bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of alveolar epithelial function by hypoxia
Authors: Zhou G. , Dada L. A. , Sznajder J. I.
Published: April 2008

Patients with acute respiratory distress syndrome and high-altitude pulmonary oedema build up excess lung fluid, which leads to alveolar hypoxia. In patients with acute respiratory distress syndrome and hypoxia, there is a decrease in oedema fluid clearance, due in part to the downregulation of plasma membrane sodium–potassium adenosine triphosphatase (Na,K-ATPase).

In alveolar epithelial cells, acute hypoxia promotes Na,K-ATPase endocytosis from the plasma membrane to intracellular compartments, resulting in inhibition of Na,K-ATPase activity. Exposure to prolonged hypoxia leads to degradation of plasma membrane Na,K-ATPase.

The downregulation of plasma membrane Na,K-ATPase reduces adenosine triphosphate demand, as part of a survival mechanism of cellular adaptation to hypoxia. Hypoxia has also been shown to disassemble and degrade the keratin intermediate filament network, a fundamental component of the cell cytoskeleton, affecting epithelial barrier function.

Accordingly, better understanding of the mechanisms regulating cellular adaptation to hypoxia may lead to the development of novel therapeutic strategies for acute respiratory distress syndrome and high-altitude pulmonary oedema patients.
Epigallocatechin-3-gallate (EGCG) inhibits the migratory behavior of tumor bronchial epithelial cells
Authors: Hazgui Salma , Bonnomet Arnaud , Nawrocki-Raby Béatrice , Milliot Magali , Terryn Christine , Cutrona Jérôme , Polette Myriam , Birembaut Philippe , Zahm Jean-Marie
Published: April 2008

Background

Many studies associated the main polyphenolic constituent of green tea, (-)-Epigallocatechin-3-gallate (EGCG), with inhibition of cancers, invasion and metastasis. To date, most of the studies have focused on the effect of EGCG on cell proliferation or death. Since cell migration is an important mechanism involved in tumor invasion, the aim of the present work was to target another approach of the therapeutic effect of EGCG, by investigating its effect on the cell migratory behavior.

Methods

The effect of EGCG (at concentrations lower than 10 μg/ml) on the migration speed of invasive cells was assessed by using 2D and 3D models of cell culture. We also studied the effects of EGCG on proteinases expression by RT-PCR analysis. By immunocytochemistry, we analyzed alterations of vimentin organization in presence of different concentrations of EGCG.

Results

We observed that EGCG had an inhibitory effect of cell migration in 2D and 3D cell culture models. EGCG also inhibited MMP-2 mRNA and protein expression and altered the intermediate filaments of vimentin.

Conclusion

Taken together, our results demonstrate that EGCG is able to inhibit the migration of bronchial tumor cells and could therefore be an attractive candidate to treat tumor invasion and cell migration.
In vivo consequences of deleting EGF repeats 8–12 including the ligand binding domain of mouse Notch1
Authors: Ge Changhui , Liu Tongyi , Hou Xinghua , Stanley Pamela
Published: April 2008

Background

Notch signaling is highly conserved in the metazoa and is critical for many cell fate decisions. Notch activation occurs following ligand binding to Notch extracellular domain. In vitro binding assays have identified epidermal growth factor (EGF) repeats 11 and 12 as the ligand binding domain of Drosophila Notch. Here we show that an internal deletion in mouse Notch1 of EGF repeats 8–12, including the putative ligand binding domain (lbd), is an inactivating mutation in vivo. We also show that maternal and zygotic Notch1

lbd/lbd
mutant embryos develop through gastrulation to mid-gestation.

Results

Notch1

lbd/lbd
embryos died at mid-gestation with a phenotype indistinguishable from Notch1 null mutants. In embryonic stem (ES) cells, Notch1lbd was expressed on the cell surface at levels equivalent to wild type Notch1, but Delta1 binding was reduced to the same level as in Notch1 null cells. In an ES cell co-culture assay, Notch signaling induced by Jagged1 or Delta1 was reduced to a similar level in Notch1

lbd
and Notch1 null cells. However, the Notch1

lbd/lbd
allele was expressed similarly to wild type Notch1 in Notch1

lbd/lbd
ES cells and embryos at E8.75, indicating that Notch1 signaling is not essential for the Notch1 gene to be expressed. In addition, maternal and zygotic Notch1 mutant blastocysts developed through gastrulation.

Conclusion

Mouse Notch1 lacking the ligand binding domain is expressed at the cell surface but does not signal in response to the canonical Notch ligands Delta1 and Jagged1. Homozygous Notch1

lbd/lbd
mutant embryos die at ~E10 similar to Notch1 null embryos. While Notch1 is expressed in oocytes and blastocysts, Notch1 signaling via canonical ligands is dispensable during oogenesis, blastogenesis, implantation and gastrulation.
Gene Structure Evolution of the Na+-Ca2+ Exchanger (NCX) Family
Authors: On Caly , Marshall Christian R , Chen Nansheng , Moyes Christopher D , Tibbits Glen F
Published: April 2008

Background

The Na+-Ca2+ exchanger (NCX) is an important regulator of cytosolic Ca2+ levels. Many of its structural features are highly conserved across a wide range of species. Invertebrates have a single NCX gene, whereas vertebrate species have multiple NCX genes as a result of at least two duplication events. To examine the molecular evolution of NCX genes and understand the role of duplicated genes in the evolution of the vertebrate NCX gene family, we carried out phylogenetic analyses of NCX genes and compared NCX gene structures from sequenced genomes and individual clones.

