bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid detection of trisomy 21 by gene dosage analysis using quantitative real‐time polymerase chain reaction
Authors: Tsujie Tomoko , Takemura Masahiko , Kimura Tadashi , Shimoya Koichiro , Tsutsui Tateki , Ogita Kazuhide , Ozaki Mamoru , Murata Yuji
Published: July 2006

Aim:  Rapid detection of fetal aneuploidy helps inform a mother’s choice about the course of her pregnancy. Obtaining results by fluorescent in situ hybridization (FISH) requires more than 24 h, and thus a more rapid method is needed.

Methods:  Conventional G‐banding and FISH for chromosome 21 were performed for cultured amniocytes. Genomic DNA was extracted from uncultured amniocytes obtained from 23 patients. TaqMan polymerase chain reaction (PCR) primers were designed to amplify the potassium voltage gated channel gene on chromosome 21q22.12 and the ribosomal phosphoprotein gene on 18q21.1. Quantitative real‐time PCR was performed for these two gene fragments and the differences of the threshold cycle (Ct) of the two genes (Ct 18–Ct 21) were calculated for each sample.

Results:  G‐banding revealed that 19 patients had a normal karyotype and four had trisomy 21. FISH resulted in one case of a false positive. The ΔCt values (Ct 18–Ct 21) of trisomy 21 patients were significantly higher than the values of individuals with normal karyotypes (P < 0.001) and there was no overlapping.

Conclusions:  Fetal trisomy 21 is rapidly detectable by gene dosage analysis from amniocytes using quantitative real‐time PCR.
Microarray and genetic analysis of electron transfer to electrodes in Geobacter sulfurreducens
Authors: Holmes Dawn E. , Chaudhuri Swades K. , Nevin Kelly P. , Mehta Teena , Methé Barbara A. , Liu Anna , Ward Joy E. , Woodard Trevor L. , Webster Jennifer , Lovley Derek R.
Published: July 2006

Whole‐genome analysis of gene expression in Geobacter sulfurreducens revealed 474 genes with transcript levels that were significantly different during growth with an electrode as the sole electron acceptor versus growth on Fe(III) citrate. The greatest response was a more than 19‐fold increase in transcript levels for omcS, which encodes an outer‐membrane cytochrome previously shown to be required for Fe(III) oxide reduction. Quantitative reverse transcription polymerase chain reaction and Northern analyses confirmed the higher levels of omcS transcripts, which increased as power production increased. Deletion of omcS inhibited current production that was restored when omcS was expressed in trans. Transcript expression and genetic analysis suggested that OmcE, another outer‐membrane cytochrome, is also involved in electron transfer to electrodes. Surprisingly, genes for other proteins known to be important in Fe(III) reduction such as the outer‐membrane c‐type cytochrome, OmcB, and the electrically conductive pilin ‘nanowires’ did not have higher transcript levels on electrodes, and deletion of the relevant genes did not inhibit power production. Changes in the transcriptome suggested that cells growing on electrodes were subjected to less oxidative stress than cells growing on Fe(III) citrate and that a number of genes annotated as encoding metal efflux proteins or proteins of unknown function may be important for growth on electrodes. These results demonstrate for the first time that it is possible to evaluate gene expression, and hence the metabolic state, of microorganisms growing on electrodes on a genome‐wide basis and suggest that OmcS, and to a lesser extent OmcE, are important in electron transfer to electrodes. This has important implications for the design of electrode materials and the genetic engineering of microorganisms to improve the function of microbial fuel cells.
Transcription and mass‐spectroscopic proteomic studies of electron transport oxidoreductases in Dehalococcoides ethenogenes
Authors: Morris R. M. , Sowell S. , Barofsky D. , Zinder S. , Richardson R.
Published: July 2006

Besides 19 potential reductive dehalogenase genes, the genome of Dehalococcoides ethenogenes strain 195 contains over 60 genes annotated as encoding oxidoreductases, including five hydrogenase complexes and a formate dehydrogenase (Fdh). Using quantitative reverse transcriptase polymerase chain reaction, we found that genes encoding a periplasmic Hup hydrogenase and the Fdh were the most highly expressed in batch‐grown pure cultures, in which the H2 partial pressure was > 0.1 atm, and in butyrate/tetrachloroethene‐mixed cultures, in which H2 partial pressures were 10−4−10−5 atm. Shotgun electrospray ionization (ESI) and matrix‐assisted laser desorption/ionization (MALDI) tandem mass spectrometry were used to identify multiple peptides in pure culture membrane‐enriched fractions matching several highly expressed respiratory enzymes, including three hydrogenases, two reductive dehalogenases, Fdh and DET1407, a 105.5‐kDa protein we propose to be part of an S‐layer cell wall. Both transcript and mass spectrometric approaches indicated that the putative Fdh was an important oxidoreductase in these cells; nevertheless, D. ethenogenes cultures could not use formate as an electron donor for reductive dechlorination. Analysis of the gene encoding the large subunit of Fdh indicated that while it was related to other Fdh proteins, its sequence encodes serine rather than cysteine or selenocysteine at a critical position, casting doubt on its function. Overall, genomic and proteomic approaches have provided novel insights into the metabolism of this difficult to culture organism.
Heat shock proteins in the skin
Authors: Jonak C. , Klosner G. , Trautinger F.
Published: July 2006

