bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization and Intracellular Localization of the Epstein-Barr Virus Protein BFLF2: Interactions with BFRF1 and with the Nuclear Lamina
Authors: Gonnella Roberta , Farina Antonella , Santarelli Roberta , Raffa Salvatore , Feederle Regina , Bei Roberto , Granato Marisa , Modesti Andrea , Frati Luigi , Delecluse Henri-Jacques , Torrisi Maria Rosaria , Angeloni Antonio , Faggioni Alberto
Published: February 2005

We have reported in the accompanying paper that the BFRF1 protein of Epstein-Barr virus (EBV) is important for efficient primary viral envelopment and egress (A. Farina, R. Feederle, S. Raffa, R. Gonnella, R. Santarelli, L. Frati, A. Angeloni, M. R. Torrisi, A. Faggioni, and H.-J. Delecluse, J. Virol. 79:3703-3712). Here we describe the characterization of the product of the EBV BFLF2 gene, which belongs to a family of conserved herpesviral genes which include the UL31 genes of herpes simplex virus and of pseudorabies virus and whose products are known to interact with UL34, the positional homolog of BFRF1. BFLF2 is an early transcript and is expressed in a variety of cell lines upon EBV lytic cycle activation. Western blotting of purified virion preparations showed that BFLF2 is a component of intracellular virions but is absent from mature extracellular virions. Coimmunoprecipitation experiments indicated that BFLF2 interacts with BFRF1, which was confirmed by immunofluorescence confocal microscopy showing that the two proteins colocalize on the nuclear membrane not only upon cotransfection in epithelial cells but also during viral replication. In cells carrying an EBV mutant with the BFRF1 gene deleted (293-BFRF1-KO cells) BFLF2 expression was low, and it was restored to wild-type levels upon treatment of the cells with the proteasome inhibitor MG132. Furthermore, recomplementing the 293-BFRF1-KO cells by BFRF1 transfection restored BFLF2 expression to the wild-type level. In addition, when expressed alone BFLF2 was localized diffusely inside the nucleus, whereas in the presence of BFRF1 the two proteins colocalized at the nuclear rim. Finally, 293 epithelial cells transfected with either protein or cotransfected were analyzed by electron microscopy to investigate potential alterations in the morphology of the nuclear membrane. The ultrastructural analysis revealed that (i) BFRF1 caused duplications of the nuclear membrane, similar to those reported to occur during the course of herpesviral replication, and (ii) while BFLF2 alone did not cause any apparent alteration, coexpression of the two proteins dramatically induced profound convolutions of the duplicated nuclear membrane. Both biochemical and morphological analysis showed association of the BFRF1-BFLF2 complex with a component of the nuclear lamina, lamin B. Taken together, these results and those of the accompanying paper (Farina et al., J. Virol. 79:3703-3712) indicate an important role of BFRF1 and BFLF2 in the early steps of EBV maturation at the nuclear membrane.
Structural Analysis of Tobacco Etch Potyvirus HC-Pro Oligomers Involved in Aphid Transmission
Authors: Ruiz-Ferrer Virginia , Boskovic Jasminka , Alfonso Carlos , Rivas Germán , Llorca Oscar , López-Abella Dionisio , López-Moya Juan José
Published: February 2005

Oligomeric forms of the HC-Pro protein of the tobacco etch potyvirus (TEV) have been analyzed by analytical ultracentrifugation and single-particle electron microscopy combined with three-dimensional (3D) reconstruction. Highly purified HC-Pro protein was obtained from plants infected with TEV by using a modified version of the virus that incorporates a histidine tag at the HC-Pro N terminus (hisHC-Pro). The purified protein retained a high biological activity in solution when tested for aphid transmission. Sedimentation equilibrium showed that the hisHC-Pro preparations were heterogenous in size. Sedimentation velocity confirmed the previous observation and revealed that the active protein solution contained several sedimenting species compatible with dimers, tetramers, hexamers, and octamers of the protein. Electron microscopy fields of purified protein showed particles of different sizes and shapes. The reconstructed 3D structures suggested that the observed particles could correspond to dimeric, tetrameric, and hexameric forms of the protein. A model of the interactions required for oligomerization of the HC-Pro of potyviruses is proposed.
The E2 Glycoprotein of Classical Swine Fever Virus Is a Virulence Determinant in Swine
Authors: Risatti G. R. , Borca M. V. , Kutish G. F. , Lu Z. , Holinka L. G. , French R. A. , Tulman E. R. , Rock D. L.
Published: February 2005

To identify genetic determinants of classical swine fever virus (CSFV) virulence and host range, chimeras of the highly pathogenic Brescia strain and the attenuated vaccine strain CS were constructed and evaluated for viral virulence in swine. Upon initial screening, only chimeras 138.8v and 337.14v, the only chimeras containing the E2 glycoprotein of CS, were attenuated in swine despite exhibiting unaltered growth characteristics in primary porcine macrophage cell cultures. Additional viral chimeras were constructed to confirm the role of E2 in virulence. Chimeric virus 319.1v, which contained only the CS E2 glycoprotein in the Brescia background, was markedly attenuated in pigs, exhibiting significantly decreased virus replication in tonsils, a transient viremia, limited generalization of infection, and decreased virus shedding. Chimeras encoding all Brescia structural proteins in a CS genetic background remained attenuated, indicating that additional mutations outside the structural region are important for CS vaccine virus attenuation. These results demonstrate that CS E2 alone is sufficient for attenuating Brescia, indicating a significant role for the CSFV E2 glycoprotein in swine virulence.
Exogenous ACE2 Expression Allows Refractory Cell Lines To Support Severe Acute Respiratory Syndrome Coronavirus Replication
Authors: Mossel Eric C. , Huang Cheng , Narayanan Krishna , Makino Shinji , Tesh Robert B. , Peters C. J.
Published: February 2005

