bluuebunny/crossref_metadata_2025 huggingface dataset:
The cisplatin, epirubicin, 5‐fluorouracil, gemcitabine (PEFG) regimen in advanced biliary tract adenocarcinoma
Authors: Cereda Stefano , Passoni Paolo , Reni Michele , Viganò Maria G. , Aldrighetti Luca , Nicoletti Roberto , Villa Eugenio
Published: February 2010

BACKGROUND:

Biliary tract adenocarcinoma (BTA) is an uncommon tumor with a poor prognosis and no standard, systemic chemotherapy. The combined cisplatin, epirubicin, 5‐fluorouracil, and gemcitabine (PEFG) regimen is an effective, upfront treatment for advanced pancreatic cancer. In this study, the authors assessed the activity and safety of this combination regimen in patients with advanced BTA.

METHODS:

PEFG (cisplatin 40 mg/m2 and epirubicin 40 mg/m2 on Day 1; gemcitabine 600 mg/m2 on Days 1 and 8; and 5‐fluorouracil [FU] 200 mg/m2 daily as a continuous infusion) was administered to chemotherapy‐naive patients who had a cytologic or histologic diagnosis of locally advanced or metastatic BTA, aged ≤75 years, and a performance status (PS) >60 either until they had evidence progressive disease or for a maximum of 6 months. Tumor size was assessed every 2 months during treatment.

RESULTS:

Between May 1999 and December 2005, 37 patients (62% metastatic) who had a median age of 62 years and a median PS of 90 received the PEFG regimen at the authors' institution. Primary tumor sites were the intrahepatic bile duct in 10 patients (27%), the extrahepatic bile duct in 8 patients (22%), the gallbladder in 12 patients (32%), and the ampulla of Vater in 7 patients (19%). A partial response was observed in 16 patients (43%), and stable disease was observed in 12 patients (32%). The median overall survival (OS) was 12.1 months, and the 1‐year OS rate was 52%. The median progression‐free survival (PFS) was 7.9 months, and the 6‐month PFS rate was 67%. The main grade 3/4 toxicity was neutropenia in 18% of cycles followed by thrombocytopenia in 9% of cycles, nausea/vomiting in 5% of cycles, and febrile neutropenia, fatigue, anemia, and stomatitis in 2% of cycles.

CONCLUSIONS:

The current results demonstrated that PEFG was an active regimen with a manageable toxicity profile for patients with advanced BTA. Cancer 2010. © 2010 American Cancer Society.
A new era in anticancer peptide vaccines
Authors: Perez Sonia A. , von Hofe Eric , Kallinteris Nikoletta L. , Gritzapis Angelos D. , Peoples George E. , Papamichail Michael , Baxevanis Constantin N.
Published: February 2010

The use of synthetic peptides as vaccines aimed at the induction of therapeutic CD8‐positive T‐cell responses against tumor cells initially experienced great enthusiasm, mostly because of advances in vaccine technology, including design, synthesis, and delivery. However, despite impressive results in animal models, the application of such vaccines in humans has met with only limited success. The therapeutic activity of vaccine‐stimulated, tumor‐specific, CD8‐positive T cells can be hampered through the physical burden of the tumor, tolerance mechanisms, and local factors within the tumor microenvironment. Recently, accumulating evidence has suggested that combining a peptide‐based therapeutic vaccination with conventional chemotherapy can uncover the full potential of the antitumor immune response, increasing the success of immunotherapy. In addition, therapeutic vaccination in the preventive setting has been extremely effective in eliciting antitumor responses in preclinical tumor models and has demonstrated good promise clinically in patients with minimal residual disease. The rationale behind preventive vaccination is that patients with minimal tumor burden still have a fully competent immune system capable of developing robust antitumor responses. Finally, therapeutic CD8‐positive T‐cell peptide vaccines have been improved by coimmunization with T‐helper epitopes expressed on long peptides. Cancer 2010. © 2010 American Cancer Society.
Predictors of weight loss during radiotherapy in patients with stage I or II head and neck cancer
Authors: Nourissat Alice , Bairati Isabelle , Samson Elodie , Fortin André , Gélinas Michel , Nabid Abdenour , Brochet François , Têtu Bernard , Meyer François
Published: February 2010

BACKGROUND:

The purpose of the study was to identify predictors of weight loss during radiotherapy (RT) in patients with stage I or II head and neck (HN) cancer.

METHODS:

This study was conducted as part of a phase 3 chemoprevention trial. A total of 540 patients were randomized. The patients were weighed before and after RT. Their baseline characteristics, including lifestyle habits, diet, and quality of life, were assessed as potential predictors. Predictors were identified using multiple linear regressions. The reliability of the model was assessed by bootstrap resampling. A receiver operating characteristics curve was generated to estimate the model's accuracy in predicting critical weight loss (≥5%).

RESULTS:

The mean weight loss was 2.2 kg (standard deviation, 3.4). Five factors were associated with a greater weight loss: all HN cancer sites other than the glottic larynx (P<.001), higher pre‐RT body weight (P<.001), stage II disease (P = .002), dysphagia and/or odynophagia before RT (P = .001), and a lower Karnofsky performance score (P = .028). There was no association with pre‐RT lifestyle habits, diet, or quality of life. The bootstrapping method confirmed the reliability of this predictive model. The area under the curve was 71.3% (95% confidence interval, 65.8‐76.9), which represents an acceptable ability of the model to predict critical weight loss.

