Changes in gene expression during the development of mammary tumors in MMTV-Wnt-1 transgenic mice

BACKGROUND: In human breast cancer normal mammary cells typically develop into hyperplasia, ductal carcinoma in situ, invasive cancer, and metastasis. The changes in gene expression associated with this stepwise progression are unclear. Mice transgenic for mouse mammary tumor virus (MMTV)-Wnt-1 exhibit discrete steps of mammary tumorigenesis, including hyperplasia, invasive ductal carcinoma, and distant metastasis. These mice might therefore be useful models for discovering changes in gene expression during cancer development. RESULTS: We used cDNA microarrays to determine the expression profiles of five normal mammary glands, seven hyperplastic mammary glands and 23 mammary tumors from MMTV-Wnt-1 transgenic mice, and 12 mammary tumors from MMTV-Neu transgenic mice. Adipose tissues were used to control for fat cells in the vicinity of the mammary glands. In these analyses, we found that the progression of normal virgin mammary glands to hyperplastic tissues and to mammary tumors is accompanied by differences in the expression of several hundred genes at each step. Some of these differences appear to be unique to the effects of Wnt signaling; others seem to be common to tumors induced by both Neu and Wnt-1 oncogenes. CONCLUSION: We described gene-expression patterns associated with breast-cancer development in mice, and identified genes that may be significant targets for oncogenic events. The expression data developed provide a resource for illuminating the molecular mechanisms involved in breast cancer development, especially through the identification of genes that are critical in cancer initiation and progression

Disruption, not displacement: Environmental variability and temporary migration in Bangladesh

Mass migration is one of the most concerning potential outcomes of global climate change. Recent research into environmentally induced migration suggests that relationship is much more complicated than originally posited by the ‘environmental refugee’ hypothesis. Climate change is likely to increase migration in some cases and reduce it in others, and these movements will more often be temporary and short term than permanent and long term. However, few large-sample studies have examined the evolution of temporary migration under changing environmental conditions. To address this gap, we measure the extent to which temperature, precipitation, and flooding can predict temporary migration in Matlab, Bangladesh. Our analysis incorporates high-frequency demographic surveillance data, a discrete time event history approach, and a range of sociodemographic and contextual controls. This approach reveals that migration declines immediately after flooding but quickly returns to normal. In contrast, optimal precipitation and high temperatures have sustained positive effects on temporary migration that persist over one to two year periods. Building on previous studies of long-term migration, these results challenge the common assumption that flooding, precipitation extremes and high temperatures will consistently increase temporary migration. Instead, our results are consistent with a livelihoods interpretation of environmental migration in which households draw on a range of strategies to cope with environmental variability

Decentralized Wastewater Management Solutions for Improved Public Health Protection and Reclamation: Optimization and Application

Centralized wastewater treatment, widely practiced in developed areas, involves transporting wastewater from large urban or industrial areas to a large capacity plant using a single network of sewers. Alternatively, the concept of wastewater collection, treatment and reuse at or near its point of generation is called decentralized wastewater treatment. Smaller decentralized plants with energy-efficient reclaimed water pumping, modularization of expansion and minimum capital investment can meet the increasing need for reclamation and wastewater management accessibility in rapidly developing regions Decentralized treatment can improve access to wastewater infrastructure in developing regions and improve energy-efficiency in reclamation in many rapidly growing developed regions. They can also replace land-intensive and inefficient treatment systems such as septic tanks and leach fields for remote, residential communities. They can reduce the strain on existing central facilities and can be an alternative to expensive collection system upgrades. We demonstrated the applicability of decentralized treatment to two examples in India and an example in Los Angeles. It is important to study the optimization and implications of decentralization. We formulated design and optimization methodology subject to user-defined constraints for a decentralized configuration of several treatment plants and collection networks. In this dissertation we developed an algorithm, using a constrained multi-objective optimization method (Genetic Algorithm), to obtain feasible decentralized configurations with minimum cost and energy. The applications of this methodology include obtaining the optimal solution for a given scenario, comparing alternative solutions devised by the user, and analyzing capacity expansion. Enabling easier reclamation and a more widespread access to wastewater collection and treatment are the chief motivation behind this study

Isolation of transforming growth factor-beta 2 cDNA from a fish, Cyprinus carpio by RT-PCR

