bluuebunny/crossref_metadata_2025 huggingface dataset:
Ultraviolet radiation–induced injury, chemokines, and leukocyte recruitment: An amplification cycle triggering cutaneous lupus erythematosus
Authors: Meller Stephan , Winterberg Franziska , Gilliet Michel , Müller Anja , Lauceviciute Ingrida , Rieker Juliane , Neumann Norbert J. , Kubitza Robert , Gombert Michael , Bünemann Erich , Wiesner Ulrike , Franken‐Kunkel Petra , Kanzler Holger , Dieu‐Nosjean Marie‐Caroline , Amara Ali , Ruzicka Thomas , Lehmann Percy , Zlotnik Albert , Homey Bernhard
Published: May 2005

Objective

To investigate the activation and recruitment pathways of relevant leukocyte subsets during the initiation and amplification of cutaneous lupus erythematosus (LE).

Methods

Quantitative real‐time polymerase chain reaction was used to perform a comprehensive analysis of all known chemokines and their receptors in cutaneous LE lesions, and the cellular origin of these chemokines and receptors was determined using immunohistochemistry. Furthermore, cytokine‐ and ultraviolet (UV) light–mediated activation pathways of relevant chemokines were investigated in vitro and in vivo.

Results

In the present study, we identified the CXCR3 ligands CXCL9 (interferon‐γ [IFNγ]–induced monokine), CXCL10 (IFNγ‐inducible protein 10), and CXCL11 (IFN‐inducible T cell α chemoattractant) as being the most abundantly expressed chemokine family members in cutaneous LE. Expression of these ligands corresponded with the presence of a marked inflammatory infiltrate consisting of mainly CXCR3‐expressing cells, including skin‐homing lymphocytes and blood dendritic cell antigen 2–positive plasmacytoid dendritic cells (PDCs). Within cutaneous LE lesions, PDCs accumulated within the dermis and were activated to produce type I IFN, as detected by the expression of the IFNα‐inducible genes IRF7 and MxA. IFNα, in turn, was a potent and rapid inducer of CXCR3 ligands in cellular constituents of the skin. Furthermore, we demonstrated that the inflammatory CXCR3 ligands cooperate with the homeostatic chemokine CXCL12 (stromal cell–derived factor 1) during the recruitment of pathogenically relevant leukocyte subsets. Moreover, we showed that UVB irradiation induces the release of CCL27 (cutaneous T cell–attracting chemokine) from epidermal compartments into dermal compartments and up‐regulates the expression of a distinct set of chemokines in keratinocytes.

Conclusion

Taken together, our data suggest an amplification cycle in which UV light–induced injury induces apoptosis, necrosis, and chemokine production. These mechanisms, in turn, mediate the recruitment and activation of autoimmune T cells and IFNα‐producing PDCs, which subsequently release more effector cytokines, thus amplifying chemokine production and leukocyte recruitment, finally leading to the development of a cutaneous LE phenotype.
Kappa and delta opioid receptors are expressed but down‐regulated in fibroblast‐like synoviocytes of patients with rheumatoid arthritis and osteoarthritis
Authors: Shen Hua , Aeschlimann André , Reisch Nataša , Gay Renate E. , Simmen Beat R. , Michel Beat A. , Gay Steffen , Sprott Haiko
Published: May 2005

Objective

To investigate the expression and regulation of the κ‐opioid receptor (KOR) and the δ‐opioid receptor (DOR) in fibroblast‐like synoviocytes (FLS) from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), and to explore the potential antiarthritic mechanisms of peripheral KORs.

Methods

FLS isolated from synovial tissues of 6 OA patients, 8 RA patients, and 2 healthy individuals were exposed to the selective KOR agonist U69593, the selective DOR agonist SNC 80, and κ‐opioid dynorphin A in the presence or absence of the KOR antagonist nor‐binaltorphimine, the DOR antagonist naltrindole, and the proinflammatory cytokines tumor necrosis factor α (TNFα) and interleukin‐1β (IL‐1β). The expression of KOR and DOR in OA and RA FLS was evaluated on the messenger RNA (mRNA) and protein levels with TaqMan real‐time reverse transcriptase–polymerase chain reaction and immunofluorescence staining, respectively. KOR/DOR‐mediated activation of ERK‐1 and ERK‐2 was investigated by Western blotting.

Results

We detected functional KOR and DOR in normal FLS and observed a reduction of both receptors in OA and RA FLS, which was more distinct in RA FLS. U69593 enhanced KOR mRNA expression in both OA and RA FLS in a KOR antagonist–reversible manner. However, the dose required for maximal enhancement in RA FLS was 10 times higher than that required in OA FLS. TNFα and IL‐1β both suppressed the expression of DOR and KOR mRNA in both OA and RA FLS.

