Minimum inhibitory concentrations of frequently used antibiotics against Escherichia coli and Trueperella pyogenes isolated from uteri of postpartum dairy cows.

The objective of this study was to determine minimum inhibitory concentrations (MIC) of frequently used antimicrobials for Escherichia coli and Trueperella pyogenes isolated from postpartum bovine uteri of cows with acute puerperal metritis (APM, n = 67), cows suspected to have APM (n = 37), and healthy cows (n = 37) and to evaluate possible differences in MIC according to clinical signs. Cows with APM had reddish-brown, fetid vaginal discharge and rectal temperature (RT) ≥39.5°C within 21 d in milk; cows suspected to have APM had either reddish-brown, fetid vaginal discharge or RT ≥39.5°C within 21 d in milk; and healthy cows had neither fetid discharge nor RT ≥39.5°C. Samples were collected from cows on commercial dairy herds (n = 7) using the cytobrush technique. A total of 37 T. pyogenes isolates and 85 E. coli isolates were tested. Ceftiofur, a third-generation cephalosporin that is often used to treat APM, was the focus of analysis. Trueperella pyogenes and E. coli were isolated more often from samples of cows with APM (46 and 90%, respectively) compared with samples from healthy cows (19 and 54%, respectively). Regarding cows suspected to have APM, T. pyogenes and E. coli were numerically more often isolated (30 and 70%, respectively) than in healthy cows (19 and 54%, respectively). Minimum inhibitory concentrations of ceftiofur were low. For T. pyogenes and E. coli, MIC50 (concentration that inhibited growth of 50% of isolates) were 0.25 and 0.5 µg/mL and MIC90 (concentration that inhibited growth of 90% of isolates) were 0.5 and 1 µg/mL, respectively. Although ceftiofur inhibited all T. pyogenes at the highest concentration tested (64 µg/mL), the growth of 5.9% of E. coli was not impaired. Recently, ampicillin has been suggested as an alternative treatment for APM. Although the T. pyogenes isolates exhibited low MIC in general (MIC50 ≤0.015 µg/mL and MIC90 = 0.06 µg/mL) and 81.1% of all T. pyogenes could be inhibited at the lowest ampicillin concentration tested, 11.8% of the E. coli isolates were not impaired at the highest concentration (64 µg/mL) tested in this study. The MIC50 and MIC90 of E. coli were 4 and ≥128 µg/mL, respectively. We detected no difference in the MIC distributions of ceftiofur or ampicillin among isolates from the 3 APM groups. In summary, E. coli with high MIC against ceftiofur as well as against ampicillin were found in this study.

Short communication: Effect of adding a second prostaglandin F2α injection during the Ovsynch protocol on luteal regression and fertility in lactating dairy cows: A meta-analysis.

Incomplete luteal regression after treatment with a single dose of prostaglandin F2α during an Ovsynch protocol decreases fertility to timed artificial insemination (TAI). To increase the proportion of cows with complete luteal regression and subsequently pregnancy per artificial insemination (P/AI), an additional treatment with PGF2α 24 h after the first has been recommended. A systematic review of the literature and meta-analytical assessment were performed with the objective of evaluating the effects of adding a second PGF2α treatment during the Ovsynch protocol on luteal regression and reproductive performance in lactating dairy cows. Based on the heterogeneity among the experimental treatments, a fixed or a random effects meta-analysis was conducted. Reproductive outcomes of interest were luteal regression at the end of the Ovsynch protocol, and P/AI measured 32 to 39 d after TAI. Seven randomized controlled experiments from 6 published manuscripts including 5,356 cows with the primary objective to evaluate the effect of an additional treatment with PGF2α during the Ovsynch protocol on P/AI were used. Information regarding luteal regression at the end of the Ovsynch protocol was available for 1,856 cows. Adding a second PGF2α treatment on d 8 during the Ovsynch protocol increased the relative risk (RR) of complete luteal regression at the end of the Ovsynch protocol (RR = 1.14; 95% confidence interval = 1.10 to 1.17) using a fixed effects model and the RR for pregnancy (RR = 1.14; 95% confidence interval = 1.06 to 1.22) 32 d after TAI using a fixed effects model. No heterogeneity was observed among the 6 manuscripts regarding complete luteal regression and P/AI. In summary, there was a clear benefit of an additional PGF2α treatment during the Ovsynch protocol on luteal regression (+11.6 percentage units) and on P/AI (+4.6 percentage units).