Synopsis

Heat shock proteins (hsp) are expressed in all cells and organisms. Their expression is induced by heat shock (temperatures above 42°C) and other forms of pathophysiological stress. Elevated levels of hsp protect cells from further stress exposure. Hsp are expressed intracellularly. They are highly conserved throughout evolution indicating hsp being necessary for survival under potentially harmful environmental conditions. Hsp are divided into families according to their molecular weight. The majority of hsp function as molecular chaperones. Chaperone function is characterized by binding to other proteins and mediating their folding, transport and interaction with other molecules. In human epidermis hsp are abundantly expressed and have been linked with functions in cell differentiation and photobiology. Recent research has mainly focused on the 27 and 72 kD hsp that are constitutively expressed in human keratinocytes. ultraviolet radiation (UV)‐induced cell death and sunburn cell formation can be inhibited by previous heat shock exposure and UV itself can induce hsp expression. The expression of the 27 kD hsp (hsp27) in epidermal keratinocytes in situ and in culture correlates with differentiation. Expression of hsp27 increases simultaneously with keratinocyte differentiation. For that reason, hsp27 is described as a marker of epidermal differentiation. Changes in the expression and inducibility of hsp have been linked with ageing. In the skin, recent data indicate that hsp72 expression remains remarkably stable with intrinsic ageing. In contrast, levels of hsp27 have been found to be elevated in sun‐protected aged skin indicating a link between hsp27 expression and age‐dependent epidermal alterations. Regulation of hsp can be modified by pharmacological intervention and the development of safe topical and systemic treatments for the prevention of skin damage and disorders of keratinocyte differentiation can be expected for the future.
Stress and the skin
Authors: Hosoi J.
Published: July 2006

Synopsis

‘The skin is the mirror which reflects the state of the mind.’‘The skin is the window of the mind.’ These have been proverbs since ancient times. It is the topic of this article. Our life became convenient with the information technology these days but too much information often drives us on. We suffer from mental stress rather than physical stress. Since Selye advocated stress reaction, various reactions in the body have been described. Skin is also a target organ of the stress reaction. What the effects of stress are and how stress affects the skin are summarized in this review. Possible use of fragrance for the regulation of the stress reaction is also introduced.
Artocarpus lakoochaheartwood extract as a novel cosmetic ingredient: evaluation of thein vitroanti‐tyrosinase andin vivoskin whitening activities
Authors: Tengamnuay P. , Pengrungruangwong K. , Pheansri I. , Likhitwitayawuid K.
Published: July 2006

Synopsis

The heartwood extract ofArtocarpus lakoochaRoxb. was evaluated for thein vitrotyrosinase inhibitory activity and thein vivomelanin‐reducing efficacy in human volunteers. The IC50of the extract and oxyresveratrol, its major active ingredient, against mushroom tyrosinase was determined to be 0.76 and 0.83 μg mL−1, respectively. The extract dissolved in propylene glycol was subsequently tested in female volunteers using a parallel clinical trial with self‐control (n = 20 per group). The first group received the 0.25% w/vA. lakoochasolution as the test solution, whereas the second and the third group, respectively, received 0.25% licorice extract and 3% kojic acid as the reference solutions in the same solvent. The subjects in each group twice daily applied the test (or reference) solution in one of her upper arm, whereas the remaining arm was treated with only propylene glycol (self‐control) for 12 weeks. The melanin content of each application site was measured using Mexameter every week and calculated as % reduction in melanin content relative to the initial melanin value (% whitening). The value of % whitening was then compared between the product‐treated and the propylene glycol‐treated arms within the same subject using pairedt‐test (α = 0.05). TheA. lakoochaextract was the most effective agent, giving the shortest onset of significant whitening effect after only 4 weeks of application (P < 0.05), followed by 3% kojic acid (6 weeks) and 0.25% licorice extract (10 weeks). The effect also increased with time with maximum whitening observed at week 12 forA. lakoochaextract. When the extract was formulated as an oil‐in‐water emulsion, its whitening efficacy was further enhanced. Daily application of 0.1% w/wA. lakoochalotion to the upper arms (n = 25) and cheeks (n = 15) of volunteers produced significant whitening over the lotion base after 2 and 3 weeks, respectively (P < 0.05). Thus, the preliminary study suggested that the heartwood extract ofA. lakoochamay have a promising potential for use as an effective and economical skin‐whitening agent.
Preventing paediatric obesity; recommendations from a community‐based qualitative investigation
Authors: Tucker P. , Irwin J. D. , Sangster Bouck L. M. , He M. , Pollett G.
Published: July 2006

Childhood obesity is on the rise, and interventions targeted at pre‐school‐aged children are essential for the primary prevention of this disease. Physical activity programming and decreasing screen viewing may be advantageous ways to reduce the early onset of obesity. Parents’ perceptions regarding effective tools and programmes to target obesity are needed to develop an efficacious programme. Ten semi‐structured focus groups were conducted for this qualitative study. Two experienced moderators facilitated all focus groups which were conducted in local day cares, workplaces and play groups, frequented by parents. All focus groups were audio‐recorded and transcribed verbatim. Strategies to ensure trustworthiness of the data were employed. A heterogeneous sample of 71 parents were asked about their pre‐schoolers’ physical activity levels and screen viewing behaviours, in addition to their suggestions for programmes to facilitate the development of healthy habits. Parents agreed that physical activity programming was an ideal way to combat obesity in pre‐schoolers. Programming suggestions included involving parents in the programme planning and operation, increasing accessibility to programmes, providing more facilities, better promotional strategies such as the use of a television commercial, and resources/ideas to engage children in the home. Parents’ programming suggestions lacked discussion around addressing screen viewing as a viable way to attend to the current obesity epidemic. Parents must be made aware of programmes and resources currently available. Modifying current programmes’ scheduling to include morning and afternoon sessions, and arranging for physicians to hand out resources with physical activity ideas, particularly during the winter months, is essential.
Measuring the full economic costs of diet, physical activity and obesity‐related chronic diseases
Authors: Popkin B. M. , Kim S. , Rusev E. R. , Du S. , Zizza C.
Published: July 2006