Conclusion

DOR and KOR are constitutively present in normal FLS and are suppressed under inflammatory conditions, such as RA and OA. Most interestingly, the KOR agonist U69593 may exert an antiarthritic effect via up‐regulation of KOR in OA and RA FLS.
Why is it difficult to find comprehensive information? Implications of information scatter for search and design
Authors: Bhavnani Suresh K.
Published: May 2005

The rapid development of Web sites providing extensive coverage of a topic, coupled with the development of powerful search engines (designed to help users find such Web sites), suggests that users can easily find comprehensive information about a topic. In domains such as consumer healthcare, finding comprehensive information about a topic is critical as it can improve a patient's judgment in making healthcare decisions, and can encourage higher compliance with treatment. However, recent studies show that despite using powerful search engines, many healthcare information seekers have difficulty finding comprehensive information even for narrow healthcare topics because the relevant information is scattered across many Web sites. To date, no studies have analyzed how facts related to a search topic are distributed across relevant Web pages and Web sites. In this study, the distribution of facts related to five common healthcare topics across high‐quality sites is analyzed, and the reasons underlying those distributions are explored. The analysis revealed the existence of few pages that had many facts, many pages that had few facts, and no single page or site that provided all the facts. While such a distribution conforms to other information‐related phenomena, a deeper analysis revealed that the distributions were caused by a trade‐off between depth and breadth, leading to the existence of general, specialized, and sparse pages. Furthermore, the results helped to make explicit the knowledge needed by searchers to find comprehensive healthcare information, and suggested the motivation to explore distribution‐conscious approaches for the development of future search systems, search interfaces, Web page designs, and training.
A proposed mechanism for intermediate atresia ani (AA), based on a porcine case of AA and hypospadias
Authors: Finnigan Daniel F. , Fisher Ken R.S. , Vrablic Olga , Halina Winn G. , Partlow Gary D.
Published: May 2005

BACKGROUND

Atresia Ani (AA) is one of the most common malformations of the lower gastrointestinal tract. It occurs in 1 in 1500 to 1 in 5000 human births. These patients suffer physically and psychologically from this disorder, and thus there is a research interest in the condition. There are 3 subcategories of AA: high, intermediate, and low. Each of these forms is likely related to separate etiological processes.

METHODS

An anatomical study was performed on a porcine case of AA with a urorectal fistula and malformed urethra.

RESULTS

This animal was found to have the intermediate form of AA.

CONCLUSIONS

A new mechanism is hypothesized, distinct from that described for the high and low forms of AA. This proposed mechanism involves the persistence of the cloacal membrane. Evidence to support this hypothesis includes: location of the urorectal fistula in the region of the embryonic cloacal duct, the lack of anomalies usually seen in conjunction with AA associated with mutations of the Sonic Hedgehog gene, and the presence of a malformed urethra, which is significantly correlated to intermediate AA. This form of AA could be related to a failure of the cloacal membrane to break down at the appropriate time during urorectal separation. Birth Defects Research (Part A), 2005. © 2005 Wiley‐Liss, Inc.
Effects of epidermal growth factor (EGF), transforming growth factor‐α (TGFα), and 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin on fusion of embryonic palates in serum‐free organ culture using wild‐type, EGF knockout, and TGFα knockout mouse strains
Authors: Abbott Barbara D. , Buckalew Angela R. , Leffler Korin E.
Published: May 2005

BACKGROUND

2,3,7,8‐Tetrachlorodibenzo‐p‐dioxin (TCDD) is teratogenic in mice, producing cleft palate (CP). TCDD exposure disrupts expression of epidermal growth factor (EGF) receptor, EGF, and transforming growth factor‐α (TGFα) in the palate and affects proliferation and differentiation of medial epithelial cells. EGF knockout embryos are less susceptible to the induction of CP by TCDD. This study used palate organ culture to examine the hypothesis that EGF enables a response to TCDD.

METHODS

The midfacial tissues from wild‐type (WT), EGF knockout, C57BL/6J, and TGFα knockout embryos were placed in organ culture on gestational day (GD) 12. Palatal explants were cultured for 4 days in serum‐free Bigger's (BGJ) medium with 0.1% dimethyl sulfoxide (DMSO) or 1 × 10−8 M TCDD with or without 2 ng of EGF/ml, 1 or 2 ng of TGFα/ml. Effects on palatal fusion were evaluated on day 4 of culture. EGF levels in explants and medium were determined using Luminex technology.

RESULTS

In serum‐free, control medium, palates from all of the strains fused. EGF knockout palates cultured with TCDD (no EGF) fused, but those cultured with TCDD + 2 ng of EGF/ml failed to fuse (p < 0.05 vs. control or TCDD without EGF). TGFα knockout palates failed to fuse when cultured with TCDD + 2 ng of TGFα/ml. EGF levels increased in tissue and accumulated in the medium after 24 hr of culture.

CONCLUSIONS

This study demonstrated that providing EGF to the palates of EGF knockout mice restored the response to TCDD. These studies support the hypothesis that the mechanism for induction of CP by TCDD is mediated via the EGFR pathway. Birth Defects Research (Part A), 2005. Published 2005 Wiley‐Liss, Inc.
Two generation reproduction study of styrene by inhalation in Crl‐CD rats
Authors: Cruzan George , Faber Willem D. , Johnson Keith A. , Roberts Linda S. , Hellwig Juergen , Carney Ed , Yarrington John T. , Stump Donald G.
Published: May 2005

