bluuebunny/crossref_metadata_2025 huggingface dataset:
Responses to cafeteria feeding in mice after the removal of interscapular brown adipose tissue
Authors: Connolly E. , Carnie J. A.
Published: February 2005

Feeding acafeteria diet to mice resulted in an increased energy intake of approximately 30% and this led to increases in the wet weight, total protein content, and total cytochrome oxidase activity of interscapular and dorso-cervical brown adipose tissue. Surgical removal of interscapular brown adipose tissue, followed by cafeteria feeding, gave rise to an elevation in dorso-cervical brown adipose tissue wet weight, total protein content, and total cytochrome oxidase activity, compared to intact cafeteria-fed mice. Cafeteria feeding with or without the removal of interscapular brown adipose tissue did not lead to significant increases in body weight compared to stock-fed control mice, but both cafeteria-fed groups of mice showed significant elevations in body fat content indicating that the induced hyperphagia led to a relative obesity in the cafeteria-fed groups. The results presented are consistent with an increased thermogenic activity in the brown adipose tissue of cafeteria-fed mice, and the effect of the removal of interscapular brown adipose tissue further indicates the quantitative importance of the tissue in the control of body weight.
Immobilization of enzymes on alumina by means of pyridoxal 5′-phosphate
Authors: Pugnière M. , San Juan C. , Coletti-Previero M-A. , Previero A.
Published: February 2005

A number of proteases have been immobilized on alumina in a two-step procedure: the first step converted them into semisynthetic phosphoproteins which, in the second step, spontaneously bonded to alumina through their phosphate function. The immobilized enzymes thus obtained showed the physical properties typical of the inorganic carrier and a high activity on low molecular weight substrates.
Alterations in antigenic structure of gonadotropins following deglycosylation
Authors: Sairam M. R. , Linggen J. , Bhargavi G. N.
Published: February 2005

Radioimmunological techniques were utilized to probe possible changes in conformation of gonadotropins (human chorionic gonadotropin-hCG; and ovine luteinizing hormone—oLH) following chemical deglycosylation (DG-hCG and DG-LH). All antisera produced in rabbits, rats or mice contained antibodies that were specific to the deglycosylated hormones with the native hormones showing weak and non-parallel cross-reaction (<5%), but with rabbit antibodies to native hormones the deglycosylated hormones were fully reactive. Using hCG, asialo-hCG (A-hCG) and DG-hCG, we have shown that removal of sugars internal to sialic acid is required to produce these specific antibodies. These are in complete agreement with the observations that extensive deglycosylation of these hormones is necessary to induce changes in biological activity at the cellular level. Based on these data, we suggest that chemical deglycosylation results in changes in antigenic structure of these hormones by generation of new determinants or exposure of previously buried sites and these changes are of no consequence to receptor recognition.
Immunological identification of two lamina-like proteins in Saccharomyces cerevisiae
Authors: Galcheva-Gargova Zoya , Stateva Lubomira
Published: February 2005

Proteins from Saccharomyces cerevisiae were tested for their crossreactivity with antibodies raised against nuclear lamina proteins of Ehrlich Ascites Tumour cells. The results of the immunoblotting experiments and ELISA suggest the existence of at least two proteins (65 and 59 kDa) which are immunologically related to the nuclear lamina proteins of higher eukaryotes.
Purine nucleotide cycle enzymes in dystrophic and normal mouse muscle
Authors: Fitt Peter S. , Parliament Matthew B.
Published: February 2005

A comparative study of the operation of the purine nucleotide cycle and of the activity of adenylosuccinase in extracts of muscle from the two strains of dystrophic mouse shows that the cycle is defective in both cases in the conversion of adenylosuccinate to AMP. However, adenylosuccinase activity is only slightly reduced in the standard conditions for its direct assay.
In vitro transcription of viroid RNA into full-length copies by RNA-dependent RNA polymerase from healthy tomato leaf tissue
Authors: Boege Frank , Rohde Wolfgang , Sänger Heinz L.
Published: February 2005

RNA-dependent RNA polymerase from healthy tomato plant tissue accepts potato spindle tuber viroid (PSTV) RNA as a template for the in vitro synthesis of full-length RNA copies of the PSTV genome. Viroid transcription requires the presence of Mn2+ and/or Mg2+ ions and is not inhibited by concentrations of 10−5 M α-amanitin. This is the first report of a well-defined product synthesized in vitro by an RNA-dependent RNA polymerase from healthy plants.
Chicken liver fatty acid synthetase dimer: Evidence of asymmetrical structure from iodoacetamide inhibition studies
Authors: Kumar Suriender , Varagiannis Evelyn
Published: February 2005

The condensing component of chicken liver fatty acid synthetase is inhibited by a sulfhydryl reagent, iodoacetamide, with a second-order rate constant of 0.23 M−1 sec−1 at pH 7.0 and 0°. Complete inactivation requires the modification of approximately 8-SH groups per dimer of the enzyme. Quantitation of the extent of inactivation in the presence of i mM acetyl CoA (which completely protects the enzyme against inactivation) and in its absence shows that complete inactivation results from the binding of approximately 1.1 tool of carboxamidomethyl residues per dimer. These data are consistent with the proposed functional asymmetry of the enzyme.
Nuclear accumulation of a heat-shock 70-like protein during herpes simplex virus replication
Authors: LaThangue Nicholas B. , Latchman David S.
Published: February 2005

