Triterpene glycosides from the aerial parts of Larrea tridentata. Chemical study of the aerial parts of Larrea tridentata (Zygophyllaceae) resulted in the isolation of 25 triterpene glycosides, 13 of which were previously unknown. Their structures were determined on the basis of comprehensive spectroscopic analyses, including 2D NMR spectroscopy, and hydrolytic cleavage followed by chromatographic and spectroscopic analyses. This is the first systematic phytochemical study of L. tridentata with attention paid to its triterpene glycoside constituents. The isolated compounds were evaluated for their cytotoxic activity against HL-60 human promyelocytic leukemia cells.


Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil. The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15-17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.


Obesity induced changes to plasma adiponectin concentration and cholesterol lipoprotein composition profile in cats. Feline obesity generally results in aberrations to plasma metabolite levels, such as lipid concentrations and lipoprotein composition. This study sought to investigate the resultant effect of obesity on cholesterol lipoprotein composition and circulating adiponectin concentrations in cats. Plasma glucose, lipids (triglyceride, cholesterol and free fatty acid), insulin and adiponectin concentrations, and cholesterol lipoprotein composition were measured and compared between body condition score (BCS) determined normal healthy control and obese cats. Although the obese group demonstrated higher levels of plasma cholesterol, glucose, and triglycerides, as compared to healthy controls, the difference was insignificant thus indicating that the BCS determined obese cats may have been overweight and not morbidly obese. Plasma insulin levels were significantly higher (25-30%) versus healthy control animals thereby possibly hinting at the ensuing emergence of obesity induced insulin resistance. However, the BCS determined obese cat demonstrated a significant reduction (p<0.05) in plasma adiponectin concentration and a significant increase (p<0.05) in LDL-cholesterol % as compared to age matched healthy control animals. This would indicate that changes in plasma adiponectin concentration and cholesterol lipoprotein composition may be good early indicators of obesity in cats.


Efficacy of anthelmintic properties of medicinal plant extracts against Haemonchus contortus. The development of anthelmintic resistance has made the search for alternatives to control gastrointestinal nematodes of small ruminants imperative. Among these alternatives are several medicinal plants traditionally used as anthelmintics. This present work evaluated the efficacy of ethyl acetate, acetone, and methanol dried leaf and seed extracts of five medicinal plants were tested in vitro ovicidal and larvicidal activities on Haemonchus contortus. The in vitro assay was based on egg hatch assay (EHA) and larval development assay (LDA), all plant extracts were evaluated at five concentrations 50, 25, 12.5, 6.25 and 3.13 mg/ml. The leaf and seed ethyl acetate, acetone and methanol of Annona squamosa, Eclipta prostrata, Solanum torvum, Terminalia chebula, and Catharanthus roseus extracts were showed complete inhibition (100%) at the maximum concentration tested (50 mg/ml). The overall findings of the present study have shown that our experimental plant extracts contain possible anthelmintic compounds.


Factors affecting porcine sperm mediated gene transfer. "Sperm mediated gene transfer" (SMGT) is based on the ability of sperm cells to bind exogenous DNA. The main objective of this study was to improve the production of transgenic pigs by SMGT. Taking into account that there is a lack of repeatability in studies of SMGT and that the mechanism of binding and internalization of exogenous DNA is a question that has not been solved, different factors involved in the production of transgenic animals by SMGT method were evaluated. Here we set out to: (1) evaluate the sperm capacity to bind exogenous DNA after DMSO treatment; (2) determine the location of the transgene-spermatozoa interaction; and (3) evaluate the efficiency of production of transgenic piglets by deep intrauterine artificial insemination (AI) with sperm incubated with DNA. The percentage of DNA binding was higher than 30% after 2h of co-culture, but it was not affected by sperm treatment with DMSO (0.3% or 3%). The integrity of the sperm plasma membrane plays a critical role in DNA interaction, and altered plasma membranes facilitate interactions with exogenous DNA. DNA bound mainly to spermatozoa with reduced viability. DNA molecules were found to be mainly associated to the post-acrosomal region (61.9%). After deep intrauterine AI a total of 29 piglets were obtained, but none of them integrated the transgene. In conclusion, although it has been confirmed that DNA can associate with boar spermatozoa, the efficiency of producing transgenic pigs by AI was not confirmed by the present experiments, mainly due to a reduced DNA binding to functional spermatozoa.


In vivo efficacy of tamarind (Tamarindus indica) fruit extract on experimental fluoride exposure in rats. The study was undertaken to determine the efficacy of hydro-methanolic (1:1) extract of tamarind (Tamarindus indica L.) fruit pulp in removing body fluoride burden. Thirty rats were divided into five groups. Keeping no fluoride group as the control, rats of no treatment, low dose, middle dose and high dose groups received sodium fluoride orally at the rate of 200mg per kg body weight daily for 14 weeks. Rats of low dose, middle dose and high dose group simultaneously received tamarind fruit pulp extract at three doses, viz. 25 (low), 50 (medium) and 100mg (high) per kg body weight orally, respectively. Fluoride concentration in blood, urine and long bone of experimental rats was monitored to assess the efficacy of the extract. Mean serum fluoride concentration in fluoride exposed rats was 0.145 ± 0.009 and 0.783 ± 0.042 μg/ml on days 0 and 98. In comparison, fluoride concentrations in tamarind treated rats were 0.179 ± 0.021 and 0.633 ± 0.015; 0.179 ± 0.021 and 0.502 ± 0.025 and 0.176 ± 0.021 and 0.498 ± 0.030 μg/ml in low, medium and high dose groups, respectively on day 0 and day 98 of the experiment. There was a significant (p ≤ 0.01) increase in urinary fluoride excretion from day 28 onwards. The mean fluoride concentration in long bones of treated rats was significantly lower than the values recorded in fluoride exposed rats. These findings suggest that concomitant use of tamarind fruit pulp extract can reduce fluoride concentration in blood and bone and enhanced urinary excretion, indicating the ameliorative potential of fruits of tamarind in fluoride toxicity.


Low-cost struvite production using source-separated urine in Nepal. This research investigated the possibility of transferring phosphorus from human urine into a concentrated form that can be used as fertilizer in agriculture. The community of Siddhipur in Nepal was chosen as a research site, because there is a strong presence and acceptance of the urine-diverting dry toilets needed to collect urine separately at the source. Furthermore, because the mainly agricultural country is landlocked and depends on expensive, imported fertilizers, the need for nutrient security is high. We found that struvite (MgNH(4)PO(4)·6H(2)O) precipitation from urine is an efficient and simple approach to produce a granulated phosphorus fertilizer. Bittern, a waste stream from salt production, is a practical magnesium source for struvite production, but it has to be imported from India. Calculations show that magnesium oxide produced from locally available magnesite would be a cheaper magnesium source. A reactor with an external filtration system was capable of removing over 90% of phosphorus with a low magnesium dosage (1.1 mol Mg mol P), with coarse nylon filters (pore width up to 160±50 μm) and with only one hour total treatment time. A second reactor setup based on sedimentation only achieved 50% phosphate removal, even when flocculants were added. Given the current fertilizer prices, high volumes of urine must be processed, if struvite recovery should be financially sustainable. Therefore, it is important to optimize the process. Our calculations showed that collecting the struvite and calcium phosphate precipitated spontaneously due to urea hydrolysis could increase the overall phosphate recovery by at least 40%. The magnesium dosage can be optimized by estimating the phosphate concentration by measuring electrical conductivity. An important source of additional revenue could be the effluent of the struvite reactor. Further research should be aimed at finding methods and technologies to recover the nutrients from the effluent.


The influence of gill and liver metabolism on the predicted bioconcentration of three pharmaceuticals in fish. The potential for xenobiotic compounds to bioconcentrate is typically expressed through the bioconcentration factor (BCF), which has gained increased regulatory significance over the past decade. Due to the expense of in vivo bioconcentration studies and the growing regulatory need to assess bioconcentration potential, BCF is often calculated via single-compartment models, using K(OW) as the primary input. Recent efforts to refine BCF models have focused on physiological factors, including the ability of the organism to eliminate the compound through metabolic transformation. This study looks at the ability of in vitro biotransformation assays using S9 fractions to provide an indication of metabolic potential. Given the importance of the fish gill and liver in metabolic transformation, the metabolic loss of ibuprofen, norethindrone and propranolol was measured using rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) gill and liver S9 fractions. Metabolic transformation rates (k(M)) were calculated and integrated into a refined BCF model. A significant difference was noted between BCF solely based on K(OW) and BCF including k(M). These studies indicate that the inclusion of k(M) in BCF models can bring predicted bioconcentration estimates closer to in vivo values.


Interaction of engineered nanoparticles with various components of the environment and possible strategies for their risk assessment. Nanoparticles are the materials with at least two dimensions between 1 and 100 nm. Mostly these nanoparticles are natural products but their tremendous commercial use has boosted the artificial synthesis of these particles (engineered nanoparticles). Accelerated production and use of these engineered nanoparticles may cause their release in the environment and facilitate the frequent interactions with biotic and abiotic components of the ecosystems. Despite remarkable commercial benefits, their presence in the nature may cause hazardous biological effects. Therefore, detail understanding of their sources, release interaction with environment, and possible risk assessment would provide a basis for safer use of engineered nanoparticles with minimal or no hazardous impact on environment. Keeping all these points in mind the present review provides updated information on various aspects, e.g. sources, different types, synthesis, interaction with environment, possible strategies for risk management of engineered nanoparticles.


Alkylphenol oxidation with a laccase from a white-rot fungus: effects of culture induction and of ABTS used as a mediator. We investigated the potential of the laccase from the white-rot fungus Marasmius quercophilus to transform certain alkylphenols (p-nonylphenol, p-octylphenol and p-t-octylphenol). We tested the reactivity of this enzyme under different conditions: in liquid cultures and using the partially purified laccase with and without 2,2'-azino-bis-3-ehtylbenzothiazoline-6-sulfonicacid (ABTS) as a mediator. The percentage of p-t-octylphenol disappearance in liquid cultures was 69.0 ± 1.5% and 81 ± 5% after a 8-d or 15-d incubation, respectively, with p-nonylphenol, these percentages were 62 ± 4% and 91 ± 6% and with p-octylphenol 37 ± 3% and 65 ± 1% after a 15-d and a 21-d incubations, respectively. Induced pre-cultures were also used to inoculate the liquid cultures to enhance p-octylphenol transformation: the percentages of disappearance were 91.0 ± 0.5% and 97 ± 1% after a 8-d and a 15-d incubation, respectively. Mass spectrometry analysis showed that the products of oxidation of p-octylphenol were dimers with a mass of 411 m/z. Furthermore, we identified a purple compound (m/z 476) formed when ABTS was added to the reaction medium with the purified laccase. This result confirms that, in complex environments such as soils or litters where many molecules can interact with the enzyme substrate or the product of oxidation, laccase activities and those of other phenoloxidases should not be measured with ABTS.


