bluuebunny/crossref_metadata_2025_split huggingface dataset:
The histological analysis, colorimetric evaluation, and chemical quantification of melanin content in ‘suntanned’ fish
Authors: Adachi Kohsuke , Kato Keitaro , Wakamatsu Kazumasa , Ito Shosuke , Ishimaru Katsuya , Hirata Takashi , Murata Osamu , Kumai Hidemi 
Published: September 2005

SummaryHuman melanocytes respond to UV irradiation by increasing the synthesis of melanin. While much is now understood of the pathways governing this process and the nature of the melanin synthesized, little is known of melanins produced by lower vertebrates and their capacity to respond to UV. Here we report that a fish, red seabream, can undergo ‘suntanning’. Histological, colorimetric and chemical assays were performed for suntanned red seabream fish bred in net cages to analyse the melanins and compared with shaded or wild red seabream fish. For color evaluation, the L* values of suntanned fish were dramatically lower than those in the other two groups. Pyrrole‐2,3,5‐tricarboxylic acid (PTCA), an indicator of eumelanin, was detected in suntanned fish at five times higher levels than in shaded or wild fish while 4‐amino‐3‐hydroxyphenyl‐alanine (4‐AHP), a marker for pheomelanin, could not be detected in any of the samples. Histological analysis showed that melanocytes in the suntanned skin enlarged and increased in number to form a monolayer at the surface of the skin. Analysis of L* values and PTCA levels showed quite a high correlation coefficient (r = −0.843). When comparing shaded and wild red seabream fish, the scores were closer but some significant differences were still found in some body areas. These results indicate that eumelanin accumulates in suntanned fish during the increase in skin color, which is induced by sunlight, presumably by ultraviolet radiation.
Expression of human β‐defensin‐3 in gingival epithelia
Authors: Lu Qian , Samaranayake Lakshman P. , Darveau Richard P. , Jin Lijian 
Published: September 2005

Objective:  This study aimed to investigate the expression patterns of the newly discovered human β‐defensin‐3 (hBD‐3) in human gingiva.Background:  Human β‐defensins (hBDs) are a group of small, broad‐spectrum, cationic antimicrobial peptides. Our recent study showed that the expression levels of hBD‐1 and 2 peptides were associated with periodontal conditions.Methods:  A total of 49 gingival biopsies were collected, including 33 samples from 21 patients with chronic periodontitis and 16 samples from 16 periodontally healthy subjects. The expression of hBD‐3 was detected by immunohistochemistry and in situ hybridization. Double staining was undertaken to identify hBD‐3 peptide‐positive cells, using CD‐1a and cytokeratin 20 as markers for Langerhans cells and Merkel cells, respectively.Results:  hBD‐3 peptide was detected in 88% of the samples, which was confined to the gingival epithelia. In healthy control subjects, hBD‐3 peptide was more frequently detected in the basal layer as compared to the patients (53% vs. 18%, p < 0.05). In patients, hBD‐3 expression extended from the basal layer to the spinous layers (82%), in which hBD‐3 was confined to the basal and deep spinous layers in clinically healthy tissues from patients, whereas it extended to the superficial spinous layers in pocket tissues from patients (0% vs. 50%, p < 0.05). In both groups, hBD‐3 peptide was expressed not only in gingival keratinocytes, but also in Langerhans cells and Merkel cells. hBD‐3 transcripts were detected in 90% of the samples and they were confined to the basal and/or suprabasal layers of gingival epithelia.Conclusions:  This study shows that hBD‐3 is frequently expressed in gingival epithelia. The appropriate expression of hBD‐3 peptide may contribute to the maintenance of periodontal homeostasis, possibly through its antimicrobial effect and promotion of adaptive immune responses.
Prevention of fatal pulmonary embolism and mortality in surgical patients
Authors: Wolf Helmut , Kakkar Ajay K. , Fareed Jawed , Encke Albrecht , Haas Sylvia 
Published: September 2005

