Correction: Genistein downregulates onco-miR-1260b and upregulates sFRP1 and Smad4 via demethylation and histone modification in prostate cancer cells.

The authors report that there is a mistake in the representative picture of Fig. 4D (top row: PC3-miR1260b inh-0h) in the original version. The correct version of Fig. 4 with the original pictures for both PC3 miR-NC inh-0h and PC3-miR1260b inh-0h are provided below.

Climate change impact of livestock CH4 emission in India: Global temperature change potential (GTP) and surface temperature response.

Two climate metrics, Global surface Temperature Change Potential (GTP) and the Absolute GTP (AGTP) are used for studying the global surface temperature impact of CH4 emission from livestock in India. The impact on global surface temperature is estimated for 20 and 100 year time frames due to CH4 emission. The results show that the CH4 emission from livestock, worked out to 15.3 Tg in 2012. In terms of climate metrics GTP of livestock-related CH4 emission in India in 2012 were 1030 Tg CO2e (GTP20) and 62 Tg CO2e (GTP100) at the 20 and 100 year time horizon, respectively. The study also illustrates that livestock-related CH4 emissions in India can cause a surface temperature increase of up to 0.7mK and 0.036mK over the 20 and 100 year time periods, respectively. The surface temperature response to a year of Indian livestock emission peaks at 0.9mK in the year 2021 (9 years after the time of emission). The AGTP gives important information in terms of temperature change due to annual CH4 emissions, which is useful when comparing policies that address multiple gases.

Genistein downregulates onco-miR-1260b and upregulates sFRP1 and Smad4 via demethylation and histone modification in prostate cancer cells.

BACKGROUND: Recently several microRNAs (miRNAs) have been found to be regulated by genistein in cancer cells. In this study, we focused on the gene regulatory effect of genistein on microRNA and its target genes in prostate cancer (PC). METHODS: Initially, we investigated the effect of genistein on prostate cancer cells and identified that the expression of miRNA-1260b was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA-1260b expression and prostate cancer patient outcomes. Two target genes (sFRP1 and Smad4) of miR-1260b were identified based on computer algorithm and 3'UTR luciferase assay was carried out to determine direct miRNA regulation of the genes. RESULTS: Genistein promoted apoptosis while inhibiting prostate cancer cell proliferation, invasion and TCF reporter activity in PC cells. MiR-1260b was highly expressed in prostate cancer tissues and significantly downregulated by genistein in PC cells. After knocking down miR-1260b, cell proliferation, invasion, migration and TCF reporter activity were decreased in PC cells. Western analysis and 3'UTR luciferase assay showed that the two target genes (sFRP1 and Smad4) were directly regulated by miR-1260b. The expression of sFRP1 and Smad4 was significantly decreased in prostate cancer tissues. Genistein also increased expression of these two genes via DNA demethylation and histone modifications. CONCLUSIONS: Our data suggest that genistein exerts its anti-tumour effect via downregulation of miR-1260b that targeted sRRP1 and Smad4 genes in prostate cancer cells. The expression of sFRP1 and Smad4 was also modulated by genistein via DNA methylation or histone modifications in PC cell lines.

microRNA-183 is an oncogene targeting Dkk-3 and SMAD4 in prostate cancer.

BACKGROUND: The purpose of this study was to identify prostate cancer (PC) oncogenic microRNAs (miRs) based on miR microarray and to investigate whether these oncogenic miRs may be useful as PC biomarkers. METHODS: Initially, we carried out miR microarray and real-time PCR using RWPE-1, PC-3, DU-145 and LNCaP cells. To investigate the function of miR-183, we used a miR-183 knockdown inhibitor in cell growth and wound-healing assays. We used several algorithms and confirmed that they are directly regulated by miR-183. RESULTS: We identified three potential oncogenic miRs (miR-146a, miR-183 and miR-767-5P). The expression of miR-183 in PC cells (PC-3, DU-145 and LNCaP) was upregulated compared with RWPE-1 cells. MiR-183 expression was also significantly higher in PC tissues compared with that in matched normal prostate tissues. Additionally, miR-183 expression was correlated with higher prostate-specific antigen, higher pT and shorter overall survival. MiR-183 knockdown decreased cell growth and motility in PC cells and significantly decreased prostate tumour growth in in vivo nude mice experiments. We identified Dkk-3 and SMAD4 as potential target genes of miR-183. CONCLUSION: Our data suggest that oncogenic miR-183 may be useful as a new PC biomarker and that inhibition of miR-183 expression may be therapeutically beneficial as a PC treatment.