Secretion profiles of fungi as potential tools for metal ecotoxicity assessment: a study of enzymatic system in Trametes versicolor. The relationship between the expression of extracellular enzymatic system and a metal stress is scarce in fungi, hence limiting the possible use of secretion profiles as tools for metal ecotoxicity assessment. In the present study, we investigated the effect of Zn, Cu, Pb and Cd, tested alone or in equimolar cocktail, on the secretion profiles at enzymatic and protein levels in Trametesversicolor. For that purpose, extracellular hydrolases (acid phosphatase, β-glucosidase, β-galactosidase and N-acetyl-β-glucosaminidase) and ligninolytic oxidases (laccase, Mn-peroxidase) were monitored in liquid cultures. Fungal secretome was analyzed by electrophoresis and laccase secretion was characterized by western-blot and mass spectrometry analyses. Our results showed that all hydrolase activities were inhibited by the metals tested alone or in cocktail, whereas oxidase activities were specifically stimulated by Cu, Cd and metal cocktail. At protein level, metal exposure modified the electrophoretic profiles of fungal secretome and affected the diversity of secreted proteins. Two laccase isoenzymes, LacA and LacB, identified by mass spectrometry were differentially glycosylated according to the metal exposure. The amount of secreted LacA and LacB was strongly correlated with the stimulation of laccase activity by Cu, Cd and metal cocktail. These modifications of extracellular enzymatic system suggest that fungal oxidases could be used as biomarkers of metal exposure.


Effects of selected pharmaceutically active compounds on the ammonia oxidizing bacterium Nitrosomonas europaea. Pharmaceutically active compounds (PhACs) are commonly found in wastewater influent. However, little research has focused on determining their impact on fundamental processes in wastewater treatment such as nitrogen removal. In this study, focus was placed on 4 commonly occurring PhACs (ketoprofen, naproxen, carbamazepine and gemfibrozil). Their effect was ascertained in the ammonia oxidizing bacterium (AOB), Nitrosomonas europaea in terms of membrane integrity and nitrite production. These PhACs were shown to inhibit nitrite production at concentrations of 1 and 10 μM while no effect was observed at 0.1 μM. The maximum observed nitrification inhibition was 25%, 29%, 22% and 26% for ketoprofen, naproxen, carbamazepine and gemfibrozil, respectively. A decrease in the live/dead ratio ranging from 10% to 16% suggests that these PhACs affect membrane integrity in N.europaea. The difference in nitrite production between PhACs treated cells and non PhAC treated controls was still significant following washing suggesting that inhibition is irreversible. Finally, nitrite production when adjusted to the live fraction of cells was also found to decrease suggesting that PhACs inhibited the activity of surviving cells. These results suggest that the presence of PhACs may affect AOB activity and may impact nitrogen removal, a key function in wastewater treatment. Follow up studies with additional AOB and in mixed culture are needed to further confirm these results.


Levels and congener profiles of PCBs and PCDD/Fs in blue shark (Prionace glauca) liver from the South-Eastern Mediterranean Sea (Italy). Liver of blue shark (Prionace glauca) specimens from the South-Eastern Mediterranean Sea were analyzed for the presence of polychlorinated biphenyls (PCBs), including coplanar congeners, polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs). PCBs were the dominant chemicals, followed by PCDFs and PCDDs. The pattern of PCB congener concentrations in the hepatic tissue was dominated by higher chlorinated compounds. The specific profile of toxic PCDD/F congeners was characterized mainly by 2,3,7,8-TCDF and 2,3,7,8-TCDD, followed by 1,2,3,6,7,8-HxCDD and 2,3,4,6,7,8-HxCDF. The total 2,3,7,8-TCDD toxic equivalent (TEQs) was 149 pg g⁻¹ lipid wt. The profile of TEQ shows that PCDDs present the greatest risk to this species contributing to total toxicity with a percentage approximately of 60%, while the contribution of PCDFs and DL-PCBs is almost the same being 22.4% and 21.6%, respectively. Further investigations are urgently needed to characterize the PCDD/Fs contamination levels not only in elasmobranch fish but in all Mediterranean marine biota.


Removal of dissolved estrogen in sewage effluents by β-cyclodextrin polymer. Substances with estrogenic activity are found in effluents of municipal sewage plants and dairy farms. These effluents have the potential to induce feminization in male fish. In this study, cyclodextrin polymers (CDPs) that are insoluble in both polar and non-polar solvents were selected for the removal of dissolved estrogens in the effluent of a municipal sewage plant. The removal capacity of CDPs was high in the order of β-CDP≥γ-CDP≫α-CDP. The mechanism for adsorption of estrogens to β-CDP was not only due to a host-guest interaction as molecular recognition by β-cyclodextrin (β-CD), but also due to adsorption by the polymer matrix. β-CDP of 0.2% (w/v) removed 17β-estradiol (E2) of about 70% from 10(-11)mol/L, and more than 90% from ≥10(-10)mol/L. The removal ratios of E2 in the presence of cholesterols, which are contained at higher concentrations than estrogens in sewage effluents and are adsorptive competitor for β-CDP, were about 85% at a cholesterol/E2 molar ratio of 100 and >90% at molar ratios of 0.1, 1, and 10. The effluent from a municipal sewage plant had estrogenic activity corresponding to 5.5×10(-11)molE2/L by yeast two-hybrid assay. The estrogens in the effluent were also removed >90% by the β-CDP treatment. Therefore, β-CDP is able to remove dissolved estrogens over a wide range of concentrations in the presence of various contaminants such as wastewaters.


Novel robotic da Vinci instruments for laparoendoscopic single-site surgery. To describe novel robotic laparoendoscopic single-site surgery (R-LESS) instruments, and present the initial laboratory experience in urology. The VeSPA surgical instruments (Intuitive Surgical, Sunnyvale, CA) were designed to be used with the DaVinci Si surgical system. A multichannel port and curved cannulae were inserted through a single 3.5-cm umbilical incision. The port allowed 1 scope, 2 robotic instruments, and a 5- to 12-mm assistant instrument. Four pyeloplasties (right 2, left 2), 4 partial nephrectomies (right 2, left 2), and 8 nephrectomies (right 4, left 4) were performed in 4 female farm pigs (mean weight, 34.5 kg). Technical feasibility and efficiency were assessed in addition to perioperative outcomes. All 16 R-LESS procedures were performed successfully without the addition of laparoscopic ports or open conversion. Mean total operative time was 110 minutes (range, 82-127), and mean blood loss was 20 mL (range, 10-100). Mean warm ischemia time for partial nephrectomy was 14.8 minutes (range, 12-20). There were no intraoperative complications. No robotic system failures occurred, and robotic instrument clashing was found to be minimal. One needle driver malfunctioned and assistant movement was limited. R-LESS kidney surgery using the VeSPA instruments is feasible and efficient in the porcine model. The system offers a wide range of motion, instrument and scope stability, improved ergonomics, and minimal instrument clashing. Although preliminary experience is encouraging, further refinements are expected to optimize urological applications of this robotic technology.


Poliomyelitis eradication: another step forward.


Competitive time- and density-dependent adhesion of staphylococci and osteoblasts on crosslinked poly(ethylene glycol)-based polymer coatings in co-culture flow chambers. Biomaterial-associated infections (BAI) remain a serious clinical complication, often arising from an inability of host tissue-implant integration to out-compete bacterial adhesion and growth. A commercial polymer coating based on polyethylene glycol (PEG), available in both chemically inert and NHS-activated forms (OptiChem(®)), was compared for simultaneous growth of staphylococci and osteoblasts. In the absence of staphylococci, osteoblasts adhered and proliferated well on glass controls and on the NHS-reactive PEG-based coating over 48 h, but not on the inert PEG coating. Staphylococcal growth was low on both PEG-based coatings. When staphylococci were pre-adhered on surfaces for 1.5 h to mimic peri-operative contamination, osteoblast growth and spreading was reduced on glass but virtually absent on both reactive and inert PEG-based coatings. Thus although NHS-reactive, PEG-based coatings stimulated tissue-cell interactions in the absence of contaminating staphylococci, the presence of adhering staphylococci eliminated osteoblast adhesion advantages on the PEG surface. This study demonstrates the importance of using bacterial and cellular co-cultures compared to monocultures when assessing functionalized biomaterials coatings for infectious potential.


Immunogenicity of bivalent types 1 and 3 oral poliovirus vaccine: a randomised, double-blind, controlled trial. Poliovirus types 1 and 3 co-circulate in poliomyelitis-endemic countries. We aimed to assess the immunogenicity of a novel bivalent types 1 and 3 oral poliovirus vaccine (bOPV). We did a randomised, double-blind, controlled trial to assess the superiority of monovalent type 2 OPV (mOPV2), mOPV3, or bOPV over trivalent OPV (tOPV), and the non-inferiority of bivalent vaccine compared with mOPV1 and mOPV3. The study was done at three centres in India between Aug 6, 2008, and Dec 26, 2008. Random allocation was done by permuted blocks of ten. The primary outcome was seroconversion after one monovalent or bivalent vaccine dose compared with a dose of trivalent vaccine at birth. The secondary endpoints were seroconversion after two vaccine doses compared with after two trivalent vaccine doses and cumulative two-dose seroconversion. Parents or guardians and study investigators were masked to treatment allocation. Because of multiple comparisons, we defined p≤0·01 as statistically significant. This trial is registered with Current Controlled Trials, ISRCTN 64725429. 900 newborn babies were randomly assigned to one of five vaccine groups (about 180 patients per group); of these 70 (8%) discontinued, leaving 830 (92%) for analysis. After the first dose, seroconversion to poliovirus type 1 was 20% for both mOPV1 (33 of 168) and bOPV (32 of 159) compared with 15% for tOPV (25 of 168; p>0·01), to poliovirus type 2 was 21% (35 of 170) for mOPV2 compared with 25% (42 of 168) for tOPV (p>0·01), and to poliovirus type 3 was 12% (20 of 165) for mOPV3 and 7% (11 of 159) for bOPV compared with 4% (7 of 168) for tOPV (mOPV3 vs tOPV p=0·01; bOPV vs tOPV; p>0·01). Cumulative two-dose seroconversion to poliovirus type 1 was 90% (151 of 168) for mOPV1 and 86% (136 of 159) for bOPV compared with 63% (106 of 168) for tOPV (p<0·0001), to poliovirus type 2 was 90% (153 of 170) for mOPV2 compared with 91% (153 of 168) for tOPV (p>0·01), and to poliovirus type 3 was 84% (138 of 165) for mOPV3 and 74% (117 of 159) for bOPV compared with 52% (87 of 168) for tOPV (p<0·0001). The vaccines were well tolerated. 19 serious adverse events occurred, including one death; however, these events were not attributed to the trial interventions. The findings show the superiority of bOPV compared with tOPV, and the non-inferiority of bOPV compared with mOPV1 and mOPV3. GAVI Alliance, World Health Organization, and Panacea Biotec.


