Total Spots Research Articles

Genotoxic and Antigenotoxic Assessment of Chios Mastic Oil by the In Vitro Micronucleus Test on Human Lymphocytes and the In Vivo Wing Somatic Test on Drosophila.

Chios mastic oil (CMO), the essential oil derived from Pistacia lentiscus (L.) var. chia (Duham), has generated considerable interest because of its antimicrobial, anticancer, antioxidant and other beneficial properties. In the present study, the potential genotoxic activity of CMO as well as its antigenotoxic properties against the mutagenic agent mitomycin-C (MMC) were evaluated by employing the in vitro Cytokinesis Block MicroNucleus (CBMN) assay and the in vivo Somatic Mutation And Recombination Test (SMART). In the in vitro experiments, lymphocytes were treated with 0.01, 0.05 and 0.10% (v/v) of CMO with or without 0.05 μg/ml MMC, while in the in vivo assay Drosophila larvae were fed with 0.05, 0.10, 0.50 and 1.00% (v/v) of CMO with or without 2.50 μg/ml MMC. CMO did not significantly increase the frequency of micronuclei (MN) or total wing spots, indicating lack of mutagenic or recombinogenic activity. However, the in vitro analysis suggested cytotoxic activity of CMO. The simultaneous administration of MMC with CMO did not alter considerably the frequencies of MMC-induced MN and wing spots showing that CMO doesn’t exert antigenotoxic or antirecombinogenic action. Therefore, CMO could be considered as a safe product in terms of genotoxic potential. Even though it could not afford any protection against DNA damage, at least under our experimental conditions, its cytotoxic potential could be of interest.

Analysis and decomposition of active and reactive power spot price in deregulated electricity markets

Abstract Electricity markets have suffered important modifications in recent decades in many countries, in which a competition framework has been established with the aim of improving market efficiency and reducing energy prices. However, this new paradigm does not assure optimal solutions, as new constraints can be introduced in optimization processes that can affect the resulting prices. An example of this situation is the establishment of power purchase agreements between producers and consumers. A wide literature can be found regarding electricity markets. Some of this literature refers to the theory of spot prices and its application to them. This paper deals with the obtaining, decomposition and deduction of behavior rules of spot prices, and their influence on established contractual relationships in a deregulated market environment which allows power purchase agreements between consumers and producers. It is performed by a deterministic modeling of the complete generation-grid system. The influence of the existence of this kind of agreements on both total costs and spot prices is discussed.

Effects of submarine power transmission cables on a glass sponge reef and associated megafaunal community

We examined the effects of submarine power transmission cable installation and operation on glass sponge reef condition and associated megafauna. Video and still imagery were collected using a Remotely Operated Vehicle twice a year for 4 years following cable installation. The effects of cables on glass sponges were assessed by comparing sponge cover along fixed transects and at marked index sites. Megafauna counts along transects were used to explore the effects on associated community. We found no evidence of cable movement across the sponge reef surface. Live sponge cover was found to be consistently lower along cable transects and at cable index sites compared to controls. Live sponge cover was the lowest (55 ± 1.1% decrease) at cable index sites 1.5 years after installation and recovered to 85 ± 30.6% of the original size over the following 2 years. Our data suggest 100% glass sponge mortality along the direct cable footprint and 15% mortality in the surrounding 1.5 m corridor 3.5 years after cable installation. Growth rate of a new glass sponge was 1 and 3 cm/year in first and second year, respectively, and appeared to be seasonal. We observed a diverse megafaunal community with representatives from 7 phyla and 14 classes. Total megafauna, spot prawn, and other Arthropoda abundances were slightly lower along cable transects although the effect of cable presence was not statistically significant. The following measures could be taken to reduce the amount of damage to glass sponge reefs and associated fauna: routing the cable around reefs, whenever possible, minimizing cable movement across the surface of the reef at installation and routine operation, and assessing potential damage to glass sponges prior to decommissioning.

