bluuebunny/crossref_metadata_2025 huggingface dataset:
Protein expression and amplification of AIB1 in human urothelial carcinoma of the bladder and overexpression of AIB1 is a new independent prognostic marker of patient survival
Authors: Luo Jun‐Hang , Xie Dan , Liu Meng‐Zhong , Chen Wei , Liu Yong‐Dong , Wu Guo‐Qing , Kung Hsiang‐Fu , Zeng Yi‐Xin , Guan Xin‐Yuan
Published: February 2008

AIB1, a candidate oncogene in human breast cancer, is frequently amplified and overexpressed in several types of human cancers, but the status of AIB1 amplification and expression in urothelial carcinoma of the bladder (UC) and its clinical/prognostic significance is unclear. In our study, the methods of immunohistochemistry and fluorescence in situ hybridization were utilized to examine protein expression and amplification of AIB1 in 163 primary UCs and in 30 samples of normal bladder mucosa. Overexpression of AIB1 and amplification of AIB1 was found in 32.5 and 7.0% of UCs, respectively. In univariate survival analysis of the UC cohorts, a highly significant association of overexpression of AIB1 with shortened patient survival (mean: 45.6 months vs. 59.0 months, p < 0.001, log rank test) was demonstrated. In different subsets of UC patients, overexpression of AIB1 was also a prognostic indicator in grade 1 (p = 0.007), grade 2 (p = 0.010) and grade 3 (p = 0.015) tumor patients, and in pTa (p = 0.025), pT2–4 (p = 0.004), pN0 (p < 0.001) and pT2–4/pN0 (p = 0.040) tumor patients. Importantly, AIB1 expression (p < 0.001) together with pT and pN status (p < 0.05) provided significant independent prognostic parameters in multivariate analysis. In addition, a significant correlation (p < 0.05) of overexpression of AIB1 with an increased UC labeling index of Ki‐67 (a cell proliferation marker) was observed in these UCs. Thus, these findings provide evidence that an overexpression of AIB1, as detected by immunohistochemistry, is an independent molecular marker for poor prognosis (shortened survival time) of patients with UC. © 2008 Wiley‐Liss, Inc.
High expression of Ran GTPase is associated with local invasion and metastasis of human clear cell renal cell carcinoma
Authors: Abe Hideyuki , Kamai Takao , Shirataki Hiromichi , Oyama Tetsunari , Arai Kyoko , Yoshida Ken‐ichiro
Published: February 2008

The Ran small GTPase (Ran) is involved in the regulation of nuclear transport, microtubule nucleation and dynamics, and spindle assembly. To address the question of whether Ran protein is associated with the progression of renal cell carcinoma (RCC), we compared by Western blotting the Ran protein levels in surgical RCC specimens from 180 consecutive Japanese patients with those in the corresponding nontumor tissue from the same patient. We also examined the Ran protein levels in tumors of different grades and stages. Ran proteins were more abundant in RCC tumor tissues than in nontumor tissues (p < 0.0001). High Ran expression was associated with higher grade, local invasion, and metastasis (p < 0.0001). Kaplan‐Meier plots linked high Ran protein expression to a shorter overall survival in all cases (p < 0.0001) and a shorter disease‐free survival in those without metastasis at radical or partial nephrectomy (M0; 131 cases, p < 0.0001). Ran protein expression was an independent factor influencing overall survival univariate analysis (p < 0.0001) and disease‐free survival by multivariate analysis (p < 0.05). Our findings suggest that Ran is associated with the progression of RCC. © 2008 Wiley‐Liss, Inc.
Breast cancer‐associated mutations in metalloprotease disintegrin ADAM12 interfere with the intracellular trafficking and processing of the protein
Authors: Dyczynska Emilia , Syta Emilia , Sun Danqiong , Zolkiewska Anna
Published: February 2008

ADAM12 has recently emerged as a Candidate Cancer Gene in a comprehensive genetic analysis of human breast cancers. Three somatic mutations in ADAM12 were observed at significant frequencies in breast cancers: D301H, G479E and L792F. The first 2 of these mutations involve highly conserved residues in ADAM12, and our computational sequence analysis confirms that they may be cancer‐related. We show that the corresponding mutations in mouse ADAM12 inhibit the proteolytic processing and activation of ADAM12 in NIH3T3, COS‐7, CHO‐K1 cells and in MCF‐7 breast cancer cells. The D/H and G/E ADAM12 mutants exert a dominant‐negative effect on the processing of the wild‐type ADAM12. Immunofluorescence analysis and cell surface biotinylation experiments demonstrate that the D/H and G/E mutants are retained inside the cell and are not transported to the cell surface. Consequently, the D/H and G/E mutants, unlike the wild‐type ADAM12, are not capable of shedding Delta‐like l, a ligand for Notch receptor, at the cell surface, or of stimulating cell migration. Our results suggest that the breast cancer‐associated mutations interfere with the intracellular trafficking of ADAM12 and result in loss of the functional ADAM12 at the cell surface. © 2008 Wiley‐Liss, Inc.
Assessing the Ecological Integrity of a Major Transboundary Mediterranean River Based on Environmental Habitat Variables and Benthic Macroinvertebrates (Aoos‐Vjose River, Greece‐Albania)
Authors: Chatzinikolaou Yorgos , Dakos Vasilis , Lazaridou Maria
Published: February 2008