Evaluation of prostaglandin F2α versus prostaglandin F2α plus gonadotropin-releasing hormone as Presynch methods preceding an Ovsynch in lactating dairy cows: A meta-analysis.

Presynchronization of cows with 2 injections of prostaglandin administered 14 d apart (Presynch-Ovsynch) is a widely adopted procedure to increase pregnancy per artificial insemination (P/AI) at first service. Recently, a presynchronization protocol including GnRH and PGF2α (Double-Ovsynch; GnRH, 7 d, PGF2α, 3 d, GnRH) followed 7 d later by an Ovsynch protocol was introduced to overcome the limitations of PGF2α-based protocols for presynchronization of anovular cows and to precisely set up cows on d 7 of the estrous cycle when the Ovsynch is initiated. A systematic review of the literature and a meta-analytical assessment was performed with the objective to compare the reproductive performance of lactating dairy cows presynchronized with these 2 protocols for the first timed AI (TAI) considering parity-specific effects. A fixed or a random effects meta-analysis was used based on the heterogeneity among the experimental groups. Reproductive outcomes of interest were P/AI measured on d 32 (28-42) and pregnancy loss between d 32 and 60 (42-74) of gestation. A total of 25 articles with 27 experimental groups from 63 herds including 21,046 cows submitted to first TAI using either a Presynch-Ovsynch or a Double-Ovsynch protocol were reviewed. Results for P/AI were then categorized by parity if available. Information was available for P/AI for 7,400 and 10,999 primiparous and multiparous cows, respectively. Information regarding pregnancy loss was available for 7,477 cows. In the random effects model for all cows, the overall proportion of P/AI was 41.7% [95% confidence interval (CI): 39.1-44.3; n = 8,213] and 46.2% (95% CI: 41.9-50.5; n = 12,833) on d 32 after TAI for Presynch-Ovsynch and Double-Ovsynch, respectively. In the random effects model for primiparous cows, the overall proportion of P/AI was 43.4% (95% CI: 36.2-47.7; n = 2,614) and 51.4% (95% CI: 47.4-55.4; n = 4,786) on d 32 after TAI for Presynch-Ovsynch and Double-Ovsynch, respectively. In the random effects model for multiparous cows, the overall proportion of P/AI was 39.2% (95% CI: 36.2-42.3; n = 3,411) and 41.4% (95% CI: 36.4-46.4; n = 7,588) on d 32 after TAI for Presynch-Ovsynch and Double-Ovsynch, respectively. The overall proportion of pregnancy loss was 11.3% (95% CI: 7.6-15.7; n = 3,247) and 11.7% (95% CI: 9.3-14.3; n = 4,230) on d 60 after AI for Presynch-Ovsynch to and Double-Ovsynch, respectively. Substantial heterogeneity existed among the experimental groups regarding P/AI and pregnancy loss. In summary, a benefit was detected for P/AI in primiparous cows presynchronized with a Double-Ovsynch protocol for the first TAI, but this benefit was not observed in multiparous cows.

Laparoscopic Adrenalectomy is Feasible for Suspected Adrenal Tumors in Children Younger than 24 Months of Age - But is it Always Justified?

INTRODUCTION: Tumors of the adrenal gland are rare in children younger than 24 months of age. While neuroblastomas are most important in this age group, adrenal hemorrhage and other tumors are sometimes difficult to distinguish. Harvesting biopsies is mandatory in these young patients to obtain information on molecular markers, namely, MYCN and 1p deletion. PATIENTS: Between 03/2012 and 10/2013, 11 patients younger than 24 months of age with suspicious adrenal tumors were operated on laparoscopically. METHODS: The diagnostic workup was coordinated by our pediatric oncologists according to the terms of the NB2004 trial protocol. RESULTS: 9 out of 11 had a diagnosis of neuroblastoma, the others were adenoma respective complete necrosis of the adrenal gland. All of the neuroblastomas were negative for both MYCN amplification and 1p deletion. A complete resection was successful in 9 out of 11 cases. 3 complications occurred, 1 major and 2 minor. DISCUSSION AND CONCLUSION: Behind the recognition that laparoscopic adrenalectomy is technically feasible, the fact that all neuroblastomas were negative for MYCN amplification and 1p deletion raises the issue of whether biopsy is mandatory for risk stratification in this age group.