Most studies that have focused on the costs of obesity have ignored the direct effects of obesity‐related patterns of diet and physical activity. This study reviews the full effects of each component – poor dietary and physical activity patterns and obesity – on morbidity, mortality and productivity. The direct healthcare costs are based on a review of the effects of these factors on key diseases and the related medical care costs of each disease. The indirect costs on reduced disability, mortality and sickness during the period of active labour force participation prior to retirement are also examined. A case study is prepared for China to provide some guidance in the utilization of this review for economic analysis of obesity. The case study shows that the indirect costs are often far more important than the direct medical care costs. The Chinese case study found that the indirect effects of obesity and obesity‐related dietary and physical activity patterns range between 3.58% and 8.73% of gross national product (GNP) in 2000 and 2025 respectively.
Prevalence of obesity in Portugal
Authors: Carmo I. do , Dos Santos O. , Camolas J. , Vieira J. , Carreira M. , Medina L. , Reis L. , Galvão‐Teles A.
Published: July 2006

Obesity is a serious health problem in developed countries. Knowledge of reliable and nationwide representative data is a must for any public health action. The dimension of this problem in Portugal was first assessed in 1995–1998. A similar populational survey using objective anthropometric measures is now being conducted (field work started in January 2003). A total of 6411 subjects aged 18–64 years old have already been measured and their respective body mass indexes (BMIs) calculated. Results from 2003 to 2005 show 38.6% is overweight and 13.8% has obesity. The total of overweight and obesity is 52.4%. In 1995–1998 survey, overweight was 35.2% and obesity was 14.4%. The total was 49.6%. These results suggest an increase of the overweight/obesity (altogether) prevalence in the last 10 years. Actual results show that more than half of the adult sample has excessive weight and 2.4% of the sample has low BMI. Finally, 45.6% of the sample suffers increased health risks because of high waist circumference (≥80 cm for women; ≥94 cm for men). These results highlight the fact that, although obesity was identified as a public health problem one decade ago, action to reduce it does not seem to have been very effective to date.
Sleeping more as a way to lose weight
Authors: Sivak M.
Published: July 2006

Caloric consumption in a society with readily available food is likely to be approximately proportional to the number of hours of being awake. Thus, replacing 1 h of inactive wakefulness (e.g. watching TV), with sleeping is likely to result in a substantial reduction in caloric intake. Calculations are presented to illustrate the possible benefits of such a switch on weight reduction.
Do Friendships and Sibling Relationships Provide Protection against Peer Victimization in a Similar Way?
Authors: Lamarche Véronique , Brendgen Mara , Boivin Michel , Vitaro Frank , Pérusse Daniel , Dionne Ginette
Published: July 2006

Based on the notion that friendship may serve an important protective function against peer victimization, this study examined the moderating effect of reciprocal friends' prosociality on the link between a child's reactive aggression and victimization. The study also investigated whether a similar moderating effect could be found with respect to sibling's prosociality, given that sibling relationships have been found to provide social benefits comparable to friendships. These questions were addressed using a sample of 246 six‐year‐old twin pairs (246 boys and 246 girls). The results showed that a child's own reactive aggression uniquely contributed to the risk of victimization for both boys and girls. The link between reactive aggression and victimization was, however, moderated by reciprocal friends' prosocial behavior and siblings' prosocial behavior, respectively. The results are discussed in terms of their theoretical and prevention‐related implications for children at risk for peer victimization.
Popular and Rejected Children's Reasoning Regarding Negative Emotions in Social Situations: The Role of Gender
Authors: Banerjee Robin , Rieffe Carolien , Terwogt Mark Meerum , Gerlein Ana Maria , Voutsina Maria
Published: July 2006

Two studies compared popular and rejected children's reasoning regarding social interactions involving negative emotions. The first study, with 23 rejected and 23 popular 10‐ to 11‐year‐olds, involved hypothetical social scenarios where a classmate ‘victim’ was likely to experience a negative emotion. Although popular and rejected children both recognized negative emotions and were equally likely to suggest helping behaviour to aid the victim, there were gender effects on the type of helping behaviour suggested. Specifically, popular girls were significantly more likely to offer comforting behaviour than advice whereas popular boys offered advice more than comfort; no such preferences were exhibited by the rejected children. Furthermore, popular girls were significantly more likely than other children to refer to emotional states when justifying their helping response. In the second study, 30 popular and 30 rejected eight‐ to 10‐year‐olds identified the motives behind story characters' efforts to mask negative emotions. Popular girls were more likely to identify the target motives than rejected girls, but no such difference was apparent for the boys. The results are discussed in the light of evidence regarding gender differences in peer interaction patterns.
The Use of Discordant MZ Twins to Generate Hypotheses regarding Non‐shared Environmental Influence on Anxiety in Middle Childhood
Authors: Asbury Kathryn , Dunn Judy , Plomin Robert
Published: July 2006

