bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytotoxic activity of the maytansinoid immunoconjugate B-B4–DM1 against CD138+ multiple myeloma cells
Authors: Tassone Pierfrancesco , Goldmacher Victor S. , Neri Paola , Gozzini Antonella , Shammas Masood A. , Whiteman Kathleen R. , Hylander-Gans Linda L. , Carrasco Daniel R. , Hideshima Teru , Shringarpure Reshma , Shi Jialan , Allam Charles K. , Wijdenes John , Venuta Salvatore , Munshi Nikhil C. , Anderson Kenneth C.
Published: August 2004

We tested the in vitro and in vivo antitumor activity of the maytansinoid DM1 (N2′-deacetyl-N2′-(3-mercapto-1-oxopropyl)-maytansine), a potent antimicrotubule agent, covalently linked to the murine monoclonal antibody (mAb) B-B4 targeting syndecan-1 (CD138). We evaluated the in vitro activity of B-B4–DM1 against a panel of CD138+ and CD138- cell lines, as well as CD138+ patient multiple myeloma (MM) cells. Treatment with B-B4–DM1 selectively decreased growth and survival of MM cell lines, patient MM cells, and MM cells adherent to bone marrow stromal cells. We further examined the activity of B-B4–DM1 in 3 human MM models in mice: (1) severe combined immunodeficient (SCID) mice bearing subcutaneous xenografts; (2) SCID mice bearing green fluorescent protein–positive (GFP+) xenografts; and (3) SCID mice implanted with human fetal bone (SCID-hu) and subsequently injected with patient MM cells. Tumor regression and inhibition of tumor growth, improvement in overall survival, and reduction in levels of circulating human paraprotein were observed in mice treated with B-B4–DM1. Although immunohistochemical analysis demonstrates restricted CD138 expression in human tissues, the lack of B-B4 reactivity with mouse tissues precludes evaluation of its toxicity in these models. In conclusion, B-B4–DM1 is a potent anti-MM agent that kills cells in an antigen-dependent manner in vitro and mediates in vivo antitumor activity at doses that are well tolerated, providing the rationale for clinical trials of this immunoconjugate in MM.
Successful correction of the human β-thalassemia major phenotype using a lentiviral vector
Authors: Puthenveetil Geetha , Scholes Jessica , Carbonell Denysha , Qureshi Naveen , Xia Ping , Zeng Licheng , Li Shulian , Yu Ying , Hiti Alan L , Yee Jiing-Kuan , Malik Punam
Published: August 2004

β-thalassemias are the most common single gene disorders and are potentially amenable to gene therapy. However, retroviral vectors carrying the human β-globin cassette have been notoriously unstable. Recently, considerable progress has been made using lentiviral vectors, which stably transmit the β-globin expression cassette. Thus far, mouse studies have shown correction of the β-thalassemia intermedia phenotype and a partial, variable correction of β-thalassemia major phenotype. We tested a lentiviral vector carrying the human β-globin expression cassette flanked by a chromatin insulator in transfusion-dependent human thalassemia major, where it would be ultimately relevant. We demonstrated that the vector expressed normal amounts of human β-globin in erythroid cells produced in in vitro cultures for unilineage erythroid differentiation. There was restoration of effective erythropoiesis and reversal of the abnormally elevated apoptosis that characterizes β-thalassemia. The gene-corrected human β-thalassemia progenitor cells were transplanted into immune-deficient mice, where they underwent normal erythroid differentiation, expressed normal levels of human β-globin, and displayed normal effective erythropoiesis 3 to 4 months after xenotransplantation. Variability of β-globin expression in erythroid colonies derived in vitro or from xenograft bone marrow was similar to that seen in normal controls. Our results show genetic modification of primitive progenitor cells with correction of the human thalassemia major phenotype.
L-Selectinhi but not the L-selectinlo CD4+25+ T-regulatory cells are potent inhibitors of GVHD and BM graft rejection
Authors: Taylor Patricia A. , Panoskaltsis-Mortari Angela , Swedin Jessica M. , Lucas Philip J. , Gress Ronald E. , Levine Bruce L. , June Carl H. , Serody Jonathan S. , Blazar Bruce R.
Published: August 2004

Graft-versus-host disease (GVHD) is a major cause of morbidity and mortality after bone marrow transplantation (BMT). CD4+CD25+ immune regulatory T cells (Tregs), long recognized for their critical role in induction and maintenance of self-tolerance and prevention of autoimmunity, are also important in the regulation of immune responses in allogeneic bone marrow (BM) and solid organ transplantation. Published data indicate that ex vivo activated and expanded donor Tregs result in significant inhibition of lethal GVHD. This study provides a direct comparison of LSelhi and LSello Tregs for GVHD inhibition and for the promotion of allogeneic BM engraftment. Imaging of green fluorescent protein–positive effectors in GVHD control mice and LSelhi and LSello Treg-treated mice vividly illustrate the multisystemic nature of GVHD and the profound inhibition of GVHD by LSelhi Tregs. Data indicate that LSelhi Tregs interfere with the activation and expansion of GVHD effector T cells in secondary lymphoid organs early after BMT. Either donor- or host-type LSelhi, but not LSello, Tregs potently increased donor BM engraftment in sublethally irradiated mice, an event occurring independently of transforming growth factor β signaling of host T cells. These data indicate that Treg cellular therapy warrants clinical consideration for the inhibition of GVHD and the promotion of alloengraftment.
Maternal microchimerism in the livers of patients with Biliary atresia
Authors: Suskind David L , Rosenthal Philip , Heyman Melvin B , Kong Denice , Magrane Greg , Baxter-Lowe Lee-Ann , Muench Marcus O
Published: August 2004

Background

Biliary atresia (BA) is a neonatal cholestatic disease of unknown etiology. It is the leading cause of liver transplantation in children. Many similarities exist between BA and graft versus host disease suggesting engraftment of maternal cells during gestation could result in immune responses that lead to BA. The aim of this study was to determine the presence and extent of maternal microchimerism (MM) in the livers of infants with BA.