Shortening Delivery Times of Intensity Modulated Proton Therapy by Reducing Proton Energy Layers During Treatment Plan Optimization

To shorten delivery times of intensity modulated proton therapy by reducing the number of energy layers in the treatment plan. We have developed an energy layer reduction method, which was implemented into our in-house-developed multicriteria treatment planning system "Erasmus-iCycle." The method consisted of 2 components: (1) minimizing the logarithm of the total spot weight per energy layer; and (2) iteratively excluding low-weighted energy layers. The method was benchmarked by comparing a robust "time-efficient plan" (with energy layer reduction) with a robust "standard clinical plan" (without energy layer reduction) for 5 oropharyngeal cases and 5 prostate cases. Both plans of each patient had equal robust plan quality, because the worst-case dose parameters of the standard clinical plan were used as dose constraints for the time-efficient plan. Worst-case robust optimization was performed, accounting for setup errors of 3 mm and range errors of 3% + 1 mm. We evaluated the number of energy layers and the expected delivery time per fraction, assuming 30 seconds per beam direction, 10 ms per spot, and 400 Giga-protons per minute. The energy switching time was varied from 0.1 to 5 seconds. The number of energy layers was on average reduced by 45% (range, 30%-56%) for the oropharyngeal cases and by 28% (range, 25%-32%) for the prostate cases. When assuming 1, 2, or 5 seconds energy switching time, the average delivery time was shortened from 3.9 to 3.0 minutes (25%), 6.0 to 4.2 minutes (32%), or 12.3 to 7.7 minutes (38%) for the oropharyngeal cases, and from 3.4 to 2.9 minutes (16%), 5.2 to 4.2 minutes (20%), or 10.6 to 8.0 minutes (24%) for the prostate cases. Delivery times of intensity modulated proton therapy can be reduced substantially without compromising robust plan quality. Shorter delivery times are likely to reduce treatment uncertainties and costs.

Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis.

Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

Identification of cholesterol gallstone with two-dimensional gel electrophoresis and mass spectrometry

Background and rationale. We aimed to provide novel information to better understand the molecular mechanisms underlying gallstones formation and explore the potential protein markers for gallstones progression. The gallbladder tissues were collected from 20 patients with cholesterol gallstone and 10 liver transplant donors from November 2010 to April 2011. The proteomics were compared between gallstone patients and controls by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were identified and validated by western blotting and real-time PCR.Results. Total 19 protein spots were found to be different between two groups and 11 proteins were identified, among which 4 ones (such as Peroxiredoxin 3/Prdx3) were down-regulated and 7 (such as Tropomyosin 4/TPM4, Transgelin/SM22, Transthyretin/TTR) were up-regulated in gallstone group. Results of western blotting and RT-PCR were consistent with the 2-DE results. Conclusion. The differentially expressed proteins of TTR, TPM4, SM22 and Prdx3 may play key roles in gallstone formation and may be markers for gallstone progression.

Manganese superoxide dismutase is required to maintain osteoclast differentiation and function under static force.

Bone homeostasis is maintained by the balance of osteoblasts (OBs) and osteoclasts (OCs). Increased activity of OCs not only contributes to pathological bone resorption, such as osteoporosis and periodontitis, but also is responsible for physiological conditions like orthodontic tooth movement (OTM). However, the detailed mechanism by which orthodontic force promotes the formation of OCs is still poorly understood. In this study, we confirmed that static force promoted the differentiation of human cord monocytes (HMNCs) into OCs depending on loading time and magnitude. Protein expression profiles among HMNCs, HMNCs subjected to static force and mature OCs were established via 2-DE and MALDI-TOF-MS analyses. Total respective protein spot numbers of 549 ± 13, 612 ± 19 and 634 ± 16 were detected in each of the gels by image analysis. The five proteins identified were plasminogen activator inhibitor 2 (PAI-2, Spot 1), peroxiredoxin-6 (PRD-6, Spot 3), manganese superoxide dismutase (SOD2, Spot 6), Rho GDP-dissociation inhibitor 2 (Rho-GDI2, Spot 11) and L-lactate dehydrogenase B chain (L-LDH, Spot 15). More importantly, we revealed that SOD2 was required to maintain monocyte differentiation into functional OCs and may become a potential target for regulating the efficiency of OTM in the future.