Results

A single NCX in invertebrates and the protochordate Ciona, and the presence of at least four NCX genes in the genomes of teleosts, an amphibian, and a reptile suggest that a four member gene family arose in a basal vertebrate. Extensive examination of mammalian and avian genomes and synteny analysis argue that NCX4 may be lost in these lineages. Duplicates for NCX1, NCX2, and NCX4 were found in all sequenced teleost genomes. The presence of seven genes encoding NCX homologs may provide teleosts with the functional specialization analogous to the alternate splicing strategy seen with the three NCX mammalian homologs.

Conclusion

We have demonstrated that NCX4 is present in teleost, amphibian and reptilian species but has been secondarily and independently lost in mammals and birds. Comparative studies on conserved vertebrate homologs have provided a possible evolutionary route taken by gene duplicates subfunctionalization by minimizing homolog number.
Identification of novel vascular markers through gene expression profiling of tumor-derived endothelium
Authors: Ghilardi Carmen , Chiorino Giovanna , Dossi Romina , Nagy Zsuzsanna , Giavazzi Raffaella , Bani MariaRosa
Published: April 2008

Background

Targeting tumor angiogenesis and vasculature is a promising strategy for the inhibition of tumor growth and dissemination. Evidence suggests that tumor vasculature expresses unique markers that distinguish it from normal vasculature. Our efforts focused on the molecular characterization of endothelial cells (EC) in the search for selective markers of tumor vasculature that might be helpful for the development of effective therapeutic approaches.

Results

We investigated by microarray analysis the gene expression profiles of EC purified and cultured from tumor (ovarian carcinoma [HOC-EC]) and normal (human adrenal gland [HA-EC]) tissue specimens. We found distinct transcriptional features characterizing the EC of different origin, and identified 158 transcripts highly expressed by HOC-EC. We analyzed four of these genes, ADAM23, FAP, GPNMB and PRSS3, which were not previously known to be expressed by endothelium.In vitroexperiments confirmed the higher expression of the selected genes in tumor-derived endothelium with no expression in tumor cells.In vivoinvestigation byin situhybridization established that ADAM23, GPNMB and PRSS3 expression is localized on blood vessels of human cancer specimens.

Conclusion

These findings elucidate some of the molecular features of the tumor endothelium. Comparative transcriptomic analysis allowed us to determine molecular differences of tumor and normal tissue-derived endothelium and to identify novel markers that might be exploited to selectively target tumor vasculature.
Varicella-zoster virus ORF 58 gene is dispensable for viral replication in cell culture
Authors: Yoshii Hironori , Sadaoka Kay , Matsuura Masaaki , Nagaike Kazuhiro , Takahashi Michiaki , Yamanishi Koichi , Mori Yasuko
Published: April 2008

Background

Open reading frame 58 (ORF58) of varicella-zoster virus (VZV) lies at the 3'end of the Unique long (UL) region and its functional is unknown. In order to clarify whether ORF58 is essential for the growth of VZV, we constructed a deletion mutant of ORF58 (pOka-BACΔ58) from the Oka parental genome cloned into a bacterial artificial chromosome (pOka-BAC).

Results

The ORF58-deleted virus (rpOkaΔ58) was reconstituted from the pOka-BACΔ58 genome in MRC-5 cells, indicating that the ORF58 gene is non-essential for virus growth. Comparison of the growth rate of rpOkaΔ58 and recombinant wild-type virus by assessing plaque sizes revealed no significant differences between them both in MRC-5 cells and malignant melanoma cells.

Conclusion

This study shows that the ORF58 gene is dispensable for viral replication and does not affect the virus' ability to form plaques in vitro.
A values-based perspective on good practice in
compulsion
Authors: Fulford K. W. M. , King Malcolm
Published: April 2008

We outline how the values-based approach adopted in training materials
supporting the Mental Health Act 2007 for England and Wales will complement
recovery-based practice in compulsory psychiatric detention.
Post-traumatic stress disorder in serious accidental injury: 3-year follow-up study
Authors: Hepp Urs , Moergeli Hanspeter , Buchi Stefan , Bruchhaus-Steinert Helke , Kraemer Bernd , Sensky Tom , Schnyder Ulrich
Published: April 2008

Background

Long-term data on post-traumatic stress disorder (PTSD) following accidents are scarce.

Aims

To assess and predict PTSD in people 3 years after severe accidental injury.

Method

Severely injured patients were recruited consecutively from the intensive care unit (n=121) and assessed within 1 month of the trauma. Follow-up interviews were conducted 6 months, 12 months and 36 months later; 90 patients participated in all four interviews. Symptoms were assessed using the Clinician-Administered PTSD Scale.

Results

Post-traumatic stress disorder was diagnosed in 6% of patients 2 weeks after the accident, in 2% after 1 year and in 4% after 3 years. Robust predictors of later PTSD symptom level were intrusive symptoms shortly after the accident and biographical risk factors. There were individual changes over time between the categories PTSD, sub-threshold PTSD and no PTSD. Whereas PTSD symptom severity was low or decreased for most of the patients, some of them showed an increase or a delayed onset. Patients with persisting PTSD symptoms at 6 months and patients with delayed onset of symptoms are at risk of long-term PTSD.

Conclusions

The prevalence of PTSD was low over the whole period of 3 years.
Duration of antidepressant use during pregnancy and risk of
major congenital malformations
Authors: Ramos Élodie , St-André Martin , Rey Évelyne , Oraichi Driss , Bérard Anick
Published: April 2008

Background

Antidepressant use during the gestational period is a controversial
topic.

Aims

To determine whether duration of antidepressant use during the first
trimester increases the risk of major congenital malformations in
offspring of women diagnosed with psychiatric disorders.

