bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional three‐dimensional HepG2 aggregate cultures generated from an ultrasound trap: Comparison with HepG2 spheroids
Authors: Liu Jian , Kuznetsova Larisa A. , Edwards Gareth O. , Xu Jinsheng , Ma Mingwen , Purcell Wendy M. , Jackson Simon K. , Coakley W. Terence
Published: April 2007

Three‐dimensional culture systems are an ideal in vitro model being capable of sustaining cell functionalities in a manner that resembles the in vivo conditions. In the present study, we developed an ultrasound trap‐based technique to rapidly produce HepG2 hepatocarcinoma cell aggregates within 30 min. Enhanced junctional F‐actin was observed at the points of cell–cell contact throughout the aggregates. HepG2 aggregates prepared by the ultrasound trap can be maintained in culture on a P‐HEMA‐coated surface for up to 3 weeks. The cells in these aggregates proliferated during the initial 3 days and cell number was consistent during the following maintenance period. Albumin secretion from these HepG2 aggregates recovered after 3 days of aggregate formation and remained relatively stable for the following 12 days. Cytochrome P450‐1A1 activity was significantly enhanced after 6 days with maximal enzyme activity observed between 9 and 18 days. In addition, comparison experiments demonstrated that HepG2 aggregates generated by the ultrasound trap had comparable functional characterizations with HepG2 spheroids formed by a traditional gyrotatory‐mediated method. Our results suggest that HepG2 aggregate cultures prepared through the ultrasound trap‐based technique may provide a novel approach to produce in vitro models for hepatocyte functional studies. J. Cell. Biochem. 102: 1180–1189, 2007. © 2007 Wiley‐Liss, Inc.
Ultrastructure of the protonephridial system in Neodasys chaetonotoideus (Gastrotricha: Chaetonotida) and in the ground pattern of Gastrotricha
Authors: Kieneke Alexander , Arbizu Pedro Martínez , Ahlrichs Wilko H.
Published: April 2007

The taxon Neodasys has a basal position within Gastrotricha. This makes it very interesting for phylogenetic considerations in this group. To complete the reconstruction of the nephridial system in the stem species of Gastrotricha started earlier, we have studied the whole protonephridial system of Neodasys chaetonotoideus by means of complete sets of ultrathin sections and TEM. In many characters, protonephridia of N. chaetonotoideus resemble those of macrodasyidan gastrotrich species. For example, each of the six protonephridia, arranged in three pairs, consists of three distinct cells that constitute the continuous protonephridial lumen. Especially, the terminal cell of the protonephridia of N. chaetonotoideus shows a striking pattern: The perforation of the filter region is a meandering cleft that is continuous with the seam of the enfolded lumen of that cell. With the results presented here and that of former TEM studies, we give a comprehensive idea of the excretory organs in the ground pattern of Gastrotricha. Moreover, we can elaborate on the hypothesized protonephridial system in the stem species of Bilateria. We suggest that a meandering filtration cleft is a feature of the ground pattern of the Bilateria. J. Morphol., 2007. © 2007 Wiley‐Liss, Inc.
Laccase production by the white‐rot fungus Termitomyces clypeatus
Authors: Bose Shilpi , Mazumder Sharmishtha , Mukherjee Mina
Published: April 2007

Laccase was detected in the culture filtrate of white‐rot fungus Termitomyces clypeatus. The enzyme was found at the late phase of submerged growth in a medium containing glucose or cellulose as the carbon source. The present study indicates that laccase produced by T. clypeatus is an intracellular enzyme, released in the medium due to cell lysis at the end of the growing phase. Laccase produced by T. clypeatus is different from the extracellular polyphenol oxidase of T. albuminosus, also produced at the late phase of growth. This is the first report of laccase production by a Termitomyces sp. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Potential of the adhesion of bacteria isolated from drinking water to materials
Authors: Simões Lúcia Chaves , Simões Manuel , Oliveira Rosário , Vieira Maria João
Published: April 2007

Heterotrophic bacteria (11 genera, 14 species, 25 putative strains) were isolated from drinking water, identified either biochemically or by partial 16s rDNA gene sequencing and their adherence characteristics were determined by two methods: i. thermodynamic prediction of adhesion potential by measuring hydrophobicity (contact angle measurements) and ii. by measuring adherence to eight different substrata (ASI 304 and 316 stainless steel, copper, polyvinyl chloride, polypropylene, polyethylene, silicone and glass). All the test organisms were hydrophilic and inter‐species variation in hydrophobicity occurred only for Comamonas acidovorans. Stainless steel 304 (SS 304), copper, polypropylene (PP), polyethylene (PE) and silicone thermodynamically favoured adhesion for the majority of test strains (>18/25), whilst adhesion was generally less thermodynamically favorable for stainless steel 316 (SS 316), polyvinyl chloride (PVC) and glass. The predictability of thermodynamic adhesion test methods was validated by comparison with 24‐well microtiter plate assays using nine reference strains and three adhesion surfaces (SS 316, PVC and PE). Results for Acinetobacter calcoaceticus, Burkolderia cepacia and Stenotrophomonas maltophilia sp. 2 were congruent between both methods whilst they differed for the other bacteria to at least one material. Only A. calcoaceticus had strongly adherent properties to the three tested surfaces. Strain variation in adhesion ability was detected only for Sphingomonas capsulata. Analysis of adhesion demonstrated that in addition to physicochemical surface properties of bacterium and substratum, biological factors are involved in early adhesion processes, suggesting that reliance on thermodynamic approaches alone may not accurately predict adhesion capacity. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Isolation and characterization of lactic acid bacteria from lakes
Authors: Yanagida Fujitoshi , Chen Yi‐Sheng , Yasaki Masatoshi
Published: April 2007