Methods

Using fluorescent in situ hybridization (FISH), 11 male BA & 4 male neonatal hepatitis (NH) livers, which served as controls, were analyzed for X and Y-chromosomes. To further investigate MM in BA, 3 patients with BA, and their mothers, were HLA typed. Using immunohistochemical stains, the BA livers were examined for MM. Four additional BA livers underwent analysis by polymerase chain reaction (PCR) for evidence of MM.

Results

By FISH, 8 BA and 2 NH livers were interpretable. Seven of eight BA specimens showed evidence of MM. The number of maternal cells ranged from 2–4 maternal cells per biopsy slide. Neither NH specimen showed evidence of MM. In addition, immunohistochemical stains confirmed evidence of MM. Using PCR, a range of 1–142 copies of maternal DNA per 25,000 copies of patients DNA was found.

Conclusions

Maternal microchimerism is present in the livers of patients with BA and may contribute to the pathogenesis of BA.
The usefulness of the Korean version of modified Mini-Mental State Examination (K-mMMSE) for dementia screening in community dwelling elderly people
Authors: Jeong Seul-Ki , Cho Ki-Hyun , Kim Jae-Min
Published: August 2004

Background

We assessed whether the Korean version of modified Mini-Mental State Examination (K-mMMSE) has improved performance as a screening test for cognitive impairment or dementia in a general population compared with the Korean Mini-Mental State Examination (K-MMSE).

Methods

Screening interviews were conducted with people aged 65 and over in Noam-dong, Namwon-city, Jeonbuk province. There were 522 community participants, of whom 235 underwent clinical and neuropsychological examination for diagnosis of dementia and Cognitive Impairment No Dementia (CIND). Sensitivity, specificity and areas under the receiver operating characteristic (ROC) curves for the K-mMMSE and the K-MMSE were the main outcome measures.

Results

Cronbach's alpha for the K-mMMSE was 0.91, compared with 0.84 for the K-MMSE. The areas under the ROC curves in identifying all levels of CIND or dementia were 0.91 for the K-mMMSE and 0.89 for the K-MMSE (P < 0.05). For the K-mMMSE, the optimal cut-off score for a diagnosis of CIND was 69/70, which had a sensitivity of 0.86 and a specificity of 0.79, while, for a diagnosis of dementia, the optimal cut-off score of 59/60 had a sensitivity of 0.91 and a specificity of 0.78. The K-mMMSE also had a high test-retest reliability (r = 0.89).

Conclusion

Our findings indicate that the K-mMMSE is more reliable and valid than the K-MMSE as a cognitive screen in a population based study of dementia. Considering the test characteristics, the K-MMSE and modified version are expected to be optimally used in clinical and epidemiologic fields.
Sad, blue, or depressed days, health behaviors and health-related quality of life, Behavioral Risk Factor Surveillance System, 1995–2000
Authors: Kobau Rosemarie , Safran Marc A , Zack Matthew M , Moriarty David G , Chapman Daniel
Published: August 2004

Background

Mood disorders are a major public health problem in the United States as well as globally. Less information exists however, about the health burden resulting from subsyndromal levels of depressive symptomatology, such as feeling sad, blue or depressed, among the general U.S. population.

Methods

As part of an optional Quality of Life survey module added to the U.S. Behavioral Risk Factor Surveillance System, between 1995–2000 a total of 166,564 BRFSS respondents answered the question, "During the past 30 days, for about how many days have you felt sad, blue, or depressed?" Means and 95% confidence intervals for sad, blue, depressed days (SBDD) and other health-related quality of life (HRQOL) measures were calculated using SUDAAN to account for the BRFSS's complex sample survey design.

Results

Respondents reported a mean of 3.0 (95% CI = 2.9–3.1) SBDD in the previous 30 days. Women (M = 3.5, 95% CI = 3.4–3.6) reported a higher number of SBDD than did men (M = 2.4, 95% CI = 2.2–2.5). Young adults aged 18–24 years reported the highest number of SBDD, whereas older adults aged 60–84 reported the fewest number. The gap in mean SBDD between men and women decreased with increasing age. SBDD was associated with an increased prevalence of behaviors risky to health, extremes of body mass index, less access to health care, and worse self-rated health status. Mean SBDD increased with progressively higher levels of physically unhealthy days, mentally unhealthy days, unhealthy days, activity limitation days, anxiety days, pain days, and sleepless days.