Method

A case-control study was performed among women who had been pregnant
between January 1998 and December 2002. Data were obtained from a
Medication and Pregnancy registry, built by linking three databases from
the province of Quebec, and a self-administered questionnaire. Women
eligible for this study had to be 15–45 years old at the beginning of
pregnancy, have at least one diagnosis of psychiatric disorder before
pregnancy, have used antidepressants for ≥ 30 days in the year prior to
pregnancy and have a pregnancy ending with a delivery. Cases were defined
as any major congenital malformation diagnosed in the offspring's first
year of life. Odds ratios, adjusted for relevant confounders, were
estimated using logistic regression.

Results

Among the 2329 women meeting the inclusion criteria, 189 (8.1%) infants
were born with a major congenital malformation. Duration of
antidepressant use during the first trimester of pregnancy was not
associated with an increased risk of major congenital malformations: 1–30
days v. 0 day, adjusted OR=1.23 (95% CI 0.77–1.98);
31–60 days v. 0 day, adjusted OR=1.03 (95% CI
0.63–1.69); ≥ 61 days v. 0 day, adjusted OR=0.92 (95% CI
0.50–1.69).

Conclusions

These data do not support an association between duration of
antidepressant use during the first trimester of pregnancy and major
congenital malformations in the offspring of women with psychiatric
disorders. These findings should help clinicians decide whether to
continue antidepressant therapy during pregnancy.
Affective modulation of anterior cingulate cortex in young
people at increased familial risk of depression
Authors: Mannie Zola N. , Norbury Ray , Murphy Susannah E. , Inkster Becky , Harmer Catherine J. , Cowen Philip J.
Published: April 2008

Background

We previously found that children of parents with depression showed
impaired performance on a task of emotional categorisation.

Aims

To test the hypothesis that children of parents with depression would
show abnormal neural responses in the anterior cingulate cortex, a brain
region involved in the integration of emotional and cognitive
information.

Method

Eighteen young people (mean age 19.8 years) with no personal history of
depression but with a biological parent with a history of major
depression (FH+ participants) and 16 controls (mean age 19.9
years) underwent functional magnetic resonance imaging while completing
an emotional counting Stroop task.

Results

Controls showed significant activation in the pregenual anterior
cingulate cortex to both positive and negative words during the emotional
Stroop task. This activation was absent in FH+
participants.

Conclusions

Our findings show that people at increased familial risk of depression
demonstrate impaired modulation of the anterior cingulate cortex in
response to emotionally valenced stimuli.
Clinical differences between bipolar and unipolar
depression
Authors: Forty Liz , Smith Daniel , Jones Lisa , Jones Ian , Caesar Sian , Cooper Carly , Fraser Christine , Gordon-Smith Katherine , Hyde Sally , Farmer Anne , McGuffin Peter , Craddock Nick
Published: April 2008

It is commonly – but wrongly – assumed that there are no important
differences between the clinical presentations of major depressive disorder
and bipolar depression. Here we compare clinical course variables and
depressive symptom profiles in a large sample of individuals with major
depressive disorder (n=593) and bipolar disorder
(n=443). Clinical characteristics associated with a
bipolar course included the presence of psychosis, diurnal mood variation
and hypersomnia during depressive episodes, and a greater number of shorter
depressive episodes. Such features should alert a clinician to a possible
bipolar course. This is important because optimal management is not the same
for bipolar and unipolar depression.
Dr Frances Felicity Kerr (née Gibbs)
Authors: Rollin Henry R.
Published: April 2008

Felicity Kerr (her preferred name) was born on 3rd June 1923 into an affluent family in South Wales. It was traditional, within her social class and at that time, to send children to boarding school: in Felicity's case, to West Heath School in Kent. Unfortunately, this coincided with the perils of World War II and the onslaught of the Luftwaffe which, too frequently, chose to cross the coast dangerously near the school. In the interests of her safety, Felicity was moved back to Wales where, at Cardiff Technical College, she matriculated.
Nɛ‐(Carboxymethyl)lysine during the Early Development of Hypertension
Authors: Baumann Marcus , Stehouwer Coen , Scheijen Jean , Heemann Uwe , Struijker Boudier Harry , Schalkwijk Casper
Published: April 2008

Advanced glycation end products (AGEs) are associated with hypertension. Whether Nɛ‐(carboxymethyl)lysine (CML) contributes to the development of hypertension in young spontaneously hypertensive rats (SHR) remains to be established compared to WKY. We determined blood pressure, renal function, marker for oxidative stress (OS), and CML in young WKY rats and SHR. We found blood pressure was increased in SHR with no difference in renal function and OS compared to WKY. CML was elevated in plasma (2.3 ± 0.3 vs. 1.3 ± 0.2 μmol/L) and kidney (1.0 ± 0.1 vs. 0.5 ± 0.1 μmol/L) compared to WKY. Early CML accumulation may contribute to the development of hypertension potentially by inducing early renal inflammation independent of glomerular dysfunction or oxidative stress.
The Aroma Side of the Maillard Reaction
Authors: Cerny Christoph
Published: April 2008

