Johnson Bankole A.
Published: November 2008

Background:  The propensity for severe drinking is hypothesized to be regulated by differential expression of serotonin transporter gene (SLC6A4) in the human brain. The SLC6A4 promoter region 5‐HTTLPR has been examined previously as a candidate polymorphic variant associated with severe drinking. In this study, we investigated whether other SLC6A4 single nucleotide polymorphisms (SNPs) are associated with drinking intensity among treatment‐seeking alcoholics and whether these polymorphic variants result in differential SLC6A4 expression levels.

Methods:  We analyzed associations of drinking intensity in 275 (78.5% male) treatment‐seeking alcoholics of Caucasian and Hispanic origin, with 6 SLC6A4 polymorphisms. Next, to examine the functionality of the SNP that showed a significant association with drinking intensity, we transfected the 2 alleles of rs1042173 into HeLa cell cultures and measured serotonin transporter mRNA and protein expression levels by using qRT‐PCR and western blotting techniques.

Results:  One of the 6 polymorphisms we examined, rs1042173 in the 3′ untranslated region (3′‐UTR) of SLC6A4, showed a significant association with drinking intensity. The G allele carriers for rs1042173 were associated with significantly lower drinking intensity (p = 0.0034) compared to T‐allele homozygotes. In HeLa cell cultures, the cells transfected with G allele showed a significantly higher mRNA and protein levels than the T allele‐transfected cells.

Conclusion:  These findings suggest that the allelic variations of rs1042173 affect drinking intensity in alcoholics possibly by altering serotonin transporter expression levels. This provides additional support to the hypothesis that SLC6A4 polymorphisms play an important role in regulating propensity for severe drinking.
Recovery of Activities of Daily Living in Older Adults After Hospitalization for Acute Medical Illness
Authors: Boyd Cynthia M. , Landefeld C. Seth , Counsell Steven R. , Palmer Robert M. , Fortinsky Richard H. , Kresevic Denise , Burant Christopher , Covinsky Kenneth E.
Published: November 2008

OBJECTIVES: To compare functional outcomes in the year after discharge for older adults discharged from the hospital after an acute medical illness with a new or additional disability in their basic self‐care activities of daily living (ADL) (compared with preadmission baseline 2 weeks before admission) with those of older adults discharged with baseline ADL function and identify predictors of failure to recover to baseline function 1 year after discharge.

DESIGN: Observational.

SETTING: Tertiary care hospital, community teaching hospital.

PARTICIPANTS: Older (aged ≥70) patients nonelectively admitted to general medical services (1993–1998).

MEASUREMENTS: Number of ADL disabilities at preadmission baseline and 1, 3, 6, and 12 months after discharge. Outcomes were death, sustained decline in ADL function, and recovery to baseline ADL function at each time point.

RESULTS: By 12 months after discharge, of those discharged with new or additional ADL disability, 41.3% died, 28.6% were alive but had not recovered to baseline function, and 30.1% were at baseline function. Of those discharged at baseline function, 17.8% died, 15.2% were alive but with worse than baseline function, and 67% were at their baseline function (P<.001). Of those discharged with new or additional ADL disability, the presence or absence of recovery by 1 month was associated with long‐term outcomes. Age, cardiovascular disease, dementia, cancer, low albumin, and greater number of dependencies in instrumental ADLs independently predicted failure to recover.

CONCLUSION: For older adults discharged with new or additional disability in ADL after hospitalization for medical illness, prognosis for functional recovery is poor. Rehabilitation interventions of longer duration and timing than current reimbursement allows, caregiver support, and palliative care should be evaluated.
From initial case report to randomized clinical trial through 20 years of research in periodontal therapy
Authors: Nieri Michele , Saletta Daniele , Buti Jacopo , Pagliaro Umberto , Guidi Luisa , Rotundo Roberto , Pini Prato Giovanpaolo
Published: November 2008

Aim: Case reports (CRs) are often the first publication of a new treatment, but randomized clinical trials (RCTs) are needed to confirm the data. The aim of this study was to evaluate how many therapies published as CRs were followed by RCTs of these therapies over a 20‐year period.

Material and Methods: Two researchers conducted a search through international periodontal journals and found the CRs on periodontal treatments published from 1984 to 1986. Subsequent electronic searches made it possible to verify how many of the treatments published as CRs were also investigated through RCTs over the following 20 years.