A monoclonal antibody defines an antigen, p68, related to hsp70, which is located in nuclei of uninfected exponential cells. Nuclear p68 is released by DNase but not RNase treatment suggesting an association with DNA. Lytic productive infection of confluent quiescent BHK 21 cells with herpes simplex virus type-2 causes p68 to accumulate in nuclei. The effect is specific for HSV-2, and does not occur in HSV-1 infected cells. Maximum nuclear accumulation of p68 requires virus DNA synthesis although a significant accumulation occurs in the absence of such synthesis. It is suggested that the nuclear accumulation of p68 is an aspect of a cellular stress response to lytic infection with HSV-2.
A new protein (J1-31 antigen, 30 kD) is expressed by astrocytes, Müller glia and ependyma
Authors: Predy R. , Singh D. , Bhatnagar R. , Singh R. , Malhotra S. K.
Published: February 2005

Monoclonal antibody (MAb) J1–31 raised using human brain homogenate as immunogen in mice can be used as a cell type marker for certain types of CNS macroglia, namely astrocytes, Müller cells and tanycytes as well as ciliated ependymal cells. Except for the ciliated ependymal cells, these types of macroglia express glial fibrillary acidic protein (GFAP). J1–31 antigen is an intracellular protein which has a MW of 30 kD under reducing conditions for gel electrophoresis (Singh et al., 1986). This protein is distinct from GFAP (MW 50 kD) and vimentin (MW 55 kD), the two core proteins of 10 nm IFs known to be expressed in the above types ofmacroglia. This conclusion is based on several criteria including temporal differences in the onset of expression of GFAP and J1–31 antigen during development of the rat cerebellum. Also, there is no detectable (by immunofluorescence microscopy) expression of J1–31 antigen in the prenatal CNS or outside the CNS where vimentin has been reported to be abundant. The most direct evidence that J 1–31 antigen and GFAP are distinct proteins comes from studies on the mature ciliated ependymal cells which do not express GFAP and yet show intense immunostaining for J1–31 antigen.
Clenbuterol, a beta agonist, induces growth in innervated and denervated rat soleus muscle via apparently different mechanisms
Authors: Maltin C. A. , Hay S. M. , Delday M. I. , Smith F. G. , Lobley G. E. , Reeds P. J.
Published: February 2005

Dietary administration of the anabolic agent, clenbuterol, has already been shown to inhibit or reverse denervation-induced atrophy in rat soleus muscles. We now show that the ameliorative effects of clenbuterol in denervated rat muscles are due principally to a large increase in protein synthesis. This results from both an increase in protein synthetic capacity and a normalised translational efficiency. The responses of innervated and denervated muscles are therefore fundamentally different, the changes in denervated muscles being reminiscent of the classical pleiotypic response of cells to growth factors.
Gene Overexpression in Pancreatic Adenocarcinoma: Diagnostic and Therapeutic Implications
Authors: Rodriguez Joel A. , Li Min , Yao Qizhi , Chen Changyi , Fisher William E.
Published: February 2005

Pancreatic cancer is the deadliest of cancers, and effective diagnostic and therapeutic strategies are lacking. Global gene expression profiling holds promise for improved diagnosis and treatment. Knowledge of the location and timing of gene overexpression and the function of these genes, including their effects on signaling pathways, is important. These data may be used to develop histologic and serum biomarkers as well as to develop immunotherapeutic, molecular targeting, and gene therapy strategies. We have compiled a list of overexpressed genes in pancreatic cancer for which overexpression was confirmed by reverse transcriptase polymerase chain reaction, immunohistochemistry, and/or in situ hybridization following initial identification by global gene expression profiling. The techniques used in the determination of overexpression, problems encountered in global gene expression profiling, and the diagnostic and therapeutic implications of overexpression are discussed. The S100 gene family, mesothelin, prostate stem cell antigen, and 14‐3‐3 sigma, may have important clinical implications in pancreatic cancer diagnosis and treatment.
Bystander effect of normal fibroblasts for macrophages co‐cultured with susceptible transformed target cells
Authors: Kashchak Nataliya , Tsaryk Roman , Stoika Rostyslav
Published: February 2005

Macrophages attack and kill pathologically changed, transformed and tumor cells. However, in some cases they may also support tumor growth, modulate the action of anticancer drugs, and even facilitate the development of drug resistance in tumor cells. Here we present data that bystander fibroblasts NIH3T3 were not only resistant to murine macrophages J774.2 but also blocked their killing action towards murine transformed fibroblasts L929. Macrophages were isolated from mixed cultures by means of CD11b specific immunomagnetic beads, and changes induced by their former co‐culturing were studied using DNA microarray technology and other tests. An expression of candidate genes coding for cytokines and for signal transduction pathway proteins was estimated in macrophages in different variants of their co‐culture with target cells. Changes in expression of mRNA for interleukin 1β, NFκB, IκBα, gadd45, and CD5 were detected as the most prominent in the macrophages co‐cultured with the transformed cells. Bystander NIH3T3 fibroblasts abolished these changes in the macrophages J774.2, and the level of expression of the above mentioned genes was close to the level seen in the macrophages which did not exert cytotoxicity towards the target fibroblasts. Potential implications and research perspectives of using the macrophage—target cell co‐cultures with different bystander cellular partners are discussed.
For Better and For Worse: Quality of Life Impaired in HF Patients as well as in Their Partners
Authors: Luttik Marie Louise , Jaarsma Tiny , Veeger Nie J.G.M. , van Veldhuisen Dirk J.
Published: February 2005

Background: Quality of Life (QOL) is known to be impaired in patients with Heart Failure (HF). The involvement of a key person, most often the spouse, enables the HF patient to manage the medical regimen and therefore to sustain Quality of Life (QOL). Yet little is known on the impact of caring for an HF patient on the QOL of the caregiving partner. This study aims to explore the QOL of partners of HF patients compared to the QOL of the patients.