SummaryThe incidences of fatal pulmonary embolism and death in surgical patients receiving low-molecular-weight heparin thromboprophylaxis have not been previously determined in large, adequately designed clinical trials and information on the relative efficacy and safety of unfractionated and low-molecular-weight heparin in preventing these clinical endpoints is not available. In a double-blind study, 23078 surgical patients randomly received the low-molecular-weight heparin, certoparin (3000 anti Xa IU) subcutaneously once-daily, or unfractionated heparin (5000 IU) subcutaneously three-times daily, for a minimum of 5 days. The primary outcome measure, autopsy-proven fatal pulmonary embolism recorded up to 14 days after the end of prophylaxis, occurred in 0.152% (95% confidence interval (CI) 0.10, 0.20%; 35of 23078 patients) of cases, with no significant difference between the certoparin-treated patients (0.147% (95% CI 0.077, 0.217%; 17 of 11542 patients) and patients treated with unfractionated heparin (0.156% (95% CI 0.084, 0.228%; 18 of 11,536 patients, P=0.868). The autopsy rate was 70.2%. Comparing mortality, there was no significant difference between the groups (1.44% [166 of 11542 certoparin patients] versus 1.27% [146 of 11536 unfractionated heparin patients]; P=0.279). The safety profiles of both treatment groups were similar. Once-daily certoparin and three-times daily unfractionated heparin are equally effective and safe in reducing fatal pulmonary embolism and death to low levels in surgical patients and mirror the findings of comparative efficacy studies using surrogate endpoints.
Evaluation of perioperative fatal pulmonary embolism and death in cancer surgical patients
Authors: Haas Sylvia , Wolf Helmut , Encke Albrecht , Kakkar Ajay K. 
Published: September 2005

SummaryCancer patients undergoing surgery are at a high risk of venous thromboembolism, but few studies have described the rate of autopsy-confirmed fatal pulmonary embolism after heparin thromboprophylaxis. In a post hoc analysis of a randomized study (MC-4), which compared the efficacy and safety of certoparin (3000 anti-Xa IU, subcutaneously, once-daily) with unfractionated heparin (5000 IU, subcutaneously, three-times daily) in 23078 patients undergoing surgery lasting more than 30 min, the incidence of autopsy-confirmed fatal pulmonary embolism, death and bleeding in the cancer patients (n=6124) was compared with non-cancer patients (n=16954). Fatal pulmonary embolism was significantly more frequent in cancer patients (0.33% [20/6124]) than in non-cancer patients (0.09% [15/16954], relative risk (RR), 3.7 [95% confidence intervals (CI), 1.80, 7.77], p=0.0001) at 14 days post-prophylaxis. Perioperative mortality was also significantly higher in cancer patients than in noncancer patients (3.14% [192/6124] vs. 0.71% [120/16954], RR, 4.54 [95% CI, 3.59, 5.76], p=0.0001), as were blood loss (p<0.0001), and transfusion requirements (p<0.0001). Prevention of venous thromboembolism in cancer surgical patients remains a clinical challenge.
High dose dexamethasone increases circulating P-selectin and von Willebrand factor levels in healthy men
Authors: Cvitko Tuende , Winter-Fabry Astrid , Petroczi Karin , Quehenberger Peter , Blann Andrew D. , Jilma Bernd 
Published: September 2005

SummaryAlthough glucocorticoids are widely used in a number of inflammatory disorders associated with endothelial and platelet activation, their effect on the endothelium and platelets in humans remain poorly defined. Hence, we measured changes of a specific endothelial cell marker (von Willebrand factor [vWF]) and of a platelet marker (soluble P-selectin) by infusing therapeutic doses of dexamethasone (0.04 mg/kg and 1.0 mg/kg b.i.d on two days) or placebo into nine healthy men. Venous citrated plasma was obtained before infusion, and at 24 and 48 h. Compared to baseline levels, we found increased levels of vWF at both time points at the higher dose (p=0.011). Plasma levels of sP-selectin rose at 48 h after the high dose (p=0.017). Human umbilical endothelial cells were cultured in the presence or absence of de-xamethasone (0, 0.01, 1 μM), to determine the possible mechanism for the increase in vWF. The vWF-mRNA levels as quantified by RT-PCR increased 2-fold (p<0.05), and the vWF-concentrations in cell lysates increased by 38% (p<0.05), whereas the vWF-concentrations in the supernatants were unaffected. In summary, high dose DEXA increases sP-selectin and vWF. The probable underlying mechanism for the latter was a DEXA induced up-regulation of vWF-mRNA transcription. Together, this indicates that high dose glucocorticoids may enhance haemostasis, which could be beneficial under certain conditions, but which may also contribute to adverse vascular events by increasing platelet activation and vWF dependent thrombosis.
Microparticles from apoptotic vascular smooth muscle cells induce endothelial dysfunction, a phenomenon prevented by β3-integrin antagonists
Authors: Essayagh Sanah , Brisset Anne-Cecile , Terrisse Anne-Dominique , Dupouy Dominique , Tellier Lise , Navarro Chantal , Arnal Jean-François , Siè Piérre 
Published: September 2005

