bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of Cell Death in Spider Embryos Using TUNEL
Authors: Prpic Nikola-Michael , Schoppmeier Michael , Damen Wim G.M. 
Published: October 2008

INTRODUCTIONThe spider Cupiennius salei, commonly known as the American Wandering Spider, is a particularly useful laboratory model for embryological studies because of the availability of tools to study and manipulate its embryonic development. Cupiennius is used to study axis formation, segmentation, appendage development, neurogenesis, and silk production. These studies contribute to our understanding of the evolution of these processes, but they also help us to understand the origin and diversification of evolutionary novelties. Comparisons between spiders and insects can show the degree of conservation and divergence of developmental mechanisms during arthropod evolution. Any embryological feature conserved between spiders and insects is likely to represent an ancestral feature for arthropods. Comparative molecular embryological work in insects and spiders should eventually allow us to define a molecular archetype for the phylum Arthropoda. This in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. A feature of apoptosis (i.e., cell death) is the cleavage or fragmentation of DNA that occurs in dead or dying cells. This protocol describes the detection of fragmented DNA in whole-mount Cupiennius embryos. The 3′-OH ends of these DNA fragments can be labeled with the terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) technique. This protocol uses a terminal deoxynucleotidyl transferase to add labeled dUTP to the fragmented DNA, and this label is then detected by immunocytochemistry. The TUNEL technique is a relatively easy way to obtain a reliable picture of the cell death pattern during normal and abnormal development.
Basic Maintenance and Breeding of the Opossum Monodelphis domestica
Authors: Keyte Anna L. , Smith Kathleen K. 
Published: October 2008

INTRODUCTIONMonodelphis domestica, the gray, short-tailed, or laboratory opossum, is the most commonly used laboratory marsupial. In addition to the factors that make it a convenient laboratory animal (small size, ease of care, nonseasonal breeding), it is the first marsupial whose genome has been sequenced. Monodelphis has proven useful as a model organism for studies on spinal cord regeneration, ultraviolet (UV)-induced melanoma, and genetic influences on cholesterol, as well as comparative studies of the immune system. In addition, Monodelphis has been used to understand the basic functions of the olfactory system and the role of various olfactory chemicals in social and reproductive behavior. Recently, Monodelphis has been used to understand fundamental aspects of marsupial development, anatomy, evolution, and evolutionary consequences of the derived marsupial mode of development and reproduction. Monodelphis are easily maintained and bred in the lab. To do extensive embryonic work, a reasonably large breeding colony must be maintained. A colony of ~100 animals (~3:1 female:male ratio) allows for sacrifice of up to 12 pregnant females per month for experimental purposes, as well as for replenishment of the colony. However, because adults will fight and often kill one another if kept in the same cage for prolonged periods, we have developed a special breeding protocol that provides high rates of breeding success (75%-90%), with minimal injury due to fighting. Here, we outline this breeding strategy and describe how to successfully maintain a colony of Monodelphis in a laboratory setting.
Cell Membrane Surface Potential (ψ
0) Plays a Dominant Role in the Phytotoxicity of Copper and Arsenate
Authors: Wang Peng , Zhou Dongmei , Kinraide Thomas B. , Luo Xiaosan , Li Lianzhen , Li Dandan , Zhang Hailin 
Published: October 2008

Abstract
Negative charges at cell membrane surfaces (CMS) create a surface electrical potential (ψ 0) that affects ion concentrations at the CMS and consequently affects the phytotoxicity of metallic cations and metalloid anions in different ways. The ζ potentials of root protoplasts of wheat (Triticum aestivum), as affected by the ionic environment of the solution, were measured and compared with the values of ψ 0 calculated with a Gouy-Chapman-Stern model. The mechanisms for the effects of cations (H+, Ca2+, Mg2+, Na+, and K+) on the acute toxicity of Cu2+ and As(V) to wheat were studied in terms of ψ 0. The order of effectiveness of the ions in reducing the negativity of ψ 0 was H+ &gt; Ca2+ ≈ Mg2+ &gt; Na+ ≈ K+. The calculated values of ψ 0 were proportional to the measured ζ potentials (r 2 = 0.93). Increasing Ca2+ or Mg2+ activities in bulk-phase media resulted in decreased CMS activities of Cu2+ ({Cu2+}0) and increased CMS activities of As(V) ({As(V)}0). The 48-h EA50{Cu2+}b ({Cu2+} in bulk-phase media accounting for 50% inhibition of root elongation over 48 h) increased initially and then declined, whereas the 48-h EA50{As(V)}b decreased linearly. However, the intrinsic toxicity of Cu2+ (toxicity expressed in terms of {Cu2+}0) appeared to be enhanced as ψ 0 became less negative and the intrinsic toxicity of As(V) appeared to be reduced. The ψ 0 effects, rather than site-specific competitions among ions at the CMS (invoked by the biotic ligand model), may play the dominant role in the phytotoxicities of Cu2+ and As(V) to wheat.
Arabidopsis Transcriptome Reveals Control Circuits Regulating Redox Homeostasis and the Role of an AP2 Transcription Factor
Authors: Khandelwal Abha , Elvitigala Thanura , Ghosh Bijoy , Quatrano Ralph S. 
Published: October 2008

