bluuebunny/crossref_metadata_2025 huggingface dataset:
Bifidobacterium longum PL03, Lactobacillus rhamnosus KL53A, and Lactobacillus plantarum PL02 in the Prevention of Antibiotic-Associated Diarrhea in Children: A Randomized Controlled Pilot Trial
Authors: Szymański Henryk , Armańska Małgorzata , Kowalska-Duplaga Kinga , Szajewska Hania
Published: August 2008

Aim: To determine the efficacy of a combination of Bifidobacterium longum PL03, Lactobacillus rhamnosus KL53A and Lactobacillus plantarum PL02 for the prevention of antibiotic-associated diarrhea in children. Methods: Seventy-eight children (age: 5 months to 16 years) with otitis media, and/or respiratory tract infections, and/or urinary tract infections were enrolled in a double-blind randomized control trial in which they received standard antibiotic treatment plus a food supplement containing 108 colony-forming units of B. longum, L. rhamnosus and L. plantarum (n = 40) or a placebo (n = 38) orally twice daily for the duration of antibiotic treatment. Results: Patients receiving probiotics had a similar rate of diarrhea (≧3 loose or watery stools/day for ≧48 h occurring during or up to 2 weeks after the antibiotic therapy) as those receiving placebo (relative risk 0.5, 95% CI 0.06–3.5). The mean number of stools per day was significantly lower in the experimental group (mean difference –0.3 stool/day, 95% CI –0.5 to –0.07). No adverse events were reported. Conclusion: The administration of the 3 probiotics did not significantly alter the rate of diarrhea, although it reduced the frequency of stools per day. As the overall frequency of diarrhea was surprisingly low, these results should be interpreted with caution.
Health-Related Quality of Life of Adults with Childhood Diagnosis of Specific Language Impairment
Authors: Arkkila E. , Räsänen P. , Roine R.P. , Sintonen H. , Vilkman E.
Published: August 2008

Objective: The health-related quality of life (HRQoL) of adults with childhood diagnosis of specific language impairment (SLI) was evaluated. Patients and Methods: Fifty-two patients with a childhood diagnosis of SLI were asked to fill out the 15D-HRQoL questionnaire. The patients were selected from a hospital setting from psychological examination reports showing a below-average verbal intelligence quotient (VIQ) and a normal performance intelligence quotient (PIQ). The 15D was completed adequately by 33 patients (response rate 63.5%). The results were compared with those of age- and gender-matched controls. Results: The total 15D score of the study group was lower than that of population controls, but the difference was not statistically significant. The study group performed significantly worse on the dimensions of speech, usual activities, mental function and distress. Gender or PIQ in childhood was not correlated with the 15D scores or dimension scores in adulthood. Childhood VIQ was associated with the dimensions of mental function (p < 0.01) and usual activities (p < 0.05). Conclusion: Our results support earlier findings that SLI affects the entire life span. Identification of prognostic factors and a prolonged follow-up of SLI patients could improve the QoL of these patients.
Magnesium Ions and Opioid Agonist Activity in Streptozotocin-Induced Hyperalgesia
Authors: Bujalska Magdalena , Malinowska Ewelina , Makulska-Nowak Helena , Gumułka Stanisław Witold
Published: August 2008

Streptozotocin-induced hyperglycemia accompanied by a chronic decrease in the nociceptive threshold is considered a useful model of experimental hyperalgesia. We examined (1) the effect of the opioid receptor agonists and (2) the effect of the magnesium ions (Mg2+) on the antinociceptive action of opioid agonists in a diabetic neuropathic pain model. When administered alone, opioid agonists like morphine (5 mg/kg i.p.) and fentanyl (0.0625 mg/kg i.p.), as well as the partial agonist buprenorphine (0.075 mg/kg) had only little effect on streptozotocin-induced hyperalgesia. However, pretreatment with Mg2+ at a dose of 40 mg magnesium sulfate/kg i.p. markedly enhanced the analgesic activity of all three investigated opioids. Practical aspects of co-administration of magnesium and opioids in diabetic neuropathy are discussed.
Nine Months to Progression Using Fourth-Line Liposomally Encapsulated Paclitaxel against Hepatocellular Carcinoma
Authors: Christopeit Maximilian , Lenz Georg , Forstpointner Roswitha , Bartelheim Kerstin , Kühnbach Robert , Naujoks Kurt , Schalhorn Andreas
Published: August 2008

Background: Hepatocellular carcinoma (HCC) is the third most common cause of cancer deaths. Difficulties to diagnose HCC at early stages remain the major obstacle to curative (surgical) therapy. Therapy in advanced stages has to be considered palliative. In this situation, a considerable amount of attention should be paid to innovative treatment strategies, e.g. including antiangiogenetic drugs. Results: We report on the successful treatment of a patient suffering from progressive HCC with a novel drug (EndoTAG®-1, formerly named LipoPac®) currently investigated in phase II studies. This drug consists of liposomally encapsulated paclitaxel. Its liposomal formulation favors the drug’s adherence to the tumor neovasculature, in effect starving the tumor. Conclusions: EndoTAG®-1 stopped tumor progression for 9 months in our patient. This, along with successes observed testing this drug against other indications, makes it a suitable candidate for future clinical trials.
Distinct Compartmentalization of Dentin Matrix Protein 1 Fragments in Mineralized Tissues and Cells
Authors: Maciejewska Izabela , Qin Disheng , Huang Bingzhen , Sun Yao , Mues Gabrielle , Svoboda Kathy , Bonewald Lynda , Butler William T. , Feng Jerry Q. , Qin Chunlin
Published: August 2008

