bluuebunny/crossref_metadata_2025 huggingface dataset:
Bevacizumab improves pathologic response and protects against hepatic injury in patients treated with oxaliplatin‐based chemotherapy for colorectal liver metastases
Authors: Ribero Dario , Wang Huamin , Donadon Matteo , Zorzi Daria , Thomas Melanie B. , Eng Cathy , Chang David Z. , Curley Steven A. , Abdalla Eddie K. , Ellis Lee M. , Vauthey Jean‐Nicolas
Published: October 2007

BACKGROUND.

The current study evaluated the effect of bevacizumab added to fluoropyrimidine‐plus‐oxaliplatin (5FU/OX) chemotherapy for colorectal liver metastases (CLM) on the pathologic response and nontumorous liver histology.

METHODS.

A total of 105 consecutive patients treated preoperatively with 5FU/OX chemotherapy with (n = 62) or without (n = 43) bevacizumab were analyzed. The response to chemotherapy was evaluated by pathologic analysis of tumor viability (percentage of viable tumor in relation to tumor surface area). The incidence and grade of hepatic sinusoidal dilation were also investigated.

RESULTS.

Bevacizumab‐containing regimens significantly reduced the degree of tumor viability compared with 5FU/OX‐only chemotherapy (32.9% vs 45.3%; P = .02). After stratification according to the magnitude of tumor viability, a higher proportion of patients treated with bevacizumab than without had <25% residual viable tumor cells (45% vs 23%; P = .02). However, the addition of bevacizumab to 5FU/OX did not appear to increase the incidence of complete pathologic response (11.3% vs 11.6%; P = .59). The incidence and severity of sinusoidal dilation was lower in patients treated with bevacizumab than in those treated with 5FU/OX only (any grade: 27.4% vs 53.5%; moderate or severe: 8.1% vs 27.9%; both P < .01).

CONCLUSIONS.

In patients treated with 5FU/OX chemotherapy, bevacizumab improves the pathologic response, as demonstrated by a reduction of the degree of tumor viability, and reduces the incidence and severity of hepatic injury. This retrospective study provides additional evidence supporting the use of bevacizumab in combination with 5FU/OX for CLM. Cancer 2007. © 2007 American Cancer Society.
A comprehensive analytical model of interconnection networks in large‐scale cluster systems
Authors: Javadi Bahman , Abawajy Jemal H. , Akbari Mohammad K.
Published: October 2007

The trends in parallel processing system design and deployment have been toward networked distributed systems such as cluster computing systems. Since the overall performance of such distributed systems often depends on the efficiency of their communication networks, performance analysis of the interconnection networks for such distributed systems is paramount. In this paper, we develop an analytical model, under non‐uniform traffic and in the presence of communication locality, for the m‐port n‐tree family interconnection networks commonly employed in large‐scale cluster computing systems. We use the proposed model to study two widely used interconnection networks flow control mechanism namely the wormhole and store&forward. The proposed analytical model is validated through comprehensive simulation. The results of the simulation demonstrated that the proposed model exhibits a good degree of accuracy for various system organizations and under different working conditions. Copyright © 2007 John Wiley & Sons, Ltd.
High input impedance voltage‐mode first‐order all‐pass sections
Authors: Maheshwari Sudhanshu
Published: October 2007

This paper proposes six new first‐order voltage‐mode all‐pass sections (VM‐APSs) based on three general topologies. Each circuit uses two differential voltage current conveyors and three grounded passive components. All the circuits possess high input impedance and easy control of pole frequency either by a simple matching of resistors (two equal‐valued resistors) for the three canonical circuits or by a single resistor for three non‐canonical circuits. PSPICE simulation results using real device 0.5µ CMOS parameters are given to validate the proposed circuits. Copyright © 2007 John Wiley & Sons, Ltd.
Cationic Gold(I) Phosphanyl Thiolates: Aurophilic Interactions in the Solid State and in Solution
Authors: Balzano Federica , Cuzzola Angela , Diversi Pietro , Ghiotto Fabio , Uccello‐Barretta Gloria
Published: October 2007

The aggregation states in solution of gold(I) cationic complexes of the formula [Au2(StBu)(L2)][BF4] (L = PMe3, PEt3, PtBu3, PPh3; L2 = dppm, dppe) were studied by MS (ESI) and DOSY spectroscopy. These compounds are tetranuclear in the solid state with a core of four gold centres bound together by aurophilic interactions and bridging thiolato groups. Some of these tetranuclear systems are maintained intact in solution depending on the phosphane steric properties or denticity.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
The (Porphyrin)ruthenium‐Catalyzed Aziridination of Olefins Using Aryl Azides as Nitrogen Sources
Authors: Fantauzzi Simone , Gallo Emma , Caselli Alessandro , Piangiolino Cristiana , Ragaini Fabio , Cenini Sergio
Published: October 2007

Aryl azides have been used as atom‐efficient nitrene transfer reagents in the (porphyrin)ruthenium‐catalyzed amination of olefins. Several azides, olefins and [Ru(porphyrin)CO] complexes were tested to investigate the scope and limits of the reaction. Quantitative yields and short reaction times were achieved by using terminal olefins and aryl azides bearing electron‐withdrawing groups on the aryl moiety. The reactions were influenced by steric factors. Internally disubstituted olefins exhibited a lower reactivity and tri‐ and tetrasubstituted olefins did not react at all. A very high turnover number (TON) for the [Ru(TPP)CO] (TPP = tetraphenylporphyrin dianion) catalyzed amination of α‐methylstyrene by p‐nitrophenyl azide was obtained.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
Red Pyrroloquinoline Alkaloids from the Mushroom Mycena haematopus
Authors: Peters Silke , Jaeger Robert J. R. , Spiteller Peter
Published: October 2007