Improvement of Lighting Uniformity and Phosphor Life in Field Emission Lamps Using Carbon Nanocoils

The lighting performances and phosphor degradation in field emission lamps (FELs) with two different kinds of cathode materials—multiwalled carbon nanotubes (MWCNTs) and carbon nanocoils (CNCs)—were compared. The MWCNTs and CNCs were selectively synthesized on 304 stainless steel wire substrates dip‐coated with nanosized Pd catalysts by controlling the growth temperature in thermal chemical vapor deposition, and the film uniformity can be optimized by adjusting the growth time. FELs were successfully fabricated by assembling these cathode filaments with a glass bulb‐type anode. The FEL with the CNC cathode showed much higher lighting uniformity and light‐spot density and a lower current at the same voltage than that with the MWCNT cathode filament, and its best luminous efficiency was as high as 75 lm/W at 8 kV. We also found that, for P22, the phosphor degradation can be effectively suppressed by replacing MWCNTs with CNCs in the cathode, due to the much larger total bright spot area and hence much lower current density loading on the anode.

Increasing proteome coverage for gel-based human tear proteome maps: towards a more comprehensive profiling.

In situations where the molecular mechanism of many ocular disorders is unknown, owing to the difficulties associated with sampling from ocular tissues, human tear film can be a promising medium in ophthalmic research. The present study demonstrates an in-depth gel-based proteome optimization survey to approach more appropriate and efficient systems in various situations such as normal and dry-eye syndromes. Therefore, systematic and statistical evaluations were performed on different preparation methods, including acetone, acetone-methanol, chloroform-methanol-water, trichloroacetic acid (TCA)-acetone, tri-n-butylphosphate-acetone-methanol precipitations and ammonium sulfate fractionation at three different percentages of saturations (50, 70 and 90%). Methods were compared quantitatively on both one- and two-dimensional patterns. Some important parameters such as total protein recovery yield, densitometric analysis of some protein contaminants, banding patterns and total spot numbers along with statistical models for proper clustering were considered. Findings revealed noticeable impacts of preparation methods on all aspects of gel-based separations as well as recovery yield (ranging from 5.29 ± 0.96 to 22.56 ± 1.77 µg/mm) and banding and pattern resolution. In addition to all these, the most important point is that the total protein spot number on the final two-dimensional patterns (varied from 528.00 ± 19.00 to 657.00 ± 21.52 for different methods) were also noticeably increased in comparison with previously published reports (maximum of 250 spots), which is essential for a more comprehensive analysis. Increasing the proteome coverage in the present study is supposed to originate from improved solubility and effective rehydration during the sample application and isoelectric focusing (IEF) procedure along with proper sample preparation.

Activity of the sun in the age of 1–2 Gyr

We discuss basic properties of the solar activity in the modern epoch and the further development of ideas about estimate the age of stars by the level of activity (gyrochronology). From comparison of properties of solar activity and processes on other G stars we have determined activity levels of the chromosphere and corona of the young Sun in its age of 1–2 Gyr. So, activity of the Sun in such an age is closest to the processes in the G stars, HD 152391 (V2292 Oph) and HD 1835 (BE Cet) with the rotational periods of 11 and 8 days correspondingly. The total spot area is of 2–3 orders of magnitude higher than the current value at the maximum of the solar cycle, the activity levels of the chromosphere and the corona are quite high and similar to the Hyades, but it did not reach the saturation level. We have analyzed new measurements of longitudinal magnetic fields and distribution of the toroidal and poloidal fields on the G main sequence stars. The mean value of the longitudinal fields for active G stars is of one order of magnitudes stronger than the mean daily magnetic field of the Sun as a star, for instance those during the 21th cycle maximum. Besides, the detection of large-scale toroidal magnetic field in such stars is important. Taking into account the activity levels and magnetic fields, we analyzed data on superflares on G stars registered with the Kepler mission. We have defined the upper limit of energy of flares that can occur in the magnetic fields of active G stars.