Dentin matrix protein 1 (DMP1) has been shown to be critical for the formation of dentin and bone. However, the precise pathway by which DMP1 participates in dentinogenesis and osteogenesis remains to be clarified. DMP1 is present in the extracellular matrix of dentin and bone as processed NH2- and COOH-terminal fragments. The NH2-terminal fragment occurs as a proteoglycan, whereas the COOH-terminal fragment is highly phosphorylated. The differences in biochemical properties suggest that these fragments may have different tissue and cell distribution in association with distinct functions. In this study, we analyzed the distribution of the NH2- and COOH-terminal fragments of DMP1 in tooth, bone, osteocytes as well as MC3T3-E1 and HEK-293 cells. Immunohistochemical analyses were performed using antibodies specific to the NH2- or COOH-terminal region of DMP1. Clear differences in the distribution of these fragments were observed. In the teeth and bone, the NH2-terminal fragment was primarily located in the nonmineralized predentin and cartilage of the growth plate, while the COOH-terminal fragment accumulated in the mineralized zones. In osteocytes, the NH2-terminal fragment appeared more abundant along cell membrane and processes of osteocytes, while the COOH-terminal fragment was often found in the nuclei. This pattern of distribution in cellular compartments was further confirmed by analyses on MC3T3-E1 and HEK-293 cells transfected with a construct containing DMP1 cDNA. In these cell lines, the COOH-terminal fragment accumulated in cell nuclei, while the NH2-terminal fragment was in the cytosol. The different distribution of DMP1 fragments indicates that these DMP1 variants must perform distinct functions.
Blocking of Proteolytic Processing and Deletion of Glycosaminoglycan Side Chain of Mouse DMP1 by Substituting Critical Amino Acid Residues
Authors: Peng Tao , Huang Bingzhen , Sun Yao , Lu Yongbo , Bonewald Lynda , Chen Shuo , Butler William T. , Feng Jerry Q. , D’Souza Rena N. , Qin Chunlin
Published: August 2008

Dentin matrix protein 1 (DMP1) is present in the extracellular matrix (ECM) of dentin and bone as processed NH2- and COOH-terminal fragments, resulting from proteolytic cleavage at the NH2 termini of 4 aspartic acid residues during rat DMP1 processing. One cleavage site residue, Asp181 (corresponding to Asp197 of mouse DMP1), and its flanking region are highly conserved across species. We speculate that cleavage at the NH2 terminus of Asp197 of mouse DMP1 represents an initial, first-step scission in the whole cascade of proteolytic processing. To test if Asp197 is critical for initiating the proteolytic processing of mouse DMP1, we substituted Asp197 with Ala197 by mutating the corresponding nucleotides of mouse cDNA that encode this amino acid residue. This mutant DMP1 cDNA was cloned into a pcDNA3.1 vector. Data from transfection experiments indicated that this single substitution blocked the proteolytic processing of mouse DMP1 in HEK-293 cells, indicating that cleavage at the NH2 terminus of Asp197 is essential for exposing other cleavage sites for the conversion of DMP1 to its fragments. The NH2-terminal fragment of DMP1 occurs as a proteoglycan form (DMP1-PG) that contains a glycosaminoglycan (GAG) chain. Previously, we showed that a GAG chain is linked to Ser74 in rat DMP1 (Ser89 in mouse DMP1). To confirm that mouse DMP1-PG possesses a single GAG chain attached to Ser89, we substituted Ser89 by Gly89. Data from transfection analysis indicated that this substitution completely prevented formation of the GAG-containing form, confirming that DMP1-PG contains a single GAG chain attached to Ser89 in mouse DMP1.
Avian Eggshell Structure and Osteopontin
Authors: Chien Y.-C. , Hincke M.T. , McKee M.D.
Published: August 2008

The avian eggshell primarily consists of calcium carbonate mineral (calcite) and matrix proteins. Here we review matrix-mineral relationships in the eggshell at the ultrastructural level using scanning and transmission electron microscopy, and describe the distribution of osteopontin (OPN) as determined by colloidal gold immunolabeling for OPN. A rich protein network integrated within the calcitic structure of the eggshell shows variable, region-specific organization that included layered fibrous planar sheets of matrix, thin filamentous threads, thin film-like surface coatings, vesicular structures and isolated proteins residing on cleaved {104} crystallographic faces of the eggshell calcite. Except for the vesicular structures, these matrix structures all immunolabeled strongly for OPN. Given the potent mineralization- inhibiting function of OPN, we discuss how this protein might regulate eggshell growth rate and inhibit calcification at specific compartmental boundaries to provide eggshell form.
Identification of Peptides with Targeted Adhesion to Bone-Like Mineral via Phage Display and Computational Modeling
Authors: Segvich Sharon , Biswas Subhashis , Becker Udo , Kohn David H.
Published: August 2008

