bluuebunny/crossref_metadata_2025 huggingface dataset:
Donepezil augmentation of clozapine monotherapy in schizophrenia patients: a double blind cross‐over study
Authors: Stryjer Rafael , Strous Rael , Bar Faina , Shaked Ginette , Shiloh Roni , Rozencwaig Silvio , Grupper Daniel , Buchman Nili , Kotler Moshe , Rabey J. Martin , Weizman Abraham
Published: May 2004

Increasing evidence suggests that the cholinergic system is involved in the pathogenesis of schizophrenia. Donepezil, a central cholinesterase inhibitor, improves psychotic symptomatology in demented patients, however, evidence for its role in the management of active psychosis in schizophrenia remains limited. An 18‐week double blind cross‐over study was conducted in which eight patients were randomly assigned to either donepezil (5 mg/day for the first 4 weeks and 10 mg/day for the following 4 weeks) or placebo as augmentation treatment to clozapine. After this initial phase, there was a 2‐week washout period of the study medication after which the same regimen was crossed over at the same dose and for the same period (8 weeks). No significant difference was noted in the total positive and negative symptom scale scores when donepezil was compared with placebo (16.7%+12.97% vs 3.20%+13.94% respectively, p = 0.18). However, three patients improved (>15%) in the total PANSS scores (37.03%, 16.6% and 25.33%) during the donepezil treatment phase, while only one patient improved (20.87%) during the placebo phase. No differences were noted in the Calgary depression scale (p = 0.305), Simpson Angus scale (p = 0.374), clinical global impression‐improvement scale (p = 0.23) and clinical global impression‐severity of illness scores (p = 0.116). Although this preliminary study failed to demonstrate a clear effect of donepezil augmentation in clozapine treated chronic schizophrenia patients, it seems that the subtle positive effect of donepezil observed in some of our patients should encourage further investigation in a larger sample of this patient subpopulation. Copyright © 2004 John Wiley & Sons, Ltd.
Immunohistochemical patterns in rectal cancer: Application of tissue microarray with prognostic correlations
Authors: Fernebro Eva , Bendahl Pär‐Ola , Dictor Michael , Persson Annette , Fernö Mårten , Nilbert Mef
Published: May 2004

We utilized the high‐throughput tissue microarray method to characterize immunohistochemical expression patterns with correlations to prognosis in rectal cancer. Immunostaining for the markers Ki‐67, Bcl‐2, p53, EGFR, E‐cadherin, β‐catenin, MLH1 and MSH2 was performed in 269 rectal cancers. Expression profiles were correlated to metastasis‐free survival. Immunostaining revealed frequent upregulation and/or aberrant staining patterns for several of the markers, but Ki‐67, p53, Bcl‐2 and EGFR did not show any correlation to prognosis. However, reduced membranous staining for β‐catenin (p = 0.04), lack of cytoplasmic staining for β‐catenin (p = 0.04), reduced membranous staining for E‐cadherin (p = 0.02) and lack of cytoplasmic staining for E‐cadherin (p = 0.02) correlated with metastatic disease. Multivariate analysis including the factors Dukes' stage and tumor differentiation grade demonstrated increased risk of metastatic disease in tumors with lack of cytoplasmic staining for β‐catenin (HR = 3.1, p = 0.02), reduced membranous staining for β‐catenin (HR = 1.7, p = 0.06) and reduced membranous staining for E‐cadherin (HR = 2.1, p = 0.06). Loss of MMR protein expression was confirmed to be a rare event in rectal cancer with loss of MLH1 staining in 3% and MSH2 in 1% of the tumors. The lack of prognostic information contributed by most of these markers suggests that single markers for prognosis may be of limited value in rectal cancer. However, altered expression of β‐catenin and E‐cadherin correlated with metastatic disease, and these markers may have prognostic importance in rectal cancer. © 2004 Wiley‐Liss, Inc.
Comparison of 10 serum bone turnover markers in prostate carcinoma patients with bone metastatic spread: Diagnostic and prognostic implications
Authors: Jung Klaus , Lein Michael , Stephan Carsten , Von Hösslin Katharina , Semjonow Axel , Sinha Pranav , Loening Stefan A. , Schnorr Dietmar
Published: May 2004

Our aim was to assess the diagnostic accuracy of bone markers in serum of patients with prostate cancer (PCa) for early detection of bone metastases and their usefulness as predictors of PCa‐caused mortality. In sera of 117 PCa patients (pN0M0, n = 39; pN1M0, n = 34; M1, n = 44), 35 healthy men and 35 patients with benign prostatic hyperplasia, bone formation markers [total and bone‐specific alkaline phosphatase (tALP, bALP), amino‐terminal procollagen propeptides of type I collagen (P1NP), osteocalcin (OC)], bone resorption markers [bone sialoprotein (BSP), cross‐linked C‐terminal (CTX) and cross‐linked N‐terminal (NTX) telopeptides of type I collagen, tartrate‐resistant acid phosphatase isoenzyme 5b (TRAP)] and osteoclastogenesis markers [osteoprotegerin (OPG), receptor activator of nuclear factor κB ligand (RANKL)] were measured. tALP, bALP, BSP, P1NP, TRAP, NTX and OPG were significantly increased in PCa patients with bone metastases compared to patients without metastases. OPG showed the best discriminatory power to differentiate between these patients. Logistic regression analysis resulted in a model with OPG and TRAP as variables that predicted bone metastasis with an overall correct classification of 93%. Patients with concentrations of OPG, P1NP, tALP, bALP, BSP, NTX, TRAP and CTX above cut‐off levels showed significantly shorter survival than patients with low marker concentrations. Multivariate Cox proportional hazards regression revealed that only OPG and BSP were independent prognostic factors for PCa‐related death. Thus, the importance of serum OPG in detecting bone metastatic spread, alone or in combination with other bone markers, and predicting survival in PCa patients has been clearly demonstrated. © 2004 Wiley‐Liss, Inc.
Neoexpression of N‐cadherin in E‐cadherin positive colon cancers
Authors: Rosivatz Erika , Becker Ingrid , Bamba Masamichi , Schott Christina , Diebold Joachim , Mayr Doris , Höfler Heinz , Becker Karl‐Friedrich
Published: May 2004