Lactic acid bacteria (LAB) were isolated from lake‐water samples collected at 7 lakes distributed in Yamanashi prefecture, Japan. Sampling was performed year round. 112 cultures were isolated and divided into classes by phenotype and then into groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified eleven different bacterial groups (A to K), and the results showed that the isolates represented seven genera: Lactococcus, Leuconostoc, Enterococcus, Lactobacillus, Carnobacterium, Streptococcus, and Weissella. Lactococcus lactis subsp. lactis was the most abundant lactic acid bacteria found in these lakes. Furthermore, Lactococcus lactis subsp. lactis was also the most abundant lactic acid bacteria found throughout the year. Seasonal differences, numbers of isolates and the species of lactic acid bacteria were also recorded in this study. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Effect of stress on growth, pigment production and morphology of Monascus sp. in solid cultures
Authors: Babitha Sumathy , Soccol Carlos Ricardo , Pandey Ashok
Published: April 2007

The aim of the current work was to study the influence of stress on pigment production in Monascus purpureus under solid‐state fermentation. Thermal stress was induced by incubating the culture at various high temperatures. For giving osmotic stress, different concentration of NaCl and glycerol were added to the solid substrate prior to autoclaving. Morphological studies were done by light microscopy and scanning electron microscopy. When the cells were exposed to high temperature, high glycerol and salt concentration, significant changes in pigment production and growth were observed. High temperature (>45 °C) induced the production of more yellow pigments. High concentration of NaCl induced conidiation and caused a decrease in fungal biomass (up to 50%) but red pigment production increased from 11.86 OD/gds to 20.14 OD/gds. When subjected to glycerol stress, a significant increase in aerial mycelia was observed when compared with the control conditions. The results attain significance for exploiting the fungal culture of Monascus purpureus LPB 97 for producing colors of choice – red, or yellow, or increasing the yield of red pigments considerably. Thus, these results could well impact the commercial aspect of Monascus pigments for industrial application. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Antimicrobial activity of some Australian plant species from the genus Eremophila
Authors: Ndi Chi P. , Semple Susan J. , Griesser Hans J. , Barton Mary D.
Published: April 2007

Plant species of the genus Eremophila (Myoporaceae) are native to Australia and are known to produce a diverse range of unusual secondary compounds. The purpose of this research was to examine the antimicrobial activity of 72 Eremophila species most of which had not been the subject of any previous pharmacological testing. Organic extracts of Eremophila species were screened for antimicrobial activity against Gram‐positive and Gram‐negative bacteria and yeasts of medical importance. Extracts of a number of Eremophila species showed selective activity against Gram‐positive bacteria with MICs for the most active species in the range of 16 to 62 μg/ml for Streptococcus species, and 62 to 250 μg/ml for standard strains of Staphylococcus aureus. Extracts with the greatest activity against standard strains were tested against 68 clinical isolates of multi‐resistant methicillin‐resistant S. aureus (mMRSA). The majority of the clinical isolates were susceptible to concentrations below 62.5 μg/ml for the extracts of E. drummondii, E. linearis, E. serrulata, E. acrida, E. neglecta, E. virens and a new undescribed species affiliated with E. prolata. The extract of E. virens inhibited growth of all 68 clinical mMRSA isolates at the minimum tested concentration of 31 μg/ml. This study has shown for the first time that a number of different Eremophila species manifest selective antibacterial activity against Gram‐positive organisms which are important causes of human disease. It shows that there are several Eremophila species possessing interesting antibacterial activity besides those that have published traditional use. These may yield novel antibacterial compounds with potential to be used in biomedical applications. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
SERS of plant material
Authors: Zeiri L.
Published: April 2007

Colloidal gold and silver were formed by the spontaneous reduction of metal salts by plant tissue—alfalfa seeds, green tea leaves, carrots and red cabbage. The colloids were analyzed using electron microscopy and spectroscopic tools. The reduction process yielded stable gold colloids, but for silver the colloidal particles were bigger and less stable, tending to form aggregates. The formation of metal colloids enabled surface‐enhanced Raman spectra (SERS) measurements, yielding specific vibrational signatures for the plant components in the proximity of the colloids. The main SERS peaks were attributed to nicotinamide adenine dinucleotide (NAD) and other adenine‐containing materials. Other peaks were assigned to flavins [e.g. flavin adenine dinucleotide (FAD)], chlorophyll, lipids and other biocomponents. Since the SERS spectra did not show any antioxidants common to all four different types of plant tissue, it is proposed that NAD and FAD compounds that play an important role in the respiration process may be involved in the metal reduction process. Copyright © 2007 John Wiley & Sons, Ltd.
Noise reduction in Raman spectra: Finite impulse response filtration versus Savitzky–Golay smoothing
Authors: Člupek Martin , Matějka Pavel , Volka Karel
Published: April 2007

Application of the finite impulse response (FIR) filtration technique for the removal of spectral noise and background broadband deformations from the Raman spectra is tested. Optimal parameters of FIR filters are found and their effectiveness is compared with the Savitzky–Golay (SG) smoothing procedure. The FIR filtration is found to be an effective procedure to treat the whole Raman spectra, but high computing power is needed. Copyright © 2007 John Wiley & Sons, Ltd.
Effect of low level laser treatment after installation of dental titanium implant-immunohistochemical study of vascular endothelial growth factor: an experimental study in rats
Authors: Kim Y-D , Kim S-S , Hwang D-S , Kim G-C , Shin S-H , Kim U-K , Kim J-R , Chung I-K
Published: April 2007