Conclusion

Use of this measure of sad, blue or depressed days along with other valid mental health measures and community indicators can help to assess the burden of mental distress among the U.S. population, identify subgroups with unmet mental health needs, inform the development of targeted interventions, and monitor changes in population levels of mental distress over time.
Public – private 'partnerships' in health – a global call to action
Authors: Nishtar Sania
Published: August 2004

The need for public-private partnerships arose against the backdrop of inadequacies on the part of the public sector to provide public good on their own, in an efficient and effective manner, owing to lack of resources and management issues. These considerations led to the evolution of a range of interface arrangements that brought together organizations with the mandate to offer public good on one hand, and those that could facilitate this goal though the provision of resources, technical expertise or outreach, on the other. The former category includes of governments and intergovernmental agencies and the latter, the non-profit and for-profit private sector. Though such partnerships create a powerful mechanism for addressing difficult problems by leveraging on the strengths of different partners, they also package complex ethical and process-related challenges. The complex transnational nature of some of these partnership arrangements necessitates that they be guided by a set of global principles and norms. Participation of international agencies warrants that they be set within a comprehensive policy and operational framework within the organizational mandate and involvement of countries requires legislative authorization, within the framework of which, procedural and process related guidelines need to be developed. This paper outlines key ethical and procedural issues inherent to different types of public-private arrangements and issues a Global Call to Action.
Markers of macrophage differentiation in experimental silicosis
Authors: Misson Pierre , van den Brûle Sybille , Barbarin Virginie , Lison Dominique , Huaux François
Published: August 2004

Macrophages are characterized by a marked phenotypic heterogeneity depending on their microenvironmental stimulation. Beside classical activation (M1), it has been shown that macrophages could follow a different activation pathway after stimulation with interleukin (IL)-4 or IL-13 (M2). Recently, it has been postulated that those “alternatively activated” macrophages may be critical in the control of fibrogenesis. In an experimental model of silicosis, where pulmonary macrophages play a central role, we addressed the question of whether lung fibrosis development would be associated with alternative macrophage activation. As available markers for alternative macrophage activation, type-1 arginase (Arg-1), Fizz1, Ym1/2, and mannose receptor expression were evaluated at the mRNA and/or protein levels at different stages of the disease. Nitric oxide synthase-2 (NOS-2) expression was also examined to investigate the classical counterpart. We found that the expression of Arg-1, Fizz1, and NOS-2 in adherent bronchoalveolar lavage cells was highly up-regulated 3 days after silica administration but returned to control levels during the fibrotic stage of the disease (60 days). By comparing the early response to silica in C57BL/6 and BALB/c mice, we observed that the amplitude of Arg-1 mRNA up-regulation was not associated with the severity of lung fibrosis. Using a model of manganese dioxide particles (resolutive alveolitis), we showed that this early Arg-1 mRNA was not specific to a fibrogenic lung response. Our data indicate that the modifications of M1/M2 marker expression are limited to the early inflammatory stage of silicosis and that the establishment of a fibrotic process is not necessarily associated with M2 polarization.
Expression and regulation of NFAT (nuclear factors of activated T cells) in human CD34+ cells: down-regulation upon myeloid differentiation
Authors: Kiani Alexander , Habermann Ivonne , Haase Michael , Feldmann Silvia , Boxberger Sabine , Sanchez-Fernandez Maria A , Thiede Christian , Bornhäuser Martin , Ehninger Gerhard
Published: August 2004

The calcineurin-dependent, cyclosporin A (CsA)-sensitive transcription factor nuclear factor of activated T cells (NFAT) represents a group of proteins, which is well-characterized as a central regulatory element of cytokine expression in activated T cells. In contrast, little is known about the expression or function of NFAT family members in myeloid cells; moreover, it is unclear whether they are expressed by hematopoietic stem/progenitor cells. Here, we show that NFATc2 (NFAT1) is expressed at high levels in CD34+ cells and megakaryocytes but not in cells committed to the neutrophilic, monocytic, or erythroid lineages. Cytokine-induced in vitro differentiation of CD34+ cells into neutrophil granulocytes results in the rapid suppression of NFATc2 RNA and protein. NFATc2 dephosphorylation/rephosphorylation as well as nuclear/cytoplasmic translocation in CD34+ cells follow the same calcineurin-dependent pattern as in T lymphocytes, suggesting that NFATc2 activation in these cells is equally sensitive to inhibition with CsA. Finally, in vitro proliferation, but not differentiation, of CD34+ cells cultured in the presence of fms-like tyrosine kinase 3 ligand (FLT3L), stem cell factor, granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin-3, and G-CSF is profoundly inhibited by treatment with CsA in a dose-dependent manner. These results suggest a novel and unexpected role for members of the NFAT transcription factor family in the hematopoietic system.
Antimicrobial proteins and peptides: anti-infective molecules of mammalian leukocytes
Authors: Levy Ofer
Published: August 2004

Phagocytic leukocytes are a central cellular element of innate-immune defense in mammals. Over the past few decades, substantial progress has been made in defining the means by which phagocytes kill and dispose of microbes. In addition to the generation of toxic oxygen radicals and nitric oxide, leukocytes deploy a broad array of antimicrobial proteins and peptides (APP). The majority of APP includes cationic, granule-associated (poly)peptides with affinity for components of the negatively charged microbial cell wall. Over the past few years, the range of cells expressing APP and the potential roles of these agents have further expanded. Recent advances include the discovery of two novel families of mammalian APP (peptidoglycan recognition proteins and neutrophil gelatinase-associated lipocalin), that the oxygen-dependent and oxygen-independent systems are inextricably linked, that APP can be deployed in the context of novel subcellular organelles, and APP and the Toll-like receptor system interact. From a clinical perspective, congeners of several of the APP have been developed as potential therapeutic agents and have entered clinical trials with some evidence of benefit.
The orphan nuclear receptor SHP is involved in monocytic differentiation, and its expression is increased by c-Jun
Authors: Choi Yoon Ha , Park Min Jung , Kim Kook Whan , Lee Hyung Chul , Choi Young Hyun , Cheong JaeHun
Published: August 2004