Four so far unknown red alkaloid pigments, haematopodin B (1) and the mycenarubins D (3), E (5) and F (6), were isolated from fruiting bodies of Mycena haematopus. The structures of these pyrroloquinoline alkaloids were established by 2D NMR spectroscopic and MS (ESI) methods. Their absolute configurations were determined by comparison of their CD spectra with that of haematopodin (2) or mycenarubin A (4). Metabolic profiling of the red pigments of intact and injured fruiting bodies revealed that the known degradation product haematopodin (2) originates from haematopodin B (1), which is the native main pigment of M. haematopus.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Rapid analysis of azo‐dyes in food by microchip electrophoresis with electrochemical detection
Authors: Dossi Nicolò , Piccin Evandro , Bontempelli Gino , Carrilho Emanuel , Wang Joseph
Published: October 2007

A method based on microchip electrophoresis with electrochemical (EC) detection has been developed for the simultaneous determination of Yellow AB, Red 2G, Sunset Yellow, New Coccine, and Amaranth which are azo‐dyes frequently added to foodstuffs. Factors affecting both separation and detection processes were examined and optimized, with best performance achieved by using a 10 mM phosphate buffer (pH 11) as BGE solution and applying a voltage of 2500 V both in the separation and in the electrokinetic injection (duration 4 s). Under these optimal conditions, the target dye analytes could be separated and detected within 300 s by applying a detection potential of −1.0 V (vs. Ag/AgCl) to the glassy carbon (GC) working electrode. The recorded peaks were characterized by a good repeatability (RSD = 1.8–3.2%), high sensitivity, and a wide linear range. Detection limits of 3.8, 3.4, 3.6, 9.1, 15.1 μM were obtained for Yellow AB, Red 2G, Sunset Yellow, New Coccine, and Amaranth, respectively. Fast, sensitive, and selective response makes the new microchip protocol very attractive for the quantitative analysis of commercial soft drinks and candies.
Successful transverse patterning in amnesia using semantic knowledge
Authors: Moses Sandra N. , Ostreicher Melanie L. , Rosenbaum R. Shayna , Ryan Jennifer D.
Published: October 2007

We examined whether alternate systems compensate for deficient ones to process relations in amnesia. Transverse patterning (TP), a test of relating items to one another in memory, is reliably impaired in amnesia and has played a central role in testing hippocampal function. We facilitated successful TP in amnesia by providing familiar stimuli with semantically meaningful relationships. Tapping semantic knowledge allowed TP to be solved via extrahippocampal structures. Our work shows that by framing a situation as meaningful we can engage alternate neural systems to compensate for impairments. © 2007 Wiley‐Liss, Inc.
Transgenic inactivation of acetylcholinesterase impairs homeostasis in mouse hippocampal granule cells
Authors: Cohen J.E. , Zimmerman G. , Melamed‐Book N. , Friedman A. , Dori A. , Soreq H.
Published: October 2007

In the adult murine hippocampus, dentate gyrus (DG), neurogenesis and neural cell death are thought to affect learning and memory in incompletely understood mechanism(s). Because cholinergic neurotransmission influences both of these functions, we hypothesized that cholinergic signaling, affected by acetylcholinesterase (AChE) activity, expression level, and alternative splicing, may provide a link between these processes. To challenge this hypothesis, we compared DG neurogenesis in transgenic mice overexpressing engineered “synaptic” AChE‐S, incapable of acetylcholine (ACh) hydrolysis (TgSin) with strain‐matched controls. In control mice, we observed increasing AChE gene expression with progressing neurogenesis. This involved dividing DG neurons expressing proliferating cell nuclear antigen (PCNA) and Tuj1‐positive committed neurons compared with neighboring cells. However, TgSin hippocampi with lower hydrolytic AChE activity showed more PCNA‐labeled cells than controls. In contrast, TgS mice overexpressing catalytically active AChE‐S, with higher than control levels of AChE hydrolytic activity, presented elevated cell labeling by both bromodeoxyuridine and caspase‐3, reflecting facilitated survival of newly born neurons as well as increased neural apoptosis. In comparison, overexpression of the stress‐induced “readthrough” AChE‐R variant in TgR mice resulted in higher hydrolytic activities but unchanged neurogenesis and apoptosis parameters, while all strains presented similar granule cell layer areas, cell density, and neuron numbers. Importantly, this homeostasis was maintained at a cognitive cost: in the hippocampal‐dependent socially transmitted food preference task, TgS and TgSin mice showed impaired acquisition and retention, respectively. Our findings suggest that replacement of AChE‐S with AChE‐R serves to maintain DG homeostasis and associated cognitive tasks, highlighting the role of cholinergic signaling in adult hippocampal neurogenesis and functioning. © 2007 Wiley‐Liss, Inc.
Neurolymphomatosis—diagnosis and assessment of treatment response by FDG PET‐CT
Authors: Lin M , Kilanowska J , Taper J , Chu J
Published: October 2007

Neurolymphomatosis (NL) is lymphomatous infiltration of peripheral nerves, and is an uncommon manifestation of non‐Hodgkin's lymphoma (NHL). Although nerve biopsy is the main method for histological diagnosis, a blind nerve biopsy may not be diagnostic. While CT and MRI have been used to detect NL, recent reports demonstrated the benefit of integrated positron emission tomography (PET) using F18‐2‐fluoro‐2‐deoxy‐D‐glucose (FDG) combined with computed tomography (CT). We described the utility of FDG PET‐CT in this uncommon subgroup of NHL where it can assist in establishing the diagnosis, the potential to guide sites for biopsy and in the assessment of response to therapy. Copyright © 2007 John Wiley & Sons, Ltd.
Self reported sleep quality and cognitive performance in ecstasy users
Authors: Montgomery Catharine , Fisk John E. , Wareing Michelle , Murphy Philip
Published: October 2007

Objectives

Research suggests that ecstasy users exhibit psychobiological changes relative to nonusers such as altered sleep patterns and cognitive deficits. In turn, it has been suggested that sleep quality may be a mediator of such cognitive deficits in ecstasy users. The present study sought to investigate this proposed relationship.