CONCLUSIONS:

These results could be useful to clinicians for screening patients with early stage HN cancer treated by RT who require special nutritional attention. Cancer 2010. © 2010 American Cancer Society.
Preparation of 9α‐Fluorinated Sesquiterpenic Drimanes and Evaluation of Their Antifeedant Activities
Authors: Abad Antonio , Agulló Consuelo , Cuñat Ana C. , González‐Coloma Azucena , Pardo David
Published: February 2010

The preparation of 9α‐fluoro analogues of both natural and unnatural drimane‐type sesquiterpenes is described. Their synthesis began with the initial preparation of methyl 8‐keto‐12‐nordriman‐11‐oate from β‐ionone and entailed the electrophilic fluorination of C‐9 for the stereoselective introduction of the fluorine atom. The drimane skeleton was completed from the intermediate 9α‐fluoro‐8‐keto‐12‐nordrimane system by means of different reactions at the C‐8 ketone carbonyl group, essentially Wittig methylenation, cyanohydrin formation or palladium‐catalysed carbonylation of the corresponding enol triflate. Further manipulation of the functionalization derived from these key reactions allowed the preparation, among others, of 9α‐fluorodrimanes, which are structurally and functionally related to albicanic acid, drimenin and olepupuane. Also described are the reactivities of some of the fluorine‐containing systems prepared and a comparative study of the antifeedant activities of a selection of 9α‐fluorodrimanes and the corresponding hydrogen analogues against several insect species with different feeding ecologies (Spodoptera littoralis,Myzus persicaeandRhopalosiphum padi), which revealed a significant increase in the antifeedant activities of some of the fluorinated drimane analogues.
Environmental occurrence and reproductive effects of the pharmaceutical fluoxetine in native freshwater mussels
Authors: Bringolf Robert B. , Heltsley Rebecca M. , Newton Teresa J. , Eads Chris B. , Fraley Stephen J. , Shea Damian , Cope W. Gregory
Published: February 2010

The present study measured the occurrence, distribution, and bioaccumulation of fluoxetine in samples of water, polar organic chemical integrative sampler (POCIS), sediment, and caged freshwater mussels at stream sites near a municipal wastewater treatment facility effluent discharge. We assessed the relation of the environmental concentrations to reproductive endpoints in mussels in acute laboratory tests. Concentrations of fluoxetine in water and POCIS samples were similar (<20% difference) within each site and were greatest in the effluent channel (104–119 ng/L), and decreased at 50 m and 100 m downstream. Likewise, concentrations of fluoxetine in sediment and mussel (Elliptio complanata) tissue were greatest in the effluent channel (17.4 ng/g wet wt for sediment and 79.1 ng/g wet wt for mussels). In 96-h lab tests, fluoxetine significantly induced parturition of nonviable larvae from female E. complanata exposed to 300 µg/L (p = 0.0118) and 3,000 µg/L (p < 0.0001) compared to controls. Fluoxetine exposure at 300 µg/L (p = 0.0075) and 3,000 µg/L (p = 0.0001) also resulted in stimulation of lure display behavior in female Lampsilis fasciola and Lampsilis cardium, respectively. In male E. complanata, 3,000 µg fluoxetine/L significantly induced release of spermatozeugmata during a 48-h exposure. These results suggest that fluoxetine accumulates in mussel tissue and has the potential to disrupt several aspects of reproduction in freshwater mussels, a faunal group recognized as one of the most imperiled in the world. Despite the disparity between measured environmental concentrations of fluoxetine and effects concentrations in our short-term tests with these long-lived animals, additional tests are warranted to evaluate the effects of long-term exposure to environmentally relevant concentrations and critical lifestages (e.g., juveniles). Environ. Toxicol. Chem. 2010;29:1311–1318. © 2010 SETAC
Excisional biopsy with margin control for oral cancers
Authors: Wang Yu‐Chien , Fang Ku‐Hao , Jung Shih‐Ming , Zheng Jie , Hao Sheng‐Po
Published: February 2010

Background

This study was undertaken to show how an excisional biopsy procedure can be used to remove the entire specimen for oral lesions with difficult diagnoses.

Methods

We retrospectively reviewed 52 cases with 56 oral lesions either suspected to be malignant or with a preliminary controversial diagnosis. These were treated by excisional biopsy with frozen section intraoperatively between January 2003 and June 2008.

Results

Twenty‐eight of 56 lesions were diagnosed with malignancy. We found a high underdiagnosis rate (7 of 12) in patients who had a preliminary controversial diagnosis. Twenty‐one patients without preoperative biopsy were diagnosed to have malignancies. Of the 28 malignant cases, all surgical margins were negative. Neck ultrasound scans with fine‐needle aspiration cytology was carried out and no patient demonstrated simultaneous neck metastasis. One patient died of locoregional recurrence. Two patients died of a second primary cancer.

Conclusion

Excisional biopsy with margin control is useful both in diagnosis and treatment of oral cancers. © 2010 Wiley Periodicals, Inc. Head Neck, 2010
Chemoradiotherapy for locoregionally advanced squamous cell carcinoma of the base of tongue
Authors: Pederson Aaron W. , Haraf Daniel J. , Witt Mary‐Ellyn , Stenson Kerstin M. , Vokes Everett E. , Blair Elizabeth A. , Salama Joseph K.
Published: February 2010

Background

Our aim was to report the outcomes of base of tongue cancers treated with chemoradiotherapy.

Methods

Between 1990 and 2004, 127 patients with stage III or IV base of tongue cancer were treated with chemoradiotherapy on protocol. Indications included nodal involvement, T3/T4 tumors, positive margins, those patients refusing surgery, or were medically inoperable. The most common regimen was paclitaxel (100 mg/m2 on day 1), infusional 5‐fluorouracil (600 mg/m2/day × 5 days), hydroxyurea (500 mg prescribed orally [PO] 2 × daily [BID]), and 1.5 Gy twice daily irradiation followed by a 9‐day break without treatment.