Methods: The study population consisted of 38 couples of hospitalized HF patients and their partners. The Cantril Ladder of Life was used to rate QOL during hospitalization, with regard to the month prior to hospitalization and as projected 3 years in the future. Demographic and clinical variables were collected by patient interview and chart review.

Results: On a scale from 0–10, QOL scores of partners varied from 5.9 to 6.4. At some point the QOL score of partners was even lower than the QOL scores of HF patients. In the month prior to hospital admission the QOL of partners was significantly higher in comparison to the QOL of HF patients (6.1 vs. 4.9, respectively). However, this reversed during hospital admission, with QOL scores of partners being significantly lower compared to QOL scores of HF patients (5.9 vs. 6.8, respectively), even after correcting for age and gender.

Conclusion: In our study the QOL of partners of HF patients was low. Whether this is explicitly due to having to live with a HF patient is not clear. Further research on what partners actually do and the relationship between being a caregiver and QOL is necessary in order to support these partners in giving optimal care and support.
Self-rated general health and psychiatric disorders in a general population sample
Authors: John Ulrich , Meyer Christian , Rumpf Hans-Jürgen , Schumann Anja , Dilling Horst , Hapke Ulfert
Published: February 2005

The purpose of this study was to explore the relationships between nicotine and alcohol dependence, depressive, anxiety and somatoform disorders with self-rated general health (GH). A cohort study of a random sample of the non-institutionalised general population aged 18–64 with a participation rate of 70.2% was carried out in a German area (n = 4075 at baseline). A follow-up of tobacco smokers or heavy drinkers (n = 1083, 79.4% of those who had given consent to be followed-up) was conducted 30 months after baseline measurement. The assessments included self-ratings of GH and Diagnostic and Statistical Manual (DSM-IV) diagnoses based on the Composite International Diagnostic Interview. The results show that nicotine dependence, anxiety disorders and somatoform disorders moderately predicted self-rated GH at follow-up (general linear model, R2 = 0.12). We conclude that psychiatric disorders may contribute to the prediction of a low self-rated GH.
Depression and anxiety complaints; relations with sleep disturbances
Authors: Spoormaker Victor I. , van den Bout Jan
Published: February 2005

Objectives were to assess the relations of various sleep complaints with depressive and anxiety complaints in a non-clinical population. Four-hundred-and-two randomly approached adults received three questionnaires. Results showed a high interrelatedness between sleep and depressive/anxiety complaints. Both assessment and treatment of depressive and anxiety complaints should address sleep problems.
Moroccan colloquial Arabic version of the Mini International Neuropsychiatric Interview (MINI): qualitative and quantitative validation
Authors: Kadri N. , Agoub M. , Gnaoui S. El , Alami Kh. Mchichi , Hergueta T. , Moussaoui D.
Published: February 2005

The validation of mini international neuropsychiatric interview (MINI) into Moroccan Colloquial Arabic language demonstrated good psychometric properties. The concordance between translated MINI’s and expert diagnoses was good with kappa values greater than 0.80. The reliability inter-rater and test–retest were excellent with kappa values above 0.80 and 0.90, respectively.
A plasma membrane H
+
-ATPase is required for the formation of proanthocyanidins in the seed coat endothelium of Arabidopsis thaliana
Authors: Baxter Ivan R. , Young Jeffery C. , Armstrong Gordon , Foster Nathan , Bogenschutz Naomi , Cordova Tatiana , Peer Wendy Ann , Hazen Samuel P. , Murphy Angus S. , Harper Jeffrey F.
Published: February 2005

The plasma membrane in plant cells is energized with an electrical potential and proton gradient generated through the action of H
+
pumps belonging to the P-type ATPase superfamily. The
Arabidopsis
genome encodes 11 plasma membrane H
+
pumps. Auto-inhibited H
+
-ATPase isoform 10 (AHA10) is expressed primarily in developing seeds. Here we show that four independent gene disruptions of
AHA10
result in seed coats with a
transparent testa
(
tt
) phenotype (light-colored seeds). A quantitative analysis of extractable flavonoids in
aha10
seeds revealed an ≈100-fold reduction of proanthocyanidin (PA), one of the two major end-product pigments in the flavonoid biosynthetic pathway. In wild-type seed coat endothelial cells, PA accumulates in a large central vacuole. In
aha10
mutants, the formation of this vacuole is impaired, as indicated by the predominance of multiple small vacuoles observed by fluorescence microscopy using a vacuole-specific dye, 5-(and -6)-carboxy 2′,7′-dichlorofluorescein diacetate. A similar vacuolar defect was also observed for another
tt
mutant,
tt12
, a proton-coupled multidrug and toxic compound extrusion transporter potentially involved in loading provacuoles with a flavonoid intermediate required for PA production. The endothelial cells in
aha10
mutants are otherwise healthy, as indicated by the lack of a significant decrease in (
i
) the accumulation of other flavonoid pathway end products, such as anthocyanins, and (
ii
) mRNA levels for two endothelium-specific transcripts (
TT12
and
BAN
). Thus, the specific effect of
aha10
on vacuolar and PA biogenesis provides genetic evidence to support an unexpected endomembrane function for a member of the plasma membrane H
+
-ATPase family.
Force mapping in epithelial cell migration
Authors: du Roure Olivia , Saez Alexandre , Buguin Axel , Austin Robert H. , Chavrier Philippe , Siberzan Pascal , Ladoux Benoit
Published: February 2005