Methods

Aspects of cognitive functioning in 104 ecstasy users and 103 nonusers obtained from our previous studies were reanalysed to explore the extent to which ecstasy‐related group differences were attributable to differences in sleep quality. Cognitive function was assessed via the computation span test, consonant updating, paired associate learning, syllogistic reasoning and word fluency. Sleep quality was measured via the Epworth Sleepiness Scale (ESS), and the Karolinska Sleepiness Scale (KSS).

Results

Ecstasy users performed worse than nonusers on all cognitive measures. While no differences were observed on the ESS, ecstasy users reported greater tiredness at the beginning of testing than nonusers. When the sleep variables were included as covariates, the effects of ecstasy on all cognitive measures remained significant.

Conclusions

The results of the present study suggest little evidence for the mediating effects of sleep on cognitive function in ecstasy users. Copyright © 2007 John Wiley & Sons, Ltd.
Assessing the impact of climate variability and human activities on streamflow from the Wuding River basin in China
Authors: Li Li‐Juan , Zhang Lu , Wang Hao , Wang Juan , Yang Jun‐Wei , Jiang De‐Juan , Li Jiu‐Yi , Qin Da‐Yong
Published: October 2007

Located in the Loess Plateau of China, the Wuding River basin (30 261 km2) contributes significantly to the total sediment yield in the Yellow River. To reduce sediment yield from the catchment, large‐scale soil conservation measures have been implemented in the last four decades. These included building terraces and sediment‐trapping dams and changing land cover by planting trees and improving pastures. It is important to assess the impact of these measures on the hydrology of the catchment and to provide a scientific basis for future soil conservation planning. The non‐parametric Mann–Kendall–Sneyers rank test was employed to detect trends and changes in annual streamflow for the period of 1961 to 1997. Two methods were used to assess the impact of climate variability on mean annual streamflow. The first is based on a framework describing the sensitivity of annual streamflow to precipitation and potential evaporation, and the second relies on relationships between annual streamflow and precipitation. The two methods produced consistent results. A significant downward trend was found for annual streamflow, and an abrupt change occurred in 1972. The reduction in annual streamflow between 1972 and 1997 was 42% compared with the baseline period (1961–1971). Flood‐season streamflow showed an even greater reduction of 49%. The streamflow regime of the catchment showed a relative reduction of 31% for most percentile flows, except for low flows, which showed a 57% reduction. The soil conservation measures reduced streamflow variability, leading to more uniform streamflow. It was estimated that the soil conservation measures account for 87% of the total reduction in mean annual streamflow in the period of 1972 to 1997, and the reduction due to changes in precipitation and potential evaporation was 13%. Copyright © 2007 John Wiley & Sons, Ltd.
Effect of climate changes and water‐related human activities on annual stream flows of the Shiyang river basin in arid north‐west China
Authors: Huo Zailin , Feng Shaoyuan , Kang Shaozhong , Li Wangcheng , Chen Shaojun
Published: October 2007

The change of annual stream flow in the Shiyang river basin, a typical arid‐inland basin in north‐west China, was investigated using hydrological, meteorological and water‐related human activities' data of the past 50 years. The long‐term trends of the hydrological time series were examined by non‐parametric techniques, including the Pettitt and Mann–Kendall tests. Double cumulative curves and multi‐regression methods were used to separate and quantify the effects of climate changes and human activities on the stream flows. The results show that the study area has been experiencing a significant upward warming trend since 1986 and precipitation shows a decreasing trend in the mountainous region but an increasing trend in the plains region. All stream flows in the upper reach and lower reaches of the Shiyang river exhibit decreasing tendencies. Since 1970, human activities, such as irrigation, have had a significant effect on the upstream flow, and account for 60% of total flow decreases in the 1970s. However, climate changes are the main reason for the observed flow decreases in the 1980s and 1990s, with contributions to total flow decrease of 68% and 63%, respectively. Before 1975, flow decreases in the upper reaches were the main factor causing reduced flows in the lower reaches of the Shiyang river. After 1975, the effect of human activities became more pronounced, with contributions of 63%, 68% and 56% to total flow decreases in the lower reaches of the Shiyang river in the periods 1975 to 1980, 1980s and 1990s, respectively. As a result, climate change is responsible for a large proportion of the flow decreases in the upstream section of the catchment during the 1980s and 1990s, while human activities have caused flow decreases downstream during the same period. Copyright © 2007 John Wiley & Sons, Ltd.
Monitoring response to treatment in melanoma patients: Potential of a serum glycomic marker
Authors: Selvan Senthamil R. , Dillman Robert O. , Fowler Abner W. , Carbonell Denysha J. , Ravindranath Mepur H.
Published: October 2007

A mechanistic marker correlating with tumor progression and clinical response is useful for assessing therapeutic response and determining the course of therapy. Since serum‐total‐ganglioside (sTG) and antiganglioside‐IgM antibody levels reflected tumor progression, the feasibility of utilizing sTG for assessing the response to immunotherapy of metastatic‐melanoma was tested. Patients (n = 34) were immunized with dendritic cells cocultured with irradiated, IFNγ‐treated autologous tumor cells admixed with GM‐CSF. Levels of sTG and antiganglioside‐IgM antibody titers were measured in sera of vaccine recipients at 0, 4 and 24 weeks of treatment. Based on sTG‐level, whether lower (L) or higher (H) than the mean + 1 SD of normal and healthy volunteers on weeks 0, 4 and 24, patients were categorized into cohorts‐I (LLL, n = 16), II (HHL/HLL, n = 4), III (LLH/LHH/LHL, n = 7) and IV (HHH/HLH, n = 7). The cohorts were regrouped as sTG‐ downregulators (sTG‐DR; n = 20) and upregulators (sTG‐UR; n = 14). These two cohorts differed significantly in their overall (p < 0.012) and progression‐free (p = 0.0001) survival post‐treatment. 43% sTG‐UR died within 39 months, with a median survival of 39 months, whereas 61% of the sTG‐DR survived for 48 months. Both endogenous and vaccine‐induced antiganglioside‐IgM antibodies appeared to regulate sTG levels. Nonresponders had increased sTG with no or low IgM antibody response. The sTG level is regulated within 24 weeks post‐treatment and therefore, may serve as an ideal biomarker for assessing therapeutic responses in patients. Clinical correlations of sTG indicate that sTG‐downregulating therapy may be an effective treatment strategy for melanoma. © 2007 Wiley‐Liss, Inc.
Ovarian cancers overexpress the antimicrobial protein hCAP‐18 and its derivative LL‐37 increases ovarian cancer cell proliferation and invasion
Authors: Coffelt Seth B. , Waterman Ruth S. , Florez Luisa , Bentrup Kerstin Höner zu , Zwezdaryk Kevin J. , Tomchuck Suzanne L. , LaMarca Heather L. , Danka Elizabeth S. , Morris Cindy A. , Scandurro Aline B.
Published: October 2007