This experiment using an animal experimental model was conducted in order to investigate the effect of low-level laser therapy (LLLT) on the healing of the dental titanium implant. The experimental group received LLLT for a week and the control group did not. Each group consisted of 10 rats. Two rats from the groups were euthenized on the day 1, 3, 7, 14, and 21 of the experiment. The expression of vascular endothelial growth factor (VEGF) was investigated. The expression of VEGF was observed from the initial stage of the installation of the implant for both the experimental and control groups. However, the degree of expression was higher in the experimental group. Bone density after installation of dental titanium implant during osseointegration in the experimental group was higher than the control group. The surface and structure of the titanium implant was not damaged by LLL. From the above results, the expression of VEGF during the osseointegration of the dental titanium implant was observed within bone tissue. The application of the low-level laser influenced the expression of VEGF, and resulted in the expansion of metabolic angiogenesis and increased the activity of bone tissue cells.
Optimierte Fertigungstechnologie bei Audi – Laserscannerschweißen an den Türen des neuen A4
Authors: Brockmann Rüdiger
Published: April 2007

Das konventionelle Schweißen mit Festkörperlasern ist als Fügeverfahren fest im automobilen Karosseriebau etabliert. Die Weiterentwicklung dieses Verfahrens führte zum robotergeführten Laserscannerschweißen, bei dem sich die Bewegung eines Knickarmroboters mit den hochdynamischen Positionierbewegungen eines Laser Scanners verbinden lassen. Audi setzt diese neue hocheffiziente Fertigungstechnik erstmals an den Türen des Audi A4‐Nachfolgers ein.
Modellierung des Laserstrahlschweißens – Ein Weg zum Prozessverständnis
Authors: Berger Peter
Published: April 2007

Das Erlangen von Prozessverständnis ist seit den Anfängen der Lasermaterialbearbeitung eine wichtige Triebfeder für die Entwicklung von Modellen. Beim Laserstrahlschweißen widmen sich – von ganz wenigen Ausnahmen abgesehen – alle in der Literatur zu findenden modellhaften Beschreibungen diesem Ziel. Das Spektrum dieser Modelle reicht von analytischen Beschreibungen bis hin zu komplexen numerischen Simulationen. Allen gemeinsam ist, dass sie einen Baustein zum Verständnis beitragen. Auch heute noch ist der Prozess des Laserstrahlschweißens nicht völlig verstanden und es entstehen neue Modelle. Diese sind überwiegend darauf ausgerichtet, Antworten auf Fragen zu finden, die im Zusammenhang mit der Anwendung von Lasern der neuesten Generation aufkommen.
Fabrication of Micro‐ and Nanoscale Polymer Structures by Soft Lithography and Spin Dewetting
Authors: Ferrell Nicholas , Hansford Derek
Published: April 2007

In this work, the process of spin dewetting of a polymer solution on a topographically patterned PDMS mold was used for fabrication of micro‐ and nanaoscale polymer structures. Spin coating was used to provide a fast and reproducible coating. This simple technique was capable of producing a wide range of polymer feature geometries from a single microfabricated mold. This experimental study looks at the effects of the original mold feature geometry as well as the polymer solution concentration on the resultant microstructures. Polystyrene and poly(propyl methacrylate) were used as model polymers. Features with film thickness ranging from <100 nm to >5 µm were obtained using this technique. The process was also extended to fabrication of nanoscale features.

magnified image
Increased toll‐like receptor 9 expression in cervical neoplasia
Authors: Lee Jeong‐Won , Choi Jung‐Joo , Seo Eun Sung , Kim Mi Jin , Kim Woo Young , Choi Chel Hun , Kim Tae‐Joong , Kim Byoung‐Gie , Song Sang Yong , Bae Duk‐Soo
Published: April 2007

Toll‐like receptors (TLRs) recognize pathogen‐associated molecular patterns (PAMPs) and enable innate immune responses. Although TLR9 has been previously considered to be expressed only in immune cells, there is now increasing evidence that TLR9 expression is present in nonimmune cells as well. In this study, we undertook to determine whether TLR9 expression was associated with disease progression in cervical neoplasia. TLR9 expression was evaluated by immunohistochemistry in 55 formalin‐fixed paraffin‐embedded cervical tissues; nine normal cervical specimens, 10 low‐grade cervical intraepithelial neoplasias (CINs), 12 high‐grade CINs, and 24 invasive squamous cell carcinomas (ISCCs). In addition, TLR9 expression was evaluated, at the RNA level, in fresh frozen cervical carcinoma tissues by real‐time quantitative RT‐PCR. Immunohistochemical staining showed that TLR9 expression was undetectable (55.6%) or weak (44.4%) in normal cervical squamous epithelial tissues, however, variable staining was observed in the basal layer of all normal endocervical glands. TLR9 expression, which was mainly observed as cytoplasmic staining, gradually increased in accordance with the histopathological grade in the following order: low‐grade CIN < high‐grade CIN < ISCC (P < 0.001), and in particular, was moderate to strong in 70% of ISCC cases. Furthermore, real‐time quantitative RT‐PCR revealed that TLR9 expression, in tumors, was significantly enhanced compared to normal cervical tissues (P = 0.012). These results suggest that TLR9 may play a significant role in tumor progression of cervical neoplasia and may represent a useful marker for malignant transformation of cervical squamous cells. © 2007 Wiley‐Liss, Inc.
Inhibition of proprotein convertases: Approaches to block squamous carcinoma development and progression
Authors: de Cicco Ricardo López , Bassi Daniel E. , Benavides Fernando , Conti Claudio J. , Klein‐Szanto Andrés J.P.
Published: April 2007