Non‐shared environmental influence (NSE) has been found to account for around 50 percent of anxiety variance, but specific NSE factors have not been identified. Discordant monozygotic (MZ) twins can be used to generate relevant hypotheses because MZ discordance can be caused by NSE, but not by genes or shared environment. Of 1590 MZ pairs teacher rated for anxiety at the age of seven, we selected the most discordant 19 pairs and conducted interviews with them and their mothers. In explaining discordant anxiety, the mothers emphasised negative school experiences, comparisons within the twin relationship, and illness and accidents. During the maternal interviews neonatal life events, parent–child relationships and peer rejection also emerged as potential NSE sources.
Polyneuropathy with demyelinating features in mixed cryoglobulinemia with hepatitis C virus infection
Authors: Boukhris S. , Magy L. , Senga‐mokono U. , Loustaud‐ratti V. , Vallat J.‐M.
Published: July 2006

Peripheral neuropathy can arise from various mechanisms during hepatitis C virus (HCV) infection, mainly involving associated mixed cryoglobulinemia. The frequency of demyelinating polyneuropathy is probably underestimated in these patients. We report two cases of demyelinating polyneuropathy in HCV‐infected patients. The first case concerned a 76‐year‐old woman followed for hepatitis C associated with a mixed cryoglobulinemia (type II), who developed a chronic progressive distal motor weakness and sensory disturbances concomitant with a raise in serum aspartate aminotransferase (GOT/AST) and alanine aminotransferase (GPT/ALT) levels. Other laboratory studies were normal except for a decrease in the hemolytic fraction of complement to 75 IU (n = 400–520). The second case was a 68‐year‐old woman followed for hepatitis C associated with a mixed cryoglobulinemia (type II), who had sensory disturbances in the lower limbs. Laboratory studies were otherwise unremarkable. Cerebrospinal fluid studies showed a normal protein content without pleocytosis in both patients. In both cases nerve conduction studies were suggestive of a mixed axonal and demyelinating sensorimotor neuropathy. Sural nerve biopsy showed segmental demyelination and severe loss of large myelinated fibers as well as some onion bulb formation in both cases. The two patients subsequently improved, the first with an antiviral treatment and the second with oral steroids.
Examination of the diagnostic validity of ‘headache attributed to whiplash injury’: a controlled, prospective study
Authors: Schrader H. , Stovner L. J. , Obelieniene D. , Surkiene D. , Mickevičiene D. , Bovim G. , Sand T.
Published: July 2006

Acute and chronic headache attributed to whiplash injury are new diagnostic entities in the International Classification of Headache Disorders, second edition. A main objective of the present study was to assess the validity of these nosologic entities by studying the headache pattern in an inception cohort of 210 rear‐end car collision victims and in 210 matched controls. Consecutive drivers involved in rear‐end collisions were identified from the daily records of the Traffic Police Department of Kaunas, Lithuania. A standard self‐report questionnaire was sent to the drivers between 2 and 7 days after the collision, and their passengers were recruited as well. Headache and neck pain were evaluated within 7 days of the collision, at 2 months and 1 year after the collision. A control group of non‐traumatized subjects received questionnaires at the time of the selection and 1 year later. Of the 75 collision victims who developed headache within the first 7 days of the collision, 37 had a clinical picture in accordance with the criteria for acute whiplash headache (i.e., concomitant neck pain) and 38 did not. For acute headache after collision, concomitant neck pain was of no relevance to the headache type or its course. In both these subgroups, migraine and tension‐type headache could be diagnosed in similar proportions and the prognosis after 2 months and 1 year was also similar. Preexisting headache was a strong prognostic factor in both groups for both acute and chronic pain. Compared with the non‐traumatized control group, the 1‐year incidence of new or worsened headache, or of headache improvement, was the same. A likely interpretation of the data is that acute headaches after rear‐end car collisions mainly represent episodes of a primary headache precipitated by the stress of the situation. We conclude that the nosologic validity of both acute and chronic whiplash headache is poor as the headaches, in accordance with the criteria lack distinguishing clinical features and have the same prognosis compared with headaches in a control group.
Stroke complicating pregnancy and the puerperium
Authors: Liang C.‐C. , Chang S.‐D. , Lai S.‐L. , Hsieh C.‐C. , Chueh H.‐Y. , Lee T.‐H.
Published: July 2006

Stroke complicating pregnancy and the puerperium in Taiwan and the comparison between Western and Eastern countries have not been well studied. We identified retrospectively 32 cases of stroke from 66 781 deliveries, including 21 intracranial hemorrhages and 11 cerebral infarctions from 1992 to 2004. The most common causes of intracranial hemorrhage were vascular anomaly (29%), pre‐eclampsia/eclampsia (24%), undetermined (24%) and coagulopathy (19%). The most common causes of cerebral infarction were cardioembolism (36%), cerebral venous thrombosis (27%) and pre‐eclampsia/eclampsia (18%). Perinatal adverse outcome included two stillbirths, nine premature deliveries and four abortions. The compiled results of previous studies and ours revealed that intracranial hemorrhage appeared to be slightly more common in Taiwan (43–69%) than in the Western countries (33–52%). The average maternal mortality rate was 17.8% (range 9–38) with 77.8% due to intracranial hemorrhage. The average incidence of stroke associated with pregnancy and the puerperium was 21.3 per 100 000 deliveries (range 8.9–67.1). Our study, different from the Western countries, showed that intracranial hemorrhage is slightly more common than cerebral infarction. Pre‐eclampsia/eclampsia is an important cause of stroke, however, the possibility of cardioembolism in cerebral infarction and vascular anomaly in intracranial hemorrhage should be studied.
Validation of the deCODE Migraine Questionnaire (DMQ3) for use in genetic studies
Authors: Kirchmann M. , Seven E. , Björnsson Á. , Björnssdóttir G. , Gulcher J. R. , Stefánsson K. , Olesen J.
Published: July 2006