The challenges in engineering bone scaffolds reflect the complexity of bone as an organ. The organic-inorganic hybrid system design aims to provide signals within a conductive apatite layer to promote cell adhesion, proliferation and ultimately differentiation into bone tissue. Dual functioning peptides designed to specifically adhere to the apatite layer, while promoting cell adhesion via cell recognition sequences, may increase cell adhesion, leading to increased osteogenesis. The aim of this study is to identify peptide sequences with preferential adsorption towards apatite-based materials. Bone-like mineral films and hydroxyapatite disks were panned with a phage library to elucidate peptide sequences with favorable adsorption. Peptide sequences were analyzed using the web-based biotechnology tool RELIC and validated with a modified ELISA, in addition to being investigated using a newly developed method of high-throughput computational modeling. Peptides having the highest affinity and greatest potential to be incorporated into a dual functioning peptide design are APWHLSSQYSRT, VTKHLNQISQSY and STLPIPHEFSRE. These experiments provide a method of rationally designing peptides that adhere to apatite and that may improve bone tissue regeneration. This work also provides structure for investigating peptide/protein adsorption on apatite substrates with varied carbonate, or other impurity, content.
Dentin Noncollagenous Matrix Proteins in Familial Hypophosphatemic Rickets
Authors: Gaucher Céline , Boukpessi Tchilalo , Septier Dominique , Jehan Frédéric , Rowe Peter S. , Garabédian Michèle , Goldberg Michel , Chaussain-Miller Catherine
Published: August 2008

Familial hypophosphatemic rickets is transmitted in most cases as an X-linked dominant trait and results from the mutation of the PHEX gene predominantly expressed in osteoblast and odontoblast. Patients with rickets have been reported to display important dentin defects. Our purpose was to explore the structure, composition and distribution of noncollagenous proteins (NCPs) of hypophosphatemic dentin. We collected teeth from 10 hypophosphatemic patients whose mineralization occurred either in a hypophosphatemic environment or in a corrected phosphate and vitamin environment. Teeth were examined by scanning electron microscopy, immunohistochemistry and Western blot analysis. An abnormal distribution (accumulation in interglobular spaces) and cleavage of the NCPs and particularly of matrix extracellular phosphoglycoprotein were observed in deciduous dentin. In contrast, it was close to normal in permanent dentin mineralized under corrected conditions. In conclusion, dentin mineralization in a corrected phosphate and vitamin D environment compensates the adverse effect of PHEX mutation.
Immunogold Labeling of Amelogenin in Developing Porcine Enamel Revealed by Field Emission Scanning Electron Microscopy
Authors: Du Chang , Fan Daming , Sun Zhi , Fan Yuwei , Lakshminarayanan Rajamani , Moradian-Oldak Janet
Published: August 2008

The present study describes a method using immunohistochemical labeling in combination with high-resolution imaging (field emission scanning electron microscopy) to investigate the spatial localization of amelogenins on apatite crystallites in developing porcine enamel. Cross-sections of developing enamel tissue from freeze-fractured pig third molar were treated with antiserum against recombinant mouse amelogenin and immunoreactivity confirmed by Western blot analysis. The samples were then treated with the goat anti-rabbit IgG conjugated with 10-nm gold particles. The control samples were treated with the secondary antibody only. The in-lens secondary electrons detector and quadrant back-scattering detector were employed to reveal the high-resolution morphology of enamel structures and gold particle distribution. The immunolabeling showed a preference of the gold particle localization along the side faces of the ribbon-like apatite crystals. The preferential localization of amelogenin in vivo on enamel crystals strongly supports its direct function in controlling crystal morphology.
The Illness Attitude Scales
Authors: Sirri Laura , Grandi Silvana , Fava Giovanni A.
Published: August 2008

Background: The Illness Attitude Scales (IAS) were developed by Robert Kellner as a clinimetric index for measuring hypochondriacal fears and beliefs (worry about illness, concerns about pain, health habits, hypochondriacal beliefs, thanatophobia, disease phobia, bodily preoccupations, treatment experience and effects of symptoms). The IAS have been extensively used in the past two decades, but there has been no comprehensive review of their properties and applications. Methods: A review of the literature using both computerized (Medline, PsycINFO) and manual searches was performed. Results: The IAS were found to successfully discriminate between hypochondriacal patients and control subjects, and between patients with various manifestations of illness behaviour. They showed a high test-retest reliability in normal subjects, and changed in the expected direction after treatment of hypochondriasis. The IAS were also positively related to other hypochondriasis-related measures, and yielded important information in a variety of medical and surgical settings. Their content has paved the way for the development of some of the Diagnostic Criteria for Psychosomatic Research. Conclusions: The clinimetric properties and high sensitivity of the IAS make them the gold standard for the self-rated assessment of hypochondriacal fears and beliefs.
Identifying Patients at Risk of Complete Nonresponse in the Outpatient Treatment of Depression
Authors: Van Henricus L. , Dekker Jack , Peen Jaap , van Aalst Gerda , Schoevers Robert A.
Published: August 2008

Background: Although complete nonresponse in depression treatment is considered to be a major problem in clinical practice, research in this area is very limited. The objective of this preliminary study was to determine the frequency and predictors of complete nonresponse in different treatments for depression. Methods: Post hocanalysis of the pooled data of 3 consecutive randomized controlled trials of outpatient depression treatment was conducted. The subjects were 313 patients with major depressive disorder and 17-item Hamilton Rating Scale for Depression (HAM-D-17) scores between 14 and 25 who were treated for 6 months with either pharmacotherapy, short-term psychodynamic supportive psychotherapy or combined therapy. Complete nonresponse was defined as a <25% response according to the HAM-D-17. Sociodemographic factors, depression features and adherence were investigated as predictors in a multivariate stepwise logistic regression analysis. Results: Overall, nonresponse occurred in 34% of the patients. In pharmacotherapy this was 46%, in psychotherapy 39% and in combined therapy 28%. The severity of somatic symptoms was associated with nonresponse in both combined therapy and psychotherapy. No predictive factors were found in the case of pharmacotherapy. In psychotherapy, nonresponse was related to age above 40 years, chronic depression and nonadherence by the patient. In the case of combined therapy, younger age, previous use of an antidepressant and having a previous depressive episode were associated with nonresponse. Conclusion: Easily measurable patient characteristics may help to identify patients at risk of complete nonresponse to treatment. It is suggested that predictors may differ across treatment modalities. However, head-to-head comparisons are required before it can be recommended to take this into account when selecting the most appropriate treatment for individual depressed patients.
Hyperoxaluria Is Reduced and Nephrocalcinosis Prevented with an Oxalate-Degrading Enzyme in Mice with Hyperoxaluria
Authors: Grujic Danica , Salido Eduardo C. , Shenoy Bhami C. , Langman Craig B. , McGrath Margaret E. , Patel Reena J. , Rashid Aftab , Mandapati Saraswathi , Jung Chu W. , Margolin Alexey L.
Published: August 2008