The Maillard reaction in food produces, among others, a diversity of sensory‐active compounds (aroma, taste, color). The resulting key aroma compounds are often present only in trace concentrations of 1 μg/kg to 1 mg/kg. Nevertheless, they contribute to the respective flavor because of their low odor‐perception thresholds. While Maillard intermediates, such as Amadori compounds and deoxyosones, are formed at percentage levels during model reactions, the yield of aroma compounds, in particular nitrogen and sulfur‐containing ones, is often as low as 0.001–0.01 mol%, thus indicating their formation through chemical side reactions. The elucidation of the relevant precursors in food and the identification of previously unknown intermediates can throw light on these minor pathways. Also, model reactions with isotopically labeled precursors are of great value in gaining insight into the relevant formation mechanisms. Several examples of these studies are illustrated including work to elucidate the role of the solvent glycerol in the formation of pyrazines, trials to reveal the relative significance of 4‐hydroxy‐5‐methyl‐3(2H)‐furanone as intermediate in the reaction between ribose and cysteine, and experiments to assess the proportional contribution of the precursors cysteine, xylose, and thiamine to the formation of the resulting aroma compounds in the thermal reaction.
Evaluating the Extent of Protein Damage in Dairy Products
Authors: Hegele Jörg , Parisod Véronique , Richoz Janique , Förster Anke , Maurer Sarah , Krause René , Henle Thomas , Bütler Timo , Delatour Thierry
Published: April 2008

An isotope dilution liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method was developed to determine lysine (Lys), Nε‐fructosyllysine (FL), Nε‐carboxymethyllysine (CML), and pyrraline (Pyr) in dairy products. The presented approach entails protein cleavage via enzymatic digestion to liberate the aforementioned compounds, which were then quantified using a stable isotope dilution assay. LC‐MS/MS analysis was performed by positive electrospray ionization recording two transition reactions per analyte in selected reaction monitoring mode. The CML and Lys values obtained with enzymatic digestion were compared to those acquired with acid hydrolysis HCl (6 mol/L), and the two proteolysis methods yielded comparable quantifications. Allowing for the fact that the investigated compounds are formed during different stages of the glycation process, the method is able to reveal the progress of protein glycation in dairy products.
Application of Mass Spectrometry for the Detection of Glycation and Oxidation Products in Milk Proteins
Authors: Meltretter Jasmin , Pischetsrieder Monika
Published: April 2008

Protein mass spectometry techniques, such as electrospray ionization mass spectrometry or matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS), are effective methods to screen for protein modifications derived from the Maillard reaction. The analysis of the intact proteins reveals the major modification, most commonly the Amadori product, whereas partial enzymatic hydrolysis prior to mass spectrometry additionally allows the detection of minor adducts. Therefore, a mass spectrometric method was developed for the analysis of whey protein modifications occurring during heat treatment. The two main whey proteins, α‐lactalbumin and β‐lactoglobulin, were incubated with lactose in a milk model and modifications were recorded using MALDI‐TOF‐MS. The analysis of the intact proteins revealed protein species with 0–4 lactulosyl residues. Partial enzymatic hydrolysis with endoproteinase AspN prior to mass spectrometric analysis enabled the detection of further modifications and their localization in the amino acid sequence. Detected modifications were lactulosyllysine, Nε‐(carboxymethyl)lysine, lysine aldehyde, methionine sulfoxide, cyclization of N‐terminal glutamic acid to a pyrrolidone, and oxidation of cysteine or tryptophan. Protein modifications in heated milk and commercially available dairy products can be analyzed after the separation of the milk proteins using one‐dimensional SDS‐PAGE.
Acrolein Induces Inflammatory Response Underlying Endothelial Dysfunction
Authors: Park Yong Seek , Taniguchi Naoyuki
Published: April 2008

Endothelial dysfunction by proinflammatory stimuli represents an important link between risk factors and the pathologic mechanisms underlying atherosclerosis. Thus, control of the inflammatory status of endothelial cells is crucial to limiting the disease. Tobacco smoking induces inflammatory reactions and promotes atherosclerosis; however, the mechanism that links cigarette smoking to an increased incidence of atherosclerosis is poorly understood. Our study demonstrates that acrolein, a known toxin in tobacco smoke, elevates oxidative stress via inactivation of thioredoxin reductase and stimulates expression of cyclooxygenase‐2 through activation of the protein kinase C, p38 mitogen‐activated protein kinase, and cAMP response element‐binding protein pathway in endothelial cells. Our finding suggests that acrolein may play a role in the progression of atherosclerosis.
Preparation of Nucleotide Advanced Glycation Endproducts—Imidazopurinone Adducts Formed by Glycation of Deoxyguanosine with Glyoxal and Methylglyoxal
Authors: Fleming Thomas , Rabbani Naila , Thornalley Paul J.
Published: April 2008

An analytical procedure was developed for nucleotide advanced glycation endproducts formed by the reaction of glyoxal and methylglyoxal with deoxyguanosine under physiological conditions. For this, the imidazopurinone derivatives, 3‐(2′‐deoxyribosyl)‐6,7‐dihydro‐6,7‐dihydroxyimidazo[2,3‐b]purin‐9(8)one (dG‐G) and 3‐(2′‐deoxyribosyl)‐6,7‐dihydro‐6,7‐dihydroxy‐6‐methylimidazo‐[2,3‐b]purine‐9(8)one (dG‐MG), were prepared. Authentic standard and stable isotope‐substituted standard adducts were prepared and an isotopic dilution analysis assay methodology was developed using liquid chromatography with tandem mass spectrometry and optimized DNA extraction and nuclease digestion procedures. Analysis of dG‐G, dG‐MG, and the oxidative marker 8‐hydroxydeoxyguanosine in the DNA of cultured human cells and mononuclear leukocytes showed that nucleotide advanced glycation endproducts are major markers of DNA damage in human cells.
Advanced Glycation Endproducts in Chronic Heart Failure
Authors: Smit Andries J. , Hartog Jasper W. L. , Voors Adriaan A. , Van Veldhuisen Dirk J.
Published: April 2008