We assessed the reliability of the diagnosis of migraine with aura (MA) and migraine without aura (MO) based on the third edition of the deCODE Migraine Questionnaire (DMQ3) using a physician‐conducted interview as an empirical index of validity. Amongst Danish migraine families recruited from specialist practice we selected 200 cases diagnosed according to the International Classification of Headache Disorders 2nd Edition in a validated physician‐conducted telephone interview: 50 patients with exclusively MA, 50 with both MA and MO, 50 with exclusively MO and 50 controls. A written copy of the DMQ3 was mailed to the participant. The DMQ3‐based diagnosis was compared with the interview‐based diagnosis. Overall, the DMQ3 diagnosed migraine (MA, MO or both) with a sensitivity of 99% (109/110), a specificity of 86% (32/37) and a kappa statistic of 0.89. The most reliable subtype of migraine was MA (with or without co‐occurring attacks of MO) which was diagnosed with a sensitivity of 92% (71/77), a specificity of 93% (65/70) and a kappa statistic of 0.85. Exclusively MO was diagnosed with a sensitivity of 91% (30/33), a specificity of 93% (106/114) and a kappa statistic of 0.80. Weakest was the diagnosis of both MO and MA which was diagnosed with a sensitivity of 63% (24/38), a specificity of 92% (100/109) and a kappa statistic of 0.57. In conclusion, the DMQ3 is a valid tool for diagnosing patients with migraine for genetic studies.
Headache prevalence related to smoking and alcohol use. The Head‐HUNT Study
Authors: Aamodt A. H. , Stovner L. J. , Hagen K. , Bråthen G. , Zwart J.
Published: July 2006

The aim of this study was to examine a possible association between smoking, alcohol and headache in a large population‐based cross‐sectional study. A total of 51 383 subjects completed a headache questionnaire and constituted the ‘Head‐HUNT’ Study. Questionnaire‐based information on smoking was available in 95% and on alcohol in 89% of the individuals. Associations were assessed in multivariate analyses, estimating prevalence odds ratios (ORs) with 95% confidence intervals (CI). Prevalence rates for headache were higher amongst smokers compared with never smokers, most evident for those under 40 years smoking more than 10 cigarettes per day (OR 1.5, 95% CI 1.3–1.6). Passive smoking was also associated with higher headache prevalence. For alcohol use, there was a tendency of decreasing prevalence of migraine with increasing amounts of alcohol consumption compared with alcohol abstinence. Only with regard to symptoms indicating alcohol overuse, a positive association with frequent headache was found. The association between headache and smoking found in the present study raises questions about a causal relationship, e.g. that smoking causes headache or that it allays stress induced by headache. The observed negative association between migraine and alcohol consumption is probably explained by the headache precipitating properties of alcohol.
Serial measurements of plasma homocysteine levels in early and late phases of ischemic stroke
Authors: Haapaniemi E. , Helenius J. , Soinne L. , Syrjälä M. , Kaste M. , Tatlisumak T.
Published: July 2006

High plasma levels of homocysteine (Hcy) may predispose to ischemic stroke (IS), but results of previous studies have been conflicting. We decided to determine in IS patients whether their Hcy levels are elevated, whether levels vary at different time points following stroke, whether levels are associated with stroke severity, outcome, recurrence, etiology, infarct volume, or risk factors, and whether levels are correlated with hemostatic factors or C‐reactive protein values. We measured plasma Hcy levels in 102 consecutive IS patients on admission and at 1 week, 1 month, and 3 months after stroke and once in 102 control subjects. Hemostatic factors were measured in 55 patients. Compared with controls, plasma Hcy levels in patients were significantly lower on admission but not at later time points, with levels increasing by week and remaining at this level for 3 months. Hcy levels showed a positive correlation with age and a negative correlation with Mini‐Mental State Examination (MMSE) scores. Plasma Hcy levels inversely correlated with plasminogen activator inhibitor type‐1. Decreased Hcy levels on admission may reflect the strength of the acute‐phase response rather than a pathogenetic event. The negative correlation between Hcy levels and MMSE scores is more probably age‐related than stroke‐related.
Is uncertain diagnosis a more frequent reason for referring migraine patients to neurologist than other headache syndromes?
Authors: Bekkelund S. I. , Salvesen R.
Published: July 2006

The objective was to assess whether the physician's uncertainty of diagnosis was a more frequent motivation for referring migraine patients than other headache patients to a neurologist. In this population based survey we included 846 consecutive patients referred to and examined at a specialist centre for headache during a period of 2 years. As primary outcome we compared cross‐sectionally frequency of referring patients with migraine and other headaches to neurologist. According to the patients, uncertainty of diagnosis was the main reason for referral to a neurological specialist consultation, i.e. in 51% of cases. Thirty‐nine per cent of patients classified as having migraine were referred because of an uncertain diagnosis, compared with 62% in patient with other headaches (95 CI 0.13–0.33), (P < 0.0001). Headache subtypes other than migraine and younger age were associated to more uncertainty of diagnosis. We conclude that diagnostic uncertainty was an important factor amongst half the headache patients seen by neurologists and uncertain diagnoses as reason for referral was more important amongst patients with non‐migrainous headache.
Tracer Clearance in Radionuclide Cisternography in Patients with Spontaneous Intracranial Hypotension
Authors: Horikoshi T , Ikegawa H , Uchida M , Takahashi T , Watanabe A , Umeda T
Published: July 2006