Applications of thin-layer chromatography in extraction and characterisation of ajoene from garlic bulbs

Novel and inexpensive methods of thin-layer chromatography (TLC) were employed for the extraction, characterisation and mechanism of quorum sensing inhibition by ajoene, a component from toluene garlic bulb (Allium sativum L.) extract (TGE). TLC profiling of TGE was carried out using ethyl acetate as solvent. Out of total spots extracted from TLC, four spots exhibited quorum sensing inhibitory (QSI) potential. Among those, spot 5 was identified as Z-ajoene by TLC and confirmed by NMR and MS. HPLC analysis indicated 97.7% purity for purified ajoene. TLC densitometric analysis quantified 221.08 μmol/g of ajoene in TGE and indicated that ajoene is stable at 4°C and at acidic pH. HPTLC profiling showed that ajoene exhibits QSI effect by inhibiting the production of both long-chain acyl homoserine lactones and Pseudomonas quinolone signal (PQS) by P. aeruginosa and also by inactivating PQS molecules.

Abstract 5193: Novel candidate oncogenes with mutation hot spots in microsatellite unstable colorectal cancer

Abstract Around 15% of colorectal cancers (CRCs) show microsatellite instability (MSI). MSI CRCs are prone to repeat mutations due to defective mismatch repair. The high background mutation frequency has discouraged systematic mutations screens in this tumor type. However, these tumors might form a sensitive system for generation and selection of oncogenic mutation hot spots. The aim of this study is to identify novel oncogenes with mutation hot spots that drive MSI CRC tumorigenesis. The exomes of 25 MSI tumors and respective healthy tissues were sequenced as the discovery set. The exome data was searched for mutation hot spots, with recurrent somatic missense mutations in at least two tumors. Potential mutation hot spots were observed in 43 genes and among these were the following known oncogenes: BRAF (V600E), CTNNB1 (T41A) and PIK3CA (H1047R). Novel potential mutation hot spots were identified in 40 genes and these were validated by Sanger sequencing. Mutation hot spots in 33 genes were confirmed and these were further screened in a validation set of 254 MSI CRCs. Fourteen genes displayed hotspot mutations also in the validation set with a total hot spot mutation frequency of 1.1-3.6 %. Many of the validated genes encode for known cancer-related proteins and for proteins with molecular and cellular functions relevant to cancer development and progression. Further work is needed to clarify the functional role of the identified hot spot mutations in MSI CRC tumorigenesis. The novel mutation hotspots may be utilized to develop personalized tumor profiling and therapy. Citation Format: Alexandra E. Gylfe, Sari Tuupanen, Ulrika Hänninen, Johanna Kondelin, Heikki Ristolainen, Riku Katainen, Esa Pitkänen, Minna Taipale, Jussi Taipale, Claus Lindbjerg Andersen, Laura Renkonen-Sinisalo, Heikki Järvinen, Jan Böhm, Jukka-Pekka Mecklin, Pia Vahteristo, Lauri A. Aaltonen. Novel candidate oncogenes with mutation hot spots in microsatellite unstable colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5193. doi:10.1158/1538-7445.AM2014-5193

Systematic production of inactivating and non-inactivating suppressor mutations at the relA locus that compensate the detrimental effects of complete spot loss and affect glycogen content in Escherichia coli.