Advanced glycation endproducts (AGEs) have been proposed as factors involved in the development and progression of chronic heart failure (CHF). Cross‐linking by AGEs results in vascular and myocardial stiffening, which are hallmarks in the pathogenesis of CHF. Additionally, stimulation of receptors by AGEs may affect endothelial function and myocardial calcium uptake and may perpetuate coronary sclerosis in CHF. CHF is common in conditions with AGE accumulation, such as diabetes and renal failure. This review describes in detail the interrelation of plasma AGEs, renal function, and the severity and prognosis in clinical CHF patients with mild to moderate loss of renal function. This association is compared with the relation between tissue AGE accumulation (marked by skin autofluorescence) and diastolic dysfunction in renal failure. The evidence reviewed here provides support for the assumed role of AGEs in determining the severity and prognosis of CHF, but also highlights the differences in this relation between plasma and tissue AGEs and between patients with and without advanced renal failure. Ongoing clinical intervention trials to reduce AGE accumulation in patients with CHF may elucidate the causal role of AGEs in the development and course of CHF.
Modification of Vimentin
Authors: Kueper Thomas , Grune Tilman , Muhr Gesa‐Meike , Lenz Holger , Wittern Klaus‐Peter , Wenck Horst , Stäb Franz , Blatt Thomas
Published: April 2008

In a recent study, we were able to show that the intermediate filament protein vimentin aggregates in human dermal fibroblasts because of modification by the advanced glycation endproduct carboxymethyllysine (CML). In this work, we investigated the formation of intracellular CML in relation to the concentration of glucose in the culture medium. The natural degradation product of glucose, methylglyoxal, was able to induce the aggregation of vimentin. This dicarbonyl leads to the formation of the modifications MG‐H1 and carboxyethyllysine (CEL) as a result of the reaction with arginine and lysine residues of proteins. Furthermore, we found that the protein vimentin was modified, not only by CML and CEL, but also by pentosidine and pyrraline. These findings underline the special position of vimentin as a preferential target of the Maillard reaction in human skin.
A Novel Yellow Pigment, Furpipate, Derived from Lysine and Furfural
Authors: Murata Masatsune , Totsuka Hana , Ono Hiroshi
Published: April 2008

Furfural is an important intermediate compound of the Maillard reaction of pentose or ascorbic acid. We examined the browning of furfural and lysine by heating these compounds in solution and found two yellow compounds. The first, furpipate, is a novel pipecolic acid derivative and was (Z)‐3‐(2‐furylmethylidene)‐3H, 4H, 5H, 6H‐pyridine‐2‐carboxylic acid. The second was decarboxylated‐furpipate. Furpipate shows absorption maxima at 370 nm and 310 nm under acidic and alkaline conditions, respectively. This compound was the major colored compound of the heated solution containing lysine and furfural.
Receptor for Advanced Glycation End Products
Authors: Ramasamy Ravichandran , Yan Shi Fang , Herold Kevan , Clynes Raphael , Schmidt Ann Marie
Published: April 2008

The multiligand receptor for advanced glycation end products (RAGE) of the immunoglobulin superfamily is expressed on multiple cell types implicated in the immune–inflammatory response and in atherosclerosis. Multiple studies have elucidated that ligand–RAGE interaction on cells, such as monocytes, macrophages, and endothelial cells, mediates cellular migration and upregulation of proinflammatory and prothrombotic molecules. In addition, recent studies reveal definitive rules for RAGE in effective T lymphocyte priming in vivo. RAGE ligand AGEs may be formed in diverse settings; although AGEs are especially generated in hyperglycemia, their production in settings characterized by oxidative stress and inflammation suggests that these species, in part via RAGE, may contribute to the pathogenesis of atherosclerosis. In murine models of atherosclerosis, vascular inflammation is a key factor and one which is augmented, in parallel with even further increases in RAGE ligands, in diabetic macrovessels. The findings that antagonism and genetic disruption of RAGE in atherosclerosis‐susceptible mice strikingly reduces vascular inflammation and atherosclerotic lesion area and complexity link RAGE intimately to these processes and suggest that RAGE is a logical target for therapeutic intervention in aberrant inflammatory mechanisms and in atherosclerosis.
α‐Dicarbonyl Compounds—Key Intermediates for the Formation of Carbohydrate‐based Melanoidins
Authors: Kroh Lothar W. , Fiedler Thorsten , Wagner Janine
Published: April 2008

The Maillard reaction of carbohydrates and amino acids is the chemical basis for flavor and color formation in many processed foods. Dicarbonyl compounds, such as 1‐, 3‐deoxyosones and 1,4‐dideoxyosones, as well as short‐chain dicarbonyls, such as methylgyoxal or glyoxal, are key compounds of the Maillard browning reaction. The α‐dicarbonyls are also starting materials for polymerization reactions which lead to formation of carbohydrate‐based melanoidins. With regard to the dicarbonyl compound, different possible chemical structures of melanoidins will be discussed. The analysis by size‐exclusion chromatography revealed that those colored compounds differ in their molecular size and are directly associated with reactions having specific α‐dicarbonyl compounds.
Time‐dependent Component‐specific Regulation of Gastric Acid Secretion‐related Proteins by Roasted Coffee Constituents
Authors: Rubach M. , Lang R. , Hofmann T. , Somoza V.
Published: April 2008