This study was conducted to evaluate the potential adverse effects of styrene on reproductive capability from whole‐body inhalation exposure of F0 and F1 parental animals. Assessments included gonadal function, estrous cyclicity, mating behavior, conception rate, gestation, parturition, lactation, and weaning in the F0 and F1 generations, and F1 generation offspring growth and development. Four groups of male and female Crl:CD(SD)IGS BR rats (25/sex/group) were exposed to 0, 50, 150, and 500 ppm styrene for 6 hr daily for at least 70 consecutive days prior to mating for the F0 and F1 generations. Inhalation exposure for the F0 and F1 females continued throughout mating and gestation through gestation day 20. Inhalation exposure of the F0 and F1 females was suspended from gestation day 21 through lactation day 4. On lactation days 1 through 4, the F0 and F1 females received styrene in virgin olive oil via oral gavage at dose levels of 66, 117, and 300 mg/kg/day (divided into three equal doses, approximately 2 hr apart). These oral dosages were calculated to provide similar maternal blood peak concentrations as provided by the inhalation exposures. Inhalation exposure of the F0 and F1 females was re‐initiated on lactation day 5. Styrene exposure did not affect survival or clinical observations. Rats in the 150‐ and 500‐ppm groups in both parental generations gained weight more slowly than the controls. There were no indications of adverse effects on reproductive performance in either the F0 or F1 generation. Male and female mating and fertility indices, pre‐coital intervals, spermatogenic endpoints, reproductive organ weights, lengths of estrous cycle and gestation, live litter size and postnatal survival were similar in all exposure groups. Additionally, ovarian follicle counts and corpora lutea counts for the F1 females in the high‐exposure group were similar to the control values. No adverse exposure‐related macroscopic pathology was noted at any exposure level in the F0 and F1 generations. A previously characterized pattern of degeneration of the olfactory epithelium that lines the dorsal septum and dorsal and medial aspects of the nasal turbinates occurred in the F0 and F1 generation animals from the 500‐ppm group. In the 500‐ppm group, F2 birthweights were reduced compared to the control and F2 offspring from both the 150‐ and 500‐ppm exposure groups gained weight more slowly than the controls. Based on the results of this study, an exposure level of 50 ppm was considered to be the NOAEL for F0 and F1 parental systemic toxicity; the NOAEL for F0 and F1 reproductive toxicity was 500 ppm or greater. Birth Defects Res B 2005. © 2005 Wiley‐Liss, Inc.
Antimicrobial peptides: New candidates in the fight against bacterial infections
Authors: Toke Orsolya
Published: May 2005

Antimicrobial peptides (AMPs) of innate origin are agents of the most ancient form of defense systems. They can be found in a wide variety of species ranging from bacteria through insects to humans. Through the course of evolution, host organisms developed arsenals of AMPs that protect them against a large variety of invading pathogens including both Gram‐negative and Gram‐positive bacteria. At a time of increasing bacterial resistance, AMPs have been the focus of investigation in a number of laboratories worldwide. Although recent studies show that some of the peptides are likely to have intracellular targets, the vast majority of AMPs appear to act by permeabilization of the bacterial cell membrane. Their activity and selectivity are governed by the physicochemical parameters of the peptide chains as well as the properties of the membrane system itself. In this review, we will summarize some of the recent developments that provide us with a better understanding of the mode of action of this unique family of antibacterial agents. Particular attention will be given to the determinants of AMP–lipid bilayer interactions as well as to the different pore formation mechanisms. The emphasis will be on linear AMPs but representatives of cysteine‐bridged AMPs will also be discussed. © 2005 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 80: 717–735, 2005

This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com
“Parallel” and “Antiparallel Tail‐Clamps” Increase the Efficiency of Triplex Formation with Structured DNA and RNA Targets
Authors: Nadal Anna , Eritja Ramon , Esteve Teresa , Pla Maria
Published: May 2005

Sequence‐specific triple‐helix structures can be formed by parallel and antiparallel DNA clamps interacting with single‐stranded DNA or RNA targets. Single‐stranded nucleic acid molecules are known to adopt secondary structures that might interfere with intermolecular interactions. We demonstrate the correlation between a secondary structure involving the target—a stable stem predicted by in silico folding and experimentally confirmed by thermal stability and competition analyses—and an inhibitory effect on triplex formation. We overcame structural impediments by designing a new type of clamp: “tail‐clamps”. A combination of gel‐shift, kinetic analysis, UV thermal melting and thermodynamic techniques was used to demonstrate that tail‐clamps efficiently form triple helices with a structured target sequence. The performance of parallel and antiparallel tail‐clamps was compared: antiparallel tail‐clamps had higher binding efficiencies than parallel tail‐clamps both with structured DNA and RNA targets. In addition, the reported triplex‐stabilizing property of 8‐aminopurine residues was confirmed for tail‐clamps. Finally, we discuss the possible use of this improved triplex technology as a new tool for applications in molecular biology.
Chemical Synthesis of Triple‐Labelled Three‐Helix Bundle Binding Proteins for Specific Fluorescent Detection of Unlabelled Protein
Authors: Engfeldt Torun , Renberg Björn , Brumer Harry , Nygren Per Åke , Eriksson Karlström Amelie
Published: May 2005

Site‐specifically triple‐labelled three‐helix bundle affinity proteins (affibody molecules) have been produced by total chemical synthesis. The 58 aa affinity proteins were assembled on an automated peptide synthesizer, followed by manual on‐resin incorporation of three different reporter groups. An orthogonal protection strategy was developed for the site‐specific introduction of 5‐(2‐aminethylamino)‐1‐naphthalenesulfonic acid (EDANS) and 6‐(7‐nitrobenzofurazan‐4‐ylamino)‐hexanoic acid (NBDX), constituting a donor/acceptor pair for fluorescence resonance energy transfer (FRET), and a biotin moiety, used for surface immobilization. Circular dichroism and biosensor studies of the synthetic proteins and their recombinant counterparts revealed that the synthetic proteins were folded and retained their binding specificities. The biotin‐conjugated protein could be immobilized onto a streptavidin surface without loss of activity. The synthetic, doubly fluorescent‐labelled affinity proteins were shown to function as fluorescent biosensors in an assay for the specific detection of unlabelled human IgG and IgA.
Design and Synthesis of Hierarchical Materials from Ordered Zeolitic Building Units
Authors: Kirschhock Christine E. A. , Kremer Sebastien P. B. , Vermant Jan , Van Tendeloo Gustaaf , Jacobs Pierre A. , Martens Johan A.
Published: May 2005