Results

Median follow‐up was 51 months. The median dose to gross tumor was 72.5 Gy (range, 40–75.5 Gy). Five‐year locoregional progression‐free survival, overall survival, and disease‐free survival was 87.0%, 58.2%, and 46.0%, respectively.

Conclusion

Concurrent chemoradiotherapy results in promising locoregional control for base of tongue cancer. As distant relapse was common, further investigation of systemic therapy with novel agents may be warranted. © 2010 Wiley Periodicals, Inc. Head Neck, 2010
The effects of telomerase inhibition on prostate tumor‐initiating cells
Authors: Marian Calin O. , Wright Woodring E. , Shay Jerry W.
Published: February 2010

Prostate cancer is the most common malignancy in men, and patients with metastatic disease have poor outcome even with the most advanced therapeutic approaches. Most cancer therapies target the bulk tumor cells, but may leave intact a small population of tumor‐initiating cells (TICs), which are believed to be responsible for the subsequent relapse and metastasis. Using specific surface markers (CD44, integrin α2β1 and CD133), Hoechst 33342 dye exclusion, and holoclone formation, we isolated TICs from a panel of prostate cancer cell lines (DU145, C4‐2 and LNCaP). We have found that prostate TICs have significant telomerase activity which is inhibited by imetelstat sodium (GRN163L), a new telomerase antagonist that is currently in Phase I/II clinical trials for several hematological and solid tumor malignancies. Prostate TICs telomeres were of similar average length to the telomeres of the main population of cells and significant telomere shortening was detected in prostate TICs as a result of imetelstat treatment. These findings suggest that telomerase inhibition therapy may be able to efficiently target the prostate TICs in addition to the bulk tumor cells, providing new opportunities for combination therapies.
Enhancement of chemotherapeutic agent‐induced apoptosis by inhibition of NF‐κB using ursolic acid
Authors: Li Yunlong , Xing Da , Chen Qun , Chen Wei R.
Published: February 2010

NF‐κB activation is known to reduce the efficiency of chemotherapy in cancer treatment. Ursolic acid, a minimally toxic compound, has shown the capability to inhibit NF‐κB activation in living cells. Here, for the first time, we investigated the effects and mechanisms of NF‐κB inhibition by ursolic acid on chemotherapy treatment (Taxol or cisplatin) of cancer. ASTC‐a‐1 (human lung adenocarcinoma), Hela (human cervical cancer) cells, primary normal mouse cells of lung and liver and mouse in vivo model were used. Activity of signal factors (NF‐κB, Akt, Fas/FasL, BID, Bcl‐2, cytochrome c and caspase‐8, 3) was used to analyze the mechanisms of ursolic acid‐chemo treatment. Ursolic acid‐mediated suppression of NF‐κB drastically reduced the required dosage of the chemotherapeutic agents to achieve identical biological endpoints and enhanced the chemotherapeutic agent‐induced cancer cells apoptosis. Chemosensitization by ursolic acid in cancer cells was dependent on the amplified activation of intrinsic pathway (caspase‐8‐BID‐mitochondria‐cytochrome c‐caspase‐3) by augmentation of BID cleavage and activation of Fas/FasL‐caspase‐8 pathway. Prolonged treatment with relatively low doses of ursolic acid also sensitized cancer cells to the chemotherapeutic agents through suppression of NF‐κB. Chemosensitization by ursolic acid was observed only in cancer cells, but not in primary normal cells. The inhibitive effect of ursolic acid on NF‐κB was reversible, and the reversal was not accompanied by a loss in cells viability. By supplementing chemotherapy with minimally toxic ursolic acid, it is possible to improve the efficacy of cancer treatment by significantly reducing the necessary drug dose without sacrificing the treatment results.
What do we learn from the price of crude oil futures?
Authors: Alquist Ron , Kilian Lutz
Published: February 2010

Despite their widespread use as predictors of the spot price of oil, oil futures prices tend to be less accurate in the mean‐squared prediction error sense than no‐change forecasts. This result is driven by the variability of the futures price about the spot price, as captured by the oil futures spread. This variability can be explained by the marginal convenience yield of oil inventories. Using a two‐country, multi‐period general equilibrium model of the spot and futures markets for crude oil we show that increased uncertainty about future oil supply shortfalls under plausible assumptions causes the spread to decline. Increased uncertainty also causes precautionary demand for oil to increase, resulting in an immediate increase in the real spot price. Thus the negative of the oil futures spread may be viewed as an indicator of fluctuations in the price of crude oil driven by precautionary demand. An empirical analysis of this indicator provides evidence of how shifts in the uncertainty about future oil supply shortfalls affect the real spot price of crude oil. Copyright © 2010 John Wiley & Sons, Ltd.
A control model for tibial cortex neovascularization in the bone chamber
Authors: Winet Howard , Bao Joseph Y. , Moffat Robert
Published: February 2010

Neovascularization across a gap defect in a rabbit tibial cortex was monitored using the optical bone chamber implant (BCI). Cortical bone growing by apposition as trabeculae was observed weekly as it penetrated a slit into a tissue space in vivo and in situ. Each rabbit was viewed weekly with an intravital microscope from 3 to 8 weeks postimplantation. The constant field of view was the slit-gap tissue space, which was 100 μm thick and 2 mm in diameter. Vessels were imaged with epi-illuminated fluorescence microscopy as they carried FITC-dextran 70 that had been injected into an aural vein. Observations were videotaped and photographed. Videotape frames were analyzed with a digital image processing system to obtain measures of vessel length per unit volume (L/V) of fibroblastic granular tissue and trabeculae, caliber C, and flow velocity u, all as functions of time.