Consumption of coffee beverages has been reported to cause gastric irritation in some consumers as a result of increased gastric acid secretion. In the complex mechanisms of gastric acid secretion, the activity and expression of the H+,K+‐ATPase is regulated by transmitters, such as histamine, acetylcholine, gastrin, somatostatin, and their corresponding receptors. Here, we report the effect of three coffee constituents, chlorogenic acid, caffeine, and N‐methyl pyridinium ions, on the expression of the histamine receptor H2, the acetylcholine receptor M3, the gastrin receptor, the somatostatin receptor, and the H+,K+‐ATPase. Human gastric cancer cells were exposed to chlorogenic acid, caffeine, or N‐methyl pyridinium in their coffee brew‐representative concentrations as well as to physiological stimulators of gastric acid secretion. Gene expression levels of receptor proteins and those of the H+,K+‐ATPase were measured at different time points by real‐time PCR. Expression of prosecretory receptors significantly increased between one and one‐half to twofold after treatment with chlorogenic acid or caffeine compared to control cells at the same time point. Chlorogenic acid and caffeine also increased the H+,K+‐ATPase gene expression twofold higher compared to control cells. In contrast, N‐methyl pyridinium downregulated the expression of the prosecretory gastrin receptor significantly, by −27%. In conclusion, chlorogenic acid, caffeine, and N‐methyl pyridinium impair the expression of gastric acid secretion‐related proteins in a time‐dependent manner. Future work will be aimed at the elucidation of the cooperative interplay of individual components using recombinates of single coffee constituents.
Long-Term Cardiovascular Effects of Levothyroxine Therapy in Young Adults with Congenital Hypothyroidism
Authors: Salerno Mariacarolina , Oliviero Ugo , Lettiero Teresa , Guardasole Vincenzo , Mattiacci Dario Maria , Saldamarco Lavinia , Capalbo Donatella , Lucariello Antonio , Saccà Luigi , Cittadini Antonio
Published: April 2008

Context: Congenital hypothyroidism (CH) is the most prevalent endocrine disorder in the newborn and is routinely treated with life-long levothyroxine replacement therapy. Although several studies have demonstrated that such therapy may impact on the cardiovascular system, little is known with regard to the effects of long-term levothyroxine administration in patients with CH.

Objective: The aim of the current study was to evaluate whether long-term levothyroxine replacement therapy in young adults with CH is associated with cardiovascular abnormalities.

Patients and Methods: Thirty young adults with CH aged 18.1 ± 0.2 yr and 30 age- and sex-matched controls underwent cardiac and carotid Doppler ultrasound and symptom-limited cardiopulmonary exercise testing. Hypothyroidism was diagnosed by neonatal screening, and levothyroxine treatment was initiated within the first month of life and carefully adjusted to maintain TSH levels in the normal range and free T4 in the high-normal range.

Results: Compared with controls, hypothyroid patients exhibited left ventricular diastolic dysfunction, impaired exercise capacity, and increased intima-media thickness. At multiple regression analysis, the number of episodes of plasma TSH levels less than 0.5 mU/liter and greater than 4.0 mU/liter from the age of 1 yr onward, and mean TSH plasma levels during puberty were independent predictors of diastolic filling and cardiopulmonary performance indexes (multiple r values: 0.61–0.75).

Conclusions: Long-term levothyroxine treatment in young adults with congenital hypothyroidism is associated with impaired diastolic function and exercise capacity and increased intima-media thickness.
Longitudinal Study of Thyroid Function in Children with Mild Hyperthyrotropinemia at Neonatal Screening for Congenital Hypothyroidism
Authors: Leonardi Daniela , Polizzotti Nunziella , Carta Anna , Gelsomino Rossella , Sava Lidia , Vigneri Riccardo , Calaciura Francesca
Published: April 2008

Objective: Long-term outcome of thyroid function in children with very short-lasting neonatal hyperthyrotropinemia (“false positive” at neonatal screening) was studied in an observational, prospective study. Thyroid function and morphology were evaluated in 44 “false positive” children up to advanced childhood (8.0 ± 0.7 yr of age). In these children a high prevalence (50%) of subclinical hypothyroidism in early childhood (2.8 ± 0.5 yr) had already been described.

Results: At an average of 5.3 yr, subclinical hypothyroidism persisted in 19 of 44 (43.2%) children and, more specifically, in two of three of those who had increased TSH in early childhood. Euthyroidism was present in all cases that were euthyroid in early childhood, although they had TSH and free T3 values significantly higher than control children with a normal TSH at birth (TSH = 2.6 ± 0.7 vs. 1.5 ± 0.6 mU/liter, P < 0.001; free T3 = 4.9 ± 0.8 vs. 3.9 ± 0.9 pmol/liter, P < 0.01). Thyroid morphology alterations were frequent in the group of children with subclinical hypothyroidism. At an average of 8.0 yr, subclinical hypothyroidism persisted in 14 of 44 (31.8%) children. In all other children, TSH and thyroid hormones were confirmed within the normal range.

Conclusions: This prospective longitudinal study confirms that newborns “false positive” at neonatal screening have a high risk to develop persistent subclinical hypothyroidism. The prevalence of hypothyroidism decreases with increasing age, but it is still high (>30%) in late childhood. Even those “false positive” children that maintain euthyroidism in late childhood have an average TSH value that, although within the normal range, is higher than in normal controls, a possible marker of minor congenital thyroid function abnormalities.
Leptin Does Not Mediate Short-Term Fasting-Induced Changes in Growth Hormone Pulsatility but Increases IGF-I in Leptin Deficiency States
Authors: Chan Jean L. , Williams Catherine J. , Raciti Patricia , Blakeman Jennifer , Kelesidis Theodore , Kelesidis Iosif , Johnson Michael L. , Thorner Michael O. , Mantzoros Christos S.
Published: April 2008

Context: States of acute and chronic energy deficit are characterized by increased GH secretion and decreased IGF-I levels.

Objective: The objective of the study was to determine whether changes in levels of leptin, a key mediator of the adaptation to starvation, regulate the GH-IGF system during energy deficit.