In our study, we aimed to investigate the expression of N‐cadherin and E‐cadherin and their dependency on epithelial‐mesenchymal transition regulators SNAI1, SIP1 and TWIST in human colon cancer. Expression of E‐cadherin and N‐cadherin was examined by immunohistochemistry in 80 colon carcinomas by using paraffin embedded and formalin fixed tissues. Those cases were partly analyzed for mRNA expression of N‐cadherin (42 cases), TWIST (18 cases), SNAI1 (25 cases) and SIP1 (25 cases) by real‐time quantitative RT‐PCR. Additionally, colon carcinomas that showed amplification of 20q13, the localization of the human SNAI1 gene, were examined. We found cytoplasmic and/or membrane‐associated immunoreactivity of N‐cadherin in 35/80 (44%) of the cases. However, there was no correlation to upregulated TWIST mRNA levels, as we have shown previously for diffuse‐type gastric cancers with abnormal N‐cadherin expression. Reduced and/or cytoplasmic E‐cadherin immunoreactivity was detected in 19% (15/80) of the cases. Expression of SNAI1 or SIP1 mRNA was not seen in any of the 25 cases analyzed. There was no correlation between amplification of 20q13 and SNAI1 mRNA expression. Remarkably, N‐cadherin was almost exclusively expressed in those cases showing normal E‐cadherin immunoreactivity, suggesting a mutual exclusion between abnormal E‐cadherin reduction and upregulation of N‐cadherin. For the first time, we postulate a role for N‐cadherin in primary colon cancer progression, which may be similar to the effect discovered by others in breast cancer cell lines, where coexpressed N‐cadherin can exert a dominant function over E‐cadherin's adhesive function and thus promote tumor invasiveness. © 2004 Wiley‐Liss, Inc.
Preoperative hCGβ and CA 72‐4 are prognostic factors in gastric cancer
Authors: Louhimo Johanna , Kokkola Arto , Alfthan Henrik , Stenman Ulf‐Håkan , Haglund Caj
Published: May 2004

In gastric cancer, the role of tumour markers in assessment of prognosis is unconfirmed. In our study, we evaluated the prognostic significance of serum tumour markers carcinoembryonic antigen (CEA), CA 19‐9, CA 72‐4, CA 242 and free β subunit of human chorionic gonadotropin (hCGβ) in gastric cancer. Preoperative serum samples were obtained from 146 patients with gastric cancer, including 29 with stage I, 11 with stage II, 42 with stage III and 64 patients with stage IV cancer. Quantitation of CEA, CA 19‐9, CA 72‐4 and CA 242 in serum was performed with commercial assays. HCGβ was measured with an in‐house immunofluorometric assay based on monoclonal antibodies specific for the free β‐subunit of hCG. Survival analysis was performed with Kaplan‐Meier life‐tables and log‐rank test, and with multivariate Cox regression analysis. Disease‐specific cumulative 2‐year survival rate was 40%. Serum levels of CEA, CA 72‐4, CA 242 and hCGβ showed significant correlation with stage (p<0.027); for CA 19‐9 the association was of borderline significance (p=0.056). Of the studied markers, CA 19‐9, CA 72‐4, CA 242 and hCGβ were found to be prognostic factors in univariate analysis (p< 0.022). In multivariate analysis, stage had the statistically most significant association with prognosis followed by hCGβ, tumour histology according to the Laurén classification and by CA 72‐4. In gastric cancer, tumour markers hCGβ and CA 72‐4 are independent prognostic factors in addition to stage and histological type of the tumour. © 2004 Wiley‐Liss, Inc.
Association between female breast cancer and cutaneous melanoma
Authors: Goggins William , Gao Wei , Tsao Hensin
Published: May 2004

Epidemiologic studies have provided suggestive evidence of a link between cutaneous melanoma (CM) and breast cancer (BC). Moreover, carriers of mutations in the breast cancer predisposition gene, BRCA2, have an increased risk of melanoma while carriers of mutations in the melanoma susceptibility gene, CDKN2A, exhibit a higher than expected risk of breast cancer. These findings raise the possibility that pathways involved in the development of CM and BC overlap and that survivors of one cancer may be prone to develop the other. To this end, we set out to determine if survivors of female BC in the Surveillance, Epidemiology and End Result (SEER) database are at increased risk for CM and vice versa. We followed female BC patients registered in the 1973—1999 SEER database for development of a second CM and female CM patients for the development of a second BC. The expected number of cases was then compared to the observed number of cases using standardized incidence ratios. Overall, we found a modest but statistically significant increased risk of CM among female BC survivors and vice versa. Among young BC patients, we observed a 46% elevated risk of a second CM. Women who underwent radiation therapy exhibited a 42% increased risk for CM. The risks of BC among female CM survivors and CM among BC survivors were also elevated, albeit to a much lesser degree (overall, 11% and 16%, respectively). We found a mutual association between female BC and CM. The elevated risk for CM, especially among younger BC patients, suggests that the genetic observations from high‐risk groups may also be operative at a much lower level in the general BC population. © 2004 Wiley‐Liss, Inc.
DNA binding sites for putative methylation boundaries in the unmethylated region of the BRCA1 promoter
Authors: Butcher Darci T. , Mancini‐DiNardo Debora N. , Archer Trevor K. , Rodenhiser David I.
Published: May 2004