Image-defined Risk Factors Correlate with Surgical Radicality and Local Recurrence in Patients with Neuroblastoma.

BACKGROUND: Neuroblastoma is the second most common solid pediatric tumor and the most common cancer to be detected in children younger than 12 months of age. To date, 2 different staging systems describe the extent of the disease: the International Neuroblastoma Staging System (INSS) and the International Neuroblastoma Risk Group Staging System (INRGSS). The INRGSS-system is characterized by the presence or absence of so called image-defined risk factors (IDRFs), which are described as surgical risk factors. We hypothesized that IDRFs correlate with surgical complications, surgical radicality, local recurrence and overall survival (OS). PATIENTS AND METHODS: Between 2003 and 2010, 102 patients had neuroblastoma surgery performed in our department. We analyzed medical records for IDRF-status and above named data. RESULTS: 16 patients were IDRF-negative, whereas 86 patients showed one or more IDRF. Intra- or postoperative complications have been reported in 21 patients (21%). 19 of them showed one or more IDRF and 2 patients were IDRF-negative (p=n.s.). Patients who suffered from intra- or postoperative complications demonstrated a decreased OS (p=0.011). Statistical analysis revealed an inverse correlation between the extent of macroscopical removal and IDRF-status (p=0.001). Furthermore, the number of IDRFs were associated with a decreased likelihood of radical tumor resection (p<0.001). 19 patients had local recurrence; all of them were IDRF-positive (p=0.037). CONCLUSIONS: Pediatric surgeons should consider IDRFs as a useful tool for risk assessment and therefore planning for neuroblastoma surgery.

Randomized, controlled clinical trial on the efficacy of nonsteroidal antiinflammatory drugs for the treatment of acute puerperal metritis in dairy cows.

The objective of this study was to assess the efficacy of ketoprofen compared with ceftiofur hydrochloride for the treatment of acute puerperal metritis (APM). Specifically, we set out to compare the incidence of extended treatment (extT) between treatment groups, to determine the prevalence of purulent vaginal discharge (PVD) and milk yield on the first 3 milk tests postpartum, and to analyze reproductive performance of cows treated with ketoprofen or ceftiofur. Cows with rectal temperature ≥39.5°C and reddish-brown fetid vaginal discharge within the first 10 d in milk (DIM) were diagnosed with APM. Day of enrollment and first day of treatment was considered study day 1. Rectal temperature was recorded daily until study day 7. A total of 610 dairy cows with APM were enrolled in this randomized clinical trial. Cows meeting the inclusion criteria were allocated to treatment with ketoprofen (3mg/kg of body weight, n=300) or treatment with ceftiofur (1mg/kg of body weight, n=310) on study days 1, 2, and 3. Cows that showed rectal temperature ≥39.5°C between study days 4 and 7 received an extT with ceftiofur for 3 (ketoprofen group) or 2 (ceftiofur group) more days. Cows were examined with the Metricheck device (Simcro, Hamilton, New Zealand) between DIM 21 and 40, and vaginal discharge was categorized on a 5-point scale according to the presence of pus. Cows with a score ≥2 were classified as having PVD. Fifty-two cows (35 from ketoprofen group, 17 from ceftiofur group) were excluded from analysis due to missing protocol compliance (n=37) or concurrent disease (n=15). Cows treated with ketoprofen were more likely to have an extT than cows treated with ceftiofur (61 vs. 31%). Prevalence of PVD did not differ between the 2 treatment groups (ketoprofen, 56%; ceftiofur, 53%). Cows, however, that needed an extT after the initial 3-d treatment were more likely to develop PVD than cows without extT (64 vs. 46%). Treatment group did not affect milk yield (ketoprofen group, 35.5±0.4kg; ceftiofur group, 35.2±0.3kg), first artificial insemination pregnancy risk (ketoprofen group, 20% vs. ceftiofur group, 25%), median days to first artificial insemination [ketoprofen group, 73 d, 95% confidence interval (CI): 70-75 d vs. ceftiofur group, 75 d, 95% CI: 72-76 d] and median days to pregnancy (ketoprofen group, 144 d, 95% CI: 132-158 d vs. ceftiofur group, 133 d, 95% CI: 119-153 d). These results indicate that although cows initially treated with ketoprofen were more likely to receive extT, fewer doses of ceftiofur (1.83) were required compared with cows initially treated with ceftiofur (3.63). Moreover, the prevalence of PVD was not increased and milk yield and reproductive performance were not negatively affected by the initial treatment with ketoprofen.