We measure dynamic traction forces exerted by epithelial cells on a substrate. The force sensor is a high-density array of elastomeric microfabricated pillars that support the cells. Traction forces induced by cell migration are deduced from the measurement of the bending of these pillars and are correlated with actin localization by fluorescence microscopy. We use a multiple-particle tracking method to estimate the mechanical activity of cells in real time with a high-spatial resolution (down to 2 μm) imposed by the periodicity of the post array. For these experiments, we use differentiated Madin-Darby canine kidney (MDCK) epithelial cells. Our data provide definite information on mechanical forces exerted by a cellular assembly. The maximum intensity of the forces is localized on the edge of the epithelia. Hepatocyte growth factor promotes cell motility and induces strong scattering activity of MDCK cells. Thus, we compare forces generated by MDCK cells in subconfluent epithelia versus isolated cells after hepatocyte growth factor treatment. Maximal-traction stresses at the edge of a monolayer correspond to higher values than those measured for a single cell and may be due to a collective behavior.
Stretched exponential decay and correlations in the catalytic activity of fluctuating single lipase molecules
Authors: Flomenbom Ophir , Velonia Kelly , Loos Davey , Masuo Sadahiro , Cotlet Mircea , Engelborghs Yves , Hofkens Johan , Rowan Alan E. , Nolte Roeland J. M. , Van der Auweraer Mark , de Schryver Frans C. , Klafter Joseph
Published: February 2005

Single-molecule techniques offer a unique tool for studying the dynamical behavior of individual molecules and provide the possibility to construct distributions from individual events rather than from a signal stemming from an ensemble of molecules. In biological systems, known for their complexity, these techniques make it possible to gain insights into the detailed spectrum of molecular conformational changes and activities. Here, we report on the direct observation of a single lipase-catalyzed reaction for extended periods of time (hours), by using confocal fluorescence microscopy. When adding a profluorescent substrate, the monitored enzymatic activity appears as a trajectory of “on-state” and “off-state” events. The waiting time probability density function of the off state and the state-correlation function fit stretched exponentials, independent of the substrate concentration in a certain range. The data analysis unravels oscillations in the logarithmic derivative of the off-state waiting time probability density function and correlations between off-state events. These findings imply that the fluctuating enzyme model, which involves a spectrum of enzymatic conformations that interconvert on the time scale of the catalytic activity, best describes the observed enzymatic activity. Based on this model, values for the coupling and reaction rates are extracted.
Quantitative trait locus mapping and DNA array hybridization identify an
FLM
deletion as a cause for natural flowering-time variation
Authors: Werner Jonathan D. , Borevitz Justin O. , Warthmann Norman , Trainer Gabriel T. , Ecker Joseph R. , Chory Joanne , Weigel Detlef
Published: February 2005

Much of the flowering time variation in wild strains of
Arabidopsis thaliana
is due to allelic variation at two epistatically acting loci,
FRIGIDA
(
FRI
) and
FLOWERING LOCUS C
(
FLC
).
FLC
encodes a MADS (MCM1/AGAMOUS/DEFICIENS/SRF1) domain transcription factor that directly represses a series of flowering-promoting genes.
FRI
and
FLC
, however, do not explain all of the observed variation, especially when plants are grown in short days. To identify loci that act in addition to
FRI
and
FLC
in controlling flowering of natural accessions, we have analyzed a recombinant inbred line population derived from crosses of accession Niederzenz (Nd) to Columbia, both of which contain natural
FRI
lesions. Quantitative trait locus mapping and genomic DNA analysis by microarray hybridization were used to identify candidate genes affecting variation in flowering behavior. In both long and short days, the quantitative trait locus of largest effect, termed
FLOWERING 1
(
FLW1
), was found to be associated with a Nd-specific deletion of
FLOWERING LOCUS M
(
FLM
), which encodes a floral repressor closely related to
FLC
. Analysis of near isogenic lines and quantitative transgenic complementation experiments confirmed that the
FLM
deletion is, in large part, responsible for the early flowering of the Nd accession.
Pituitary adenylate cyclase-activating polypeptide prevents the effects of ceramides on migration, neurite outgrowth, and cytoskeleton remodeling
Authors: Falluel-Morel Anthony , Vaudry David , Aubert Nicolas , Galas Ludovic , Benard Magalie , Basille Magali , Fontaine Marc , Fournier Alain , Vaudry Hubert , Gonzalez Bruno J.
Published: February 2005