In Escherichia coli, ppGpp is a major determinant of growth and glycogen accumulation. Levels of this signaling nucleotide are controlled by the balanced activities of the ppGpp RelA synthetase and the dual-function hydrolase/synthetase SpoT. Here we report the construction of spoT null (ΔspoT) mutants obtained by transducing a ΔspoT allele from ΔrelAΔspoT double mutants into relA+ cells. Iodine staining of randomly selected transductants cultured on a rich complex medium revealed differences in glycogen content among them. Sequence and biochemical analyses of 8 ΔspoT clones displaying glycogen-deficient phenotypes revealed different inactivating mutations in relA and no detectable ppGpp when cells were cultured on a rich complex medium. Remarkably, although the co-existence of ΔspoT with relA proficient alleles has generally been considered synthetically lethal, we found that 11 ΔspoT clones displaying high glycogen phenotypes possessed relA mutant alleles with non-inactivating mutations that encoded stable RelA proteins and ppGpp contents reaching 45–85% of those of wild type cells. None of the ΔspoT clones, however, could grow on M9-glucose minimal medium. Both Sanger sequencing of specific genes and high-throughput genome sequencing of the ΔspoT clones revealed that suppressor mutations were restricted to the relA locus. The overall results (a) defined in around 4 nmoles ppGpp/g dry weight the threshold cellular levels that suffice to trigger net glycogen accumulation, (b) showed that mutations in relA, but not necessarily inactivating mutations, can be selected to compensate total SpoT function(s) loss, and (c) provided useful tools for studies of the in vivo regulation of E. coli RelA ppGpp synthetase.

Cellular and molecular profiling of primitive HSC subsets in umbilical cord blood samples: Implications for cellular therapy

Background: Traditionally, hematopoietic stem/ progenitor cells (HSPCs) from bone-marrow, peripheral blood and cord blood donors are used to cure obstinate blood disorders and to influence tolerance in graft-versus-host disease. The most primitive HSCs in mammals comprise the well characterized CD34 and the rare disused CD34 HSCs subsets. However, the defined analysis of the molecular nature of human CD34 and their relation to CD34 HSCs fractions has been held up by the lack of specific markers. Objectives: This study was intended to provide a thorough analysis using phenotypic, functional, and systemic approaches to decipher the cellular and the molecular signature of different HSPCs components, based on a comparative study combining both surface phenotypic markers and functional ALDH activity on human cord blood samples. Materials & Methods: Different HPSCs subsets were sorted from UCB and analyzed by flow cytometry, real time PCR, and proteomic techniques and their multi-differential capacity was assessed in vitro by colony forming unit assay Results: Evaluation of different HPSCs fractions revealed that CD34LinCD38ALDH and CD34LinCD38ALDHare the most primitive HPSCs populations as illustrated by their in vitro proliferation potentials by colony forming units and stemness related gene expression. Moreover, protein profiling performed on few samples showed that there is a high degree of similarities in the total number or resolved spots in both CD34 and CD3 gels indicating that perhaps CD34are also HSCs Conclusions: Our study confirmed that HPSCs is a heterogeneous population. The simultaneous expression of surface markers and ALDH activity indicated that the neglected LinCD34CD38ALDH HPSCs compartment included primitive HSCs and displayed similar functional and molecular characteristics to their counterpart LinCD34CD38ALDH. The common protein spots identified on HSCs subsets will eventually help in their selection for scientific and clinical applications.

Improving monitoring after pancreas transplantation alone: fine-tuning of an old technique.

Graft survival after pancreas transplantation alone (PTA) is significantly poorer than graft survival after simultaneous pancreas kidney (SPK) and is particularly affected by difficulty in monitoring rejection. Exocrine bladder drainage allows assessment of pancreas graft function as urinary amylase (UA). However, standards for UA collection and interpretation are not well defined. In this study, 21 bladder-drained PTA recipients were monitored with daily values for UA and urine creatinine (Creat) concentration from post-transplant 10-mL samples and 24-h collections. Clinical events were documented and correlated to UA measurements. UA values were found to increase post-transplant until day 15, and large interpatient variability was noted (median 12676IU/L, range 668-60369IU/L). A strong correlation was found total 24-h UA production and spot UA/Creat ratio (r=0.80, p<0.001). UA/Creat ratio showed less variation during episodes of impaired renal function; therefore, urinary amylase baseline was defined as the median UA/Creat ratio after day 15. A>25% decrease of UA predicted 9/13 (69%) events. We conclude that individual baselines should be set once the values have stabilized after 15d post-transplant and that spot UA/Creat measures are reliable, patient friendly and indicate potential events after PTA.

Cross reactive cellular immune response to HCV genotype 1 and 4 antigens among genotype 4 exposed subjects.