The role of the pro‐inflammatory peptide, LL‐37, and its pro‐form, human cationic antimicrobial protein 18 (hCAP‐18), in cancer development and progression is poorly understood. In damaged and inflamed tissue, LL‐37 functions as a chemoattractant, mitogen and pro‐angiogenic factor suggesting that the peptide may potentiate tumor progression. The aim of this study was to characterize the distribution of hCAP‐18/LL‐37 in normal and cancerous ovarian tissue and to examine the effects of LL‐37 on ovarian cancer cells. Expression of hCAP‐18/LL‐37 was localized to immune and granulosa cells of normal ovarian tissue. By contrast, ovarian tumors displayed significantly higher levels of hCAP‐18/LL‐37 where expression was observed in tumor and stromal cells. Protein expression was statistically compared to the degree of immune cell infiltration and microvessel density in epithelial‐derived ovarian tumors and a significant correlation was observed for both. It was demonstrated that ovarian tumor tissue lysates and ovarian cancer cell lines express hCAP‐18/LL‐37. Treatment of ovarian cancer cell lines with recombinant LL‐37 stimulated proliferation, chemotaxis, invasion and matrix metalloproteinase expression. These data demonstrate for the first time that hCAP‐18/LL‐37 is significantly overexpressed in ovarian tumors and suggest LL‐37 may contribute to ovarian tumorigenesis through direct stimulation of tumor cells, initiation of angiogenesis and recruitment of immune cells. These data provide further evidence of the existing relationship between pro‐inflammatory molecules and ovarian cancer progression. © 2007 Wiley‐Liss, Inc.
Maternal embryonic leucine zipper kinase transcript abundance correlates with malignancy grade in human astrocytomas
Authors: Marie Suely K.N. , Okamoto Oswaldo K. , Uno Miyuki , Hasegawa Ana Paula G. , Oba‐Shinjo Sueli M. , Cohen Tzeela , Camargo Anamaria A. , Kosoy Ana , Carlotti Carlos G. , Toledo Silvia , Moreira‐Filho Carlos A. , Zago Marco A. , Simpson Andrew J. , Caballero Otavia L.
Published: October 2007

We have performed cDNA microarray analyses to identify gene expression differences between highly invasive glioblastoma multiforme (GBM) and typically benign pilocytic astrocytomas (PA). Despite the significant clinical and pathological differences between the 2 tumor types, only 63 genes were found to exhibit 2‐fold or greater overexpression in GBM as compared to PA. Forty percent of these genes are related to the regulation of the cell cycle and mitosis. QT‐PCR validation of 6 overexpressed genes: MELK, AUKB, ASPM, PRC1, IL13RA2 and KIAA0101 confirmed at least a 5‐fold increase in the average expression levels in GBM. Maternal embryonic leucine zipper kinase (MELK) exhibited the most statistically significant difference. A more detailed investigation of MELK expression was undertaken to study its oncogenic relevance. In the examination of more than 100 tumors of the central nervous system, we found progressively higher expression of MELK with astrocytoma grade and a noteworthy uniformity of high level expression in GBM. Similar level of overexpression was also observed in medulloblastoma. We found neither gene promoter hypomethylation nor amplification to be a factor in MELK expression, but were able to demonstrate that MELK knockdown in malignant astrocytoma cell lines caused a reduction in proliferation and anchorage‐independent growth in in vitro assays. Our results indicate that GBM and PA differ by the expression of surprisingly few genes. Among them, MELK correlated with malignancy grade in astrocytomas and represents a therapeutic target for the management of the most frequent brain tumors in adult and children. © 2007 Wiley‐Liss, Inc.
Deregulation of β‐catenin signal by Helicobacter pylori CagA requires the CagA‐multimerization sequence
Authors: Kurashima Yo , Murata‐Kamiya Naoko , Kikuchi Kenji , Higashi Hideaki , Azuma Takeshi , Kondo Satoshi , Hatakeyama Masanori
Published: October 2007