Changes in DNA methylation patterns are frequently observed in human cancers and are associated with a decrease in tumor suppressor gene expression. Hypermethylation of the BRCA1 promoter has been reported in a portion of sporadic breast tumours that correspond to a reduction in BRCA1 transcription and expression. Questions remain concerning the maintenance of methylation free zones in promoter regions of tumor suppressor genes in normal tissues. Sodium bisulfite based analysis of the BRCA1 promoter defines a methylation free zone in normal breast tissue that starts 650 bp upstream of the transcription start site and extends for 1.4 kb through most of the BRCA1 CpG island. We provide data implicating 2 proteins, Sp1 and CTCF, in the maintenance of this methylation‐free zone. Both of these proteins have been shown to function as methylation boundaries in other genes. Four Sp1 sites have been identified in the BRCA1 promoter by gel shift assays. In vivo binding of Sp1 has been confirmed at 2 of these sites in the BRCA1 promoter and at 2 CTCF sites that flank the unmethylated region. Our data suggest that these DNA binding sites for Sp1 and CTCF may act as boundary elements that are important in maintaining genomic integrity by delineating the normal methylation free BRCA1 promoter. Inactivation or disruption of these boundaries may facilitate an epigenetic “hit”, in this case DNA methylation, leading to BRCA1 downregulation and contributing to tumorigenesis, particularly the genesis of sporadic breast tumours. © 2004 Wiley‐Liss, Inc.
Dietary calcium, vitamin D, VDR genotypes and colorectal cancer
Authors: Slattery Martha L. , Neuhausen Susan L. , Hoffman Michael , Caan Bette , Curtin Karen , Ma Khe Ni , Samowitz Wade
Published: May 2004

The vitamin D receptor (VDR) may importantly modulate risk of colorectal cancer either independently or in conjunction with calcium and vitamin D intake. We evaluate the association between calcium, vitamin D, dairy products, and VDR polymorphisms in 2 case‐control studies of colon and rectal cancer (n = 2,306 cases and 2,749 controls). Dietary intake was evaluated using a detailed diet history questionnaire. Two VDR polymorphisms were evaluated: an intron 8 Bsm 1 polymorphism and a 3′ untranslated region poly‐A length polymorphism (designated S for short and L for long). The SS genotype reduced risk of colorectal cancer for men (odds ratio [OR] = 0.71; 95% confidence interval [CI] = 0.55–0.92). High levels of calcium intake reduced risk of rectal cancer in women (OR = 0.39; 95% CI = 0.24–0.64) but were not associated with rectal cancer in men (OR = 1.02; 95% CI = 0.66–1.56). Similar reduced rectal cancer risk among women was observed at high levels of vitamin D (OR = 0.52; 95% CI = 0.32–0.85) and low‐fat dairy products (OR = 0.61; 95% CI = 0.39–0.94). High levels of sunshine exposure reduced risk of rectal cancer among those diagnosed when <60 years of age (OR = 0.62, 95% CI = 0.42–0.93). Examination of calcium in conjunction with VDR genotype showed that a significant 40% reduction in risk of rectal cancer was observed for the SS or BB VDR genotypes when calcium intake was low (p interaction = 0.01 for calcium interaction). For colon cancer, high levels of dietary intake of calcium, vitamin D, and low‐fat dairy products reduced risk of cancer for the SS or BB VDR genotypes, although the p for interaction was not statistically significant. These data support previous observations that high levels of calcium and vitamin D reduce risk of rectal cancer and provide support for a weak protective effect for the SS and BB VDR genotypes. The risk associated with VDR genotype seems to depend upon the level of dietary calcium and vitamin D and tumor site. © 2004 Wiley‐Liss, Inc.
Vaccination with tumor cell lysate‐pulsed dendritic cells augments the effect of IFN‐β gene therapy for malignant glioma in an experimental mouse intracranial glioma
Authors: Saito Ryuta , Mizuno Masaaki , Nakahara Norimoto , Tsuno Takaya , Kumabe Toshihiro , Yoshimoto Takashi , Yoshida Jun
Published: May 2004

Interferon‐β (IFN‐β) has been used as an antitumor drug against human glioma, melanoma and medulloblastoma since the 1980s. Recently, we developed a new gene therapy using the IFN‐β gene against malignant gliomas and then began clinical trials in 2000. Since stimulation of immune system was one mechanism of antitumor effect induced by IFN‐β gene therapy, we hypothesized that combination of IFN‐β gene therapy with immunotherapy might increase its effectiveness. In the present study, we tested whether combination therapy with IFN‐β gene therapy and immunotherapy using tumor cell lysate‐pulsed dendritic cells (DCs) would increase the efficacy of IFN‐β gene therapy. In an experimental mouse intracranial glioma (GL261), which cannot be cured by either IFN‐β gene therapy or DC immunotherapy alone, IFN‐β gene therapy following DC immunotherapy resulted in a significant prolongation in survival of the mice. Moreover, when this combination was performed twice, 50% of treated mice survived longer than 100 days. Considering these results, this combination therapy may be one promising candidate for glioma therapy in the near future. © 2004 Wiley‐Liss, Inc.
Extensive analysis of the 7q31 region in human prostate tumors supports TES as the best candidate tumor suppressor gene
Authors: Chêne Laurent , Giroud Christiane , Desgrandchamps François , Boccon‐Gibod Laurent , Cussenot Olivier , Berthon Philippe , Latil Alain
Published: May 2004