Photoinduced reduction of surface states in Fe:ZnO.

We report on the electronic structure of nano-crystalline Fe:ZnO, which has recently been found to be an efficient photocatalyst. Using resonant photoemission spectroscopy, we determine the binding energy of Fe 3d states corresponding to different valencies and coordination of the Fe atoms. The photo-activity of ZnO reduces Fe from 3+ to 2+ in the surface region of the nano-crystalline material due to the formation of oxygen vacancies. Electronic states corresponding to low-spin Fe(2+) are observed and attributed to crystal field modification at the surface. These states are potentially important for the photocatalytic sensitivity to visible light due to their location deep in the ZnO bandgap. X-ray absorption and x-ray photoemission spectroscopy suggest that Fe is only homogeneously distributed for concentrations up to 3%. Increased concentrations does not result in a higher concentration of Fe ions in the surface region. This is limiting the photocatalytic functionality of ZnO, where the most efficient Fe doping concentration has been shown to be 1%-4%.

The associations between postpartum serum haptoglobin concentration and metabolic status, calving difficulties, retained fetal membranes, and metritis.

Measurement of serum haptoglobin (Hapto) concentrations results in only moderate reported specificity and sensitivity for the detection of metritic cows. The objective of this study was to evaluate associations between different variables and haptoglobin concentrations after calving. Parity, periparturient metabolic stress indicated by ß-hydroxybutyric acid (BHBA) and nonesterified fatty acids (NEFA), calving difficulties, retained fetal membranes (RFM), and acute puerperal metritis (APM) were evaluated. A total of 443 Holstein Friesian cows were enrolled in this retrospective observational study. Acute puerperal metritis was diagnosed when a cow had fetid, reddish-brown, watery vaginal discharge in combination with rectal temperature ≥39.5°C. The retention of the fetal membranes has been defined as the failure to expel the fetal membranes within 12h after calving. Results of blood samples from 2 and 5 d in milk (DIM) were analyzed for Hapto, BHBA, and NEFA. Primiparous cows had a greater median Hapto concentration than multiparous cows at 5 DIM [primiparous: 2.25g/L, interquartile range (IQR) 1.45-2.50, n=146; multiparous: 1.13g/L, IQR 0.52-2.22, n=302; P<0.05]. Therefore, different Hapto thresholds based on references from literature for all cows (1.4g/L), primiparous cows (2.49g/L), and multiparous cows (1.4g/L) were used for further analysis. Periparturient metabolic stress indicated by elevated BHBA (≥1.2mmol/L) at 5 DIM was associated with elevated Hapto (odds ratio 2.39-2.87) regardless of parity. In contrast, elevated NEFA (≥0.6mmol/L) at 2 DIM was not a risk factor for elevated Hapto. Multiparous cows with assisted calving had a 2.46 times higher risk for elevated Hapto, whereas primiparous cows with assisted calving had no elevated risk for elevated Hapto at 5 DIM. Moreover, multiparous cows with RFM were 5.51 times more likely to have elevated Hapto at 5 DIM than cows without RFM. Acute puerperal metritis within the first 5 DIM was associated with elevated Hapto (odds ratio 2.74-5.01), regardless of parity. We speculate that the association of calving ease, RFM, and periparturient metabolic stress could explain the moderate sensitivity and specificity reported for the detection of metritic cows by measuring Hapto.

Technical note: Assessment of milk temperature measured by automatic milking systems as an indicator of body temperature and fever in dairy cows.