BackgroundHepatitis C Virus (HCV) infection is a global health burden particularly in Egypt, where HCV genotype 4a (GT-4a) predominates. The prevention and control of HCV infection will remain a challenge until the development of an effective vaccine that protects against different genotypes. Several HCV GT-1-based vaccines are in different stages of clinical trials, but antigenic differences could make protection against other genotypes problematic. In this regard, data comparing the cell-mediated immune (CMI) response to different HCV genotypes are limited. We aimed to ex vivo investigate whether GT-1-based vaccine may protect against HCV GT-4 infections. This was carried out on samples collected from genotype 4 infected/exposed subjects.Methods/Principal FindingsThe CMI responses of 35 subjects; infected with HCV GT-4/or who had spontaneously-resolved the infection and 10 healthy control subjects; to two sets of seven HCV overlapping 15-mer peptide pools derived from both genotypes; and covering most of the viral proteins; were evaluated. This was carried out using an interferon gamma (IFNγ) enzyme-linked immunospot (ELISpot) assay. Peripheral blood mononuclear cells (PBMC) from 17 subjects (48%) responded to at least one peptide pool derived from GT-1b/GT-4a with 13 subjects responding to peptide pools from both genotypes. A strong correlation was found in the responses to both genotypes (r = 0.82, p<0.001; 95% confidence interval = 0.562–0.933). The average IFNγ total spot forming cells (SFC)/106 PBMC (±SE) from the responding subjects for GT-1b and GT-4a was 216±56 and 199±55, respectively (p = 0.833). Also, there were no significant differences between those who cleared their HCV infection or who remained HCV-RNA positive (p = 0.8).Conclusion/SignificanceOur data suggest that an effective GT-1b vaccine could protect from GT-4a infection. These data could help in HCV rationale vaccine design and efficacy studies and further our understanding of HCV cross protection against different genotypes.

UJI KETAHANAN BEBERAPA GENOTIPE TANAMAN KARET TERHADAP PENYAKIT Corynespora cassiicola DAN Colletotrichum gloeosporioides DI KEBUN ENTRES SEI PUTIH

Research on title resistence test of rubber plant genotype by Corynespora cassiicola andColletotrichum gloeosporioides diseases at Sungei Putih budstock rubber plantation. This researchwas to study to determine resistence of rubber plant genotype against C. cassiicola andC. gloeosporioides at budstock rubber plantation. This research had been conducted atexperimental field of Sungei Putih Rubber Research Center in May-July 2013 using split plotdesign with two factor, i.e genotype plants (930, 135, 38, 51, 65, 100, 108, 118, 222, 223, 227, and374) and diseases type (no disease, C. cassiicola and C. gloeosporioides). The results showed thatthe fastest incubation periode (3 days) contained on genotype 223 and 930, the slowest (6 days) ongenotype 222. The lowest disease severity (20,99%) of C. cassiicola treatment contained ongenotype 100 and the highest (33,33%) was at genotype 65. The lowest disease severity (21,61%)of C. gloeosporioides treatment contained on genotype 222 and the highest (57,41%) was ongenotype 223. The highest total spot (58,56 spots) contained on combination treatment of genotype223 with C. gloeosporioides disease and the lowest (1,70 spots) on combination treatment ofgenotype 100 with C. cassiicola disease.Key Words: Genotype plant rubber, Corynespora cassiicola, Colletotrichum gloeosporioides

Proteomic analysis of the sheep caruncular and intercaruncular endometrium reveals changes in functional proteins crucial for the establishment of pregnancy