Results: Thirty‐one different therapies were selected out of the 33 published CRs; 15 (48%) of these 31 treatments were investigated by RCTs over the next 20 years.

Conclusions: As 52% of the CRs were not validated by RCTs, practitioners should view their results with caution.
Small molecule inhibitors of WNT signaling effectively induce apoptosis in acute myeloid leukemia cells
Authors: Minke Katharina Salome , Staib Peter , Puetter Adriane , Gehrke Iris , Gandhirajan Rajesh Kumar , Schlösser Axel , Schmitt Esther Katharina , Hallek Michael , Kreuzer Karl‐Anton
Published: November 2008

In a significant proportion of acute myeloid leukemia (AML) cases the canonical WNT pathway is upregulated and targeting the WNT/LEF1 signaling cascade in AML may be a promising approach to develop new treatments for this entity. Recently two compounds (CGP049090 and PFK115‐584) have been identified, which specifically inhibit complexation of β‐catenin (CTNNB1) and lymphoid enhancer‐binding factor 1 (LEF1) leading to transcriptional inactivation of LEF1 in colon carcinoma cell lines. To evaluate the effect of WNT inhibition utilizing theses compounds with regard to their effectivity in AML we treated the AML cell lines Kasumi‐1 and HL‐60, primary AML blasts and healthy peripheral blood mononuclear cells (PBMCs) with varying concentrations of both substances. Treatment with both compounds for 24 h resulted in a significant killing of AML cell lines and primary AML blasts with 50% effective concentration doses (EC50) within the submicromolar range. PBMCs were not significantly affected as indicated by EC50‐values 100‐fold higher than for AML cells. Cell kill was mediated by apoptosis as indicated by induction of caspases 3 and 7 and cleavage of poly(ADP‐ribose) polymerase (PARP) upon treatment. Furthermore, we could show that both compounds substantially decrease expression of CTNNB1/LEF1 target genes c‐myc, cyclin D1 and survivin, proofing the specificity of the substances. This was shown in both, AML cell lines and most of the tested primary samples. Our data demonstrate that targeting this pathway seems to be an innovative approach in the treatment of AML.
Transcellular migration of neutrophils is a quantitatively significant pathway across dermal microvascular endothelial cells
Authors: Marmon Shana , Cammer Michael , Raine Cedric S. , Lisanti Michael P.
Published: November 2008

Abstract:  Neutrophil extravasation is central to inflammatory skin diseases like psoriasis and atopic dermatitis. In vivo, neutrophils have been shown to migrate through cell‐to‐cell junctions (paracellular pathway) or directly through the body of the endothelial cell (transcellular pathway). In vitro, however, neutrophil migration is a largely paracellular process where cells preferentially cross at tricellular corners devoid of tight junctions. To approximate the type of cells encountered by extravasating neutrophils in vivo, we developed a neutrophil‐migration assay using primary human dermal microvascular endothelial cells. We show here that a large proportion of migrating neutrophils traverse a monolayer of microvascular endothelium using a purely transcellular pathway. In addition, we demonstrate that F‐actin is rearranged similarly in neutrophils undergoing diapedesis along either route. This in vitro model closely simulates the physiological process of neutrophil extravasation in vivo and can be further utilized to evaluate the relative contribution of distinct migratory pathways to the pathophysiology of inflammatory skin disease.
STAM Adaptor Proteins Interact with COPII Complexes and Function in ER‐to‐Golgi Trafficking
Authors: Rismanchi Neggy , Puertollano Rosa , Blackstone Craig
Published: November 2008