Abstract
Sensors and regulatory circuits that maintain redox homeostasis play a central role in adjusting plant metabolism and development to changing environmental conditions. We report here control networks in Arabidopsis (Arabidopsis thaliana) that respond to photosynthetic stress. We independently subjected Arabidopsis leaves to two commonly used photosystem II inhibitors: high light (HL) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Microarray analysis of expression patterns during the period of redox adjustment to these inhibitors reveals that 20% and 8% of the transcriptome are under HL and DCMU regulation, respectively. Approximately 6% comprise a subset of genes common to both perturbations, the redox responsive genes (RRGs). A redox network was generated in an attempt to identify genes whose expression is tightly coordinated during adjustment to homeostasis, using expression of these RRGs under HL conditions. Ten subnetworks were identified from the network. Hierarchal subclustering of subnetworks responding to the DCMU stress identified novel groups of genes that were tightly controlled while adjusting to homeostasis. Upstream analysis of the promoters of the genes in these clusters revealed different motifs for each subnetwork, including motifs that were previously identified with responses to other stresses, such as light, dehydration, or abscisic acid. Functional categorization of RRGs demonstrated involvement of genes in many metabolic pathways, including several families of transcription factors, especially those in the AP2 family. Using a T-DNA insertion in one AP2 transcription factor (redox-responsive transcription factor 1 [RRTF1]) from the RRGs, we showed that the genes predicted to be within the subnetwork containing RRTF1 were changed in this insertion line (Δrrtf1). Furthermore, Δrrtf1 showed greater sensitivity to photosynthetic stress compared to the wild type.
Proteolysis-Independent Downregulation of DELLA Repression inArabidopsisby the Gibberellin Receptor GIBBERELLIN INSENSITIVE DWARF1
Authors: Ariizumi Tohru , Murase Kohji , Sun Tai-ping , Steber Camille M. 
Published: October 2008

AbstractThis article presents evidence that DELLA repression of gibberellin (GA) signaling is relieved both by proteolysis-dependent and -independent pathways in Arabidopsis thaliana. DELLA proteins are negative regulators of GA responses, including seed germination, stem elongation, and fertility. GA stimulates GA responses by causing DELLA repressor degradation via the ubiquitin-proteasome pathway. DELLA degradation requires GA biosynthesis, three functionally redundant GA receptors GIBBERELLIN INSENSITIVE DWARF1 (GID1a, b, and c), and the SLEEPY1 (SLY1) F-box subunit of an SCF E3 ubiquitin ligase. The sly1 mutants accumulate more DELLA proteins but display less severe dwarf and germination phenotypes than the GA biosynthesis mutant ga1-3 or the gid1abc triple mutant. Interestingly, GID1 overexpression rescued the sly1 dwarf and infertility phenotypes without decreasing the accumulation of the DELLA protein REPRESSOR OF ga1-3. GID1 rescue of sly1 mutants was dependent on the level of GID1 protein, GA, and the presence of a functional DELLA motif. Since DELLA shows increasing interaction with GID1 with increasing GA levels, it appears that GA-bound GID1 can block DELLA repressor activity by direct protein–protein interaction with the DELLA domain. Thus, a SLY1-independent mechanism for GA signaling may function without DELLA degradation.
Record of sea‐level fall in tropical carbonates
Authors: Schlager Wolfgang , Warrlich Georg 
Published: October 2008