The crystallization of colloidal silicalite‐1 from clear solution is one of the best understood zeolite formation processes. Colloidal silicalite‐1 formation involves a self‐assembly process in which nanoslabs and nanotablets with a silicalite‐1 type connectivity are formed at intermediate stages. During the assembly process, with strongly anisometric particles present, regions appear with orientational correlations, as evidenced with measurements of dynamic light scattering, viscosity, and rotation of polarized light. The presence of such regions rationalizes the unexpected differences between the crystallization kinetics under microgravity and on earth. The discovery of the locally oriented regions sheds new light on currently poorly understood hydrodynamic effects on the zeolite formation processes, such as the influence of stirring on the phases obtained and the subsequent kinetics. Addition of surfactants or polymers modifies the ordering of the zeolitic building units in the correlated regions, and new types of hierarchical materials named zeogrids and zeotiles can be obtained.
Evidence that one subset of anaplastic thyroid carcinomas are derived from papillary carcinomas due to BRAF and p53 mutations
Authors: Quiros Roderick M. , Ding Helen G. , Gattuso Paolo , Prinz Richard A. , Xu Xiulong
Published: May 2005

BACKGROUND

Anaplastic thyroid carcinoma (ATC) is the most lethal form of thyroid neoplasia and represents the end stage of thyroid tumor progression. In the current study, genetic alterations in a panel of ATC were profiled to determine the origins of ATC.

METHODS

Eight ATC were analyzed for BRAF mutation at codon 599 by using mutant‐allele‐specific polymerase chain reaction (PCR) and DNA sequencing of the PCR‐amplified exon 15. RAS mutation (HRAS, KRAS, and NRAS) at codons 12, 13, and 61 was analyzed by direct sequencing of PCR‐amplified exons 1 and 2 of the RAS gene. RET/PTC rearrangements and p53 mutation were monitored by immunohistochemical (IHC) staining by anti‐RET antibodies and an anti‐p53 mAb, respectively.

RESULTS

BRAF was mutated in 5 of the 8 ATCs tested. Histologic examination revealed that 4 of these 5 BRAF‐mutated ATCs contained a PTC component, suggesting that they may be derived from BRAF‐mutated PTC. Of the 3 ATCs with wild‐type BRAF, 2 had spindle cell features; one had follicular neoplastic characteristics mixed with papillary structures. Analysis of RAS mutation revealed only an HRAS mutation at codon 11, due to the transversion of GCC to TCC in one ATC with wild‐type BRAF. This leads to the substitution of valine to serine. IHC analysis of RET/PTC rearrangements revealed no positive staining of RET in any of 8 ATCs, suggesting that these ATCs are not derived from RET/PTC‐ rearranged PTC. In contrast, IHC analysis of p53 mutation revealed that p53 was detected in the nuclei of 5 of 5 BRAF‐mutated ATCs and 2 of 3 ATCs with wild‐type BRAF. p53 staining was present only in anaplastic thyroid tumor cells but not in neighboring papillary thyroid tumor cells.

CONCLUSIONS

These results suggest that many ATCs with papillary components are derived from BRAF‐mutated PTC, because of the addition of p53 mutation. Cancer 2005. © 2005 American Cancer Society.
Epidermal growth factor receptor‐targeted treatment for advanced colorectal carcinoma
Authors: Venook Alan P.
Published: May 2005

Substantial effort has focused on the development of novel targeted agents for treating patients with late‐stage colorectal carcinoma. These agents are designed specifically to inhibit biochemical processes associated with pathogenesis. Numerous molecules targeting the epidermal growth factor receptor have been investigated as therapeutic agents and appear to herald a shift in the treatment paradigm for colorectal carcinoma. Cancer 2005. © 2005 American Cancer Society.
Colorectal carcinoma screening among Hispanics and non‐Hispanic whites in a rural setting
Authors: Thompson Beti , Coronado Gloria , Neuhouser Marian , Chen Lu
Published: May 2005

BACKGROUND

Colorectal carcinoma ranks as the second most common cancer and the second leading cause of cancer death in the United States. Hispanics are less likely than their non‐Hispanic white counterparts to have ever received a fecal occult blood test (FOBT) or sigmoidoscopy/colonoscopy. Little is known about the barriers to screening in the Hispanic population.

METHODS

The authors used baseline data from a community randomized trial of cancer prevention to compare screening prevalence and the associations between reported barriers and screening participation between Hispanics (n = 137) and non‐Hispanic whites (n = 491) age ≥ 50 years.

RESULTS

Hispanics were less likely than non‐Hispanic whites to have ever received an FOBT (P = 0.003) or sigmoidoscopy/colonoscopy (P = 0.001). No significant difference across ethnic groups was observed in the prevalence of recent screening using FOBT (29.8% for Hispanics vs. 34.5% for non‐Hispanic whites; P = 0.41), but recent use of sigmoidoscopy/colonoscopy was lower for Hispanics (24.1% for Hispanics vs. 33.7% for non‐Hispanic whites; P 0.06). Lacking health care coverage or having few years of education were directly associated with failure to ever receive an FOBT or sigmoidoscopy/colonoscopy.

