bluuebunny/crossref_metadata_2025_split huggingface dataset:
X‐inactivation is stably maintained in mouse embryos deficient for histone methyl transferase G9a
Authors: Ohhata Tatsuya , Tachibana Makoto , Tada Masako , Tada Takashi , Sasaki Hiroyuki , Shinkai Yoichi , Sado Takashi 
Published: October 2004

AbstractSummary: One of the two X chromosomes becomes inactivated during early development of female mammals. Recent studies demonstrate that the inactive X chromosome is rich in histone H3 methylated at Lys‐9 and Lys‐27, suggesting an important role for these modifications in X‐inactivation. It has been shown that in the mouse Eed is required for maintenance of X‐inactivation in the extraembryonic lineages. Interestingly, Eed associates with Ezh2 to form a complex possessing histone methyltransferase activity predominantly for H3 Lys‐27. We previously showed that G9a is one of the histone methyltransferases specific for H3 Lys‐9 and is essential for embryonic development. Here we examined X‐inactivation in mouse embryos deficient for G9a. Expression of Xist, which is crucial for the initiation of X‐inactivation, was properly regulated and the inactivated X chromosome was stably maintained even in the absence of G9a. These results demonstrate that G9a is not essential for X‐inactivation. genesis 40:151–156, 2004. © 2004 Wiley‐Liss, Inc.
Sox10‐rtTA mouse line for tetracycline‐inducible expression of transgenes in neural crest cells and oligodendrocytes
Authors: Ludwig Andreas , Schlierf Beate , Schardt Anke , Nave Klaus‐Armin , Wegner Michael 
Published: October 2004

AbstractUsing gene targeting, we inserted a high‐affinity variant of the reverse tetracycline controlled transactivator (rtTA) into the genomic Sox10 locus. This rtTA transgene faithfully recapitulated Sox10 expression in the emerging neural crest, several of its derivatives, and in oligodendrocytes. It was furthermore able to induce expression of a tetracycline inducible transgenic reporter gene in a doxycycline‐dependent manner. Induction was fast, with substantial reporter gene expression visible 6 h after the onset of doxycycline treatment. Shut‐off, in contrast, exhibited delayed kinetics, which probably correlated with doxycycline clearance rates. This mouse provides a useful tool for generating tetracycline‐controlled gene expression in neural crest and oligodendrocytes. genesis 40:171–175, 2004. © 2004 Wiley‐Liss, Inc.
NG2 proteoglycan‐expressing cells of the adult rat brain: Possible involvement in the formation of glial scar astrocytes following stab wound
Authors: Alonso G. 
Published: October 2004

AbstractStab wound lesion to the adult central nervous system induces strong proliferative response that is followed by the formation of a dense astroglial scar. In order to determine the origin of those astrocytes composing the glial scar, the cell proliferation marker bromodeoxyuridine (BrdU) was administered to lesioned rats that were fixed 3 h or 6 days later. At 3 h after the BrdU administration, labeled nuclei were frequently associated with either NG2+ cells or microglia/macrophages, but rarely with astrocytes expressing glial fibrillary acidic protein (GFAP). Six days later, by contrast, numerous BrdU‐labeled nuclei were associated with astrocytes located along the lesion borders. After the injection of a viral vector of the green fluorescent protein (GFP) into the lesional cavity, GFP was preferentially detected within NG2‐ or GFAP‐labeled cells when lesioned animals were fixed 1 or 6 days after the injections, respectively. The combined detection of glial markers within cells present in the lesioned area indicated that, although they rarely express GFAP, the marker of mature astrocytes, NG2+ cells located along the lesion borders frequently express nestin and vimentin, i.e., two markers of immature astrocytes. Lastly, chronic treatment of lesioned rats with dexamethasone was found to inhibit the proliferation of NG2+ cells present within the lesioned area and to subsequently alter the formation of a dense astroglial scar. Taken together, these data strongly suggest that following a surgical lesion, at least a portion of the astrocytes that constitute the glial scar are issued from resident NG2+ cells. © 2004 Wiley‐Liss, Inc.
Thrombotic thrombocytopenic purpura: 24 years of experience at the American University of Beirut Medical Center
Authors: Shamseddine Ali , Saliba Teddy , Aoun Elie , Chahal Aref , El‐Saghir Nagi , Salem Ziad , Bazarbachi Ali , Khalil Mazen , Taher Ali 
Published: October 2004