SummaryFragile atherosclerotic plaques are rich in apoptotic smooth muscle cells (SMCs) and macrophages, generating microparticules (MPs) which accumulate locally and may be released in blood in case of mechanical or spontaneous plaque disruption. Besides being highly procoagulant, this material may interact with downstream endothelium. Using a model of mouse aorta vaso-reactivity, we have investigated the effects of apoptotic MPs preparedin vitro from Fas-ligand sensitive SMCs. Short-term preincubation of aorta rings with the MPs dose-dependently reduced the vasodilatory response to acetylcholine dependent on the endothelium. This effect was prevented by the addition of abxicimab or eptifibatide, indicating a role for a β3 integrin in thisprocess. We further investigated its mechanism using cultured endothelial cells. The MPs were found to bind to the cells and to inhibit the production and the release of nitric oxide (NO) in response to bradykinin. This phenomenom was redox sensitive, independent of the generation of activated coagulation proteases, and was abrogated when the MPs were pretreated by trypsin. The metabolic effects of MPs were prevented by the addition of eptifibatide. Taken together, these results suggest a potential, platelet-independent, mechanism for the improvement of microvascular perfusion observed with β3-integrin antagonists.
Circulating endothelial cells in acute ischaemic stroke
Authors: Nadar Sunil K. , Lip Gregory Y. H. , Lee Kaeng W. , Blann Andrew D. 
Published: September 2005

SummaryIncreased numbers of CD146-bearing circulating endothelial cells (CECs) in the peripheral blood probably represent the most direct evidence of endothelial cell damage. As acute ischaemic strokes are associated with endothelial abnormalities, we hypothesised that these CECs are raised in acute stroke, and that they would correlate with the other indices of endothelial perturbation, i.e. plasma von Willebrand factor (vWf) and soluble E-selectin. We studied 29 hypertensive patients (19 male; mean age 63 years) who presented with an acute stroke and compared them with 30 high risk hypertensive patients (21 male; mean age 62 years) and 30 normotensive controls (16 male; mean age 58 years). CECs were estimated by CD146 immunobead capture, vWf and soluble E-selectin by ELISA. Patients with an acute ischaemic stroke had significantly higher numbers of CECs/ml of blood (p<0.001) plasma vWf (p=0.008) soluble E-selectin (p=0.002) and higher systolic blood pressure (SBP) as compared to the other groups. The number of CECs significantly correlated with soluble E-selectin (r=0.432, p<0.001) and vWf (r=0.349, p=0.001) but not with SBP (r=0.198, p=0.069). However, in multivariate analysis, only disease group (i.e. health, hypertension or stroke) was associated with increased CECs. Acute ischaemic stroke is associated with increased numbers of CECs. The latter correlate well with established plasma markers of endothelial dysfunction or damage, thus unequivocally confirming severe vasculopathy in this condition. However, the greatest influence on CECs numbers was clinical group.
Agonists of toll-like receptor (TLR)2 and TLR4 are unable to modulate platelet activation by adenosine diphosphate and platelet activating factor
Authors: Ward Jon R. , Bingle Lynne , Judge Heather M. , Brown Simon B. , Storey Robert F. , Whyte Moira K. B. , Dower Steven K. , Buttle David J. , Sabroe Ian 
Published: September 2005

SummaryInappropriate platelet activation is a feature of acute and chronic diseases such as disseminated intravascular coagulation (DIC) and atherosclerosis. Since proinflammatory microbial-derived agonists can be involved in the pathogenesis of these diseases, we examined the potential role ofTLR4 (mediating responses to LPS) andTLR2 (which responds to bacterial lipopeptides) in platelet activation. Our data suggested low-level expression of TLR2 andTLR4 on platelets, determined by flow cytometry, and we also observed expression of TLR4 on a megakaryocytic cell line by both flow cytometry and immunohistochemistry. Stimulation of the platelets with the TLR4 agonist LPS, and the synthetic TLR2 agonist Pam3CSK4, resulted in no platelet aggregation, no increase in CD62P surface expression and no increase in the cytosolic concentration of Ca2+. The TLR agonists were also unable to directly activate platelets primed with epinephrine, or pretreated with a low concentration ofADP or PAF. Pretreatment of platelets with LPS or Pam3CSK4 also failed to modulate the platelet response to submaximal concentrations of the classical platelet agonists ADP and PAF. We conclude that theTLR agonists LPS and Pam3CSK4 have no direct effect on platelet activation and that platelet TLRs may be a remnant from megakaryocytes. TLR2 and TLR4 agonists are thought to have a significant role in diseases such as atherosclerosis and DIC, but our research suggests that this is through a mechanism other than direct platelet activation or by modification of platelet responses to other agonists.
Indication of a role of plasminogen activator inhibitor type 1 in protecting murine fibrosarcoma cells against apoptosis
Authors: Rømer Maria Unni , Due Anne Kirkebjerg , Larsen Jørgen Knud , Hofland Kenneth Francis , Christensen Ib Jarle , Buhl-Jensen Peter , Almholt Kasper , Nielsen Ole Lerberg , Lademann Ulrik , Brünner Nils 
Published: September 2005