Background/Aims: Hyperoxaluria is a major risk factor for recurrent urolithiasis and nephrocalcinosis. We tested an oral therapy with a crystalline, cross-linked formulation of oxalate-decarboxylase (OxDc-CLEC®) on the reduction of urinary oxalate and decrease in the severity of kidney injury in two models: AGT1 knockout mice (AGT1KO) in which hyperoxaluria is the result of an Agxt gene deficiency, and in AGT1KO mice challenged with ethylene glycol (EG). Methods: Four different doses of OxDc-CLEC mixed with the food, or placebo were given to AGT1KO mice (200 mg/day, n = 7) for 16 days and to EG-AGT1KO mice (5, 25, and 80 mg, n = 11) for 32 days. Results: Oral therapy with 200 mg OxDc-CLEC reduced both urinary (44%) and fecal oxalate (72%) in AGT1KO mice when compared to controls. Similarly, in EG-AGT1KO mice, each of the three doses of OxDc-CLEC produced a 30–50% reduction in hyperoxaluria. A sustained urinary oxalate reduction of 40% or more in the 80 mg group led to 100% animal survival and complete prevention of nephrocalcinosis and urolithiasis. Conclusion: These data suggest that oral therapy with OxDc-CLEC may reduce hyperoxaluria, prevent calcium oxalate nephrocalcinosis and urolithiasis, and can represent a realistic option for the treatment of human hyperoxaluria, independent of cause.
Decreased Parietal Cortex Activity during Mental Rotation in Children with Congenital Hypothyroidism
Authors: Blasi Valeria , Longaretti Roberta , Giovanettoni Chiara , Baldoli Cristina , Pontesilli Silvia , Vigone Cristina , Saccuman Cristina , Nigro Francesco , Chiumello Giuseppe , Scotti Giuseppe , Weber Giovanna
Published: August 2008

Background: The ability to detect the spatial characteristics of objects and to rotate them mentally is frequently impaired in early treated congenital hypothyroidism (CH) children. Aims: To explore the neural substrate of the visuospatial difficulty in children with CH, we studied 15 children with CH (8–10 years) and 13 age-matched control children with functional magnetic resonance imaging (fMRI) using a mental rotation task (VST). Results: Performance at VST was significantly different between the two groups. Moreover, fMRI data showed greater activation in the superior parietal cortex in control children while children with CH had greater activation in the bilateral SMA and the opercular region of the precentral gyrus, the adjacent insula and the left somatosensory parietal cortex. Furthermore, children with CH deactivated the inferior parietal cortex (Brodmann area 40) more than controls. Conclusion: We suggest that the poorer performance of children with CH on VST task is related to the decreased activation in brain areas important for the mental representation of the objects’ spatial characteristics, with increased recruitment of regions involved in the representation of somatosensory whole-body information. More studies will be necessary to understand if this different effectiveness in VST reflects immaturity of the neural system or its actual impairment.
Isolation of Distinct Progenitor Stem Cell Populations from Dental Pulp
Authors: Waddington Rachel J. , Youde Sarah J. , Lee Chi P. , Sloan Alastair J.
Published: August 2008

The present study compared the cellular characteristics of progenitor stem cell populations present in adult dental pulp, isolated by different methods utilizing 2 different features of stem cell biology. One population expressing high levels of β1 integrin was isolated by preferential selection of adherent cells to fibronectin over 20 min. In an alternative approach, cells expressing the embryonic neural crest cell marker, low-affinity nerve growth factor receptor (LANGFR), were selected by magnetic-activated cell sorting. For each method, clonal cell lines were established and expanded in culture. One clone derived via the respective methods was examined for embryonic/progenitor cell markers by immunocytochemistry and RT-PCR. Both clonal populations demonstrated the expression of stro-1 and stained positive for vimentin, demonstrating mesenchymal lineage. Of note, cells selected for LANGFR cells demonstrated the additional expression of CD105 and Notch 2. For both clonal populations, expanded cultures demonstrated the ability to differentiate into osteoblasts, adipocytes and chondrocytes. These results would suggest the potential isolation of 2 progenitor cell populations exhibiting different cellular characteristics in terms of their embryonic nature. The potential for both cell populations to derive from a common origin is discussed.
Identification and Functional Analysis of Two Novel PAX9 Mutations
Authors: Wang Ying , Wu Hua , Wu Jingfeng , Zhao Hongshan , Zhang Xiaoxia , Mues Gabriele , D’Souza Rena N. , Feng Hailan , Kapadia Hitesh
Published: August 2008