Transforming Growth Factors-beta (TGF-beta s) have been described in many vertebrate species of amphibians, aves and mammals. In this report we demonstrate the presence of TGF-beta 2 in pisces. TGF-beta 2 has been cloned from a fish, Cyrinus carpio, by RT-PCR using degenerate oligonucleotide primers. Sequence analysis of the amplified product and alignment of the deduced amino acid sequence with the human TGF-beta 2 amino acid sequence revealed 81% and 93% identity in the precursor and the mature regions, respectively. The northern blot analysis of fish heart RNA shows a major messenger RNA species of about 8.0 kb and two messages of very low abundance of about 5.0 kb and 4.0 kb. The identification of TGF-beta 2 isoform in Pisces and it's high degree of homology with the mammalian isoform suggests that among all TGF-beta isoforms, TGF-beta 2 is the most conserved during evolution. (C) 1997 Elsevier Science B.V

Image_1_JMJD6 orchestrates a transcriptional program in favor of endocrine resistance in ER+ breast cancer cells.tif

High expression of Jumonji domain containing protein 6 (JMJD6) is strongly associated with poor prognosis in estrogen receptor positive (ER+) breast cancer. We overexpressed JMJD6 in MCF7 cells (JOE cells) and performed RNA-seq analysis. 76% of differentially expressed genes (DEGs) overlapped with ER target genes. Pathway analysis revealed that JMJD6 upregulated a larger subset of genes related to cell proliferation as compared to ER. Interestingly, JOE cells showed a decrease in ER target gene expression prompting us to check ER levels. Indeed, JOE cells showed a significant decrease in both ESR1 and ER levels and JMJD6 siRNA transfection increased the expression of both. Additionally, JOE cells showed increased RET and ERK1 expression, events associated with resistance to endocrine therapy. Accordingly, JOE cells displayed lower sensitivity and survived better at higher doses of 4-hydroxy tamoxifen (Tam) as compared to parental MCF-7 cells. Conversely, LTED-I and TAM R that resist Tam induced death, showed high expression of JMJD6. Further, JMJD6 siRNA treatment decreased growth and improved Tam sensitivity in TAM R. Comparison of JOE DEGs with known Tam signature genes showed a substantial overlap. Overall, these data suggest that blocking ER alone in patients may not eradicate proliferation of JMJD6 expressing ER+ cells and JMJD6 may predispose and sustain endocrine therapy resistance. We propose that immunostaining for JMJD6 could be developed as a potential marker for predicting endocrine therapy resistance. Further, antagonizing JMJD6 action in women expressing higher amounts of this protein, may offer a greater clinical benefit than endocrine therapy.</p

Table_3_JMJD6 orchestrates a transcriptional program in favor of endocrine resistance in ER+ breast cancer cells.xlsx

High expression of Jumonji domain containing protein 6 (JMJD6) is strongly associated with poor prognosis in estrogen receptor positive (ER+) breast cancer. We overexpressed JMJD6 in MCF7 cells (JOE cells) and performed RNA-seq analysis. 76% of differentially expressed genes (DEGs) overlapped with ER target genes. Pathway analysis revealed that JMJD6 upregulated a larger subset of genes related to cell proliferation as compared to ER. Interestingly, JOE cells showed a decrease in ER target gene expression prompting us to check ER levels. Indeed, JOE cells showed a significant decrease in both ESR1 and ER levels and JMJD6 siRNA transfection increased the expression of both. Additionally, JOE cells showed increased RET and ERK1 expression, events associated with resistance to endocrine therapy. Accordingly, JOE cells displayed lower sensitivity and survived better at higher doses of 4-hydroxy tamoxifen (Tam) as compared to parental MCF-7 cells. Conversely, LTED-I and TAM R that resist Tam induced death, showed high expression of JMJD6. Further, JMJD6 siRNA treatment decreased growth and improved Tam sensitivity in TAM R. Comparison of JOE DEGs with known Tam signature genes showed a substantial overlap. Overall, these data suggest that blocking ER alone in patients may not eradicate proliferation of JMJD6 expressing ER+ cells and JMJD6 may predispose and sustain endocrine therapy resistance. We propose that immunostaining for JMJD6 could be developed as a potential marker for predicting endocrine therapy resistance. Further, antagonizing JMJD6 action in women expressing higher amounts of this protein, may offer a greater clinical benefit than endocrine therapy.</p