Observations supported the conclusions that (1) neovascularization precedes neo-osteogenesis, (2) major vessels tend to align with the tibial axis, (3) bone apposition-generated destruction of fibrous granular tissue vessels stimulates fibrous granular tissue angiogenesis, which keeps its L/V constant, (4) L/V in trabeculae increases with time, and (5) blood supply (Q̄) and nutrient exchange in healing trabeculae are not positively correlated. Thus, O2 supply to the trabeculum cannot be predicted from Q̄ alone because the nutrient exchange area is not constant. It was noted that an increase in the potential nutrient exchange area occurred in both fibrous granular tissue and osseous vessels and the volume fraction of blood decreased in the fibrous granular tissue and remained constant in the trabeculae
Effects of bisphosphonates on isolated Rat osteoclasts as examined by reflected light microscopy
Authors: Sato Masahiko , Grasser William
Published: February 2010

The bisphosphonates 4-amino-1-hydroxybutylidene-1, 1-bisphosphonic acid (ABP), 3-amino-1, 1-hydroxypropane-1, 1-diphosphonic acid (APD), and 1-hyroxyethane-1, 1-diphosphonic acid (EHDP) were compared for their ability to inhibit the osteoclastic resorption of bone in culture. This was achieved by measuring the effect of bisphosphonate concentration on the number of resorption pits formed and the total area of resorption. During this analysis, a new application of reflected polarized light microscopy was developed that has advantages over other microscopy techniques, including scanning electron microscopy (SEM), as applied to the analysis of resorbed bone surfaces. Based on area analysis, the bisphosphonates were effective for the range 10−7–10−8 M, with ABP about two to five times more effective than EHDP or APD. Similar data were obtained by counting the number of resorption pits but with EC50 10 times higher. At lower concentrations (10−9 M), bisphosphonates may enhance the formation of resorption pits. APD was found to be more toxic (10 times) than ABP or EHDP to osteoclasts and mononuclear cells, but toxic concentrations were at least 102 times higher than the resorption EC50. These data plus immunofluorescence, SEM, and transient incubation experiments show that it is the bisphosphonate-bone complex that directly inhibits the excavation of resorption pits by mature osteoclasts. The mechanism of action does not appear to require inhibition of osteoclast differentiation or toxic elimination of osteoclasts. Bisphosphonates, however, subtly affect the cytoskeleton of osteoclasts
Conditioned medium from ras oncogene-transformed nih 3T3 cells induces bone resorption in vitro
Authors: Krieger Nancy S. , Sukhatme Vikas P. , Bushinsky David A. , Sukhatme Vikas P.
Published: February 2010

Tumor-associated hypercalcemia is due, in part, to enhanced osteoclastic bone resorption induced by soluble factors elaborated from malignant cells. rastransformation of NIH 3T3 cells results in a 50-fold induction of cathepsin L mRNA and secretion of the corresponding protein. Since cathepsin L is an acid proteinase we asked whether conditioned medium from these cells would directly increase calcium release from bone in vitro. We tested conditioned medium obtained after 72 h culture of NIH 3T3rar-transformed cells (DT) or nontransformed NIH 3T3 cells (3T3) and identical medium not exposed to cells (Ctl). Incubation of either live or dead neonatal mouse calvaria for 48 h in DT-conditioned medium increased calcium release compared to bones incubated with 3T3 medium. In both states the increased calcium release with DT medium was blocked by 0.25 mM E-64, a general cysteine proteinase inhibitor, and 1 μM Z-Phe-Ala-CH2F, a specific inhibitor of cathepsin L activity. Thus, conditioned medium fromras-transformed cells enhances calcium release in both live and dead bone. Since cathepsin L is the major protein secreted by these cells and the effect of DT-conditioned medium is blocked by a specific inhibitor of cathepsin L, these studies suggest that this acid proteinase acts directly on bone mineral to enhance net calcium release
Cancellous bone remodeling in type i (postmenopausal) osteoporosis: Quantitative assessment of rates of formation, resorption, and bone loss at tissue and cellular levels
Authors: Eriksen Erik F. , Hodgson Stephen F. , Eastell Richard , DR. RIGGS B. Lawrence , Cedel Sandra L. , O'Fallon W. Michael
Published: February 2010

ABSTRACT: The cellular mechanisms for bone loss in type I (postmenopausal) osteoporosis are highly controversial. We attempted to resolve this by assessing rates of formation and resorption of iliac cancellous bone by a new histomorphometric method in 89 women with osteoporosis (mean age + SD, 66 + 6 years) and in 32 carefully selected normal postmenopausal women (64 + 6 years). In the osteoporotic women, bone resorption rate was increased by 39% (P <0.05) at the cellular level and by 67% (P <0.05) at the tissue level, whereas bone formation was unchanged at the tissue level but decreased by 14% (P <0.01) at the cellular (osteoblast) level. This pronounced remodeling imbalance (P <0.001) was probably exacerbated by a 45% increase (P < 0.1) in activation frequency of new remodeling foci. These abnormalities were associated with a high rate of cancellous bone loss (median, 5.8%/year versus 0.1%/year in controls). Thus, accelerated loss of cancellous bone in type I osteoporosis results from the combination of increased bone resorption and inadequate compensation by bone formation.
Bone resorption by isolated osteoclasts in living versus devitalized bone: Differences in mode and extent and the effects of human recombinant tissue inhibitor of metalloproteinases
Authors: Shimizu Hideo , Sakamoto Masako , Sakamoto Seizaburo
Published: February 2010

The incubation of isolated osteoclasts with devitalized bone has become a widely used method for the recent study of bone resorption. Although the studies employing this method have definitively demonstrated that isolated osteoclasts have an avid capacity to resorb devitalized bone, the resorption in this model appears to be different from that of living bone as observed in vivo and in organ culture studies.