Design, Setting, Patients, and Intervention: We studied 14 healthy normal-weight men and women during three conditions: baseline fed and 72-h fasting (to induce hypoleptinemia) with administration of placebo or recombinant methionyl human leptin (r-metHuLeptin) (to reverse the fasting associated hypoleptinemia). We also studied eight normal-weight women with exercise-induced chronic energy deficit and hypothalamic amenorrhea at baseline and during 2–3 months of r-metHuLeptin treatment.

Main Outcome Measures: GH pulsatility, IGF levels, IGF and GH binding protein (GHBP) levels were measured.

Results: During short-term energy deficit, measures of GH pulsatility and disorderliness and levels of IGF binding protein (IGFBP)-1 increased, whereas leptin, insulin, IGF-I (total and free), IGFBP-4, IGFBP-6, and GHBP decreased; r-metHuLeptin administration blunted the starvation-associated decrease of IGF-I. In chronic energy deficit, total and free IGF-I, IGFBP-6, and GHBP levels were lower, compared with euleptinemic controls; r-metHuLeptin administration had no major effect on GH pulsatility after 2 wk but increased total IGF-I levels and tended to increase free IGF-I and IGFBP-3 after 1 month.

Conclusions: The GH/IGF system changes associated with energy deficit are largely independent of leptin deficiency. During acute energy deficit, r-metHuLeptin administration in replacement doses blunts the starvation-induced decrease of IGF-I, but during chronic energy deficit, r-metHuLeptin administration increases IGF-I and tends to increase free IGF-I and IGFBP-3.
Association of CYP3A7*1C and Serum Dehydroepiandrosterone Sulfate Levels in Women with Polycystic Ovary Syndrome
Authors: Goodarzi Mark O. , Xu Ning , Azziz Ricardo
Published: April 2008

Context: Adrenal androgen excess is common in polycystic ovary syndrome (PCOS) and appears to be heritable. CYP3A7 metabolizes dehydroepiandrosterone and its sulfate (DHEAS). A promoter variant, CYP3A7*1C, which results in persistent expression in adults, was associated with reduced DHEAS levels in a previous study, which led us to consider CYP3A7*1C as a modulator of adrenal androgen excess in patients with PCOS.

Objective: The objective was to replicate the association between CYP3A7*1C and reduced DHEAS levels in PCOS patients and assess its possible role in modulating testosterone levels.

Design: Women with and without PCOS were genotyped for CYP3A7*1C, and this variant was tested for association with DHEAS and total and free testosterone.

Setting: Subjects were recruited from the reproductive endocrinology clinic at the University of Alabama at Birmingham; controls were recruited from the surrounding community. Genotyping took place at Cedars-Sinai Medical Center (Los Angeles, CA).

Participants: A total of 287 white women with PCOS and 187 controls were studied.

Main Measurements: CYP3A7*1C genotype, PCOS risk, and androgen levels were measured.

Results: PCOS subjects who carried the CYP3A7*1C variant had lower levels of serum DHEAS and total testosterone (P = 0.0006 and 0.046, respectively). The variant was not associated with PCOS risk.

Conclusion: This study replicated prior work of the association of CYP3A7*1C and decreased DHEAS in a different population of young PCOS women, providing further genetic evidence that CYP3A7 plays a potential role in modulation of DHEAS levels. Adult expression of CYP3A7 may modify the PCOS phenotype by ameliorating adrenal androgen excess.
Integrin clustering enables anandamide-induced Ca2+ signaling in endothelial cells via GPR55 by protection against CB1-receptor-triggered repression
Authors: Waldeck-Weiermair Markus , Zoratti Cristina , Osibow Karin , Balenga Nariman , Goessnitzer Edith , Waldhoer Maria , Malli Roland , Graier Wolfgang F.
Published: April 2008

Although the endocannabinoid anandamide is frequently described to act predominantly in the cardiovascular system, the molecular mechanisms of its signaling remained unclear. In human endothelial cells, two receptors for anandamide were found, which were characterized as cannabinoid 1 receptor (CB1R; CNR1) and G-protein-coupled receptor 55 (GPR55). Both receptors trigger distinct signaling pathways. It crucially depends on the activation status of integrins which signaling cascade becomes promoted upon anandamide stimulation. Under conditions of inactive integrins, anandamide initiates CB1R-derived signaling, including Gi-protein-mediated activation of spleen tyrosine kinase (Syk), resulting in NFκB translocation. Furthermore, Syk inhibits phosphoinositide 3-kinase (PI3K) that represents a key protein in the transduction of GPR55-originated signaling. However, once integrins are clustered, CB1R splits from integrins and, thus, Syk cannot further inhibit GPR55-triggered signaling resulting in intracellular Ca2+ mobilization from the endoplasmic reticulum (ER) via a PI3K-Bmx-phospholipase C (PLC) pathway and activation of nuclear factor of activated T-cells. Altogether, these data demonstrate that the physiological effects of anandamide on endothelial cells depend on the status of integrin clustering.
Haemocyte-derived SPARC is required for collagen-IV-dependent stability of basal laminae inDrosophilaembryos
Authors: Martinek Nathalie , Shahab Jaffer , Saathoff Manuela , Ringuette Maurice
Published: April 2008