AbstractThrombotic thrombocytopenic purpura (TTP) is a hematological syndrome defined by the presence of thrombocytopenia and microangiopathic hemolytic anemia without a clinically apparent etiology. Patients may also suffer from fever in addition to neurological and renal impairment. Treatment should be initiated as soon as possible, otherwise this rare disease can be fatal. The main treatment options include therapeutic plasma exchange, fresh frozen plasma infusion, and adjuvant agents such as steroids and antiplatelet drugs. A search of patient records was carried out at the American University of Beirut Medical Center looking for patients who developed TTP over a 24‐year period extending from 1980 to 2003. Relevant information was collected and analyzed. A total of 47 records were found. All presented with anemia and thrombocytopenia, 83% had neurological symptoms, 61.7% had fever and 34% had renal impairment. All patients were treated with a multimodality regimen including therapeutic plasma exchange, FFP infusion, steroids, antiplatelet agents, vincristine and others. 38 (81%) cases achieved complete remission. Out of these, 12 (31.6%) relapsed and responded to treatment. Patients who did not receive plasma exchange were more likely to relapse (P = 0.032). A second relapse was observed in 6 cases. The overall mortality rate from TTP over 24 years was 21.3%. TTP remains a fatal disease. A high index of suspicion should, therefore, always be present. Treatment options should be further developed and patients should directly be referred to tertiary care centers. J. Clin. Apheresis 19:119–124, 2004. © 2004 Wiley‐Liss, Inc.
Comparative study of plateletpheresis using Baxter CS 3000 plus and Haemonetics MCS 3P
Authors: Patel Ashwin P. , Kaur Amarjit , Patel Vinod , Patel Narendra , Shah Dilip , Kanvinde Sunil , Prajapati Sanjay , Patel Hiral , Rathod Dinesh , Adesara Rashmin , Rani Shubha 
Published: October 2004

AbstractPlatelet concentrates made from cell separators are used more frequently due to less donor exposure and leucodepletion. This retrospective study was done to compare plateletpheresis done on two cell separators: Baxter CS 3000 plus and Haemonetics MCS 3p. Plateletpheresis procedures, done from January 1997 to April 2002, were included in the study. One hundred and seven procedures were done on Haemonetics MCS 3p using SDP protocol, 49 procedures were done on Haemonetics MCS 3p using PLP protocol, and 107 were done on Baxter CS 3000 plus. Pre‐procedure donor's platelet count and haemoglobin were comparable in all the groups. Platelet yield was comparable in PLP (6.44 × 1011 platelets) and SDP (5.27 × 1011) protocols, but significantly less in Baxter (4.05 × 1011 platelets, P < 0.001 for PLP and P < 0.05 for SDP). Efficiency of platelet removal was statistically significantly different in all the groups (P < 0.0001), however it was more in PLP (PLP‐55.02%, SDP‐47.38%, Baxter 38.98%). A significant number of products (19.51%) of Baxter failed to comply platelet count of product ≤2,435 × 109/l compared to 5.13% in PLP and 1.23% in SDP group; 36.96% of units from PLP and 28% from SDP qualified for split products compared to 1.18% of Baxter. PLP protocol of Haemonetics MCS 3p gives better platelet yield compared to Baxter CS 3000 plus and SDP protocol of Haemonetics MCS 3p. J. Clin. Apheresis 19:137–141, 2004. © 2004 Wiley‐Liss, Inc.
Donor tolerance and results of stimulation with G‐CSF alone or in combination with dexamethasone for the collection of granulocytes
Authors: Moog Rainer 
Published: October 2004

AbstractStimulation of healthy granulocyte donors allows the collection of therapeutic doses of granulocytes. The stimulation with G‐CSF alone was compared with G‐CSF plus dexamethasone. Blood samples were drawn at baseline, at leukapheresis, and at follow‐up visit. Donors answered a questionnaire to evaluate side effects of the stimulation regimen. The combination of G‐CSF and dexamethasone resulted in higher WBC count than G‐CSF alone (39.4 ± 7.8 vs. 34.8 ± 8.3/nl). Glucose (136 ± 45 mg/dl) and lactate dehydrogenase (195 ± 38) increased significantly after stimulation with G‐CSF plus dexamethasone but returned to baseline levels at the follow‐up visit. Generally, stimulation was well tolerated by the donors. A higher rate of mild bone pain and headache was experienced in donors stimulated with G‐CSF plus dexamethasone than in donors receiving G‐CSF alone. Fatigue and myalgia were reported at similar rates in both groups. A high proportion of the donors stated that they would accept a further stimulation and granulocyte donation. At the follow‐up visit, blood counts and chemistry had returned to normal values. J. Clin. Apheresis 19:115–118, 2004. © 2004 Wiley‐Liss, Inc.
Collection of a mobilized peripheral blood apheresis product from a patient with mucopolysaccharidosis type VII and subsequent CD34+ cell isolation
Authors: Hofling A. Alex , Sands Mark S. , Lublin Douglas M. , Bauer Gerhard , Devine Steven 
Published: October 2004