CONCLUSIONS

Interventions to improve adherence to colorectal carcinoma screening recommendations among Hispanics should target initial screening examinations, particularly among those lacking health care coverage or having low levels of education. Cancer 2005. © 2005 American Cancer Society.
Experimental analysis of the transdifferentiation of visceral to parietal endoderm in the mouse
Authors: Ninomiya Youichirou , Davies Timothy J. , Gardner Richard L.
Published: May 2005

The visceral endoderm (VE) of isolated extraembryonic regions (ExEmbs) of 7 days postcoitum (dpc) prestreak mouse conceptuses have been shown to convert readily to parietal endoderm (PE). The present study addresses the following three unanswered questions. On what does conversion depend, how rapidly does it occur, and is it an enduring general property of a residual small population of relatively immature cells? In situ hybridization reveals that change in cell state occurs within 2 days of culture. Deprivation of the mesoderm also promotes it in later ExEmbs. Conversely, the conversion to PE in isolated 7 dpc ExEmbs is suppressed by grafting 8 dpc or 9 dpc mesoderm. Hence, the conversion provides an example of transdifferentiation that is promoted by the absence of extraembryonic mesoderm. The presence of mesoderm seems to be necessary to enable the VE to grow rather than convert to PE, as occurs if it retains contact with the extraembryonic ectoderm. Developmental Dynamics 233:837–846, 2005. © 2005 Wiley‐Liss, Inc.
Reduced expression of the PTH/PTHrP receptor during development of the mammary gland influences the function of the nipple during lactation
Authors: Kobayashi Tatsuya , Kronenberg Henry M. , Foley John
Published: May 2005

Signaling by the parathyroid hormone/parathyroid hormone‐related protein receptor (Ppr) is necessary for mammary gland development beyond the early induction stage in mice. We used a series of murine models of reduced Ppr expression to determine how diminished receptor signaling influences mammary development. Reduction of Ppr expression to very low levels prevented mammary gland development. A less‐severe reduction in Ppr expression permitted progression of mammary gland development beyond the induction stage, but the nipples of these mice were dramatically smaller than those of controls, with altered epidermis and connective tissue. Mothers with reduced expression of Ppr could not successfully nurse pups; however, the lactating glands did produce milk but could not efficiently deliver it. This finding was associated with reduced levels of matrix metalloproteinase‐2 and an absence of pregnancy‐associated remodeling of connective tissue matrix in the nipple. Reduced smooth muscle appears to underlie the majority of nipple deficiencies in mice with lower levels of the Ppr expression. Developmental Dynamics 233:794–803, 2005. © 2005 Wiley‐Liss, Inc.
Expression of all Wnt genes and their secreted antagonists during mouse blastocyst and postimplantation development
Authors: Kemp Caroline , Willems Erik , Abdo Shaaban , Lambiv Louis , Leyns Luc
Published: May 2005

In this extensive study, real‐time reverse transcriptase‐polymerase chain reaction was used to analyze the expression levels of all 19 Wnt genes and their 11 potential antagonists in mouse blastocysts, pregastrula, gastrula, and neurula stages. By complementing these results with in situ hybridization, we revealed new expression domains for Wnt2b and Sfrp1, respectively, in the future primitive streak at the posterior side and in the anterior visceral endoderm before the initiation of gastrulation. Moreover, the anterior visceral endoderm expresses three secreted Wnt antagonists (Sfrp1, Sfrp5, and Dkk1) in partially overlapping domains. We also identified expression patterns for the Wnt1, Wnt3a, Wnt6, Wnt7b, Wnt9a, Wnt10b, and Sfrp1 genes at the blastocyst stage. In particular, the expression of Wnt1 and Sfrp1 predominantly in the inner cell mass and of Wnt9a in the mural trophoblast and inner cell mass cells surrounding the blastocoele suggests new roles for the Wnt pathway in preimplantation development. This article is the first report on the regional expression of Wnt genes in the mouse blastocyst. Developmental Dynamics 233:1064–1075, 2005. © 2005 Wiley‐Liss, Inc.
Genotoxicity and physicochemical characteristics of traffic‐related ambient particulate matter
Authors: de Kok Theo M. , Hogervorst Janneke G. , Briedé Jacco J. , van Herwijnen Marcel H. , Maas Lou M. , Moonen Edwin J. , Driece Hermen A. , Kleinjans Jos C.
Published: May 2005

Exposure to ambient particulate matter (PM) has been linked to several adverse health effects. Since vehicular traffic is a PM source of growing importance, we sampled total suspended particulate (TSP), PM10, and PM2.5 at six urban locations with pronounced differences in traffic intensity. The mutagenicity, DNA‐adduct formation, and induction of oxidative DNA damage by the samples were studied as genotoxicological parameters, in relation to polycyclic aromatic hydrocarbon (PAH) levels, elemental composition, and radical‐generating capacity (RGC) as chemical characteristics. We found pronounced differences in the genotoxicity and chemical characteristics of PM from the various locations, although we could not establish a correlation between traffic intensity and any of these characteristics for any of the PM size fractions. Therefore, the differences between locations may be due to local sources of PM, other than traffic. The concentration of total (carcinogenic) PAHs correlated positively with RGC, direct and S9‐mediated mutagenicity, as well as the induction of DNA adducts and oxidative DNA damage. The interaction between total PAHs and transition metals correlated positively with DNA‐adduct formation, particularly from the PM2.5 fraction. RGC was not associated with one specific PM size fraction, but mutagenicity and DNA reactivity after metabolic activation were relatively high in PM10 and PM2.5, when compared with TSP. We conclude that the toxicological characteristics of urban PM samples show pronounced differences, even when PM concentrations at the sample sites are comparable. This implies that emission reduction strategies that take chemical and toxicological characteristics of PM into account may be useful for reducing the health risks associated with PM exposure. Environ. Mol. Mutagen., 2005. © 2005 Wiley‐Liss, Inc.
Antimutagenicity of green tea polyphenols in the liver of transgenic medaka
Authors: Winn Richard N. , Kling Heather , Norris Michelle B.
Published: May 2005

