bluuebunny/crossref_metadata_2025 huggingface dataset:
Control of continuous polyhydroxybutyrate synthesis using calorimetry and flow cytometry
Authors: Maskow Thomas , Müller Susann , Lösche Andreas , Harms Hauke , Kemp Richard
Published: October 2005

The substrate‐carbon flow can be controlled in continuous bioreactor cultures by the medium composition, for example, by the C/N ratio. The carbon distribution is optimal when a maximum fraction flows into the desired product and the residual is just sufficient to compensate for the dilution of the microbial catalyst. Undershooting of the latter condition is reflected immediately by changes in the Gibbs energy dissipation and cellular states. Two calorimetric measurement principles were applied to optimize the continuous synthesis of polyhydroxybutyrate (PHB) by Variovorax paradoxus DSM4065 during growth with constantly increasing supply rates of fructose or toxic phenol. Firstly, the changed slope of the heat production rate in a complete heat balanced bioreactor (CHB) indicated optimum carbon channeling into PHB. The extent of the alteration depended directly on the toxic properties of the substrate. Secondly, a flow through calorimeter was connected with the bioreactor as a “measurement loop.” The optimum substrate carbon distribution was indicated by a sudden change in the heat production rate independent of substrate toxicity. The sudden change was explained mathematically and exploited for the long‐term control of phenol conversion into PHB. LASER flow cytometry measurements distinguished between subpopulations with completely different PHB‐content. Populations grown on fructose preserved a constant ratio of two subpopulations with double and quadruple sets of DNA. Cells grown on phenol comprised a third subpopulation with a single DNA set. Rising phenol concentrations caused this subpopulation to increase. It may thus be considered as an indicator of chemostress. © 2005 Wiley Periodicals, inc.
Thermostabilization of porcine kidney D‐amino acid oxidase by a single amino acid substitution
Authors: Bakke Mikio , Setoyama Chiaki , Miura Retsu , Kajiyama Naoki
Published: October 2005

D‐amino acid oxidase (DAO) is of considerable practical importance, such as bioconversion and enzymatic assay. In this study, we succeeded in obtaining a thermostable mutant DAO from porcine kidney by a single amino acid substitution. This mutant enzyme, F42C, was stable at 55°C, while the wild‐type enzyme was stable only up to 45°C. The Km values of F42C for D‐amino acids was about half of those of the wild‐type enzyme. This mutant DAO with improved stability and affinity for its substrates is advantageous for the determination of D‐amino acids. © 2005 Wiley Periodicals, inc.
In silico analysis for transcription factors with Zn(II)2C6 binuclear cluster DNA‐binding domains in Candida albicans
Authors: Maicas Sergi , Moreno Inmaculada , Nieto Almudena , Gómez Micaela , Sentandreu Rafael , Valentín Eulogio
Published: October 2005

A total of 6047 open reading frames in the Candida albicans genome were screened for Zn(II)2C6‐type zinc cluster proteins (or binuclear cluster proteins) involved in DNA recognition. These fungal proteins are transcription regulators of genes involved in a wide range of cellular processes, including metabolism of different compounds such as sugars or amino acids, as well as multi‐drug resistance, control of meiosis, cell wall architecture, etc. The selection criteria used in the sequence analysis were the presence of the CysX2CysX6CysX5–16CysX2CysX6–8Cys motif and a putative nuclear localization signal. Using this approach, 70 putative Zn(II)2C6 transcription factors have been found in the genome of C. albicans. Copyright © 2005 John Wiley & Sons, Ltd.
Characterization of pathologic complete response after preoperative chemoradiotherapy in carcinoma of the esophagus and outcome after pathologic complete response
Authors: Rohatgi Pooja , Swisher Stephen G. , Correa Arlene M. , Wu Tsung‐T. , Liao Zhongxing , Komaki Ritsuko , Walsh Garrett L. , Vaporciyan Ara A. , Rice David C. , Roth Jack A. , Ajani Jaffer A.
Published: October 2005

BACKGROUND

The purpose of the current study was to test the hypothesis that a lower clinical TNM stage is associated with a higher rate of pathologic complete response (pathCR) in patients with esophageal carcinoma receiving preoperative chemoradiotherapy and to determine whether outcome after pathCR is related to clinical stage or treatment.

METHODS

Clinical parameters and surgical specimens of patients with esophageal carcinoma undergoing preoperative chemoradiotherapy were analyzed to identify predictors of pathCR. In patients with pathCR, predictors of overall survival (OS), disease‐free survival (DFS), and distant recurrence were studied.

RESULTS

Sixty‐nine (29%) of 235 patients achieved pathCR. In patients with American Joint Committee on Cancer (AJCC) Stage II carcinoma, the proportion achieving pathCR was significantly larger than that achieving <pathCR (65% vs. 35%; P = 0.03). The proportion of patients who received induction chemotherapy was higher in the pathCR group than in the <pathCR group (54% vs. 46%; P = 0.05). However, neither TNM classification, primary tumor location, histologic type, gender, therapy sequence, or radiation dose (45 grays [Gy] vs. 50.4 Gy) were found to have any influence on OS or DFS. The median OS from pathCR was significantly longer than that from <pathCR (133 mos vs. 34 mos; P = 0.002). Similarly, DFS was longer in the pathCR group than in the <pathCR (P = 0.001).