Signal‐transducing adaptor molecules (STAMs) are involved in growth factor and cytokine signaling as well as receptor degradation, and they form complexes with a number of endocytic proteins, including Hrs and Eps15. In this study, we demonstrate that STAM proteins also localize prominently to early exocytic compartments and profoundly regulate Golgi morphology. Upon STAM overexpression in cells, the Golgi apparatus becomes extensively fragmented and dispersed, but when STAMs are depleted, the Golgi becomes highly condensed. Under both scenarios, vesicular stomatitis virus G protein–green fluorescent protein trafficking to the plasma membrane is markedly inhibited, and recovery of Golgi morphology after Brefeldin A treatment is substantially impaired in STAM‐depleted cells. Furthermore, STAM proteins interact with coat protein II (COPII) proteins, probably at endoplasmic reticulum (ER) exit sites, and Sar1 activity is required to maintain the localization of STAMs at discrete sites. Thus, in addition to their roles in signaling and endocytosis, STAMs function prominently in ER‐to‐Golgi trafficking, most likely through direct interactions with the COPII complex.
Concentration of GPI‐Anchored Proteins upon ER Exit in Yeast
Authors: Castillon Guillaume A. , Watanabe Reika , Taylor Marcia , Schwabe Tatjana M. E. , Riezman Howard
Published: November 2008

Previous biochemical work has revealed two parallel routes of exit from the endoplasmic reticulum (ER) in the yeast Saccharomyces cerevisiae, one seemingly specific for glycosyl‐phosphatidylinositol (GPI)‐anchored proteins. Using the coat protein II (COPII) mutant sec31‐1, we visualized ER exit sites (ERES) and identified three distinct ERES populations in vivo. One contains glycosylated pro‐α‐factor, the second contains the GPI‐anchored proteins Cwp2p, Ccw14p and Tos6p and the third is enriched with the hexose transporter, Hxt1p. Concentration of GPI‐anchored proteins prior to budding requires anchor remodeling, and Hxt1p incorporation into ERES requires the COPII components Sec12p and Sec16p. Additionally, we have found that GPI‐anchored protein ER exit is controlled by the p24 family member Emp24p, whereas ER export of most transmembrane proteins requires the Cornichon homologue Erv14p.
A Single Method for Cryofixation and Correlative Light, Electron Microscopy and Tomography of Zebrafish Embryos
Authors: Nixon Susan J. , Webb Richard I. , Floetenmeyer Matthias , Schieber Nicole , Lo Harriet P. , Parton Robert G.
Published: November 2008

The zebrafish is a powerful vertebrate system for cell and developmental studies. In this study, we have optimized methods for fast freezing and processing of zebrafish embryos for electron microscopy (EM). We show that in the absence of primary chemical fixation, excellent ultrastructure, preservation of green fluorescent protein (GFP) fluorescence, immunogold labelling and electron tomography can be obtained using a single technique involving high‐pressure freezing and embedding in Lowicryl resins at low temperature. As well as being an important new tool for zebrafish research, the maintenance of GFP fluorescence after fast freezing, freeze substitution and resin embedding will be of general use for correlative light and EM of biological samples.
Export of PfSBP1 to the Plasmodium falciparum Maurer’s Clefts
Authors: Saridaki Theodora , Fröhlich Kathrin S , Braun‐Breton Catherine , Lanzer Michael
Published: November 2008

The human malaria parasite Plasmodium falciparum exports determinants of virulence and pathology to destinations within the host erythrocyte, including the erythrocyte cytoplasm, plasma membrane and membrane profiles of parasite origin termed Maurer’s clefts. Most of the exported proteins contain a conserved pentameric motif termed plasmodial export element (PEXEL)/vacuolar transfer signal (VTS) that functions as a cleavable sorting signal permitting export to the host erythrocyte. However, there are some exported proteins, such as the skeleton‐binding protein 1 (PfSBP1) that lack the PEXEL/VTS motif and that are not N‐terminally processed, suggesting the presence of alternative sorting signals and/or mechanisms. In this study, we have investigated trafficking of PfSBP1 to the Maurer’s clefts. Our data show that the transmembrane domain of PfSBP1 functions as an internal signal sequence for entry into the parasite’s secretory pathway and for transport to the parasite plasma membrane. Trafficking beyond the parasite’s plasma membrane required additional N‐terminal domains, which are characterized by a high negative net charge. Biochemical data indicate that these domains affect the solubility and extraction profile, the orientation of the protein within the membrane and the subcellular localization. Our findings suggest new principles of protein export in P. falciparum‐infected erythrocytes.
Developmental and activity‐dependent genomic occupancy profiles of CREB in monkey area V1
Authors: Lalonde J. , Lachance P. E. D. , Chaudhuri A.
Published: November 2008