Infection with Helicobacter pylori cagA‐positive strains causes gastritis and peptic ulceration and is associated with gastric adenocarcinoma. The cagA gene product CagA is delivered into gastric epithelial cells, where it undergoes tyrosine phosphorylation by Src family kinases at the C‐terminal EPIYA‐repeat region. Tyrosine‐phosphorylated CagA specifically binds and activates SHP‐2 tyrosine phosphatase, causing cell morphological transformation known as the hummingbird phenotype. CagA also destabilizes the E‐cadherin/β‐catenin complex to elicit aberrant activation of the β‐catenin signal that underlies intestinal metaplasia. Here we show that translocalization of membranous β‐catenin and subsequent activation of the β‐catenin signal by CagA requires the EPIYA‐repeat region, which is characterized by structural variation between CagA of H. pylori isolated in Western countries (Western CagA) and that of East Asian H. pylori isolates (East Asian CagA), but is independent of CagA tyrosine phosphorylation. Detailed analysis using a series of Western and East Asian CagA mutants revealed that deregulation of β‐catenin requires residues 1009–1086 and residues 908–1012 of ABCCC Western CagA and ABD East Asian CagA, respectively, and is mediated by the 16‐amino‐acid CagA multimerization sequence that is conserved between the 2 geographically distinct H. pylori CagA species. Our results indicate that aberrant activation of the β‐catenin signal, which promotes precancerous intestinal metaplasia, is an inherent and fundamental CagA activity that is independent of the structural polymorphism of CagA. © 2007 Wiley‐Liss, Inc.
Osteopontin‐c is a selective marker of breast cancer
Authors: Mirza Mana , Shaughnessy Elizabeth , Hurley John K. , Vanpatten Kristie A. , Pestano Gary A. , He Bin , Weber Georg F.
Published: October 2007

While the acquisition of invasiveness is a critical step in early stage breast carcinomas (DCIS), no established molecular markers reliably identify tumor progression. The metastasis gene osteopontin is subject to alternative splicing, which yields 3 messages, osteopontin‐a, osteopontin‐b and osteopontin‐c. Osteopontin‐c is selectively expressed in invasive, but not in noninvasive, breast tumor cell lines, and it effectively supports anchorage independence. We evaluated osteopontin‐c as a biomarker. The RNA message for osteopontin‐c was present in 16 of 20 breast cancers (80%), but was undetectable in 22 normal specimens obtained from reduction mammoplasty. In contrast, osteopontin‐a RNA was expressed at various levels in all 20 breast cancers, 11 tumor‐surrounding tissues and 21 normal samples. The splice variant osteopontin‐b was present at barely detectable levels in 18 of 20 cancers and in 6 of 22 normal breasts. By immunohistochemistry, 66 of 69 normal breasts were negative, while 3 showed low level staining. Among the breast cancers, 43 of 56 cores (77%) stained positive for osteopontin‐c. When correlated with tumor grade, the staining for osteopontin‐c increased from grade 1 to grade 3. In a total of 178 breast specimens analyzed, osteopontin‐c was present in 78% of cancers, 36% of surrounding tissues and 0% of normal tissues. Furthermore, osteopontin‐c detects a higher fraction of breast cancers than estrogen receptor (ER), progesterone receptor or HER2. In conjunction, osteopontin‐c, ER and HER2 reliably predict grade 2–3 breast cancer. Hence, osteopontin‐c is a diagnostic and prognostic marker that may have value in a diagnostic panel together with conventional breast cancer markers. © 2007 Wiley‐Liss, Inc.
BCL‐2 functions as an activator of the AKT signaling pathway in pancreatic cancer
Authors: Mortenson Melinda M. , Galante Joseph G. , Gilad Oren , Schlieman Michael G. , Virudachalam Subbulakshmi , Kung Hsing‐Jien , Bold Richard J.
Published: October 2007

BCL‐2 is the prototypic anti‐apoptotic protein involved in the regulation of apoptosis. Overexpression of BCL‐2 is common in pancreatic cancer and confers resistance to the apoptotic effect of chemo‐ and radiotherapy. Although these cellular effects of BCL‐2 are traditionally related to pathways involving the mitochondrial membrane, we sought to investigate whether BCL‐2 is involved in other signaling pathways regulating cell survival and focused on AKT. We examined the effect of overexpression of BCL‐2 in the MIA‐PaCa‐2 human pancreatic cancer cell line on the function and subcellular location of AKT. We observed that the stable subclones of MIA‐PaCa‐2 overexpressing BCL‐2 demonstrated increased activity of AKT as well as IKK (a downstream target of AKT), increasing the transcriptional activity of NF‐κB. Using immunoprecipitation techniques, we observed co‐immunoprecipitation of AKT and BCL‐2. Immunocytochemistry demonstrated co‐localization of BCL‐2 and AKT, which was abrogated by treatment with HA14‐1, a small molecule inhibitor of BH‐3‐mediated protein interaction by BCL‐2. Furthermore, treatment with HA14‐1 decreased phosphorylation of AKT and increased sensitivity to the apoptotic effect of the chemotherapeutic agent, paclitaxel. These results demonstrate an additional mechanism of regulation of cell survival mediated by BCL‐2, namely through AKT activation, in the MIA‐PaCa‐2 pancreatic cancer cell line. Therefore, directed inhibition of BCL‐2 may alter diverse pathways controlling cell survival and overcome the apoptotic resistance that is the hallmark of pancreatic cancer. J. Cell. Biochem. 102: 1171–1179, 2007. © 2007 Wiley‐Liss, Inc.
Differential expression of estrogen receptor alpha (ERα) protein in MCF‐7 breast cancer cells chronically exposed to TCDD
Authors: Marquez‐Bravo Lydia G. , Gierthy John F.
Published: October 2007

Estrogens play a key role in the development and evolution of breast cancer tumors. Estrogen receptor alpha (ERα) mediates many of the biological activities of estrogens, and its expression is associated with low invasiveness and good prognosis. Recent epidemiological reports suggest that long‐term exposure to 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) is implicated in the increased incidence of breast cancer in exposed women. TCDD interferes with the expression of some ERα‐dependent genes and inhibits estradiol (E2)‐ dependent growth of breast cancer cells in vitro. However, E2‐dependent xenographs of MCF‐7 human breast cancer cells resumed growth after a 2‐week exposure to TCDD. The mechanisms involved in the resumption of cell growth are not completely understood. In this study, we show that short term‐exposure (16 days) to 1 nM TCDD results in the suppression of ERα protein expression, while chronic exposure for more than 1 year (LTDX cells) results in the partial re‐expression of the receptor. Immunocytochemistry studies showed that re‐expression of ERα in LTDX cells occurred in some of the cells. Analysis by Western immunoblots indicated that four out of five LTDX clones expressed ERα at levels comparable to those in unexposed MCF‐7 cells. Removal of TCDD treatment for 16 days restored the expression of ERα in the ERα‐negative clonal cells. These results suggest that MCF‐7 cells chronically exposed to TCDD contain at least two cell subpopulations that may respond differently to the ERα‐mediated effects of TCDD. J. Cell. Biochem. 103: 636–647, 2008. © 2007 Wiley‐Liss, Inc.
Atrial natriuretic peptide attenuates high glucose‐activated transforming growth factor‐β, Smad and collagen synthesis in renal proximal tubular cells
Authors: Lo Chao‐Sheng , Chen Zhao‐Hong , Hsieh Tusty‐Jiuan , Shin Shyi‐Jang
Published: October 2007