The objective of this study was to evaluate whether milk temperature (MT) measured by automatic milking system (AMS) is a reliable indicator of body temperature of dairy cows and whether cows with fever could be detected. Data loggers (Minilog 8, Vemco Ltd., Halifax, NS, Canada) measuring body temperature were inserted for 7 ± 1 d into the vaginal cavity of 31 dairy cows and programmed to take 1 reading/min. Milk temperature was recorded at each milking event by the AMS, and values from the vaginal loggers were paired with the corresponding MT. The correlation (r) between vaginal temperature (VT) and MT was 0.52. Vaginal temperature was higher (39.1 ± 0.4°C) than MT (38.6 ± 0.7°C) with a mean difference of 0.5 ± 0.6°C. The ability of MT to identify cows with fever was assessed using 2 approaches. In the first approach, VT could indicate fever at any time of the day, whereas MT could display fever only during the milking events of a given day. Different definitions of fever based on thresholds of VT and duration exceeding these thresholds were constructed. Different thresholds of MT were tested to distinguish between cows with and without fever. The combination of 39.0°C as a threshold for MT and 39.5°C for at least 2h/d as a threshold for VT resulted in the highest combination of sensitivity (0.65) and specificity (0.65). In the second approach, we evaluated whether MT could identify cows with fever at a given milking event. A threshold of MT >38.7°C delivered the best combination of sensitivity (0.77) and specificity (0.66) when fever was defined as VT ≥39.5°C. Therefore, MT measured by AMS can be indicative of fever in dairy cows to a limited extent.

Pharmacogenetic profiling of CD133 is associated with response rate (RR) and progression-free survival (PFS) in patients with metastatic colorectal cancer (mCRC), treated with bevacizumab-based chemotherapy.

Recent studies suggest CD133, a surface protein widely used for isolation of colon cancer stem cells, to be associated with tumor angiogenesis and recurrence. We hypothesized that gene expression levels and germline variations in CD133 will predict clinical outcome in patients with metastatic colorectal cancer (mCRC), treated in first-line setting with 5-fluorouracil, oxaliplatin and bevacizumab (BV), and we investigated whether there is a correlation with gene expression levels of CD133, vascular endothelial growth factor (VEGF) and its receptors. We evaluated intra-tumoral gene expression levels by quantitative real-time (RT) PCR from 54 patients and three germline variants of the CD133 gene by PCR-restriction-fragment length polymorphism from 91 patients with genomic DNA. High gene expression levels of CD133 (>7.76) conferred a significantly greater tumor response (RR=86%) than patients with low expression levels (7.76, RR=38%, adjusted P=0.003), independent of VEGF or its receptor gene expression levels. Gene expression levels of CD133 were significantly associated with VEGF and its receptors messenger RNA levels (VEGFR-1 (P<0.01), -2 and -3, P<0.05). Combined analyses of two polymorphisms showed a significant association with progression-free survival (PFS) (18.5 months vs 9.8 months, P=0.004) in a multivariate analysis as an independent prognostic factor for PFS (adjusted P=0.002). These results suggest that CD133 is a predictive marker for standard first-line BV-based treatment in mCRC.

The structure, thermal properties and phase transformations of the cubic polymorph of magnesium tetrahydroborate.

The structure of the cubic polymorph of magnesium tetrahydroborate (γ-Mg(BH(4))(2)) has been determined in space group Ia3d from a structural database of the isoelectronic compound SiO(2); this has been corroborated by DFT calculations. The structure is found to concur with that recently determined by Filinchuk et al. (Y. Filinchuk, B. Richter, T. R. Jensen, V. Dmitriev, D. Chernyshov and H. Hagemann, Angew. Chem. Int. Ed., 2011, DOI: 10.1002/anie.201100675). The phase transformations and subsequent decomposition of γ-Mg(BH(4))(2) on heating have been ascertained from variable-temperature synchrotron X-ray diffraction data combined with thermogravimetric and mass spectrometry measurements. At ~160 °C, conversion to a disordered variant of the ß-Mg(BH(4))(2) phase (denoted as ß') is observed along with a further unidentified polymorph. There is evidence of amorphous phases during decomposition but there is no direct crystallographic indication of the existence of Mg(B(12)H(12)) or other intermediate Mg-B-H compounds. MgH(2) and finally Mg are observed in the X-ray diffraction data after decomposition.

Minimally invasive surgery for neuroblastic tumours: A SIOPEN multicentre study: Proposal for guidelines.