To evaluate how the resorption of living bone is different from that of devitalized bone, we have extended this bone resorption model using isolated osteoclasts by including both devitalized and living bone substrates. Living bone substrates were freshly prepared from calvaria of 8- to 12-month-old mice. Periosteum, cellular components, and osteoid were completely scraped off to leave a rigid, smooth, mineral-exposed surface for the isolated osteoclasts to act upon. Some of the bone pieces were devitalized by repeated freezing and thawing. Living and devitalized bones were cultured with isolated rabbit osteoclasts for 60 h with or without recombinant human tissue inhibitor of metalloproteinases (100 μg/ml). The extent of bone resorption was assessed by measuring both the area and the depth of resorption pits.

Comparing the areas of the resorption pits showed significantly more resorption in living bone than in devitalized bone (27% of that of living bone). Recombinant human TIMP reduced the resorption of living bone by 73% but did not, however, inhibit the resorption of devitalized bone. Similarly, resorption pits formed on the living bones were significantly deeper (on the average, 12.4 μm) than those formed on the devitalized bones (on the average, 4.3 μm). The average depth of the resorption pits on living bone was significantly reduced by the presence of the inhibitor, whereas there was no difference between the control and inhibitor-treated devitalized bones. These results suggest that the mechanisms underlying the resorption of living bone and that of devitalized bone are not the same and that the resorption of living bone is aided by osteocytes.
Bone density changes in osteoporosis-prone women exposed to pulsed electromagnetic fields (PEMFs)
Authors: Tabrah Frank , Hoffmeier Mary , Gilbert Fred , Batkin Stanley , Bassett C. A. L.
Published: February 2010

To determine the effect of a 72 Hz pulsating electromagnetic field (PEMF) on bone density of the radii of osteoporosis-prone women, the nondominant forearms of 20 subjects were exposed to PEMF 10 h daily for a period of 12 weeks. Bone density before, during, and after the exposure period was determined by use of a Norland-Cameron bone mineral analyzer. Bone mineral densities of the treated radii measured by single-photon densitometry increased significantly in the immediate area of the field during the exposure period and decreased during the following 36 weeks. A similar but weaker response occurred in the opposite arm, suggesting a “cross-talk” effect on the nontreated radii, from either possible arm proximity during sleep or very weak general field effects. The data suggest that properly applied PEMFs, if scaled for whole-body use, may have clinical application in the prevention and treatment of osteoporosis.
Second messengers in thrombin-stimulated bone resorption
Authors: Dr. Stern Paula H. , Stathopoulos Victoria M. , Shankar Geetha , Fenton John W.
Published: February 2010

Characterized human thrombins and two commercial bovine thrombin preparations were examined for their effects on bone resorption and on the cyclic AMP and phosphoinositide second messenger systems in bone. Human α- and γ-thrombins, as well as both bovine thrombin preparations, stimulated bone resorption in vitro, whereas catalytically inactivated human diisopropylfluorophosphate (DIP)-α-thrombin did not significantly stimulate resorption. Human α-thrombin and a commercial bovine thrombin preparation increased cyclic AMP production in fetal rat limb bones, but another bovine commercial thrombin preparation and γ-thrombin did not. Except for DIP-α-thrombin, all thrombins increased production of inositol phosphates in fetal rat limb bones at concentrations that stimulated resorption. In time course studies, bovine thrombin increased label in inositol trisphosphate at 30 s, with decreasing effects at later times. Inositol monophosphate increased progressively over 30 min. Our results are consistent with thrombin-stimulated bone resorption being mediated at least partially through the inositol phosphate pathway.
Bone mineral content in hypothyroid male patients with hormone replacement: A 3-year study
Authors: Toh Sang H. , Brown Paul H.
Published: February 2010

Several studies have reported that short-term thyroid replacement to the euthyroid state in hypothyroid patients produces loss of both cortical and trabecular bone. However, long-term follow-up studies on this subject have not been reported. We conducted a 3 year follow-up study on 35 white male patients, including 24 patients (groups 1 and 2) with hypothyroidism receiving thyroid hormone replacement to the euthyroid state and 11 patients (groups 3 and 4) with Graves' disease. The bone mineral content of the distal radius was determined by single-photon absorption techniques with an iodine 125 source. It was found that there was no statistically significant difference in bone mineral content by year for any group, indicating no evidence of significant loss of cortical bone mineral by thyroid hormone replacement to the euthyroid state in hypothyroid patients.
Identification of plasminogen activator in osteoclasts
Authors: Grills Brian L. , Allan Elizabeth H. , Yumita Shigeru , Martin T. John , Gallagher James A.
Published: February 2010

Plasminogen activator (PA) was located in newborn rat osteoclasts using a single-cell assay. Immunohistochemistry using biotin-streptavidin-peroxidase indicated the presence of both tissue-type plasminogen activator (tPA) and urokinase (uPA) within the cytoplasm of osteoclasts isolated from newborn rat long bones. Electron microscopic immunohistochemistry using the biotin-streptavidin-colloidal gold system on L.R. Gold thin resin sections of undecalcified, newborn rat tibial metaphyseal trabecular bone identified these proteases in the lysosomal network of osteoclasts. uPA was also localized in marrow macrophage lysosomes, but tPA was not detected in these cells. The localization of these enzymes within osteoclasts may imply their involvement in bone resorption.
Are vitamin D receptor polymorphisms associated with bone mineral density? A meta-analysis
Authors: Cooper Glinda S. , Umbach David M.
Published: February 2010