Small heterodimer partner (SHP) is an atypical member of nuclear receptor superfamily that lacks a DNA binding domain. Here, we show that SHP expression increases during monocytic differentiaton with exposure HL-60 leukemia cells to a 12-O-tetradecanoylphorbol-13-acetate (TPA) response element, whose treatment induced the SHP promoter activity dependent on c-Jun expression, which is well known to be involved in the commitment step in the TPA-induced differentiation of HL-60 leukemia cells. We also show that overexpression and activation signaling of c-Jun increase the SHP promoter activity, suggesting that the level of SHP expression is normally limiting for c-Jun-dependent monocytic differentiation. Electrophoretic mobility shift assays using oligonucleotides derived from the SHP promoter reveal that c-Jun exhibit TPA-induced DNA binding, providing a mechanism for the transcriptional activation of SHP gene expression. It was also found that overexpression of SHP and c-Jun greatly facilitated monocytic differentiation by TPA and surprisingly, that expression of SHP or c-Jun alone was sufficient to make cells differentiate into functionally mature monocytes, but silencing of SHP and c-Jun by RNA interference diminished the TPA-induced monocytic differentiation. Taken together, these works suggest that c-Jun works to activate the expression of SHP genes associated with the cascade regulation of monocytic differentiation.
Neurogenesis of Rhesus adipose stromal cells
Authors: Kang Soo Kyung , Putnam Lorna A. , Ylostalo Joni , Popescu Ion Razvan , Dufour Jason , Belousov Andrei , Bunnell Bruce A.
Published: August 2004

In this study, we isolated and characterized a population of non-human primate adipose tissue stromal cells (pATSCs) containing multipotent progenitor cells. We show that these pATSCs can differentiate into several mesodermal lineages, as well as neural lineage cells. For neural induction of pATSCs and non-human primate bone marrow stromal cells (pBMSCs), the cells were cultured in Neurobasal (NB) media supplemented with B27, basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and epidermal growth factor (EGF). After 4 days in culture, the pATSCs form compact, spheroid bodies that ultimately become neurospheres (NS). Free-floating neurospheres undergo extensive differentiation when cultured on PDL-laminin. Our data suggest that the neurogenic potential of pATSCs is markedly higher than that of pBMSCs. We have also performed microarray analysis and characterized the gene expression patterns in undifferentiated pATSCs. The direct comparison of gene expression profiles in undifferentiated pATSCs and pATSC-NS, and delineated specific members of important growth factor, signaling, cell adhesion and transcription factors families. Our data indicate that adipose tissue may be an alternative source of stem cells for therapy of central nervous system (CNS) defects.
Sialic acid residues on astrocytes regulate neuritogenesis by controlling the assembly of laminin matrices
Authors: Freire Elisabete , Gomes Flávia C. A. , Jotha-Mattos Tatiana , Neto Vivaldo Moura , Filho Fernando C. Silva , Coelho-Sampaio Tatiana
Published: August 2004

In the developing nervous system migrating neurons and growing axons are guided by diffusible and/or substrate-bound cues, such as extracellular matrix-associated laminin. In a previous work we demonstrated that laminin molecules could self-assemble in two different manners, giving rise to matrices that could favor either neuritogenesis or proliferation of cortical precursor cells. We investigated whether the ability of astrocytes to promote neuritogenesis of co-cultivated neurons was modulated by the assembling mode of the laminin matrix secreted by them. We compared the morphologies and neuritogenic potentials of laminin deposited by in vitro-differentiated astrocytes obtained from embryonic or neonatal rat brain cortices. We showed that, while permissive astrocytes derived from embryonic brain produced a flat laminin matrix that remained associated to the cell surface, astrocytes derived from newborn brain secreted a laminin matrix resembling a fibrillar web that protruded from the cell plane. The average neurite lengths obtained for E16 neurons cultured on each astrocyte layer were 198±22 and 123±13 μm, respectively. Analyses of surface-associated electrostatic potentials revealed that embryonic astrocytes presented a pI of -2.8, while in newborn cells this value was -3.8. Removal of the sialic acid groups on the embryonic monolayer by neuraminidase treatment led to the immediate release of matrix-associated laminin. Interestingly, laminin reassembled 1 hour after neuraminidase removal converted to the features of the newborn matrix. Alternatively, treatment of astrocytes with the cholesterol-solubilizing detergent methyl-β-cyclodextrin also resulted in release of the extracellular laminin. To test the hypothesis that sialic-acid-containing lipids localized at cholesterol-rich membrane domains could affect the process of laminin assembly, we devised a cell-free assay where laminin polymerization was carried out over artificial lipid films. Films of either a mixture of gangliosides or pure ganglioside GT1b induced formation of matrices of morpho-functional features similar to the matrices deposited by embryonic astrocytes. Conversely, films of phosphatidylcholine or ganglioside GM1 led to the formation of bulky laminin aggregates that lacked a defined structure. We propose that the expression of negative lipids on astrocytes can control the extracellular polymerization of laminin and, consequently, the permissivity to neuritogenesis of astrocytes during development.
Simulation of calcium waves in ascidian eggs: insights into the origin of the pacemaker sites and the possible nature of the sperm factor
Authors: Dupont Geneviève , Dumollard Rémi
Published: August 2004