We examined the ability of a mixture of the predominant green tea polyphenolic compounds (GTP) to reduce benzo[a]pyrene (B[a]P)‐induced mutations in the cII gene of the λ transgenic medaka. Fish were treated with 50 ppb B[a]P for 24 hr, followed by exposure to 2 ppm or 10 ppm GTP for 28 days. cII mutations in livers of fish exposed to B[a]P were increased significantly, 2.6‐fold above controls. In contrast, the addition of GTP significantly reduced the frequency of cII mutants by 84%, comparable to that of controls. The frequencies of mutations at G:C basepairs, mutations that are highly characteristic of B[a]P exposure, were elevated significantly in treated fish. By comparison, B[a]P‐exposed fish also treated with GTP showed reductions in these mutations, demonstrating a protective effect of GTP against B[a]P‐induced mutagenesis. The antioxidant mechanism of GTP possibly played an important role in the reduction of B[a]P mutagenicity. These results corroborate findings from rodent models, showing that the protective effects of green tea extend to different species, and suggesting that similar mechanisms of B[a]P mutagenesis and GTP antimutagenesis are shared among the models. These studies illustrate the utility of λ transgenic medaka for in vivo mutation analyses and suggest that this fish may be a valuable model in chemoprevention studies. Environ. Mol. Mutagen., 2005. © 2005 Wiley‐Liss, Inc.
Further development of the β‐lactamase MutaGen assay and evaluation by comparison with Ames fluctuation tests and the umu test
Authors: Reifferscheid Georg , Arndt Christian , Schmid Claudia
Published: May 2005

A rapid, high‐throughput bacterial mutagenicity test system has been developed (MutaGen test) that detects reversions of inactivating base‐pair substitutions and frameshifts in a TEM‐1 class A β‐lactamase (ampicillinase) gene. To quickly and sensitively detect mutagens, the system utilises a series of plasmids that contain the mutated ampicillinase gene and the mucAB operon. Inactivating mutations in the ampicillinase gene include frameshifts integrated into repetitive GC‐sequences and G‐runs known to be mutagenic hot‐spots, and base‐pair substitutions inserted in or around the β‐lactamase active site. Frameshift mutations completely inactivated the enzyme only when located downstream of the active‐site serine (Ser68). Previous (reporter gene based) assays with this system have detected reversion to ampicillin resistance by luminescence driven by induction of the tet‐promotor controlled lacZ gene. In the present study, we describe the construction and evaluation of 19 additional potential tester strains. We also developed conditions for detecting reversions by pH shift using bromocresol purple and by directly detecting the enzymatic activity of β‐lactamase using nitrocefin. A 384‐well format version of the pH shift MutaGen test was used to assay more than 20 chemicals. The responses in the assay were compared with responses for the same chemicals in the umu test and Ames fluctuation assays. The results indicate that the MutaGen test has high specificity for detecting specific mutations and, in some instances, better sensitivity than the other tests. Since the test is easy to conduct, sterile working conditions are not necessary, and the mutagenicity results are available either within one working day or overnight, the assay shows promise for the rapid screening of potentially genotoxic substances. Environ. Mol. Mutagen., 2005. © 2005 Wiley‐Liss, Inc.
Transgenic mouse lines expressing synaptopHluorin in hippocampus and cerebellar cortex
Authors: Araki Rikita , Sakagami Hiroyuki , Yanagawa Yuchio , Hikima Takuya , Ishizuka Toru , Yawo Hiromu
Published: May 2005

We generated six transgenic mouse lines in which synaptopHluorin (SpH), one of green fluorescent protein‐based sensors of vesicular exocytosis, was expressed under the control of neuron‐specific Thy‐1.2 promoter. In situ hybridization study revealed that SpH mRNA was expressed in a broad spectrum of brain regions in four of them, whereas in others it was expressed in the specific regions of the hippocampus. In one particular line, SpH immunoreactivity was specifically observed in the mossy fiber presynaptic terminals of both hippocampus and cerebellar cortex. The fluorescence intensity of these presynaptic terminals was somewhat decreased by acidic buffer superfusion and greatly increased by vesicular neutralization of pH, indicating that the SpH molecules are mainly distributed in the synaptic vesicles. The exocytosis‐dependent fluorescence increment was measured upon activation of a single presynaptic terminal. These transgenic lines are expected to facilitate morphological and physiological studies of presynaptic terminals in a variety of regions of the brain. genesis 42:53–60, 2005. © 2005 Wiley‐Liss, Inc.
Mood stabilization in the treatment of bipolar disorder: focus on quetiapine
Authors: Vieta Eduard
Published: May 2005

An Erratum has been published for this article in Human Psychopharmacology 20(5), 2005, 375.