ABSTRACTStratigraphic forward modeling and comparison with published case studies have been used to determine the controls and stability domains of two conceptual models concerning relative sea‐level fall in carbonate sequence stratigraphy. In the standard model, deposition occurs principally during rise and stillstands of relative sea level; a continuous erosional unconformity develops during sea‐level fall. The falling‐stage systems tract model (FST) postulates significant deposition during sea‐level fall. Sedimentological principles, numerical models and published case studies of tropical carbonate sequences indicate that presence or absence of FST is not simply a function of the rate of sea‐level fall but depends on the balance of the rates of erosion, sea‐level fall and carbonate production, whereby the FST is favoured by high production, slow erosion and slow sea‐level fall. Case studies plotted in the parameter space spanned by these variables support the modeling results. The ranges of rates required for the FST in the modeling runs are common in the geologic record. Consequently, the FST can be expected to be more common in tropical carbonate rocks than published records, particularly seismic data, currently indicate.
ADAMTS13 cleavage efficiency is altered by mutagenic and, to a lesser extent, polymorphic sequence changes in the A1 and A2 domains of von Willebrand factor
Authors: Pruss Cynthia M. , Notley Colleen R. P. , Hegadorn Carol A. , O’Brien Lee A. , Lillicrap David 
Published: October 2008

SummaryThe multimeric plasma protein von Willebrand factor (VWF) is regulated in size by its protease, ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13). Y1605‐M1606 cleavage site mutations and single nucleotide polymorphisms (SNPs) in the VWF A1 and A2 domains were examined for alteration in ADAMTS13‐mediated cleavage of VWF. Recombinant human full‐length VWF (rVWF) was digested with recombinant human ADAMTS13 (rADAMTS13) using a dialysis membrane method with 1·5 mol/l urea, and analyzed via multimer migration distance. The glutathione‐S‐transferase (GST) and histidine‐tagged construct, E1554‐R1668 of VWF (VWF115) was assayed via enzyme‐linked immunosorbent assay: VWF115 was bound to anti‐GST coated plates, digested with rADAMTS13, and intact VWF115 detected via horseradish peroxidase‐labelled anti‐histidine tag antibody. All alterations examined in the Y1605‐M1606 cleavage site greatly reduced the cleavability of VWF by ADAMTS13 in the rVWF assay. Greatest cleavage resistance in both assays was observed in Y1605A/M1606A. In contrast, Y1605H and M1606L show a loss of cleavability only in the rVWF assay, suggesting that an aromatic ring at 1605 is critical for ADAMTS13 recognition. Additionally, under our rVWF assay conditions, the G1643S polymorphism showed increased cleavage, suggesting a Type 2A VWD phenotype, while D1472H, Q1571H and P1601T showed slightly decreased ADAMTS13 cleavage. Our two complementary assay conditions show that A‐domain changes in VWF alter ADAMTS13‐mediated proteolysis.
Bortezomib is associated with better health‐related quality of life than high‐dose dexamethasone in patients with relapsed multiple myeloma: results from the APEX study
Authors: Lee Stephanie J. , Richardson Paul G. , Sonneveld Pieter , Schuster Michael W. , Irwin David , San Miguel Jesús‐F. , Crawford Bruce , Massaro Joseph , Dhawan Ravinder , Gupta Sanjay , Anderson Kenneth C. 
Published: October 2008

SummaryHealth‐related quality of life (HRQL) was prospectively measured during the phase III APEX trial of bortezomib versus dexamethasone in relapsed multiple myeloma patients. The European Organisation for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire – Core (QLQ‐C30) and Functional Assessment of Cancer Therapy/Gynecologic Oncology Group–Neurotoxicity (NTX) side‐effects questionnaires were administered at baseline and every 6 weeks up to 42 weeks. Patients receiving bortezomib (1·3 mg/m2, days 1, 4, 8 and 11 for eight 3‐week cycles, then days 1, 8, 15 and 22 for three 5‐week cycles; n = 296) demonstrated significantly better mean Global Health Status over the study versus patients receiving dexamethasone (40 mg/d, days 1–4, 9–12, and 17–20 for four 5‐week cycles, then days 1–4 only for five 4‐week cycles; n = 302), plus significantly better physical health, role, cognitive, and emotional functioning scores, lower dyspnoea and sleep symptom scores, and better NTX questionnaire score, using multiple imputation to account for missing data. Results were similar using available‐data analyses. Sensitivity analyses suggested that improved HRQL with bortezomib is at least partially explained by improved survival. These results show that bortezomib was associated with significantly better multidimensional HRQL compared with dexamethasone, consistent with the better clinical outcomes seen with bortezomib.
Differential clot stabilising effects of rFVIIa and rFXIII‐A2 in whole blood from thrombocytopenic patients and healthy volunteers
Authors: Johansson Pär I. , Jacobsen Niels , Viuff Dorthe , Olsen Eva H. N. , Rojkjaer Rasmus , Andersen Søren , Petersen Lars C. , Kjalke Marianne 
Published: October 2008