The paired-domain transcription factor PAX9 plays a critical role in tooth development, as heterozygous mutations in PAX9 have been shown to be associated with human tooth agenesis. In this study, we report 2 novel missense mutations, gly6arg (G6R) and ser43lys (S43K), in the paired domain of PAX9 in Chinese patients with varying degrees of nonsyndromic tooth agenesis. Excluding third molars, the individual with the G6R mutation was missing 2 mandibular incisors and a maxillary premolar, while the phenotype of individuals with the S43K mutation consisted of peg-shaped upper lateral incisors and missing molars, premolars and canines. As these 2 mutations occur at highly conserved amino acids in the PAX gene family and between different species, we further analyzed the effects of the mutations on the function of the resulting proteins. Immunofluorescence and immunoblotting studies showed that the mutations did not alter nuclear localization in mammalian cells. Gel shift and super shift assays indicate that both mutant proteins bound DNA at a lower level than the normal protein, with G6R having a greater affinity for DNA than S43K. Likewise, the G6R protein was able to transcriptionally activate a Bmp4 promoter construct to a greater extent than S43K. Our finding that the severity of tooth agenesis in the patients was correlated to the DNA-binding capacity of the mutated PAX9 9proteins supports the hypothesis that DNA binding is responsible for the genetic defect.
Molecular Switches Involving Homeodomain Proteins, HOXA10 and RUNX2 Regulate Osteoblastogenesis
Authors: Hassan Mohammad Q. , Saini Sharanjot , Gordon Jonathan A.R. , van Wijnen Andre J. , Montecino Martin , Stein Janet L. , Stein Gary S. , Lian Jane B.
Published: August 2008

The osteoinductive BMP2 signal facilitates commitment to the osteoblast phenotype by inducing several classes of early response genes. Among these are bone-related HOX factors, homeodomain, RUNX and OSTERIX proteins. Here we demonstrate molecular events among BMP2-induced transcription factors that constitute a network of molecular switches on promoters of bone-related genes to coordinate their temporal expression during cellular differentiation. Our studies provide evidence for (1) selective association of HOXA10, MSX2, DLX3 and DLX5 homeodomain transcription factors on Runx2 and OC genes at stages of osteoblast maturation as well as (2) participation of these factors with RUNX2 in chromatin remodeling of bone-specific genes for repression, activation and attenuation of transcription. These findings reveal the requirement for multiple levels of control for the appropriate timing of osteoblast-related gene expression.
Interspecies differences in coagulation profile
Authors: Siller-Matula Jolanta M. , Plasenzotti Roberto , Spiel Alexander , Quehenberger Peter , Jilma Bernd
Published: August 2008

Many animals are used in research on blood coagulation and fibrinolysis, but the relevance of animal models to human health is often questioned because of differences between species. The objective was to find an appropriate animal species
,
which mimics the coagulation profile in humans most adequately. Species differences in the coagulation profile with and without thrombin stimulation in vitro were assessed in whole blood by Rotation Thromboelastometry (ROTEM). Endogenous thrombin generation was measured in platelet-poor plasma. Measurements were performed in blood from five different species: humans, rats, pigs, sheep and rabbits. In humans and sheep, the clotting time (ROTEM) was in the same range with or without thrombin stimulation and a 100-fold lower dose of thrombin (0.002 IU) was required to cause a shortening in the clotting time as compared to rats, pigs and rabbits (0.2 IU) (p<0.05).Similarly, the endogenous thrombin potential (ETP) was in the same range in humans and sheep. The maximum clot firmness with or without thrombin stimulation was similar in rabbits and humans. The maximum lysis with or without thrombin stimulation was similar in humans and pigs. Significant species differences exist in the coagulation profile with or without thrombin stimulation. Most importantly, sheep had a clotting time most similar to humans and could thus be a suitable species for translational coagulation studies. Moreover, our findings confirm the potential usefulness of pigs as an experimental species to study fibrinolytic pathway and support the usefulness of rabbits as a species for examining platelets.
Predicting recurrences or major bleeding in cancer patients with venous thromboembolism
Authors: Trujillo-Santos Javier , Nieto José , Tiberio Gregorio , Piccioli Andrea , Micco Pierpaolo , Prandoni Paolo , Monreal Manuel
Published: August 2008

Cancer patients with acute venous thromboembolism (VTE) have an increased incidence of recurrences and bleeding complications while on anticoagulant therapy. Methods RIETE is an ongoing registry of consecutive patients with acute VTE. We tried to identify which cancer patients are at a higher risk for recurrent pulmonary embolism (PE), deep vein thrombosis (DVT) or major bleeding. Up to May 2007, 3, 805 cancer patients had been enrolled in RIETE. During the first three months of follow-up after the acute, index VTE event, 90 (2.4%) patients developed recurrent PE, 100 (2.6%) recurrent DVT, 156 (4.1%) had major bleeding. Forty patients (44%) died of the recurrent PE,46 (29%) of bleeding. On multivariate analysis, patients aged <65 years (odds ratio [OR]: 3.0; 95% confidence interval [CI]: 1.9–4.9), with PE at entry (OR: 1.9; 95% CI: 1.2–3.1), or with <3 months from cancer diagnosis to VTE (OR: 2.0; 95% CI: 1.2–3.2) had an increased incidence of recurrent PE. Those aged <65 years (OR: 1.6; 95% CI: 1.0–2.4) or with <3 months from cancer diagnosis (OR: 2.4; 95% CI: 1.5–3.6) had an increased incidence of recurrent DVT. Finally, patients with immobility (OR: 1.8; 95% CI: 1.2–2.7), metastases (OR: 1.6; 95% CI: 1.1–2.3), recent bleeding (OR: 2.4; 95% CI: 1.1–5.1), or with creatinine clearance <30 ml/ min (OR: 2.2; 95% CI: 1.5–3.4), had an increased incidence of major bleeding. With some variables available at entry we may identify those cancer patients withVTE at a higher risk for recurrences or major bleeding.
Evaluation of liposomal dose in recombinant factor VIII reconstituted with pegylated liposomes for the treatment of patients with severe haemophilia A
Authors: Plyushch Olga P. , Andreeva Tatyana A. , Khametova Rosa N. , Spira Jack
Published: August 2008