Most proprotein convertase (PC) inhibitors are compounds that act as competitive inhibitors. All of them contain the general cleavage motif RXK/RR that binds to the PC's active site impairing further interactions with their physiological substrates. The first inhibitors synthesized were the acyl‐peptidyl‐chloromethyl ketones that bind to the PC's active site through its peptidyl group and are able to transverse the plasma membrane due to the acyl moiety. For instance, one of the members of this family that exhibits reduced toxicity and has been widely used as an effective general PCs inhbitor is the derivative decanoyl‐RVKR‐chloromethylketone (CMK). Another approach to PC inhibition is based on proteins that contain either a natural or a bioengineered PC cleavage consensus site. In this context, the bioengineered serpin, alpha‐1‐antitrypsin Portland (alpha 1‐PDX or PDX), proved to be a potent inhibitor of furin, the most studied of the cancer‐related PCs. Both PDX and CMK were able to inhibit invasiveness of squamous cell carcinoma cell lines by blocking activation of cancer‐associated PC substrates such as MT‐MMPs, IGF‐1R, and VEGF‐C. A similar effect was produced by inhibiting PC‐mediated processing using furin prosegment. PDX and CMK have also been assayed in vivo using skin carcinogenesis models. Newer promising small molecules and RNA interference approaches are also being developed to inhibit PCs. © 2007 Wiley‐Liss, Inc.
The use of unsymmetrical indirect covariance NMR methods to obtain the equivalent of HSQC‐NOESY data
Authors: Blinov Kirill A. , Williams Antony J. , Hilton Bruce D. , Irish Patrick A. , Martin Gary E.
Published: April 2007

We have recently demonstrated that unsymmetrical indirect covariance NMR methods can be used to mathematically calculate the equivalent of low sensitivity, hyphenated NMR experiments by combining data from a pair of higher sensitivity experiments. The present report demonstrates the application of this method to the combination of HSQC and NOESY spectra to provide results comparable to HSQC‐NOESY data, albeit with greater sensitivity and with considerably less spectrometer time. Copyright © 2007 John Wiley & Sons, Ltd.
PIAS1 interacts with and represses SOX9 transactivation activity
Authors: Oh Hyun Ju , Kido Tatsuo , Lau Yun‐Fai Chris
Published: April 2007

SOX9 is a transcription factor that harbors a HMG box and plays critical roles in developmental processes including sex determination and chondrogenesis. Currently the regulation of its molecular activity is not well characterized. In the present study, we have identified PIAS1 as a regulator for SOX9 activity. Using the GST pull‐down, co‐immunoprecipitation, and co‐localization methods in tissue culture cells and mouse embryonic tissues, we demonstrated that SOX9 interacts with PIAS1 in vitro and in vivo. PIAS1 enhanced the SUMOylation at lysine 396 of mouse SOX9. Mutant SOX9 with a conversion of lysine 396 to arginine had a distinct nuclear localization from SOX9 with covalently attached SUMO‐1. Effects of SOX9 SUMOylation on its transcriptional activity were examined by reporter assays using Vanin‐1 promoter and Col11a2 enhancer constructs. The lysine 396 to arginine conversion significantly increased the reporter gene activity, while covalent attachment of SUMO‐1 to SOX9 by gene fusion dramatically compromised its transcriptional activity on the reporter gene. Effects of SOX9 interaction with PIAS1 on its transcriptional activity were examined by similar reporter assays. PIAS1 was able to repress both wild type SOX9 and SUMOylation‐deficient SOX9‐K396R, suggesting that SOX9 SUMOylation is not absolutely required for the repression by PIAS1. However, the repression was further enhanced by exogenous SUMO‐1 while SUMO‐ligase‐deficient PIAS1 was not able to repress SOX9 activity. Thus, PIAS1 appears to repress SOX9 activity by at least two SUMO‐ligase dependent mechanisms: (1) the SUMOylation of SOX9 and (2) SUMOylation of unknown factors associated with SOX9 and/or PIAS1. Mol. Reprod. Dev. 74: 1446–1455, 2007. © 2007 Wiley‐Liss, Inc.
A new selection criterion to assess good quality ovine blastocysts after vitrification and to predict their transfer into recipients
Authors: Leoni G.G. , Berlinguer F. , Succu S. , Bebbere D. , Mossa F. , Madeddu M. , Ledda S. , Bogliolo L. , Naitana S.
Published: April 2007

The feasibility to accurately select viable embryos would be valuable for improving pregnancy rates and avoiding futile transfer attempts. The aim of our study was to assess if in vitro‐produced embryo quality could be determined by the timing of blastocoelic cavity re‐expansion after vitrification, warming, and in vitro culture using sheep as a model. Blastocysts were produced in vitro, vitrified/warmed, and cultured in TCM‐199 plus 10% FCS for 72 hr. Embryos were divided into two groups: re‐expanded within 8 hr (A) and from 8 to 16 hr (B) of IVC after warming. Fast re‐expanded blastocysts showed higher in vitro hatching rates and total cell number calculated on the hatched blastocysts compared with slow re‐expanded ones (P < 0.01). Peroxide status evaluation (P < 0.01) and TUNEL test (P < 0.05) revealed a higher number of positive cells in group B compared with group A. The quantitative analysis of protein synthesis revealed a higher synthesis in fast compared with slow re‐expanded embryos (P < 0.05). Quantitative RT‐PCR showed that 90‐kDa Heat Shock Protein β was more expressed in group A than in group B (P < 0.05), while the quantity of P34cdc2, Cyclin b, Aquaporin 3, Na/K ATPase, and Actin did not differ between the two groups. Pregnancy rates after transfer to synchronized recipients were higher in fast compared to slow re‐expanded blastocysts (P < 0.05). Our results evidenced that timing of blastocoelic cavity re‐expansion after vitrification/warming and in vitro culture can be considered as a reliable index of in vitro produced embryo quality and developmental potential. Mol. Reprod. Dev. 75: 373–382, 2008. © 2007 Wiley‐Liss, Inc.
Deterministic and statistical size effect during shearing of granular layer within a micro‐polar hypoplasticity
Authors: Tejchman J. , Górski J.
Published: April 2007