Fertilization triggers repetitive waves of cytosolic Ca2+ in the egg of many species. The mechanism involved in the generation of Ca2+ waves has been studied in much detail in mature ascidian eggs, by raising artificially the level of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] or of its poorly metabolizable analogue, glycero-myo-phosphatidylinositol 4,5-bisphosphate [gPtdIns(4,5)P2]. Here, we use this strategy and the experimental results it provides to develop a realistic theoretical model for repetitive Ca2+ wave generation and propagation in mature eggs. The model takes into account the heterogeneous spatial distribution of the endoplasmic reticulum. Our results corroborate the hypothesis that Ca2+ wave pacemakers are associated with cortical accumulations of endoplasmic reticulum. The model is first tested and validated by the adequate match between its theoretical predictions and the observed effects of localized injections of massive amounts of Ins(1,4,5)P3 analogues. In a second step, we use the model to make some propositions about the possible characteristics of the sperm factor. We find that to account for the spatial characteristics of the first series of Ca2+ waves seen at fertilization in ascidian eggs, it has to be assumed that, if the sperm factor is a phospholipase C, it is Ca2+-sensitive and highly diffusible. Although the actual state of knowledge does not allow us to explain the observed relocalization of the Ca2+ wave pacemaker site, the model corroborates the assumption that PtdIns(4,5)P2, the substrate for phospholipase C is distributed over the entire egg. We also predict that the dose of sperm factor injected into the egg should modulate the temporal characteristics of the first, long-lasting fertilization wave.
Identification of the Drosophila interband-specific protein Z4 as a DNA-binding zinc-finger protein determining chromosomal structure
Authors: Eggert Harald , Gortchakov Andrej , Saumweber Harald
Published: August 2004

The subdivision of polytene chromosomes into bands and interbands suggests a structural chromatin organization that is related to the formation of functional domains of gene expression. We made use of the antibody Z4 to gain insight into this level of chromosomal structure, as the Z4 antibody mirrors this patterning by binding to an antigen that is present in most interbands. The Z4 gene encodes a protein with seven zinc fingers, it is essential for fly development and acts in a dose-dependent manner on the development of several tissues. Z4 mutants have a dose-sensitive effect on wm4 position effect variegation with a haplo-suppressor and triplo-enhancer phenotype, suggesting Z4 to be involved in chromatin compaction. This assumption is further supported by the phenotype of Z4 mutant chromosomes, which show a loss of the band/interband pattern and are subject to an overall decompaction of chromosomal material. By co-immunoprecipitations we identified a novel chromo domain protein, which we named Chriz (Chromo domain protein interacting with Z4) as an interaction partner of Z4. Chriz localizes to interbands in a pattern that is identical to the Z4 pattern. These findings together with the result that Z4 binds directly to DNA in vitro strongly suggest that Z4 in conjunction with Chriz is intimately involved in the higher-order structuring of chromosomes.
Trichostatin A-induced histone acetylation causes decondensation of interphase chromatin
Authors: Tóth Katalin Fejes , Knoch Tobias A. , Wachsmuth Malte , Frank-Stöhr Monika , Stöhr Michael , Bacher Christian P. , Müller Gabriele , Rippe Karsten
Published: August 2004

The effect of trichostatin A (TSA)-induced histone acetylation on the interphase chromatin structure was visualized in vivo with a HeLa cell line stably expressing histone H2A, which was fused to enhanced yellow fluorescent protein. The globally increased histone acetylation caused a reversible decondensation of dense chromatin regions and led to a more homogeneous distribution. These structural changes were quantified by image correlation spectroscopy and by spatially resolved scaling analysis. The image analysis revealed that a chromatin reorganization on a length scale from 200 nm to >1 μm was induced consistent with the opening of condensed chromatin domains containing several Mb of DNA. The observed conformation changes could be assigned to the folding of chromatin during G1 phase by characterizing the effect of TSA on cell cycle progression and developing a protocol that allowed the identification of G1 phase cells on microscope coverslips. An analysis by flow cytometry showed that the addition of TSA led to a significant arrest of cells in S phase and induced apoptosis. The concentration dependence of both processes was studied.
Cpc1, aChlamydomonascentral pair protein with an adenylate kinase domain
Authors: Zhang Hui , Mitchell David R.
Published: August 2004

Mutations at CPC1 disrupt assembly of a central pair microtubule-associated complex and alter flagellar beat frequency in Chlamydomonas. Sequences of wild-type genomic clones that complement cpc1, and of corresponding cDNAs, reveal the gene product to be a 205 kDa protein with two predicted functional domains, a single EF hand motif near the C-terminus and an unusual centrally located adenylate kinase domain. Homologs are expressed in mammals (testis and tracheal cilia) as well as ciliated lower eukaryotes. Western blots confirm that Cpc1 is one of six subunits in a 16S central pair-associated complex. Motility defects associated with cpc1 alleles in vivo are partially rescued in vitro by reactivation of axonemes or cell models in saturating concentrations of ATP; thus the Cpc1 complex is essential for maintaining normal ATP concentrations in the flagellum.
p120 catenin is required for morphogenetic movements involved in the formation of the eyes and the craniofacial skeleton inXenopus
Authors: Ciesiolka Malgorzata , Delvaeye Mieke , Van Imschoot Griet , Verschuere Veerle , McCrea Pierre , van Roy Frans , Vleminckx Kris
Published: August 2004