Loss of heterozygosity (LOH) on chromosome arm 7q31 is found in many prostate tumors. Such alterations are generally associated with inactivation of tumor suppressor genes. It has been shown previously that the main region of LOH at 7q31 spans the interval between the D7S486 and D7S2460 microsatellite loci, which contains several candidate tumor suppressor genes (TSG) such as TES, CAV2, CAV1, MET, CAPZA2, ST7 and WNT2. We tested 41 human sporadic prostate tumors for 7q31 LOH by using 5 polymorphic markers overlapping the critical region and used a real‐time reverse transcriptase‐polymerase chain reaction (RT‐PCR) assay to study the expression of the 7 candidate TSGs located in this genomic region. We found that CAV1, CAV2, MET and TES mRNA expression was lower in prostate tumors than in normal prostate tissues. Our immunohistochemical results and previously published data on the compartmental expression of these messenger RNAs in stromal and epithelial cells suggest that TES is the best candidate tumor suppressor gene at 7q31. © 2004 Wiley‐Liss, Inc.
Possible association between HPV16 E7 protein level and cytokeratin 19
Authors: Favia Gianfranco , Kanduc Darja , Lo Muzio Lorenzo , Lucchese Alberta , Serpico Rosario
Published: May 2004

The mechanisms employed by human papillomaviruses (HPVs) to control the replication of the viral genome and the expression of the viral genes are extremely complex and further complicated by the fact that the viral life cycle is intricately tied to the differentiation program of its host epithelial tissue. Indeed, HPV‐induced immortalization of keratinocytes and disruption of the normal cytokeratin (CK) expression pattern progress pari passu during the stepwise process that preludes to squamous cell carcinoma. In our study, we have analyzed the interaction occurring between HPV type 16 E7 mRNA and the intermediate cytokeratin filaments 7 and 19 and report data in favor of a possible association between HPV16 E7 protein level and CK19. © 2004 Wiley‐Liss, Inc.
Genetic epidemiology of cancer: From families to heritable genes
Authors: Hemminki Kari , Rawal Rajesh , Chen Bowang , Bermejo Justo Lorenzo
Published: May 2004

A reliable determination of familial risks for cancer is important for clinical counseling, prevention and understanding cancer etiology. Family‐based gene identification efforts may be targeted if the risks are well characterized and the mode of inheritance is identified. Medically verified data on familial risks have not been available for all types of cancer but they have become available through the use of the nationwide Swedish Family‐Cancer Database, which includes all Swedes born in 1932 and later with their parents, totaling over 10 million individuals. Over 150 publications have emanated from this source. The familial risks of cancer have been characterized for all main cancers and the contribution of environmental and heritable effects to the familial aggregation has been assessed. Furthermore, the mode of inheritance has been deduced by comparing risks from parental and sibling probands. Examples are shown on familial clustering of cancers, for which heritable susceptibility genes are yet unknown, such as squamous cell carcinoma of the skin, intestinal carcinoids, thyroid papillary tumors, brain astrocytomas and pituitary adenomas. Some common cancers, such as lung and kidney cancers, appear to show an early‐onset recessive component because familial risks among siblings are much higher than those in families where parents are probands. Many of the cancer sites showing high familial risks lack guidelines for clinical counseling or action level. In conclusion, we recommend that any future gene identification efforts, either using linkage or association designs, devise their strategies based on data from family studies. Clinical genetic counseling would benefit from reviewing established familial risks on all main types of cancer. © 2004 Wiley‐Liss, Inc.
Downregulation of KiSS‐1 expression is responsible for tumor invasion and worse prognosis in gastric carcinoma
Authors: Dhar Dipok Kumar , Naora Hiroyuki , Kubota Hirofumi , Maruyama Riruke , Yoshimura Hiroshi , Tonomoto Yasuhito , Tachibana Mitsuo , Ono Takashi , Otani Hiroki , Nagasue Naofumi
Published: May 2004

KiSS‐1 is a promising candidate tumor‐suppressor gene and may play a key role in the metastatic cascade. The expression profile and the role of KiSS‐1 in cancer progression are largely unknown in most of the cancers, including gastric cancer. In this study, KiSS‐1 expression was evaluated by RNase protection assay and localization was done by in situ hybridization in 40 gastric cancers and their adjacent normal gastric mucosa. For comparison with clinicopathologic characteristics and patient prognosis, all patients were divided into 2 groups having high and low KiSS‐1 expression by using the median as the cutoff value of KiSS‐1 expression as determined by the RNase protection assay. Gastric cancers with low KiSS‐1 had frequent venous invasion, distant metastasis and tumor recurrence. Accordingly, patients with low KiSS‐1‐expressing tumors had a significantly worse overall and disease‐free survival. In multivariate analysis, KiSS‐1 became the strongest independent prognostic factor among the conventional prognosticators for gastric cancer patients. Collectively, these findings suggest that KiSS‐1 may play a crucial role in gastric cancer invasion and could be a useful target for therapeutic intervention. © 2004 Wiley‐Liss, Inc.
ZNF143 activates gene expression in response to DNA damage and binds to cisplatin‐modified DNA
Authors: Ishiguchi Hiroshi , Izumi Hiroto , Torigoe Takayuki , Yoshida Yoichiro , Kubota Hiroshi , Tsuji Sadatoshi , Kohno Kimitoshi
Published: May 2004

We have identified a cisplatin‐inducible gene, the mitochondrial ribosomal protein S11 (MRP S11) gene, by means of mRNA differential display. Functional analysis of the MRP S11 promoter showed that a Staf binding site in the promoter is required for both basal promoter activity and cisplatin‐inducible activity. We also found that Staf binding activity is significantly increased in nuclear extracts from cells treated with cisplatin. ZNF 143 and ZNF 76 are human homologues of the Xenopus transcriptional activator, Staf. ZNF 143 expression is induced by cisplatin but ZNF 76 expression is not. However, ZNF 143 expression is not induced by transplatin, which is clinically ineffective. ZNF143 is an inducible gene by other DNA damaging agents such as γ‐irradiation, etoposide and adriamycin. ZNF 143 also binds preferentially to cisplatin‐modified DNA. These results suggest that ZNF 143 participates in cellular responses to DNA damage. © 2004 Wiley‐Liss, Inc.
Pegylated polystyrene particles as a model system for artificial cells
Authors: Meng Fenghua , Engbers Gerard H. M. , Gessner Andrea , Müller Reiner H. , Feijen Jan
Published: May 2004