We semiquantitatively analysed radionuclide cisternography in three patients with spontaneous cerebrospinal fluid (CSF) leakage diagnosed by typical symptoms and magnetic resonance imaging findings before and several months after treatment with epidural blood patch. Radioactivity in the whole CSF space was measured immediately after and at 1, 5, 7 and 24 h after intrathecal injection of 111In-diethylenetriaminepentaacetic acid (DTPA). Initial findings included the vague appearance of leakage in the thoracic spine in two patients, early bladder filling at 1 h in one and a lack of tracer filling into the high cranial convexity in all three. The radioactivity count rapidly decreased within several hours after injection and reached 20± of the initial value at 24 h. In contrast, no rapid decrease was observed after treatment and more than 50± of tracer remained at 24 h after injection. Semiquantitative analysis of tracer clearance may provide additional information in the diagnosis of CSF leakage, especially with no obvious qualitative findings.
Interspecific differences in foraging preferences, breeding performance and demography in herring (Larus argentatus) and lesser black‐backed gulls (Larus fuscus) at a mixed colony
Authors: Kim S.‐Y. , Monaghan P.
Published: July 2006

Herring gulls Larus argentatus and lesser black‐backed gulls Larus fuscus breeding at Walney Island, Cumbria, the largest breeding colony of the two species in the UK, have recently shown very different population trends. The former has declined sharply, whereas numbers of the latter have been maintained for several years. Here we compare aspects of the feeding and breeding ecology of the two species in order to examine whether or not this suggests explanations for their different population trends. Comparison of the ratio of the two species in flight lines leading to different feeding sites and their diet composition showed that the lesser black‐backed gulls fed more at sea and the herring gulls fed more in the intertidal zone. Urban resources were used by both these species. These differences have been consistent over the last three decades. Susceptibility to death from botulism at the breeding colony was the same for the two species. The availability of the intertidal zone for foraging appears to have declined in recent years, and this may have had a more negative impact on the herring gull. However, the breeding success of the two species remains relatively high. This study suggests that differences in foraging behaviour and food availability during the breeding season are unlikely to be responsible for the marked differences in demographic trends in the two species. Changes in local food availability during the winter would be expected to have more effect on the resident herring gull. This work highlights the need for more detailed studies of the ecology of both species during the breeding season and in winter in regions showing differing patterns of population change.
Physicochemical properties of low‐fat and full‐fat Cheddar cheeses
Authors: KÜÇÜKÖNER E , HAQUE Z U
Published: July 2006

Low‐fat (6% fat) and full‐fat (32% fat) Cheddar cheese was manufactured and aged up to 6–9 months at 5°C. The objective was to study the impact of fat on the physicochemical properties of Cheddar cheese. Total soluble nitrogen (TSN) and protein nitrogen (TPSN) in aqueous extracts were determined by the Kjeldahl method. The peptide content of each cheese was determined with reverse phase chromatography (RPC). Low‐fat Cheddar (LFC) had a markedly higher peptide content than full‐fat Cheddar (FFC). The overall peptide quantity increased with age with a marked increase in hydrophobic peptide content. Rheological properties were determined using an Instron Universal Testing Machine. LFC had significantly higher stress values, indicating hard and rubbery texture, than FFC. Furthermore, LFC had lower strain values, indicating crumbliness.
Protein composition of milk from Holstein–Friesian dairy cows and its relationship with the genetic variants A and B of κ‐casein and β‐lactoglobulin (Part I)
Authors: MOLINA L H , KRAMM J , BRITO C , CARRILLO B , PINTO M , FERRANDO A
Published: July 2006

The protein composition and the milk yield (morning milking) were studied in 10 Chilean Holstein–Friesian first lactation dairy cows during the spring season. In addition, the genetic variants A and B of κ‐casein (κ‐CN) and β‐lactoglobulin (β‐Lg) were also identified. The results showed that the casein content is significantly affected by the presence of B variant of κ‐CN. Moreover, positive interactions between the B variant of κ‐CN with AB of β‐Lg and the combination of A κ‐CN with AA of β‐Lg for the total protein content, were found.
Determination of antimutagenic properties of acetone extracted fermented milks and changes in their total fatty acid profiles including conjugated linoleic acids
Authors: GUZEL‐SEYDIM Z B , SEYDIM A C , GREENE A K , TAŞ T
Published: July 2006

Kefir has a unique microbial content that results in complex fermented milk drink. In this study, the antimutagenic power of acetone‐extracted kefir was determined and compared to acetone‐extracted yogurt and milk using different chemical mutagens in the Ames Salmonella microsomal test. Acetone extracts were fat portions of the kefir, yogurt and milk samples. The direct‐acting mutagenic agents methylmethanosulphate (MMS) and sodium azide (SA), and the indirect‐acting mutagenic agents aflatoxin B1 (AFB1), 2‐aminoanthracene (2AA), and 3,2‐dimethyl‐4‐aminobiphenyl (DMAB) were used in Ames test with TA 100 Salmonella typhimurium. Acetone‐extracted fermented milk samples significantly inhibited mutagenicity for different percentages of MMS, SA, AFB1 and 2‐AA. Results indicate that milk also had a lower antimutagenic effect. Additionally, the fatty acid profiles of milk, yogurt and kefir were determined. A two‐step methylation method followed by gas chromatography was used to identify conjugated linoleic acids (CLA). Fermentation slightly affected fatty acid composition. Three isomers of CLA (c9, t11; t10, c12; t9, t11), butyric, palmitic, palmitoleic, oleic acids, which have been proven as antimutagenic components of milk fat, were in higher concentrations in kefir.
Populations, types and biochemical activities of aerobic bacteria and lactic acid bacteria from the air of cheese factories
Authors: SOLDATOU HELENI , PSONI LEFKI , TZANETAKIS NIKOLAOS , LITOPOULOU‐TZANETAKI EVANTHIA
Published: July 2006