Patients with haemophilia A treated prophylactically require frequent factor VIII (FVIII) infusions for bleed protection. Recombinant sucrose-formulated FVIII (rFVIII-FS) with pegylated liposomes (PEGLip-rFVIII-FS) was previously shown to extend the bleed-free period after prophylactic infusion versus rFVIII-FS using two doses of FVIII with a fixed amount of liposomal diluent. This randomised, subject-blinded, four-way crossover study evaluated the efficacy and safety of PEGLip-rFVIII-FS using various quantities of pegylated liposomes with a fixed FVIII dose. Adults with severe haemophilia A were randomised to one of four treatment arms. Each arm had four treatment segments, with each segment consisting of a prophylactic infusion followed by on-demand infusions. The prophylactic infusions used a fixed dose of 35 IU/kg rFVIII-FS, but varied in the amount of pegylated liposomes used for reconstitution (4.2, 12.6, or 22.1 mg/kg of body weight or water as a control). On-demand infusions all used 35 IU/kg rFVIII-FS. After treatment of spontaneous bleeds and a wash-out, subjects crossed to another treatment segment (i.e. another PEGLip-rFVIII-FS dose or control).Sixteen subjects enrolled in and completed the study. Mean number of bleed-free days after prophylactic infusion increased from 7.8 days for control rFVIII-FS to 8.7, 10.8, and 10.9 days for PEGLip-rFVIII-FS reconstituted in 4.2, 12.6, and 22.1 mg/kg of pegylated liposomes, respectively. The difference in bleed-free days approached but did not achieve statistical significance in this small study population. No drug-related adverse events or inhibitors were reported. This study helps establish the optimal concentration of liposomes in PEGLip-rFVIII-FS to prolong the post-infusion bleed-free period.
Heparin-induced antibodies and cardiovascular risk in patients on dialysis
Authors: Segal Jodi B. , Plantinga Laura C. , Fink Nancy E. , Kerman Jonathan S. , Kickler Thomas S. , Coresh Josef , Gardner Lawrence B. , Asmis Lars M.
Published: August 2008

The clinical relevance of heparin-induced antibodies (HIA) in the absence of thrombocytopenia remains to be defined. The aims of this study were (i) to determine the prevalence of HIA in patients treated by dialysis, (ii) to determine the prevalence of thrombocytopenia and heparin-induced thrombocytopenia (HIT), and (iii) to test whether HIA are associated with adverse outcomes. Sera from 740 patients treated by hemodialysis (HD, n=596) and peritoneal dialysis (PD, n=144) were tested for HIA (IgG, IgA or IgM) by masked investigators at approximately six months after enrolment in the Choices for Healthy Outcomes in Caring for End-Stage Renal Disease (CHOICE) study. We assessed, with time-to-event Cox proportional hazards models, whether the presence of HIA predicted any of four clinical outcomes: arterial cardiovascular events, venous thromboembolism, vascular access occlusion and mortality. HIA prevalence was 10.3% overall. HIA positivity did not predict development of thrombocytopenia or any of the four clinical outcomes over a mean follow-up of 3.6 years, with hazard ratios for arterial cardiovascular events of 0.98 (95% confidence interval 0.70–1.37), venous thromboembolism 1.39 (0.17–11.5), vascular access occlusion 0.82 (0.40–1.71), and mortality 1.18 (0.85–1.64). Chronic intermittent heparin exposure was associated with a high seroprevalence of HIA. In dialysis patients these antibodies were not an independent risk factor for cardiovascular events and mortality. Our data do not suggest that dialysis patients should be monitored for HIA antibodies in the absence of thrombocytopenia.
Serpin-independent anticoagulant activity of a fucosylated chondroitin sulfate
Authors: Glauser Bianca F. , Pereira Mariana S. , Monteiro Robson Q. , Mourão Paulo A. S.
Published: August 2008

Fucosylated chondroitin sulfate is a glycosaminoglycan from sea cucumber composed of a chondroitin sulfate-like core with branches of sulfated fucose. This glycosaminoglycan has high anticoagulant and antithrombotic activities. Its serpin-dependent anticoagulant activity is mostly due to activating thrombin inhibition by heparin cofactor II. Here, we evaluated the anticoagulant activity of fucosylated chondroitin sulfate using antithrom-bin- and heparin cofactor II-free plasmas. In contrast to mammalian heparin, the invertebrate glycosaminoglycan is still able to prolong coagulation time and delay thrombin and factor Xa generation in serpin-free plasmas. These observations suggest that fucosylated chondroitin sulfate has a serpin-independent anticoagulant effect. We further investigated this effect using purified blood coagulation proteins. Clearly, fucosylated chondroitin sulfate inhibits the intrinsic tenase and prothrombinase complexes, which are critical for thrombin generation. It is possible