The expression and regulation of endometrial proteins are crucial for conceptus implantation and development. However, little is known about site-specific proteome profiles of the mammalian endometrium during the peri-implantation period. We utilised a two-dimensional gel electrophoresis/mass spectrometry-based proteomics approach to compare and identify differentially expressed proteins in sheep endometrium. Caruncular and intercaruncular endometrium were collected on days 12 (C12) and 16 (C16) of the oestrous cycle and at three stages of pregnancy corresponding to conceptus pre-attachment (P12), implantation (P16) and post-implantation (P20). Abundance and localisation changes in differentially expressed proteins were determined by western blot and immunohistochemistry. In caruncular endometrium, 45 protein spots (5% of total spots) altered between day 12 of pregnancy (P12) and P16 while 85 protein spots (10% of total spots) were differentially expressed between P16 and C16. In intercaruncular endometrium, 31 protein spots (2% of total spots) were different between P12 and P16 while 44 protein spots (4% of total spots) showed differential expression between C12 and C16. The pattern of protein changes between caruncle and intercaruncle sites was markedly different. Among the protein spots with implantation-related changes in volume, 11 proteins in the caruncular endometrium and six proteins in the intercaruncular endometrium, with different functions such as protein synthesis and degradation, antioxidant defence, cell structural integrity, adhesion and signal transduction, were identified. Our findings highlight the different but important roles of the caruncular and intercaruncular proteins during early pregnancy.

The influence of the Children’s Food and Beverage Advertising Initiative: change in children’s exposure to food advertising on television in Canada between 2006–2009

To assess whether children's exposure to television food/beverage advertising has changed since the implementation of the self-regulatory Canadian Children's Food and Beverage Advertising Initiative (CAI). Data on 11 advertised food/beverage categories (candy, chocolate bars, cookies, portable snacks, cheese, yogurt, cereal, juices, soft drinks, diet soft drinks and fast food) were purchased from Nielsen Media Research for May 2006, 2009 and 2011 for the broadcasting markets of Toronto and Vancouver. The number of advertisements aired on 27 television stations between 0600 hours-1200 hours was determined in Toronto and Vancouver for May 2006, 2009 and 2011 and the percentage change in the number of spots between May 2006 and May 2011 on all stations, on children's specialty stations and on generalist stations was then calculated. The average number food/beverage spots seen by children aged 2-11 was determined for May 2006 and 2009 and the percentage change was calculated. On children's specialty channels, a 4.5% decrease in total spots aired was observed while spots aired on generalist stations increased by 44% (Toronto) and 45% (Vancouver). On all stations, children's total average exposure to food/beverage advertising increased by 16.8% in Toronto and 6.4% in Vancouver between 2006 and 2009. Significant increases were seen in snacks and yogurt in both cities, and in fast food in Toronto. On children's specialty channels, children's exposure to the food/beverage categories considered increased by 5.4% in Toronto and by 2.5% in Vancouver. Despite improvements in the volume of spots on children's specialty channels, children's exposure to food and beverage advertising has increased since the implementation of the CAI. The current self-regulatory system is failing to protect children from food marketing high in fat, sugar and sodium on television. Government regulation needs to be considered.

Hepatitis C virus-specific T-cell response correlates with hepatitis activity and donor IL28B genotype early after liver transplantation.

It is not known how the immune system targets hepatitis C virus (HCV)-infected HLA-mismatched hepatocytes under immune-suppressed conditions after orthotopic liver transplantation (OLT). In addition, the relationship between the HCV-specific immune response and IL28B variants as predictors of HCV clearance has not been well-characterized. We determined the IL28B polymorphisms for 57 post-OLT HCV carriers, and we assessed the HCV-specific immune responses by measuring the peripheral blood mononuclear cell-derived HCV-specific interferon-gamma (IFN-γ) response using an enzyme-linked immunospot assay. At 1-3 years after OLT, patients with no active hepatitis showed higher total spots on the immunospot assay. At＞3 years after OLT, patients with resolved HCV showed higher levels of core, NS3, NS5A, and total spots compared to the chronic hepatitis patients. The IL28B major genotype in the donors correlated with higher spot counts for NS5A and NS5B proteins at 1-3 years after OLT. In the post-OLT setting, the HCV-specific immune response could be strongly induced in patients with no active hepatitis with an IL28B major donor or sustained virological response. Strong immune responses in the patients with no active hepatitis could only be maintained for 3 years and diminished later. It may be beneficial to administer IFN treatment starting 3 years after OLT, to induce the maximum immunological effect.