During neuronal migration, cells that do not reach their normal destination or fail to establish proper connections are eliminated through an apoptotic process. Recent studies have shown that the proinflammatory cytokine tumor necrosis factor α (and its second messengers ceramides) and the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) play a pivotal role in the histogenesis of the cerebellar cortex. However, the effects of ceramides and PACAP on migration of cerebellar granule cells have never been investigated. Time-lapse videomicroscopy recording showed that C2-ceramide, a cell-permeable ceramide analog, and PACAP induced opposite effects on cell motility and neurite outgrowth. C2-ceramide markedly stimulated cell movements during the first hours of treatment and inhibited neuritogenesis, whereas PACAP reduced cell migration and promoted neurite outgrowth. These actions of C2-ceramide on cell motility and neurite outgrowth were accompanied by a disorganization of the actin filament network, depolarization of tubulin, and alteration of the microtubule-associated protein Tau. In contrast, PACAP strengthened the polarization of actin at the emergence cone, increased Tau phosphorylation, and abolished C2-ceramide-evoked alterations of the cytoskeletal architecture. The caspase-inhibitor Z-VAD-FMK, like PACAP, suppressed the “dance of the death” provoked by C2-ceramide. Finally, Z-VAD-FMK and the PP2A inhibitor okadaic acid both prevented the impairment of Tau phosphorylation induced by C2-ceramide. Taken together, these data indicate that the reverse actions of C2-ceramide and PACAP on cerebellar granule cell motility and neurite outgrowth are attributable to their opposite effects on actin distribution, tubulin polymerization, and Tau phosphorylation.
Iron-Mediated Control of the Basic Helix-Loop-Helix Protein FER, a Regulator of Iron Uptake in Tomato
Authors: Brumbarova Tzvetina , Bauer Petra
Published: February 2005

Root iron mobilization genes are induced by iron deficiency downstream of an unknown signaling mechanism. The FER gene, encoding a basic helix-loop-helix domain protein and putative transcription factor, is required for induction of iron mobilization genes in roots of tomato (Lycopersicon esculentum). To study upstream regulatory events of FER action, we examined the control of FER gene and FER protein expression in response to iron nutritional status. We analyzed expression of the FER gene and FER protein in wild-type plants, in mutant plants with defects in iron uptake regulation, and in 35S transgenic plants that overexpressed the FER gene. An affinity-purified antiserum directed against FER epitopes was produced that recognized FER protein in plant protein extracts. We found that the FER gene and FER protein were consistently down-regulated in roots after generous (100 μ  m, physiologically optimal) iron supply compared to low (0.1 μ  m) and sufficient (10 μ  m) iron supply. FER gene and FER protein expression were also occasionally down-regulated at sufficient compared to low iron supply. Analysis of FER protein expression in FER overexpression plants, as well as cellular protein localization studies, indicated that FER was down-regulated by high iron at the posttranscriptional level. The FER protein was targeted to plant nuclei and showed transcriptional activation in yeast (Saccharomyces cerevisiae). FER protein regulation in the iron accumulation mutant chloronerva indicated that FER protein expression was not directly controlled by signals derived from iron transport. We conclude that FER is able to affect transcription in the nucleus and its action is controlled by iron supply at multiple regulatory levels.
Cardiac-specific blockade of NF-κB in cardiac pathophysiology: differences between acute and chronic stimuli in vivo
Authors: Brown Maria , McGuinness Michael , Wright Terry , Ren Xiaoping , Wang Yang , Boivin Gregory P. , Hahn Harvey , Feldman Arthur M. , Jones W. Keith
Published: February 2005

The role of NF-κB in cardiac physiology and pathophysiology has been difficult to delineate due to the inability to specifically block NF-κB signaling in the heart. Cardiac-specific transgenic models have recently been developed that repress NF-κB activation by preventing phosphorylation at specific serine residues of the inhibitory κB (IκB) protein isoform IκBα. However, these models are unable to completely block NF-κB because of a second signaling pathway that regulates NF-κB function via Tyr42 phosphorylation of IκBα. We report the development of transgenic (3M) mouse lines that express the mutant IκBα(S32A,S36A,Y42F)in a cardiac-specific manner. NF-κB activation in cardiomyopathic TNF-1.6 mice is completely blocked by the 3M transgene but only partially blocked (70–80%) by the previously described double-mutant 2M [IκBα(S32A,S36A)] transgene, which demonstrates the action of two proximal pathways for NF-κB activation in TNF-α-induced cardiomyopathy. In contrast, after acute stimuli including administration of TNF-α and ischemia-reperfusion (I/R), NF-κB activation is blocked in both 2M and 3M transgenic mice. This result suggests that phosphorylation of the regulatory Ser32 and Ser36 predominantly mediates NF-κB activation in these situations. We show that infarct size after I/R is reduced by 70% in 3M transgenic mice, which conclusively demonstrates that NF-κB is involved in I/R injury. In summary, we have engineered novel transgenic mice that allow us to distinguish two major proximal pathways for NF-κB activation. Our results demonstrate that the serine and tyrosine phosphorylation pathways are differentially activated during different pathophysiological processes (cardiomyopathy and I/R injury) and that NF-κB contributes to infarct development after I/R.
Erythrocyte-associated transients in Po2revealed in capillaries of rat mesentery
Authors: Golub Aleksander S. , Pittman Roland N.
Published: February 2005