Of 30 cell lines and primary cells examined, productive severe acute respiratory syndrome coronavirus (Urbani strain) (SARS-CoV) infection after low-multiplicity inoculation was detected in only six: three African green monkey kidney epithelial cell lines (Vero, Vero E6, and MA104), a human colon epithelial line (CaCo-2), a porcine kidney epithelial line [PK(15)], and mink lung epithelial cells (Mv 1 Lu). SARS-CoV produced a lytic infection in Vero, Vero E6, and MA104 cells, but there was no visible cytopathic effect in Caco-2, Mv 1 Lu, or PK(15) cells. Multistep growth kinetics were identical in Vero E6 and MA104 cells, with maximum titer reached 24 h postinoculation (hpi). Virus titer was maximal 96 hpi in CaCo-2 cells, and virus was continually produced from infected CaCo-2 cells for at least 6 weeks after infection. CaCo-2 was the only human cell type of 13 tested that supported efficient SARS-CoV replication. Expression of the SARS-CoV receptor, angiotensin-converting enzyme 2 (ACE2), resulted in SARS-CoV replication in all refractory cell lines examined. Titers achieved were variable and dependent upon the method of ACE2 expression.
Novel Polyomavirus Detected in the Feces of a Chimpanzee by Nested Broad-Spectrum PCR
Authors: Johne Reimar , Enderlein Dirk , Nieper Hermann , Müller Hermann
Published: February 2005

In order to screen for new polyomaviruses in samples derived from various animal species, degenerated PCR primer pairs were constructed. By using a nested PCR protocol, the sensitive detection of nine different polyomavirus genomes was demonstrated. The screening of field samples revealed the presence of a new polyomavirus, tentatively designated chimpanzee polyomavirus (ChPyV), in the feces of a juvenile chimpanzee (
Pan troglodytes
). Analysis of the region encoding the major capsid protein VP1 revealed a unique insertion in the EF loop of the protein and showed that ChPyV is a distinct virus related to the monkey polyomavirus B-lymphotropic polyomavirus and the human polyomavirus JC polyomavirus.
Characterization and functional expression in yeast of a cDNA encoding NADP-dependent malic enzyme from castor oil seed
Authors: Shearer Heather L , Dennis David T
Published: February 2005

A cDNA encoding NADP-dependent malic enzyme was isolated from castor bean (Ricinus communis L.) endosperm. The 2362-bp cDNA encodes a 641 amino acid protein, which is predicted to have a 50 amino acid chloroplast transit peptide. Southern blotting revealed a single copy of the gene. Recombinant protein expression confirmed that the cDNA encodes a NADP-dependent malic enzyme; both Western blotting and enzyme assays detected NADP-dependent malic enzyme. Removal of the transit peptide in the recombinant protein did not affect the pH optimum of the enzyme, but it did result in an increased specific activity.Key words: Ricinus communis, NADP-dependent malic enzyme (NADP-ME), fatty acid biosynthesis, leucoplast, recombinant protein expression, transit peptide.
PROTEIN FOLDING ON THE HEXAGONAL LATTICE IN THE HP MODEL
Authors: JIANG MINGHUI , ZHU BINHAI
Published: February 2005

In this paper, we introduce the 2D hexagonal lattice as a biologically meaningful alternative to the standard square lattice for the study of protein folding in the HP model. We show that the hexagonal lattice alleviates the "sharp turn" problem and models certain aspects of the protein secondary structure more realistically. We present a ⅙-approximation and a clustering heuristic for protein folding on the hexagonal lattice. In addition to these two algorithms, we also implement a Monte Carlo Metropolis algorithm and a branch-and-bound partial enumeration algorithm, and conduct experiments to compare their effectiveness.
Pathogen Testing and Certification ofVitisandPrunusSpecies
Authors: Rowhani Adib , Uyemoto Jerry K. , Golino Deborah A. , Martelli Giovanni P.
Published: February 2005

Strategies to screen horticultural crops for graft-transmissible agents, particularly viruses and phytoplasmas, have advanced substantially over the past decade. Tests used for Vitis and Prunus are reviewed in detail, including both biological indexing procedures and laboratory-based assays. Despite advances in laboratory molecular-based detection techniques, a strong case is presented for the continued use of slower biological tests in programs requiring high levels of confidence in detection of pathogens that must be excluded from valuable germplasm.
Principles of Plant Health Management for Ornamental Plants
Authors: Daughtrey Margery L. , Benson D. Michael
Published: February 2005

Economic, environmental, and technological influences complicate the task of achieving disease-free products in the ornamentals industry. Integrated pest management (IPM) is a cornerstone of floriculture and nursery crop production: strategies include sanitation, clean stock, host resistance, and control through biological, cultural, environmental, chemical, and regulatory means. Sanitation measures and cultural controls must keep pace with new production technologies. Clean stock programs are used for many crops that are propagated vegetatively. Breeding, selection, and biotechnology provide crops resistant to pathogens. Offshore production for economic competitiveness can introduce pathogens that make regulatory programs necessary. New biocontrol and chemical products continue to improve control while meeting the requirement for minimal environmental impact. Continual introduction of new crops and new production technologies creates new opportunities for pathogens to exploit, such that new disease management tactics must be discovered and old ones rediscovered to achieve optimum health management for ornamentals.
Plant Pathology and RNAi: A Brief History
Authors: Lindbo John A. , Dougherty William G.
Published: February 2005