SPARC is an evolutionarily conserved collagen-binding extracellular matrix (ECM) glycoprotein whose morphogenetic contribution(s) to embryonic development remain elusive despite decades of research. We have therefore used Drosophila genetics to gain insight into the role of SPARC during embryogenesis. In Drosophila embryos, high levels of SPARC and other basal lamina components (such as network-forming collagen IV, laminin and perlecan) are synthesized and secreted by haemocytes, and assembled into basal laminae. A SPARC mutant was generated by P-element mutagenesis that is embryonic lethal because of multiple developmental defects. Whereas no differences in collagen IV immunostaining were observed in haemocytes between wild-type and SPARC-mutant embryos, collagen IV was not visible in basal laminae of SPARC-mutant embryos. In addition, the laminin network of SPARC-mutant embryos appeared fragmented and discontinuous by late embryogenesis. Transgenic expression of SPARC protein by haemocytes in SPARC-mutant embryos restored collagen IV and laminin continuity in basal laminae. However, transgenic expression of SPARC by neural cells failed to rescue collagen IV in basal laminae, indicating that the presence of collagen IV deposition requires SPARC expression by haemocytes. Our previous finding that haemocyte-derived SPARC protein levels are reduced in collagen-IV-mutant embryos and the observation that collagen-IV-mutant embryos showed a striking phenotypic similarity to SPARC-mutant embryos suggests a mutual dependence between these major basal laminae components during embryogenesis. Patterning defects and impaired condensation of the ventral nerve cord also resulted from the loss SPARC expression prior to haemocyte migration. Hence, SPARC is required for basal lamina maturation and condensation of the ventral nerve cord during Drosophila embryogenesis.
Ubiquitin-independent binding of Hrs mediates endosomal sorting of the interleukin-2 receptor β-chain
Authors: Yamashita Yuki , Kojima Katsuhiko , Tsukahara Tomonori , Agawa Hideyuki , Yamada Koichiro , Amano Yuji , Kurotori Naoki , Tanaka Nobuyuki , Sugamura Kazuo , Takeshita Toshikazu
Published: April 2008

Several lines of evidence have revealed that ubiquitylation of membrane proteins serves as a signal for endosomal sorting into lysosomes or lytic vacuoles. The hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs) interacts with ubiquitylated cargoes through its ubiquitin-interacting-motif domain (UIM domain), and plays an essential early role in endosomal sorting. Here, we show that the C-terminal region of Hrs, which does not contain the UIM domain, can bind to interleukin-2 receptor β (IL-2Rβ). We found a direct interaction between bacterially expressed IL-2Rβ and Hrs in GST pull-down assays, indicating that their binding is independent of ubiquitin. Trafficking and degradation assays revealed that, similarly to wild-type IL-2Rβ, an IL-2Rβ mutant lacking all the cytoplasmic lysine residues is sorted from Hrs-positive early endosomes to LAMP1-positive late endosomes, resulting in degradation of the receptor. By contrast, an IL-2Rβ mutant lacking the Hrs-binding region passes through early endosomes and is mis-sorted to compartments positive for the transferrin receptor. The latter mutant exhibits attenuated degradation. Taken together, these results indicate that precise sorting of IL-2Rβ from early to late endosomes is mediated by Hrs, a known sorting component of the ubiquitin-dependent machinery, in a manner that is independent of UIM-ubiquitin binding.
Dnm1p-dependent peroxisome fission requires Caf4p, Mdv1p and Fis1p
Authors: Motley Alison M. , Ward Gemma P. , Hettema Ewald H.
Published: April 2008

Yeast peroxisomes multiply by fission. Fission requires two dynamin-related proteins, Dnm1p and Vps1p. Using an in vivo fission assay, we show that Dnm1p-dependent peroxisome fission requires Fis1p, Caf4p and Mdv1p. Fluorescence microscopy of cells expressing GFP-tagged Caf4p and Mdv1p revealed that their association with peroxisomes relies on Fis1p. Vps1p-dependent peroxisome fission occurs independently of these factors. Vps1p contributes most to fission of peroxisomes when cells are grown on glucose. Overexpression of Dnm1p suppresses the fission defect as long as Fis1p and either Mdv1p or Caf4p are present. Conversely, overexpression of Dnm1p does not restore the vacuolar fusion defect of vps1 cells and Vps1p overexpression does not restore the mitochondrial fission defect of dnm1 cells. These data show that Vps1p and Dnm1p are part of independent fission machineries. Because the contribution of Dnm1p to peroxisome fission appears to be more pronounced in cells that proliferate peroxisomes in response to mitochondrial dysfunction, Dnm1p might be part of the mechanism that coordinates mitochondrial and peroxisomal biogenesis.
Inter- and intra-observer variability of the Crowe and Hartofilakidis classification systems for congenital hip disease in adults
Authors: Yiannakopoulos C. K. , Chougle A. , Eskelinen A. , Hodgkinson J. P. , Hartofilakidis G.
Published: April 2008

Our study evaluated the reliability of the Crowe and Hartofilakidis classification systems for developmental dysplasia of the hip in adults. The anteroposterior radiographs of the pelvis of 145 patients with 209 osteoarthritic hips were examined twice by three experienced hip surgeons from three European countries and the abnormal hips were rated using both classifications. The inter- and intra-observer agreement was calculated.

Interobserver reliability was evaluated using weighted and unweighted kappa coefficients and for the Crowe classification, among the three pairs there was a minimum kappa coefficient with linear weighting of 0.90 for observers A and C and a maximum kappa coefficient of 0.92 for observers B and C. For the Hartofilakidis classification, the minimum kappa value was 0.85 for observers A and B, and the maximum value was 0.93 for observers B and C. With regard to intra-observer reliability, the kappa coefficients with linear weighting between the two evaluations of the same observer ranged between 0.86 and 0.95 for the Crowe classification and between 0.80 and 0.93 for the Hartofilakidis classification.

The reliability of both systems was substantial to almost perfect both for serial measurements by individual readers and between different readers, although the information offered was dissimilar.