During Xenopus development, p120 transcripts are enriched in highly morphogenetic tissues. We addressed the developmental function of p120 by knockdown experiments and by expressing E-cadherin mutants unable to bind p120. This resulted in defective eye formation and provoked malformations in the craniofacial cartilage structures, derivatives of the cranial neural crest cells. Closer inspection showed that p120 depletion impaired evagination of the optic vesicles and migration of cranial neural crest cells from the neural tube into the branchial arches. These morphogenetic processes were also affected by p120-uncoupled cadherins or E-cadherin containing a deletion of the juxtamembrane domain. Irrespective of the manipulation that caused the malformations, coexpression of dominant-negative forms of either Rac1 or LIM kinase rescued the phenotypes. Wild-type RhoA and constitutively active Rho kinase caused partial rescue. Our results indicate that, in contrast to invertebrates, p120 is an essential factor for vertebrate development and an adequate balance between cadherin activity and cytoskeletal condition is critical for correct morphogenetic movements.
Intentional Topiramate Ingestion in an Adolescent Female
Authors: Chung Allison M , Reed Michael D
Published: August 2004

OBJECTIVE:

To describe an intentional topiramate ingestion by an adolescent and warn of the potential for topiramate abuse.

CASE SUMMARY:

A 17-year-old female intentionally ingested approximately eight 100-mg topiramate tablets for the purpose of “getting high.” Soon after ingestion, she was found at school obtunded and nonresponsive. Upon transfer to the emergency department, she became combative and aggressive with evolving neurologic abnormalities including incoherence, confusion, disorientation, and significant speech impairments including echolalia. Approximately 24 hours after ingestion, the patient had completely recovered without requiring specific treatment or experiencing sequelae.

DISCUSSION:

The clinical effects following acute topiramate intoxication appear consistent with the drug's known pharmacologic properties. There are few other reports of topiramate ingestions and most cases have had mild outcomes.

CONCLUSIONS:

Due to the multifactorial effects topiramate may have upon the central nervous system and its anorectic effect, abuse of this drug by adolescents should be considered upon presentation of an adolescent with mental status changes.
Nonestrogen Treatment Modalities for Vasomotor Symptoms Associated with Menopause
Authors: Fugate Susan E , Church Chelsea O
Published: August 2004

OBJECTIVE:

To systematically review the literature regarding the efficacy and safety of nonestrogen treatments for menopause-associated vasomotor symptoms not due to cancer or chemotherapy.

DATA SOURCES:

Pertinent literature and clinical studies were identified by searching MEDLINE (1966–February 2004) and EMBASE (1959–February 2004) using the key search terms vasomotor symptoms, hot flashes, and menopause. Bibliographies of relevant articles were reviewed for additional references.

STUDY SELECTION AND DATA EXTRACTION:

English-language articles reporting efficacy and safety of nonestrogen treatment modalities for perimenopausal and postmenopausal vasomotor symptoms were evaluated. All articles identified from the data sources were evaluated, and all information deemed relevant was included. Emphasis was placed on randomized, double-blind, placebo-controlled clinical trials, as these provide the best efficacy and safety data. Studies evaluating treatment of vasomotor symptoms from other causes, such as cancer or chemotherapy, were excluded.

DATA SYNTHESIS:

Prescription medications reviewed for efficacy and safety in postmenopausal vasomotor symptoms include clonidine hydrochloride, danazol, gabapentin, methyldopa, mirtazapine, progestins, propranolol hydrochloride, selective serotonin-reuptake inhibitors (SSRIs), and venlafaxine. Nonprescription therapies reviewed include black cohosh, dong quai, evening primrose oil, physical activity, phytoestrogens, and red clover.

CONCLUSIONS:

According to this systematic literature review, postmenopausal vasomotor treatments that have been shown to be safe and effective in short-term use include black cohosh, exercise, gabapentin, medroxyprogesterone acetate, SSRIs (ie, paroxetine hydrochloride), and soy protein. Initial, small reports are suggestive for efficacy in menopausal vasomotor symptoms with megestrol acetate and venlafaxine.
Use of Single-Dose Rasburicase in an Obese Female
Authors: Arnold Tamra M , Reuter Jameson P , Delman Bryna S , Shanholtz Carl B
Published: August 2004

OBJECTIVE:

To report the use of single-dose rasburicase in an obese patient.

CASE SUMMARY:

A 53-year-old obese African American woman weighing 136 kg (ideal body weight [IBW] 55 kg) with new-onset chronic myelomonocytic leukemia in leukocytic blast crisis was treated with hydroxyurea 5 g daily. In addition, she received allopurinol 300 mg daily for prevention of tumor lysis syndrome (TLS). The following day, allopurinol was discontinued and rasburicase was administered at a dose of 0.2 mg/kg of IBW for a serum uric acid level of 11.9 mg/dL. The patient's serum uric acid level decreased to 1.9 mg/dL 48 hours after a single dose.

DISCUSSION:

Rasburicase is indicated for the initial management of elevated plasma uric acid levels in patients with hematologic and solid tumor malignancies who are at risk for TLS. This case is unique because the patient received one dose of rasburicase followed by allopurinol rather than 5 daily doses of rasburicase. Additionally, the dose was based on IBW rather than actual body weight. Efficacy of this approach is apparent from the uric acid levels and the lack of hemodialysis requirements.