This article describes the discovery of RNA-activated sequence-specific RNA degradation, a phenomenon now referred to as RNA silencing or RNA interference (RNAi). From 1992 to 1996, a series of articles were published on virus resistant transgenic plants expressing either translatable or nontranslatable versions of the coat protein gene of Tobacco etch virus (TEV). Certain transgenic plant lines were resistant to TEV but not to closely related viruses. In these plants a surprising correlation was observed: Transgenic plant lines with the highest degree of TEV resistance had actively transcribed transgenes but low steady-state levels of transgene RNA. Molecular analysis of these transgenic plants demonstrated the existence of a cellular-based, sequence-specific, posttranscriptional RNA-degradation system that was programmed by the transgene-encoded RNA sequence. This RNA-degradation activity specifically targeted both the transgene RNA and TEV (viral) RNA for degradation and was the first description of RNA-mediated gene silencing.
Mechanism and regulation of human intestinal niacin uptake
Authors: Nabokina Svetlana M. , Kashyap Moti L. , Said Hamid M.
Published: February 2005

The mechanism of uptake of dietary niacin (nicotinic acid) by intestinal epithelial cells is not well understood, and nothing is known about regulation of the uptake process. In this investigation, we used human-derived intestinal epithelial Caco-2 cells and purified intestinal brush-border membrane vesicles (BBMVs) isolated from human organ donors to assess niacin uptake. Our findings show niacin uptake by Caco-2 cells to be 1) temperature and energy dependent; 2) Na+ independent, but highly dependent on extracellular acidic pH; 3) saturable as a function of concentration, with an apparent Km of 0.53 ± 0.08 μM; 4) severely inhibited by the membrane-impermeable sulfhydryl group of reagents; and 5) highly specific for niacin but not affected by monocarboxylic acids. A marked trans stimulation in [3H]niacin efflux from preloaded Caco-2 cells by unlabeled niacin in the incubation buffer was also observed. These findings suggest the involvement of a specialized, pH-dependent, carrier-mediated mechanism for human intestinal niacin uptake. This suggestion was confirmed in studies with native human intestinal BBMVs. We also examined possible regulation of niacin uptake by Caco-2 cells via specific intracellular regulatory pathways. The results show that while the PKA-, PKC-, and Ca2+/calmodulin-mediated regulatory pathways play no role in regulating niacin uptake, a role for a protein tyrosine kinase (PTK)-mediated pathway is apparent. The results of these studies show for the first time the existence of a specialized, acidic pH-dependent, carrier-mediated system of niacin uptake by human intestinal epithelial cells that operates at the micromolar (physiological) range of niacin. The results also suggest the possible involvement of a PTK-mediated pathway in the regulation of niacin uptake.
Water and solute permeability of rat lung caveolae: high permeabilities explained by acyl chain unsaturation
Authors: Hill Warren G. , Almasri Eyad , Ruiz W. Giovanni , Apodaca Gerard , Zeidel Mark L.
Published: February 2005

Caveolae are invaginated membrane structures with high levels of cholesterol, sphingomyelin, and caveolin protein that are predicted to exist as liquid-ordered domains with low water permeability. We isolated a caveolae-enriched membrane fraction without detergents from rat lung and characterized its permeability properties to nonelectrolytes and protons. Membrane permeability to water was 2.85 ± 0.41 × 10−3 cm/s, a value 5–10 times higher than expected based on comparisons with other cholesterol and sphingolipid-enriched membranes. Permeabilities to urea, ammonia, and protons were measured and found to be moderately high for urea and ammonia at 8.85 ± 2.40 × 10−7and 6.84 ± 1.03 × 10−2 respectively and high for protons at 8.84 ± 3.06 × 10−2 cm/s. To examine whether caveolin or other integral membrane proteins were responsible for high permeabilities, liposomes designed to mimic the lipids of the inner and outer leaflets of the caveolar membrane were made. Osmotic water permeability to both liposome compositions were determined and a combined inner/outer leaflet water permeability was calculated and found to be close to that of native caveolae at 1.58 ± 1.1 × 10−3 cm/s. In caveolae, activation energy for water flux was high (19.4 kcal/mol) and water permeability was not inhibited by HgCl2; however, aquaporin 1 was detectable by immunoblotting. Immunostaining of rat lung with AQP1 and caveolin antisera revealed very low levels of colocalization. We conclude that aquaporin water channels do not contribute significantly to the observed water flux and that caveolae have relatively high water and solute permeabilities due to the high degree of unsaturation in their fatty acyl chains.
Is myosin light-chain phosphorylation a regulatory signal for the osmotic activation of the Na+-K+-2Cl− cotransporter?
Authors: Ciano-Oliveira Caterina Di , Lodyga Monika , Fan Lingzhi , Szászi Katalin , Hosoya Hiroshi , Rotstein Ori D. , Kapus András
Published: February 2005