Mathematical models have predicted the existence of Po2gradients between erythrocytes in capillaries in the usual case where plasma contributes substantial resistance to oxygen diffusion. According to theoretical predictions, these gradients could be detected as rapid Po2fluctuations (erythrocyte-associated transients, EATs) along the capillary. However, verification of a model and correct choice of its parameters can be made only on the basis of direct experimental measurements. We used phosphorescence quenching microscopy to measure Po2in 52 capillaries of rat mesentery to obtain plasma Po2values 100 times/s at a given point along a capillary. A 532-nm laser generated 10-μs pulses of light, concentrated by a ×100 objective, onto a spot 0.9 μm in diameter. The presence of erythrocytes in the excitation region was detected on the basis of phosphorescence amplitude (PA), proportional to the amount of plasma encountered by the laser beam, and on the basis of the intensity of transmitted laser light (LT), detected by a photodiode placed under the capillary. The data revealed correlated waveforms in PA, LT, and Po2in capillaries. The magnitude of the Po2gradients between erythrocytes and plasma was correlated with average capillary Po2. EATs in Po2were more readily detected in capillaries with relatively low oxygenation. The correlation coefficients between PA and Po2for the half of the capillaries ( n = 26) below the median Po2(mean Po2= 17 mmHg; R = −0.72) was higher than that for the other half (mean Po2= 39 mmHg; R = −0.38). These results support the theoretical predictions of EATs and plasma Po2gradients in capillaries.
U-46619 but not serotonin increases endocannabinoid content in middle cerebral artery: evidence for functional relevance
Authors: Rademacher David J. , Patel Sachin , Ho W.-S. Vanessa , Savoie Amanda M. , Rusch Nancy J. , Gauthier Kathryn M. , Hillard Cecilia J.
Published: February 2005

Cerebral vascular smooth muscle cells express the CB1cannabinoid receptor, and CB1receptor agonists produce vasodilation of cerebral arteries. The purpose of this study was to determine whether vasoconstriction of rat middle cerebral artery (MCA) results in the local formation of endocannabinoids (eCBs), which, via activation of CB1receptors, oppose the vasoconstriction in a feedback manner. The thromboxane A2(TXA2) mimetic U-46619 significantly increased N-arachidonylethanolamine (AEA) and 2-arachidonylglycerol (2-AG) content of isolated MCA, whereas 5-hydroxytrypamine (5-HT) decreased AEA and 2-AG content. If eCBs play a feedback role in the regulation of MCA tone, then CB1receptor antagonists should enhance the constriction of MCA produced by U-46619 but not 5-HT. U-46619 caused concentration-dependent constrictions of endothelium-denuded MCA. Two CB1receptor antagonists SR-141716 and AM-251 decreased the EC50value for U-46619 to constrict endothelium-denuded MCA without affecting the maximal effect. A low concentration of CB1receptor agonist Win-55212-2 (30 nM) produced vasodilation of MCAs constricted with low but not saturating concentrations of U-46619. SR-141716 had no effect on the 5-HT concentration-contraction relationship. These data suggest that TXA2receptor activation increases MCA eCB content, which, via activation of CB1receptors, reduces the constriction produced by moderate concentrations of the TXA2agonist. Although 5-HT-induced vasoconstriction is reduced by exogenous CB1receptor agonist, activation of 5-HT receptors does not increase eCB content. These results suggest that MCA production of eCBs is not regulated by constriction per se but likely via a signaling pathway that is specific for TXA2receptors and not 5-HT receptors.
Podosome-mediated matrix resorption and cell motility in vascular smooth muscle cells
Authors: Burgstaller Gerald , Gimona Mario
Published: February 2005

The migration of vascular smooth muscle cells (VSMCs) is a principal factor for the development and progression of vascular diseases. In addition, phenotypic alteration from the contractile (differentiated) to the synthetic (dedifferentiated) state and a proteolytic process in the form of extra cellular matrix degradation are necessary for SMC invasion. The actual mechanism leading to the focal degradation of basement membrane matrix components and, hence, SMC migration within the tissue itself is, however, unclear. In response to phorbol ester [phorbol-12,13-dibutyrate (PDBu)], VSMCs in culture form podosomes, dynamic organelles critical for cell adhesion and substrate degradation that are typically found in invasive cells and cells that cross tissue boundaries. Here, we show that PDBu-stimulated VSMCs resorb the extracellular matrix at the sites of podosomes. Podosome formation correlates with an increased polarization of VSMCs on fibronectin- or collagen-coated flexible substrates in addition to a concomitant induction of cell motility. VSMCs embedded in reconstituted basement membrane support adopt the typical spindle-shaped morphology of differentiated SMCs in vivo and, after PDBu treatment, form peripheral lamellipodia and podosomes around their matrix-contacting surface. Our findings demonstrate that podosome formation is the potential mechanism underlying the ability of VSMCs to traverse the surrounding basement membrane and escape the barrier of the tunica media in vascular diseases.
Thermomechanical behavior of human carotid arteries in the passive state
Authors: Guinea G. V. , Atienza J. M. , Elices M. , Aragoncillo P. , Hayashi K.
Published: February 2005