CONCLUSIONS

Patients with clinical AJCC Stage II esophageal carcinoma are more likely to achieve a pathCR after preoperative chemoradiotherapy than are those with Stage III carcinoma. Chemoradiotherapy as primary therapy for patients with Stage I esophageal carcinoma warrants investigation as a means to preserve their esophagus. Cancer 2005. © 2005 American Cancer Society.
Tobacco smoking, GSTP1 polymorphism, and bladder carcinoma
Authors: Cao Wei , Cai Lin , Rao Jian‐Yu , Pantuck Allan , Lu Ming‐Lan , Dalbagni Guido , Reuter Victor , Scher Howard , Cordon‐Cardo Carlos , Figlin Robert A. , Belldegrun Arie , Zhang Zuo‐Feng
Published: October 2005

BACKGROUND

Although cigarette smoking is considered a major risk factor for bladder carcinoma, little is known about the interaction between metabolic genes such as glutathione‐S‐transferase P1 and tobacco smoking in this process. GSTP1 may play a role in detoxification of tobacco‐related carcinogens.

METHODS

In this case–control study of 145 cases with bladder carcinoma (male:female = 7.5:1) and 170 noncancer controls (male:female = 3.7:1), the relation between genetic polymorphisms of GSTP1 and susceptibility to bladder carcinoma was investigated and the gene–environment interaction between tobacco smoking and GSTP1 polymorphism was evaluated. Epidemiological data were collected for all cases and controls by a standard questionnaire. Polymorphisms of GSTP1 were measured by polymerase chain reaction–restriction fragment length polymorphism. The logistic regression model in SAS was used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs).

RESULTS

Cigarette smoking was confirmed as a risk factor of bladder carcinoma with an OR of 3.1 (95% CI: 1.7–5.9) after controlling for potential confounding factors. The OR for pack‐years of smoking as a continuous variable was 2.4 (95% CI: 2.0–2.8). The ORs were 7.6 (95% CI: 1.18–49.51) for isoleucine/valine (Ile/Val) and 6.5 (95% CI: 1.01–41.56) for Ile/Ile when the homozygous Val/Val was considered as comparison group after adjusting for age, gender, race, and education. The adjusted OR for interaction between smoking and the GSTP1 (any Ile genotype) was 11.42 (95% CI: 0.53–248.15).

CONCLUSIONS

The results indicate that the Ile 105 allele is associated with an increased risk of bladder carcinoma and suggest that individuals who smoke and possess the Ile allele might be at increased risk for bladder carcinoma. Cancer 2005. © 2005 American Cancer Society.
Long remissions in hairy cell leukemia with purine analogs
Authors: Else Monica , Ruchlemer Rosa , Osuji Nnenna , Del Giudice Ilaria , Matutes Estella , Woodman Anthony , Wotherspoon Andrew , Swansbury John , Dearden Claire , Catovsky Daniel
Published: October 2005

BACKGROUND

Both pentostatin and cladribine have efficacy in hairy cell leukemia (HCL), but it is not known which agent achieves better results.

METHODS

We reviewed a series of 219 patients with HCL, with median follow‐up from diagnosis of 12.5 years (range 1.0 –34.6 yrs), treated with either pentostatin (n= 185) or cladribine (n= 34), to compare these agents and assess the potential for cure.

RESULTS

Overall response to pentostatin was 96% with a complete response (CR) in 81% and a median disease‐free survival (DFS) of 15 years. Response to first‐line cladribine was 100% with a CR in 82% and DFS of 11+ years. The relapse rates at 5 years and 10 years were 24% and 42%, respectively, with pentostatin, and 33% and 48% with cladribine. Survival at 10 years was respectively 96% and 100%. CR rates decreased with each sequential relapse through 69% to 45% (P≤ 0.001). Patients achieving CR after first‐line treatment had a significantly longer DFS (P= 0.00007) than those achieving a partial response; a similar result was seen after second‐line therapy (P= 0.00001). DFS also declined with sequential treatment (P= 0.00005).

CONCLUSION

We have shown equivalent efficacies for both agents in the treatment of HCL, with DFS showing no plateau. True cure in HCL remains elusive, but the addition of monoclonal antibodies may be beneficial. Our results suggest that achieving CR should remain the main goal of treatment. Cancer 2005. © 2005 American Cancer Society.
Effect of a pT0 cystectomy specimen without neoadjuvant therapy on survival
Authors: Volkmer Bjoern G. , Kuefer Rainer , Bartsch Georg , Straub Michael , de Petriconi Robert , Gschwend Juergen E. , Hautmann Richard E.
Published: October 2005

BACKGROUND

In some cases of radical cystectomy for bladder cancer, no residual tumor is found in the cystectomy specimen (the pT0 classification). The aim of this study was to evaluate the outcome of such patients in a large cystectomy series.

METHODS

All 900 patients with radical cystectomy and pelvic lymphadenectomy for TCC of the bladder in the period January 1986 to September 2003 who received no neoadjuvant therapy were included. Cystectomy specimens from 181 (20.1%) patients were graded as pT0. Complete follow‐up was obtained in all cases. Tumor‐specific survival (pT0 vs. pT+) was calculated with the Kaplan–Meier method and compared with the log‐rank test.

RESULTS

The rate of lymph node metastases in the pT0 group was 6.6%. pT0 status was found with Ta/is/1 in 36.8%, T2a in 41.8%, and T2b in 10.9%. The 169 patients with pT0pN0 tumors had 10‐year tumor‐specific survival rates of 91.0. There was no statistically significant survival benefit for pT0pN0 tumors compared with pT+pN0 tumors for maximal tumor classifications of pTa/pTis/pT1 and pT2b, but patients with a pT0T2apN0 tumor had a statistically significantly better tumor‐specific survival than those with a pT2apN0 tumor (P = 0.002). No patient with a pT0pN0 tumor had a local recurrence. The rate of incidental second primary malignancies in a specimen was 15.5%.