CONCLUSIONS:

A single dose of rasburicase (based on IBW) followed by allopurinol can effectively prevent TLS based on serum uric acid concentration. This approach resulted in a substantial cost savings.
Direct Multiplex Assay of Lysosomal Enzymes in Dried Blood Spots for Newborn Screening
Authors: Li Yijun , Scott C Ronald , Chamoles Nestor A , Ghavami Ahmad , Pinto B Mario , Turecek Frantisek , Gelb Michael H
Published: August 2004

Background: Newborn screening for deficiency in the lysosomal enzymes that cause Fabry, Gaucher, Krabbe, Niemann–Pick A/B, and Pompe diseases is warranted because treatment for these syndromes is now available or anticipated in the near feature. We describe a multiplex screening method for all five lysosomal enzymes that uses newborn-screening cards containing dried blood spots as the enzyme source.

Methods: We used a cassette of substrates and internal standards to directly quantify the enzymatic activities, and tandem mass spectrometry for enzymatic product detection. Rehydrated dried blood spots were incubated with the enzyme substrates. We used liquid-liquid extraction followed by solid-phase extraction with silica gel to remove buffer components. Acarbose served as inhibitor of an interfering acid α-glucosidase present in neutrophils, which allowed the lysosomal enzyme implicated in Pompe disease to be selectively analyzed.

Results: We analyzed dried blood spots from 5 patients with Gaucher, 5 with Niemann–Pick A/B, 11 with Pompe, 5 with Fabry, and 12 with Krabbe disease, and in all cases the enzyme activities were below the minimum activities measured in a collection of heterozygous carriers and healthy noncarrier individuals. The enzyme activities measured in 5–9 heterozygous carriers were approximately one-half those measured with 15–32 healthy individuals, but there was partial overlap of each condition between the data sets for carriers and healthy individuals.

Conclusion: For all five diseases, the affected individuals were detected. The assay can be readily automated, and the anticipated reagent and supply costs are well within the budget limits of newborn-screening centers.
A Comparison Study of Hair Examination Methodologies
Authors: Kolowski JC , Petraco N , Wallace MM , De Forest PR , Prinz M
Published: August 2004

A study was conducted to investigate the accuracy between two methods of hair analysis: PCR-STR DNA analysis and microscopic comparison analysis. Standard sets of pubic hairs were collected from volunteers, and unknown sets were generated from these samples. Three out of five (60%) of the hairs analyzed produced full DNA profiles that were correctly matched to the standard sets. DNA analysis was inconclusive (partial or no DNA profile) for two out of five (40%) of the samples. In contrast, the microscopic comparison analysis correctly matched four out of five (80%) of the samples to the standard sets but mis-identified one out of five (20%) of the samples. These results reinforce the practice of preliminary microscopic hair examination in narrowing down a set of hairs for DNA analysis. Microscopic comparison analysis is sufficiently reliable to remain a rapid and inexpensive method for forensic hair analysis.
Visualization and Quantification of Air Embolism Structure by Processing Postmortem MSCT Data
Authors: Jackowski C , Thali M , Sonnenschein M , Aghayev E , Yen K , Dirnhofer R , Vock P
Published: August 2004

Venous air embolism (VAE) is an often occurring forensic finding in cases of injury to the head and neck. Whenever found, it has to be appraised in its relation to the cause of death. While visualization and quantification is difficult at traditional autopsy, Magnetic Resonance Imaging (MRI) and Computed Tomography (CT) offer a new potential in the diagnosis of VAE. This paper reports the findings of VAE in four cases of massive head injury examined postmortem by Multislice Computed Tomography (MSCT) prior to autopsy. MSCT data of the thorax were processed using 3D air structure reconstruction software to visualize air embolism within the vascular system. Quantification of VAE was done by multiplying air containing areas on axial 2D images by their reconstruction intervals and then by summarizing the air volumes. Excellent 3D visualization of the air within the vascular system was obtained in all cases, and the intravascular gas volume was quantified.
Dating of Ballpoint Pen Ink
Authors: Hofer R
Published: August 2004

In this paper we describe a case in which a cash book, dated of the year 2000, was sent to the Forensic Science Division, Document Laboratory, Zurich Canton Police in March 2003. The questioned document was a list of 29 pages containing a consecutive handwritten numbering and dated entries (payments) made of blue ballpoint pen ink. By definition, a cash book has to be written by hand and the entries have to be made daily. The questioned document was suspected to have been written within a short period (e.g., a few hours) and backdated. The document lab was asked to determine the date of the entries of the questioned list. On one hand, we were asked, if the cash book had been kept on account consecutively during the period of one year, and on the other hand, the judges were interested in knowing, if the document could have been written in 2003 and back dated to 2000. To answer these questions, the document was examined for latent elements by electrostatic detection device (ESDA). The relative dating of the entries was performed by the quantification of the ballpoint pen ink dyes and their degradation products using HPLC. Results show that it is possible to determine the relative age of entries written by ballpoint pen within a relatively short time scale, if storage and supporting material of the different samples are the same or at least similar.
Precision and Reliability of Pulp/Tooth Area Ratio (RA) of Second Molar as Indicator of Adult Age
Authors: Cameriere R , Ferrante L , Cingolani M
Published: August 2004