SummaryThe haemostatic effect of recombinant activated factor VII (rFVIIa; NovoSeven®) in thrombocytopenic patients has been a matter of controversy. Haemostasis by rFVIIa occurs via FVIIa‐mediated thrombin generation in a platelet‐dependent manner and may therefore be suboptimal in patients without functional platelets. Under such conditions, a clot‐stabilizing agent, such as factor XIII (FXIII), may supplement the effect of rFVIIa and improve haemostasis. Recombinant factor XIII (rFXIII‐A2) is produced as an A2 homodimer of the FXIII A subunit and is equivalent to cellular FXIII normally found in platelets. The combined effects of rFVIIa and rFXIII‐A2 were evaluated in clot lysis assays using factor XIII‐deficient plasma and by whole blood thrombelastography (TEG) analysis from normal donors and thrombocytopenic stem cell transplantation patients. Clotting time was shortened by rFVIIa (0·6–10 μg/ml). rFVIIa only modestly improved anti‐fibrinolysis, whereas rFXIII‐A2 (0–20 μg/ml) enhanced anti‐fibrinolysis without effect on clotting time. TEG analysis showed rFVIIa shortened the clotting time, and enhanced clot development, maximal mechanical strength and resistance to fibrinolysis, whereas, rFXIII‐A2 enhanced clot development, maximal mechanical strength and markedly enhanced resistance to fibrinolysis. These data illustrate that rFVIIa and rFXIII‐A2 contribute to clot formation and stability by different mechanisms suggesting enhanced haemostatic efficacy by combining these agents.
MicroRNA 155 modulates megakaryopoiesis at progenitor and precursor level by targeting Ets‐1 and Meis1 transcription factors
Authors: Romania Paolo , Lulli Valentina , Pelosi Elvira , Biffoni Mauro , Peschle Cesare , Marziali Giovanna 
Published: October 2008

SummaryMicroRNAs (miRNAs) control basic biological functions and are emerging as key regulators of haematopoiesis. This study focused on the functional role of MIRN155 on megakaryocytic (MK) differentiation of human cord blood CD34+ haematopoietic progenitor cells (HPCs). MIRN155, abundantly expressed in early HPCs, decreases sharply during MK differentiation. Functional studies showed that enforced expression of MIRN155 impairs proliferation and differentiation of MK cells. Furthermore, HPCs transfected with MIRN155 showed a significant reduction of their MK clonogenic capacity, suggesting that down‐modulation of this miRNA favours MK progenitor differentiation. Consistent with this observation, MIRN155 down‐regulates, by directly binding to their 3′‐UTR, the expression of Ets‐1 and Meis1, two transcription factors with well‐known functions in MK cells. These results show that the decline of MIRN155 is required for MK proliferation and differentiation at progenitors and precursors level and indicate that sustained expression of MIRN155 inhibits megakaryopoiesis.
Thrombin activatable fibrinolysis inhibitor gene polymorphisms are associated with antigenic levels in the Asian‐Indian population but may not be a risk for stroke
Authors: Biswas Arijit , Tiwari Arun K. , Ranjan Ravi , Meena Arvind , Akhter Mohammad S. , Yadav Birendra K. , Behari Madhuri , Saxena Renu 
Published: October 2008