AbstractThe effectiveness of bone marrow transplantation for lysosomal storage diseases like mucopolysaccharidosis type VII (MPSVII) suggests that a gene therapy strategy targeting autologous hematopoietic progenitor cells could be successful. Given the severe systemic manifestations of MPSVII including storage disease in the bone and bone marrow, it was unclear whether sufficient numbers of hematopoietic progenitor cells (CD34+) could be mobilized into the peripheral circulation and subsequently purified from these patients. As reported here, G‐CSF mobilization and apheresis were successful, providing a product of 4 × 1010 nucleated cells containing 0.3% CD34+ progenitors. CD34+ cells were magnetically separated from the product to a final purity of 85% with a 64% yield. These results indicate that hematopoietic progenitors can safely be gathered from an MPSVII patient in numbers sufficient for the trial of clinical gene therapy applications. J. Clin. Apheresis 19:151–153, 2004. © 2004 Wiley‐Liss, Inc.
Assessment of complement activation during membrane‐based plasmapheresis procedures
Authors: Burnouf Thierry , Eber Michel , Kientz Daniel , Cazenave Jean‐Pierre , Burkhardt Thomas 
Published: October 2004

AbstractPrevious studies have suggested that plasmapheresis procedures using a separation membrane may activate the complement system and release anaphylatoxins. This study determines the content in C3a/C3ades Arg and C5a/C5ades Arg in plasma donations obtained by the new Haemonetics® Filter Core (FC) procedure and compares it to Baxter Autopheresis C® (Auto‐C). FC performs sequential blood centrifugation and plasma filtration on a microporous polyethersulfone membrane, while Auto‐C removes blood cells by simultaneous gravitation and filtration on a rotating nylon membrane. One group of 34 donors donated on FC and two groups of 30 and 10 donors on Auto‐C. Plasma aliquots were taken from the plasma units within 30 min of the end of the collection procedures, frozen at <−30°C and assessed for C3a and C5a at various time points of storage. Mean C3a/C3ades Arg in FC plasma (N = 34) was 1,151 (range: 526–2,991), 1,092 (range: 349–3498), and 507 (range: 307–815) ng/ml at time of collection and after 6 and 12 months of storage, respectively. Respective C5a/C5ades Arg was 26.6 (range 4.9–74), 18.9 (9.5–42.6), and 30.9 (range: 10.7–62.3) ng/ml. Mean C3a/C3ades Arg was higher in Auto‐C (P < 0.001): 4,724 ng/ml (N = 10; range: 2,400–7 ,360) and >4,149 ng/ml (N = 30; 2,408–>6,430) after 3 and 18 months storage, respectively. Mean C5a/C5ades Arg was 32.1 ng/ml (N = 30; range: 10.6–57.2) after 18 months of storage. Complement activation in FC plasmas appears limited compared to Auto‐C, suggesting better biocompatibility of this collection device and/or a favourable impact of the sequential cell centrifugation/filtration technology used. Further studies are needed to explain differences in complement activation between apheresis procedures and to assess clinical impacts, if any. J. Clin. Apheresis, 19:142–147, 2004. © 2004 Wiley‐Liss, Inc.
Plasmapheresis in acute disseminated encephalomyelitis
Authors: Lin Chin‐Hsien , Jeng Jiann‐Shing , Yip Ping‐Keung 
Published: October 2004

AbstractHigh‐dose corticosteroid administration has been regarded as the main therapy for acute disseminated encephalomyelitis (ADEM). However, some patients with ADEM do not respond well to this treatment. We successfully used plasmapheresis to treat 2 patients who had ADEM. We also compared our patients' symptoms and clinical outcomes to those from previous reports. Plasmapheresis may be indicated not only for severe cases that fail to improve after high‐dose corticosteroid treatment but also for first‐line treatment. Additional large, controlled, double‐blinded trials are needed to clarify the role of plasmapheresis in ADEM. J Clin. Apheresis 19:154–159, 2004. © 2004 Wiley‐Liss, Inc.
Initiation of DNA replication at a nuclear matrix‐attached chromatin fraction
Authors: Radichev Ilian , Parashkevova Anastassia , Anachkova Boyka 
Published: October 2004