Atrial natriuretic peptide, besides its role in the regulation of volume homeostasis, has been noted to exert cytoprotective effects in several cell types from hypoxia. The present study was performed to explore the effect of ANP on high glucose‐activated transforming growth factor‐beta1 (TGF‐β1), Smad and collagen synthesis in renal proximal epithelial cells. Cultured NRK‐52E cells were divided into five groups: (1) normal glucose (5.5 mM), (2) high glucose (35 mM), (3) D‐mannitol (29.5 mM), (4) high glucose plus ANP (10−6–10−9 M), and (5) high glucose plus ANP (10−6 M) and guanylate cyclase inhibitor LY83583 (10−7 M) groups. Messenger RNA levels of TGF‐β1, Smad2, and collagens were measured by RT‐PCR. ELISA, immunocytochemistry and Western blotting were used to detect protein levels of TGF‐β1, Smad2, phospho‐Smad 2/3 and collagen type 1. We found high glucose to significantly increase mRNA levels of TGF‐β1, Smad 2, collagen types I and III and protein levels of TGF‐β1, phospho‐Smad 2/3 and collagen type 1, but mannitol did not affect their expression. The addition of ANP significantly attenuated high glucose‐enhanced mRNA and protein levels of TGF‐β1, Smad and collagens. LY83583 blocked the influence of ANP on high glucose‐activated TGF‐β1, Smad and collagen synthesis. This is the first study to demonstrate that activation of TGF‐β1, Smad and collagen synthesis stimulated by high glucose can also be inhibited by exogenous ANP in renal tubular epithelial cells. J. Cell. Biochem. 103: 1999–2009, 2008. © 2007 Wiley‐Liss, Inc.
Immortalized mouse articular cartilage cell lines retain chondrocyte phenotype and respond to both anabolic factor BMP‐2 and pro‐inflammatory factor IL‐1
Authors: Majumdar Manas K. , Chockalingam Priya S. , Bhat Ramesh A. , Sheldon Richard , Keohan Cristin , Blanchet Tracey , Glasson Sonya , Morris Elisabeth A.
Published: October 2007

Articular cartilage chondrocytes help in the maintenance of tissue homeostasis and function of the articular joint. Study of primary chondrocytes in culture provides information closely related to in vivo functions of these cells. Limitations in the primary culture of chondrocytes have lead to the development of cells lines that serve as good surrogate models for the study of chondrocyte biology. In this study, we report the establishment and characterization of chondrocyte cell lines, MM‐Sv/HP and MM‐Sv/HP‐2 from mouse articular cartilage. Cells were isolated from mouse femoral head articular cartilage, immortalized and maintained in culture through numerous passages. The morphology of the cells was from fibroblastic to polygonal in nature. Gene expression studies using quantitative PCR (Q‐PCR) were performed on cells in monolayer culture and cells embedded in a three‐dimensional alginate matrix. Stimulation of cells in monolayer culture with anabolic factor, BMP‐2, resulted in increased gene expression of the extracellular matrix molecules, aggrecan and type II collagen and their regulator transcription factor, Sox9. Treatment by pro‐inflammatory IL‐1 resulted in increased gene expression of catabolic effectors including Aggrecanases (ADAMTS4, ADAMTS5), MMP‐13 and nitric oxide synthase (Nos2). Cells in alginate treated with BMP‐2 resulted in increased synthesis of proteoglycan which was released into the conditioned media on IL‐1 stimulation. Western analysis of conditioned media showed the presence of Aggrecanase‐cleaved aggrecan fragments. In summary, MM‐Sv/HP and MM‐Sv/HP‐2 show preservation of important characteristics of articular chondrocytes as examined under multiple culture conditions and would provide a useful reagent in the study of chondrocyte biology. J. Cell. Physiol. 215: 68–76, 2008. © 2007 Wiley‐Liss, Inc.
Pleiotrophin induces nitric oxide dependent migration of endothelial progenitor cells
Authors: Heiss Christian , Wong Maelene L. , Block Vanessa I. , Lao David , Real Wendy May , Yeghiazarians Yerem , Lee Randall J. , Springer Matthew L.
Published: October 2007