Chitosan/poly(epsilon-caprolactone) blend scaffolds for cartilage repair. Chitosan (CHT)/poly(ɛ-caprolactone) (PCL) blend 3D fiber-mesh scaffolds were studied as possible support structures for articular cartilage tissue (ACT) repair. Micro-fibers were obtained by wet-spinning of three different polymeric solutions: 100:0 (100CHT), 75:25 (75CHT) and 50:50 (50CHT) wt.% CHT/PCL, using a common solvent solution of 100 vol.% of formic acid. Scanning electron microscopy (SEM) analysis showed a homogeneous surface distribution of PCL. PCL was well dispersed throughout the CHT phase as analyzed by differential scanning calorimetry and Fourier transform infrared spectroscopy. The fibers were folded into cylindrical moulds and underwent a thermal treatment to obtain the scaffolds. μCT analysis revealed an adequate porosity, pore size and interconnectivity for tissue engineering applications. The PCL component led to a higher fiber surface roughness, decreased the scaffolds swelling ratio and increased their compressive mechanical properties. Biological assays were performed after culturing bovine articular chondrocytes up to 21 days. SEM analysis, live-dead and metabolic activity assays showed that cells attached, proliferated, and were metabolically active over all scaffolds formulations. Cartilaginous extracellular matrix (ECM) formation was observed in all formulations. The 75CHT scaffolds supported the most neo-cartilage formation, as demonstrated by an increase in glycosaminoglycan production. In contrast to 100CHT scaffolds, ECM was homogenously deposited on the 75CHT and 50CHT scaffolds. Although mechanical properties of the 50CHT scaffold were better, the 75CHT scaffold facilitated better neo-cartilage formation.


Biocompatibility and osteogenesis of biomimetic Bioglass-Collagen-Phosphatidylserine composite scaffolds for bone tissue engineering. A novel biomimetic composite scaffold Bioglass-Collagen-Phosphatidylserine (BG-COL-PS) was fabricated with a freeze-drying technique. The macrostructure and morphology as well as mechanical strength of the scaffolds were characterized. Scanning electronic microscopy (SEM) showed that the BG-COL-PS scaffolds exhibited interconnected porous structures with pore sizes of several microns up to about 300 μm. The scaffolds have a porosity of 75.40% and the corresponding compressive strength of 1.5469 Mpa. Rat mesenchymal stem cells (rMSCs) were seeded on BG-COL-PS or BG-COL scaffolds and cultured for 21 days in vitro. Based on the results of SEM, dsDNA content, alkaline phosphatase (ALP) activity, osteogenic gene expression analysis and alizarin red staining, the responses of MSCs to the scaffold exhibited a higher degree of attachment, growth as well as osteogenic differentiation than those on BG-COL scaffolds in vitro. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both pure BG-COL-PS scaffolds and MSC/scaffold constructs were implanted in rat femurs defects for 6 weeks and studied histologically and radiographically. The in vivo results showed that BG-COL-PS composite scaffolds exhibited good biocompatibility and extensive osteoconductivity with host bone. Moreover, the BG-COL-PS/MSC constructs dramatically enhanced the efficiency of new bone formation than pure BG-COL-PS scaffolds or BG-COL/MSC constructs. All these results demonstrate the usefulness of PS composited BG-COL-PS scaffolds for inducing enhanced bone formation. The BG-COL-PS scaffolds fulfill the basic requirements of bone tissue engineering scaffold and have the potential to be applied in orthopedic and reconstructive surgery.


RANKL-induced TRPV2 expression regulates osteoclastogenesis via calcium oscillations. The receptor activator of NFκB ligand (RANKL) induces Ca(2+) oscillations and activates the Nuclear Factor of Activated T cells 1 (NFATc1) during osteoclast differentiation (osteoclastogenesis). Ca(2+) oscillations are an important trigger signal for osteoclastogenesis, however the molecular basis of Ca(2+) permeable influx pathways serving Ca(2+) oscillations has not yet been identified. Using a DNA microarray, we found that Transient Receptor Potential Vanilloid channels 2 (TRPV2) are expressed significantly in RANKL-treated RAW264.7 cells (preosteoclasts) compared to untreated cells. Therefore, we further investigated the expression and functional role of TRPV2 on Ca(2+) oscillations and osteoclastogenesis. We found that RANKL dominantly up-regulates TRPV2 expression in preosteoclasts, and evokes spontaneous Ca(2+) oscillations and a transient inward cation current in a time-dependent manner. TRPV inhibitor ruthenium red and tetracycline-induced TRPV2 silencing significantly decreased both the frequency of Ca(2+) oscillations and the transient inward currents in RANKL-treated preosteoclasts. Silencing of store-operated Ca(2+) entry (SOCE) proteins similarly suppressed both RANKL-induced oscillations and currents in preosteoclasts. Furthermore, suppression of TRPV2 also reduced RANKL-induced NAFTc1 expression, its nuclear translocation, and osteoclastogenesis. In summary, Ca(2+) oscillations in preosteoclasts are triggered by RANKL-dependent TRPV2 and SOCE activation and intracellular Ca(2+) release. Subsequent activation of NFATc1 promotes osteoclastogenesis.


Novel variant form of t(11;22)(q23;q13)/MLL-EP300 fusion transcript in the evolution of an acute myeloid leukemia with myelodysplasia-related changes.


Volunteer and user evaluation of the National Sexual Assault Online Hotline. The National Sexual Assault Online Hotline (NSAOH) is a new model for delivery of rape and sexual assault crisis services through a secure, confidential chat-based online hotline. This paper presents a program evaluation drawn from volunteer counselor and user perceptions and experiences during the second year of operation of the NSAOH. Outcome data are presented from 731 session evaluations submitted by 94 volunteers and session evaluations from 4609 user sessions collected between June 1, 2008 and May 30, 2009. Evaluation includes ratings of usefulness, topics discussed, length of sessions, services provided, and session difficulties. The results indicate that the model is viable and useful, and the majority of volunteers and users are satisfied. Volunteer knowledge and skills are strongly associated with satisfaction with the hotline. Nevertheless, one-fifth of volunteers rate their session as not useful and users rate 8.2% of volunteers low in knowledge and skills. NSAOH is reaching many who have not previously sought services or did not resolve issues through other means. Findings suggest the importance of preparing volunteers in both crisis intervention and a wide variety of long-term issues related to sexual assault. Recommendations for program development, evaluation, and further research are presented.


Development of microbial engineered whole-cell systems for environmental benzene determination. This paper reports the development of two recombinant bacterial systems that can be used to monitor environmental benzene contamination based on Escherichia coli, which carry genes coding for benzene dioxygenase and benzene dihydrodiol dehydrogenase from Pseudomonas putida MST. E. coli strains express these two enzymes under the control of the Ptac promoter or without any induction. These activities can be detected electrochemically or colorimetrically and used to monitor benzene pollution in environmental air samples collected from an oil refinery assessing benzene by different laboratory experimental procedures. The procedures involving whole-cell bioassays determine the concentration of benzene through benzene dioxygenase activity, which allows for direct correlation of oxygen consumption, and through the benzene dihydrodiol dehydrogenase that causes catechol accumulation and restores NADH necessary for the activity of the first enzyme. Oxygen consumption and catechol production deriving from both enzymatic activities are related to benzene concentration and their measurements determined the sensitivity of the system. The results indicated that the sensitivity was enough to detect the benzene vapor at a lower concentration level of 0.01 mM in about 30 min. The possibility for on-line monitoring of benzene concentration by our new recombinant cells results from the fact that no particular treatment of environmental samples is required. This is a major advantage over other biosensors or assays. Moreover, the development of microbial cells that did not require any addition or effectors for the transcription of the specific enzymes, allowed these systems to be more versatile in automated environmental benzene monitoring.


Testing sex and gender in sports; reinventing, reimagining and reconstructing histories. Most international sports organisations work on the premise that human beings come in one of two genders: male or female. Consequently, all athletes, including intersex and transgender individuals, must be assigned to compete in one or other category. Since the 1930s (not, as is popularly suggested, the 1960s) these organisations have relied on scientific and medical professionals to provide an 'objective' judgement of an athlete's eligibility to compete in women's national and international sporting events. The changing nature of these judgements reflects a great deal about our cultural, social and national prejudices, while the matter of testing itself has become a site of conflict for feminists and human rights activists. Because of the sensitive nature of this subject, histories of sex testing are difficult to write and research; this has lead to the repetition of inaccurate information and false assertions about gender fraud, particularly in relation to the 'classic' cases of Stella Walsh and Heinrich/Hermann/Dora Ratjen. As historians, we need to be extremely careful to differentiate between mythologies and histories.