Myosin light-chain (MLC) kinase (MLCK)-dependent increase in MLC phosphorylation has been proposed to be a key mediator of the hyperosmotic activation of the Na+-K+-2Cl− cotransporter (NKCC). To address this hypothesis and to assess whether MLC phosphorylation plays a signaling or permissive role in NKCC regulation, we used pharmacological and genetic means to manipulate MLCK, MLC phosphorylation, or myosin ATPase activity and followed the impact of these alterations on the hypertonic stimulation of NKCC in porcine kidney tubular LLC-PK1 epithelial cells. We found that the MLCK inhibitor ML-7 suppressed NKCC activity independently of MLC phosphorylation. Notably, ML-7 reduced both basal and hypertonically stimulated NKCC activity without influencing MLC phosphorylation under these conditions, and it inhibited NKCC activation by Cl− depletion, a treatment that did not increase MLC phosphorylation. Furthermore, prevention of the osmotically induced increase in MLC phosphorylation by viral induction of cells with a nonphosphorylatable, dominant negative MLC mutant (AA-MLC) did not affect the hypertonic activation of NKCC. Conversely, a constitutively active MLC mutant (DD-MLC) that mimics the diphosphorylated form neither stimulated isotonic nor potentiated hypertonic NKCC activity. Furthermore, a depolarization-induced increase in endogenous MLC phosphorylation failed to activate NKCC. However, complete abolition of basal MLC phosphorylation by K252a or the inhibition of myosin ATPase by blebbistatin significantly reduced the osmotic stimulation of NKCC without suppressing its basal or Cl− depletion-triggered activity. These results indicate that an increase in MLC phosphorylation is neither a sufficient nor a necessary signal to stimulate NKCC in tubular cells. However, basal myosin activity plays a permissive role in the optimal osmotic responsiveness of NKCC.
Depressed Responses of Facilitatory Synapses
Authors: Banitt Yoav , Martin Kevan A. C. , Segev Idan
Published: February 2005

We show that when temporal summation takes place, depression of postsynaptic responses may ensue when the underlying synaptic conductance change is constant or even facilitatory. We term this phenomenon “apparent depression.” Such apparent depression is most notable for slow synaptic conductance changes, for high frequency, and when the synapse is located at distal dendritic sites. We show that, when temporal summation ensues, the erroneous estimation of short-term synaptic plasticity arises partially from the conventional measurement of synaptic dynamics at postsynaptic potential peak time. This can be corrected when measuring overlapping synaptic responses at fixed intervals after stimulus time. Somatic voltage clamp also helps to partially correct for the apparent depression, but a good model of the neuron can do even better in providing a more accurate view of the underlying synaptic conductances.
Deliberative Democracy in Divided Societies
Authors: Dryzek John S.
Published: February 2005

For contemporary democratic theorists, democracy is largely a matter of deliberation. But the recent rise of deliberative democracy (in practice as well as theory) coincided with ever more prominent identity politics, sometimes in murderous form in deeply divided societies. This essay considers how deliberative democracy can process the toughest issues concerning mutually contradictory assertions of identity. After considering the alternative answers provided by agonists and consociational democrats, the author makes the case for a power-sharing state with attenuated sovereignty and a more engaged deliberative politics in a public sphere that is semidetached from the state and situated transnationally.
Open Adoption as a Family Form
Authors: Miall Charlene E. , March Karen
Published: February 2005

In this article, community assessments of support for three levels of open adoption are examined, including two types of mediated adoption, fully disclosed adoption, and confidential adoption. Combining telephone survey data from a Canada-wide random sample of 706 respondents with 82 qualitative interviews, the authors report on community assessments of these levels of openness and the relevance of these results for other survey research conducted on these issues. The authors also explore community assessments of the perceived advantages and disadvantages of open adoption and confidential adoption for adoptive parents, birthparents, and adopted children and how these may or may not reflect clinical assessments. Finally, the authors consider the implications of these results for clinical initiatives with members of the adoption triangle.
Evaluation of the Effects of the Aban Aya Youth Project in Reducing Violence among African American Adolescent Males Using Latent Class Growth Mixture Modeling Techniques
Authors: Segawa Eisuke , Ngwe Job E. , Li Yanhong , Flay Brian R. ,   Aban Aya Coinvestigators
Published: February 2005

This study employs growth mixture modeling techniques to evaluate the preventive effects of the Aban Aya Youth Project in reducing the rate of growth of violence among African American adolescent males (N = 552). Results suggest three distinct classes of participants: high risk (34%), medium risk (54%), and low risk (12%) based on both the participants’ initial violence scores and their growth of violence over time. Results further show significant effects (almost 3 times as large as the effect found in the regular one-class analysis) for the high-risk class but not for the medium- or low-risk classes.
Mutational study of sapovirus expression in insect cells
Authors: Hansman Grant S , Katayama Kazuhiko , Oka Tomoichiro , Natori Katsuro , Takeda Naokazu
Published: February 2005

Human sapovirus (SaV), an agent of human gastroenteritis, cannot be grown in cell culture, but expression of the recombinant capsid protein (rVP1) in a baculovirus expression system results in the formation of virus-like particles (VLPs). In this study we compared the time-course expression of two different SaV rVP1 constructs. One construct had the native sequence (Wt construct), whereas the other had two nucleotide point mutations in which one mutation caused an amino acid substitution and one was silent (MEG-1076 construct). While both constructs formed VLPs morphologically similar to native SaV, Northern blot analysis indicated that the MEG-1076 rVP1 mRNA had increased steady-state levels. Furthermore, Western blot analysis and an antigen enzyme-linked immunosorbent assay showed that the MEG-1076 construct had increased expression levels of rVP1 and yields of VLPs. Interestingly, the position of the mutated residue was strictly conserved residue among other human SaV strains, suggesting an important role for rVP1 expression.
Inhibition of IL-18 reduces myeloperoxidase activity and prevents edema in intestine following alcohol and burn injury
Authors: Rana Shadab N , Li Xiaoling , Chaudry Irshad H , Bland Kirby I , Choudhry Mashkoor A
Published: February 2005