The use of at least one mood‐stabilizing agent is common clinical practice in the treatment of bipolar disorder, regardless of the treatment setting or disease phase. However, a consensus definition of ‘mood stabilizer’ remains to be established. A mood stabilizer has been operationally described as an agent that is useful in at least one phase of bipolar disorder while not worsening any other phase of the illness. More stringent definitions have been proposed, and it can be argued that a clinically effective mood stabilizer would have efficacy in a broad range of affective, psychotic, behavioral and cognitive domains in all phases of bipolar disorder and would be well tolerated across a range of doses for sustained periods. Clinically effective mood stabilizers should treat mania and depression, while preventing recurrence and improving quality of life. Effective treatment should not precipitate mania, depression, or rapid cycling, and should minimize the burden of treatment‐emergent side effects. Data from clinical studies of quetiapine are reviewed in context with the literature discussing traditional and emerging mood stabilizers. Using a liberal definition, the evidence for quetiapine qualifies it as a bimodal mood stabilizer based on its demonstrated effectiveness in the treatment of bipolar mania and depression. Further data suggest that quetiapine has promise across all phases of bipolar disorder with the potential to meet even the most stringent definitions of a mood stabilizer. Copyright © 2005 John Wiley & Sons, Ltd.
Changes in mechanical properties and cellularity during long‐term culture of collagen fiber ACL reconstruction scaffolds
Authors: Caruso Andrea B. , Dunn Michael G.
Published: May 2005

Resorbable scaffolds for anterior cruciate ligament (ACL) reconstruction should provide temporary mechanical function then gradually breakdown while promoting matrix synthesis by local cells. Crosslinking influences collagen's mechanical properties, degradation rate, and interactions with cells. Our objective was to compare the effects of different crosslinkers on cellularity and mechanical properties during long‐term (8 week) culture of collagen fiber scaffolds. Fibers were fabricated from an acid‐insoluble dispersion of bovine dermal collagen and crosslinked with either ultraviolet irradiation (UV; a physical crosslinker) or 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide (EDC; a chemical crosslinker). Scaffolds consisted of 50 fibers bundled in parallel. Initial attachment of fibroblasts was similar on both scaffolds; however, from 1 to 8 weeks in culture, UV‐crosslinked scaffolds had significantly more cells attached than EDC‐crosslinked scaffolds. The initial breaking load (3.50 N) and stiffness (2.23 N/mm) of EDC‐crosslinked scaffolds were significantly greater than those of UV‐crosslinked scaffolds (2.32 N; 1.21 N/mm) and were unaffected by long‐term fibroblast culture. In contrast, the load‐bearing capacity of fibroblast‐seeded UV‐crosslinked scaffolds decreased 60% to 0.91 N after 8 weeks in culture. EDC‐crosslinked scaffolds maintained strength and moderate cellularity; UV‐crosslinked scaffolds, in contrast, were highly cellular, but had poor mechanical properties that decreased during culture. These in vitro results suggest that collagen fiber scaffolds crosslinked with EDC may be more suitable for ACL reconstruction than those crosslinked with UV. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
Development of a resorbable macroporous cellulosic material used as hemostatic in an osseous environment
Authors: Laurence S. , Bareille R. , Baquey C. , Fricain J.C.
Published: May 2005

The control of bleeding is a frequently encountered therapeutic problem, particularly during dental surgery. The most efficient substances used to resolve this problem are not risk‐free because of their animal or human origins, so cellulosic materials are potentially of interest. The aim of this study was to develop a resorbable macroporous cellulosic material for use as a resorbable hemostatic agent in bone sites. The degradation and the cytocompatibility of the cellulosic material versus controls were evaluated and its behaviour in vivo was studied. An original process using calcium carbonate powder as inverse matrix was used to develop a macroporous material. In order to predegrade the cellulosic material for hemostatic use, oxidation was performed with periodate. A dialdehyde component unstable at physiological pH was thus obtained. The material was found to have cytotoxicity, biocompatibility, and resorption properties similar to control but its hemostatic power was higher. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
The Italian Registry for Therapeutic Apheresis. A report from the Apheresis Study Group of the Italian Society of Nephrology
Authors: Passalacqua Stefano , Staffolani Emiliano , Busnach Ghil , Roccatello Dario , Pasquali Sonia , Cappelli Paolo , Liuzzo Gabriele
Published: May 2005

Many clinical indications and different technical issues have been reported on therapeutic apheresis: much criticism has also been recorded in several instances, mainly due to the lack of large clinical trials to validate collected data. A Registry where all the available data can be organized and analyzed therefore becomes a priority for all the professionals involved in apheresis. The purpose of this report is to describe the data submitted from 1994 to 2004 from 15,285 treatments on 1,477 patients from 44 Centers, including mainly, but not exclusively, Nephrological Units, collected by the Apheresis Study Group of the Italian Society of Nephrology in 15 Italian regions. Plasma exchange accounted for 56.2% of the procedures, and of these 50.4% were performed by filtration. Plasma treatment was used in 40.1% of procedures, namely with Protein A immunoadsorption (14.6%), LDL‐Cholesterol dextran sulfate adsorption (9.7%), and semiselective cascade or double filtration (12.6%). Cell apheresis, limited to photopheresis, was used in 0.85% of cases, and whole blood treatment (direct adsorption lipoprotein, and molecular adsorption recirculating system) in 2.7%. The procedures analyzed here account for less than 20% of estimated therapeutic apheresis performed in Italy, according to the national survey of activity performed for year 2000 by the Italian Apheresis Society. Notwithstanding that the data are largely incomplete, they are sufficiently informative for a definite trend: plasma treatment with filtration on fractionation filters and adsorption must be used as often as possible, instead of plasma exchange, thus obtaining the most selective removals. J. Clin. Apheresis © 2005 Wiley‐Liss, Inc.
Cellular and molecular interactions regulating skeletogenesis
Authors: Colnot Céline
Published: May 2005