The paper deals with numerical investigations of a deterministic and statistical size effect in granular bodies during quasi‐static shearing of an infinite layer under plane strain conditions, free dilatancy and constant pressure. For a simulation of the mechanical behaviour of a cohesionless granular material during a monotonous deformation path, a micro‐polar hypoplastic constitutive relation was used which takes into account particle rotations, curvatures, non‐symmetric stresses, couple stresses and the mean grain diameter as a characteristic length. The proposed model captures the essential mechanical features of granular bodies in a wide range of densities and pressures with a single set of constants. In the paper, a deterministic and statistical size effect is analysed. The deterministic calculations were carried out with an uniform distribution of the initial void ratio for four different heights of the granular layer: 5, 50, 500 and 2000 mm. To investigate the statistical size effect, the Monte Carlo method was applied. The random distribution of the initial void ratio was assumed to be spatially correlated. Truncated Gaussian random fields were generated in a granular layer using an original conditional rejection method. The sufficient number of samples was determined by analysing the convergence of the outcomes. In order to reduce the number of realizations without losing the accuracy of the calculations, stratified and Latin hypercube methods were applied. A parametric analysis of these methods was also presented. Some general conclusions were formulated. Copyright © 2007 John Wiley & Sons, Ltd.
Isolation of chlorogenic acids and their derivatives from Stemona japonica by preparative HPLC and evaluation of their anti‐AIV (H5N1) activity in vitro
Authors: Ge Fan , Ke Changqiang , Tang Wei , Yang Xinzhou , Tang Chunping , Qin Guowei , Xu Rensheng , Li Tianxian , Chen Xinwen , Zuo Jianping , Ye Yang
Published: April 2007

Two chlorogenic acids and five chlorogenic acid derivatives were simultaneously separated and purified from Stemona japonica by preparative high‐performance liquid chromatography. Five of the collected compounds were over 95% pure while the other two compounds were over 90% pure. Their structures were elucidated as 3‐O‐feruloylquinic acid (1), 4‐O‐feruloylquinic acid (2), methyl 3‐O‐feruloylquinate (3), methyl 5‐O‐caffeyolquinate (4), methyl 4‐O‐feruloylquinate (5), ethyl 3‐O‐feruloylquinate (6) and the new compound ethyl 4‐O‐feruloylquinate (7) by UV, NMR and ESI‐MS. All compounds were obtained from Stemona species for the first time, however compounds 6 and 7 are believed to be artefacts from the ethanol extraction. The anti‐AIV (H5N1) activities were evaluated by Neutral Red uptake assay. Compounds 3 and 4 exerted moderate inhibitory effect against AIV (H5N1) in vitro. Copyright © 2007 John Wiley & Sons, Ltd.
Prenatal diagnosis of Smith‐Lemli‐Opitz syndrome (SLOS) by DHCR7 mutation analysis
Authors: Waye John S. , Eng Barry , Nowaczyk Małgorzata J. M.
Published: April 2007

Objectives

We review our experience using mutation analysis of the DHCR7 gene for prenatal diagnosis of Smith‐Lemli‐Opitz syndrome (SLOS), an autosomal recessive disorder of endogenous cholesterol biosynthesis caused by deficiency of 7‐dehydrocholesterol reductase (DHCR7).

Methods and Results

Prenatal diagnosis of SLOS was conducted for 21 pregnancies involving 15 families. DNA was isolated directly from amniotic fluid cells or chorionic villus samples (CVS), and the DHCR7 gene was screened for the parental mutations using PCR‐ARMS and nucleotide sequencing. Sixteen of the pregnancies were unaffected and five were affected. There have been no incidences of misdiagnosed pregnancies.

Conclusions

DHCR7 mutation is a rapid and reliable method for prenatal diagnosis of SLOS, and provides an alternative to specialized biochemical tests for elevated 7DHC in amniotic fluid or CVS. Copyright © 2007 John Wiley & Sons, Ltd.
Prenatal thrombosis of the inferior vena cava and the renal veins
Authors: Smorgick Noam , Herman Arie , Wiener Yifat , Halperin Reuvit , Sherman Dan
Published: April 2007

Objective

This manuscript discusses the clinical presentation, diagnosis, pathophysiology and possible etiologies of prenatal thrombosis of the inferior vena cava and renal veins.

Methods

Eleven cases were identified through a Medline search of the English literature. These cases were reviewed, together with a recent case that was treated in our medical center.

Results

Fetal thrombosis in the aforementioned vessels, presented as hydrops fetalis (33%), acute fetal distress (42%), or as an incidental prenatal ultrasound finding (25%). Underlying maternal conditions were present in 58%, and included chronic conditions (hypertension and diabetes mellitus), acute conditions (pyelonephritis and preeclampsia) and thrombophilia. In 42% cases, the prenatal ultrasound scan revealed an enlarged and hyper‐echogenic affected kidney.