Ecological integrity has become a primary objective in monitoring programs of surface waters according to the European Water Framework Directive. For this reason we propose a scheme for assessing the ecological integrity of a major transboundary river, the Aoos‐Vjose (Greece‐Albania), by analysing the effects of physicochemical, hydromorphological and habitat structure variables on benthic macroinvertebrates. Benthos and water samples were obtained from 17 sites, during high and low flow season. Physical habitat structure was determined using the River Habitat Survey method. In all but one of the surveyed habitats no anthropogenic change was evident. Macrobenthos assemblages were mainly influenced by seasonality and river section, whereas the water quality index was negatively correlated to habitat modification. Consequently, a large part of the river is considered of high ecological integrity and as such it may be used as baseline information for the management of other major rivers in the eastern Mediterranean basin. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Seven Decades of Change in the Zooplankton (s.l.) of the Nile Delta Lakes (Egypt), with Particular Reference to Lake Borullus
Authors: Dumont Henri J. , El Shabrawy Gamal M.
Published: February 2008

Around the 1930s, the zooplankton (and benthos) of the Nile delta lakes, and Lake Borullus in particular, had a mixed, eutrophic facies, with marine and mesohaline elements dominant for about eight months per year, and freshwater species taking over during the four months of the Nile flood. After the Aswan dam became operational, this regime changed: a steady supply of agricultural drainage water of Nilotic origin consistently freshened the delta. Thus, except in the immediate vicinity of their outlet to the sea, the lakes became almost fresh. Only during the rare and short‐lived (one‐three weeks) occasions when Aswan closes in winter, marine water is sucked in, and along with it, a saline fauna temporarily becomes re‐established in the east and centre of lake Borullus, and presumably of the other delta lakes as well. This marine fauna remained the same over 70+ years of observations. The freshwater component, in contrast, partly nilotic, partly mediterranean, changed deeply over time. First, the fraction of species from temporary waters disappeared, as well as (among copepods and cladocerans) all large‐bodied species. Several cladocerans and copepods with a euro‐mediterranean range appeared and diluted the pre‐existing Afrotropical fauna. The abundance of small cladocerans and, especially, rotifers increased by a factor ten or more. This latter change is believed to reflect two pressures. In a first phase, a re‐arrangement of the lake's fish fauna (a top down force) occurred. Freshwater fish replaced marine diadromic species, and their predation pressure on the zooplankton preferentially removed large‐bodied prey. In a second phase, increased agricultural drainage caused eutrophication (a bottom‐up force) and larger filtrators (cladocerans, some copepods) began to be replaced by small filtrators (rotifers). (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
The Relative Role of Interference Competition in Regulation of a Rotifer Community during Spring Development in a Eutrophic Reservoir
Authors: Devetter Miloslav , Seďa Jaromír
Published: February 2008

The relative importance of interference and exploitative competition from Daphnia was tested in contrast to known role of Cyclops vicinus predation, influencing the rotifer community in Římov Reservoir during spring period. The abundances of five dominant rotifer species and their birth and death rates were analysed, together with changes in chl‐a concentration, HNF, bacteria biomass and the abundances of predator and competitor species, from high frequency sampling. Development of loricated species (Keratella cochlearis, K. quadrata and Kellicottia longispina) was driven predominantly by food availability. The dominant soft‐bodied species Polyarthra spp. and Synchaeta lakowitziana were affected predominantly through C. vicinus predation. The role of mechanical interference from D. galeata, although may be important in a limited periods was overclouded by predation and exploitative competition in general. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Comparison of Fish Assemblage Diversity in Natural and Artificial Rip‐Rap Habitats in the Littoral Zone of a Large River (River Danube, Hungary)
Authors: Erős Tibor , Tóth Balázs , Sevcsik András , Schmera Dénes
Published: February 2008

We studied day‐night patterns in fish diversity in natural, gravel‐sand stretches and boulder covered rip‐rap habitats in the littoral zone of the River Danube. Sample‐based rarefaction indicated marked differences in species richness between day and night, and smaller differences between habitats for both day and night. Whereas, individual‐based rarefaction indicated no such substantial differences in species richness. However, distinct fish assemblages were found based on relative abundance data, and species of great conservation concern tended to link to natural habitats. The diversity of biological traits/attributes were generally higher in rip‐rap habitats. The differences in fish assemblage characteristics between habitats revealed the importance of gravel‐sand habitats in maintaining natural assemblages and that rip‐rap sections increase compositional beta diversity at the mesohabitat scale. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Significance and Interspecific Variability of Accumulated Trace Metal Concentrations in Antarctic Benthic Crustaceans
Authors: Keil Steffen , De Broyer Claude , Zauke Gerd‐Peter
Published: February 2008

Trace metals (Cd, Cu, Pb and Zn) were analysed in crustaceans collected on Polarstern cruises ANT XVI/2 (1999) and ANT XXI/2 (2003/04, BENDEX) to the Weddell Sea. Our study provides further evidence for the frequently reported “Cd anomaly” in polar crustaceans, with data ranging from 1.2 (Ceratoserolis trilobitoides) to 6.2 mg Cd kg–1 DW (Notocrangon antarcticus) in 1999 and from 1.2 (Waldeckia obesa) to 20.3 mg Cd kg–1 (Tryphosella murrayi) in 2003. Pb concentrations well below 1 mg kg–1 in most of the samples analysed might serve as a regional or even global background value for comparison in biomonitoring studies. Increasing Cu concentrations from eggs of decapods (e.g., 5 vs. 51 mg kg–1 in N. antarcticus) or juveniles in the brood pouch of an amphipod species to adult females indicate that the enzymatic requirements and haemocyanin component demand for Cu in early life‐history stages is probably not met without a distinct bioaccumulation of this essential element after hatching. Most interestingly, Cd also increases (<0.1 vs. 6.2 mg kg–1 in N. antarcticus). This could be the consequence of efficient uptake mechanisms for Cu that cannot discriminate between this element and Cd. Cu and Zn concentrations in decapods of this study are largely within the range reported worldwide (40–90 mg Cu kg–1 and 40–80 mg Zn kg–1), indicating that these elements are regulated. The enormous heterogeneity of Cd and Zn in many amphipod species investigated (e.g., from 0.6 in Gnathiphimedia mandibularis to 34.4 mg Cd kg–1 in Orchomenopsis acanthura and from 41 in Eusirus antarcticus to 1244 mg Zn kg–1 in Iphimediella bransfieldi) supports the hypothesis of the “Cd anomaly” and suggests that there is probably no consistent metabolic demand for the essential element Zn in this taxonomic group. The heterogeneity of Cu in amphipods is less pronounced. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Blocking of intracellular ROS production by phytoglycoprotein (30 kDa) causes anti‐proliferation in bisphenol A‐stimulated Chang liver cells
Authors: Oh Phil‐Sun , Lim Kye‐Taek
Published: February 2008