The mammalian neocortex displays significant plastic rearrangement in response to altered sensory input, especially during early postnatal development. It is believed that cyclic AMP‐response element‐binding (CREB) plays an important role in orchestrating the molecular events that guide neuroplastic change, although the details of its genomic targets during normal postnatal development or in response to sensory deprivation remain unknown. Here, we performed CREB chromatin immunoprecipitation (ChIP) from monkey area V1 tissue and hybridized enriched DNA fragments to promoter microarrays (ChIP chip analysis). Our goal was to determine and categorize the CREB regulon in monkey area V1 at two distinct developmental stages (peak of critical period vs. adulthood) and after 5 days of monocular enucleation (ME) at both ages. Classification of enriched candidates showed that the majority of isolated promoter loci (n = 795) were common to all four conditions. A particularly interesting group of candidates (n = 192) was specific to samples derived from enucleated infant area V1. Gene ontology analysis of CREB targets during early postnatal development showed a subgroup of genes implicated in cytoskeleton‐based structural modification. Analysis of messenger RNA expression (quantitative real‐time–polymerase chain reaction) of candidate genes showed striking differences in expression profiles between infant and adult area V1 after ME. Our study represents the first extensive genomic analysis of CREB DNA occupancy in monkey neocortex and provides new insight into the multifaceted transcriptional role of CREB in guiding neuroplastic change.
Social approach in genetically engineered mouse lines relevant to autism
Authors: Moy S. S. , Nadler J. J. , Young N. B. , Nonneman R. J. , Grossman A. W. , Murphy D. L. , D’Ercole A. J. , Crawley J. N. , Magnuson T. R. , Lauder J. M.
Published: November 2008

Profound impairment in social interaction is a core symptom of autism, a severe neurodevelopmental disorder. Deficits can include a lack of interest in social contact and low levels of approach and proximity to other children. In this study, a three‐chambered choice task was used to evaluate sociability and social novelty preference in five lines of mice with mutations in genes implicated in autism spectrum disorders. Fmr1tm1Cgr/Y(Fmr1−/y) mice represent a model for fragile X, a mental retardation syndrome that is partially comorbid with autism. We tested Fmr1−/ymice on two genetic backgrounds, C57BL/6J and FVB/N‐129/OlaHsd (FVB/129). Targeted disruption of Fmr1 resulted in low sociability on one measure, but only when the mutation was expressed on FVB/129. Autism has been associated with altered serotonin levels and polymorphisms in SLC6A4 (SERT), the serotonin transporter gene. Male mice with targeted disruption of Slc6a4 displayed significantly less sociability than wild‐type controls. Mice with conditional overexpression of Igf‐1 (insulin‐like growth factor‐1) offered a model for brain overgrowth associated with autism. Igf‐1 transgenic mice engaged in levels of social approach similar to wild‐type controls. Targeted disruption in other genes of interest, En2 (engrailed‐2) and Dhcr7, was carried on genetic backgrounds that showed low levels of exploration in the choice task, precluding meaningful interpretations of social behavior scores. Overall, results show that loss of Fmr1 or Slc6a4 gene function can lead to deficits in sociability. Findings from the fragile X model suggest that the FVB/129 background confers enhanced susceptibility to consequences of Fmr1 mutation on social approach.
Relatively strong automatic appetitive action‐tendencies in male carriers of the OPRM1 G‐allele
Authors: Wiers R. W. , Rinck M. , Dictus M. , Van Den Wildenberg E.
Published: November 2008

This study investigated whether automatic approach action tendencies for alcohol‐related stimuli were associated with variation in the mu‐opioid receptor gene (OPRM1), previously related to rewarding effects of alcohol and craving. An adapted approach avoidance task was used, in which participants pulled or pushed a joystick in reaction to the format of a picture shown on the computer screen (e.g. pull landscape pictures and push portrait pictures). Picture size on the screen changed upon joystick movement, so that upon a pull movement picture size increased (creating a sense of approach) and upon a push movement picture size decreased (avoidance). Participants reacted to four categories of pictures: alcohol‐related, other appetitive, general positive and general negative. The sample consisted of 84 heavy drinking young men without a g‐allele in the A118G (or A355G) single nucleotide polymorphism of the OPRM1 gene and 24 heavy drinking young men with at least one g‐allele. Heavy drinking carriers of a g‐allele showed relatively strong automatic approach tendencies for alcohol (approach bias). Unexpectedly, they also showed an approach bias for other appetitive stimuli. No approach bias was found for general positive or negative stimuli. These results suggest that automatic approach tendencies in response to appetitive stimuli could play a role in the etiology of addictive behaviors and related disorders. Further research is needed to investigate the specificity of this approach bias and possible gender differences.
Shared aetiology of risky sexual behaviour and adolescent misconduct: genetic and environmental influences
Authors: Verweij K. J. H. , Zietsch B. P. , Bailey J. M. , Martin N. G.
Published: November 2008