Localized heating or cooling is expanding the clinical procedures used to treat cardiovascular diseases. Advantageous implementation and development of these methods are linked indissolubly to a deeper understanding of the arterial response to combined mechanical and thermal loads. Despite this, the basic thermomechanical behavior of human blood vessels still remains largely unknown, primarily due to the lack of appropriate experimental data. In this work, the influence of temperature on the passive behavior of human carotid arteries was studied in vitro by means of inflation tests. Eleven carotid segments were tested in the range 0–200 mmHg at four different temperatures of 17, 27, 37, and 42°C. The results show that the combined change of temperature and stress has a dramatic effect on the dilatation coefficient of the arterial wall, which is shifted from negative to positive depending on the stress state, whereas the structural stiffness of the arterial wall does not change appreciably in the range of temperatures tested.
Oxygen increases ductus arteriosus smooth muscle cytosolic calcium via release of calcium from inositol triphosphate-sensitive stores
Authors: Keck Maggie , Resnik Ernesto , Linden Bradley , Anderson Franklin , Sukovich David J. , Herron Jean , Cornfield David N.
Published: February 2005

In utero, blood shunts away from the lungs via the ductus arteriosus (DA) and the foramen ovale. After birth, the DA closes concomitant with increased oxygen tension. The present experimental series tests the hypothesis that oxygen directly increases DA smooth muscle cell (SMC) cytosolic calcium ([Ca2+]i) through inactivation of a K+channel, membrane depolarization, and entry of extracellular calcium. To test the hypothesis, DA SMC were isolated from late-gestation fetal lambs and grown to subconfluence in primary culture in low oxygen tension (25 Torr). DA SMC were loaded with the calcium-sensitive fluorophore fura-2 under low oxygen tension conditions and studied using microfluorimetry while oxygen tension was acutely increased (120 Torr). An acute increase in oxygen tension progressively increased DA SMC [Ca2+]iby 11.7 ± 1.4% over 40 min. The effect of acute normoxia on DA SMC [Ca2+]iwas mimicked by pharmacological blockade of the voltage-sensitive K+channel. Neither removal of extracellular calcium nor voltage-operated calcium channel blockade prevented the initial increase in DA SMC [Ca2+]i. Manganese quenching experiments demonstrated that acute normoxia initially decreases the rate of extracellular calcium entry. Pharmacological blockade of inositol triphosphate-sensitive, but not ryanodine-sensitive, intracellular calcium stores prevented the oxygen-induced increase in [Ca2+]i. Endothelin increased [Ca2+]iin acutely normoxic, but not hypoxic, DA SMC. Thus acute normoxia 1) increases DA SMC [Ca2+]ivia release of calcium from intracellular calcium stores, and subsequent entry of extracellular calcium, and 2) potentiates the effect of contractile agonists. Prolonged patency of the DA may result from disordered intracellular calcium homeostasis.
Memorization and Association on a Realistic Neural Model
Authors: Valiant Leslie G.
Published: February 2005

A central open question of computational neuroscience is to identify the data structures and algorithms that are used in mammalian cortex to support successive acts of the basic cognitive tasks of memorization and association. This letter addresses the simultaneous challenges of realizing these two distinct tasks with the same data structure, and doing so while respecting the following four basic quantitative parameters of cortex: the neuron number, the synapse number, the synapse strengths, and the switching times. Previous work has not succeeded in reconciling these opposing constraints, the low values of synapse strengths that are typically observed experimentally having contributed a particular obstacle. In this article, we describe a computational scheme that supports both memory formation and association and is feasible on networks of model neurons that respect the widely observed values of the four quantitative parameters. Our scheme allows for both disjoint and shared representations. The algorithms are simple, and in one version both memorization and association require just one step of vicinal or neighborly influence. The issues of interference among the different circuits that are established, of robustness to noise, and of the stability of the hierarchical memorization process are addressed. A calculus therefore is implied for analyzing the capabilities of particular neural systems and subsystems, in terms of their basic numerical parameters.
Maximum Likelihood Topographic Map Formation
Authors: Hulle Marc M. Van
Published: February 2005

We introduce a new unsupervised learning algorithm for kernel-based topographic map formation of heteroscedastic gaussian mixtures that allows for a unified account of distortion error (vector quantization), log-likelihood, and Kullback-Leibler divergence.
The Puzzle of System Transformation: Towards an Explanatory Sketch
Authors: Zimmermann Ekkart
Published: February 2005

After the collapse of communism in Eastern Europe, there is renewed interest in comparative studies of system transformation and revolution. System transformation must be delineated from revolution as an outcome. Revolutionary situations are one independent causal variable, but not a definiens of system transformation. Our definition of system transformation as the substitution of previous fundamental political and economic institutions in a polity by new (or different) political and economic institutions is relatively short and points to several typological cases of system transformation. Our causal model of system transformation amounts to almost twenty causal links that can be sizably reduced into five causal bloc variables: state breakdown, defeat in war and external conquest, economic growth, great power rivalry, and natural catastrophes.
Do crimes against humanity deserve a higher sentence than war crimes?
Authors: Olusanya Olaoluwa Abiola
Published: February 2005

This article examines the issue of penalties for crimes against humanity and war crimes as imposed by the judges of the International Criminal Tribunal for the former Yugoslavia (ICTY). At the moment, if for example A murders B, C, and D in the course of an armed conflict, it appears to makes no difference in terms of penalties if he is ultimately convicted under crimes against humanity or war crimes, what appears to matter is the fact that he committed murder. This approach has the effect of obscuring the criminal intention and motives for war crimes and crimes against humanity. The main reason why the ICTY has persisted with this approach, is the fact that at present there appears to be no credible index for adopting a policy of relative sentencing for crimes against humanity and war crimes. The aim of this Article therefore is to advocate a method for imposing higher penalties for crimes against humanity vis-à-vis war crimes. Furthermore, this method will rely on the argument that crimes against humanity offences e.g. murder, rape and torture when committed with a discriminatory motive should attract higher penalties than identical offences when committed as war crimes.
Influence of language and ancestry on genetic structure of contiguous populations: A microsatellite based study on populations of Orissa
Authors: Sahoo Sanghamitra , Kashyap VK
Published: February 2005