Previous studies have shown that alcohol (EtOH) ingestion before burn injury impaired intestinal barrier and immune function. This study determined whether EtOH and burn injury up-regulate interleukin (IL)-18 and whether IL-18 up-regulation following EtOH and burn injury is a cause for neutrophilrecruitment and increased intestinal edema. Rats (250 g) were gavaged with EtOH to achieve a blood EtOH level in the range of 100 mg/dL prior to burn or sham injury (25% total body surface area). A group of rats was treated with Ac-YVAD-CHO (5 mg/kg), an inhibitor of caspase-1 (an enzyme that converts pro-IL-18, an inactive form of IL-18, to mature IL-18), at the time of injury. One day after injury, rats were killed. IL-18 production was determined in circulation and in the supernatants harvested from spleen, mesenteric lymph nodes, and Peyer's patch cell cultures as well as in intestinal tissue homogenates. Neutrophil accumulation in intestine was determined by measuring myeloperoxidase (MPO) activity. We found a significant increase in IL-18 levels in the lymphoid cell supernatants and intestinal tissue homogenates obtained from EtOH and burn-injured rats compared with the rats receiving burn or sham injury. This was accompanied by an increase in intestinal MPO and edema. No demonstrable change in intestinal morphology was observed in any group. Treatment of rats with caspase-1 inhibitor significantly attenuated the increase in IL-18 levels and intestinal MPO activity in EtOH and burn-injured rats. Inhibition of IL-18 also prevented an increase in intestinal tissue water content. As MPO is considered an index of neutrophil infiltration, results presented in this manuscript collectively suggest that IL-18 up-regulation is likely to contribute to the increased neutrophil infiltration and edema in intestinal tissue observed following EtOH and burn injury.
Pro- and anti-inflammatory substances modulate expression of the leukotriene B4 receptor, BLT1, in human monocytes
Authors: Pettersson Annika , Sabirsh Alan , Bristulf Jesper , Kidd-Ljunggren Karin , Ljungberg Bengt , Owman Christer , Karlsson Ulf
Published: February 2005

The high-affinity leukotriene B4 (LTB4) receptor, BLT1, is a chemotactic receptor involved in inflammatory responses. In this study, we have explored the regulation of BLT1 expression in human monocytes by pro- and anti-inflammatory cytokines, lipopolysaccharide (LPS), and dexamethasone. We found that proinflammatory mediators, such as interferon-γ (IFN-γ), tumor necrosis factor-α, and LPS, down-regulated expression, whereas the anti-inflammatory cytokine, interleukin-10, and dexamethasone up-regulated BLT1 mRNA expression. The effect of IFN-γ on BLT1 mRNA expression was rapidly detectable (<4 h) and concentration-dependent (1–50 ng/ml) and seems to be exerted through a block in transcriptional activity. Alterations in mRNA expression were accompanied by changes in BLT1 surface expression, and receptor down-modulation following IFN-γ stimulation resulted in a diminished chemotactic response to LTB4. The regulation of BLT1 mRNA and receptor protein expression was similar to the regulation of the monocyte chemoattractant protein-1 chemokine receptor, CC chemokine recptor 2 (CCR2). Flow cytometric analysis of fresh peripheral blood cells revealed that classical (CD14++CD16–) monocytes express high levels of BLT1 and CCR2 and that both receptors are down-regulated on CD14+CD16+ monocytes. Apart from providing insight into the regulation of BLT1 in human monocytes, our results reveal a parallel expression and regulation of BLT1 and CCR2, which may help to understand monocyte trafficking during pathophysiological conditions.
An Alternative Approach to Characterize Nonlinear Site Effects
Authors: Zhang Ray Ruichong , Hartzell Stephen , Liang Jianwen , Hu Yuxian
Published: February 2005

This paper examines the rationale of a method of nonstationary data processing and analysis, referred to as the Hilbert-Huang transform (HHT), for its application to a recording-based approach in quantifying influences of soil nonlinearity in site response. In particular, this paper first summarizes symptoms of soil nonlinearity shown in earthquake recordings, reviews the Fourier-based approach to characterizing nonlinearity, and offers justifications for the HHT in addressing nonlinearity issues. This study then uses the HHT method to analyze synthetic data and recordings from the 1964 Niigata and 2001 Nisqually earthquakes. In doing so, the HHT-based site response is defined as the ratio of marginal Hilbert amplitude spectra, alternative to the Fourier-based response that is the ratio of Fourier amplitude spectra. With the Fourier-based approach in studies of site response as a reference, this study shows that the alternative HHT-based approach is effective in characterizing soil nonlinearity and nonlinear site response.
TREMOR: A Wireless MEMS Accelerograph for Dense Arrays
Authors: Evans John R. , Hamstra Robert H. , Kündig Christoph , Camina Patrick , Rogers John A.
Published: February 2005