Cytokine induction by immunostimulatory DNA in porcine PBMC is impaired by a hairpin forming sequence motif from the genome of Porcine Circovirus type 2 (PCV2). Porcine Circovirus type 2 (PCV2) can cause postweaning multisystemic wasting syndrome (PMWS) in young pigs with severe immunosuppression as a major characteristic of the disease complex. Despite the dramatic involvement of the immune system, the interaction between PCV2 and the host is until date not well understood. The DNA genome of PCV2 contains sequences that in synthetic form (oligodeoxyribonucleotides; ODNs) can act immunomodulatory on porcine peripheral blood mononuclear cells (poPBMCs) in vitro. One such sequence (ODN PCV2/1) acts inhibitory on interferon (IFN)-α production induced by immunostimulatory DNA but not that induced by RNA, and the inhibitory activity is dependent on secondary structure formation. In the present study, the characteristic of ODN PCV2/1 was examined further by altering the nucleotide sequence to disrupt hairpin structure formation but still enable multimer structures through G-tetrads. This modification resulted in loss of IFN-α-inhibitory activity of the ODN and thus indicated the importance of hairpin structures. In addition, ODN PCV2/1 was compared to another inhibitory ODN (IRS 869) previously used in human and murine cells. In contrast to ODN PCV2/1, ODN IRS 869 did not inhibit IFN-α production induced by class A ODN 2216 but was a more efficient inhibitor of IFN-α production induced by plasmid DNA than ODN PCV2/1. In cultures induced by the RNA stimulator Poly I:C, however, a strong synergistic IFN-α stimulatory effect was seen in combination with ODN IRS 869. These results indicate that ODN PCV2/1 and ODN IRS 869 function through separate mechanisms to affect cytokine production by immune cells. The effect of ODN PCV2/1 was studied further by monitoring the expression of mRNA for IFN-α, IL-12p40, IL-10, IL-6, IFN-γ, IL-1β, TGF-β, and TNF-α in cultures of poPBMC stimulated with ODN 2216 or Poly I:C. Results from qPCR analyses showed that ODN PCV2/1 clearly inhibited the expression of IFN-α, IL-12p40, IL-10 and IL-6 when induced by ODN 2216, but did not seem to affect any of the cytokines examined when induced by Poly I:C. Initial studies using confocal microscopy and fluorochrome labelled ODNs indicate that ODN 2216 and ODN PCV2/1 co-localize in subpopulations of poPBMC.


Expression of four canine leukocyte adhesion factors in fresh and stored whole blood samples evaluated using a no-lyse, no-wash method. Expression of four leukocyte adhesion factors on canine leukocytes was studied by flow cytometry using a no-lyse, no-wash method. The effect of fixation and storage for up to 14 days in 1% paraformaldehyde on labelled samples and within assay variation was evaluated. Monoclonal antibodies directed to monocyte marker CD14, and to adhesion molecules CD11a, CD18, CD32 and CD49d were used. Cell surface marker, cell population, time, and the interactions between time and cell marker significantly affected expression of cell adhesion factors. For CD18, there was a significant difference in mean fluorescence intensity (MFI) between fresh and stored samples (P<0.001), but no significant difference between stored samples. The MFIs of CD11a and CD49d were not significantly affected by fixation and storage. The CVs differed significantly depending on cell marker (P<0.001) and cell population (P=0.005). Fixation and storage did not significantly affect the CV. In conclusion, a no-lyse, no-wash method can be applied to canine leukocytes. The effect of fixation and storage on the resulting MFI differs between monoclonal antibodies, and should be evaluated for each antibody before use. The coefficient of variation was generally acceptable, and high CVs were related to a low MFIs or low numbers of analysed cells.


In vitro effect of different mediators of apoptosis on canine cranial and caudal cruciate ligament fibroblasts and its reversibility by pancaspase inhibitor zVAD.fmk. Primary fibroblast cultures of canine cranial (CCL) and caudal (CaCL) cruciate ligaments were stimulated with different apoptosis inducers with or without preincubation of the pancaspase inhibitor zVAD.fmk. In contrast to CaCL fibroblasts, fibroblasts from CCL were significantly more susceptible to apoptosis inducers of the intrinsic pathway like doxorubicin, cisplatin and nitric oxide (NO)-donors and to Fas ligand (FasL), an apoptosis inducer of the death receptor pathway. Apoptotic response to staurosporine and the peroxynitrite donor GEA was similar in both ligament fibroblasts. Stimulation with dexamethasone or TNFα could not induce apoptosis in CCL and CaCL fibroblasts, in spite of present TNFR1 and TNFR2 receptors. zVAD.fmk was able to prevent apoptosis in up to 66% of CCL cells when treated with FasL, cisplatin or doxorubicin but it had no effect on NO or peroxynitrite induced apoptosis. In conclusion, differential susceptibility to apoptotic triggers like FasL or NO between cranial and caudal cruciate ligament fibroblasts in vitro may be a reflection of the different susceptibilities to degenerative rupture of the ligament. These findings indicate that a general caspase inhibition does not completely protect canine CCL cells from apoptosis.


Endoscopic anterior cricoid split and balloon dilation in pediatric subglottic stenosis. To analyze the outcome of a new endoscopic approach for the treatment of pediatric subglottic stenosis. Case series. Tertiary care center. Eighteen pediatric cases of grade II to IV subglottic stenosis (8 congenital and 10 acquired) consecutively treated at our institutions by Endoscopic Anterior Cricoid Split (EACS) and balloon dilation between 2006 and 2010. Treatment protocol encompassed systematic postoperative laryngeal stenting (7 days of intubation or 1 month of Montgomery T-tube in previously tracheotomized patients) and endoscopic controls with possible additional balloon dilation every 15 days for at least 2 months. Patients' ages ranged from 1 to 101 months. Postoperative follow-up ranged from 4 to 45 months (median value±SD: 15.3±11.9). The mean duration of the endoscopic procedure was 35.2±13.2 min. The number of days spent in PICU during the perioperative period varied between 2 and 15. Four patients (22.2%) needed one and 14 patients (77.7%) required several (from 4 to 7) additional balloon dilations during the postoperative endoscopic controls. No incident was observed during or immediately after EACS. Treatment was efficient in 83% of cases (n=15), with no residual respiratory symptoms and grade 0 to 1 SGS at the end of follow-up. EACS is a safe and efficient technique to treat pediatric subglottic stenosis, regardless of their grade and length, provided to associate it with postoperative laryngeal stenting and regular endoscopic follow-up with possible additional balloon dilations. In our teams, it has become the first line treatment for most grades II to IV SGS. Its indications can be extended to congenital stenosis with cartilaginous involvement and to long-lasting acquired stenosis with firm fibrosis.


A testable theory of problem solving courts: Avoiding past empirical and legal failures. Recent years have seen a proliferation of problem solving courts designed to rehabilitate certain classes of offenders and thereby resolve the underlying problems that led to their court involvement in the first place. Some commentators have reacted positively to these courts, considering them an extension of the philosophy and logic of Therapeutic Jurisprudence, but others show concern that the discourse surrounding these specialty courts has not examined their process or outcomes critically enough. This paper examines that criticism from historical and social scientific perspectives. The analysis culminates in a model that describes how offenders are likely to respond to the process as they engage in problem solving court programs and the ways in which those courts might impact subsequent offender conduct. This Therapeutic Jurisprudence model of problem solving courts draws heavily on social cognitive psychology and more specifically on theories of procedural justice, motivation, and anticipated emotion to offer an explanation of how offenders respond to these programs. We offer this model as a lens through which social scientists can begin to address the concern that there is not enough critical analysis of the process and outcome of these courts. Applying this model to specialty courts constitutes an important step in critically examining the contribution of problem solving courts.


TLR-9, NOD-1, NOD-2, RIG-I and immunoglobulins in recurrent otitis media with effusion. Pattern recognition receptors (PRRs) induce appropriate immune responses after recognizing certain molecular characteristics of pathogens. It is not known, however, whether PRRs are expressed in middle ear infections and whether the expression of PRRs and immunoglobulins is correlated in recurrent otitis media with effusion (OME). We therefore investigated the expression of PRRs and immunoglobulins in children with OME. The study population consisted of 66 children with OME, of whom 27 had more than 4 episodes in 12 months or more than 3 episodes in 6 months (otitis-prone group), and 39 had fewer than 4 episodes in 12 months or 3 episodes in 6 months (non-otitis-prone group). The expression in middle ear effusion of Toll-like receptor (TLR)-9, nucleotide-binding oligomerization domain (NOD)-1, NOD-2, and retinoic acid-inducible gene (RIG)-I mRNA, as determined by real-time PCR, and the concentrations of IgG, IgA, and IgM, as determined by ELISA, were compared between the two groups. The levels of TLR-9, NOD-1 and RIG I mRNAs were significantly lower in the otitis-prone than in the non-otitis-prone group (p<0.05 each). The concentrations of IgG, IgA and IgM in effusion fluid did not differ significantly between the two groups (p>0.05), and there were no correlations between immunoglobulin concentration and the expression of PRPs (p>0.05). Decreased expression of PRRs may be associated with increased susceptibility to OME.


Targeting the correct HDAC(s) to treat cognitive disorders. Changes in gene expression in the brain may underlie cognitive deficits inherent to normal aging and neurodegenerative disease. However, the mechanisms underlying pathological alterations in the brain transcriptome are incompletely understood. Epigenetic mechanisms such as DNA methylation and histone acetylation have been shown to be important for memory processes in the adult brain. There is accumulating evidence that altered chromatin plasticity and histone acetylation are also involved in cognitive aging, neurodegeneration, and neuropsychiatric diseases. Inhibitors of histone deacetylase (HDAC) exhibit neuroprotective and neuroregenerative properties in animal models of various brain diseases. As such, targeting of HDACs seems to be a promising therapeutic strategy. In this review, we discuss the specific roles of each HDAC protein and the possible function of distinct histone modifications. We hope that this knowledge will aid in the development of diagnostic tools and in designing more potent and specific treatment for neurological disorders targeting selective HDAC proteins.


Is adult neurogenesis essential for olfaction? In mammals, new neurons are recruited into restricted brain areas throughout life. Adult-born neurons produced in the subventricular zone of the lateral ventricle migrate rostrally towards the olfactory bulb. Although thousands of neurons reach this central structure every day, the functional impact of their integration into mature circuits remains a matter of debate. Recent investigations have revealed no striking sensory deficits per se when adult bulbar neurogenesis is challenged. However, some cognitive functions, such as perceptual learning and olfactory memory, are clearly impaired. In this review we highlight the role of network activity in shaping ongoing neurogenesis and, in turn, how the integration of adult-born neurons refines pre-existing network function, and consequently olfactory behavior.


Evaluating the Type II error rate in a sediment toxicity classification using the Reference Condition Approach. Sediments from 71 river sites in Northern Spain were tested using the oligochaete Tubifex tubifex (Annelida, Clitellata) chronic bioassay. 47 sediments were identified as reference primarily from macroinvertebrate community characteristics. The data for the toxicological endpoints were examined using non-metric MDS. Probability ellipses were constructed around the reference sites in multidimensional space to establish a classification for assessing test-sediments into one of three categories (Non Toxic, Potentially Toxic, and Toxic). The construction of such probability ellipses sets the Type I error rate. However, we also wished to include in the decision process for identifying pass-fail boundaries the degree of disturbance required to be detected, and the likelihood of being wrong in detecting that disturbance (i.e. the Type II error). Setting the ellipse size to use based on Type I error does not include any consideration of the probability of Type II error. To do this, the toxicological response observed in the reference sediments was manipulated by simulating different degrees of disturbance (simpacted sediments), and measuring the Type II error rate for each set of the simpacted sediments. From this procedure, the frequency at each probability ellipse of identifying impairment using sediments with known level of disturbance is quantified. Thirteen levels of disturbance and seven probability ellipses were tested. Based on the results the decision boundary for Non Toxic and Potentially Toxic was set at the 80% probability ellipse, and the boundary for Potentially Toxic and Toxic at the 95% probability ellipse. Using this approach, 9 test sediments were classified as Toxic, 2 as Potentially Toxic, and 13 as Non Toxic.