Conclusion

Intrauterine fetal venous thrombosis is a rare phenomenon, probably attributed to a multifactorial process. These factors may include inherited thrombophilia as well as prothrombotic clinical conditions. Copyright © 2007 John Wiley & Sons, Ltd.
Cation sitting in aromatic cages: ab initio computational studies on tetramethylammonium–(benzene)n (n=3–4) complexes
Authors: Cheng Jiagao , Gong Zhen , Zhu Weiliang , Tang Yun , Li Weihua , Li Zhong , Jiang Hualiang
Published: April 2007

Quantum chemistry study was performed on interaction between tetramethylammonium (TMA) and aromatic cages by means of the MP2 method to show how TMA sits in an aromatic cage that is composed of benzenes. The MP2 calculations on TMA–(benzene)n complexes demonstrate that the more the benzene molecules in the aromatic cage, the stronger the binding strength between the cage and TMA. In details, the structure of TMA–(benzene)n (n = 1–4) complexes can be easily constructed by superimposing n TMA‐benzene complexes via TMA, and the binding energies of the TMA–(benzene)n complexes are the sum of the n corresponding TMA‐benzene systems. For instance, the distances between the N of TMA and the plane of the benzene ring are 4.238, 4.252, 4.264 ,and 4.276 Å, respectively, for TMA–(benzene)n (n = 1–4) complexes, and the BSSE corrected binding energies at MP2/6‐311++G** level are −8.8, −17.3, −25.8 and −34.3 kcal/mol, respectively, for TMA– (benzene)n (n = 1–4) complexes. Thus, this study provides us useful information on how a cation interacts with an aromatic cage in terms of complex geometry and binding strength. Copyright © 2007 John Wiley & Sons, Ltd.
Dendritic polyamine architectures with lipophilic shells as nanocompartments for polar guest molecules: A comparative study of their transport behavior
Authors: Krämer M. , Kopaczynska M. , Krause S. , Haag R.
Published: April 2007

Dendritic core‐shell architectures were synthesized by simple melt reactions of polyethylenimine (PEI) with different fatty acids. These systems were investigated towards their ability to encapsulate various guest molecules. Parameters, such as the length of the attached alkyl chain, size of the polymer core, concentration of the core‐shell architecture in solution, pH, and nature of the guest molecule were investigated and compared. Guest molecules that bear anionic groups, such as carboxylate, phosphate, sulfonate, or acidic OH groups, as present in phenol units, are readily encapsulated and transferred from the aqueous phase to the organic phase because of multiple anionic–cationic interactions. Hyperbranched polymer architectures exhibit enhanced encapsulation properties when compared with their linear counterparts. In case of the hyperbranched system PEI25C16 amide higher transport capacities were observed at lower concentrations of the polymer, for example, 26 guest molecules at 10−5 mol L−1 versus 143 at 10−6 mol L−1, suggesting formation of larger aggregates. The aggregation behavior of these polar nanocompartments were investigated at different concentrations by AFM showing particle aggregates of typically 250 nm at a concentration of 10−5 M. The individual particle sizes in these aggregates are similar to the particle size at 10−6 M concentration, typically 4.0–5.5 nm (AFM height). This indicates that aggregate formation takes place at concentrations higher than 10−8 M and transport might be mediated by small aggregates rather than unimolecular particles. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 2287–2303, 2007
Butyl acrylate RAFT polymerization in miniemulsion
Authors: Luo Yingwu , Liu Bei , Wang Zuohe , Gao Jin , Li Bogeng
Published: April 2007

Few successes about butyl acrylate (BA) RAFT miniemulsion homopolymerization were reported, even though styrene, methyl methacrylate, and vinyl acetate had been successfully applied in reversible addition fragmentation transfer (RAFT) miniemulsion polymerization. In this article, four types of RAFT agent with various designed R and Z groups [benzyl dithioisobutyrate (BDIB), 1‐phenylethyl phenyldithioacetate (PEPDTA), cumyl dithioisobutyrate (CDIB), benzyl dithiobenzoate] were used to mediate BA miniemulsion polymerization using the conditions (5 wt % hexadance and sodium dodecyl sulfate) effective for styrene and methyl methacrylate systems. All four types of the RAFT agents effectively control over the bulk polymerization. In contrast, only BDIB resulted in a rather narrow molecular weight distribution in the miniemulsion polymerization. A pronounced inhibition and rate retardation were observed in both bulk and miniemulsion polymerizations mediated by CDIB and benzyl dithiobenzoate. When compared with the bulk polymerization, a much longer inhibition period (over eight times) was observed in the CDIB‐mediated miniemulsion polymerization. It was concluded that only the RAFT agent with the primary R group and Z group with less stabilizing ability to the intermediate radicals is effective to mediate BA miniemulsion polymerization in terms of achieving a narrow molecular weight distribution and short inhibition period. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 2304–2315, 2007
In situ formation of silver nanoparticles within an amphiphilic graft copolymer film
Authors: Kim Yong Woo , Lee Do Kyoung , Lee Kyung Ju , Min Byoung Ryul , Kim Jong Hak
Published: April 2007

Silver nanoparticles were prepared by UV irradiation from silver salts, such as AgBF4 or AgNO3, when dissolved in an amphiphilic film of poly((oxyethylene)9 methacrylate)‐graft‐poly((dimethyl siloxane)n methacrylate), POEM‐g‐mPDMS. The in situ formation of silver nanoparticles in the graft copolymer film was confirmed by transmission electron microscopy (TEM), UV‐visible spectroscopy, and wide angle X‐ray scattering (WAXS). The results demonstrated that the use of AgBF4 yielded silver nanoparticles with a smaller size (∼5 nm) and narrower particle distribution when compared with AgNO3. The formation of silver nanoparticles was explained in terms of the interaction strength of the silver ions with the ether oxygens of POEM, as revealed by differential scanning calorimetry (DSC) and X‐ray photoelectron spectroscopy (XPS). It was thus concluded that a stronger interaction of silver ions with the ether oxygens results in a more stable formation of silver nanoparticles, which produces uniform and small‐sized nanoparticles. DSC and small angle X‐ray scattering (SAXS) data also showed the selective incorporation and in situ reduction of the silver ions within the hydrophilic POEM domains. Excellent mechanical properties of the nanocomposite films (3–5 × 105 dyn/cm2) were observed, mostly because of the confinement of silver nanoparticles in the POEM chains as well as interfaces created by the microphase separation of the graft copolymer film. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 1283–1290, 2007
ErbB‐4 may control behavior of prostate cancer cells and serve as a target for molecular therapy
Authors: Ben‐Yosef Rami , Starr Alex , Karaush Victoria , Loew Vered , Lev‐Ari Shahar , Barnea Itay , Lidawi Ghalib , Shtabsky Alexander , Greif Yoel , Yarden Yosef , Vexler Akiva
Published: April 2007