This investigation was carried out in order to identify the airborne bacteria in the processing rooms of two creameries that make, mainly, feta cheese and study some of their biochemical activities. Our results suggest that the numbers of airborne micro‐organisms may differ between plants, and the same was observed with the predominant types. The bacteria isolated were primarily Gram‐positive, catalase‐negative cocci, micrococci and bacilli. The predominant lactic microflora were lactococci and mesophilic lactobacilli, commonly found in feta and other traditional Greek cheeses. A significant number of aerobic bacteria could grow at 4°C and in 6.5% NaCl. In addition, both aerobic and lactic acid bacteria (LAB) exhibited acidifying and proteolytic activities. Slow‐ and medium‐acidification ability of LAB strains were found. There was also a tendency of the strains to preferentially degrade αs‐casein. The amounts of amino acids accumulated in the milk were strain dependent. Therefore, there is a possibility that airborne bacteria contaminating cheese milk in the open vats may multiply and contribute to the ripening changes in the cheese.
Selective blockage of Kv1.3 and Kv3.1 channels increases neural progenitor cell proliferation
Authors: Liebau Stefan , Pröpper Christian , Böckers Tobias , Lehmann‐Horn Frank , Storch Alexander , Grissmer Stephan , Wittekindt Oliver H.
Published: July 2006

The modulation of cell proliferation in neural progenitor cells (NPCs) is believed to play a role in neuronal regeneration. Recent studies showed that K+ channel activity influenced cell proliferation. Therefore, we examined NPCs for K+ channels and tested whether NPC self renewing can be modulated by synthetic K+ channel modulators. The whole‐cell K+ current was partly K+ dependent and showed a cumulative inactivating component. Two tetra‐ethyl‐ammonium ion (TEA)‐sensitve K+ currents with different voltage dependencies ( = 65 μm, E50 = −0.3 ± 1.3 mV and  = 8 mm, E50 = −15.2 ± 2.8 mV) and an almost TEA‐insensitive current were identified. Kaliotoxin blocked approximately 50% of the entire K+ currents (IC50 = 0.25 nm). These properties resembled functional characteristics of Kv1.4, Kv1.3 and Kv3.1 channels. Transcripts for these channels, as well as proteins for Kv1.3 and Kv3.1, were identified. Immunocytochemical staining revealed Kv1.3 and Kv3.1 K+ channel expression in almost all NPCs. The blockage of Kv3.1 by low concentrations of TEA, as well as the blockage of Kv1.3 by Psora‐4, increased NPC proliferation. These findings underline the regulatory role of K+ channels on the cell cycle and imply that K+ channel modulators might be used therapeutically to activate endogenous NPCs.
Up‐regulation of P‐glycoprotein expression by glutathione depletion‐induced oxidative stress in rat brain microvessel endothelial cells
Authors: Hong Hao , Lu Ying , Ji Zhao‐Ning , Liu Guo‐Qing
Published: July 2006

Glutathione (GSH) depletion has been implicated in the pathogenesis of neurological diseases. During GSH depletion, cells of the blood–brain barrier (BBB) are subjected to chronic oxidative stress. In this study, we investigated the effect of such stress, produced with the GSH synthesis inhibitorl‐buthionine‐(S,R)‐sulfoximine (BSO), on expression of P‐glycoprotein (Pgp) in primary cultured rat brain microvessel endothelial cells that comprise the blood–brain barrier (BBB). Application of BSO to cell monolayers at concentrations up to 800 µmcaused increases in expression of Pgp. Concentrations ≥ 400 µmBSO decreased cell viability. Application of 200 µmBSO caused a significant increase in Pgp function activity, as assessed by rhodamine 123 (Rh123) accumulation experiments. At this concentration, BSO produced time‐dependent decreases in levels of intracellular GSH and increases in levels of intracellular reactive oxygen species (iROS). The increases were also observed within 48 h following BSO treatment in mdr1a and mdr1b mRNA. Exposure of cells to BSO for 24 h produced maximal effects in the accumulation of iROS, and in expression and function of Pgp. The ROS scavengerN‐acetylcysteine prevented ROS generation and attenuated the changes of both expression and activity of Pgp induced by BSO. Therefore, the transport of Pgp substrates may be affected by changing Pgp expression under conditions of chronic oxidative stress induced by GSH depletion.
Defective processing of neuropeptide precursors in Caenorhabditis elegans lacking proprotein convertase 2 (KPC‐2/EGL‐3): mutant analysis by mass spectrometry
Authors: Husson Steven J. , Clynen Elke , Baggerman Geert , Janssen Tom , Schoofs Liliane
Published: July 2006

