UBE2L6/UBCH8 and ISG15 attenuate autophagy in esophageal cancer cells

Esophageal cancer remains a poor prognosis cancer due to advanced stage of presentation and drug resistant disease. To understand the molecular mechanisms influencing response to chemotherapy, we examined genes that are differentially expressed between drug sensitive, apoptosis competent esophageal cancer cells (OE21, OE33, FLO-1) and those which are more resistant and do not exhibit apoptosis (KYSE450 and OE19). Members of the ISG15 (ubiquitin-like) protein modification pathway, including UBE2L6 and ISG15, were found to be more highly expressed in the drug sensitive cell lines. In this study, we evaluated the contribution of these proteins to the response of drug sensitive cells. Depletion of UBE2L6 or ISG15 with siRNA did not influence caspase-3 activation or nuclear fragmentation following treatment with 5-fluorouracil (5-FU). We assessed autophagy by analysis of LC3II expression and Cyto-ID staining. Depletion of either ISG15 or UBE2L6 resulted in enhanced endogenous autophagic flux. An increase in autophagic flux was also observed following treatment with cytotoxic drugs (5-FU, rapamycin). In ISG15 depleted cells, this increase in autophagy was associated with improved recovery of drug treated cells. In contrast, UBE2L6 depleted cells, did not show enhanced recovery. UBE2L6 may therefore influence additional targets that limit the pro-survival effect of ISG15 depletion. These data identify UBE2L6 and ISG15 as novel inhibitors of autophagy, with the potential to influence chemosensitivity in esophageal cancer cells

Genotoxic and histotoxic effects of air pollutants at a benzene station on albino rats

Background: This study was designed to explore the hazardous effects of occupational exposure to air pollutants arising from benzene stations.Methods: A total of 48 albino rats were divided into three groups each of sixteen animals. Groups-I and II were kept at a benzene station for 60 and 120 days, respectively; while group-III was kept as a control under normal laboratory conditions. At the end of the experiment, animals were sacrificed and bone marrow samples were taken to investigate the effect of air pollutants at a benzene station on chromosomal aberrations and micronuclei formation. Tissue samples were also picked out to evaluate histotoxic effects of these air pollutants.Results: Results indicated pronounced time-dependent effects because of the work place pollutants. Genetic examination revealed higher frequency of chromosomal aberrations and elevated mitotic index after exposure to air pollutants. Micronuclei formation also increased either in polychromatic or normochromatic erythrocyte due to air pollutants. Histopathological examination revealed congestion of the pulmonary blood vessels with pulmonary edema. Bronchial hyperplasia and metaplasia were also seen in late stage of exposure. The liver showed degenerative changes or even necrosis of hepatocytes, while the kidneys showed congestion of the renal blood vessels and glomerular tufts. Hyalinization of the glomeruli was also detected. The heart showed various mononuclear infiltration and the testes showed degenerative changes of seminiferous tubules. Spleen showed hemosiderosis while testes showed degenerative changes of seminiferous tubules.Conclusions: The findings of the present study indicated that benzene exposure may lead to toxic effects including, genotoxicities and histotoxicities. In order to minimize the predicted toxic effect of occupational exposure to benzene the strict protective measures should be put in consideration

LC3B globular structures correlate with survival in esophageal adenocarcinoma

Background: Esophageal adenocarcinoma has the fastest growing incidence of any solid tumor in the Western world. Prognosis remains poor with overall five-year survival rates under 25 %. Only a limited number of patients benefit from chemotherapy and there are no biomarkers that can predict outcome. Previous studies have indicated that induction of autophagy can influence various aspects of tumor cell biology, including chemosensitivity. The objective of this study was to assess whether expression of the autophagy marker (LC3B) correlated with patient outcome. Methods: Esophageal adenocarcinoma tumor tissue from two independent sites, was examined retrospectively. Tumors from 104 neoadjuvant naïve patients and 48 patients post neoadjuvant therapy were assembled into tissue microarrays prior to immunohistochemical analysis. Kaplan-Meier survival curves and log-rank tests were used to assess impact of LC3B expression on survival. Cox regression was used to examine association with clinical risk factors. Results: A distinct globular pattern of LC3B expression was found to be predictive of outcome in both patient groups, irrespective of treatment (p < 0.001). Multivariate analysis found that this was a strong independent predictor of poor prognosis (p < 0.001). Conclusions: This distinctive staining pattern of LC3B represents a novel prognostic marker for resectable esophageal adenocarcinoma