Dioscorea batatas Decne (DBD) is traditionally used to heal inflammatory disease as a folk medicine. It was reported that a glycoprotein (DBD glycoprotein) with a molecular weight of 30 kDa was isolated from DBD and consists of carbohydrate (83.75%) and protein (16.25%) moieties. The previous results showed that it has a strong scavenging activity against hydroxyl radicals without any pro‐oxidant activity in the cell‐free system. The purpose of the present study was to show whether or not the DBD glycoprotein inhibits cell proliferation‐related signal transduction stimulated by bisphenol A (BPA, an environmental hormone) in Chang liver cells. The results in this study indicated that DBD glycoprotein (200 µg ml−1) has suppressive effects on abnormal cell viability, production of intracellular reactive oxygen species (ROS) and nitric oxide (NO) in BPA (50 µm)‐induced Chang liver cells by blocking the phosphorylation of mitogen‐activated protein kinase (MAPK) and activating protein‐1 (AP‐1) activity. In addition, DBD glycoprotein (200 µg ml−1) normalized the activity of catalase (CAT) and glutathione peroxidase (GPx). Consequently, DBD glycoprotein inhibits the expression of proliferating cell nuclear antigen (PCNA, cell proliferation maker) stimulated by BPA. Therefore, it is speculated that DBD glycoprotein protects against carcinogenic events caused by BPA in Chang liver cells. Copyright © 2008 John Wiley & Sons, Ltd.
Synthesis of a [18F]fluorobenzothiazole as potential amyloid imaging agent
Authors: Berndt Ursula , Stanetty Christian , Wanek Thomas , Kuntner Claudia , Stanek Johann , Berger Michael , Bauer Martin , Henriksen Gjermund , Wester Hans‐Jürgen , Kvaternik Herbert , Angelberger Peter , Noe Christian
Published: February 2008

This study describes the synthesis of a fluoroethylated derivative of [N‐methyl‐11C]2‐(4′‐methylaminophenyl)‐6‐hydroxybenzothiazole ([11C]6‐OH‐BTA‐1; Pittsburgh Compound B (PIB)), an already established amyloid imaging agent. The [11C]methylamino group of [11C]6‐OH‐BTA‐1 was formally replaced by a fluoroethyl group in a cold synthesis via N‐alkylation of N‐Boc‐2‐(4′‐aminophenyl)‐6‐(methoxyethoxymethoxy)benzothiazole with fluoroethyl tosylate. Subsequent deprotection gave the target compound 2‐[4′‐(2‐fluoroethyl)aminophenyl]‐6‐hydroxybenzothiazole (FBTA). In a radioligand competition assay on aggregated synthetic amyloid fibrils using N‐[3H‐methyl]6‐OH‐BTA‐1, 100 nM FBTA inhibited binding with 93 ± 1 and 83 ± 1% efficiency for Aβ1–40 and Aβ1–42, respectively. For the radiosynthesis a precursor carrying a tosylethyl moiety was prepared allowing the introduction of [18F]fluoride via nucleophilic substitution with [18F]tetra‐n‐butyl‐ammonium fluoride (TBAF). Subsequent removal of all protecting groups was performed in a one‐pot procedure followed by semi‐preparative HPLC, delivering the target compound [18F]FBTA in good radiochemical yield of 21% on average and radiochemical purity of ⩾98% at EOS. In vitro autoradiography on human postmortem AD brain tissue slices showed intense cortical binding of [18F]FBTA (1 nM), which was displaced in presence of 6‐OH‐BTA‐1 (1 µM). Brain up‐take was evaluated in wild‐type (wt) mice with microPET imaging. Based on these results, [18F]FBTA appears to be a suitable candidate tracer for amyloid imaging in humans. Copyright © 2008 John Wiley & Sons, Ltd.
Synthesis and biological evaluation of carbon‐11‐labeled acyclic and furo[2,3‐d]pyrimidine derivatives of bicyclic nucleoside analogues (BCNAs) for structure–brain uptake relationship study of BCNA tracers
Authors: Chitneni Satish K. , Balzarini Jan , Celen Sofie , Dyubankova Natalia , Verbruggen Alfons M. , Bormans Guy M.
Published: February 2008