Pegylated polystyrene particles (PS‐PEG) were prepared as a model system for artificial cells, by modification of carboxyl polystyrene particles (PS‐COOH) with homo‐ and hetero‐bifunctional polyethylene glycols (PEG, MW 1500, 3400, and 5000) containing an amino end group for immobilization and an amino, hydroxyl, or methoxy end group that is exposed at the surface after immobilization. Protein adsorption from human plasma dilutions (85 v %) onto PS‐PEG with a PEG surface concentration higher than 40 pmol/cm2 was reduced up to 90–95% compared with protein adsorption onto PS‐COOH with a final protein surface concentration of ∼30 ng/cm2. Two‐dimensional gel electrophoresis analyses showed that 30% of the total amount of adsorbed proteins onto PS‐PEG are dysopsonins (i.e., nonadhesive proteins like albumin and apolipoproteins). For PS‐COOH, <15% of the amount of adsorbed proteins are dysopsonins. In addition, the generation of terminal complement compound (TCC) by PS‐PEG particles with a PEG surface concentration lower than ∼55 pmol/cm2 is not significant. The low protein adsorption, the relatively high percentage of adsorbed dysopsonins, and the low level of complement activation may prevent the uptake of PS‐PEG by the mononuclear phagocytic system (MPS) in vivo. Moreover, PS‐PEG (PEG surface concentration > ∼35 pmol/cm2) shows minimal interaction with cultured human umbilical vein endothelial cells (HUVEC), which mimics the endothelial lining of the blood vessel wall. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 70A: 97–106, 2004
PLA and PLGA microspheres of β‐galactosidase: Effect of formulation factors on protein antigenicity and immunogenicity
Authors: Stivaktakis N. , Nikou K. , Panagi Z. , Beletsi A. , Leondiadis L. , Avgoustakis K.
Published: May 2004

The entrapment of β‐galactosidase (Escherichia coli) in PLA and PLGA microspheres using a double emulsion technique resulted to significant reduction of protein antigenicity. The extent of antigenicity loss depended on the conditions of microsphere preparation. Most of antigenicity loss occurred on the first emulsification step. Only the effects of microsphere preparation factors having an important influence on protein antigenicity, such as the type of organic phase (polymer solvent) and homogenization, could be predicted (on a qualitative basis) by antigenicity data obtained after the first emulsification step. The type of polymer and polymer solvent used to prepare the microspheres affected β‐galactosidase immunogenicity. The PLA microspheres prepared using ethyl acetate was the most immunogenic microsphere formulation, eliciting similar total antibody responses as the alum formulation of β‐gal. This formulation was the only microsphere formulation that induced an IgG1/IgG2a ratio lower than 1, indicating an immune response biased towards a Th1 type. The results obtained indicate that large protein molecules with complex tertiary structure such as β‐galactosidase can be entrapped in PLA and PLGA microspheres with retention of protein immunogenic potential, providing that appropriate conditions of microsphere preparation are applied, and that the formulation of microspheres might influence the Th1/Th2 type of immune response against the encapsulated antigen. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 70A: 139–148, 2004
Influence of orthopedic particulate biomaterials on inflammation and synovial microcirculation in the murine knee joint
Authors: Zysk Stefan P. , Gebhard Harry , Plitz Wolfgang , Buchhorn G.H. , Sprecher Christoph M. , Jansson Volkmar , Messmer Konrad , Veihelmann Andreas
Published: May 2004

The purpose of the present study was to examine changes in the synovial microcirculation as well as synovial tissue responses to exposure to titanium, polymethylmethacrylate (PMMA), ceramic (Al2O3), cobalt–chromium alloy (Co–Cr), and polyethylene (PE) particles in an in vivo model. The particulate biomaterials were injected into the left knee joint of female Balb/c mice and assessment of the synovial microcirculation using intravital fluorescence microscopy as well as histological evaluation of the synovial tissue response were performed on day 7 after particle administration. Intravital microscopic measurements revealed that all tested biomaterials caused significantly (p < 0.05) enhanced leukocyte–endothelial cell interactions and an increase of functional capillary density compared to controls. In the histological examination PMMA, Al2O3, PE, and Co–Cr particles provoked significantly (p < 0.05) enhanced inflammatory tissue responses in comparison to tissue from control animals. Titanium particles showed significantly (p < 0.05) less leukocyte–endothelial cell interactions than the other particulate biomaterials and caused significantly (p < 0.05) minor membrane thickening compared to PE and PMMA particles. In conclusion, all tested particulate biomaterials were capable of inducing inflammatory responses in the present study. Our data suggest that titanium particles may cause less leukocyte activation and inflammatory tissue responses than other particulate biomaterials used in total joint arthroplasty. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 71B: 108–115, 2004
Differential activation of ERK1,2 MAP kinase signaling pathway in mesenchymal stem cell from control and osteoporotic postmenopausal women
Authors: Rodríguez J. Pablo , Ríos Susana , Fernández Mireya , Santibañez J. Francisco
Published: May 2004