The ability of a strong-motion network to resolve wavefields can be described on three axes: frequency, amplitude, and space. While the need for spatial resolution is apparent, for practical reasons that axis is often neglected. TREMOR is a MEMS-based accelerograph using wireless Internet to minimize lifecycle cost. TREMOR instruments can economically augment traditional ones, residing between them to improve spatial resolution. The TREMOR instrument described here has dynamic range of 96 dB between ±2 g, or 102 dB between ±4 g. It is linear to <1% of full scale (FS), with a response function effectively shaped electronically. We developed an economical, very low noise, accurate (<1%FS) temperature compensation method. Displacement is easily recovered to 10-cm accuracy at full bandwidth, and better with care. We deployed prototype instruments in Oakland, California, beginning in 1998, with 13 now at mean spacing of ∼3 km—one of the most densely instrumented urban centers in the United States. This array is among the quickest in returning (PGA, PGV, Sa) vectors to ShakeMap, ∼75 to 100 s. Some 13 events have been recorded. A ShakeMap and an example of spatial variability are shown. Extensive tests of the prototypes for a commercial instrument are described here and in a companion paper.
The Role of HOX Genes in Human Implantation
Authors: EUN KWON HYE , TAYLOR HUGH S.
Published: February 2005

Abstract: The endometrium undergoes an ordered process of differentiation leading to receptivity to embryonic implantation. HOX genes direct this development in a fashion similar to that in which they direct embryonic development, including development of the reproductive tract. HOXA10 and HOXA11 expression increases during the menstrual cycle, increasing drastically in the midluteal phase, at the time of implantation. This expression is regulated by sex steroid hormones. This expression is necessary for implantation of the blastocyst as demonstrated by the decreased implantation rates in women with altered HOX expression. HOX genes are markers of endometrial receptivity. The possibility of augmenting HOX gene expression with gene therapy to improve implantation has promise for the future.
The Vascular Cast of the Human Uterus: From Anatomy to Physiology
Authors: CICINELLI ETTORE , EINER‐JENSEN NIELS , GALANTINO PIETRO , ALFONSO RAFFAELLO , NICOLETTI ROBERTO
Published: February 2005

Abstract: The blood supply to the uterus originates mainly from the uterine artery. However, the uterine and ovarian arteries form anastomoses bilaterally. Controversy exists about the direction of the flow in the anastomoses and thus the origin of the arterial supply to the tube and tubal part of the uterus. A similar arcade is formed by the vaginal and uterine arteries. We have investigated the vascular border of supply between the uterine and ovarian arteries in postmenopausal women, which was positioned in the uterus 1‐2 cm from the tube. A similar result was found in younger, ovulating women. However, the border between the territories irrigated by the uterine and ovarian arteries differs between the follicular and luteal phase; more uterine tissue is perfused from the ovarian artery when a large follicle is present. This constitutes the first description of a functionally determined shift in the territorial divide of two vascular systems and has numerous practical implications. The venous system copies the arterial one with one major exception: some of the uterine veins join the ovarian outlet. The close contact between veins and arteries facilitates transfer of substances, thus forming semilocal systems of regulation; for example, the ovary locally influences the function of the ipsilateral tube and part of the uterus. From a therapeutic point of view, it has been documented that application of progesterone to the vagina creates high uterine concentrations due to local vascular transfer.
Myomas, Pregnancy Outcome, and In Vitro Fertilization
Authors: BULLETTI CARLO , DE ZIEGLER DOMINIQUE , LEVI SETTI PAOLO , CICINELLI ETTORE , POLLI VALERIA , STEFANETTI MARCO
Published: February 2005

Abstract: Uterine fibroids have been reported in 27% of infertile women, and 50% of women with unexplained infertility become pregnant after myomectomy. The age at which a first pregnancy occurs is increasing from the thirties to the forties. This increase and the recurrence rate of leiomyomas from 15 to 30% points to the effect of myomas on the infertility. Mechanisms by which myomas may cause infertility are abnormal uterine contractility, elongation of the uterine cavity, and distortion of uterine vascularization. Surgery may have beneficial or adverse effects without clear data on its effect on the assisted reproductive technology (ART) procedures. The present study was undertaken to establish the impact of surgical removal of myomas on fertility and infertility of patients undergoing ART procedures. Patients who underwent surgical removal of myomas before in vitro fertilization (Group A) had a cumulative success rate of 33% for one to three procedures (28 clinical pregnancies in 84 patients) and delivery rate of 25% (21 live births in 84 patients). Patients who underwent in vitro fertilization without previous surgery (Group B) had a 15% clinical pregnancy rate (13 pregnancies in 84 patients) (P < 0.05) and 12% delivery rate (10 deliveries in 84 pregnancies) (P < 0.05). Abortion rates were 7% (8 deliveries in 84 patients) and 4% (3 deliveries in 84 patients) in Groups A and B, respectively. This study confirms the beneficial effect of surgical removal of fibroids before undergoing ART procedures.
Assisted Reproductive Technologies in Europe Encompass Diverse and Complex Ethical Viewpoints: Issues to Be Considered in Reporting Research in Human Reproduction
Authors: BARLOW D H. , BEARD H K. , WILLIAMS A C.
Published: February 2005

Abstract: The many dimensions of the field of reproduction (clinical, scientific, social, ethical, legal) must be addressed by those with editorial responsibility for the major journals in the field. As the editorial team of the leading reproductive journal Human Reproduction, we have evolved processes that attempt to ensure a transparent but robust approach to complexities whose impact varies across international boundaries.
Genetic Basis of Endometriosis
Authors: BISCHOFF FARIDEH , SIMPSON JOE LEIGH
Published: February 2005