Assessing combined toxicity of estrogen receptor agonists in a primary culture of rainbow trout (Oncorhynchus mykiss) hepatocytes. The presence of highly complex mixtures of chemicals in the environment challenges our ability to assess single chemical effects and the interaction that occurs with cellular receptor targets and regulation of endocrine processes. In this study concentration addition (CA) and independent action (IA) prediction models were used to assess the combined toxicity of mixtures of environmental relevant estrogen receptor (ER) agonists (hormones and anthropogenic pollutants) in a primary culture of rainbow trout (Oncorhynchus mykiss) hepatocytes using the ER-mediated production of vitellogenin (Vtg) as a biological marker (biomarker) for estrogenicity. Nine of the eleven tested chemicals induced the production of Vtg and the parameters from the fitted concentration-response curves were used to model four mixtures containing four (17β-estradiol, estrone, estriol and diethylstilbestrol), five (musk ketone, 4-tert-octylphenol, bisphenol A, o,p'-DDT and dibenzothiophene), seven (17β-estradiol, estrone, estriol, diethylstilbestrol, 4-tert-octylphenol, bisphenol A and o,p'-DDT) and nine compounds (17β-estradiol, estrone, estriol, diethylstilbestrol, musk ketone, 4-tert-octylphenol, bisphenol A, o,p'-DDT and dibenzothiophene). The CA and IA prediction model proved to be a good estimation for the combined effect of mixtures of ER agonists at low relative mixture concentration (e.g. relative to the maximum mixture concentrations used), but a deviation from the prediction models was observed when exposing hepatocytes to high relative mixture concentrations. The CA and IA prediction models' ability to predict the combined estrogenic effect of complex mixtures, especially in the low concentration-response range, is of ecological relevance since organisms in the environment generally encounter low concentrations of chemicals from a wide array of chemical groups that may not elicit estrogenic effects on their own.


Fathead minnow (Pimephales promelas) reproduction is impaired in aged oil sands process-affected waters. Large volumes of fluid tailings are generated during the extraction of bitumen from oil sands. As part of their reclamation plan, oil sands operators in Alberta propose to transfer these fluid tailings to end pit lakes and, over time, these are expected to develop lake habitats with productive capabilities comparable to natural lakes in the region. This study evaluates the potential impact of various oil sands process-affected waters (OSPW) on the reproduction of adult fathead minnow (Pimephales promelas) under laboratory conditions. Two separate assays with aged OPSW (>15 years) from the experimental ponds at Syncrude Canada Ltd. showed that water containing high concentrations of naphthenic acids (NAs; >25 mg/l) and elevated conductivity (>2000 μS/cm) completely inhibited spawning of fathead minnows and reduced male secondary sexual characteristics. Measurement of plasma sex steroid levels showed that male fathead minnows had lower concentrations of testosterone and 11-ketotestosterone whereas females had lower concentrations of 17β-estradiol. In a third assay, fathead minnows were first acclimated to the higher salinity conditions typical of OSPW for several weeks and then exposed to aged OSPW from Suncor Energy Inc. (NAs ∼40 mg/l and conductivity ∼2000 μS/cm). Spawning was significantly reduced in fathead minnows held in this effluent and male fathead minnows had lower concentrations of testosterone and 11-ketotestosterone. Collectively, these studies demonstrate that aged OSPW has the potential to negatively affect the reproductive physiology of fathead minnows and suggest that aquatic habitats with high NAs concentrations (>25 mg/l) and conductivities (>2000 μS/cm) would not be conducive for successful fish reproduction.


Spheric balloon technique for ostial bifurcation lesion treatment.


Peripheral chemosensor function is blunted in moderate to severe chronic kidney disease. Cardiovascular mortality is markedly increased in chronic kidney disease (CKD) and may be explained in part by sympathetic hyperactivity. Impaired hyperoxic chemoreflex sensitivity (CHRS) has been attributed to an increased sympathetic tone. The aim of the present study was to examine whether chemosensor function is altered in patients with CKD. We assessed CHRS in 20 patients with stage 3 CKD [glomerular filtration rate (GFR) 30-59 ml/min/1.73 m(2)], in 15 patients with stage 4 CKD [GFR 15-29 ml/min/1.73 m(2)], as well as in 35 age and gender matched patients without any evidence of CKD. The difference in the R-R intervals divided by the difference in the oxygen pressures before and after deactivation of the chemoreceptors by inhalation of pure oxygen was calculated as the CHRS. A CHRS below 3.0 ms/mmHg was defined as pathological. CHRS was significantly depressed in patients with stage 3 CKD (2.9 ± 0.9 ms/mmHg, P=0.005) and in patients with stage 4 CKD (2.1 ± 0.6 ms/mmHg, P<0.001), as compared with patients without CKD (6.7 ± 0.9 ms/mmHg). There was a negative correlation between serum creatinine and CHRS (r=-0.51; P<0.001). In patients with CKD, chemosensor deactivation decreased mean arterial pressure from 91 ± 4 mmHg to 87 ± 3 mmHg (P=0.03). Multivariate analysis showed that GFR (P=0.001) was the only independent predictor of a pathological CHRS. Using a relatively non-invasive bedside test we provide evidence for a blunted peripheral chemosensor function in chronic kidney disease. We thereby lay the basis for interventional studies assessing chemosensor function in chronic kidney disease.


Field trial of a synthetic tsetse-repellent technology developed for the control of bovine trypanosomosis in Kenya. We conducted a field trial among Maasai cattle-keepers in Nkuruman and Nkineji areas of Kenya to evaluate the effectiveness of a synthetic tsetse-repellent technology developed for the control of trypanosomosis in cattle. The technology was a repellent (2-methoxy 4-methylphenol) emitted from dispensers attached to collars worn by cattle. Treatment was allocated at the herd level to ensure adequate protection of all the animals in a herd, with measurements of effectiveness conducted at the individual-animal level. The trial began in April 2005 and ran for 16 months including a baseline phase of 4 months. We recruited 12 herds in each area using a restricted random-sampling technique and distributed them equally into intervention (repellent) and control groups. Sample size was determined using a formal power calculation. Effectiveness or minimal worthwhile difference was defined as a 50% reduction in the incidence of trypanosome infection in the treated versus control group (effectiveness below which the technology was considered by experts as not viable compared to existing control techniques). All the animals in the recruited herds were screened monthly (buffy-coat technique) for trypanosome infections. The analysis followed the principle of intention-to-treat by which subjects are analysed according to their initial treatment assignment, regardless of the mechanical performance of the device. Crude and adjusted effects of the technology were 23% (p<0.001) and 18% (p=0.08) reduction in the infection incidence in the treatment compared to the control groups, respectively. The impact of the technology estimated in this study did not achieve the threshold of 50% reduction in the trypanosome infection incidence set a priori to indicate effectiveness (p<0.001). We therefore concluded that the prototype repellent technology package was not sufficiently effective in reducing trypanosome infection incidence under natural tsetse challenge to merit commercial development.


Geochemical interactions between process-affected water from oil sands tailings ponds and North Alberta surficial sediments. In Northern Alberta, the placement of out-of-pit oil sands tailings ponds atop natural buried sand channels is becoming increasingly common. Preliminary modeling of such a site suggests that process-affected (PA) pond water will infiltrate through the underlying clay till aquitard, reaching the sand channel. However, the impact of seepage upon native sediments and groundwater resources is not known. The goal of this study is to investigate the role of adsorption and ion exchange reactions in the clay till and their effect on the attenuation or release of inorganic species. This was evaluated using batch sorption experiments (traditional and a recent modification using less disturbed sediment samples) and geochemical modeling with PHREEQC. The results show that clay till sediments have the capacity to mitigate the high concentrations of ingressing sodium (600 mg L(-1)), with linear sorption partitioning coefficients (K(d)) of 0.45 L kg(-1). Ion exchange theory was required to account for all other cation behaviour, precluding the calculation of such coefficients for other species. Qualitative evidence suggests that chloride will behave conservatively, with high concentrations remaining in solution (375 mg L(-1)). As a whole, system behaviour was found to be controlled by a combination of competitive ion exchange, dissolution and precipitation reactions. Observations, supported by PHREEQC simulations, suggest that the influx of PA water will induce the dissolution of pre-existing sulphate salts. Sodium present in the process-affected water will exchange with sediment-bound calcium and magnesium, increasing the divalent ions' pore fluid concentrations, and leading to the precipitation of a calcium-magnesium carbonate mineral phase. Thus, in similar tailings pond settings, particularly if the glacial till coverage is thin or altogether absent, it is reasonable to expect that high concentrations of sodium and chloride will remain in solution, while sulphate concentrations will exceed those of the ingressing plume (150 mg L(-1)).


Proteome analysis of sugar beet (Beta vulgaris L.) elucidates constitutive adaptation during the first phase of salt stress. Salinity is one of the major stress factors responsible for growth reduction of most of the higher plants. In this study, the effect of salt stress on protein pattern in shoots and roots of sugar beet (Beta vulgaris L.) was examined. Sugar beet plants were grown in hydroponics under control and 125 mM salt treatments. A significant growth reduction of shoots and roots was observed. The changes in protein expression, caused by salinity, were monitored using two-dimensional gel-electrophoresis. Most of the detected proteins in sugar beet showed stability under salt stress. The statistical analysis of detected proteins showed that the expression of only six proteins from shoots and three proteins from roots were significantly altered. At this stage, the significantly changed protein expressions we detected could not be attributed to sugar beet adaptation under salt stress. However, unchanged membrane bound proteins under salt stress did reveal the constitutive adaptation of sugar beet to salt stress at the plasma membrane level.