Genistein downregulates onco-miR-1260b and inhibits Wnt-signalling in renal cancer cells.

BACKGROUND: Wnt-signalling has an important role in renal cancer and it is modulated by genistein in other cancers. Recently, microRNAs (miRNAs) have emerged as new regulators of gene expression. Thus, we focused on miRNAs to examine the regulatory mechanism of genistein on the Wnt-signalling pathway in renal cell carcinoma (RCC). METHODS: Initially, we investigated the effect of genistein on Wnt-signalling (TOPflash reporter assay (TCF reporter assays)) in renal cancer cells, and using microarray identified candidate miRNAs whose expression was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA expression and renal cancer patient outcomes. We also did 3'UTR luciferase assays to look at direct miRNA regulation of Wnt-signalling-related genes. RESULTS: Genistein promoted apoptosis while inhibiting RCC cell proliferation and invasion. Genistein also decreased TCF reporter activity in RCC cells. We found that miR-1260b was highly expressed and significantly downregulated by genistein in RCC cells. The expression of miR-1260b was significantly higher in renal cancer tissues compared with normal, and significantly related to overall shorter survival. In addition, miR-1260b promoted renal cancer cell proliferation and invasion in RCC cells. The 3'UTR luciferase activity of target genes (sFRP1, Dkk2, Smad4) was significantly decreased and their protein expression significantly upregulated in miR-1260b inhibitor-transfected renal cancer cells. CONCLUSION: Our data suggest that genistein inhibited Wnt-signalling by regulating miR-1260b expression in renal cancer cells.

Tumour suppressor microRNA-584 directly targets oncogene Rock-1 and decreases invasion ability in human clear cell renal cell carcinoma.

BACKGROUND: The purpose of this study was to identify new tumour suppressor microRNAs (miRs) in clear cell renal cell carcinoma (ccRCC), carry out functional analysis of their suppressive role and identify their specific target genes. METHODS: To explore suppressor miRs in RCC, miR microarray and real-time PCR were performed using HK-2 and A-498 cells. Cell viability, invasion and wound healing assays were carried out for functional analysis after miR transfection. To determine target genes of miR, we used messenger RNA (mRNA) microarray and target scan algorithms to identify target oncogenes. A 3'UTR luciferase assay was also performed. Protein expression of target genes in ccRCC tissues was confirmed by immunohistochemistry and was compared with miR-584 expression in ccRCC tissues. RESULTS: Expression of miR-584 in RCC (A-498 and 769-P) cells was downregulated compared with HK-2 cells. Transfection of miR-584 dramatically decreased cell motility. The ROCK-1 mRNA was inhibited by miR-584 and predicted to be target gene. The miR-584 decreased 3'UTR luciferase activity of ROCK-1 and ROCK-1 protein expression. Low expression of miR-584 in ccRCC tissues was correlated with high expression of ROCK-1 protein. The knockdown of ROCK-1 by siRNA inhibited cell motility. CONCLUSION: miR-584 is a new tumour suppressor miR in ccRCC and inhibits cell motility through downregulation of ROCK-1.

The functional significance of microRNA-145 in prostate cancer.

BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that have important roles in numerous cellular processes. Recent studies have shown aberrant expression of miRNAs in prostate cancer tissues and cell lines. On the basis of miRNA microarray data, we found that miR-145 is significantly downregulated in prostate cancer. METHODS AND RESULTS: We investigated the expression and functional significance of miR-145 in prostate cancer. The expression of miR-145 was low in all the prostate cell lines tested (PC3, LNCaP and DU145) compared with the normal cell line, PWR-1E, and in cancerous regions of human prostate tissue when compared with the matched adjacent normal. Overexpression of miR-145 in PC3-transfected cells resulted in increased apoptosis and an increase in cells in the G2/M phase, as detected by flow cytometry. Investigation of the mechanisms of inactivation of miR-145 through epigenetic pathways revealed significant DNA methylation of the miR-145 promoter region in prostate cancer cell lines. Microarray analyses of miR-145-overexpressing PC3 cells showed upregulation of the pro-apoptotic gene TNFSF10, which was confirmed by real-time PCR and western analysis. CONCLUSION: One of the genes significantly upregulated by miR-145 overexpression is the proapoptotic gene TNFSF10. Therefore, modulation of miR-145 may be an important therapeutic approach for the management of prostate cancer.

Methane production and estimation from livestock husbandry: A mechanistic understanding and emerging mitigation options.