Determination of mixing quality in biogas plant digesters using tracer tests and computational fluid dynamics

The total electricity demand of investigated biogas plants (BGP) makes up 7–8 % of the total electricity produced. Nearly 40 % of this energy is consumed just for mixing in digesters and the energy demand for mixing in some biogas plants can be even higher. Therefore, optimal mixing in anaerobic digesters is a basic condition for efficient plant operation and biogas production. The use of problematic substrates (e.g. grass silage or other fibrous substrates), installation of unsuitable mixing systems or inconvenient mixing intervals may lead to mixing problems. Knowledge about mixing in biogas digesters is still insufficient, so the objective of this study was to fill the information gaps in the literature by determining the minimal retention time of substrates fed into anaerobic digesters and to describe substrate distribution and washing out rates from investigated digesters. Two full-scale biogas plant digesters (2000 m3 and 1500 m3) using different mixing systems and substrates were investigated. To characterize the substrate distribution, lithium hydroxide monohydrate solutions were used for tracer tests at concentrations of 47.1 mg Li+ / kg TS and 46.6 mg Li+ / kg TS in digester. The tracer concentration in the digester effluents was measured during two hydraulic retention times and compared. Although the tracer was detected in the digester effluent at nearly the same time in both cases, the tracer tests showed very different distribution curves. The tracer concentration in effluent B grew much slower than in effluent A and no significant short circuiting streams were detected. Although the data calculated by computational fluid dynamics methods (CFD) showed a very good agreement with the full scale results, full comparison was not possible

Genotypic detection of rifampicin and isoniazid resistant Mycobacterium tuberculosis strains by DNA sequencing: a randomized trial

Correction to Genotypic detection of rifampicin and isoniazid resistant Mycobacterium tuberculosis strains by DNA sequencing: a randomized trial Amina Abdelaal, Hassan Abd El-Ghaffar, Mohammad Hosam Eldeen Zaghloul, Noha El mashad, Ehab Badran, Amal Fathy Annals of Clinical Microbiology and Antimicrobials 2009, 8:4 (30 January 2009

Fan and Ventilation Rate Monitoring of Cage-Free Layer Houses in California

Ventilation rates were continuously monitored in two cage-free layer houses located in California from March 1, 2012 to May 13, 2013. The average number of brown Lohmann laying hens in each house was 33,300. Temperature, relative humidity, static pressure, and running status of 48 ventilation fans were continuously monitored and recorded every minute. Regression models were developed to relate house temperature and ventilation rate to inlet air temperature, and to relate airflow rate to building static pressure (R2 = 0.98). Results showed that the daily mean ventilation rate per hen ranged from 1.91 to 8.72 m3 h-1 hen-1, averaging at 4.49 ±1.53 m3 h-1 hen-1. The standard uncertainty of daily mean ventilation rate was determined to be 3.7%. The 91-cm and 130-cm fans were found to perform at 82% and 63% of the manufacturer rated airflow rates, respectively. The minimum and maximum static pressure was 11.0 and 50.6 Pa, respectively, corresponding to 2 and 16 running tunnel fans. When the house temperature exceeded 30ºC, an evaporative cooling system was activated, which could reduce the inlet air temperature by 6.3ºC and concurrently increased the indoor air humidity ratio by 3.4 g per kg dry air. Cooling pad efficiency was 66%. The sidewall fans and tunnel fans were operated at 65% and 20% of the total time when layers occupied the houses. The new rational formula to calculate dry base ventilation rates was developed based upon the ratio of water vapor volume to moist air volume. The developed models and data collected in this research can be used to calculate the ventilation rates in cage-free layer houses so that it can be possible to assure healthy conditions needed for layers. They can also be used in the design of cage free houses and in the calculations of emissions of air pollutants from these houses