Vitamin D receptor (VDR) polymorphisms have been strongly associated with bone mineral density (BMD) in some studies but not in others. We used a meta-analytic approach to assess quantitatively the association between VDR and BMD and to examine the influence of specific study characteristics (e.g., skeletal site, mean age of subjects, menopausal status) on the reported results. Sixteen papers published in peer-reviewed journals through July 1996 were included. We calculated the mean difference, percent difference, and effect size (mean difference divided by standard deviation), comparing BMD between homozygous genotypes. At the hip, BMD in the BB genotype was lower than in the bb genotype (mean difference, −0.02 g/cm2; percent difference, −2.4%; and effect size −0.18; p = 0.032). At the spine, the mean difference was −0.03 g/cm2; percent difference, −2.5%; and effect size, −0.19; p = 0.062. At the distal radius, the VDR effect was estimated as the mean difference, −0.01 g/cm2; percent difference, −1.7% and effect size, −0.16; p = 0.078. The spine measurements exhibited the greatest between- and within-study variability. The difference in hip BMD between genotypes was larger (i.e., a more negative number) among the younger women and seemed to decrease with increasing age. However, statistical evidence for this trend was weak (p = 0.06). Data from the spine and the radius showed no evidence of a comparable interaction of the VDR effect with age. When we omitted data from the first report of an association between VDR polymorphisms and BMD, our analyses gave similar results, although the overall effect estimates were smaller. In the combined data from 29 study groups, the BB genotype frequency was 17.2, 4.9, and 2.3% in studies of whites, blacks, and Asians, respectively. VDR polymorphisms represent one genetic factor affecting BMD, but further research into the mechanisms, clinical significance, and its relation between other genetic and environmental factors is needed.
Expression and phorbol ester–induced down-regulation of protein kinase C isozymes in osteoblasts
Authors: Sanders Jennifer L. , Stern Paula H.
Published: February 2010

The protein kinase C (PKC) enzyme family consists of at least 11 isozymes in three classes, with characteristic tissue distributions. Phorbol esters activate and ultimately down-regulate phorbol-sensitive isozymes. PKC is a signal transducer in bone, and phorbol esters influence bone resorption. Little is known about specific PKC isozymes in this tissue, however. We describe here the expression and phorbol ester-induced down-regulation of PKC isozymes in osteoblasts. Normal mouse osteoblasts and seven osteoblastic cell lines (rat UMR-106, ROS 17/2.8, ROS 24/1, and human MG-63, G-292, SaOS-2, HOS-TE85) were screened for isozyme expression by Western immunoblotting using isozyme-specific anti-PKC antibodies. The conventional α and βI, isozymes, but not γ, were present in each of the osteoblasts examined; PKC-βII was detectable in all but the ROS 24/1 line. PKC-ε was expressed in all osteoblasts screened, but other novel PKCs, δ, η, and θ, were detectable only in select lines. The atypical ζ and ι/λ PKCs were in all osteoblasts examined. To determine the sensitivity of the isozymes to prolonged phorbol ester treatment, normal osteoblasts and the UMR-106 cell line were treated with vehicle or 1 μM phorbol 12, 13-dibutyrate (PDB) for 1, 3, 6, 12, 24, or 48 h, and Western blot analysis was performed. Normal and UMR-106 cells showed similar phorbol sensitivities; conventional (α, βI) and novel (δ, ε, η) isozymes were down-regulated by prolonged phorbol treatment but atypical isozymes were not. Down-regulation of all sensitive PKCs was detectable within 6 h of phorbol treatment; the novel δ and ε isozymes, however, showed more rapid and dramatic down-regulation than conventional isozymes. The observed down-regulation was dose-dependent (0.3–3 μM) and specific; 48 h treatment with the inactive phorbol, 4α-phorbol 12,13-didecanoate (4α-PDD), failed to down-regulate PDB-sensitive isozymes. The phorbol-induced down-regulation was also reversible; 24 h after withdrawing PDB, all phorbol-sensitive isozymes, except PKC-η, had recovered at least partially. These studies, the first to characterize thoroughly PKC isozyme expression in osteoblastic cells from several species, demonstrate that osteoblasts have a characteristic PKC isozyme profile, including both phorbol ester–sensitive and –insensitive isozymes. The time course of down-regulation and the presence of phorbol-insensitive PKCs must be considered in interpreting the effects of phorbol esters on bone remodeling.
Osteogenic actions of phenytoin in human bone cells are mediated in part by TGF-β1
Authors: Nakade Osamu , Baylink David J. , Lau K.-H. William
Published: February 2010

We have recently demonstrated that phenytoin, a widely used therapeutic agent for seizure disorders, has osteogenic effects in rats and in humans in vivo, and in human bone cells in vitro. The goal of the present study was to determine the mechanism of the osteogenic action of phenytoin in normal human mandible-derived bone cells. Because many osteogenic agents increased bone cell proliferation through mediation by growth factors, we tested the hypothesis that the osteogenic effects of phenytoin involved the release of a growth factor by measuring the mRNA level of several bone cell growth factors and insulin-like growth factor (IGF) binding proteins with Northern blots using specific cDNA probes. Treatment with 5–50 μM phenytoin reproducibly and markedly increased (up to 6-fold, p < 0.001) the mRNA of transforming growth factor (TGF)-β1, but not that of other growth factors (i.e., IGF-II, platelet-derived growth factor-A [PDGF-A], PDGF-B, and TGF-β2) and IGF binding proteins (i.e., IGFBP-3, −4, and −5). The stimulation was dose dependent, with an optimal dose of 10–50 μM. Maximal increase was seen after 1 h of phenytoin treatment. The release of biologically active TGF-β activity in conditioned media was measured with the mink lung cell proliferation inhibition assay. Twenty-four hours of phenytoin treatment significantly increased the production of biologically active TGF-β (2-fold, p < 0.05) with the optimal dose between 5–50 μM. Comparisons between the in vitro osteogenic effects of phenytoin and those of TGF-β1 reveal that these two agents at their respective optimal doses had similar maximal stimulatory effects on [3H]thymidine incorporation, alkaline phosphatase (ALP)-specific activity, and type Iα-2 collagen mRNA expression in human bone cells. The stimulatory effects of phenytoin on [3H]thymidine incorporation and ALP-specific activity were completely blocked by a neutralizing anti-TGF-β antibody. In conclusion, these findings demonstrate for the first time that at least some of the osteogenic actions of phenytoin in human bone cells could be in part mediated by TGF-β1.
RS-66271, a C-terminally substituted analog of human parathyroid hormone-related protein (1–34), increases trabecular and cortical bone in ovariectomized, osteopenic rats
Authors: Vickery Brian H. , Avnur Zafrira , Cheng Yvonne , Chiou San-San , Leaffer David , Caulfield John P. , Ho Theresa , Krstenansky John L. , Kimmel Donald B.
Published: February 2010