CONCLUSIONS

A pT0pN0 cystectomy specimen indicates a curative therapy, but there is a substantial risk of tumor recurrence. In the group of tumors with a maximal classification of pT2a, the pT0 tumors constitute a subgroup with a significantly higher likelihood of survival. Cancer 2005. © 2005 American Cancer Society.
Akt activation correlates with adverse outcome in tongue cancer
Authors: Massarelli Erminia , Liu Diane D. , Lee J. Jack , El‐Naggar Adel K. , Lo Muzio Lorenzo , Staibano Stefania , De Placido Sabino , Myers Jeffrey N. , Papadimitrakopoulou Vassiliki A.
Published: October 2005

BACKGROUND

Recent data have shown a significant association between phosphorylated‐Akt (p‐Akt) and failure of local disease control by radiation therapy in head and neck squamous carcinoma (HNSCC), and also that Akt activation correlates with histologic progression of HNSCC from premalignant lesions to invasive cancer. This study evaluated the role of Akt in previously untreated preneoplastic lesions of oral cavity and invasive tongue carcinoma on patient outcome and cancer development.

METHODS

PKB/Akt activation was assessed by immunohistochemistry using a phosphorylation state‐specific antibody (Ser 473) in tongue cancer and preneoplastic specimens of oral cavity.

RESULTS

The expression of p‐Akt was detected in 24 (46%) of the 52 available tongue cancer cases and in 10 (45%) of the 22 available preneoplastic lesions. In tongue cancer, with a median follow‐up of 7.3 years, p‐Akt was highly expressed in the cases that relapsed (15 of 17, 88%) or died of cancer (10 of 12, 83%). Disease‐free survival was significantly shorter in cases with Akt expression (log rank test, P < 0.0001) independently of the stage and nodal status.

CONCLUSIONS

Expression of p‐Akt correlated with worse outcome in patients with tongue cancer. This finding highlights the potential role of Akt as a prognostic marker and as a potential target for molecular therapeutics. Cancer 2005. © 2005 American Cancer Society.
Trends in survival from primary central nervous system lymphoma, 1975–1999
Authors: Panageas Katherine S. , Elkin Elena B. , DeAngelis Lisa M. , Ben‐Porat Leah , Abrey Lauren E.
Published: October 2005

BACKGROUND

The age‐adjusted incidence of primary central nervous system lymphoma (PCNSL) has increased since the 1970s, and treatment for this disease has evolved considerably. The objective of this study was to examine time trends in overall survival and disease‐specific mortality in a population‐based cohort of patients with PCNSL.

METHODS

We identified patients diagnosed with PCNSL from 1975–1999 in the Surveillance, Epidemiology, and End Results (SEER) cancer registries. To assess time trends, year of diagnosis was classified in 5‐year intervals: 1975–1980, 1981–1985, 1986–1990, 1991–1995, and 1996–1999. Overall survival distributions were estimated via Kaplan‐Meier methodology and a competing risk analysis was used to assess PCNSL‐specific mortality. We used information on underlying cause of death to distinguish likely immunocompetent patients from those whose PCNSL was related to human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). We also examined survival stratified by age at diagnosis.

RESULTS

From 1975–1999, 2462 patients were diagnosed with PCNSL in SEER. Median survival was 4 months (95% CI 4, 5) for the entire cohort and 9 months (95% CI 8, 11) for the immunocompetent cohort (n = 1565). In the immunocompetent cohort, 965 of 1323 (73%) deaths were attributed to PCNSL. No significant time trend was observed in either overall or PCNSL‐specific survival.

CONCLUSIONS

Overall survival for patients with PCNSL has not improved consistently in the past three decades despite important therapeutic advances during this time. Although results from clinical trials suggest progress in the treatment of PCNSL, survival improvements are not reflected in this population‐based cohort. Cancer 2005. © 2005 American Cancer Society.
Tumour spread in serosal cavities: What have we learned?
Authors: Lærum Ole Didrik
Published: October 2005

During the Montebello Conference on malignant serosal tumours at Lillehammer, Norway, in June 2004, a group of 30 international experts addressed the biologic and genetic aspects of malignant tumours affecting serosal cavities in the human body. Three neoplasms were mainly dealt with: mesotheliomas arising locally, ovarian carcinomas developing in close proximity to the serosa, and breast tumours in which the spread came from some distance. New, important data on the tumour microenvironment and the process of carcinogenesis with progression and acquisition of invasive properties shed new lights on the mechanisms, including proliferative properties, alterations of signal transduction pathways, and tissue remodelling by proteolytic enzymes in the metastasizing cells. Several of these markers have considerable diagnostic and clinical interest. In addition, new aspects of morphologic and immunocytochemical characteristics of the cells as well as genetic markers may soon become powerful tools for practical use. The molecular fingerprint of the individual tumours may also give guidelines for chemotherapy as well as biologic therapies, including induction of apoptosis. The easy accessibility of tumours from serosal fluids and possibilities for specific discrimination of the neoplastic cells from admixed leukocytes and other cells are promising avenues for cytodiagnostics. Diagn. Cytopathol. 2005;33:292–293. © 2005 Wiley‐Liss, Inc.
Differentiation of cardiac Purkinje fibers requires precise spatiotemporal regulation of Nkx2‐5 expression
Authors: Harris Brett S. , Spruill Laura , Edmonson Angela M. , Rackley Mary S. , Benson D. Woodrow , O'Brien Terrence X. , Gourdie Robert G.
Published: October 2005