We reported earlier on radiolabeled alkoxyphenyl bicyclic nucleoside analogues (BCNAs) as potential positron emission tomography (PET) reporter probes for imaging of varicella zoster virus thymidine kinase (VZV‐tk) gene in vivo. Despite their favorable physicochemical properties, these tracers are not taken up in the brain in mice. In order to probe the role of the deoxyribose sugar moiety in blood‐brain barrier (BBB) penetration of these molecules, we have synthesized and evaluated a carbon‐11‐labeled acyclic bicyclic nucleoside derivative ([11C]‐10) where the 2′‐deoxyribose sugar is replaced with a (2‐hydroxyethoxy)methyl group and [11C]‐12, which has no sugar moiety but a [11C]methyl group on the N‐3 position of the pyrimidine ring. Methylation was achieved on the phenol ([11C]‐10) or the N‐3 position ([11C]‐12) using [11C]methyl triflate (radiosynthesis). The (non‐radioactive) acyclic O‐methyl derivative 10 has rather poor affinity for the enzyme VZV‐TK in vitro (IC50: 430 µM), compared with the moderate affinity of the BCNA‐base N‐methyl derivative 12 (IC50: 79 µM). In normal mice, none of the two tracers ([11C]‐10 or [11C]‐12) showed significant uptake in the brain, suggesting that compounds containing a furo[2,3‐d]pyrimidine system do not cross the BBB. Copyright © 2008 John Wiley & Sons, Ltd.
Immunohistochemical localization of a ubiquitin ligase HRD1 in murine brain
Authors: Omura Tomohiro , Kaneko Masayuki , Tabei Naoki , Okuma Yasunobu , Nomura Yasuyuki
Published: February 2008

HRD1 is an E3 ubiquitin ligase and plays an important role in endoplasmic reticulum‐associated degradation (ERAD). Parkin‐associated endothelin receptor‐like receptor (Pael‐R) is a substrate of the E3 ubiquitin ligase parkin, which has been implicated in ER stress‐induced cell death in dopamine neurons in autosomal recessive juvenile parkinsonism (AR‐JP). Recently, we demonstrated that endogenous HRD1 interacts with Pael‐R, and that HRD1 promotes the degradation of Pael‐R and protects cell death caused by the accumulation of Pael‐R. Another group recently reported that HRD1 suppresses the toxicity of polyglutamine‐expanded huntingtin. However, the topographical localization of HRD1 protein in the brain, especially related to neurodegenerative disease, is unclear. In this study, we used immunohistochemistry to investigate the topographical localization of HRD1 in the brain and demonstrated that HRD1 immunoreactivity was expressed widely in the substantia nigra pars compacta (SNC) containing dopaminergic neurons and was expressed in the cerebral cortex, hippocampus, dentate gyrus, striatum, globus pallidus, and Purkinje cells of the cerebellar cortex. Furthermore, HRD1 immunoreactivity was detected in the neuronal cells but not in the glial cells. These results suggest that HRD1 may play an important role in maintaining higher brain function, including motor function or learning and memory. In addition, HRD1 may have substrates other than Pael‐R that are implicated in neurodegenerative disorders. © 2008 Wiley‐Liss, Inc.
July droughts over Homogeneous Indian Monsoon region and Indian Ocean dipole during El Niño events
Authors: Ihara Chie , Kushnir Yochanan , Cane Mark A.
Published: February 2008

The monthly summer monsoon rainfall over the Homogeneous Indian Monsoon region (HI) that represents most of the variance of all‐India monsoon rainfall is investigated using observational data from 1880 to 2002. Severe droughts in July occur mostly during El Niño events of the boreal summer monsoon season. They occurred frequently in the late 19th to early 20th century, rarely in the middle of the 20th century, and again occasionally occurred after the 1960s.

During El Niño events, severe droughts in July over HI are significantly associated with smaller sea surface temperature‐ (SST) based Indian Ocean Dipole mode index (SSTDMI) in May and July compared to the years without these outstanding events. The evolution of SSTDMI that usually starts around May/June during El Niño events tends to delay when July precipitation over HI is abnormally low. Niño3 values from May through October are not significantly associated with severe droughts over HI in July during El Niño events indicating that the strength of El Niño events is not related to the occurrences of severe droughts in July. Copyright © 2008 Royal Meteorological Society
A pulsing electric field (PEF) increases human chondrocyte proliferation through a transduction pathway involving nitric oxide signaling
Authors: Fitzsimmons Robert J. , Gordon Stephen L. , Kronberg James , Ganey Timothy , Pilla Arthur A.
Published: February 2008

A potential treatment modality for joint pain due to cartilage degradation is electromagnetic fields (EMF) that can be delivered, noninvasively, to chondrocytes buried within cartilage. A pulsed EMF in clinical use for recalcitrant bone fracture healing has been modified to be delivered as a pulsed electric field (PEF) through capacitive coupling. It was the objective of this study to determine whether the PEF signal could have a direct effect on chondrocytes in vitro. This study shows that a 30‐min PEF treatment can increase DNA content of chondrocyte monolayer by approximately 150% at 72 h poststimulus. Studies intended to explore the biological mechanism showed this PEF signal increased nitric oxide measured in culture medium and cGMP measured in cell extract within the 30‐min exposure period. Increasing calcium in the culture media or adding the calcium ionophore A23187, without PEF treatment, also significantly increased short‐term nitric oxide production. The inhibitor W7, which blocks calcium/calmodulin, prevented the PEF‐stimulated increase in both nitric oxide and cGMP. The inhibitor L‐NAME, which blocks nitric oxide synthase, prevented the PEF‐stimulated increase in nitric oxide, cGMP, and DNA content. An inhibitor of guanylate cyclase (LY83583) blocked the PEF‐stimulated increase in cGMP and DNA content. A nitric oxide donor, when present for only 30 min, increased DNA content 72 h later. Taken together, these results suggest the transduction pathway for PEF‐stimulated chondrocyte proliferation involves nitric oxide and the production of nitric oxide may be the result of a cascade that involves calcium, calmodulin, and cGMP production. © 2008 Orthopaedic Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:854–859, 2008
Early inflammatory response of knee ligaments to prolotherapy in a rat model
Authors: Jensen Kristina T. , Rabago David P. , Best Thomas M. , Patterson Jeffrey J. , Vanderby Ray
Published: February 2008