AbstractIt is still unclear what nuclear components support initiation of DNA replication. To address this issue, we developed a cell‐free replication system in which the nuclear matrix along with the residual matrix‐attached chromatin was used as a substrate for DNA replication. We found out that initiation occurred at late G1 residual chromatin but not at early G1 chromatin and depended on cytosolic and nuclear factors present in S phase cells but not in G1 cells. Initiation of DNA replication occurred at discrete replication foci in a pattern typical for early S phase. To prove that the observed initiation takes place at legitimate DNA replication origins, the in vitro synthesized nascent DNA strands were isolated and analyzed. It was shown that they were enriched in sequences from the core origin region of the early firing, dihydrofolate reductase origin of replication ori‐β and not in distal to the origin sequences. A conclusion is drawn that initiation of DNA replication occurs at discrete sub‐chromosomal structures attached to the nuclear matrix. © 2004 Wiley‐Liss, Inc.
JAK and STAT proteins are expressed and activated by IFN‐γ in rat pancreatic acinar cells
Authors: Gallmeier E. , Schäfer C. , Moubarak P. , Tietz A. , Plössl I. , Huss R. , Göke B. , Wagner A.C.C. 
Published: October 2004

AbstractThe development of acute pancreatitis (AP) is triggered by acinar events, but the subsequent extra‐acinar events, particularly a distinct immune response, appear to determine its severity. Cytokines modulate this immune response and are derived not only from immunocytes but also from pancreatic acinar cells. We studied whether pancreatic acinar cells were also capable of responding to cytokines. The JAK/STAT‐pathway represents the main effector for many cytokines. Therefore, expression and regulation of JAK and STAT proteins were investigated in rat pancreatic acinar cells. Western blotting showed expression of JAK1, JAK2, Tyk2, and STAT1, STAT2, STAT3, STAT5, STAT6. In addition, STAT1 was reversibly tyrosine‐phosphorylated upon the procedure of acinar cell isolation. In contrast, STAT3‐phosphorylation occurred spontaneously after pancreas removal and was not reversible within 8 h. STAT1 phosphorylation was also observed upon treatment with IFN‐γ but not upon EGF, TNF‐α or IL‐6, and inhibited by the JAK2‐inihibitor AG‐490. Immunohistochemistry revealed cytoplasmic expression of unphosphorylated STAT1 in untreated acinar cells and nuclear translocation of phosphorylated STAT1 following IFN‐γ‐treatment. Interestingly, although CCK leads to the activation of multiple stress pathways in pancreatic acinar cells, we found no influence of CCK on phosphorylation of STAT1, STAT3, or STAT5 in the pancreas. In conclusion, our data provide further evidence that pancreatic acinar cells are able to interact with immune cells. Besides stimulating immune cells via cytokine secretion, acinar cells are in turn capable of responding to IFN‐γ via JAK2 and STAT1 which may have an impact on the development of AP. © 2004 Wiley‐Liss, Inc.
Glioblastoma cells deficient in DNA‐dependent protein kinase are resistant to cell death
Authors: Chen George G. , Sin Fanny L.F. , Leung Billy C.S. , Ng Ho K. , Poon Wai S. 
Published: October 2004