Globally, livestock is an important contributor to methane (CH4) emissions. This paper reviewed the various CH4 measurement and estimation techniques and mitigation approaches for the livestock sector. Two approaches for enteric livestock CH4 emission estimation are the top-down and bottom-up. The combination of both could further improve our understanding of enteric CH4 emission and possible mitigation measures. We discuss three mitigation approaches: reducing emissions, avoiding emissions, and enhancing the removal of emissions from livestock. Dietary management, livestock management, and breeding management are viable reducing emissions pathways. Dietary manipulation is easily applicable and can bring an immediate response. Economic incentive policies can help the livestock farmers to opt for diet, breeding, and livestock management mitigation approaches. Carbon pricing creates a better option to achieve reduction targets in a given period. A combination of carbon pricing, feeding management, breeding management, and livestock management is more feasible and sustainable CH4 emissions mitigation strategy rather than a single approach.

Down-regulation of frizzled-7 expression decreases survival, invasion and metastatic capabilities of colon cancer cells.

BACKGROUND: The canonical Wnt signalling pathway is activated in most sporadic colorectal cancers (CRCs). We previously reported that FZD7 functions as a receptor for the canonical Wnt signalling pathway in colon cancer cells. METHODS AND RESULTS: In this study, we examined the function of FZD7 in survival, invasion and metastatic capabilities of colon cancer cells. FZD7_siRNA transfection decreased cell viability of HT-29 and HCT-116 colon cancer cells. Expression of c-Jun, phosphorylation of JNK and c-Jun, and activation of RhoA were suppressed after FZD7_siRNA transfection into HCT-116 cells. In vitro invasion activity and Wnt target gene expression were also reduced in HCT-116 cells transfected with FZD7_siRNA. Liver metastasis of stable FZD7_siRNA HCT-116 cell transfectants in scid mice was decreased to 40-50% compared to controls. The mRNA levels of FZD7 in 135 primary CRC tissues were examined by real-time PCR. FZD7 mRNA levels were significantly higher in stage II, III or IV tumours than in non-tumour tissues (P<0.005), and overall survival was shorter in those patients with higher FZD7 expression (P<0.001). CONCLUSION: These data suggest that FZD7 may be involved in enhancement of survival, invasion and metastatic capabilities of colon cancer cells through non-canonical Wnt signalling pathways as well as the canonical pathway.

Propagation of amorphous oxide nanowires via the VLS mechanism: growth kinetics.

This work reports the growth kinetics of amorphous nanowires (NWs) developed by the vapour-liquid-solid (VLS) mechanism. The model presented here incorporates all atomistic processes contributing to the growth of amorphous oxide NWs having diameters in the 5-100 nm range. The steady state growth condition has been described by balancing the key atomistic process steps. It is found that the 2D nano-catalyst liquid and NW solid (L-S) interface plays a central role in the kinetic analysis. The balance between the 2D Si layer crystallization and oxidation rate is quantitatively examined and compared with experimental values. The atomistic process dependencies of the NW growth rate, supersaturation (C/C 0), desolvation energy (Q D) barrier and NW diameter have been analyzed in detail. The model successfully predicts the reported NW growth rate to be in the range of 1-10 µm s-1. A novel seed/catalyst metal-based synthesis strategy for the preparation of amorphous silica NWs is reported. A nickel thin film on Si is used as a seed metal for the Au assisted VLS growth of silica NWs. The experimental results provide evidence of the creation of SiO under the given conditions followed by Si injection in the Au-Si nano-catalyst solution. The usage of seed metal was observed to reduce the growth temperature compared to the methods reported in the literature and obtain similar growth rates. The technique presented here holds promise for the synthesis of sub-100 nm diameter NWs.

COD and BOD removal from domestic wastewater generated in decentralised sectors.

The objective of this work is to study COD and BOD reduction of domestic wastewater using discarded material based mixed adsorbents (mixed adsorbent carbon, MAC and commercial activated carbon, CAC) in batch mode. Under optimum conditions, maximum reduction and maximum COD and BOD reduction achieved using MAC and CAC was 95.87% and 97.45%; and 99.05% and 99.54%, respectively. Results showed that MAC offered potential benefits for COD and BOD removal from wastewater.

State of municipal solid waste management in Delhi, the capital of India.