INTRODUCTION: Surgery plays a key role in the management of Neuroblastic tumours (NB), where the standard approach is open surgery, while minimally invasive surgery (MIS) may be considered an option in selected cases. The indication(s) and morbidity of MIS remain undetermined due to small number of reported studies. The aim of this study was to critically address the contemporary indications, morbidity and overall survival (OS) and propose guidelines exploring the utility of MIS for NB. MATERIALS & METHODS: A SIOPEN study where data of patients with NB who underwent MIS between 2005 and 2018, including demographics, tumour features, imaging, complications, follow up and survival, were extracted and then analysed. RESULTS: A total of 222 patients from 16 centres were identified. The majority were adrenal gland origin (54%) compared to abdominal non-adrenal and pelvic (16%) and thoracic (30%). Complete and near complete macroscopic resection (>95%) was achieved in 95%, with 10% of cases having conversion to open surgery. Complications were reported in 10% within 30 days of surgery. The presence of IDRF (30%) and/or tumour volume >75 ml were risk factors for conversion and complications in multivariate analysis. Overall mortality was 8.5%. CONCLUSIONS: MIS for NB showed that it is a secure approach allowing more than 95% resection. The presence of IDRFs was not an absolute contraindication for MIS. Conversion to open surgery and overall complication rates were low, however they become significant if tumour volume >75 mL. Based on these data, we propose new MIS guidelines for neuroblastic tumours.

A let-7 microRNA-binding site polymorphism in 3'-untranslated region of KRAS gene predicts response in wild-type KRAS patients with metastatic colorectal cancer treated with cetuximab monotherapy.

PURPOSE: recent studies have found that KRAS mutations predict resistance to monoclonal antibodies targeting the epidermal growth factor receptor in metastatic colorectal cancer (mCRC). A polymorphism in a let-7 microRNA complementary site (lcs6) in the KRAS 3' untranslated region (UTR) is associated with an increased cancer risk in non-small-cell lung cancer and reduced overall survival (OS) in oral cancers. We tested the hypothesis whether this polymorphism may be associated with clinical outcome in KRAS wild-type (KRASwt) mCRC patients treated with cetuximab monotherapy. PATIENTS AND METHODS: the presence of KRAS let-7 lcs6 polymorphism was evaluated in 130 mCRC patients who were enrolled in a phase II study of cetuximab monotherapy (IMCL-0144). Genomic DNA was extracted from dissected formalin-fixed paraffin-embedded tumor tissue, KRAS mutation status and polymorphism were assessed using direct sequencing and PCR restriction fragment length polymorphism technique. RESULTS: KRAS let-7 lcs6 polymorphism was found to be related to object response rate (ORR) in mCRC patients whose tumors had KRASwt. The 12 KRASwt patients harboring at least a variant G allele (TG or GG) had a 42% ORR compared with a 9% ORR in 55 KRASwt patients with let-7 lcs6 TT genotype (P = 0.02, Fisher's exact test). KRASwt patients with TG/GG genotypes had trend of longer median progression-free survival (3.9 versus 1.3 months) and OS (10.7 versus 6.4 months) compared to those with TT genotypes. CONCLUSIONS: these results are the first to indicate that the KRAS 3'UTR polymorphism may predict for cetuximab responsiveness in KRASwt mCRC patients, which warrants validation in other clinical trials.

Pharmacogenetic profiling of Aurora kinase B is associated with overall survival in metastatic colorectal cancer.

Aurora kinases are conserved eukaryotic serine-threonine kinases, which serve as key regulators of mammalian mitosis. Several studies revealed a distinct correlation between inaccurate chromosome segregation, leading to chromosomal number instability, cancer progression and poor outcome. The aim of this study was to investigate the correlation of Aurora kinases A (AURKA) and B (AURKB) with overall survival (OS) by quantifying gene expression analysis and evaluation of single-nucleotide polymorphisms (SNPs) in human colorectal cancer samples and assessing the associations with clinicopathological features. We evaluated intratumoral gene expression levels and SNPs of AURKA and -B from 41 patients with metastatic colorectal cancer (mCRC). Patients with a high expression level of AURKB (>1.28) lived significantly shorter (n=11, median OS=6.4 months, 95% confidence interval (CI): 3.0-14.5 months) compared with patients with a low expression level (≤ 1.28) (n=30, median OS=18.4 months, 95% CI: 14.7-27.8 months, P=0.026, Wald's test). Patients harboring any G-allele in AURKB 885A>G showed a significantly decreased OS (P=0.05, log-rank test). We did not find any associations with clinicopathological variables and AURKA gene expression levels. Our results suggest a potential role for AURKB inhibition in patients with mCRC; thereby supporting its potential role as a target in mCRC.