SummaryIn a number of cancer types high tumor tissue levels of plasminogen activator inhibitor type 1 (PAI-1) protein are strongly associated with shorter cancer patient survival. This association has been intriguing since PAI-1 is known to inhibit urokinase plasminogen activator (uPA) that converts plasminogen to plasmin, which is actively involved in tumor progression and invasion. In order to further explore the biological role of PAI-1 in cancer, we have prepared fibroblasts from PAI-1 gene deficient mice and from their wild type littermates. From these fibroblasts fibrosarcoma cell lines were established and characterized. Both types of fibroblasts underwent spontaneous transformation as indicated by aneuploidy, immortalization, clonogenicity in soft agar and tumor formationin vivo. While both PAI-1 deficient and PAI-1 expressing cell lines showed similar proliferation ratesin vitro, cells devoid of PAI-1 were significantly more sensitive to apoptotic stimuli. When inoculated subcutaneously into nude mice PAI-1 expressing cells rapidly established tumors, while PAI-1 deficient cells had a significantly longer lag-phase before they started to grow (p<0.0001). The present study suggests that PAI-1,besides its uPA inhibiting function, has a role in cancer progression by protecting tumor cells from undergoing apoptosis.
Nitric oxide differentially regulates proliferation and mobilization of endothelial progenitor cells but not of hematopoietic stem cells
Authors: Özüyaman Burcin , Ebner Petra , Niesler Ulrike , Ziemann Jutta , Kleinbongard Petra , Jax Thomas , Gödecke Axel , Kelm Malte , Kalka Christoph 
Published: September 2005

SummaryTo investigate the role of nitric oxide in controlling endothelial progenitor (EPC) and hematopoietic stem cell (HSC) mobilization, wild-type mice, L-NAME treated WT and eNOS-/- mice received either PBS or G-CSF for 5 days. Under unstimulated conditions bone marrow of either L-NAME treated WT and eNOS-/- mice, representing acute and chronic NO-deficiency, showed higher CD34+Flk-1+ EPC numbers compared to their WT littermates. Furthermore, CD34+Flk-1+ progenitors under NO-deficient conditions showed a higher cell turn over since the proliferation and apoptosis activity underin vivo as well as in vitro conditions were enhanced. In line with this finding bone marrow derived EPC differentiation towards endothelial cells was modulated in an NO-dependent manner. Administration of G-CSF resulted in an increase of EPC within the bone marrow of WT animals with a consecutive release of these cells into the peripheral circulation. Under NO-deficient conditions G-CSF failed to increase EPC numbers. In contrast, the HSC population c-kit+Lin− was not influenced by nitric oxide. Thus, NO differentially supports the mobilization of the endothelial committed progenitor subpopulation in bone marrow but does not have an effect on HSCin vivo.
Site-directed mutagenesis of coumarin-type anticoagulant-sensitive VKORC1
Authors: Rost Simone , Fregin Andreas , Hünerberg Mirja , Bevans Carville G. , Müller Clemens R. , Oldenburg Johannes 
Published: September 2005