This paper discusses a method for assessing adult age based on the relationship between age and measurement of the pulp/tooth area ratio (RA) of the second molar teeth. The sample consisted of 312 Italian white Caucasian patients (135 men, 177 women) aged between 14 and 24 years. Orthopantomograph samples (OPGs) were analyzed by the same observer and calibrated by κ statistics. Probabilities of an individual being older than 18 years of age (adult age) were derived using measurements of the ratio pulp/tooth area (RA) of the second molar teeth and the dichotomized stages of third molar development (Tm) as explanatory variables. These results were exploited to establish a threshold value to assign an individual to juvenile or adult age. A cut-off value of RA* = 0.088 was applied if Tm = 0 and RA* = 0.097 if Tm = 1. The sensitivity of this test was 91% and its specificity 94.5%. The proportion of individuals with correct classifications was 92%. Erratum to this paper appears in 49(6).
A SingleIn VivoExposure to Cocaine Abolishes Endocannabinoid-Mediated Long-Term Depression in the Nucleus Accumbens
Authors: Fourgeaud Lawrence , Mato Susana , Bouchet Delphine , Hémar Agnès , Worley Paul F. , Manzoni Olivier J.
Published: August 2004

In the nucleus accumbens (NAc), a key structure to the effects of all addictive drugs, presynaptic cannabinoid CB1 receptors (CB1Rs) and postsynaptic metabotropic glutamate 5 receptors (mGluR5s) are the principal effectors of endocannabinoid (eCB)-mediated retrograde long-term depression (LTD) (eCB-LTD) at the prefrontal cortex-NAc synapses. Both CB1R and mGluR5 are involved in cocaine-related behaviors; however, the impact ofin vivococaine exposure on eCB-mediated retrograde synaptic plasticity remains unknown. Electrophysiological and biochemical approaches were used, and we report that a singlein vivococaine administration abolishes eCB-LTD. This effect of cocaine was not present in D1dopamine receptor (D1R) -/- mice and was prevented when cocaine was coadministered with the selective D1R antagonist 8-chloro-2,3,4,5-tetrahydro-3-5-1h-3-benzazepin-7-ol (0.5 mg/kg) or with the NMDA receptor (NMDAR) blocker (+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate (1 mg/kg), suggesting the involvement of D1R and NMDAR. We found that the cocaine-induced blockade of retrograde signaling was correlated with enhanced expression levels of Homer scaffolding proteins containing the coiled-coil domain and accompanied by a strong reduction of mGluR5 surface expression. The results suggest that cocaine-induced loss of eCB retrograde signaling is caused by a reduction in the ability of mGluR5 to translate anterograde glutamate transmission into retrograde eCB signaling.
A Morphological Correlate of Synaptic Scaling in Visual Cortex
Authors: Wallace Wes , Bear Mark F.
Published: August 2004

We studied the response of dendritic spines in the thalamic-recipient zone of rat visual cortex to simple manipulations of the visual environment. We measured the morphologies of a total of 3824 spines located on the basal dendrites of 60 layer 3 pyramidal cells. As expected from previous studies, we found a significantly lower spine density in dark-reared animals at postnatal day 30 (P30) compared with light-reared controls. Additional analysis revealed that the spines in dark-reared animals were significantly shorter and more bulbous than in light-reared animals. When these two results were combined, we found that the total synaptic area per unit length of dendrite was conserved, compatible with the phenomenon of “synaptic scaling.” We also found that the increase in average spine head diameter is reversed by 10 d of light exposure (starting at P20), but surprisingly, the decrease in spine density is not. Thus, not all effects of dark rearing can be reversed by subsequent visual experience, even when the experience occurs during the third postnatal week.
Striatal Neuron Differentiation from Neurosphere-Expanded Progenitors Depends onGsh2Expression
Authors: Jensen Josephine B. , Björklund Anders , Parmar Malin
Published: August 2004

Neural stem and progenitor cells from the embryonic forebrain can be expanded under growth factor stimulationin vitro, either as free-floating aggregates called neurospheres or as attached monolayer cultures. We have previously shown that despite the maintenance of important regulatory genes such asGsh2, in vitroexpansion of cells from the lateral ganglion eminence (LGE) restricts their differentiation potential. Specifically, their ability to differentiate into striatal projection neurons is compromised. It is not clear whether this restriction is caused by loss of progenitors with the ability to generate striatal projection neurons or whether the restricted differentiation potential is caused by factors lacking duringin vitrodifferentiation. To address this, we have set up anin vitrosystem, in which expanded LGE-derived cells are differentiated in coculture with primary cells isolated from different regions of the embryonic brain. We provide evidence that the primary cells supply the expanded cells with contact-mediated region-specific developmental cues. Neurosphere-expanded LGE progenitors can, when presented with these cues, differentiate into neurons with characteristics of striatal projection neurons. Furthermore, we show that the ability of the expanded LGE cells to respond to the developmental cues presented by the primary cells depends on the maintained expression of Gsh2 in the expanded cells.
Retrieving Memories via Internal Context Requires the Hippocampus
Authors: Kennedy Pamela J. , Shapiro Matthew L.
Published: August 2004

Episodic memory encodes the unique contexts of events so that people can remember the details of an experience when cued by only a subset of event features (Tulving, 1972). In humans, the hippocampus is crucial for this kind of memory (Scoville and Milner, 1957; Vargha-Khadem et al., 1997). The present study tested whether the hippocampus was required for nonspatial, context-dependent memory retrieval in rats that were trained in a constant external environment to approach different nonspatial goal objects depending on their current internal motivational state (hunger or thirst). The rats learned to reliably approach the correct goal and thus used internal context to guide associative memory retrieval. Both fornix transection and selective neurotoxic hippocampal lesions severely impaired memory performance, but cue and motivational discrimination, as well as stimulus-reward associations, were preserved. The findings suggest that the hippocampus is required for using internal contextual information for flexible associative memory retrieval.