The UCSC Genome Browser Database: update 2009.

The UCSC Genome Browser Database (GBD, http://genome.ucsc.edu) is a publicly available collection of genome assembly sequence data and integrated annotations for a large number of organisms, including extensive comparative-genomic resources. In the past year, 13 new genome assemblies have been added, including two important primate species, orangutan and marmoset, bringing the total to 46 assemblies for 24 different vertebrates and 39 assemblies for 22 different invertebrate animals. The GBD datasets may be viewed graphically with the UCSC Genome Browser, which uses a coordinate-based display system allowing users to juxtapose a wide variety of data. These data include all mRNAs from GenBank mapped to all organisms, RefSeq alignments, gene predictions, regulatory elements, gene expression data, repeats, SNPs and other variation data, as well as pairwise and multiple-genome alignments. A variety of other bioinformatics tools are also provided, including BLAT, the Table Browser, the Gene Sorter, the Proteome Browser, VisiGene and Genome Graphs.

Stressed volume estimated by finite element analysis predicts the fatigue life of human cortical bone: The role of vascular canals as stress concentrators.

The fatigue life of cortical bone can vary several orders of magnitude, even in identical loading conditions. A portion of this variability is likely related to intracortical microarchitecture and the role of vascular canals as stress concentrators. The size, spatial distribution, and density of canals determine the peak magnitude and volume of stress concentrations. This study utilized a combination of experimental fatigue testing and image-based finite element (FE) analysis to establish the relationship between the stressed volume (i.e., volume of bone above yield stress) associated with vascular canals and the fatigue life of cortical bone. Thirty-six cortical bone samples were prepared from human femora and tibiae from five donors. Samples were allocated to four loading groups, corresponding to stress ranges of 60, 70, 80, and 90 MPa, then cyclically loaded in zero-compression until fracture. Porosity, canal diameter, canal separation, and canal number for each sample was quantified using X-ray microscopy (XRM) after testing. FE models were created from XRM images and used to calculate the stressed volume. Stressed volume was a good predictor of fatigue life, accounting for 67% of the scatter in fatigue-life measurements. An increase in stressed volume was most strongly associated with higher levels of intracortical porosity and larger canal diameters. The findings from this study suggest that a large portion of the fatigue-life variance of cortical bone in zero-compression is driven by intracortical microarchitecture, and that fatigue failure may be predicted by quantifying the stress concentrations associated with vascular canals.

IL-6 deficiency in mice neither impairs induction of metabolic genes in the liver nor affects blood glucose levels during fasting and moderately intense exercise.

AIMS/HYPOTHESIS: Fasting and exercise are strong physiological stimuli for hepatic glucose production. IL-6 has been implicated in the regulation of gluconeogenic genes, but the results are contradictory and the relevance of IL-6 for fasting- and exercise-induced hepatic glucose production is not clear. METHODS: Investigations were performed in rat hepatoma cells, and on C57Bl6 and Il6(-/-) mice under the following conditions: IL-6 stimulation/injection, non-exhaustive exercise (60 min run on a treadmill) and fasting for 16 h. Metabolite analysis, quantitative real-time PCR and immunoblotting were performed. RESULTS: IL-6 stimulation of rat hepatoma cells led to higher glucose production. Injection of IL-6 in mice slightly increased hepatic Pepck (also known as Pck1) expression. Fasting of Il6(-/-) mice for 16 h did not alter glucose production compared with wild-type mice, since plasma glucose concentrations were similar and upregulation of phosphoenolpyruvate carboxykinase (PEPCK) and Pgc-1alpha (also known as Ppargc1a) expression was comparable. In the non-fasting state, Il6(-/-) mice showed a mild metabolic alteration including higher plasma glucose and insulin levels, lower NEFA concentrations and slightly increased hepatic PEPCK content. Moderately intense exercise resulted in elevated IL-6 plasma levels in wild-type mice. Despite that, plasma glucose, insulin, NEFA levels and hepatic glycogen content were not different in Il6(-/-) mice immediately after running, while expression of hepatic G6pc, Pgc-1alpha, Irs2 and Igfbp1 mRNA was similarly increased. CONCLUSIONS/INTERPRETATION: These data suggest that in mice IL-6 is not essential for physiologically increased glucose production during fasting or non-exhaustive exercise.