that the invertebrate chondroitin sulfate inhibits interactions between cofactor Va and factor Xa. We also employed chemically modified polysaccharides in order to trace a structure versus activity relationship. Removal of the sulfated fucose branches, but not reduction of the glucuronic acid residues to glucose, abolished its activity. In conclusion, fucosylated chondroitin sulfate has broader effects on the coagulation system than mammalian glycosaminoglycans. In addition to its serpin-dependent inhibition of coagulation protease, it also inhibits the generation of factor Xa and thrombin by the tenase and prothrombinase complexes, respectively. In plasma systems, the serpin-independent anticoagulant effect of fucosylated chondroitin sulfate predominates over its serpin-dependent action. This glycosaminoglycan opens new avenues for the development of antithrombotic agents.
Simplified recombinant plasmin: Production and functional comparison of a novel thrombolytic molecule with plasma-derived plasmin
Authors: Hunt Jennifer , Petteway Stephen , Scuderi Philip , Novokhatny Valery
Published: August 2008

A simplified and fully functional deletion mutant of plasminogen was created in which the middle portion of the molecule was removed, resulting in kringle 1 attachment to the serine protease domain. This recombinant plasminogen deletion mutant, Δ(K2-K5)Pg, was produced in the form of inclusion bodies at the yield of up to 200 mg/l in an Escherichia coliT7 expression system. Following protein refolding and purification on lysine-Sepharose, the conversion of the recombinant molecule Δ(K2-K5)Pg to the active enzyme mutant Δ(K2-K5)Pm by plasminogen activators was evaluated, and functional characteristics of the simplified plasmin were studied. Properties of Δ(K2-K5)Pg were similar to native, human plasma-derived plasminogen. Δ(K2-K5)Pg effectively bound ε-aminocaproic acid (Kd = 1 1.3 ± 2.3 μ. M) and fibrin (C50 ≈ 0.3 μM).The plasminogen activators streptokinase, urokinase, and tissue plasminogen activator effectively converted the recombinant zymogen Δ(K2-K5)Pg to the active recombinant enzyme, Δ(K2-K5)Pm. Additionally, Δ[K2-K5]Pm was rapidly inhibited by α2-antiplas-min(1.1±0.1x107M−1s−1)andα2-macroglobulin(7.6±0.6x105 M−1s−1). In an in-vitro model, Δ(K2-K5)Pm demonstrated fibrinolytic potency comparable to human plasma-derived plasmin. Because of their unique biochemistry, including fibrin-binding properties and rapid inhibition by α2-antiplasmin, both native plasmin and a simplified deletion mutant of plasmin are potentially safe and effective direct thrombolytic agents for various thrombotic conditions. Further studies evaluating the in-vivo pharmacologic safety and clinical efficacy of this simplified plasmin (i.e. Δ[K2-K5]Pm) are warranted.
Heparan sulfate proteoglycan is essential to thrombin-induced calcium transients and nitric oxide production in aortic endothelial cells
Authors: Kimura Chiwaka , Oike Masahiro
Published: August 2008

Thrombin induces Ca2+ transients and subsequent nitric oxide (NO) production in vascular endothelial cells. Thrombin cleaves protease-activated receptors, resulting in activation of intracellular signals, but it is not clarified how the extracellular thrombin stays around the cells to exert its enzyme activities. This study aimed to investigate the possible involvement of heparin sulfate proteoglycan (HSPG) in the effects of thrombin on vascular endothelium. Heparinase III completely removed the polysaccharide chain of HSPG in bovine aortic endothelial cells (BAECs).Thrombin induced Ca2+ transients in control BAECs, but not in heparinase III-treated BAECs. In contrast, ATP induced Ca2+ transients both in control and heparinase III-treated BAECs. Thrombin that was pre-incubated with heparin also failed to induced Ca2+ transients in BAECs. Furthermore, thrombin-induced NO production, as assessed with DAF-2 flu-orescence, was suppressed in heparinase III-treated BAECs and by the pre-incubation of thrombin with heparin. ATP-induced NO production was, however, not affected in heparinase III-treated BAECs. These results indicate that it is essential for thrombin to bind to the polysaccharide chain of HSPG for inducing Ca2+ transients and NO production in BAECs.
Crosstalk Between Autophagy and Apoptosis in Heart Disease
Authors: Nishida Kazuhiko , Yamaguchi Osamu , Otsu Kinya
Published: August 2008

Autophagy is a cell survival mechanism that involves degradation and recycling of cytoplasmic components, such as long-lived proteins and organelles. In addition, autophagy mediates cell death under specific circumstances. Apoptosis, a form of programmed cell death, has been well characterized, and the molecular events involved in apoptotic death are well understood. Damaged cardiomyocytes that show characteristics of autophagy have been observed during heart failure. However, it remains unclear whether autophagy is a sign of failed cardiomyocyte repair or is a suicide pathway for the failing cardiomyocytes. Although autophagy and apoptosis are markedly different processes, several pathways regulate both autophagic and apoptotic machinery and autophagy can cooperate with apoptosis. This review summarizes the evidence for crosstalk between autophagy and apoptosis.
Neural Correlates of Subjective Awareness and Unconscious Processing: An ERP Study
Authors: Lamy Dominique , Salti Moti , Bar-Haim Yair
Published: August 2008