Skeletal development involves complex coordination among multiple cell types and tissues. In long bones, a cartilage template surrounded by the perichondrium is first laid down and is subsequently replaced by bone marrow and bone, during a process named endochondral ossification. Cells in the cartilage template and the surrounding perichondrium are derived from mesenchymal cells, which condense locally. In contrast, many cell types that make up mature bone and in particular the bone marrow are brought in by the vasculature. Three tissues appear to be the main players in the initiation of endochondral ossification: the cartilage, the adjacent perichondrium, and the invading vasculature. Interactions among these tissues are synchronized by a large number of secreted and intracellular factors, many of which have been identified in the past 10 years. Some of these factors primarily control cartilage differentiation, while others regulate bone formation and/or angiogenesis. Understanding how these factors operate during skeletal development through the analyses of genetically altered mice depends on being able to distinguish the effect of these molecules on the different cell types that comprise the skeleton. This review will discuss the complexity of skeletal phenotypes, which arises from the tightly regulated, complex interactions among the three tissues involved in bone development. Specific examples illustrate how gene functions may be further assessed using new approaches including genetic and tissue manipulations. © 2005 Wiley‐Liss, Inc.
Effect of adenosine triphosphate on phosphate uptake in renal proximal tubule cells: Involvement of PKC and p38 MAPK
Authors: Lee Yun Jung , Park Soo Hyun , Jeung Tae Oh , Kim Kee Won , Lee Jang Hern , Han Ho Jae
Published: May 2005

ATP has been known to act as an extracellular signal and to be involved in various functions of kidney. Renal proximal tubular reabsorption of phosphate (Pi) contributes to the maintenance of phosphate homeostasis, which is regulated by Na+/Pi cotransporter. However, the effects of ATP on Na+/Pi cotransporters were not elucidated in proximal tubule cells (PTCs). Thus, the effects of ATP on Na+/Pi cotransporter and its related signal pathways are examined in the primary cultured renal PTCs. In the present study, ATP inhibited Pi uptake in a time (> 1 h) and dose (>10−6M) dependent manner. ATP‐induced inhibition of Pi uptake was correlated with the decrease of type II Na+/Pi cotransporter mRNA. ATP‐induced inhibition of Pi uptake may be mediated by P2Y receptor activation, since suramin (non‐specific P2 receptor antagonist) and RB‐2 (P2Y receptor antagonist) blocked it. ATP‐induced inhibition of Pi uptake was blocked by neomycin, U73122 (phospholipase C (PLC) inhibitors), bisindolylmaleimide I, H‐7, and staurosporine (protein kinase C (PKC) inhibitors), suggesting the role of PLC/PKC pathway. ATP also increased inositol phosphates (IPs) formation and induced PKC translocation from cytosolic fraction to membrane fraction. In addition, ATP‐induced inhibition of Pi uptake was blocked by SB 203580 [a p38 mitogen activated protein kinase (MAPK) inhibitor], but not by PD 98059 (a p44/42 MAPK inhibitor). Indeed, ATP induced phosphorylation of p38 MAPK, which was not blocked by PKC inhibitor. In conclusion, ATP inhibited Pi uptake via PLC/PKC as well as p38 MAPK in renal PTCs. © 2005 Wiley‐Liss, Inc.
Disruption of Sertoli‐germ cell adhesion function in the seminiferous epithelium of the rat testis can be limited to adherens junctions without affecting the blood–testis barrier integrity: An in vivo study using an androgen suppression model
Authors: Xia Weiliang , Wong Ching Hang , Lee Nikki P.Y. , Lee Will M. , Cheng C. Yan
Published: May 2005

During spermatogenesis, both adherens junctions (AJ) (such as ectoplasmic specialization (ES), a testis‐specific AJ type at the Sertoli cell‐spermatid interface (apical ES) or Sertoli–Sertoli cell interface (basal ES) in the apical compartment and BTB, respectively) and tight junctions (TJ) undergo extensive restructuring to permit germ cells to move across the blood–testis barrier (BTB) as well as the seminiferous epithelium from the basal compartment to the luminal edge to permit fully developed spermatids (spermatozoa) to be sloughed at spermiation. However, the integrity of the BTB cannot be compromised throughout spermatogenesis so that postmeiotic germ cell‐specific antigens can be sequestered from the systemic circulation at all times. We thus hypothesize that AJ disruption in the seminiferous epithelium unlike other epithelia, can occur without compromising the BTB‐barrier, even though these junctions, namely TJ and basal ES, co‐exist side‐by‐side in the BTB. Using an intratesticular androgen suppression‐induced germ cell loss model, we have shown that the disruption of AJs indeed was limited to the Sertoli‐germ cell interface without perturbing the BTB. The testis apparently is using a unique physiological mechanism to induce the production of both TJ‐ and AJ‐integral membrane proteins and their associated adaptors to maintain BTB integrity yet permitting a transient loss of cell adhesion function by dissociating N‐cadherin from β‐catenin at the apical and basal ES. The enhanced production of TJ proteins, such as occludin and ZO‐1, at the BTB site can supersede the transient loss of cadherin‐catenin function at the basal ES. This thus allows germ cell depletion from the epithelium without compromising BTB integrity. It is plausible that the testis is using this novel mechanism to facilitate the movement of preleptotene and leptotene spermatocytes across the BTB at late stage VIII through early stage IX of the epithelial cycle in the rat while maintaining the BTB immunological barrier function. © 2005 Wiley‐Liss, Inc.