Osteoblasts, the cells responsible for bone formation, derive from mesenchymal stem cells (MSCs) in bone marrow. To acquire a new cell phenotype, uncommitted MSCs must undergo several proliferation and differentiation changes. Although, it is known that extracellular signal‐regulated protein kinases (ERKs) mitogen‐activated protein (MAP) kinase pathway signaling is involved in the proliferation and differentiation processes, the role of ERKs in osteogenic differentiation it is controversial, at present. In addition, the function that ERK could play in MSCs derived from osteoporotic patients it is not well documented. In this study, we analyze whether previously observed differences in the dynamic response of MSCs from normal and osteoporotic postmenopausal women can be explained by changes in the activation of this signal transduction pathway. Levels of ERK phosphorylation and their correlation with osteogenic differentiation were evaluated in cultures of MSCs derived from osteoporotic postmenopausal women and “healthy” controls. The results show that, under basal conditions, MSCs derived from osteoporotic donors show a level of ERK phosphorylation 2.5 times higher than MSCs derived from control donors. The addition of the osteogenic stimulus only slightly increases the p‐ERK level in cells derived from osteoporotic donors, and is higher in cells derived from control women. Important differences in the ability of PD98059 to inhibit phosphorylation of ERK in both types of cells were also observed, as well as the effect that this inhibition produced on calcium deposition. We conclude that the MAP kinase pathway signaling is differentially activated in MSCs derived from osteoporotic postmenopausal women. The high p‐ERK levels in MSC derived from osteoporotic donors could determine the unresponsiveness of these cells to the osteogenic differentiation stimulus. © 2004 Wiley‐Liss, Inc.
Consultation and interprofessional collaboration: Modeling for the future
Authors: Arredondo Patricia , Shealy Craig , Neale Michael , Winfrey LaPearl Logan
Published: May 2004

Consultation and interprofessional collaboration by psychologists occur with individuals, groups, programs, and organizations. The practice of consultation and interprofessional collaboration involves interdisciplinary relationships, preparation, and advanced skill development within specialty areas of psychology (e.g., clinical, counseling, industrial‐organizational, and school). The Workgroup on Consultation and Interdisciplinary Relationships engaged in a planning process at the Competencies Conference: Future Directions in Education and Credentialing in Professional Psychology to address fundamental issues regarding consultation and interprofessional collaboration in professional psychology. The Workgroup articulated working definitions, consensus points about psychologists as consultants and interprofessional collaborators, a consulting and interprofessional competency blueprint for preparation and assessment strategies, and future directions. This is one of a series of articles published in this issue of the Journal of Clinical Psychology. Several other articles that resulted from the Competencies Conference will appear in Professional Psychology: Research and Practice and The Counseling Psychologist. © 2004 Wiley Periodicals, Inc. J Clin Psychol.
Community resilience and the impact of stress: Adult response to Israel's withdrawal from Lebanon
Authors: Kimhi Shaul , Shamai Michal
Published: May 2004

Against the background of the Israeli withdrawal from Lebanon, we investigated the relationship between perceived community resilience and the effect of stress and life satisfaction. The research sample included 741 adults, aged 18–85. The participants were divided into four groups, three of which live close to the Israel–Lebanon border and were directly exposed to the threat created by war and terror. The fourth group was considered as a control group and included subjects from the central region of Israel, who were not directly exposed to the war with Lebanon and to the possible outcomes of withdrawal. Questionnaires were distributed to the participants immediately after the withdrawal from Lebanon and were completed by them between 1 and 3 weeks after the withdrawal. The items were designed to measure perceived community resilience, the effects of stress, and life satisfaction, and demographic background. The results show that the level of threat has a significant impact on community resilience, namely, that living in situations with a high level of threat over a long period of time results in a lower level of community resilience. In addition, community resilience serves as a partial mediator between the level of threat and the effect of stress and life satisfaction. The results highlight the importance of perceived community resilience as an individual resource for coping with the threat created by war and terror, thereby connecting between micro‐ and macro‐levels in events related to political violence. © 2004 Wiley Periodicals, Inc. J Comm Psychol 32: 439–451, 2004.
PDGF and FGF induce focal adhesion kinase (FAK) phosphorylation at Ser‐910: Dissociation from Tyr‐397 phosphorylation and requirement for ERK activation
Authors: Hunger‐Glaser Isabel , Fan Robert S. , Perez‐Salazar Eduardo , Rozengurt Enrique
Published: May 2004

A rapid increase in the tyrosine phosphorylation of focal adhesion kinase (FAK) has been extensively documented in cells stimulated by multiple signaling molecules, but very little is known about the regulation of FAK phosphorylation at serine residues. Stimulation of Swiss 3T3 cells with platelet‐derived growth factor (PDGF) promoted a striking increase in the phosphorylation of FAK at Ser‐910, as revealed by site‐specific antibodies that recognized the phosphorylated state of this residue. FAK phosphorylation at Ser‐910 could be distinguished from that at Tyr‐397 in terms of dose‐response relationships and kinetics. Furthermore, the selective phosphoinositide 3‐kinase (PI 3‐kinase) inhibitors wortmannin and LY 294002 abrogated FAK phosphorylation at Tyr‐397 but did not interfere with PDGF‐induced FAK phosphorylation at Ser‐910. Conversely, treatment with U0126, a potent inhibitor of MEK‐mediated ERK activation, prevented FAK phosphorylation at Ser‐910 induced by PDGF but did not interfere with PDGF‐induced FAK phosphorylation at Tyr‐397. These results were extended using growth factors that either stimulate, fibroblast growth factor (FGF), or do not stimulate (insulin) the ERK pathway activation in Swiss 3T3 cells. FGF but not insulin promoted a striking ERK‐dependent phosphorylation of FAK at Ser‐910. Our results indicate that FAK phosphorylation at Tyr‐397 and FAK phosphorylation at Ser‐910 are induced in response to PDGF stimulation through different signaling pathways, namely PI 3‐kinase and ERK, respectively. © 2004 Wiley‐Liss, Inc.
Chimeric molecule IL‐6/soluble IL‐6 receptor is a potent mitogen for fetal hepatocytes
Authors: Zvibel Isabel , Brill Shlomo , Kariv Revital , Traister Alexandra , Golan Talia , Chebath Judith , Halpern Zamir , Revel Michel , Oren Ran
Published: May 2004