Nkx2‐5 gene mutations cause cardiac abnormalities, including deficits of function in the atrioventricular conduction system (AVCS). In the chick, Nkx2‐5 is elevated in Purkinje fiber AVCS cells relative to working cardiomyocytes. Here, we show that Nkx2‐5 expression rises to a peak as Purkinje fibers progressively differentiate. To disrupt this pattern, we overexpressed Nkx2‐5 from embryonic day 10, as Purkinje fibers are recruited within developing chick hearts. Overexpression of Nkx2‐5 caused inhibition of slow tonic myosin heavy chain protein (sMHC), a late Purkinje fiber marker but did not affect Cx40 levels. Working cardiomyocytes overexpressing Nkx2‐5 in these hearts ectopically up‐regulated Cx40 but not sMHC. Isolated embryonic cardiomyocytes overexpressing Nkx2‐5 also displayed increased Cx40 and suppressed sMHC. By contrast, overexpression of a human NKX2‐5 mutant did not effect these markers in vivo or in vitro, suggesting one possible mechanism for clinical phenotypes. We conclude that a prerequisite for normal Purkinje fiber maturation is precise regulation of Nkx2‐5 levels. Developmental Dynamics 235:38–49, 2006. © 2005 Wiley‐Liss, Inc.
Notch1 signals through Jagged2 to regulate apoptosis in the apical ectodermal ridge of the developing limb bud
Authors: Francis Jeffrey C. , Radtke Freddy , Logan Malcolm P.O.
Published: October 2005

The Notch family of receptors is involved in a wide variety of developmental processes, including cell fate specification, cell proliferation, and cell survival decisions during cell differentiation and tissue morphogenesis. Notch1 and Notch ligands are expressed in the developing limbs, and Notch signalling has been implicated in the formation of a variety of tissues that comprise the limb, such as the skeleton, musculature, and vasculature. Notch signalling has also been implicated in regulating overall limb size. We have used a conditional allele of Notch1 in combination with two different Cre transgenic lines to delete Notch1 function either in the limb mesenchyme or in the apical ectodermal ridge (AER) and limb ectoderm. We demonstrate that Notch signalling, involving Notch1 and Jagged2, is required to regulate the number of Fgf8‐expressing cells that comprise the AER and that regulation of the levels of fibroblast growth factor signalling is important for the freeing of the digits during normal limb formation. Regulation of the extent of the AER is achieved by Notch signalling positively regulating apoptosis in the AER. We also demonstrate that Notch1 is not required for proper formation of all the derivatives of the limb mesenchyme. Developmental Dynamics 234:1006–1015, 2005. © 2005 Wiley‐Liss, Inc.
Analysis of the regulation of lin‐41 during chick and mouse limb development
Authors: Lancman Joseph J. , Caruccio Nicholas C. , Harfe Brian D. , Pasquinelli Amy E. , Schageman Jeoffrey J. , Pertsemlidis Alexander , Fallon John F.
Published: October 2005

We have cloned the chicken and mouse orthologues of the Caenorhabditis elegans heterochronic gene lin‐41. During limb development, lin‐41 is expressed in three phases over developmental time and most notably is associated with the developing autopod. Using chicken and mouse mutants and bead implantations, we report that lin‐41 is genetically and biochemically downstream of both the Shh and Fgf signaling pathways. In C. elegans, it is proposed that lin‐41 activity is temporally regulated by miRNAs (let‐7 and lin‐4) that bind to complementary sites in the lin‐41 3′‐untranslated region (UTR). Taking a bioinformatics approach, we also report the presence of potential miRNA binding sites in the 3′‐UTR of chicken lin‐41, including sites for the chicken orthologues of both C. elegans let‐7 and lin‐4. Finally, we show that these miRNAs and others are expressed in the chick limb consistent with the hypothesis that they regulate chicken Lin‐41 activity in vivo. Developmental Dynamics 234:948–960, 2005. © 2005 Wiley‐Liss, Inc.
Distinct regulatory elements mediate the dynamic expression pattern of Nkx3.1
Authors: Chen Hui , Mutton Laura N. , Prins Gail S. , Bieberich Charles J.
Published: October 2005

Loss of Nkx3.1 function in mice results in defects in prostate development and epithelial hyperplasia, indicating that this gene plays important roles in both the initiation and maintenance of prostate differentiation. In humans, decreased NKX3.1 expression is associated with the progression of prostate cancer. Despite these roles in prostate development and disease, the transcriptional regulation of Nkx3.1 has not been systematically addressed. A reporter gene approach in transgenic mice was used to identify regulatory regions that dictate the expression pattern of Nkx3.1. A 32‐kb DNA fragment from the Nkx3.1 locus that specifies the expected expression pattern during embryogenesis and postnatal life has been identified. Deletion analyses demonstrated that cis‐regulatory elements that mediate expression in distinct sites are separable. A 5‐kb fragment downstream of the Nkx3.1 coding region contains elements that support expression in the prostate and bulbourethral glands, whereas an upstream fragment contains elements that direct expression in somites and testes. Reporter gene expression analyses also revealed several previously unknown sites of Nkx3.1 expression in males, including urethral glands, glandular cells in the urethral diverticulum and basal epithelial cells in the prostate. In addition, these analyses revealed Nkx3.1 expression in female urethral glands. The identification of Nkx3.1 cis‐regulatory elements provides a unique starting point to dissect signaling pathways involved in prostate organogenesis and pathogenesis and provides a system to perturb gene expression throughout prostate development. Developmental Dynamics 234:961–973, 2005. © 2005 Wiley‐Liss, Inc.
A case of mucormycosis limited to the parotid gland
Authors: Chandu Arun , MacIsaac Richard J. , MacGregor Duncan P. , Campbell Matthew C. , Wilson Michael J. , Bach Leon A.
Published: October 2005

Background.