Expression of transforming growth factor-beta isoforms in the rat male accessory sex organs and epididymis

We studied the expression and distribution of transforming growth factor-beta (TGF-beta) isoforms in the rat male accessory sex glands and the epididymis. Our data demonstrate the expression of both TGF-beta 1 and -beta 3 isoforms in ventral prostate (VP), seminal vesicle (SV), coagulating gland (CG), and epididymis (E) by Northern blot analysis. In addition, there was differential expression of TGF-beta 3 in the three regions of epididymis, the corpus region being the highest. Immunostaining data showed intense staining for latent TGF-beta 1 in all the male accessory glands. In contrast, no staining using antibodies specific for active TGF-beta 1 was observed. No expression of TGF-beta 2 was evident either by immunohistochemistry or Northern blot analysis. The presence of mature TGF-beta 3 protein was observed in the secretory epithelium of VP, CG, and corpus E. There was no detectable staining of TGF-beta 3 in the seminal vesicle and caput and cauda regions of epididymis. These data suggest possible differential regulation of TGF-beta isoform expression in the male reproductive system and predict unique roles for individual TGF-beta isoforms in sperm maturation and maintenance

Image_3_JMJD6 orchestrates a transcriptional program in favor of endocrine resistance in ER+ breast cancer cells.tif

High expression of Jumonji domain containing protein 6 (JMJD6) is strongly associated with poor prognosis in estrogen receptor positive (ER+) breast cancer. We overexpressed JMJD6 in MCF7 cells (JOE cells) and performed RNA-seq analysis. 76% of differentially expressed genes (DEGs) overlapped with ER target genes. Pathway analysis revealed that JMJD6 upregulated a larger subset of genes related to cell proliferation as compared to ER. Interestingly, JOE cells showed a decrease in ER target gene expression prompting us to check ER levels. Indeed, JOE cells showed a significant decrease in both ESR1 and ER levels and JMJD6 siRNA transfection increased the expression of both. Additionally, JOE cells showed increased RET and ERK1 expression, events associated with resistance to endocrine therapy. Accordingly, JOE cells displayed lower sensitivity and survived better at higher doses of 4-hydroxy tamoxifen (Tam) as compared to parental MCF-7 cells. Conversely, LTED-I and TAM R that resist Tam induced death, showed high expression of JMJD6. Further, JMJD6 siRNA treatment decreased growth and improved Tam sensitivity in TAM R. Comparison of JOE DEGs with known Tam signature genes showed a substantial overlap. Overall, these data suggest that blocking ER alone in patients may not eradicate proliferation of JMJD6 expressing ER+ cells and JMJD6 may predispose and sustain endocrine therapy resistance. We propose that immunostaining for JMJD6 could be developed as a potential marker for predicting endocrine therapy resistance. Further, antagonizing JMJD6 action in women expressing higher amounts of this protein, may offer a greater clinical benefit than endocrine therapy.</p

Comparative Functional Analysis of Rat TGF−β1TGF-\beta 1 and Xenopus laevis TGF−β1TGF-\beta 1 Promoters Suggest Differential Regulations

We have carried out a comparative functional analysis of the rat $TGF-\beta 1$ and Xenopus laevis $TGF-\beta 5$ promoters across several mammalian and amphibian cell lines. Progressive deletion constructs of both the promoters have been made using a PCR based approach and the basal promoter activities studied in Xenopus tadpole cell line (XTC), Xenopus adult kidney fibroblast cell line (A6), human hepatoma cell line (HepG2), normal rat kidney cell line (NRK), and Chinese hamster ovary cell line (CHO). Data suggests that the basal promoter activity of $TGF-\beta 1$ is low as compared to $TGF-\beta 5$ promoter in XTC cells but comparable in A6 cells, while $TGF-\beta 5$ promoter shows nearly negligible activity as compared to TGF-beta5 promoter in all the tested mammalian cell lines. Moreover, $TGF-\beta 5$ promoter is found to be repressed in XTC cells on treatment with $TGF-\beta 5$ protein. Thus, the regulation of $TGF-\beta 1$ and $TGF-\beta 5$ promoters is distinct in amphibian and mammalian species. We therefore suggest that contrary to the suggested functional equivalence of $TGF-\beta 1$ and $TGF-\beta 5$ proteins, $TGF-\beta 1$ and $TGF-\beta 5$ genes have distinct functions in their respective species