It was predicted from the amino acid sequence of the bone anabolic peptides, parathyroid hormone (PTH) (1–34) and PTH related protein (PTHrP) (1–34), that the C-terminal amino acids form an amphipathic α-helix. Therefore, we substituted a model amphipathic α-helical peptide (MAP) sequence in the C-terminal region of hPTHrP(1–34), obtaining RS-66271 ([MAP1–10]]22–31 hPTHrP(1–34)-NH2). The anabolic activities of RS-66271 and hPTHrP(1–34) were evaluated in 3-month-old, ovariectomized (OVX) osteopenic rats. Subcutaneous injection of hPTHrP(1–34) at 80 μg/kg/day partially reversed estrogen depletion trabecular bone loss but was ineffective in the cortex. In contrast, RS-66271 dose-relatedly reversed loss at both sites and, at 80 μg/kg/day, returned both trabecular and cortical bone calcium to the level of sham-operated controls. Histomorphometric analysis showed significantly elevated bone formation rates over vehicle-treated OVX in both trabecular and cortical tibial bone following treatment with RS-66271. Electron microscopy showed an increase in the relative surface area of vertebral trabeculae covered by osteoblasts in animals treated with RS-66271. These studies demonstrate that the C-terminal amino acids of hPTHrP(1–34) can be replaced by a model amphipathic helix and that the new chemical entity has greater anabolic activity than the parent peptide. The results suggest that RS-66271 may be a candidate molecule for the treatment of human osteoporosis.
The renal clearance of free and conjugated pyridinium cross-links of collagen
Authors: Colwell A. , Eastell R.
Published: February 2010

We developed a sensitive reversed-phase high performance liquid chromatography (HPLC) assay to measure free and total Pyridinoline (Pyr) and Deoxypyridinoline (Dpyr) in serum. The method was used to measure total serum levels in seven premenopausal women (ages 52.2 + 2.4 years) and to investigate the renal clearance of free and conjugated Pyr and Dpyr in two groups of children (group 1: 13 girls, ages 11–13 years; group 2: 18 older children [2 male, 16 female], ages 13–18 years). Blood and 24-h urine samples were collected from the younger group to measure the renal clearance, and blood and 2-h morning urine samples from the older group were collected to investigate the fractional clearance. Total Pyr and Dpyr in the premenopausal women was 4.08 + 0.91 and 1.18 + 0.39 nmol/l, respectively. Free and total Pyr and Dpyr in serum and urine was elevated in both groups of children. The free serum levels were 16 and 18% in young and older children, respectively, compared with 40 and 46% in the urine. The percentage of free Dpyr in serum decreased with total urinary Pyr excretion (r = −0.56, p < 0.005, n = 31). The renal clearance of the free cross-link fraction in both groups was 4-fold higher than the conjugated fraction. The fractional clearance of the free fraction was greater than 1 (p < 0.001) and the conjugated fraction less than 1 (p < 0.001). The fractional excretion of free Dpyr increased with total urinary Pyr excretion (r = 0.66, p < 0.005, n = 13). We conclude that HPLC can be used to measure free and total Pyr and Dpyr in serum and that some free Pyr and Dpyr excreted in urine is produced by the kidney.
Inhibition of HIV‐1 infection by a unique short hairpin RNA to chemokine receptor 5 delivered into macrophages through hematopoietic progenitor cell transduction
Authors: Liang Min , Kamata Masakazu , Chen Kevin N. , Pariente Nonia , An Dong Sung , Chen Irvin S. Y.
Published: February 2010

Background

We recently expressed a potent and noncytotoxic short hairpin (sh)RNA directed against chemokine (c‐c motif) receptor 5 (CCR5) using lentiviral mediated transduction of CD34+ hematopoietic progenitor cells (HPCs) and demonstrated the stable reduction of CCR5 expression in T‐lymphocytes.

Methods

In the present study, we further assessed the activity of the shRNA through HPC transduction and differentiation into macrophages derived from fetal liver CD34+ (FL‐CD34+) HPCs. Transduced lentiviral vector encoding the human CCR5 shRNA was stably maintained in FL‐CD34+ cells and in the terminally differentiated macrophages using macrophage colony‐stimulating factor, granulocyte macrophage colony‐stimulating factor, interleukin‐3 and stem cell factor.

Results

Quantitative real‐time polymerase chain reaction for CCR5 mRNA indicated over 90% reduction of CCR5 mRNA levels in CCR5 shRNA‐transduced population. The cells with knockdown of CCR5 expression acquired resistance to R5 tropic HIV‐1 NFN‐SX strain. We also developed a novel approach utilizing a mCherry‐CCR5 chimeric reporter to assess the effectiveness of CCR5 target down‐regulation in macrophages directly. Both the shRNA and the reporter were maintained throughout HPC differentiation to macrophages without apparent cytotoxicity.