Mucormycosis is a rare fungal infection commonly affecting structures in the head and neck such as air sinuses, orbits, and the brain. Common predisposing factors include diabetes and immunosuppression. To date, only one case of mucormycosis involving the parotid gland has been reported, and this infection was associated with a fatal outcome.

Methods.

We report a case of parotid gland mucormycosis in a 45‐year‐old woman with type 2 diabetes, who was successfully treated with a superficial parotidectomy and intravenous amphotericin B.

Results.

After initial surgical and antifungal therapy, the patient was left with a residual facial nerve palsy for which multiple sling procedures were performed. She is currently alive and well 6 years after the diagnosis of mucormycosis.

Conclusions.

Mucormycosis of the parotid gland is a rare form of this often‐fatal infection. In this case, infection remained isolated to the parotid gland and was diagnosed soon after presentation. The patient most likely survived because of the early diagnosis, successful surgical removal of all infected tissue, use of intravenous amphotericin therapy, and the aggressive management of comorbidities such as her diabetes. © 2005 Wiley Periodicals, Inc. Head Neck 27: XXX–XXX, 2005
Antifungal and antibacterial properties of a silver‐loaded cellulosic fiber
Authors: Hipler Uta‐Christina , Elsner Peter , Fluhr Joachim W.
Published: October 2005

The skin is the interface between the body and the environment. Each skin type has a specific skin physiology and is more or less adapted for protection against multiple stress factors. Textiles on the other hand are the tissues with the longest contact to the human skin. They play a critical role especially in skin conditions with an increased rate of bacterial and fungal infections like atopic dermatitis and hyperhidrosis, and in diabetic patients and aged skin. The present study demonstrates the antifungal and antibacterial effects of SeaCell® Active in an in vitro test system against Candida albicans (DSM 11225), Candida tropicalis (ATCC 1169), and Candida krusei (ATCC 6258). Furthermore, the antibacterial activity of fibers with different amounts of SeaCell® Active fibers in a dose‐dependent manner against Staphylococcus aureus (ATCC 22923) and Escherichia coli (ATCC 35218) could be demonstrated. If this fiber seems to be suited for bioactive textiles in specific anatomical regions and skin conditions with a susceptibility for fungal and bacterial infections due to Candida species, namely Staphylococcus aureus and Escherichia coli, must be examined by means of further investigations, especially in vivo tests in human, considering allergic and toxic effects of the fiber. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006
Augmentation of implant purchase with bone cements: An in vitro study of injectability and dough distribution
Authors: Gisep A. , Curtis R. , Hänni M. , Suhm N.
Published: October 2005

Vertebroplasty is widely used to treat (augment) osteoporotic fractures of the spine. This technique—with or without metallic implants—might have more widespread indications, if the mechanics of the injection and distribution of the cement dough through cannulated instruments and implants were better understood. This study was performed to investigate injectability of calcium phosphate and acrylic bone cements through implant prototypes, which featured different perforated sleeve designs. Using a custom‐made capillary rheometer, the forces needed to inject 10 mL of the cement dough through standard cannulas were measured in the first series of experiments. In the second series, plastic sleeves were attached to the rheometer, simulating the implant. In both series, the dough was injected into ambient laboratory atmosphere, and in the second series, cement distribution was analyzed by means of an optical system. Injection of cement dough through the cannulas required forces between 50 and 400 N in the case of acrylic cements and between 40 and 500 N in case of the calcium phosphate cements. Using different sleeves did not have a significant influence on the distribution of the cement dough around the sleeve. The amount of cement dough injected was reduced when a perforated implant was attached to the cannula. More material was delivered through the proximal holes of the implant, leading to a V‐shaped distribution of the cement dough. Among topics to be investigated in future studies is determination of the injectability of cement dough into trabecular bone or bone‐like structures. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006
Chemical characteristics and cytocompatibility of collagen‐based scaffold reinforced by chitin fibers for bone tissue engineering
Authors: Li Xiaoming , Feng Qingling , Wang Wenjie , Cui Fuzhai
Published: October 2005

Chitin is a kind of seemly material to match PLLA for a scaffold, which may create an appropriate environment for the regeneration of tissues. In this study, we prepared and evaluated a new nano‐hydroxyapatite/collagen/PLLA (nHACP) scaffold reinforced by chitin fibers for bone‐tissue engineering. The chitin fibers were crosslinked with PLLA by dicyclohexylcarbodimide (DCC). The chemical characteristics were evaluated by Fourier transformed infrared (FTIR) spectroscopy and X‐ray photoelectron spectroscopy (XPS). The mechanical strength was measured by compressive tests. The fibers, crosslinked with PLLA, could enhance the compressive strength of the scaffold about four times. Human marrow mesenchymal stem cells (MSCs) culture showed that the reinforced nHACP scaffolds were more cytocompatible than that without reinforcement. The crosslinks hardly affected the cytocompatibility of the reinforced scaffolds. The results suggested that the reinforced scaffolds (DCC crosslinked) might be a promising candidate for bone‐tissue engineering. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006
A clinical case report: Interface analysis of a successful well‐functioning transmandibular implant from a cadaver mandible
Authors: Nomura Tsutomu , Gold Evan , Powers Michael P. , Shingaki Susumu , Saito Chikara , Katz J. Lawrence
Published: October 2005