A novel recombinant molecule, termed IL‐6c and consisting of a chimera of interleukin 6 (IL‐6) and its soluble receptor is extremely potent in stimulating proliferation of hematopoietic progenitors. We investigated the effect of the IL‐6c on the proliferation and differentiation of E14 fetal hepatocytes. IL‐6c, in a dose‐dependent manner, stimulated proliferation of E14 fetal rat hepatocytes. Adult hepatocyte mitogens together with IL‐6c showed no further effect on proliferation. Hematopoietic stem cells mitogens SCF and flt3 ligand (FL) were also mitogenic for fetal hepatocytes, but did not further enhance the effect of IL‐6c on cell proliferation. IL‐6c decreased expression of fetal markers α‐fetoprotein (AFP) and gamma‐glutamyltranspeptidase, and induced expression of adult enzyme glucose‐6‐phosphatase (Gluc‐6‐P) in E14 hepatocytes. On the other hand, IL‐6c strongly reduced, in a dose‐dependant manner, expression of albumin and tyrosine aminotransferase (TAT). However, when the cells were grown for 3 days with IL‐6c, and IL‐6c was removed for the next 5 days, expression of albumin and TAT returned to levels found in control cultures. In conclusion, IL‐6c stimulated proliferation and affected gene expression in fetal hepatocytes in culture. © 2004 Wiley‐Liss, Inc.
Role of LRP in TGFβ2‐mediated neuronal uptake of Aβ and effects on memory
Authors: Harris‐White Marni E. , Balverde Zerlinde , Lim Giselle P. , Kim Peter , Miller Sheryl A. , Hammer Heidi , Galasko Doug , Frautschy Sally A.
Published: May 2004

There is increasing evidence that soluble amyloid‐β peptide (Aβ) uptake into neurons is an early event in the pathogenesis of Alzheimer's disease (AD). Identification of the early events leading to neuronal dysfunction is key to developing therapeutic strategies, but relative roles of receptors and factors modulating uptake are poorly understood. Studies have shown that transforming growth factor β (TGFβ), particularly TGFβ2, can influence the targeting of Aβ to cells in vitro. TGFβ2 can target Aβ to neurons in organotypic hippocampal slice cultures (OHSC). We examine a specific mechanism for TGFβ2‐mediated targeting of Aβ to neurons. The receptor‐associated protein (RAP), a low‐density lipoprotein receptor‐related protein (LRP) antagonist, can attenuate the cellular targeting of Aβ both in vitro and in vivo and prevent Aβ/TGFβ2‐induced memory retention deficits. Using both in vitro and in vivo methods, we identify LRP as playing a role in TGFβ2‐mediated Aβ uptake, neurodegeneration, and spatial memory impairment. © 2004 Wiley‐Liss, Inc.
Activation of adenosine A2A receptor protects sympathetic neurons against nerve growth factor withdrawal
Authors: Ramirez Servio H. , Fan Shongshan , Maguire Casey A. , Perry Seth , Hardiek Kathy , Ramkumar Vickram , Gelbard Harris A. , Dewhurst Stephen , Maggirwar Sanjay B.
Published: May 2004

Adenosine mediates a range of effects in the central nervous system (CNS), including the promotion of neuronal survival, but its actions on sympathetic neurons are less well characterized. We therefore sought to understand the role of endogenous adenosine in contributing to the survival of neurotrophin‐dependent sympathetic neurons. Rat superior cervical ganglion (SCG) cultures were maintained in the continuous presence of nerve growth factor (NGF) and then exposed to adenosine deaminase (ADA), to deplete endogenous adenosine. This resulted in a marked increase in cellular apoptosis, to a level that approximated the effect of NGF withdrawal. Furthermore, the addition of exogenous adenosine to NGF‐deprived SCG neurons resulted in enhanced cell survival. Analysis of adenosine receptor (AR) subtypes on these neurons, using real‐time RT‐PCR and receptor binding analyses, revealed that the A2A receptor was the major subtype present. Accordingly, the A2A receptor agonist CGS21680 significantly reduced both ADA‐induced and NGF‐withdrawal‐induced neuronal apoptosis, whereas the A1 receptor agonist R‐PIA had no such effect. The survival‐promoting effect of CGS21680 was eliminated when cells were coincubated with a molar excess of an A2A receptor antagonist. Finally, follow‐up experiments revealed that CGS21680 prevented the induction of early apoptotic events, such as changes in mitochondrial integrity and caspase activation, and that it also triggered an increase in ERK activation, which was essential for neurotrophin‐independent cell survival. Taken together, these findings provide evidence that endogenous adenosine may be important in mediating protection of sympathetic neurons and that it may act via the A2A receptor subtype. © 2004 Wiley‐Liss, Inc.
Effect of the peroxisome proliferator‐activated receptor β activator GW0742 in rat cultured cerebellar granule neurons
Authors: Smith Steven A. , Monteith Gregory R. , Robinson Jodie A. , Venkata Nagaraj Gopisetty , May Fiona J. , Roberts‐Thomson Sarah J.
Published: May 2004

The ligand‐activated transcription factor peroxisome proliferator‐activated receptor β (PPARβ) is present in the brain and is implicated in the regulation of genes with potential roles in neurotoxicity. We sought to examine the role of PPARβ in neuronal cell death by using the PPARβ ligand GW0742. Primary cultures of rat cerebellar granule neurons were prepared from 7‐day‐old pups. Reverse transcriptase‐polymerase chain reaction and in situ hybridization were used to verify that PPARβ mRNA was present in neurons. After 10–12 days in culture, the neuronal cells were incubated in the presence of GW0742, and cell death was measured with a lactate dehydrogenase release (LDH) assay. After 24 hr of exposure, PPARβ activation by GW0742 was not inherently toxic to cerebellar granule neurons. However, toxicity was observed after 48 hr, with cell death mediated via an apoptotic mechanism. In an effect opposite to that observed with PPARα‐activating ligands, PPARβ activation exhibited neuroprotective properties. Treatment with GW0742 significantly reduced cell death during a 12‐hr exposure to low‐KCl media. These results clearly reinforce very specific roles for the PPAR isoforms in neurons and suggest that PPARβ is worthy of further investigation regarding its potential role as a therapeutic target in neurodegenerative states. © 2004 Wiley‐Liss, Inc.
Cyclic AMP/protein kinase a signal attenuates Ca2+‐induced fibroblast growth factor‐1 synthesis in rat cortical neurons
Authors: Kinukawa Hideki , Jikou Takahiro , Nitta Atsumi , Furukawa Yoshiko , Hashimoto Manabu , Fukumitsu Hidefumi , Nomoto Hiroshi , Furukawa Shoei
Published: May 2004