Activation of the mitogen-activated protein kinase (MAPK) signalling pathway in the liver of mice is related to plasma glucose levels after acute exercise.

AIMS/HYPOTHESIS: We aimed to identify, in the liver of mice, signal transduction pathways that show a pronounced regulation by acute exercise. We also aimed to elucidate the role of metabolic stress in this response. METHODS: C57Bl6 mice performed a 60 min run on a treadmill under non-exhaustive conditions. Hepatic RNA and protein lysates were prepared immediately after running and used for whole-genome-expression analysis, quantitative real-time PCR and immunoblotting. A subset of mice recovered for 3 h after the treadmill run. A further group of mice performed the treadmill run after having received a vitamin C- and vitamin E-enriched diet over 4 weeks. RESULTS: The highest number of genes differentially regulated by exercise in the liver was found in the mitogen-activated protein kinase (MAPK) signalling pathway, with a pronounced and transient upregulation of the transcription factors encoded by c-Fos (also known as Fos), c-Jun (also known as Jun), FosB (also known as Fosb) and JunB (also known as Junb) and phosphorylation of hepatic MAPK. Acute exercise also activated the p53 signalling pathway. A major role for oxidative stress is unlikely since the antioxidant-enriched diet did not prevent the activation of the MAPK pathway. In contrast, lower plasma glucose levels after running were related to enhanced levels of MAPK signalling proteins, similar to the upregulation of Igfbp1 and Pgc-1alpha (also known as Ppargc1a). In the working muscle the activation of the MAPK pathway was weak and not related to plasma glucose concentrations. CONCLUSIONS/INTERPRETATION: Metabolic stress evidenced as low plasma glucose levels appears to be an important determinant for the activation of the MAPK signalling pathway and the transcriptional response of the liver to acute exercise.

Genetic variations in angiogenesis pathway genes associated with clinical outcome in localized gastric adenocarcinoma.

BACKGROUND: Angiogenesis has been attributed to be a well-recognized aspect of human cancer biology. As such, proteinase-activated receptor (PAR)-1, endostatin (ES) and interleukin-8 (IL-8) mediate the regulation of early-onset angiogenesis and in turn impact the process of tumor-growth and disease progression. PATIENTS AND METHODS: Formalin-fixed paraffin-embedded tissues were obtained from 137 patients with localized gastric cancer at University of Southern California and Memorial Sloan-Kettering Cancer Center medical facilities. DNA was extracted and genotyping was carried out using PCR-restriction fragment length polymorphism-based protocols. RESULTS: In false discovery rate-adjusted univariate analysis, PAR-1 -506 ins/del (P < 0.001), ES +4349 G>A (P = 0.004), and IL-8 -251 T>A (P < 0.0001) were associated with time to tumor recurrence (TTR). Further, PAR-1 -506 ins/del and IL-8 -251 were associated with overall survival (OS). After adjusting for covariates, IL-8 remained significantly associated with TTR (adjusted P = 0.003) and OS (adjusted P = 0.049), whereas ES was significantly associated with TTR (adjusted P = 0.026). CONCLUSIONS: Polymorphisms in PAR-1, ES, and IL-8 may serve as independent molecular prognostic markers in patients with localized gastric adenocarcinoma. The assessment of the patients' individual risk on the basis of interindividual genotypes may therefore help to identify patient subgroups at high risk for poor clinical outcome.

The UCSC Genome Browser Database: 2008 update.

The University of California, Santa Cruz, Genome Browser Database (GBD) provides integrated sequence and annotation data for a large collection of vertebrate and model organism genomes. Seventeen new assemblies have been added to the database in the past year, for a total coverage of 19 vertebrate and 21 invertebrate species as of September 2007. For each assembly, the GBD contains a collection of annotation data aligned to the genomic sequence. Highlights of this year's additions include a 28-species human-based vertebrate conservation annotation, an enhanced UCSC Genes set, and more human variation, MGC, and ENCODE data. The database is optimized for fast interactive performance with a set of web-based tools that may be used to view, manipulate, filter and download the annotation data. New toolset features include the Genome Graphs tool for displaying genome-wide data sets, session saving and sharing, better custom track management, expanded Genome Browser configuration options and a Genome Browser wiki site. The downloadable GBD data, the companion Genome Browser toolset and links to documentation and related information can be found at: http://genome.ucsc.edu/.