Prolotherapy is an alternative injection‐based therapy for chronic musculoskeletal pain. Three different proliferants, D‐glucose (dextrose), phenol‐glucose‐glycerine (P2G), and sodium morrhuate, used in prolotherapy are hypothesized to strengthen and reorganize chronically injured soft tissue and decrease pain through modulation of the inflammatory process. Our hypothesis is that commonly used prolotherapy solutions will induce inflammation (leukocyte and macrophage infiltration) in medial collateral ligaments (MCLs) compared to needlestick, saline injection, and no‐injection controls. MCLs of 84 Sprague‐ Dawley rats were injected one time at both the tibial and femoral insertions. Immunohistochemistry (IHC) was used to determine the inflammatory response at three locations (tibial and femoral insertions and midsubstance) 6, 24, and 72 h after dextrose injection compared to saline‐ and no‐injection controls and collagenase (positive control) (n = 4). qPCR was used to analyze gene expression 24 h postinjection (n = 4). Sodium morrhuate, P2G, and needlestick control were also investigated after 24 h (n = 4). In general, inflammation (CD43+, ED1+, and ED2+ cells) increased after prolotherapy injection compared to no‐injection control but did not increase consistently compared to saline and needlestick control injections. This response varied by both location and proliferant. Inflammation was observed at 6 and 24 h postinjection but was resolved by 72 h compared to no‐injection controls (p < 0.05). CD43+ leukocytes and ED2+ macrophages increased compared to needlestick and saline‐injection control, respectively, 24 h postinjection (p < 0.05). Prolotherapy injections created an inflammatory response, but this response was variable and overall, not uniformly different from that caused by saline injections or needlestick procedures. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:816–823, 2008
Comparative assessment of soybean meal with high and low glucosinolate rapeseed–mustard cake as protein supplement on performance of growing crossbred calves
Authors: Ravichandiran S , Sharma Kusumakar , Dutta Narayan , Pattanaik Ashok K , Chauhan J S , Agnihotri Abha
Published: February 2008

BACKGROUND: Feeding of high glucosinolate rapeseed–mustard cakes (RMCs) imparts adverse effects on dry matter (DM) intake, health and overall performance of animals. Recently, plant breeding efforts have resulted in many cultivars of RMCs containing low to moderate levels of glucosinolate in India. The feeding value of RMC cultivars with high and low glucosinolate was evaluated relative to commonly used soybean meal as a protein supplement in growing crossbred calves.

RESULTS: Eighteen growing crossbred calves (62.9 ± 3.8 kg body weight) were randomly allocated to three dietary treatments SBM, LG and HG containing soybean meal, low glucosinolate B. napus (15 µmol glucosinolates g−1) and high glucosinolate B. juncea (135 µmol glucosinolates g−1), respectively. Although daily intake of total DM and wheat straw did not differ (P > 0.05) among the dietary treatments, intake (g/kgW0.75) of concentrate moiety decreased quadratically (P < 0.01) with increasing glucosinolate levels in diets. Nutrient digestibility and balances of N, Ca and P by calves were similar (P > 0.05) among dietary treatments. However, average daily gain (g) decreased and feed conversion ratio values increased quadratically (P < 0.05) with increasing glucosinolate levels. Serum metabolic profile and triiodothyronine remained within the normal range; however, thyroxine changed quadratically.

CONCLUSION: The results suggested that while high glucosinolate RMCs may reduce the palatability and consequently growth rate in crossbred calves, SBM can be replaced completely by low glucosinolate rapeseed without compromising their performance. Copyright © 2008 Society of Chemical Industry
Capillary electrophoresis enzyme immunoassay for alpha‐fetoprotein and thyroxine in human serum with electrochemical detection
Authors: Li Xue‐Mei , Zhang Fan , Zhang Shu‐Sheng
Published: February 2008

A novel CE‐based enzyme immunoassay (CE‐EIA) method was developed in o‐aminophenol (OAP)‐H2O2‐horseradish peroxidase (HRP) system and applied to benign liver disease and hyperthyroidism research in the clinical practical field. In the presented method, after the enzyme immunoreaction, the HRP‐labeled antibody or HRP‐labeled antigen catalyzed the enzyme substrate OAP and H2O2. The product of the enzymatic catalysis reaction 2‐aminophenoxazine‐3‐one (AP) was determined using electrochemical detection on a Pt electrode at the outlet of the reaction capillary. Factors influencing the performance, including running buffer concentration, separation, and detection voltage, were investigated to the optimum conditions. Noncompetitive and competitive models were utilized to detect alpha‐fetoprotein (AFP) and thyroxine (T4) in human sera, respectively. The linear ranges and the detection limits (S/N = 3) were from 1.5 to 66.6 ng/mL and 0.48 ng/mL for AFP, and from 1.7 to 260.0 ng/mL and 1.0 ng/mL for T4. The results of this method were linear proportional to those of spectrophotometric ELISA method, giving a good prospect for a new clinical diagnostic instrument.
Stereoselective quantitation of a serine protease inhibitor using LC‐MS/MS at elevated column temperature
Authors: Shen Jim X. , Merka Eric A. , Dreyer Daniel P. , Clement Robert P. , Hayes Roger N.
Published: February 2008