Background

We have examined genetic diversity at fifteen autosomal microsatellite loci in seven predominant populations of Orissa to decipher whether populations inhabiting the same geographic region can be differentiated on the basis of language or ancestry. The studied populations have diverse historical accounts of their origin, belong to two major ethnic groups and different linguistic families. Caucasoid caste populations are speakers of Indo-European language and comprise Brahmins, Khandayat, Karan and Gope, while the three Australoid tribal populations include two Austric speakers: Juang and Saora and a Dravidian speaking population, Paroja. These divergent groups provide a varied substratum for understanding variation of genetic patterns in a geographical area resulting from differential admixture between migrants groups and aboriginals, and the influence of this admixture on population stratification.

Results

The allele distribution pattern showed uniformity in the studied groups with approximately 81% genetic variability within populations. The coefficient of gene differentiation was found to be significantly higher in tribes (0.014) than caste groups (0.004). Genetic variance between the groups was 0.34% in both ethnic and linguistic clusters and statistically significant only in the ethnic apportionment. Although the populations were genetically close (FST = 0.010), the contemporary caste and tribal groups formed distinct clusters in both Principal-Component plot and Neighbor-Joining tree. In the phylogenetic tree, the Orissa Brahmins showed close affinity to populations of North India, while Khandayat and Gope clustered with the tribal groups, suggesting a possibility of their origin from indigenous people.

Conclusions

The extent of genetic differentiation in the contemporary caste and tribal groups of Orissa is highly significant and constitutes two distinct genetic clusters. Based on our observations, we suggest that since genetic distances and coefficient of gene differentiation were fairly small, the studied populations are indeed genetically similar and that the genetic structure of populations in a geographical region is primarily influenced by their ancestry and not by socio-cultural hierarchy or language. The scenario of genetic structure, however, might be different for other regions of the subcontinent where populations have more similar ethnic and linguistic backgrounds and there might be variations in the patterns of genomic and socio-cultural affinities in different geographical regions.
Construction of a radiation hybrid map of chicken chromosome 2 and alignment to the chicken draft sequence
Authors: Leroux Sophie , Dottax Mélanie , Bardes Suzanne , Vignoles Florence , Fève Katia , Pitel Frédérique , Morisson Mireille , Vignal Alain
Published: February 2005

Background

The ChickRH6 whole chicken genome radiation hybrid (RH) panel recently produced has already been used to build radiation hybrid maps for several chromosomes, generating comparative maps with the human and mouse genomes and suggesting improvements to the chicken draft sequence assembly. Here we present the construction of a RH map of chicken chromosome 2. Markers from the genetic map were used for alignment to the existing GGA2 (Gallus gallus chromosome 2) linkage group and EST were used to provide valuable comparative mapping information. Finally, all markers from the RH map were localised on the chicken draft sequence assembly to check for eventual discordances.

Results

Eighty eight microsatellite markers, 10 genes and 219 EST were selected from the genetic map or on the basis of available comparative mapping information. Out of these 317 markers, 270 gave reliable amplifications on the radiation hybrid panel and 198 were effectively assigned to GGA2. The final RH map is 2794 cR6000 long and is composed of 86 framework markers distributed in 5 groups. Conservation of synteny was found between GGA2 and eight human chromosomes, with segments of conserved gene order of varying lengths.

Conclusion

We obtained a radiation hybrid map of chicken chromosome 2. Comparison to the human genome indicated that most of the 8 groups of conserved synteny studied underwent internal rearrangements. The alignment of our RH map to the first draft of the chicken genome sequence assembly revealed a good agreement between both sets of data, indicative of a low error rate.
Development of a cDNA array for chicken gene expression analysis
Authors: Burnside Joan , Neiman Paul , Tang Jianshan , Basom Ryan , Talbot Richard , Aronszajn Mark , Burt David , Delrow Jeff
Published: February 2005

Background

The application of microarray technology to functional genomic analysis in the chicken has been limited by the lack of arrays containing large numbers of genes.

Results

We have produced cDNA arrays using chicken EST collections generated by BBSRC, University of Delaware and the Fred Hutchinson Cancer Research Center. From a total of 363,838 chicken ESTs representing 24 different adult or embryonic tissues, a set of 11,447 non-redundant ESTs were selected and added to an existing collection of clones (4,162) from immune tissues and a chicken bursal cell line (DT40). Quality control analysis indicates there are 13,007 useable features on the array, including 160 control spots. The array provides broad coverage of mRNAs expressed in many tissues; in addition, clones with expression unique to various tissues can be detected.

Conclusions

A chicken multi-tissue cDNA microarray with 13,007 features is now available to academic researchers from genomics@fhcrc.org. Sequence information for all features on the array is in GenBank, and clones can be readily obtained. Targeted users include researchers in comparative and developmental biology, immunology, vaccine and agricultural technology. These arrays will be an important resource for the entire research community using the chicken as a model.