Conclusions

The present study demonstrates a novel method to simply and directly assess the function of small interfering RNA and the effective inhibition of HIV‐1 infection by a potential potent shRNA to CCR5 delivered into macrophages derived from HPCs. Copyright © 2010 John Wiley & Sons, Ltd.
Neoadjuvant therapy for pancreatic cancer: A current review
Authors: Abbott Daniel E. , Baker Marshall S. , Talamonti Mark S.
Published: February 2010

The optimal therapy for pancreatic cancer continues to evolve. Neoadjuvant chemoradiation is a key component of current treatment regimens, and evaluation of previous treatment options will help guide future trials. Here the authors present a review of the current literature with discussion of future directions. J. Surg. Oncol. 2010; 101:315–320. © 2010 Wiley‐Liss, Inc.
Membrane protected conductive polymer as micro‐SPE device for the determination of triazine herbicides in aquatic media
Authors: Bagheri Habib , Khalilian Faezeh , Naderi Mehrnoush , Babanezhad Esmaeil
Published: February 2010

A micro‐SPE technique was developed by fabricating a rather small package including a polypropylene membrane shield containing the appropriate sorbent. The package was used for the extraction of some triazine herbicides from aqueous samples. Solvent desorption was subsequently performed in a microvial and an aliquot of extractant was injected into GC‐MS. Various sorbents including aniline‐ortho‐phenylene diamine copolymer, newly synthesized, polypyrrole, multiwall carbon nanotube, C18 and charcoal were examined as extracting media. Among them, conductive polymers exhibited better performance. Influential parameters including extraction and desorption time, desorption solvent and the ionic strength were optimized. The developed method proved to be rather convenient and offers sufficient sensitivity and good reproducibility. The detection limits of the method under optimized conditions were in the range of 0.01–0.04 ng/mL. The RSDs at a concentration level of 0.1 ng/mL were obtained between 4.5 and 9.3% (n=5). The calibration curves of analytes showed linearity in the range of 0.05–10 ng/mL. The developed method was successfully applied to the extraction of selected triazines from real water samples. The whole procedure showed to be conveniently applicable and quite easy to manipulate.
Enrichment and isolation of barbigerone from Millettia pachycarpa Benth. using high‐speed counter‐current chromatography and preparative HPLC
Authors: Ye Haoyu , Zhong Shijie , Li Yanfang , Tang Minghai , Peng Aihua , Hu Jia , Shi Jie , He Shicao , Wu Wenshuang , Chen Lijuan
Published: February 2010

Enrichment of the anti‐tumor compound barbigerone along with a rotenoid derivative from Millettia pachycarpa Benth. was performed by a two‐step high‐speed counter‐current chromatography (HSCCC) separation process. In the first step, 155.8 mg of target fraction (Fra6) was obtained from 400 mg ethyl acetate extract of M. pachycarpa Benth. with an increase in barbigerone from 5.1 to 13% via HSCCC using a solvent system of n‐hexane–ethyl acetate–methanol–water (5:4:5:3, v/v) under normal phase head to tail elution. HSCCC was repeated to eliminate the major contaminant in this initial fraction 6. After a separation time of 65 min, 22.1 mg barbigerone of 87.7% purity was obtained from Fra6 with the ternary solvent system of n‐hexane–methanol–water (2:2:1, v/v) under normal phase elution. Finally, preparative HPLC was employed for the further isolation of barbigerone and the rotenoid derivative. The structures were confirmed by ESI‐MS, 1H NMR and 13C NMR.
Investigation of commercial sorbents for the analysis of opioid peptides in human plasma by on‐line SPE‐CE
Authors: Benavente Fernando , Medina‐Casanellas Silvia , Barbosa José , Sanz‐Nebot Victoria
Published: February 2010

In this study, we investigated the performance of several commercial sorbents (Sep‐pack® C18, tC18, C8 and tC2, Oasis® HLB, Isolute® ENV+, Strata™‐X and Oasis® MCX) for the determination of opioid peptides by solid‐phase extraction coupled on‐line to capillary electrophoresis (SPE‐CE). First, standard solutions were analyzed in order to achieve the lowest LOD and the best electrophoretic separations using UV detection. The best results were obtained using C18, C8 and tC2 sorbents, which were examined for the analysis of spiked human plasma samples. A double‐step sample clean‐up pretreatment, which consisted of precipitation with acetonitrile and filtration, was needed to prevent saturation of the on‐line SPE microcartridge. The filtration step was critical to obtain optimum analyte recovery and to clean up the sample matrix. A range of centrifugal filters and filtration conditions were tested and the recoveries of the sample pretreatment were evaluated by CE‐ESI‐MS. The LODs attained through SPE‐CE‐UV were approximately ten‐fold better with C18 than with C8 and tC2. The 0.1 μg/mL LODs achieved by C18‐SPE‐CE‐UV were further improved until we could detect 1 ng/mL concentrations of opioid peptides in plasma samples by C18‐SPE‐CE‐ESI‐MS, due to the outstanding selectivity of the MS detection.
Fluoroquinolone antibiotics in environmental waters: Sample preparation and determination
Authors: Speltini Andrea , Sturini Michela , Maraschi Federica , Profumo Antonella
Published: February 2010

The aim of this review is to provide a general overview on the analytical methods proposed in the last decade for trace fluoroquinolone (FQ) determination in environmental waters. A large number of studies have been developed on this topic in reason of the importance of their monitoring in the studies of environmental mobility and potential degradation pathways. Every step of the analysis has been carefully considered, with a particular attention to sample preparation, in relationship with the problems involved in the analysis of real matrices. The different strategies to minimise interference from organic matter and to achieve optimal sensitivity, especially important in those samples with lower FQ concentrations, were also highlighted. Results and progress in this field have been described and critically commented. Moreover, a worldwide overview on the presence of FQs in the environmental waters has been reported.