Transmandibular implants (TMI) are indicated both for functional reconstruction of the severely atrophic mandible and when routine augmentation is unpredictable. This study investigates the interface of bone around a TMI, retrieved from the cadaver. The TMI had successfully functioned for 7 years. The mandible was immersed in 10% formaldehyde and sectioned into nine appropriate pieces. Samples were embedded in polymethylmethacrylate, and cut around the transosseous posts and cortical screws in both vertical and horizontal sections. Samples were analyzed at 400 MHz (nominal lateral resolution, 2.5 μm) using a UH3 Scanning Acoustic Microscope (Olympus, Tokyo, Japan). The middle of implant specimens 1–4 were cut to 50 μm, and stained by toluidine blue for light microscopy. Dental X‐rays showed no bone resorption around any implant. On a 2‐mm lateral scan, almost uniform interface space was seen between bone and implant surface in cortical screws. There are wider spaces around the transmandibular posts in the superior area. Histology revealed the small area of direct contact. There is bone marrow space in the interface, with no significant fibrous tissue. We interpret these results at the interface to be because of adaptation for stress distribution © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006
New oxidation treatment of NiTi shape memory alloys to obtain Ni‐free surfaces and to improve biocompatibility
Authors: Michiardi A. , Aparicio C. , Planell J.A. , Gil F.J.
Published: October 2005

Various oxidation treatments were applied to nearly equiatomic NiTi alloys so as to form a Ni‐free protective oxide on the surface. Sample surfaces were analyzed by X‐ray Photoelectron Spectroscopy, and NiTi transformation temperatures were determined by differential scanning calorimetry (DSC) before and after the surface treatment. An ion release experiment was carried out up to one month of immersion in SBF for both oxidized and untreated surfaces. The results show that oxidation treatment in a low‐oxygen pressure atmosphere leads to a high surface Ti/Ni ratio, a very low Ni surface concentration and a thick oxide layer. This oxidation treatment does not significantly affect the shape memory properties of the alloy. Moreover, the oxide formed significantly decreases Ni release into exterior medium comparing with untreated surfaces. As a consequence, this new oxidation treatment could be of great interest for biomedical applications, as it could minimize sensitization and allergies and improve biocompatibility and corrosion resistance of NiTi shape memory alloys. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006
Optimal binary covering codes of length 2j
Authors: Weakley William D.
Published: October 2005

The minimum size of a binary covering code of length n and covering radius r is denoted by K(n,r), and codes of this length are called optimal. For j > 0 and n = 2j, it is known that K(n,1) = 2 · K(n−1,1) = 2n − j. Say that two binary words of length n form a duo if the Hamming distance between them is 1 or 2. In this paper, it is shown that each optimal binary covering code of length n = 2j, j > 0, and covering radius 1 is the union of duos in just one way, and that the closed neighborhoods of the duos form a tiling of the set of binary words of length n. Methods of constructing such optimal codes from optimal covering codes of length n − 1 (that is, perfect single‐error‐correcting codes) are discussed. The paper ends with the construction of an optimal covering code of length 16 that does not contain an extension of any optimal covering code of length 15. © 2005 Wiley Periodicals, Inc. J Combin Designs
Extending the Erdős–Ko–Rado theorem
Authors: Tokushige Norihide
Published: October 2005

Let ${\cal F}$ be a k‐uniform hypergraph on n vertices. Suppose that $|F_{1}\cap \cdots \cap F_{r}|\ge t$ holds for all $F_{1},\ldots ,F_{r}\in {\cal F}$. We prove that the size of ${\cal F}$ is at most ${{n-t}\choose {k-t}}$ if $p= {k \over n}$ satisfies

and n is sufficiently large. © 2005 Wiley Periodicals, Inc. J Combin Designs
A classification of all symmetric block designs of order nine with an automorphism of order six
Authors: Crnković Dean , Rukavina Sanja , Schmidt Marcel
Published: October 2005

We complete the classification of all symmetric designs of order nine admitting an automorphism of order six. As a matter of fact, the classification for the parameters (35,17,8), (56,11,2), and (91,10,1) had already been done, and in this paper we present the results for the parameters (36,15,6), (40,13,4), and (45,12,3). We also provide information about the order and the structure of the full automorphism groups of the constructed designs. © 2005 Wiley Periodicals, Inc. J Combin Designs 14: 301–312, 2006
Involvement of TRAIL/TRAIL‐receptors in human intestinal cell differentiation
Authors: Rimondi Erika , Secchiero Paola , Quaroni Andrea , Zerbinati Carlotta , Capitani Silvano , Zauli Giorgio
Published: October 2005

Despite the fact that tumor necrosis factor (TNF)‐related apoptosis inducing ligand (TRAIL) and its receptors (TRAIL‐Rs) are expressed in intestinal mucosa, little is known about the biological role of this system in intestinal cell physiology. The expression of surface TRAIL and TRAIL‐R1, ‐R2, ‐R3, ‐R4 were examined by flow cytometry in the immortalized human cell line tsFHI under culture conditions promoting growth or growth arrest and expression of differentiated traits. A progressive increase of surface TRAIL expression paralleled tsFHI differentiation, consistently with immunohistochemistry analysis showing an increase of TRAIL immunostaining along the crypt–villus axis in normal jejuneal mucosa. In spite of the presence of TRAIL‐R1 and TRAIL‐R2 “death receptors,” recombinant TRAIL was not cytotoxic for tsFHI cells. Exposure of tsFHI to recombinant TRAIL rather increased/anticipated the expression levels of the cyclin‐dependent kinase inhibitors p21 and p27, which mediate the induction of growth arrest and the stabilization of differentiated traits, respectively, as well as of the canonical differentiation marker DPPIV. The differentiation inducing activity of TRAIL was abolished by pre‐incubation with a Fc‐TRAIL‐R2 chimera. On the other hand, TRAIL did not significantly modulate the levels of osteoprotegerin (OPG), CXCL8/IL‐8, CXCL9/MIG, and CXCL10/IP10 spontaneously released or induced by inflammatory cytokines. Taken together, these data suggest that TRAIL might act as a paracrine trophic cytokine on intestinal epithelium, promoting intestinal cell differentiation. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.
Stable knockdown of CFTR establishes a role for the channel in P2Y receptor‐stimulated anion secretion
Authors: Palmer Melissa L. , Lee So Yeong , Carlson Dan , Fahrenkrug Scott , O'Grady Scott M.
Published: October 2005