A LC‐MS/MS method using a LC column packed with sub‐2 micron particles and elevated column temperatures was validated for the quantitation of SCH 503034 diastereomers (SCH 534128 and SCH 534129) in human plasma. The method was validated over the concentration range of 2.5 to 1250 ng/mL. Inter‐assay precision, based on percent relative deviation for n = 18 replicate quality controls, was 4.5% for SCH 534128 and 4.9% for SCH 534129. Inter‐assay accuracy based on n = 18 replicate quality controls was ±7.8% for both SCH 534128 and SCH 534129. The method involved the novel application of ion pairing reagents to increase the stereoselectivity of the separation. Temperature, types of ion pairing reagent, and concentration of ion pairing reagent were all found to play significant roles in the resolution of the SCH 534128 and SCH 534129 diastereomers on a LC column packed with sub‐2 micron particles. Specifically, a sensitivity increase of five‐fold was demonstrated by increasing the column temperature. Without sacrificing resolution, the run time was significantly shortened when the column temperature was elevated to 100°C.
Harmonization of biodiesel specifications
Authors: Alleman Teresa L.
Published: February 2008

Worldwide biodiesel production has grown dramatically over the last several years. Biodiesel standards vary across countries and regions, and there is a call for harmonization. For harmonization to become a reality, standards have to be adapted to cover all feedstocks. Additionally, all feedstocks cannot meet all specifications, so harmonization will require standards to either tighten or relax. For harmonization to succeed, the biodiesel market must be expanded with the alignment of test methods and specification limits, not contracted.
Dietary exposure to brominated flame retardants correlates with male blood levels in a selected group of Norwegians with a wide range of seafood consumption
Authors: Knutsen Helle K. , Kvalem Helen E. , Thomsen Cathrine , Frøshaug May , Haugen Margaretha , Becher Georg , Alexander Jan , Meltzer Helle M.
Published: February 2008

This study investigates dietary exposure and serum levels of polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecane (HBCD) in a group of Norwegians (n= 184) with a wide range of seafood consumption (4–455 g/day). Mean dietary exposure to Sum 5 PBDEs (1.5 ng/kg body weight/day) is among the highest reported. Since concentrations in foods were similar to those found elsewhere in Europe, this may be explained by high seafood consumption among Norwegians. Oily fish was the main dietary contributor both to Sum PBDEs and to the considerably lower HBCD intake (0.3 ng/kg body weight/day). Milk products appeared to contribute most to the BDE‐209 intake (1.4 ng/kg body weight/day). BDE‐209 and HBCD exposures are based on few food samples and need to be confirmed. Serum levels (mean Sum 7 PBDEs = 5.2 ng/g lipid) and congener patterns (BDE‐47 > BDE‐153 > BDE‐99) were comparable with other European reports. Correlations between individual congeners were higher for the calculated dietary exposure than for serum levels. Further, significant but weak correlations were found between dietary exposure and serum levels for Sum PBDEs, BDE‐47, and BDE‐28 in males. This indicates that other sources in addition to diet need to be addressed.
Determination of brominated flame retardants and brominated dioxins in fish collected from three regions of Japan
Authors: Ashizuka Yuki , Nakagawa Reiko , Hori Tsuguhide , Yasutake Daisuke , Tobiishi Kazuhiro , Sasaki Kumiko
Published: February 2008

The concentrations of brominated dioxins which are polybrominated dibenzo‐p‐dioxins/polybrominated dibenzofurans (PBDD/DFs) and mono‐bromo polychlorinated dibenzo‐p‐dioxins/dibenzofurans, polybrominated diphenyl ethers (PBDEs) and tetrabromobisphenol A (TBBPA) were investigated in a total of 45 fish samples collected from three regions in Japan. In the brominated dioxins, 1,2,3,4,6,7,8‐heptabromodibenzofuran (HpBDF) was the most abundant congener, and it was found in seven fish samples at 0.10–25.6 pg/g wet weight (ww). The highest concentration of 1,2,3,4,6,7,8‐HpBDF was found in the pike eel. Regarding other congeners, 2,3,7,8‐tetrabromodibenzo‐p‐dioxin was detected in the sea bream at 0.02 pg/g ww, and 2,3,7,8‐tetrabromodibenzofuran was detected in the conger eel at 0.03 pg/g ww. 3‐Bromo‐2,7,8‐trichlorodibenzofuran was detected in the Sardinella zunasi and the conger eel at 0.01 pg/g ww and 0.02 pg/g ww, respectively. Using toxic equivalency factors of chlorinated dioxins, we calculated the PBDD/DFs concentrations of these fish samples at 0.001–0.256 pg TEQ/g ww. PBDEs were detected in all of the fish samples. The concentrations of total PBDEs were 0.01–2.88 ng/g ww. The seerfish and the yellowtail containd PBDEs in high concentrations. The most dominant congener in most of the fish was 2,2′,4,4′‐tetrabromo diphenyl ether. TBBPA was detected in 29 fish samples at 0.01–0.11 ng/g ww. The mean level of TBBPA was about one‐tenth or less of the total level of PBDEs. A good correlation was obtained between total PBDEs and fat content. On the other hand, no correlation was obtained between TBBPA and fat content. The daily intakes from fish were estimated to be 0.58 ng/kg body weight (bw)/day for total PBDEs, 0.03 ng/kg bw/day for TBBPA, and 0.01 pg TEQ/kg bw/day for brominated dioxins in the case assuming that the average bw of a Japanese adult person is 50 kg and that the average fish consumption is 82 g/day. For PBDEs, the provisionally calculated value was much less than the lowest observed adverse effect level value (1 mg/kg bw/day). For brominated dioxins, the daily intake was at a very low level compared with the Japanese daily intake of polychlorinated dioxins from fish. Even if the value of PBDD/DFs is added to the amount of chlorinated dioxin exposure, it was estimated that it is less than the tolerable daily intake (4 pg TEQ/kg bw/day) in Japan.
Brominated flame retardants in fish and shellfish – levels and contribution of fish consumption to dietary exposure of Dutch citizens to HBCD
Authors: van Leeuwen Stefan P. J. , de Boer Jacob
Published: February 2008