SummaryThrombin activatable fibrinolysis inhibitor [carboxypeptidase B2 (plasma), CPB2] is a basic carboxypeptidase, which inhibits fibrinolysis by cleaving the C‐terminal lysine residues on plasmin‐modified partially degraded fibrin. Plasma CPB2 concentrations have been reported to be under the control of numerous single nucleotide polymorphisms located in the regulatory and coding regions of the gene encoding CPB2 (CPB2). High functional CPB2 levels have been found to be associated with an increased risk for ischemic stroke. The present study investigated CPB2 antigen levels and associated CPB2 polymorphisms in an acute onset non‐cardioembolic stroke population compared with an age‐ and sex‐matched healthy control population. This is, to the best of our knowledge, the first such study done in an Asian Indian population. CPB2 antigen levels were significantly associated with the disease phenotype (P < 0·001) and with CPB2 polymorphisms (P < 0·001). The haplotypes generated on analysis of the genotypic data accounted for 21% of the natural variation in the CPB2 antigenic levels. However none of the haplotype combinations generated showed any association with disease phenotype and therefore could not explain for the difference in CPB2 antigen levels between cases and controls.
Treosulfan‐based conditioning regimen for allogeneic haematopoietic stem cell transplantation in patients with thalassaemia major
Authors: Bernardo Maria Ester , Zecca Marco , Piras Eugenia , Vacca Adriana , Giorgiani Giovanna , Cugno Chiara , Caocci Giovanni , Comoli Patrizia , Mastronuzzi Angela , Merli Pietro , La Nasa Giorgio , Locatelli Franco 
Published: October 2008

SummaryThe safety and efficacy of a preparation with treosulfan/thiotepa/fludarabine were explored in 20 thalassaemia patients given allogeneic marrow transplantation. Seventeen patients were transplanted from unrelated donors after receiving anti‐thymocyte globulin. The regimen was well tolerated. Two patients experienced secondary graft failure; one died of acute graft‐versus‐host disease. Cumulative incidence (95% confidence interval, CI) of transplantation‐related mortality and graft failure was 5% (95% CI, 0–34%) and 11% (95% CI, 3–43%), respectively. Two‐year probability of survival and thalassaemia‐free survival was 95% (95% CI, 85–100%) and 85% (95% CI, 66–100%), respectively. This regimen might find elective application in patients at high risk of developing life‐threatening complications.
Understanding haemophilic arthropathy: an exploration of current open issues
Authors: Jansen Nathalie W. D. , Roosendaal Goris , Lafeber Floris P. J. G. 
Published: October 2008

SummaryHaemophilic arthropathy is joint damage evolving from recurrent joint bleeds that occur in patients suffering from the clotting disorder haemophilia. Insight into the pathogenetic mechanism of this blood‐induced arthropathy yields possible treatment targets and modalities useful to reduce this invalidating co‐morbidity of haemophilia. Joint bleeding leads to initially independent adverse changes in both the synovial tissue and the articular cartilage. These subsequently influence each other: the synovial inflammatory changes enhancing cartilage damage and vice versa. Consequently, effective treatment strategies will have to affect both pathways.
Salinity tolerance of Great Lakes invaders
Authors: ELLIS SANDRA , MACISAAC HUGH J. 
Published: October 2008

Summary1. The Laurentian Great Lakes are among the most invaded freshwater ecosystems in the world. Historically, the major vector for the introduction of non‐indigenous species (NIS) has been the release of contaminated ballast water via transoceanic ships. Despite regulations implemented in 1993, requiring vessels carrying fresh ballast water to exchange this water with saline ocean water, new reports of invasions have continued.2. NIS often have a wide environmental tolerance allowing them to adapt to and invade a variety of habitats. It has been hypothesized that NIS with broad salinity tolerance may be able to survive ballast water exchange (BWE) and continue to pose an invasion risk to the Great Lakes.3. We tested the short‐term salinity tolerance of eight recent invaders to the Great Lakes, specifically three cladocera (Bosmina coregoni, Bythotrephes longimanus, Cercopagis pengoi), two molluscs (Dreissena polymorpha, Dreissena rostriformis bugensis), and one species each of the families Gammaridae, Mysidae and Gobidae (Echinogammarus ischnus, Hemimysis anomala, Neogobius melanostomus) to determine if they could have survived salinities associated with BWE.4. Overall, short‐term exposure to highly saline water dramatically reduced survival of all species. Two different methods of BWE tested, simultaneous and sequential, were equally effective in reducing survival. Species that survived the longest in highly saline water either possess behavioural characteristics that reduce exposure to adverse environments (valve closure; both Dreissena species) or are reported to have some degree of salinity tolerance in their native region (Echinogammarus). Given that exposure in our trials lasted a maximum of 48 h, and that species in ballast tanks would typically be exposed to saline water for c. 5 days, it appears that BWE is an effective method to reduce the survival of these NIS. These results provide impetus for tightening policy and monitoring of BWE, in particular for ships entering the Great Lakes from freshwater ports.