Risky sexual behaviour (RSB) is a major risk factor for serious diseases as well as unplanned pregnancy. It is not known if RSB has a genetic basis or if it is only influenced by social and cultural conditions. Adolescent conduct disorder has previously been linked to RSB and has been found to be influenced by both genetic and environmental factors. In this study, we look at normal variation in a broad measure of RSB and in retrospectively reported adolescent misconduct in a large community sample of twins (n = 4904) to partition the variance and covariance between the traits into genetic and environmental components. We found that RSB is influenced to the same extent by genes, shared environment and unshared environment. Adolescent misconduct is moderately influenced by genetic factors and only modestly by shared environmental factors. Moreover, RSB is associated with adolescent misconduct (r = 0.5), primarily because of genetic correlation between the variables. The implications of our findings as well as possible sex differences are discussed.
Beowulf and Poststructuralist Theory
Authors: Sharma Manish
Published: November 2008

In this essay, I examine first the emergence of poststructuralist theory in France in the 1960s and discuss the critical and philosophical innovations of the major poststructuralists. Secondly, I investigate the reception of poststructuralist theory by literary critics in general and Anglo‐Saxonists in particular. Thirdly, I review and summarize readings of ‘Beowulf’ informed by poststructuralist theory. Finally, I suggest ways in which poststructuralist thought can continue to inform productively the study of Anglo‐Saxon literary culture.
BeyondDeronda?: Victorian Studies and Jewish Chronicles
Authors: Gracombe Sarah
Published: November 2008

What accounts for the surprising fact that fictional Jews fill the pages of Victorian literature but actual Jews constituted only a sliver of England's population? This article introduces readers to recent scholarship that attempts to answer this question, charting the diverse ways in which Jews were portrayed in the Victorian cultural imaginary. Much of this scholarship pivots around one fundamental contention: that the most significant thing about representations of nineteenth‐century Jewishness is what they have to teach us about nineteenth‐century Englishness. Depicting Jews became a crucial method for Victorians to test some of their guiding assumptions and underlying anxieties about their nationality and its relationship to the categories of race, religion, culture, and territory—categories contested then as now.

After a brief survey of Jewishness's rise in literary studies, I discuss current trends in research treating Jewishness and Englishness; such research is now occurring within the fields of both Victorian Studies and Jewish Studies, productively contesting the disciplinary and geographic borders of each. Particular attention is paid to the engagement of new scholarship with issues of gender, principally representations of and by Jewish women, and genre, including the novel's fraught relationship to liberalism. I then explore the way analyses of Victorian Jewishness are increasingly going global, looking to British imperial and cultural involvement in America, Europe, and the Middle East. These moves have in turn provoked a reassessment of both Orientalism and Englishness as a cultural identity. Above all, this article argues for the value recent scholarship offers by continuing to inform and challenge our understanding not only of Jewishness but the many facets of Victorian culture with which it intersected.
Mapping the Female Self through the Canadian Landscape: Short Stories by Canadian Women Writing in English
Authors: Jenainati Cathia
Published: November 2008

This essay offers a chronological analysis of the progress of the short story in Canada as narrated by women writers who specifically examined the problematic relationship of the individual with the Canadian landscape. It demonstrates that whilst early narratives portrayed settler women as colonizers and tamers of the wilderness, narratives of the first half of the twentieth century chart the complexities – ethnic, political, historical and cultural – of establishing a cartography of the self. The chosen stories cover the range of recorded female experiences from 1845 to 1945.
The Novel (in Theory)
Authors: Kurnick David
Published: November 2008