PURPOSE

To assess ErbB‐4 expression in advanced human prostate cancer (PC) cell lines, the role of ErbB‐4 in motility, migration, and proliferative/tumorigenic potential of PC cells, and efficacy of anti‐ErbB‐4 monoclonal antibody (Mab) treatment on PC cells in vitro and tumor growth in vivo.

MATERIALS AND METHODS

Established advanced human PC cell lines (PC‐3, Cl‐1, and Du‐145) were evaluated for ErbB‐4 expression. Several Cl‐1 cell line clones expressing various levels of ErbB‐4 were isolated, their motility, migration capacity, and in vitro proliferation as well as survival following Mab treatment were evaluated. Tumorigenicity and proliferation capacity of these clones in vivo and efficacy of Mab treatment on tumor growth were estimated by measurements of subcutaneous tumors developed in nude mice.

RESULTS

PC cell lines studied express ErbB‐4. Both PC‐3 and Du‐145 cell lines express high ErbB‐4 levels; only 50% of Cl‐1 cells express ErbB‐4 with large heterogeneity. Cl‐1 sub‐clones highly expressing ErbB‐4 showed increased cell motility, migration, and proliferation rate in vitro and enhanced growth in vivo, compared to clones with low ErbB‐4 expression. Mab treatment inhibited the growth of cells expressing high but not low ErbB‐4 levels in vitro and decreased the growth of subcutaneous tumors in nude mice generated by ErbB‐4 highly expressing cells.

CONCLUSIONS

High expression of ErbB‐4 in prostate cancer Cl‐1 cell clones correlated with high proliferative and migration capacity and high tumorigenic potential. The inhibitory effect of Mab on cell proliferation and on subcutaneous tumor growth suggests ErbB‐4′s potential as a target for molecular anticancer therapy. Prostate 67: 871–880, 2007. © 2007 Wiley‐Liss, Inc.
Analysis of the MDM2 antagonist nutlin‐3 in human prostate cancer cells
Authors: Logan Ian R. , McNeill Hesta V. , Cook Susan , Lu Xiaohong , Lunec John , Robson Craig N.
Published: April 2007

BACKGROUND

Small molecule MDM2 antagonists including nutlin‐3 have been shown to be effective against a range of cancer cell types and nutlin‐3 can inhibit growth of LNCaP xenografts. We compared the efficacy of nutlin‐3 in three prostate cancer cell types and provide an insight into the mechanism of nutlin‐3.

METHODS

Nutlin‐3 efficacy was measured using proliferation assays, cell cycle analysis, apoptosis assays, quantitative RT‐PCR, and immunoblotting. Chromatin immunoprecipitation (ChIP) assays were also performed.

RESULTS

Nutlin‐3 can specifically inhibit proliferation of LNCaP cells through cell cycle arrest and apoptosis. This coincides with increased levels of the p53‐responsive transcripts p21, PUMA, gadd45, and Mdm2 and recruitment of p53 to chromatin. Nutlin‐3 also reduces androgen receptor levels, resulting in altered receptor recruitment to chromatin.

CONCLUSION

Our study demonstrates that small molecule MDM2 antagonists might be useful in the treatment of human prostate cancers that retain functional p53 and androgen receptor signaling. Prostate 67: 900–906, 2007. © 2007 Wiley‐Liss, Inc.
Regulation of prostate cancer cell growth and PSA expression by angiotensin II receptor blocker with peroxisome proliferator‐activated receptor gamma ligand like action
Authors: Ishiguro Hitoshi , Ishiguro Yukari , Kubota Yoshinobu , Uemura Hiroji
Published: April 2007

BACKGROUND

We previously reported that angiotensin II (AII) activated the proliferation of prostate cancer cells, and its antagonist, an AII receptor type 1 (AT1R) blocker (ARB), inhibited the proliferation of prostate cancer in vitro and in vivo. In the present study, we investigated whether telmisartan, an ARB, has a unique feature as a peroxisome proliferator‐activated receptor γ (PPARγ) ligand, and its suppressive potential on prostate cancer cells.

METHODS

Cell count or MTT assay were carried out for growth suppression of prostate cancer cells. Phosphorylation of mitogen‐activated protein kinase (MAPK), specific expression of prostate specific antigen (PSA) and AT1R were investigated by western blot. To confirm the PPARγ activity of ARBs, luciferase assay using PSA promoter and PPARγ response elements (PPRE) plasmids was performed.

RESULTS

The results showed that cell proliferation and signal transduction were inhibited by telmisartan treatment. Also, inhibition of PSA expression by telmisartan was confirmed by western blot and luciferase assay, indicating that an ARB acted in a similar way such as an anti‐androgenic agent in prostate cancer cells.