In order to determine the contamination with brominated flame retardants (BFR) in fish regularly consumed by Dutch citizens, 44 samples of freshwater fish, marine fish, and shellfish were analyzed for polybrominated diphenyl ethers (PBDE), tetrabromobisphenol‐A (TBBP‐A) and its methylated derivative (me‐TBBP‐A), and hexabromocyclododecane (HBCD), including its α‐, β‐ and γ‐diastereomers. The highest BFR concentrations were found in pike‐perch and eel from the highly industrialized and urbanized rivers Rhine and Meuse. The sum concentrations of BDE 28, 47, 99, 100, 153, 154, 183, 209, and brominated biphenyl (BB) 153 and HBCD (selection based on The European Food Safety Authority monitoring recommendation) ranged from below quantification limits to 17 ng/g wet weight (ww) in marine fish and in freshwater fish from 0.6 ng/g ww in pike‐perch to 380 ng/g ww in eel. The BDE congener profile in all fish and shellfish samples is dominated by BDE 47, followed by BDE 99, except for eel in which BDE 100 is higher than BDE 99. BDE 209 was detected in two mussel samples, most likely due to BDE 209 contaminated particulate matter in their intestines. Total‐HBCD (as determined by GC/electron capture negative ion (ECNI)‐MS) was detected in 22 out of the 44 samples in concentrations between 0.20 ng/g in marine fish and 230 ng/g ww in eel. Three HBCD diastereomers were determined by HPLC/ESI‐MS/MS. α‐HBCD was the prevalent congener in most fish samples, followed by γ‐HBCD. β‐HBCD, TBBP‐A and me‐TBBP‐A were only detected in a few samples and at low concentrations. A considerable difference was found between HBCD results obtained from GC/ECNI‐MS and HPLC/ESI‐MS/MS: the GC/ECNI‐MS results were 4.4 times higher, according to regression analysis. There is hardly any data on human dietary exposure to HBCD available. We have estimated the fish‐related dietary exposure of HBCD for the average Dutch population. The medium bound intake was estimated at 8.3 ng/day for a 70‐kg person (0.12 ng/kg bodyweight/day). For this estimation, we relied mostly on HPLC/ESI‐MS/MS data as we argue that these results are more accurate than those obtained by GC/ECNI‐MS.
Characterization of a novel, testis‐specific equine serine/threonine kinase
Authors: Sabeur Khalida , Ball Barry A. , Corbin C. Jo , Conley Alan
Published: February 2008

Testis‐specific protein kinases are important because of their potential role in spermiogenesis, sperm maturation, and sperm function. In the present study, a novel serine–threonine kinase with high identity to human serine–threonine kinase 31 (STK31) was cloned from equine testis and expression of the protein was characterized in equine testis and ejaculated spermatozoa. Five over‐lapping independent clones were plaque purified after screening of a λ ZAP cDNA expression library constructed from equine testis. Sequence analysis and alignment of all five clones showed high identity with human STK31 with approximately 200 bp of the equine N‐terminal sequence incomplete. The putative full‐length coding sequence of this testis specific equine cDNA was completed by amplification of a 200‐bp fragment using a human primer upstream of the reported translational start site with equine specific nested primers. Northern blot analysis using the equine STK31 cDNA detected an RNA transcript of ∼3.1 kb present in testis but not in other reproductive or somatic tissues. Immunolocalization of the protein in equine testis and spermatozoa demonstrated that STK31 was present in post‐meiotic germ cells with localization to the equatorial segment of testicular spermatozoa. Analysis of the domain structure of equine STK31 revealed a protein kinase domain along with a putative RNA‐binding region. The post‐meiotic expression of this protein along with its domain structure suggests that STK31 may have a role in reorganization of sperm chromatin during spermiogenesis. The cloning of this novel, testis‐specific equine STK provides a new tool to explore the role of kinases in sperm function. Mol. Reprod. Dev. 75: 867–873, 2008. © 2008 Wiley‐Liss, Inc.
Developmental capacity of porcine nuclear transfer embryos correlate with levels of chromatin‐remodeling transcripts in donor cells
Authors: Magnani Luca , Lee Kiho , Fodor William L. , Machaty Zoltan , Cabot Ryan A.
Published: February 2008

Somatic cell nuclear transfer (SCNT) still retains important limitations. Impaired epigenetic reprogramming is considered responsible for altered gene expression and developmental failure in SCNT‐derived embryos. After nuclear transfer the donor cell nucleus undergoes extensive changes in gene expression that involve epigenetic modifications and chromatin remodeling. We hypothesized that SNF2‐type ATP‐dependent chromatin factors contribute to epigenetic reprogramming and the relative amount of these factors in the donor cell affects developmental potential of the reconstructed embryos. In order to test this hypothesis, we assessed the relative amount of SNF2‐type ATPases (Brahma, Brg1, SNF2H, SNF2L, CHD3, and CHD5) in three different donor cells as well as in porcine metaphase II oocytes. We performed SCNT with fetal fibroblast cells, olfactory bulb (OB) progenitor cells, and porcine skin originating sphere stem cells (PSOS). We found that OB‐NT embryos and PSOS‐NT embryos resulted in a higher morulae/blastocysts ratio as compared to fibroblast‐NT embryos (23.53%, 16.98%, and 11.63%, respectively; P < 0.05). Fibroblast cells contained a significantly higher amount of SNF2L and CHD3 transcripts while Brg1 and SNF2H were the most expressed transcripts in all the cell lines analyzed. Metaphase II oocyte expression profile appeared to be unique compared to the cell lines analyzed. This work supports our hypothesis that an array of chromatin‐remodeling proteins on donor cells may influence the chromatin structure, effect epigenetic reprogramming, and developmental potential. Mol. Reprod. Dev. 75: 766–776, 2008. © 2008 Wiley‐Liss, Inc.
A simple mesh deformation technique for fluid–structure interaction based on a submesh approach
Authors: Lefrançois E.
Published: February 2008