Biologically active peptides are synthesized as larger inactive proprotein peptide precursors which are processed by the concerted action of a cascade of enzymes. Among the proprotein convertases, PC2 is widely expressed in neuro‐endocrine tissues and has been proposed to be the major convertase involved in the biosynthesis of neuropeptides. In this study, we have examined the role of the Caenorhabditis elegans orthologue PC2/EGL‐3 in the processing of proprotein peptide precursors. We recently isolated and identified 60 endogenous peptides in the nematode C. elegans by two‐dimensional nanoscale liquid chromatography – quadrupole time‐of‐flight tandem mass spectrometry. In the present study, we compare the peptide profile of different C. elegans strains, including PC2/EGL‐3 mutants. For this purpose, we used an offline approach in which HPLC fractions are analysed by a matrix‐assisted laser desorption ionisation – time of flight mass spectrometer. This differential peptidomic approach unambiguously provides evidence for the role of PC2/EGL‐3 in the processing of FMRFamide‐like peptide (FLP) precursors and neuropeptide‐like protein (NLP) precursors in nematodes.
Oxidative stress‐triggered unfolded protein response is upstream of intrinsic cell death evoked by parkinsonian mimetics
Authors: Holtz William A. , Turetzky Jay M. , Jong Yuh‐Jiin I. , O'Malley Karen L.
Published: July 2006

Oxidative stress is a key player in a variety of neurodegenerative disorders including Parkinson's disease. Widely used as a parkinsonian mimetic, 6‐hydroxydopamine (6‐OHDA) generates reactive oxygen species (ROS) as well as coordinated changes in gene transcription associated with the unfolded protein response (UPR) and apoptosis. Whether 6‐OHDA‐induced UPR activation is dependent on ROS has not yet been determined. The present study used molecular indicators of oxidative stress to place 6‐OHDA‐generated ROS upstream of the appearance of UPR markers such as activating transcription factor 3 (ATF3) and phosphorylated stress‐activated protein kinase (SAPK/JNK) signaling molecules. Antioxidants completely blocked 6‐OHDA‐mediated UPR activation and rescued cells from toxicity. Moreover, cytochrome c release from mitochondria was observed after the appearance of early UPR markers, suggesting that cellular stress pathways are responsible for its release. Mechanistically, the 6‐OHDA‐induced UPR was independent of intracellular calcium changes. Rather, evidence of protein oxidation was observed before the expression of UPR markers, suggesting that the rapid accumulation of damaged proteins triggered cell stress/UPR. Taken together, 6‐OHDA‐mediated cell death in dopaminergic cells proceeds via ROS‐dependent UPR up‐regulation which leads to an interaction with the intrinsic mitochondrial pathway and downstream caspase activation.
GLT‐1 down‐regulation induced by clozapine in rat frontal cortex is associated with synaptophysin up‐regulation
Authors: Bragina Luca , Melone Marcello , Fattorini Giorgia , Torres‐Ramos Monica , Vallejo‐Illarramendi Ainara , Matute Carlos , Conti Fiorenzo
Published: July 2006

In rat frontal cortex, extracellular levels of glutamate are raised by the anti‐psychotic drug clozapine. We have recently shown that a significant reduction in the levels of the glutamate transporter GLT‐1 may be one of the mechanisms responsible for this elevation. Here we studied whether GLT‐1 down‐regulation induced by chronic clozapine treatment is associated with changes in the expression of synaptophysin, synaptosome‐associated protein of 25 kDa (SNAP‐25) and vesicular glutamate transporter 1 (VGLUT1), three major presynaptic proteins involved in neurotransmitter release. Quantitative high‐resolution confocal microscopy studies in vivo showed that GLT‐1 down‐regulation is closely associated with a significant increase in synaptophysin, but not SNAP‐25 and VGLUT1, expression. This was confirmed in vitro studies, and in western blotting studies of synaptophysin, SNAP‐25 and VGLUT1. In addition, our results show that, following clozapine treatment, synaptophysin expression increases in the very cortical regions in which GLT‐1 expression is down‐regulated. These findings suggest that part of the effects of clozapine may be exerted via an action on the presynaptic machinery involved in neurotransmitter release.
Cyclic ADP‐ribose is a second messenger in the lipopolysaccharide‐stimulated activation of murine N9 microglial cell line
Authors: Franco Luisa , Bodrato Nicoletta , Moreschi Iliana , Usai Cesare , Bruzzone Santina , Scarf ì Sonia , Zocchi Elena , De Flora Antonio
Published: July 2006

Lipopolysaccharide, the main component of the cell wall of Gram‐negative bacteria, is known to activate microglial cells following its interaction with the CD14/Toll‐like receptor complex (TLR‐4). The activation pathway triggered by lipopolysaccharide in microglia involves enhanced basal levels of intracellular calcium ([Ca2+]i) and terminates with increased generation of cytokines/chemokines and nitric oxide. Here we demonstrate that in lipopolysaccharide‐stimulated murine N9 microglial cells, cyclic ADP‐ribose, a universal and potent Ca2+ mobiliser generated from NAD+ by ADP‐ribosyl cyclases (ADPRC), behaves as a second messenger in the cell activation pathway. Lipopolysaccharide induced phosphorylation, mediated by multiple protein kinases, of the mammalian ADPRC CD38, which resulted in significantly enhanced ADPRC activity and in a 1.7‐fold increase in the concentration of intracellular cyclic ADP‐ribose. This event was paralleled by doubling of the basal [Ca2+]i levels, which was largely prevented by the cyclic ADP‐ribose antagonists 8‐Br‐cyclic ADP‐ribose and ryanodine (by 75% and 88%, respectively). Both antagonists inhibited, although incompletely, functional events downstream of the lipopolysaccharide‐induced microglia‐activating pathway, i.e. expression of inducible nitric oxide synthase, overproduction and release of nitric oxide and of tumor necrosis factor α. The identification of cyclic ADP‐ribose as a key signal metabolite in the complex cascade of events triggered by lipopolysaccharide and eventually leading to enhanced generation of pro‐inflammatory molecules may suggest a new therapeutic target for treatment of neurodegenerative diseases related to microglia activation.