P2Y receptor regulation of anion secretion was investigated in porcine endometrial gland (PEG) epithelial cells. P2Y2, P2Y4, and P2Y6 receptors were detected in monolayers of PEG cells and immunocytochemistry indicated that P2Y4 receptors were located in the apical membrane. Apical membrane current measurements showed that Ca2+‐dependent and PKC‐dependent Cl− channels were activated following treatment with uridine triphosphate (UTP) (5 µM). Current‐voltage relationships comparing calcium‐dependent and PKC‐dependent UTP responses under biionic conditions showed significant differences in selectivity between Cl− and I− for the PKC‐dependent conductance (PI/PCl = 0.76), but not for Ca2+‐dependent conductance (PI/PCl = 1.02). The I−/Cl− permeability ratio for the PKC‐dependent conductance was identical to that measured for 8‐cpt cAMP. Furthermore, PKC stimulation using phorbol 12‐myristate 13‐acetate (PMA) activated an apical membrane Cl− conductance that was blocked by the CFTR selective inhibitor, CFTRinh‐172. CFTR silencing, accomplished by stable expression of small hairpin RNAs (shRNA), blocked the PKC‐activated conductance associated with UTP stimulation and provided definitive evidence of a role for CFTR in anion secretion. CFTR activation increased the initial magnitude of Cl− secretion, and provided a more sustained secretory response compared to conditions where only Ca2+‐activated Cl− channels were activated by UTP. Measurements of [cAMP]i following UTP and PMA stimulation were not significantly different than untreated controls. Thus, these results demonstrate that UTP and PMA activation of CFTR occurs independently of increases in intracellular cAMP and extend the findings of earlier studies of CFTR regulation by PKC in Xenopus oocytes to a mammalian anion secreting epithelium. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.
TNF‐α‐related apoptosis‐inducing ligand decoy receptor DcR2 is targeted by androgen action in the rat ventral prostate
Authors: Vindrieux David , Réveiller Marie , Florin Anne , Blanchard Cécile , Ruffion Alain , Devonec Marian , Benahmed Mohamed , Grataroli Renée
Published: October 2005

The apoptotic cell death process in the prostate is known to be under the control of androgens. Tumor necrosis factor‐α (TNF‐α)‐related apoptosis‐inducing ligand (TRAIL) is a member of the TNF‐α family of cytokines, known to induce apoptosis upon binding to its death domain‐containing receptors, DR4/TRAIL‐R1 and DR5/TRAIL‐R2. Two additional TRAIL receptors, DcR1/TRAIL‐R3 and DcR2/TRAIL‐R4, lack functional death domains and act as decoy receptors for TRAIL. In this study, we examined whether TRAIL and cellular receptors expression was targeted by androgens during the apoptotic cell death process in the hormone sensitive ventral prostate. The role of androgens was investigated using two sets of experiment. (1) Androgen deprivation associated with an apoptotic process resulted in a decrease in DcR2 mRNA and protein expression in the ventral prostate 3 days after castration. Testosterone administration to castrated adult rats prevented the decrease in DcR2 mRNA and protein levels in the ventral prostate. In contrast, DcR2 expression was modified, neither in the dorsolateral nor in the anterior prostate following castration. No changes were observed in DR4, DR5, DcR1, and TRAIL mRNA and protein levels in prostate after castration. (2) A specific decrease in DcR2 expression was observed in the ventral prostate after treatment of rats with the anti‐androgen flutamide. Together, the present results suggest that testosterone specifically controls DcR2 expression in the adult rat ventral prostate. Androgen withdrawal, by reducing DcR2 expression, might leave the cells vulnerable to cell death signals generated by TRAIL via its functional receptors. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.
Characterisation of serum‐induced intracellular Ca2+ oscillations in primary bone marrow stromal cells
Authors: Foreman Megan A. , Smith Janet , Publicover Stephen J.
Published: October 2005

Intracellular Ca2+ signalling is pivotal to cell function and [Ca2+]i oscillations permit precise and prolonged modulation of an array of Ca2+‐sensitive processes without the need for extended, global elevations in [Ca2+]i. We have studied [Ca2+]i signalling in primary rat marrow stromal cells exposed to foetal calf serum (FCS) constituents at concentrations up to those required to promote growth and differentiation in culture. Spontaneous [Ca2+]i signalling was not observed, but exposure to 1% FCS induced regular, sustained Ca2+ oscillations in 41 ± 3% of cells. Incidence of FCS‐induced oscillations was dose‐dependent, saturating at 0.5%. These oscillations were arrested by disruption of Ca2+ stores with 100 nM–1 µM thapsigargin or discharge of mitochondrial membrane potential and were sensitive to blockade of IP3‐receptors by 50 µM 2‐amino‐ethoxydiphenyl borate (2‐APB) and inhibition of phospholipase C with 5 µM U73122. The oscillations decreased in frequency and amplitude following inhibition of Ca2+ influx with EGTA or La3+ but were poorly sensitive to nifedipine (1–10 µM) and Bay K 8644 (300 nM). The factor(s) responsible for inducing [Ca2+]i oscillations are heat stable, insensitive to disulphide bond reduction with 20 mM dithioerythritol and retained by a 30 kDa molecular weight filter. Serum is routinely present in culture medium at 10%–15% [v/v] and marrow stromal cells maintained under culture conditions exhibited sustained oscillations. This is the first demonstration of agonist‐induced complex Ca2+ signals in marrow stromal cells. We conclude that Ca2+ oscillations occur constantly in these cells in culture and are potentially important regulators of cell proliferation and differentiation. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.
Transcriptomic analysis of an in vitro murine model of ovarian carcinoma: Functional similarity to the human disease and identification of prospective tumoral markers and targets
Authors: Urzúa Ulises , Roby Katherine F. , Gangi Lisa M. , Cherry James M. , Powell John I. , Munroe David J.
Published: October 2005