Overexpression of CB2 cannabinoid receptors results in neuroprotection against behavioral and neurochemical alterations induced by intracaudate administration of 6-hydroxydopamine. The role of CB2 cannabinoid receptors in the behavioral and neurochemical changes induced by intracaudate administration of 6-hydroxydopamine (6-OHDA) was evaluated. 6-OHDA (12 μg/4 μL) or its vehicle was injected in the caudate-putamen (CPu) of mice overexpressing the CB2 cannabinoid receptor (CB2xP) and wild type (WT) mice. Motor impairment, emotional behavior, and cognitive alterations were evaluated. Tyrosine hydroxylase (TH), glial fibrillary acidic protein (GFAP), and ionized calcium-binding adapter molecule 1 (Iba-1) were measured by immunocytochemistry in the CPu and/or substantia nigra (SN) of CB2xP mice and WT mice. Oxidative/nitrosative and neuroinflammatory parameters were also measured in the CPu and cortex of 6-OHDA-treated and sham-treated mice. 6-OHDA-treated CB2xP mice presented significantly less motor deterioration than 6-OHDA-treated WT mice. Immunocytochemical analysis of tyrosine hydroxylase in the SN and CPu revealed significantly fewer lesions in CB2xP mice than in WT mice. GFAP and Iba-1 immunostaining revealed less astrocyte and microglia recruitment to the treated area of the CPu in CB2xP mice. Malonyldialdehyde (MDA) concentrations were lower in the striatum and cerebral cortex of sham-treated CB2xP mice than in sham-treated WT mice. The administration of 6-OHDA increased MDA levels in both WT mice and CB2xP mice; it increased the oxidized (GSSG)/reduced (GSH) glutathione ratio in the striatum in WT mice alone compared with matched sham-treated controls. The results revealed that overexpression of CB2 cannabinoid receptors decreased the extent of motor impairment and dopaminergic neuronal loss, reduced the recruitment of astrocytes and microglia to the lesion, and decreased the level of various oxidative parameters. These results suggest that CB2 receptors offer neuroprotection against dopaminergic injury.


SAP97-mediated local trafficking is altered in Alzheimer disease patients' hippocampus. Synapse-asssociated protein-97 (SAP97) is responsible for the trafficking of both glutamate receptor subunits, GluR1 and NR2A, and α-secretase ADAM10 to the synaptic membrane. Here we evaluate the trafficking capability of SAP97 in Alzheimer disease (AD) patients' brain. We analyzed autoptic hippocampus and superior frontal gyrus, respectively as an affected and a less affected area, from 6 AD patients (Braak 4) and 6 healthy controls. In hippocampus, but not in superior frontal gyrus, of AD patients, ADAM10 and GluR1 synaptic membrane levels are altered while NR2A localization is not affected. Both immunoprecipitation and pull-down assays demonstrated that SAP97 failed to correctly couple to ADAM10 and GluR1, but not to NR2A. These findings not only indicate SAP97 as a point of convergence between amyloid cascade and synaptic failure in AD, but also allow a different interpretation of AD which can be now perceived as synaptic trafficking defect pathology.


Serum fatty acid profiles using GC-MS and multivariate statistical analysis: potential biomarkers of Alzheimer's disease. Previous studies showed the relationship between fatty acids and the risk of developing Alzheimer's disease (AD). However, they did not address potential differences in free fatty acid (FFA) profiles that could be used to distinguish between AD patients and healthy controls. In the present study we used gas chromatography-mass spectrometry (GC-MS) technology coupled with multivariate statistical analysis to study profiles of FFA in AD. The results indicated 2 saturated fatty acids (C14:0 and C16:0; p < 0.001 and p < 0.05, respectively), 3 unsaturated fatty acids (C18:1, C18:3, and C22:6; p < 0.05, p < 0.05, and p < 0.001, respectively), where mean levels in serum from AD patients were significantly lower than controls. Partial least squares discriminant analysis (PLS-DA) models with unit variance (UV) scaling and orthogonal signal correction (OSC) data preprocessing methods were employed to refine intergroup differences between FFA profiles. The results of the analysis have highlighted docosahexaenoic acid (DHA) as the FFA with the greatest potential as a biomarker of AD, and this study has demonstrated that FFA biomarkers have considerable potential in diagnosing and monitoring AD.


A neuroprotective sulfone of marine origin and the in vivo anti-inflammatory activity of an analogue. Our continuing effort of searching bioactive substances from the Formosan soft coral Cladiella australis has led to the isolation of a bioactive substance austrasulfone (1), which possesses significant neuroprotective activities. A straightforward synthesis of 1 was achieved by a two-step reaction sequence. Dihydroaustrasulfone alcohol (3), the synthetic precursor of 1, not only exhibited in vitro anti-inflammatory activity, but also showed potent therapeutic ability in the treatment of neuropathic pain, atherosclerosis, and multiple sclerosis in rats.


The impact of informal caregivers on depressive symptoms among older adults receiving formal home health care. This study evaluated the association between presence and types of informal caregivers and the presence of depressive symptoms among older adults receiving formal home health care (HHC). A secondary analysis of data was conducted using a computerized patient care database, the Outcome and Assessment Information Set. Logistic regression analyses were used to examine the data of 8448 patients aged 65 years or older who had been admitted to an HHC agency from acute care hospitals between January 1, 2002 and June 30, 2002. The outcome variable was the presence of depressive symptoms. The primary predictor variable was the presence and types of informal caregivers. Covariates included demographic variables, health status, length of time enrolled in formal HHC, patient living arrangements, and the frequency and types of care received from informal caregivers. A lower percentage of older adults receiving care from both informal caregivers and a formal HHC agency (13.3%) had depressive symptoms than older adults receiving only formal HHC (14.9%) at the end of a 60-day episode in formal HHC. Older adults without an informal caregiver were more likely to experience depressive symptoms than those with an informal caregiver after a 60-day episode in HHC (odds ratio = 1.229, 95% confidence interval = 1.027-1.471). There was no significant association between the types of informal caregivers and the presence of depressive symptoms.


Biological evaluation of donor-acceptor aminonaphthoquinones as antitumor agents. Several members of the phenylamino-1,4-naphthoquinone series were prepared in order to investigate structure-activity relationships (SAR) and to explore the antitumor effects associated with this scaffold. The cytotoxic effects of the aminoquinones (EC50) against a panel of cancer cell lines (MCF7, DU145 and T24 cells) and healthy fibroblasts (BALB/3T3) were assessed in vitro using the MTT reduction assay 48 h after drug exposure. SAR analysis of the aminonaphthoquinone series showed that insertion of a chlorine atom in the acceptor quinone nucleus and/or insertion of a methyl group at the nitrogen atom of the donor phenylamino group induced significant changes in cytotoxic activity. Quinones 7 and 9, which exhibited the highest selective indexes (5.73 and 6.29, respectively), were further characterized using the following assays: Colony formation, caspase-3 activity, and ATP content. The results showed that aminoquinone 7 strongly influenced ATP levels and impaired the proliferative capacity of T24 cells without activating caspase-3.


Serum paraoxonase activity is associated with variants in the PON gene cluster and risk of Alzheimer disease. Previous studies have shown association of single nucleotide polymorphisms (SNPs) in 3 contiguous genes (PON1, PON2, and PON3) encoding paraoxonase with risk of Alzheimer disease (AD). We evaluated the association of serum paraoxonase activity measured by phenyl acetate (PA) and thiobutyl butyrolactone (TBBL) with risk of AD and with 26 SNPs spanning the PON gene cluster in 266 AD cases and 306 sibling controls from the MIRAGE study. The odds of AD (adjusted for age, gender, and ethnicity) increased 20% for each standard deviation decrease in PA or TBBL activity. There were association signals with activity in all 3 genes. Haplotypes including SNPs spanning the PON genes were generally more significant than haplotypes comprising SNPs from 1 gene. Significant interactions were observed between SNP pairs located across the PON cluster with either serum activity measure as the outcome, and between several PON SNPs and PA activity with AD status as the outcome. Our results suggest that low serum paraoxonase activity is a risk factor for AD. Furthermore, multiple variants in PON influence serum paraoxonase activity and their effects may be synergistic.


Forest fires in the former Soviet Union: no reasons for radiophobia.


Development of an energy discriminate CR-39(®) nuclear track etch dosimeter for Radon-220 gas measurements. An energy discriminate CR-39(®) nuclear track etch dosimeter for use in a (220)Rn and (222)Rn gas monitor has been developed and experimentally assessed. It utilises a thin film of Mylar(®) C to attenuate the alpha particle energies to allow only the damage tracks created by the 8.785 MeV alpha particles emitted from (212)Po of the (232)Th decay chain to be registered in the CR-39(®) plaque, allowing for the direct measurement of (220)Rn gas concentrations. The dosimeter was developed through a combination of experimental investigations and theoretical simulations using the Monte Carlo ion transport modelling program Stopping and Range of Ions in Materials (SRIM 2008). A film thickness of 54 μm has been shown to attenuate all alpha energies less then 7.7 MeV.


Re.: Development of in-house rapid manufacturing of three-dimensional models in maxillofacial surgery.


Environmental quality of mussel farms in the Vigo estuary: pollution by PAHs, origin and effects on reproduction. This work analyzes the influence of environmental and physiological parameters on PAHs accumulation in cultured mussels. Lipid content and reproductive stage are directly related with PAHs accumulation pattern. We observed a rapid accumulation and depuration of PAHs, mainly during periods of nutrients accumulation, spawns and gonadic restorations. Correlations between PAHs accumulation and physiological status indicate when mussels are more susceptible to adverse effects of these pollutants. A positive correlation between mutagenic congener's accumulation and occurrence of gonadic neoplastic disorders is shown for the first time in mussels. Molecular indices were used to identify the origin of hydrocarbons accumulated by Mytilus, showing a chronic pyrolytic pollution and pollutant episodes by petrogenic sources and biomass combustion in the studied area. Multivariate analysis suggests the possibility of including physiological parameters of sentinel organisms in environmental biomonitoring programs, mainly in aquaculture areas, taking into account their two aspects: farms productivity and human food safety.


The physiological, transcriptional and genetic responses of an ozone-sensitive and an ozone tolerant poplar and selected extremes of their F2 progeny. Relatively little is known about the transcriptional response or genetic control of response and adaptation of trees to tropospheric ozone exposure. Such understanding is needed as up to 50% of forests, globally, may be subjected to phytotoxic concentrations of ozone. The physiological, transcriptional and genetic response to ozone was examined in Populus trichocarpa and P. deltoides, which show extreme sensitivity and tolerance to ozone, respectively. Using an inbred F2 mapping population derived from these two species, we mapped quantitative trait loci (QTL) for traits associated with ozone response, examined segregation of the transcriptional response to ozone and co-located genes showing divergent responses between tolerant and sensitive genotypes with QTL. QTL were identified linking detrimental effects of ozone with leaf and biomass traits and differential responses were found for key genes involved in ethylene production and response.