Pleiotrophin (PTN) is produced under ischemic conditions and has been shown to induce angiogenesis in vivo. We studied whether or not PTN exerts chemotaxis of pro‐angiogenic early endothelial progenitor cells (EPCs), a population of circulating cells that have been reported to participate in and stimulate angiogenesis. Chemotaxis of EPCs, isolated from blood of healthy humans (n = 5), was measured in transwell assays. PTN at 10–500 ng/ml elicited dose‐dependent chemotaxis of both EPCs and human umbilical vein endothelial cells (HUVECs), but not of human coronary artery smooth muscle cells (CASMCs) and T98G glioblastoma cells that lack PTN receptors. The degree of chemotaxis was comparable to that induced by the angiogenic factors VEGF and SDF‐1α. Chemotaxis to PTN was blocked by the NOS inhibitors L‐NNA and L‐NMMA, the NO scavenger PTIO, the phosphoinositide‐3 kinase inhibitor wortmannin, and the guanylyl cyclase inhibitor ODQ, suggesting dependence of EPC chemotaxis on these pathways. PTN induced NOS‐dependent production of NO to a similar degree as did VEGF, as indicated by the NO indicator DAF‐2. PTN increased proliferation in EPCs and HUVECs to a similar extent as VEGF, but did not induce proliferation of CASMCs. While L‐NNA abolished PTN‐induced migration in EPCs and HUVECs, it did not inhibit PTN‐ and VEGF‐enhanced proliferation and also caused proliferation by itself. These data suggest that PTN may mediate its pro‐angiogenic effects by increasing the local number of not only endothelial cells but also early EPCs at angiogenic sites. J. Cell. Physiol. 215: 366–373, 2008. © 2007 Wiley‐Liss, Inc.
RETRACTED: Basic fibroblast growth factor accelerates matrix degradation via a neuro‐endocrine pathway in human adult articular chondrocytes
Authors: Im Hee‐Jeong , Li Xin , Muddasani Prasuna , Kim Gun‐Hee , Davis Francesca , Rangan Jayanthi , Forsyth Christopher B. , Ellman Michael , Thonar Eugene JMA
Published: October 2007

Pain‐related neuropeptides released from synovial fibroblasts, such as substance P, have been implicated in joint destruction. Substance P‐induced inflammatory processes are mediated via signaling through a G‐protein‐coupled receptor, that is, neurokinin‐1 tachykinin receptor (NK1‐R). We determined the pathophysiological link between substance P and its receptor in human adult articular cartilage homeostasis. We further examined if catabolic growth factors such as basic fibroblast growth factor (bFGF or FGF‐2) or IL‐1β accelerate matrix degradation via a neural pathway upregulation of substance P and NK1‐R. We show here that substance P stimulates the production of cartilage‐degrading enzymes, such as matrix metalloproteinase‐13 (MMP‐13), and suppresses proteoglycan deposition in human adult articular chondrocytes via NK1‐R. Furthermore, we have demonstrated that substance P negates proteoglycan stimulation promoted by bone morphogenetic protein‐7, suggesting the dual role of substance P as both a pro‐catabolic and anti‐anabolic mediator of cartilage homeostasis. We report that bFGF‐mediated stimulation of substance P and its receptor NK1‐R is, in part, through an IL‐1β‐dependent pathway. J. Cell. Physiol. 215: 452–463, 2008. © 2007 Wiley‐Liss, Inc.
Melanocytes expressing MC1R polymorphisms associated with red hair color have altered MSH‐ligand activated pigmentary responses in coculture with keratinocytes
Authors: Roberts Donald W. , Newton Richard A. , Leonard J. Helen , Sturm Richard A.
Published: October 2007

The occurrence of red hair and pale skin in individuals, which is associated with UV‐radiation sensitivity and increased skin cancer risk, is mainly due to polymorphisms in the melanocortin‐1 receptor (MC1R) expressed in melanocytes. We have established a serum free human melanocyte–keratinocyte coculture system to study the behavior and functional abilities of melanocytes expressing MC1R red hair color (RHC) variants in order to identify differences from their wild type (WT) counterparts. This model revealed the importance of elevated calcium levels in promoting strong melanocyte interaction with the surrounding keratinocytes and resulted in a dendritic melanocyte morphology similar to that in skin. However, the dendricity response following agonist activation of the MC1R receptor by NDP‐MSH peptide, was markedly enhanced in WT melanocytes in comparison to RHC strains. Analysis of mRNA expression and protein levels of the major pigmentation markers following NDP‐MSH treatment distinguished the enzyme dopachrome tautomerase as preferentially upregulated in cocultures of WT strains, with negligible or a much reduced response in melanocytes with RHC variant alleles. These results highlight the use of the coculture system in determining fundamental differences in the physiology of melanocytes expressing RHC MC1R receptors and those of WT genotype, which are likely to contribute to the increased skin cancer risk for individuals that carry these variants. J. Cell. Physiol. 215: 344–355, 2008. © 2007 Wiley‐Liss, Inc.
Progression elevated gene‐3 promoter (PEG‐Prom) confers cancer cell selectivity to human polynucleotide phosphorylase (hPNPaseold‐35)‐mediated growth suppression
Authors: Chan Isaac , Lebedeva Irina V. , Su Zao‐Zhong , Sarkar Devanand , Valerie Kristoffer , Fisher Paul B.
Published: October 2007

The poor prognosis of pancreatic cancer patients using currently available therapies mandates novel therapeutics that combine anti‐neoplastic potency with toxicity‐minimizing cancer specificity. Employing an overlapping pathway screen to identify genes exhibiting coordinated expression as a consequence of terminal cell differentiation and replicative senescence, we identified human polynucleotide phosphorylase (hPNPaseold‐35), a 3′,5′‐exoribonuclease that exhibits robust growth‐suppressing effects in a wide spectrum of human cancers. A limitation to the anti‐neoplastic efficacy of hPNPaseold‐35 relates to its lack of cancer specificity. The promoter of Progression Elevated Gene‐3 (PEG‐Prom), discovered in our laboratory via subtraction hybridization in a transformation progression rodent tumor model functions selectively in a diverse array of human cancer cells, with limited activity in normal cells. An adenovirus constructed with the PEG‐Prom driving expression of hPNPaseold‐35 containing a C‐terminal Hemaglutinin (HA)‐tag (Ad.PEG.hPNPaseold‐35) was shown to induce robust transgene expression, growth suppression, apoptosis, and cell‐cycle arrest in a broad panel of pancreatic cancer cells, with minimal effects in normal immortalized pancreatic cells. hPNPaseold‐35 expression correlated with arrest in the G2/M phase of the cell cycle and up‐regulation of the cyclin‐dependent kinase inhibitors (CDKI) p21CIP1/WAF‐1/MDA‐6 and p27KIP1. In a nude mouse xenograft model, Ad.PEG.hPNPaseold‐35 injections effectively inhibited growth of human pancreatic cancer cells in vivo. These findings support the potential efficacy of combining a cancer‐specific promoter, such as the PEG‐Prom, with a novel anti‐neoplastic agent, such as hPNPaseold‐35, to create a potent, targeted cancer therapeutic, especially for a devastating disease like pancreatic cancer. J. Cell. Physiol. 215: 401–409, 2008. © 2007 Wiley‐Liss, Inc.
Osteoblast proliferation or differentiation is regulated by relative strengths of opposing signaling pathways
Authors: Raucci Angela , Bellosta Paola , Grassi Roberta , Basilico Claudio , Mansukhani Alka
Published: October 2007