In a moment in which many critics are speaking of a new formalism, the recent appearance of three major anthologies of new and classic novel criticism bespeaks an unprecedented upsurge of interest in once‐unfashionable questions of genre. And though these new books leave it unclear whether the novel is in fact a genre, they show that novel theory is: a genre defined at its core by an orientation toward totality. This essay examines the image of the canon of novel theory that emerges from Michael McKeon's Theory of the Novel: A Historical Approach and Dorothy J. Hale's, The Novel: An Anthology of Criticism and Theory, 1900–2000 before moving to a longer consideration of Franco Moretti's massive two‐volume collection The Novel. The essay explores how the conclusions of the ambitious founding texts of Western novel theory (by writers like Mikhail Bakhtin and Georg Lukács, in particular) are revisited and revised in Moretti's collection, with its vast historical reach and its collection of materials from a large number of languages. These anthologies, despite their differences, suggest that the novel (as it is portrayed in novel theory at least) remains the paradigmatic form of modernity – and that novel criticism remains a crucial critical name for the ambition to come to grips with the contours of an increasingly globalized cultural scene.
A common conformationally coupled ATPase mechanism for yeast and human cytoplasmic HSP90s
Authors: Vaughan Cara K. , Piper Peter W. , Pearl Laurence H. , Prodromou Chrisostomos
Published: November 2008

The conformationally coupled mechanism by which ATP is utilized by yeast Hsp90 is now well characterized. In contrast, ATP utilization by human Hsp90s is less well studied, and appears to operate differently. To resolve these conflicting models, we have conducted a side‐by‐side biochemical analysis in a series of mutant yeast and human Hsp90s that have been both mechanistically and structurally characterized with regard to the crystal structure of the yeast Hsp90 protein. We show that each monomer of the human Hsp90 dimer is mutually dependent on the other for ATPase activity. Fluorescence studies confirmed that the N‐terminal domains of Hsp90β come into close association with each other. Mutations that directly affect the conformational dynamics of the ATP‐lid segment had marked effects, with T31I (yeast T22I) and A116N (yeast A107N) stimulating, and T110I (yeast T101I) inhibiting, human and yeast ATPase activity to similar extents, showing that ATP‐dependent lid closure is a key rate‐determining step in both systems. Mutation of residues implicated in N‐terminal dimerization of yeast Hsp90 (L15R and L18R in yeast, L24R and L27R in humans) significantly reduced the ATPase activity of yeast and human Hsp90s, showing that ATP‐dependent association of the N‐terminal domains in the Hsp90 dimer is also essential in both systems. Furthermore, cross‐linking studies of the hyper‐active yeast A107N and human A116N ATP‐lid mutants showed enhanced dimerization, suggesting that N‐terminal association is a direct consequence of ATP binding and lid closure in both systems.
Maternal characteristics and feelings of distress resulting from the crying of high‐risk infants
Authors: TABUCHI Noriko , SHIMADA Keiko
Published: November 2008

Aim:  To analyze the extent of and factors associated with the range of mothers' feelings when they experienced the crying of their high‐risk (HR) infant.

Methods:  A cross‐sectional design was used. A mail survey yielded 127 mothers of HR infants from 61 institutions. The instruments were the: (i) Maternal Distress Score to measure feelings of maternal distress during the infant's crying scale; (ii) Maternal Emotion Scale; (iii) Japanese Beck Depression Inventory – Second Edition; (iv) Visual Analog Scale for the tendency toward anxiety; and (v) a demographic form. The data analysis used descriptive and inferential statistics.

Results:  There were no significant differences in maternal distress between the mothers of HR infants and those with normal infants. The high‐scoring items for feelings of difficulty in descending order were: “feeling of a gap from life before pregnancy”, “anxious about child‐care prospects for the future”, and “feeling of stress when the infant cries”. The low‐scoring items were: “child care is not enjoyable” and “I want to feel free when I hear the infant crying”. The factors relating to the mother's distress over her crying HR infant showed a significant positive correlation with the mother's tendency toward anxiety and with the effect of the infant's crying on others, infant feeding, other family members in the same residence, free time, emotions when hearing the infant crying, and feelings of depression.

Conclusion:  The emergence of these three high‐scoring items show promise for the further refinement of this multidimensional approach to understanding the range of responses experienced by mothers with HR infants. The results are useful for understanding the psychological tendencies of mothers with HR infants and, possibly, in the future might aid in the screening of mothers for those attributes.