Neighborhoods, daily activities, and measuring health risks experienced in urban environments. Studies of place and health often classify a subject's exposure status according to that which is present in their neighborhood of residence. One's neighborhood is often proxied by designating it to be an administratively defined unit such as census tract, to make analysis feasible. Although it is understood that residential space and actual lived space may not correspond and therefore exposure misclassification may result, few studies have the opportunity to investigate the implications of this issue concretely. A population-based case-control study that is currently underway provides one such opportunity. Adolescent victims of assault in Philadelphia, Pennsylvania, USA, and a control sample of adolescents drawn randomly from the community are being enrolled to study how alcohol consumption and time spent nearby alcohol outlets - individual-level and environmental-level risk factors for violence, respectively - over the course of daily activities relate to the likelihood of being assaulted. Data from a rapport-building exercise consist of hand-drawn sketches that subjects drew on street maps when asked to indicate the area considered their neighborhood. The main data consist of self-reported, detailed paths of the routes adolescents traveled from one location to the next over the course of one full day. Having noticed interesting patterns as the data collection phase proceeds, we present here an analysis conducted with the data of 55 control subjects between 15 and 19 years old. We found that hand-drawn neighborhoods and activity paths did not correspond to census tract boundaries, and time subjects spent in close proximity to alcohol outlets during their daily activities was not correlated with the prevalence of alcohol outlets in the census tract of their residence. This served as a useful example demonstrating how classifying subjects as exposed based solely on the prevalence of the exposure in the geographic area of their residence may misrepresent the exposure that is etiologically meaningful.


Infant mortality, season of birth and the health of older Puerto Rican adults. The increasing prevalence of heart disease and diabetes among aging populations in low and middle income countries leads to questions regarding the degree to which endogenous early life exposures (exposures in utero) are important determinants of these health conditions. We devised a test using infant mortality (IMR) to verify if season of birth is a good indicator of early life (in utero) conditions that precipitate adult onset of disease. We linked annual IMR at the municipality (municipio) level from the late 1920s to early 1940s with individual birth year and place using a representative sample of older Puerto Rican adults (n = 1447) from the Puerto Rican Elderly: Health Conditions (PREHCO) study. We estimated the effects of season of birth on adult heart disease and diabetes for all respondents and then for respondents according to whether they were born when IMR was lower or higher, controlling for age, gender, obesity, respondent's educational level, adult behavior (smoking and exercise) and other early life exposures (childhood health, knee height and childhood socioeconomic status (SES)). The pattern of effects suggests that season of birth reflects endogenous causes: (1) odds of heart disease and diabetes were strong and significant for those born during the lean season in years when IMR was lower; (2) effects remained consistent even after controlling for other childhood conditions and adult behavior; but (3) no seasonality effects on adult health for adults born when IMR was higher. We conclude that in this population of older Puerto Rican adults there is continued support that the timing of adverse endogenous (in utero) conditions such as poor nutrition and infectious diseases is associated with adult heart disease and diabetes. It will be important to test the validity of these findings in other similar populations in the developing world.


National female literacy, individual socio-economic status, and maternal health care use in sub-Saharan Africa. The United Nations Millennium Development Goals have identified improving women's access to maternal health care as a key target in reducing maternal mortality in sub-Saharan Africa (sSA). Although individual factors such as income and urban residence can affect maternal health care use, little is known about national-level factors associated with use. Yet, such knowledge may highlight the importance of global and national policies in improving use. This study examines the importance of national female literacy on women's maternal health care use in continental sSA. Data that come from the 2002-2003 World Health Survey. Multilevel logistic regression was used to examine the association between national female literacy and individual's non-use of maternal health care, while adjusting for individual-level factors and national economic development. Analyses also assessed effect modification of the association between income and non-use by female literacy. Effect modification was evaluated with the likelihood ratio test (G(2)). We found that within countries, individual age, education, urban residence and household income were associated with lack of maternal health care. National female literacy modified the association of household income with lack of maternal health care use. The strength of the association between income and lack of maternal health care was weaker in countries with higher female literacy. We conclude therefore that higher national levels of female literacy may reduce income-related inequalities in use through a range of possible mechanisms, including women's increased labour participation and higher status in society. National policies that are able to address female literacy and women's status in sub-Saharan Africa may help reduce income-related inequalities in maternal health care use.


Hierarchy of local structural and dynamics perturbations due to subdenaturing urea in the native state ensemble of DLC8 dimer. Local structural and dynamic modulations due to small environmental perturbations reflect the adaptability of the protein to different interactors. We have investigated here the preferential local perturbations in Dynein light chain protein (DLC8), a cargo adapter, by sub-denaturing urea concentrations. Equilibrium unfolding experiments by optical spectroscopic methods indicated a two state like unfolding of DLC8 dimer, with the transition mid-point occurring around 8.6M urea. NMR studies identified the β3 and β4 strands, N-, C- terminal regions, loops connecting β1 to α1, α1 to α2 and β3 to β4 as the soft targets of urea perturbation and thus indicated potential unfolding initiation sites. Native-state hydrogen exchange studies suggested the unfolding to traverse from the edges towards the centre of the secondary structural elements. At 6M urea the whole protein chain acts like a cooperative unit. These observations are expected to have important implications for the protein's multiple functions.


Inhibition of mir-21, which is up-regulated during MYCN knockdown-mediated differentiation, does not prevent differentiation of neuroblastoma cells. Neuroblastoma is a malignant childhood tumour arising from precursor cells of the sympathetic nervous system. Genomic amplification of the MYCN oncogene is associated with dismal prognosis. For this group of high-risk tumours, the induction of tumour cell differentiation is part of current treatment protocols. MicroRNAs (miRNAs) are small non-coding RNA molecules that effectively reduce the translation of target mRNAs. MiRNAs play an important role in cell proliferation, apoptosis, differentiation and cancer. In this study, we investigated the role of N-myc on miRNA expression in MYCN-amplified neuroblastoma. We performed a miRNA profiling study on SK-N-BE (2) cells, and determined differentially expressed miRNAs during differentiation initiated by MYCN knockdown, using anti-MYCN short-hairpin RNA (shRNA) technology. Microarray analyses revealed 23 miRNAs differentially expressed during the MYCN knockdown-mediated neuronal differentiation of MNA neuroblastoma cells. The expression changes were bidirectional, with 11 and 12 miRNAs being up- and down-regulated, respectively. Among the down-regulated miRNAs, we found several members of the mir-17 family of miRNAs. Mir-21, an established oncomir in a variety of cancer types, became strongly up-regulated upon MYCN knockdown and the subsequent differentiation. Neither overexpression of mir-21 in the high-MYCN neuroblastoma cells, nor repression of increased mir-21 levels during MYCN knockdown-mediated differentiation had any significant effects on cell differentiation or proliferation. We describe a subset of miRNAs that were altered during the N-myc deprived differentiation of MYCN-amplified neuroblastoma cells. In this context, N-myc acts as both an activator and suppressor of miRNA expression. Mir-21 was up-regulated during cell differentiation, but inhibition of mir-21 did not prevent this process. We were unable to establish a role for this miRNA during differentiation and proliferation of the two neuroblastoma cell lines used in this study.


Evaluating the PM damage cost due to urban air pollution and vehicle emissions in Seoul, Korea. This study evaluated the prospective damage costs of PM(2.5) inhalation. We performed a health risk assessment based on an exposure-response function to estimate the annual population risk in the Seoul metropolitan city, Korea. Also, we estimated a willingness-to-pay (WTP) amount for reducing the mortality rate in order to evaluate a statistical life value. We combined the annual population risk and the value-of-statistical-life to calculate the damage cost estimate. In the health risk assessment, we applied the PM(2.5) relative risk to evaluate the annual population risk. We targeted an exposure population of 5,401,369 persons who were over the age of 30. Using a Monte-Carlo simulation for uncertainty analysis, we estimated that the population risk of PM(2.5) inhalation during a year in Seoul is 2181 premature deaths for acute exposure and 18,510 premature deaths for chronic exposure. The monthly average WTP for 5/1000 mortality reduction over ten years is $20.20 USD (95% C.I: $16.60-24.50) and the implied value-of-statistical-life (VSL) is $485,000 USD (95% C.I: $398,000-588,000). The damage cost estimate due to risk from PM(2.5) inhalation in Seoul is about $1057 million USD per year for acute exposure, and $8972 million USD per year for chronic exposure. It is important to note that this cost estimate does not reflect all health damage cost estimates in this urban area. This recommendation is a model for evaluating a mortality risk reduction and as such we must re-evaluate an integrated application of morbidity risk.


Sorption of hydrophobic pesticides on a Mediterranean soil affected by wastewater, dissolved organic matter and salts. Irrigation with treated wastewaters as an alternative in countries with severe water shortage may influence the sorption of pesticides and their environmental effects, as wastewater contains higher concentrations of suspended and dissolved organic matter and inorganic compounds than freshwater. We have examined the sorption behaviour of three highly hydrophobic pesticides (the herbicide pendimethalin and the insecticides α-cypermethrin and deltamethrin) on a Mediterranean agricultural soil using the batch equilibration method. We considered wastewater, extracts from urban sewage sludge with different dissolved organic carbon contents, and inorganic salt solutions, using Milli Q water as a control. All pesticides were strongly retained by soil although some sorption occurred on the walls of the laboratory containers, especially when wastewater and inorganic salt solutions were used. The calculation of distribution constants by measuring pesticide concentrations in soil and solution indicated that pendimethalin sorption was not affected whereas α-cypermethrin and deltamethrin retention were significantly enhanced (ca. 5 and 2 times, respectively) when wastewater or salt solutions were employed. We therefore conclude that the increased sorption of the two pesticides caused by wastewater cannot be only the result of its dissolved organic carbon content, but also of the simultaneous presence of inorganic salts in the solution.