The aim of the present study was to dissociate the ERP (Event Related Potentials) correlates of subjective awareness from those of unconscious perception. In a backward masking paradigm, participants first produced a forced-choice response to the location of a liminal target presented for an individually calibrated duration, and then reported on their subjective awareness of the target's presence. We recorded (Event-Related Potentials) ERPs and compared the ERP waves when observers reported being aware vs. unaware of the target but localized it correctly, thereby isolating the neural correlates of subjective awareness while controlling for differences in objective performance. In addition, we compared the ERPs when participants were subjectively unaware of the target's presence and localized it correctly versus incorrectly, thereby isolating the neural correlates of unconscious perception. All conditions involved stimuli that were physically identical and were presented for the same duration. Both behavioral measures were associated with modulation of the amplitude of the P3 component of the ERP. Importantly, this modulation was widely spread across all scalp locations for subjective awareness, but was restricted to the parietal electrodes for unconscious perception. These results indicate that liminal stimuli that do not affect performance undergo considerable processing and that subjective awareness is associated with a late wave of activation with widely distributed topography.
Contribution of Prior Semantic Knowledge to New Episodic Learning in Amnesia
Authors: Kan Irene P. , Alexander Michael P. , Verfaellie Mieke
Published: August 2008

We evaluated whether prior semantic knowledge would enhance episodic learning in amnesia. Subjects studied prices that are either congruent or incongruent with prior price knowledge for grocery and household items and then performed a forced-choice recognition test for the studied prices. Consistent with a previous report, healthy controls' performance was enhanced by price knowledge congruency; however, only a subset of amnesic patients experienced the same benefit. Whereas patients with relatively intact semantic systems, as measured by an anatomical measure (i.e., lesion involvement of anterior and lateral temporal lobes), experienced a significant congruency benefit, patients with compromised semantic systems did not experience a congruency benefit. Our findings suggest that when prior knowledge structures are intact, they can support acquisition of new episodic information by providing frameworks into which such information can be incorporated.
Task-relevant Output Signals are Sent from Monkey Dorsolateral Prefrontal Cortex to the Superior Colliculus during a Visuospatial Working Memory Task
Authors: Johnston Kevin , Everling Stefan
Published: August 2008

Visuospatial working memory is one of the most extensively investigated functions of the dorsolateral prefrontal cortex (DLPFC). Theories of prefrontal cortical function have suggested that this area exerts cognitive control by modulating the activity of structures to which it is connected. Here, we used the oculomotor system as a model in which to characterize the output signals sent from the DLPFC to a target structure during a classical spatial working memory task. We recorded the activity of identified DLPFC–superior colliculus (SC) projection neurons while monkeys performed a memory-guided saccade task in which they were required to generate saccades toward remembered stimulus locations. DLPFC neurons sent signals related to all aspects of the task to the SC, some of which were spatially tuned. These data provide the first direct evidence that the DLPFC sends task-relevant information to the SC during a spatial working memory task, and further support a role for the DLPFC in the direct modulation of other brain areas.
Don't Do It! Cortical Inhibition and Self-attribution during Action Observation
Authors: Schütz-Bosbach Simone , Avenanti Alessio , Aglioti Salvatore Maria , Haggard Patrick
Published: August 2008

Numerous studies suggest that both self-generated and observed actions of others activate overlapping neural networks, implying a shared, agent-neutral representation of self and other. Contrary to the shared representation hypothesis, we recently showed that the human motor system is not neutral with respect to the agent of an observed action [Schütz-Bosbach, S., Mancini, B., Aglioti, S. M., & Haggard, P. Self and other in the human motor system. Current Biology, 16, 1830–1834, 2006]. Observation of actions attributed to another agent facilitated the motor system, whereas observation of identical actions linked to the self did not. Here we investigate whether the absence of motor facilitation for observing one's own actions reflects a specific process of cortical inhibition associated with self-representation. We analyzed the duration of the silent period induced by transcranial magnetic stimulation of the motor cortex in active muscles as an indicator of motor inhibition. We manipulated whether an observed action was attributed to another agent, or to the subjects themselves, using a manipulation of body ownership on the basis of the rubber hand illusion. Observation of actions linked to the self led to longer silent periods than observation of a static hand, but the opposite effect occurred when observing identical actions attributed to another agent. This finding suggests a specific inhibition of the motor system associated with self-representation. Cortical suppression for actions linked to the self might prevent inappropriate perseveration within the motor system.
Posterior Beta and Anterior Gamma Oscillations Predict Cognitive Insight
Authors: Sheth Bhavin R. , Sandkühler Simone , Bhattacharya Joydeep
Published: August 2008

Pioneering neuroimaging studies on insight have revealed neural correlates of the emotional “Aha!” component of the insight process, but neural substrates of the cognitive component, such as problem restructuring (a key to transformative reasoning), remain a mystery. Here, multivariate electroencephalogram signals were recorded from human participants while they solved verbal puzzles that could create a small-scale experience of cognitive insight. Individuals responded as soon as they reached a solution and provided a rating of subjective insight. For unsolved puzzles, hints were provided after 60 to 90 sec. Spatio-temporal signatures of brain oscillations were analyzed using Morlet wavelet transform followed by exploratory parallel-factor analysis. A consistent reduction in beta power (15–25 Hz) was found over the parieto-occipital and centro-temporal electrode regions on all four conditions—(a) correct (vs. incorrect) solutions, (b) solutions without (vs. with) external hint, (c) successful (vs. unsuccessful) utilization of the external hint, and d) self-reported high (vs. low) insight. Gamma band (30–70 Hz) power was increased in right fronto-central and frontal electrode regions for conditions (a) and (c). The effects occurred several (up to 8) seconds before the behavioral response. Our findings indicate that insight is represented by distinct spectral, spatial, and temporal patterns of neural activity related to presolution cognitive processes that are intrinsic to the problem itself but not exclusively to one's subjective assessment of insight.