Abstract: Endometriosis is a complex gynecologic disorder that has long been recognized as showing heritable tendencies, with recurrence risks of 5‐7% for first‐degree relatives. Familial and epidemiologic studies support that this disease is a genetic disorder of polygenic/multifactorial inheritance. The current investigational challenge is to determine the number and location of causative genes. Recent advances in molecular technology make identification and elucidation of these genes now possible. In this review, we update previous communications in which we also reviewed heritability studies supporting polygenic/multifactorial inheritance, discuss the scientific basis of genomewide strategies for identifying causative genes, and identify potential candidate genes.
Endometriosis and Inflammation in Infertility
Authors: HALIS GÜLDEN , ARICI AYDIN
Published: February 2005

Abstract: A wealth of publications proposes that endometriosis and inflammation may have an unfavorable influence on fertility. A recent meta‐analysis of assisted reproductive technologies demonstrated that, once confounding factors are controlled for, the pregnancy rate in women with endometriosis is approximately 50% of the rate of women with tubal factor infertility. Peritoneal fluid of women with endometriosis contains elevated amounts of macrophages and their secreted products, such as growth factors, cytokines, and angiogenic factors. Because reproductive organs are bathed in and thus will be influenced by peritoneal fluid, these proinflammatory mediators would affect various aspects of reproduction in women with endometriosis. Advanced stages of endometriosis may have easily understandable factors, such as distortion of the anatomy, causing infertility. On the other hand, in minimal or mild endometriosis mechanisms underlying reproductive failure are subtle and remain controversial. Recent reports suggest that inflammatory factors play a role in this endometriosis‐associated reproductive failure. This review provides an overview of recent data on the effects of endometriosis‐associated inflammation on fertility.
Future Directions in Endometriosis Research
Authors: D'HOOGHE THOMAS M. , KYAMA C , DEBROCK S , MEULEMAN C , MWENDA JASON M.
Published: February 2005

Abstract: Endometriosis is an important gynecological disease, pathologically defined by the ectopic presence of both endometrial glands and stroma, and clinically associated with pelvic pain and infertility. Our current knowledge of the pathogenesis, pathophysiology of related infertility, and spontaneous evolution is still limited, although endometriosis has been described for many years. Future research in endometriosis needs to focus on pathogenesis studies in the baboon model and on the early interactions between endometrial and peritoneal cells in the pelvic cavity at the time of menstruation. Proteomic and genomic approaches are needed to detect potential differences between eutopic endometrium and myometrium in women with and without endometriosis. Immunomodulatory drugs inhibiting endometriosis‐associated pelvic inflammation may offer new medical treatment for endometriosis in the future.
Uterine Peristaltic Activity and the Development of Endometriosis
Authors: LEYENDECKER GERHARD , KUNZ GEORG , HERBERTZ MARK , BEIL DOLORES , HUPPERT PETER , MALL GERHARD , KISSLER STEPHAN , NOE MARION , WILDT LUDWIG
Published: February 2005

Abstract: Peristaltic activity of the nonpregnant uterus serves fundamental functions in the early process of reproduction, such as directed transport of spermatozoa into the tube ipsilateral to the dominant follicle, high fundal implantation of the embryo, and, possibly, retrograde menstruation. Hyperperistalsis of the uterus is significantly associated with the development of endometriosis and adenomyosis. In women with hyperperistalsis, fragments of basal endometrium are detached during menstruation and transported into the peritoneal cavity. Fragments of basal endometrium have, because of their equipment with estrogen and progesterone receptors and because of their ability to produce estrogen, an increased potential of implantation and proliferation, resulting in pelvic endometriosis. In addition, hyperperistalsis induces the proliferation of basal endometrium into myometrial dehiscencies. This results in endometriosis‐associated adenomyosis with a prevalence of approximately 90%. Adenomyosis results in impaired directed sperm transport and thus constitutes an important cause of sterility in women with endometriosis. Our own date and that from the literature strongly suggest that the principal mechanism of endometriosis/adenomyosis is the paracrine interference of endometrial estrogen with the cyclical endocrine control of archimyometrial peristalsis exerted by the ovary, thus resulting in hyperperistalsis.
Rab11 mediates post-Golgi trafficking of rhodopsin to the photosensitive apical membrane ofDrosophilaphotoreceptors
Authors: Satoh Akiko K. , O'Tousa Joseph E. , Ozaki Koichi , Ready Donald F.
Published: February 2005

In developing Drosophila photoreceptors, rhodopsin is trafficked to the rhabdomere, a specialized domain within the apical membrane surface. Rab11, a small GTPase implicated in membrane traffic, immunolocalizes to the trans-Golgi network, cytoplasmic vesicles and tubules, and the base of rhabdomeres. One hour after release from the endoplasmic reticulum, rhodopsin colocalizes with Rab11 in vesicles at the base of the rhabdomere. When Rab11 activity is reduced by three different genetic procedures, rhabdomere morphogenesis is inhibited and rhodopsin-bearing vesicles proliferate within the cytosol. Rab11 activity is also essential for development of MVB endosomal compartments; this is probably a secondary consequence of impaired rhabdomere development. Furthermore, Rab11 is required for transport of TRP, another rhabdomeric protein, and for development of specialized membrane structures within Garland cells. These results establish a role for Rab11 in the post-Golgi transport of rhodopsin and of other proteins to the rhabdomeric membranes of photoreceptors, and in analogous transport processes in other cells.
TheDrosophilatrithorax group protein Kismet facilitates an early step in transcriptional elongation by RNA Polymerase II
Authors: Srinivasan Shrividhya , Armstrong Jennifer A. , Deuring Renate , Dahlsveen Ina K. , McNeill Helen , Tamkun John W.
Published: February 2005