Delhi is the most densely populated and urbanized city of India. The annual growth rate in population during the last decade (1991-2001) was 3.85%, almost double the national average. Delhi is also a commercial hub, providing employment opportunities and accelerating the pace of urbanization, resulting in a corresponding increase in municipal solid waste (MSW) generation. Presently the inhabitants of Delhi generate about 7000tonnes/day of MSW, which is projected to rise to 17,000-25,000tonnes/day by the year 2021. MSW management has remained one of the most neglected areas of the municipal system in Delhi. About 70-80% of generated MSW is collected and the rest remains unattended on streets or in small open dumps. Only 9% of the collected MSW is treated through composting, the only treatment option, and rest is disposed in uncontrolled open landfills at the outskirts of the city. The existing composting plants are unable to operate to their intended treatment capacity due to several operational problems. Therefore, along with residue from the composting process, the majority of MSW is disposed in landfills. In absence of leachate and landfill gas collection systems, these landfills are a major source of groundwater contamination and air pollution (including generation of greenhouse gases). This study describes and evaluates the present state of municipal solid waste management in Delhi. The paper also summarizes the proposed policies and initiatives of the Government of Delhi and the Municipal Corporation of Delhi to improve the existing MSW management system.

Premalignant alterations in the lipid composition and fluidity of colonic brush border membranes of rats administered 1,2 dimethylhydrazine.

Dimethylhydrazine (DMH) is a potent procarcinogen with selectivity for the colon. To determine whether alterations in the lipid composition and fluidity of rat colonic brush border membranes existed before the development of DMH-induced colon cancer, rats were injected s.c. with this agent (20 mg/kg body weight per wk) or diluent for 5, 10, and 15 wk. Animals were killed at these time periods and brush border membranes were prepared from proximal and distal colonocytes of each group. The "static" and "dynamic" components of fluidity of each membrane were then assessed, by steady-state fluorescence polarization techniques using limiting hindered fluorescence anisotropy and order parameter values of the fluorophore 1,6 diphenyl-1,3,5-hexatriene (DPH) and fluorescence anisotropy values of DL-2-(9-anthroyl) stearic acid and DL-12-(9-anthroyl) stearic acid, respectively. Membrane lipids were extracted and analyzed by thin-layer chromatography and gas-liquid chromatography. Phospholipid methylation activity in these membranes was also measured using S-adenosyl-L-methionine as the methyl donor. The results of these studies demonstrate that: the lipid composition and both components of fluidity of proximal DMH-treated and control membranes and their liposomes were similar at all time periods examined; at 5, 10, and 15 wk the "dynamic component of fluidity" of distal DMH-treated membranes and their liposomes was found to be higher, similar, and lower, respectively, than their control counterparts; the "static component of fluidity" of distal DMH-treated membranes and their liposomes, however, was similar to control preparations at all three time periods; and alterations in the lipid composition and phospholipid methylation activities appeared to be responsible for these differences in the "dynamic component of fluidity" at these various time periods.

Mucin production by human colonic carcinoma cells correlates with their metastatic potential in animal models of colon cancer metastasis.

Patients with mucinous colorectal cancers characteristically present with advanced disease, however, the relationship between mucin production by colon cancer cells and their metastatic potential remains unclear. We therefore sought to define the relationship between mucin production by human colon cancer cells and metastatic ability by employing animal models of colon cancer metastasis. LS LiM 6, a colon carcinoma cell line with high liver metastasizing ability during cecal growth in nude mice produced twofold more metabolically labeled intracellular mucin and secreted four- to fivefold more mucin into the culture medium compared to poorly metastatic parental line LS174T. This was accompanied by a similar elevation in poly(A)+ RNA detected by blot hybridization with a human intestinal mucin cDNA probe, and increases in mucin core carbohydrate antigens determined immunohistochemically. Variants of LS174T selected for high (HM 7) or low (LM 12) mucin synthesizing capacity also yielded metastases after cecal growth and colonized the liver after splenic-portal injection in proportion to their ability to produce mucin. Inhibition of mucin glycosylation by the arylglycoside benzyl-alpha-N-acetyl-galactosamine greatly reduced liver colonization after splenic-portal injection of the tumor cells. These data suggest that mucin production by human colon cancer cells correlates with their metastatic potential and affects their ability to colonize the liver in experimental model systems.

MicroRNA-383 located in frequently deleted chromosomal locus 8p22 regulates CD44 in prostate cancer.