Skeletal development requires the correct balance of osteoblast proliferation, survival, and differentiation which is modulated by a network of signaling pathways and transcription factors. We have examined the role of the AKT (PKB), and ERK1/2 signaling pathways in the osteoblast response to FGFs, which inhibit differentiation, and to IGF‐1 and Wnt signaling, which promote it. Using osteoblastic cell lines as well as primary calvarial osteoblasts, we show that ERK1/2 and AKT have distinct effects in FGF‐induced osteoblast proliferation and differentiation. ERK1/2 is a primary mediator of FGF‐induced proliferation, but also contributes to osteoblast differentiation, while AKT is important for osteoblast survival. Signaling by IGF‐1, that promotes osteoblast differentiation, strongly activates AKT and weakly ERK1/2, while the opposite results are obtained with FGF, which inhibits differentiation. By introducing a constitutively active form of AKT, we found that increased AKT activity drives osteoblasts to differentiation. Increasing the AKT signal in osteoblasts that harbor FGFR2 activating mutations, found in Crouzon (342Y) and Apert (S22W) syndromes, is also able to drive differentiation in these cells, that normally fail to differentiate. Wnt signals, that promotes differentiation, also induce AKT phosphorylation, and cells expressing active AKT have increased levels of stabilized beta‐catenin, a central molecule in Wnt signaling. Our results indicate that the relative strengths of ERK and AKT signaling pathways determine whether osteoblasts are driven into proliferation or differentiation, and that the effects of AKT may be due, in part, to synergy with the Wnt pathway as well as with the Runx2 transcription factor. J. Cell. Physiol. 215: 442–451, 2008. © 2007 Wiley‐Liss, Inc.
Identification of the gene transcription and apoptosis mediated by TGF‐β‐Smad2/3‐Smad4 signaling
Authors: Yu Jian , Zhang Liang , Chen Ailiang , Xiang Guangxin , Wang Yahui , Wu Jianping , Mitchelson Keith , Cheng Jing , Zhou Yuxiang
Published: October 2007

Transforming growth factor‐β (TGF‐β) signaling is known to depend on the formation of Smad2/3‐Smad4 transcription regulatory complexes. However, the signaling functions of Smad2/3‐Smad4 during TGF‐β‐induced responses are obscure as TGF‐β also initiates a number of other signaling pathways. In this study, we systematically assessed the contribution of TGF‐β‐Smad2/3‐Smad4 signaling to both target gene transcription and apoptosis. Individual Smads were selectively knocked down in Hep3B cells by stable RNA interference (RNAi). We identified TGF‐β‐responsive genes using genome‐wide oligonucleotide microarrays and confirmed their dependency on Smad2, Smad3, or Smad4 by the combination of RNAi and microarray assay. The major finding from our microarray analysis was that of the 2,039 target genes seen to be regulated via TGF‐β induction, 190 were differentially transcriptionally controlled by Smad2‐Smad4 and Smad3‐Smad4 signaling and the latter control mechanism appeared to be functionally more important. We also found indirect evidence of competition between Smad2 and Smad3 for their activation when controlling the transcription of target genes. Functional analysis revealed that Smad3 and Smad4 were the predominant mediators of TGF‐β‐induced apoptosis in Hep3B cells. We provide evidence that up‐regulation of Bcl‐2‐interacting mediator of cell death (Bim), under the transcriptional control of Smad3‐Smad4 signaling, is crucial to TGF‐β‐induced apoptosis in Hep3B cells. J. Cell. Physiol. 215: 422–433, 2008. © 2007 Wiley‐Liss, Inc.
Proteomic analysis of metacyclic trypomastigotes undergoing Trypanosoma cruzi metacyclogenesis
Authors: Parodi‐Talice A. , Monteiro‐Goes V. , Arrambide N. , Avila A. R. , Duran R. , Correa A. , Dallagiovanna B. , Cayota A. , Krieger M. , Goldenberg S. , Robello C.
Published: October 2007

Trypanosoma cruzi, the causative agent of the Chagas disease, has a complex life cycle alternating between replicative and noninfective forms with nonreplicative and infective forms of the parasite. Metacyclogenesis is a process that takes place in the invertebrate host, comprising morphogenetic transformation from a noninfective form to an infective form, such that parasites acquire the ability to invade human cells. We analyze here the metacyclogenesis process by 2D electrophoresis coupled to MALDI‐TOF MS. A large proportion of unique proteins expressed during metacyclogenesis were observed. Interestingly, 50% of the spots were found to differ between epimastigotes and trypomastigotes. We provide a 2D map of the infective metacyclic trypomastigotes. Sixty six protein spots were successfully identified corresponding to 43 different proteins. We analyzed the expression profiles for the identified proteins along metacyclogenesis and classified them into three groups according to their maximal level of expression. We detected several isoforms for a number of proteins, some displaying differential expression during metacyclogenesis. These results suggest that posttranslational modifications may be a fundamental part of the parasite's strategy for regulating gene expression during differentiation. This study contributes to the identification of relevant proteins involved in the metacyclogenesis process. The identification and molecular characterization of these proteins will render vital information about the steps of the parasite differentiation into the infective form. Copyright © 2007 John Wiley & Sons, Ltd.