The Drosophila trithorax group gene kismet (kis)was identified in a screen for extragenic suppressors of Polycomb(Pc) and subsequently shown to play important roles in both segmentation and the determination of body segment identities. One of the two major proteins encoded by kis (KIS-L) is related to members of the SWI2/SNF2 and CHD families of ATP-dependent chromatin-remodeling factors. To clarify the role of KIS-L in gene expression, we examined its distribution on larval salivary gland polytene chromosomes. KIS-L is associated with virtually all sites of transcriptionally active chromatin in a pattern that largely overlaps that of RNA Polymerase II (Pol II). The levels of elongating Pol II and the elongation factors SPT6 and CHD1 are dramatically reduced on polytene chromosomes from kis mutant larvae. By contrast, the loss of KIS-L function does not affect the binding of PC to chromatin or the recruitment of Pol II to promoters. These data suggest that KIS-L facilitates an early step in transcriptional elongation by Pol II.
Quail-duck chimeras reveal spatiotemporal plasticity in molecular and histogenic programs of cranial feather development
Authors: Eames B. Frank , Schneider Richard A.
Published: February 2005

The avian feather complex represents a vivid example of how a developmental module composed of highly integrated molecular and histogenic programs can become rapidly elaborated during the course of evolution. Mechanisms that facilitate this evolutionary diversification may involve the maintenance of plasticity in developmental processes that underlie feather morphogenesis. Feathers arise as discrete buds of mesenchyme and epithelium, which are two embryonic tissues that respectively form dermis and epidermis of the integument. Epithelial-mesenchymal signaling interactions generate feather buds that are neatly arrayed in space and time. The dermis provides spatiotemporal patterning information to the epidermis but precise cellular and molecular mechanisms for generating species-specific differences in feather pattern remain obscure. In the present study, we exploit the quail-duck chimeric system to test the extent to which the dermis regulates the expression of genes required for feather development. Quail and duck have distinct feather patterns and divergent growth rates, and we exchange pre-migratory neural crest cells destined to form the craniofacial dermis between them. We find that donor dermis induces host epidermis to form feather buds according to the spatial pattern and timetable of the donor species by altering the expression of members and targets of the Bone Morphogenetic Protein, Sonic Hedgehog and Delta/Notch pathways. Overall, we demonstrate that there is a great deal of spatiotemporal plasticity inherent in the molecular and histogenic programs of feather development, a property that may have played a generative and regulatory role throughout the evolution of birds.
ROXY1, a member of the plant glutaredoxin family, is required for petal development inArabidopsis thaliana
Authors: Xing Shuping , Rosso Mario G. , Zachgo Sabine
Published: February 2005

We isolated three alleles of an Arabidopsis thaliana gene named ROXY1, which initiates a reduced number of petal primordia and exhibits abnormalities during further petal development. The defects are restricted to the second whorl of the flower and independent of organ identity. ROXY1 belongs to a subgroup of glutaredoxins that are specific for higher plants and we present data on the first characterization of a mutant from this large Arabidopsis gene family for which information is scarce. ROXY1 is predominantly expressed in tissues that give rise to new flower primordia, including petal precursor cells and petal primordia. Occasionally, filamentous organs with stigmatic structures are formed in the second whorl of the roxy1 mutant, indicative for an ectopic function of the class C gene AGAMOUS (AG). The function of ROXY1 in the negative regulation of AG is corroborated by premature and ectopic AG expression in roxy1-3 ap1-10 double mutants, as well as by enhanced first whorl carpeloidy in double mutants of roxy1 with repressors of AG, such as ap2 or lug. Glutaredoxins are oxidoreductases that oxidize or reduce conserved cysteine-containing motifs. Mutagenesis of conserved cysteines within the ROXY1 protein demonstrates the importance of cysteine 49 for its function. Our data demonstrate that, unexpectedly, a plant glutaredoxin is involved in flower development, probably by mediating post-translational modifications of target proteins required for normal petal organ initiation and morphogenesis.
Re-establishing the regenerative potential of central nervous system axons in postnatal mice
Authors: Cho Kin-Sang , Yang Liu , Lu Bin , Feng Ma Hong , Huang Xizhong , Pekny Milos , Chen Dong Feng
Published: February 2005

At a certain point in development, axons in the mammalian central nervous system lose their ability to regenerate after injury. Using the optic nerve model, we show that this growth failure coincides with two developmental events: the loss of Bcl-2 expression by neurons and the maturation of astrocytes. Before postnatal day 4, when astrocytes are immature, overexpression of Bcl-2 alone supported robust and rapid optic nerve regeneration over long distances, leading to innervation of brain targets by day 4 in mice. As astrocytes matured after postnatal day 4, axonal regeneration was inhibited in mice overexpressing Bcl-2. Concurrent induction of Bcl-2 and attenuation of reactive gliosis reversed the failure of CNS axonal re-elongation in postnatal mice and led to rapid axonal regeneration over long distances and reinnervation of the brain targets by a majority of severed optic nerve fibers up to 2 weeks of age. These results suggest that an early postnatal downregulation of Bcl-2 and post-traumatic reactive gliosis are two important elements of axon regenerative failure in the CNS.