Index models to evaluate the risk of phosphorus and nitrogen loss at catchment scales. This paper investigates index models as a tool to estimate the risk of N and P source strengths and loss at the catchment scale. The index models assist managers in improving the focus of remediation actions that reduce nutrient delivery to waterbodies. N and P source risk factors (e.g. soil nutrient concentrations) and transport risk factors (e.g. distance-to-streams) are used to determine the overall risk of nutrient loss for a case study in the Tuross River catchment of coastal southeast Australia. In the development of the N index model for Tuross, particulate N was considered important based on the observed event water quality data. In contrast to previous N index models, erosion and contributing distance were therefore included in the Tuross River catchment N index. Event-based water quality monitoring, and soil information, or in data-poor catchments conceptual understanding, are essential to represent catchment-scale processes. The techniques have high applicability in other catchments, and are complementary to other modelling techniques such as process-based semi-distributed modelling. Index models generally provide much more detailed spatial resolution than fully- or semi-distributed conceptual modelling approaches. Semi-distributed models can be used to quantify nutrient loads and provide overall direction to set the broad focus of management. Index models can then be used to refine on-the-ground investigations and investment priorities. In this way semi-distributed models can be combined with index models to provide a set of powerful tools to influence management decisions and outcomes.


Re: Ricardo L. Favaretto, Shahrokh F. Shariat, Daher C. Chade, et al. The effect of tumor location on prognosis in patients treated with radical nephroureterectomy at Memorial Sloan-Kettering Cancer Center. Eur Urol 2010;58:574-580.


Ricardo L. Favaretto, Shahrokh F. Shariat, Daher C. Chade, et al. Comparison between laparoscopic and open radical nephroureterectomy in a contemporary group of patients: are recurrence and disease-specific survival associated with surgical technique? Eur Urol 2010;58:645-651.


Prognostic impact of the 2009 UICC/AJCC TNM staging system for renal cell carcinoma with venous extension. The prognostic significance of venous involvement and tumour thrombus level in renal cell carcinoma (RCC) remains highly controversial. In 2010, the American Joint Committee on Cancer (AJCC) and the Union International Centre le Cancer (UICC) revised the RCC staging system (7th edition) based on tumour thrombus level, differentiating the T stage of tumours limited to renal-vein-only involvement. We aimed to evaluate the impact of tumour thrombus extension in a multi-institutional cohort of patients. An international consortium of 11 institutions was established to retrospectively review a combined cohort of 1215 patients undergoing radical nephrectomy and tumour thrombectomy for RCC, including 585 patients with inferior vena cava (IVC) involvement or higher. Predictive factors of survival, including histology, tumour thrombus level, nodal status, Fuhrman grade, and tumour size, were analysed. A total of 1122 patients with complete data were reviewed. The median follow-up for all patients was 24.7 mo, with a median survival of 33.8 mo. The 5-yr survival was 43.2% (renal vein involvement), 37% (IVC below the diaphragm), and 22% with caval involvement above the diaphragm. On multivariate analysis, tumour size (hazard ratio [HR]: 1.64 [range: 1.03-2.59]; p=0.036), Fuhrman grade (HR: 2.26 [range: 1.65-3.1]; p=0.000), nodal metastasis (HR: 1.32 [range: 1.09-1.67]; p=0.005), and tumour thrombus level (HR: 2.10 [range: 1.53-3.0]; p=0.00) correlated independently with survival. Based on analysis of the largest known cohort of patients with RCC along with IVC and atrial thrombus involvement, tumour thrombus level is an independent predictor of survival. Our findings support the changes to the latest AJCC/UICC staging system.


Critical assessment of preoperative urinary prostate cancer antigen 3 on the accuracy of prostate cancer staging. Knowledge about the staging significance of the prostate cancer antigen 3 (PCA3) score to better identify pathologic features after radical prostatectomy (RP) is limited and controversial. Our aim was to study the clinical staging significance of PCA3 to identify pathologic favorable and/or unfavorable features in the RP specimen. Complete retrospective clinical and pathologic data of consecutive men who had undergone RP from three tertiary referral centers including preoperative PCA3 scores (n=305) and computer-assisted planimetrically measured tumor volume data (n=160) were available. All patients were treated with RP. PCA3 scores were assessed using the PROGENSA assay (Gen-Probe, San Diego, CA, USA). Beyond standard risk factors (age, digital rectal examination, prostate-specific antigen, prostate volume, biopsy Gleason score, percentage of positive cores), five different PCA3 codings were used in logistic regression models to identify five distinct pathologic end points: (1) low-volume disease (<0.5 ml), (2) insignificant prostate cancer (PCa) according to the Epstein criteria, (3) extracapsular extension (ECE), (4) seminal vesicle invasion (SVI), and (5) aggressive disease defined as Gleason sum ≥7. Accuracy estimates of each end point were quantified using the area under the curve (AUC) of the receiver operator characteristic analysis in models with and without PCA3. PCA3 scores were significantly lower in low-volume disease and insignificant PCa (p ≤ 0.001). AUC of multivariable low-volume disease (+2.4 to +5.5%) and insignificant PCa models (+3 to +3.9%) increased when PCA3 was added to standard clinical risk factors. In contradistinction, regardless of its coding, PCA3 scores were not significantly elevated in pathologically confirmed ECE (p=0.4) or SVI (p=0.5), respectively. Higher PCA3 scores were associated with aggressive disease (p<0.001). Importantly, the addition of PCA3 to multivariable intermediate- and high-grade models did not improve prediction. Despite reporting the largest pathologic PCA3 study, the main limitation resides in its small sample size. PCA3 was confirmed as a valuable predictor of pathologically confirmed low-volume disease and insignificant PCa. Further exploration of its role as an additional marker to select patients for active surveillance may be warranted. In contradistinction, assessment of pathologically advanced or aggressive PCa is not improved using PCA3.


ZEB1-responsive genes in non-small cell lung cancer. The epithelial to mesenchymal transition (EMT) is a developmental process enabling epithelial cells to gain a migratory mesenchymal phenotype. In cancer, this process contributes to metastases; however the regulatory signals and mechanistic details are not fully elucidated. Here, we sought to identify the subset of genes regulated in lung cancer by ZEB1, an E-box transcriptional repressor known to induce EMT. Using an Affymetrix-based expression database of 38 non-small cell lung cancer (NSCLC) cell lines, we identified 324 genes that correlated negatively with ZEB1 and 142 that were positively correlated. A mesenchymal gene pattern (low E-cadherin, high Vimentin or N-cadherin) was significantly associated with ZEB1 and ZEB2, but not with Snail, Slug, Twist1 or Twist2. Among eight genes selected for validation, seven were confirmed to correlate with ZEB1 by quantitative real-time RT-PCR in a series of 22 NSCLC cell lines, either negatively (CDS1, EpCAM, ESRP1, ESRP2, ST14) or positively (FGFR1, Vimentin). In addition, over-expression or knockdown of ZEB1 led to corresponding changes in gene expression, demonstrating that these genes are also regulated by ZEB1, either directly or indirectly. Of note, the combined knockdown of ZEB1 and ZEB2 led to apparent synergistic responses in gene expression. Furthermore, these responses were not restricted to artificial settings, since most genes were similarly regulated during a physiologic induction of EMT by TGF-β plus EGF. Finally, the absence of ST14 (matriptase) was linked to ZEB1 positivity in lung cancer tissue microarrays, implying that the regulation observed in vitro applies to the human disease. In summary, this study identifies a new set of ZEB-regulated genes in human lung cancer cells and supports the hypothesis that ZEB1 and ZEB2 are key regulators of the EMT process in this disease.


Granular activated carbon promoted ozonation of a food-processing secondary effluent. This paper reports on the application of a simultaneous combination of ozone and a granular activated carbon (O(3)/GAC) as a tertiary treatment of a wastewater generated from the activity of various food-processing industries. Prior to the O(3)/GAC treatment, the wastewater was subjected to conventional primary and secondary treatments in a full-scale wastewater treatment plant (WWTP). The effluent from the WWTP presented high organic load (COD>500 mg/l and TOC>150 mg/l), which could be much reduced by the O(3)/GAC treatment. Results from the O(3)/GAC experiments were compared with those obtained in single ozonation, single adsorption onto GAC and sequential O(3)-GAC adsorption experiments. While single processes and the sequential one showed limited capacity to remove organic matter for the food-processing effluent (COD removal <40%), the simultaneous O(3)/GAC process led to decreases of COD up to 82% at the conditions here applied. The combined process also improved the ozone consumption, which decreased from about 19 g O(3)/g TOC (single ozonation process) to 8.2-10.7 g O(3)/g TOC (O(3)/GAC process). The reusability of the GAC throughout a series of consecutive O(3)/GAC experiments was studied with no apparent loss of activity for a neutral GAC (PZC = 6.7) but for a basic GAC (PZC = 9.1).


Carbon nanotube composite coated platinum electrode for detection of Ga(III). This study demonstrates the application of composite multi-walled carbon nanotube (MWCNT) polyvinylchloride (MWNT-PVC) based on 7-(2-hydroxy-5-methoxybenzyl)-5,6,7,8,9,10-hexahydro-2H benzo [b][1,4,7,10,13] dioxa triaza cyclopentadecine-3,11(4H,12H)-dione ionophore for gallium sensor. The sensor shows a good Nernstian slope of 19.68 ± 0.40 mV/decade in a wide linear range concentration of 7.9 × 10(-7) to 3.2 × 10(-2)M of Ga(NO(3))(3). The detection limit of this electrode is 5.2 × 10(-7)M of Ga(NO(3))(3). This proposed sensor is applicable in a pH range of 2.7-5.0. It has a short response time of about 10s and has a good selectivity over nineteen various metal ions. The practical analytical utility of this electrode is demonstrated by measurement of Ga(III) in river water.


Sad mood increases pain sensitivity upon thermal grill illusion stimulation: implications for central pain processing. In different fields of neuroscience research, illusions have successfully been used to unravel underlying mechanisms of stimulus processing. One such illusion existing for the field of pain research is the so-called thermal grill illusion. Here, painful sensations are elicited by interlacing warm and cold bars, with stimulus intensities (temperatures) of these bars being below the respective heat pain or cold pain thresholds. To date, the underlying mechanisms of this phenomenon are not completely understood. There is some agreement, however, that the sensation evoked by this stimulation is generated by central nervous interactions. Therefore, we followed two approaches in this study: firstly, we aimed at developing and validating a water-driven device which might be used in fMRI scanners in future studies - subject to minor adaptations. Secondly, we aimed to interfere with this illusion by induction of a sad mood state, a procedure which is suggested to influence central nervous structures that are also involved in pain processing. The newly developed device induced thermal grill sensations similar to those reported in the literature. Induction of sad, but not neutral mood states, resulted in higher pain and unpleasantness ratings of the painful illusion. These findings might be of importance for the understanding of pain processing in healthy volunteers, but putatively even more so in patients with major depressive disorder. Moreover, our results might indicate that central nervous structures involved in the affective domain or cognitive domain of pain processing might be involved in the perception of the illusion.


Veterinary parasitoses in the Mediterranean area.