Ovarian cancer is an aggressive disease of poor prognostic when detected at advanced stage. It is widely accepted that the ovarian surface epithelium plays a central role in disease etiology, but little is known about disease progression at the molecular level. To identify genes involved in ovarian tumorigenesis, we carried out a genome‐wide transcriptomic analysis of six spontaneously transformed mouse ovarian surface epithelial (MOSE) cell lines, an in vitro model for human ovarian carcinoma. Loess normalization followed by statistical analysis with control of multiple testing resulted in 509 differentially expressed genes using an adjusted P‐value ≤0.05 as cut‐off. The top 20 differentially expressed genes included 10 genes (Spp1, Cyp1b1, Btg1, Cfh, Mt1, Mt2, Igfbp5, Gstm1, Gstm2, and Esr1) implicated in various aspects of ovarian carcinomas, and other 3 genes (Gsto1, Lcn7, and Alcam) associated to breast cancer. Upon functional analysis, the majority of alterations affected genes involved in glutathione metabolism and MAPK signaling pathways. Interestingly, over 20% of the aberrantly expressed genes were related to extracellular components, suggestive of potential markers of disease progression. In addition, we identified the genes Pura, Cnn3, Arpc1b, Map4k4, Tgfb1i4, and Crsp2 correlated to in vivo tumorigenic parameters previously reported for these cells. Taken together, our findings support the utility of MOSE cells in studying ovarian cancer biology and as a source of novel diagnostic and therapeutic targets. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.
Involvement of protein phosphatase 2A in the maintenance of E‐cadherin‐mediated cell–cell adhesion through recruitment of IQGAP1
Authors: Takahashi Kazuhide , Nakajima Eri , Suzuki Katsuo
Published: October 2005

Serine/threonine protein phosphatase (PP) 2A regulates many biological processes, however it remains unclear whether PP2A participates in cadherin‐mediated cell–cell adhesion. We show here that the core enzyme of PP2A (PP2A‐AC) is localized in the cell–cell adhesion sites between adjacent cells and associated with the E‐cadherin‐catenins complex in non‐malignant human mammary epithelial (HME) cells at confluence. Treatment of the cells with either okadaic acid (OA), an inhibitor of PP2A, or siRNA for the regulatory subunit A of PP2A (PP2A‐A) caused disruption of cell–cell adhesion and F‐actin assembly, without affecting the complex formation of E‐cadherin with β‐ and α‐catenins. While a small GTPase Rac and its effector IQGAP1 were associated with the E‐cadherin‐catenins complex, either OA or PP2A‐A siRNA concomitantly induced the dissociation of IQGAP1, but not Rac, from the complex and the internalization of E‐cadherin from the cell surface. We therefore propose that PP2A plays a crucial role in the maintenance of cell–cell adhesion through recruitment of IQGAP1 to the Rac‐bound E‐cadherin‐catenins complex. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.
Implication of the hypoxia response element of the vegf promoter in mouse models of retinal and choroidal neovascularization, but not retinal vascular development
Authors: Vinores Stanley A. , Xiao Wei‐Hong , Aslam Sadia , Shen Jikui , Oshima Yuji , Nambu Hiroyuki , Liu Hansheng , Carmeliet Peter , Campochiaro Peter A.
Published: October 2005

Retinal neovascularization (NV) and macular edema, resulting from blood‐retinal barrier (BRB) breakdown, are major causes of visual loss in ischemic retinopathies. Choroidal NV (CNV) occurs in diseases of the retinal pigmented epithelium/Bruch's membrane complex and is another extremely prevalent cause of visual loss. We used mice in which the hypoxia response element (HRE) is deleted from the vascular endothelial growth factor (vegf) promoter (Vegfδ/δ mice) to explore the role of induction of VEGF through the HRE in these disease processes. Compared to wild type (Vegf+/+) mice with oxygen‐induced ischemic retinopathy (OIR) in which vegf mRNA levels were increased and prominent retinal NV and BRB breakdown occurred, Vegfδ/δ littermates with OIR failed to increase vegf mRNA levels in the retina and had significantly less retinal NV and BRB breakdown, but showed prominent dilation of some superficial retinal vessels. Vegf+/δ littermates with ischemic retinopathy developed comparable retinal NV to Vegf+/+ mice, exhibited intermediate levels of BRB breakdown, and did not show vasodilation. In a mouse model of CNV, due to laser‐induced rupture of Bruch's membrane, the area of CNV at Bruch's membrane rupture sites was more than tenfold greater in Vegf+/+ mice than in Vegfδ/δ littermates. In contrast to these dramatic differences in pathologic ocular NV, Vegfδ/δ mice showed subtle differences in retinal vascular development compared to Vegf+/+ mice; it was slightly delayed, but otherwise normal. These data suggest that induction of VEGF through the HRE in its promoter is critical for retinal and CNV, but not for retinal vascular development. J.Cell.Physiol. © 2005 Wiley‐Liss, Inc.