CONCLUSION

The present study showed ARBs, especially those possessing a PPARγ ligand‐like structure, have a potential antagonistic effect on androgen‐dependent and ‐independent prostate cancer. Prostate 67: 924–932, 2007. © 2007 Wiley‐Liss, Inc.
Immunological evaluation of neoadjuvant peptide vaccination before radical prostatectomy for patients with localized prostate cancer
Authors: Noguchi Masanori , Yao Akihisa , Harada Mamoru , Nakashima Osamu , Komohara Yoshihiro , Yamada Satoko , Itoh Kyogo , Matsuoka Kei
Published: April 2007

BACKGROUND

The purpose of this study was to determine the safety and immune responses of pre‐operative personalized peptide vaccine for patients with localized prostate cancer.

METHOD

Ten human leukocyte antigen (HLA)‐A24+ patients with localized prostate cancer received weekly personalized peptide vaccine for six times with positive peptides (up to four kinds of peptides) from 16 kinds of vaccine candidates, followed by a retropubic radical prostatectomy (RRP). Eight patients with localized prostate cancer receiving RRP served as the control group. The serum prostate‐specific antigen (PSA) level, and peptide‐specific cytotoxic T lymphocyte (CTL) precursor analysis by interferon‐γ production, and peptide‐reactive immunoglobulin G (IgG) using an enzyme‐linked immunosorbent assay were monitored during the treatment. Distributions of CD45RO+ cells, CD8+ T cells, and CD20+ B cells in tissue microarray samples were studied using an immunohistochemical technique.

RESULT

The peptide vaccination was safe and well tolerated with no major adverse effects. Increased CTL response and the anti‐peptide IgG titer were observed in the post‐vaccination samples in 8 of 10 or 8 of 10 patients, respectively. The intensity of CD45RO+ infiltrating cells in the vaccination group was significantly larger than that in the control group. CD8+ T cell infiltration was seen only in the vaccinated group.

CONCLUSION

Increased immune responses, at both the circulation and tumor sites in the vaccinated group, support the further development of personalized peptide vaccines for patients with localized prostate cancer. Prostate 67: 933–942, 2007. © 2007 Wiley‐Liss, Inc.
Claudin‐1 immunohistochemistry for distinguishing malignant from benign epithelial lesions of prostate
Authors: Krajewska Maryla , Olson Allen H. , Mercola Dan , Reed John C. , Krajewski Stan
Published: April 2007

BACKGROUND

Claudins are a family of approximately 23 integral membrane tight junction (TJ) proteins that maintain cell polarity and paracellular barrier functions in epithelial and endothelial cells. Although Claudin‐1 was demonstrated to be typically downregulated in various cancers, the precise expression patterns of this protein in normal and neoplastic tissues remain poorly characterized.

METHODS

Using immunohistochemistry, the expression of Claudin‐1 was investigated in prostate tissue samples arranged in a tissue microarray (TMA) format and comprising elements of normal prostatic epithelium (n = 6), benign prostatic hyperplasia (BPH; n = 38), prostatic intraepithelial neoplasia (PIN; n = 11), and prostate adenocarcinoma (n = 48). The Claudin‐1 expression pattern was compared with that of the basal cell‐specific markers, p63, and HMW cytokeratin (34βE12), by employing double‐labeling techniques in conjunction with image analysis methods utilizing color deconvolution algorithms.

RESULTS

In benign prostatic epithelium, pronounced Claudin‐1 expression was observed in the basal cell layer with no staining in luminal cells. Prostate adenocarcinoma specimens from 98% (47/48) patients lacked Claudin‐1 immunostaining, and no cases contained >5% immunopositive tumor cells.

CONCLUSIONS

Claudin‐1 immunohistochemistry should be considered for use as a new diagnostic tool for distinguishing malignant from benign lesions of the prostate. Prostate 67: 907–910, 2007. © 2007 Wiley‐Liss, Inc.
Labeling and identification of LNCaP cell surface proteins: A pilot study
Authors: Whitaker Hayley C. , Stanbury David P.B. , Brinham Claire , Girling Joanne , Hanrahan Sarah , Totty Nick , Neal David E.
Published: April 2007

BACKGROUND

Membrane proteins provide the interface between the cell and its environment and are responsible for cell adhesion, mobility, and intracellular signaling. Previous studies have focused on the LNCaP whole cell proteome and transcriptome but little is known about proteins at the prostate cell membrane and how they change in response to androgens.

MATERIALS AND METHODS

Following treatment with R1881 or vehicle, membrane proteins of the prostate cancer LNCaP cell line were tagged with biotin using EZ‐link sulfo‐NHS‐LC‐biotin. Using the tag membrane proteins were purified and separated using two‐dimensional gel electrophoresis and identified using mass spectrometry. E‐cadherin and low density lipoprotein receptor (LDLR) were used as positive controls and also investigated following bicalutamide treatment. Membrane localization and androgen‐regulation of proteins was confirmed using sub‐cellular fractionation, Western blotting and microscopy.

RESULTS

We have demonstrated efficient and specific protein biotinylation and purification of LNCaP plasma membrane proteins using Western analysis. E‐cadherin and LDLR were regulated at the cell surface in response to R1881 and bicalutamide. Mass spectrometry identified several androgen‐regulated membrane associated proteins including Prx‐3 and GRP78 which are known to localize to other cellular compartments as well as the plasma membrane. We confirmed the localization of the identified proteins in LNCaP cells by co‐localization with E‐cadherin and immunohistochemistry of prostate tissue.

CONCLUSION

Cell surface biotinylation is an effective technique for identifying membrane proteins in the LNCaP prostate cancer cell line. We have demonstrated the identification of androgen‐regulated membrane proteins and their validation in tissue samples. Prostate 67: 943–954, 2007. © 2007 Wiley‐Liss, Inc.