SummaryCoumarin and homologous compounds are the most widely used anticoagulant drugs worldwide. They function as antagonists of vitamin K, an essential cofactor for the posttranslational γ-glutamyl carboxylation of the so-called vitamin K-dependent proteins. As vitamin K hydroquinone is converted to vitamin K epoxide (VKO) in every carboxylation step, the epoxide has to be recycled to the reduced form by the vitamin K epoxide reductase complex (VKOR). Recently, a single coumarin-sensitive protein of the putativeVKOR enzyme complex was identified in humans (vitamin K epoxide reductase complex subunit 1, VKORC1). Mutations inVKORC1 result in two different phenotypes: warfarin resistance (WR) and multiple coagulation factor deficiency type 2 (VKCFD2). Here, we report on the expression of site-directed VKORC1 mutants, addressing possible structural and functional roles of all seven cysteine residues (Cys16, Cys43, Cys51, Cys85, Cys96, Cys132, Cys135), the highly conserved residue Ser/Thr57, and Arg98, known to cause VKCFD2 in humans. Our results support the hypothesis that the C132-X-X-C135 motif inVKORC1 comprises part of the redox active site that catalyzes VKO reduction and also suggest a crucial role for the hydrophobicThr-Tyr-Ala motif in coumarin binding. Furthermore, our results support the concept that different structural components of VKORC1 define the binding sites for vitamin K epoxide and coumarin.
Circulating endothelial cells in atrial fibrillation with and without acute cardiovascular disease
Authors: Freestone Bethan , Chong Aun Yeong , Nadar Sunil , Lee Kaeng W. , Blann Andrew D. , Lip Gregory Y. H. 
Published: September 2005

SummaryNormal adults have very few circulating endothelial cells (CECs) in their blood, but increased levels have been shown in association with conditions associated with endothelial damage such as myocardial infarction and stroke. As atrial fibrillation (AF) is associated with a hypercoagulable state and abnormalities of plasma indices of endothelial damage/dysfunction, we hypothesised that CECs would also be raised in this condition, and would correlate with these plasma markers. We measured CECs (by immunofluoresence) as an indicator of frank endothelial damage, alongside 3 plasma indices of endothelial perturbation: von Willebrand factor (vWf), soluble E-selectin and soluble thrombomodulin (sTM) (all ELISA) in 28 patients with chronic ‘stable’ AF, 63 patients with AF plus an acute cardiovascular or cerebrovascular event as positive controls, and 20 healthy subjects in sinus rhythm as negative controls. Chronic ‘stable’AF patients had significantly higher levels of plasma vWf (p<0.001),but comparable numbers of CECs (p=0.1638) in comparison to healthy controls. In patients with AF associated with an acute cardiovascular or cerebrovascular event, levels of CECs (p<0.0001) and sTM (p=0.004), but not vWf or sEsel, were significantly increased in comparison to chronic‘stable’AF patients. Patients with uncomplicated AF have abnormal systemic endothelial damage/dysfunction, as evident by increased plasma vWf levels, but normal numbers of CECs, compared to subjects in sinus rhythm. However, following clinical complications, such as stroke or significant haemodynamic compromise, further endothelial disturbance (as indicated by high levels of sTM and CECs) suggests additional endothelial damage.
D-dimer levels in combination with residual venous obstruction and the risk of recurrence after anticoagulation withdrawal for a first idiopathic deep vein thrombosis
Authors: Legnani Cristina , Cini Michela , Guazzaloca Giuliana , Palareti Gualtiero , Cosmi Benilde 
Published: September 2005

SummaryWe assessed the predictive value of D-dimer levels in combination with residual venous obstruction (RVO) for recurrent venous thromboembolism (VTE) in a prospective cohort of outpatients after oral anticoagulant therapy (OAT) suspension for a first episode of idiopathic proximal deep vein thrombosis of the lower limbs during a 2-year follow-up. Patients (n=400) were enrolled on the day of OAT suspension when RVO was determined by compression ultrasonography (present in 48.6% of patients). D-dimer (cut-off value: 500 ng/mL) was measured 30±10 days afterwards (abnormal in 56.4% of patients). The overall recurrence rate was 16.7% (67/400; 95% confidence intervals - CI -:13–21%). The multivariate hazard ratio (HR) for recurrence was 3.32 (95% CI:1.78–6.75; p > 0.0001) for abnormal D-dimer compared to normal D-dimer and 1.2 (95% CI:0.72–2.07; p>0.05) for RVO compared to absent RVO. The recurrence rate was 5.7% (95% CI:2–13%) and 10.4% (95% CI:6–18%), respectively, for normal D-dimer either without or with RVO, 22.9% (95% CI:14–33%) and 25.9% (95% CI: 18–35%), respectively, for abnormal D-dimer, either without or with RVO. When compared with normal D-dimer without RVO, the multivariate HR for recurrence was similar for abnormal D-dimer either with RVO (4.76 – 95% CI:1.78–12.8) or without RVO (4.3–95%:1.56–11.88). Abnormal D-dimer at one month after OAT withdrawal is an independent risk factor for recurrent VTE, while RVO at the time of OAT withdrawal, either with normal or abnormal D-dimer after one month, does not influence the risk of recurrence.