Fibroblast growth factor (FGF)‐1 is increased in particular brain regions after birth, suggesting an involvement of some regulatory neuronal circuits. To address the neuronal activity responsible for FGF‐1 synthesis, effects of various neurotransmitter receptor activation on cellular FGF‐1 content were examined using cultured rat cortical neurons. Histamine, glutamate, carbachol, serotonin or γ‐aminobutyric acid (GABA) caused an increase of FGF‐1 content. Because this effect was mimicked by (1) N‐methyl‐D‐aspartate, a glutamatergic agonist; (2) Ca2+ ionophore; (3) depolarization with high concentration of KCl, but was abolished in Ca2+‐free medium, Ca2+ influx was thought to trigger FGF‐1 synthesis. Such Ca2+‐mediated enhancement of FGF‐1 synthesis, however, did not occur in the presence of norepinephrine (NE), but was restored by KT‐5720, an inhibitor of protein kinase A (PKA), suggesting an interplay between Ca2+‐activated and cAMP/PKA signals for neuronal FGF‐1 synthesis. This mechanism was proved to function in vivo by stimulation of FGF‐1 expression in neurons of the cerebral cortex after intracerebral administration of propranolol, an antagonist of adrenergic β receptors. This demonstrates that FGF‐1 synthesis is essentially upregulated by Ca2+ influx through excitatory neuronal activities, but such an effect is abolished by neurotransmission that evokes cAMP/PKA signals. FGF‐1 produced is thought to act on establishment and maintenance of particular neuronal circuits in the brain, which may be one of the ways neurotransmitters regulate brain function. © 2004 Wiley‐Liss, Inc.
Activity‐regulated cytoskeleton‐associated protein arc is targeted to dendrites and coexpressed with mu‐opioid receptors in postnatal rat caudate‐putamen nucleus
Authors: Wang Hong , Pickel Virginia M.
Published: May 2004

Dendritic expression of the activity‐regulated cytoskeleton‐associated protein (Arc) is dramatically enhanced by increased synaptic activity in adult brain. We used immunocytochemical electron microscopy to determine whether the subcellular localization of Arc in developing dendrites corresponds to the peak period of synaptogenesis in the postnatal rat caudate‐putamen nucleus (CPN). The distribution was compared with that of mu‐opioid receptors (MORs), whose localization in dendritic spines closely parallels excitatory synapse formation during postnatal development (Wang et al. [2003] Neuroscience 118:695–708). Sections were processed for immunocytochemical detection of antisera against Arc or MORs at the beginning (postnatal day 15; P15) and the end (P30) of the peak period of synaptogenesis in rat CPN. At P15, immunolabeling for Arc showed a punctate distribution in the cytoplasm of dendritic shafts, some of which was associated with polyribosomes. In some spiny dendrites, Arc immunoreactivity was more intensely localized in putative spines than in their parental dendrites, whereas, in other spiny dendrites, Arc labeling was restricted in the shafts. Many dendritic shafts and spines also showed immunoreactivity for MORs, although dually labeled spines were less numerous than the shafts. At P30, the proportion of singly and dually labeled spines significantly increased from 2.0% to 7.5% and from 9.5% to 21%, respectively. Arc labeling in spines was more detectable beneath the postsynaptic density or at extrasynaptic sites on the plasma membrane. Our results suggest a correlation between Arc expression in dendritic spines during postnatal development and the onset of synaptogenesis in opioid‐responsive neurons in the rat CPN. © 2004 Wiley‐Liss, Inc.
Interplay among platelet‐activating factor, oxidative stress, and group I metabotropic glutamate receptors modulates neuronal survival
Authors: Zhu Peimin , DeCoster Mark A. , Bazan Nicolas G.
Published: May 2004

Platelet‐activating factor (PAF) is a potent phospholipid messenger in the nervous system that participates in synaptic plasticity and in pathologic processes, including neurodegeneration. Oxidative stress plays important roles in neuronal cell death. To define the interaction between PAF and oxidative radicals in neuronal death, we studied the effects of PAF in the presence of oxidative radicals in primary neurons in culture. Exogenous PAF (50 μM) caused PAF receptor‐independent injury to neurons. A nonneurotoxic PAF concentration (500 nM) potentiated neuronal death caused by hydrogen peroxide as determined by lactate dehydrogenase (LDH) assay, Hoechst staining, and TUNEL analysis, but it did not potentiate neuronal death caused by menadione, a superoxide donor, or by the nitric oxide donors 3‐morpholino‐sydnonimine (SIN‐1) and sodium nitroprusside (SNP). This potentiation of the hydrogen peroxide effect was selectively blocked by a PAF membrane‐receptor antagonist, BN52021 (5 μM). The neurotoxic effect of PAF and hydrogen peroxide was also completely blocked by ebselen and partially decreased by pretreatment with (S)‐3,5‐dihydroxyphenylglycine (DHPG), a group I metabotropic glutamate receptor (mGluR) agonist. This study suggests that PAF‐receptor antagonists may be useful for neuroprotection. A similar effect might also be obtained with group I mGluR agonists, probably by way of a different underlying mechanism. © 2004 Wiley‐Liss, Inc.