A major genomic alteration in prostate cancer (PCa) is frequent loss of chromosome (chr) 8p with a common region of loss of heterozygosity (LOH) at chr8p22 locus. Genomic studies implicate this locus in the initiation of clinically significant PCa and with progression to metastatic disease. However, the genes within this region have not been fully characterized to date. Here we demonstrate for the first time that a microRNA component of this region-miR-383-is frequently downregulated in prostate cancer, has a critical role in determining tumor-initiating potential and is involved in prostate cancer metastasis via direct regulation of CD44, a ubiquitous marker of PCa tumor-initiating cells (TICs)/stem cells. Expression analyses of miR-383 in PCa clinical tissues established that low miR-383 expression is associated with poor prognosis. Functional data suggest that miR-383 regulates PCa tumor-initiating/stem-like cells via CD44 regulation. Ectopic expression of miR-383 inhibited tumor-initiating capacity of CD44+ PCa cells. Also, 'anti-metastatic' effects of ectopic miR-383 expression were observed in a PCa experimental metastasis model. In view of our results, we propose that frequent loss of miR-383 at chr8p22 region leads to tumor initiation and prostate cancer metastasis. Thus, we have identified a novel finding that associates a long observed genomic alteration to PCa stemness and metastasis. Our data suggest that restoration of miR-383 expression may be an effective therapeutic modality against PCa. Importantly, we identified miR-383 as a novel PCa tissue diagnostic biomarker with a potential that outperforms that of serum PSA.

Municipal solid waste characterization and its assessment for potential methane generation: a case study.

There has been a significant increase in municipal solid waste (MSW) generation in India during the last few decades and its management has become a major issue because the poor waste management practices affect the health and amenity of the cities. In the present study, various physico-chemical parameters of the MSW were analyzed to characterize the waste dumped at Gazipur landfill site in Delhi, India, which shows that it contains a high fraction of degradable organic components. The decomposition of organic components produces methane, a significant contributor to global warming. Based on the waste composition, waste age and the total amount dumped, a first-order decay model (FOD) was applied to estimate the methane generation potential of the Gazipur landfill site, which yields an estimate of 15.3 Gg/year. This value accounts to about 1-3% of existing Indian landfill methane emission estimates. Based on the investigation of Gazipur landfill, we estimate Indian landfill methane emissions at 1.25 Tg/year or 1.68 Tg/year of methane generation potential. These values are within the range of existing estimates. A comparison of FOD with a recently proposed triangular model was also performed and it shows that both models can be used for the estimation of methane generation. However, the decrease of the emission after closure is more gradual in the case of the first-order model, leading to larger gas production predictions after more than 10 years of closure. The regional and global implications of national landfill methane emission are also discussed.

Induction of enhanced methane oxidation in compost: temperature and moisture response.

Landfilling is one of the most common ways of municipal solid waste disposal. Degradation of organic waste produces CH(4) and other landfill gases that significantly contribute to global warming. However, before entering the atmosphere, part of the produced CH(4) can be oxidised while passing through the landfill cover. In the present study, the oxidation rate of CH(4) was studied with various types of compost as possible landfill cover. The influence of incubation time, moisture content and temperature on the CH(4) oxidation capacity of different types of compost was examined. It was observed that the influence of moisture content and temperature on methane oxidation is time-dependent. Maximum oxidation rates were observed at moisture contents ranging from 45% to 110% (dry weight basis), while the optimum temperature ranged from 15 to 30 degrees C.

Premalignant alterations in the glycosphingolipid composition of colonic epithelial cells of rats treated with 1,2-dimethylhydrazine.

1,2-Dimethylhydrazine is a procarcinogen with selectivity for the colon. In weekly s.c. doses of 20 mg/kg of body weight, this agent produces colonic tumors in virtually 100% of rodents, with a latency period of approximately 6 months. To determine whether alterations in the glycosphingolipid content and composition of rat colonic epithelial cells existed before the development of dimethylhydrazine-induced colon cancer, rats were given s.c. injections of this agent (20 mg/kg body weight per week) or diluent for 5 weeks. Animals were sacrificed at this time period and colonocytes isolated from each group. Glycosphingolipids were then extracted from these cells and analyzed by thin-layer chromatography, high-performance liquid chromatography and gas-liquid chromatography. The results of these studies demonstrate that: (a) the content and relative percentages of globotriaosylceramide is increased, whereas hematoside and globotetraosylceramide are decreased in dimethylhydrazine-treated colonocytes compared to their control counterparts; and (b) differences in the enzymatic activities responsible for the biosynthesis of these glycosphingolipids appear to explain, at least partially, these compositional differences. The present data, therefore, suggest that alterations in certain glycosphingolipids may be an early event in colonic malignant transformation and, furthermore, that these alterations may prove useful in the detection of early colon cancer.