AbstractDNA‐dependent protein kinase (DNA‐PK), a nuclear serine/threonine kinase, is responsible for the DNA double‐strand break repair. Cells lacking or with dysfunctional DNA‐PK are often associated with mis‐repair, chromosome aberrations, and complex exchanges, all of which are known to contribute to the development of human cancers including glioblastoma. Two human glioblastoma cell lines were used in the experiment, M059J cells lacking the catalytic subunit of DNA‐PK, and their isogenic but DNA‐PK proficient counterpart, M059K. We found that M059K cells were much more sensitive to staurosporine (STS) treatment than M059J cells, as demonstrated by MTT assay, TUNEL detection, and annexin‐V and propidium iodide (PI) staining. A possible mechanism responsible for the different sensitivity in these two cell lines was explored by the examination of Bcl‐2, Bax, Bak, and Fas. The cell death stimulus increased anti‐apoptotic Bcl‐2 and decreased pro‐apoptotic Bcl‐2 members (Bak and Bax) and Fas in glioblastoma cells deficient in DNA‐PK. Activation of DNA‐PK is known to promote cell death of human tumor cells via modulation of p53, which can down‐regulate the anti‐apoptotic Bcl‐2 member proteins, induce pro‐apoptotic Bcl‐2 family members and promote a Bax–Bak interaction. Our experiment also demonstrated that the mode of glioblastoma cell death induced by STS consisted of both apoptosis and necrosis and the percentage of cell death in both modes was similar in glioblastoma cell lines either lacking DNA‐PK or containing intact DNA‐PK. Taken together, our findings suggest that DNA‐PK has a positive role in the regulation of apoptosis in human glioblastomas. The aberrant expression of Bcl‐2 family members and Fas was, at least in part, responsible for decreased sensitivity of DNA‐PK deficient glioblastoma cells to cell death stimuli. © 2004 Wiley‐Liss, Inc.
Specific AHNAK expression in brain endothelial cells with barrier properties
Authors: Gentil Benoît J. , Benaud Christelle , Delphin Christian , Remy Chantal , Berezowski Vincent , Cecchelli Roméo , Feraud Olivier , Vittet Daniel , Baudier Jacques 
Published: October 2004

AbstractThe blood–brain barrier (BBB) is essential for maintaining brain homeostasis and low permeability. Because disruption of the BBB may contribute to many brain disorders, they are of considerable interests in the identification of the molecular mechanisms of BBB development and integrity. We here report that the giant protein AHNAK is expressed at the plasma membrane of endothelial cells (ECs) forming specific blood–tissue barriers, but is absent from the endothelium of capillaries characterized by extensive molecular exchanges between blood and extracellular fluid. In the brain, AHNAK is widely distributed in ECs with BBB properties, where it co‐localizes with the tight junction protein ZO‐1. AHNAK is absent from the permeable brain ECs of the choroid plexus and is down‐regulated in permeable angiogenic ECs of brain tumors. In the choroid plexus, AHNAK accumulates at the tight junctions of the choroid epithelial cells that form the blood–cerebrospinal fluid (CSF) barrier. In EC cultures, the regulation of AHNAK expression and its localization corresponds to general criteria of a protein involved in barrier organization. AHNAK is up‐regulated by angiopoietin‐1 (Ang‐1), a morphogenic factor that regulates brain EC permeability. In bovine cerebral ECs co‐cultured with glial cells, AHNAK relocates from the cytosol to the plasma membrane when endothelial cells acquire BBB properties. Our results identify AHNAK as a protein marker of endothelial cells with barrier properties. © 2004 Wiley‐Liss, Inc.
Reorganization of ZO‐1 by sodium‐dependent glucose transporter activation after heat stress in LLC‐PK1 cells
Authors: Ikari Akira , Nakano Mika , Suketa Yasunobu , Harada Hitoshi , Takagi Kuniaki 
Published: October 2004

AbstractHeat stress (HS) induces activation of high‐affinity sodium‐dependent glucose transporter (SGLT1) in porcine renal LLC‐PK1 cells. In this study, we investigated the roles of SGLT1 activation in reorganization of zonula occludens‐1 (ZO‐1), a cytosolic tight junction (TJ) protein, after HS. HS (42°C, 3 h) caused decrease in transepithelial electrical resistance (TER). Subsequent incubation at 37°C for 12 h increased TER above pre‐HS level. The treatment of phloridzin, a potent SGLT1 inhibitor, or the replacement of glucose with a nonmetabolizable glucose analog blocked the recovery of TER and increased the transepithelial flux of FITC‐dextran (4,000 Da). Immunofluorescent staining of ZO‐1 showed that HS diffused ZO‐1 from cell contact to cytosolic sites. Furthermore, the fraction of ZO‐1 was distributed from the Triton X‐100 insoluble to the Triton X‐100 soluble pool. After incubation at 37°C for 12 h, cell contact and ZO‐1 extractability with Triton X‐100 returned to pre‐HS conditions, but the recovery was completely prevented by phloridzin. Tyrosine kinases activity was increased by HS that was inhibited by phloridzin. Genistein and CGP77675, tyrosine kinases inhibitors, blocked the recovery of TER and increased the transepithelial flux of FITC‐dextran. Furthermore, these inhibitors prevented the recovery of cell contact and ZO‐1 extractability with Triton X‐100 as same as phloridzin. These findings suggested that the activation of SGLT1 reorganized ZO‐1 mediated by elevation of tyrosine kinases activity after heat injury. © 2004 Wiley‐Liss, Inc.