This paper describes an improvement in techniques currently used for mesh deformations in fluid–structure calculations in which large body motions are encountered. The proposed approach moving submesh approach (MSA) is based on the assumption of a pseudo‐material deformation applied on a triangular coarse mesh to significantly reduce the CPU time. The computation mesh is then updated using an interpolation technique similar to the finite element method. This method may be applied on structured as well as on unstructured meshes. An extension to complex boundaries undergoing large rigid‐body motions is proposed combining the MSA and an encapsulation box. The influence of the coarse mesh on the quality mesh is discussed. Copyright © 2008 John Wiley & Sons, Ltd.
Comparison of DIGE and post‐stained gel electrophoresis with both traditional and SameSpots analysis for quantitative proteomics
Authors: Karp Natasha A. , Feret Renata , Rubtsov Denis V. , Lilley Kathryn S.
Published: February 2008

2‐DE is an important tool in quantitative proteomics. Here, we compare the deep purple (DP) system with DIGE using both a traditional and the SameSpots approach to gel analysis. Missing values in the traditional approach were found to be a significant issue for both systems. SameSpots attempts to address the missing value problem. SameSpots was found to increase the proportion of low volume data for DP but not for DIGE. For all the analysis methods applied in this study, the assumptions of parametric tests were met. Analysis of the same images gave significantly lower noise with SameSpots (over traditional) for DP, but no difference for DIGE. We propose that SameSpots gave lower noise with DP due to the stabilisation of the spot area by the common spot outline, but this was not seen with DIGE due to the co‐detection process which stabilises the area selected. For studies where measurement of small abundance changes is required, a cost–benefit analysis highlights that DIGE was significantly cheaper regardless of the analysis methods. For studies analysing large changes, DP with SameSpots could be an effective alternative to DIGE but this will be dependent on the biological noise of the system under investigation.
Cartilage proteomics: Challenges, solutions and recent advances
Authors: Wilson Richard , Bateman John F.
Published: February 2008

The mammalian skeleton is largely composed of cartilage and bone. The major functions of cartilage are first to provide a transient template for development of the axial and appendicular skeleton and secondly to provide permanent articulating joint surfaces. The unique cartilage extracellular matrix (ECM) is essential for the load‐bearing and viscoelastic properties of cartilage tissues. Maintained by the chondrocytes, the ECM contains a myriad of proteins and proteoglycans organized into precise networks. Many cartilage disorders result from genetic disruption of cartilage ECM components, their interactions and/or degradation. Although technically challenging, the proteomic analysis of cartilage in development and disease is now emerging as a clinically important research area. In this article, we will review progress in the proteomic characterization of cartilage‐related samples.
Proteomics and naturally occurring animal diseases: Opportunities for animal and human medicine
Authors: Doherty Mary K. , Beynon Robert J. , Whitfield Phillip D.
Published: February 2008

The exquisite sensitivity and selectivity of contemporary protein analysis means that proteomics is increasingly at the forefront of biomedical investigation. The molecular basis of diseases states can now be revealed at the protein level, complementing transcript and genomics data. Moreover, protein biomarkers are increasingly accessible to the scrutiny of the protein chemist. The focus of these studies has been on human systems but diseases that occur naturally in nonhuman mammalian species may provide additional perspectives on the pathophysiology of human disorders. As a result, veterinary research aimed at promoting animal health may lead to concomitant improvements in the characterisation, clinical management and treatment of human diseases. This review will focus on the use of proteomic approaches to study naturally occurring disease states in animals and discuss ways by which these investigations may advance the health of both animals and humans.
Electronic structure and linear optical properties of the double perovskite La2NaRuO6
Authors: Ahmed A. S. , Chen H. , Yuan H. K.
Published: February 2008

First‐principle calculations with the full‐potential linearized augmented plane‐wave (FP‐LAPW) method have been performed to investigate electronic and linear optical properties of the La2NaRuO6 double perovskite. Within the general gradient approximation (GGA), the spin‐polarized calculations predict La2NaRuO6 to be an insulator with antiferromagnetic interactions between the Ru atoms. A gap separation of about 0.64 eV has been shown in the band‐structure feature between the majority and minority spin components. The total spin magnetic moment is about 3.06μB, that mainly comes from Ru atom. The calculated structural parameters and spin moment are consistent with experimental values. The optical constants and energy‐loss spectrum are calculated within the electric‐dipole approximation and the main features shown by the optical function curves can be ascertained on the base of the analysis of the band structure. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Investigation of relaxation stresses in amorphous Pd52Ni32P16
Authors: Melnick A. B. , Djafarov E. J.
Published: February 2008

It is suggested to measure relaxation stresses in amorphous alloys on the basis of diffraction data. Using this procedure, we analysed the elimination of local stresses on low‐temperature annealing of amorphous Pd52Ni32P16. As a result, a three‐dimensional pattern of a stress distribution in raw alloy was obtained, the main feature of which was nanoscale heterogeneities. We found that the value of relaxation energy for Pd52Ni32P16 is 1.49 kJ/mol. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)