Thesis (Ph.D.)--University of California, San Francisco, 2008. / Source: Dissertation Abstracts International, Volume: 69-10, Section: B, page: 5868. Adviser: David O. Morgan.

Thesis (Ph.D.)--University of California, San Francisco, 2010. / Source: Dissertation Abstracts International, Volume: 71-02, Section: B, page: . Adviser: Roland D. Mullins.

Thesis (Ph.D.)--University of California, San Francisco, 2010. / Source: Dissertation Abstracts International, Volume: 71-05, Section: B, page: . Advisers: Valerie M. Weaver; Nancy Boudreau.

Thesis (Ph.D.)--Indiana University, Dept. of Biochemistry, 2009. / Title from PDF t.p. (viewed on Jul 19, 2010). Source: Dissertation Abstracts International, Volume: 70-12, Section: B, page: 7308. Adviser: Claire E. Walczak.

Thesis (Ph.D.)--University of California, San Francisco, 2009. / Source: Dissertation Abstracts International, Volume: 70-10, Section: B, page: 5933. Adviser: Wallace F. Marshall.

Thesis (M.S.)--University of California, San Francisco, 2009. / Source: Masters Abstracts International, Volume: 48-03, page: 1476. Adviser: Linda C. Giudice.

<p> The identification of beige fat within the last decade, its ability to burn energy in adult humans, and its great potential for therapeutic applications has motivated many to work towards understanding how beige fat is made. Although several proteins have been identified as important for beige fat differentiation, it is clear that the differences that distinguish beige fat from other types of fat cannot be explained by the presence of these proteins alone. Additional regulatory proteins, including transcription factors and their co-factor proteins, must be involved. I took advantage of two important developments in the field of fat differentiation to develop two high throughput approaches that identified new transcription factors involved in beige fat: (1) our ability to culture beige fat cells by differentiating white fat pre-adipocytes in the presence of Rosiglitazone and (2) the established role for PRDM16 as required for beige fat differentiation. In brief, I combined RNA-seq data from beige fat cells and proteomics data from Rosiglitazone-dependent PRDM16 protein complexes to identify a set of candidate transcription factors involved in beige fat differentiation. The most promising candidate among this pool of putative beige fat regulatory transcription factors was GTF2IRD1. I determined that GTF2IRD1 is a PRDM16-interacting transcription factor that is enriched in beige and brown fat cells. In vivo, GTF2IRD1 is enriched in brown adipose tissue and is increased in beige and brown fat in response to beta-3-adrenergic stimulus. In the presence of the potent PPARgamma agonist Rosiglitazone, GTF2IRD1 overexpression enhances and shRNA-mediated knockdown reduces beige fat differentiation. GTF2IRD1 represses TGF-beta-mediated inhibition of beige fat differentiation. In summary, my data strongly supports that GTF2IRD1 is an essential regulator of beige fat differentiation through interaction with PRDM16 and inhibition of TGF-beta-mediated repression of differentiation.</p>

<p>The effects of high-voltage nanosecond electric pulses (nsEPs) on metastatic melanoma are still unclear. Hence, we applied one, two, three, and four 300 ns 40 kV/cm pulses to murine B16-F10 melanoma cells. Cell attachment ability was determined by comparing the number of floating cells and the percentage of attached cells. Melanoma inhibitory activity (MIA) is a secretory protein that is highly correlated with the malignancy and metastasis of malignant melanomas. We used MIA as our target to evaluate the effect of nsEPs on metastasis. Pulsed (experimental) and unpulsed (control) cells were incubated at 37&deg;C under a 5% CO₂ atmosphere. To determine cell attachment ability, the culture medium supernatant and attached cells were collected at 6, 12, 18, and 24 h after a single pulse. The live, dead, and total floating cells in the culture medium supernatant were counted. In addition, the live, dead, and total attached cells were counted after multiple pulses. Total RNA was extracted from the attached cells and reverse transcribed into cDNA. The MIA mRNA expression levels were measured using the cDNA temple via quantitative real-time PCR, with &beta;&ndash;actin as the internal control. The experiment was repeated three times (n=3). The results show that a single pulse did not affect the cell attachment ability, cell morphology, and the MIA mRNA expression levels (P=0.8058). Two pulses significantly decreased the cell attachment ability (P=0.014), cell viability (P&lt;0.0001), and changed the cell morphology, but did not change the MIA mRNA expression. The three-pulse and the four-pulse treatments significantly decreased the cell attachment ability (P=0.004, 0.00002, respectively), cell viability (P&lt;0.0001), changed the cell morphology, and increased the MIA mRNA expression levels within the first 12 h (P=0.041, 0.001, respectively). These indices were almost normal at 24 h after pulsing. We speculate that the two-, three-, and four-pulse treatments would be optimal for treating melanoma metastasis, whereas the single pulse treatment was not. Therefore, nsEPs provides a great opportunity for treating metastatic melanomas. </p>

Thesis (Ph.D.)--University of California, San Francisco, 2009. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3259. Adviser: Peter Walter.

<p> <b>Statement:</b> A global <i>Rbp1</i> knock out (<i> Rbp1-/-</i>) mouse model was used to correlate direct retinoid measurements with vitamin A metabolizing and atRA biosynthesizing enzyme activities, Crbp function and tissue microenvironment for the first time. </p><p> <b>Methods:</b> atRA was quantified by LC-MRM³ and ROL/RE/RAL was quantified by HPLC-UV. Enzyme activities were measured from enzymes present in subcellular fractions isolated from WT and <i>Rbp1-/- </i> tissues. Mouse CrbpI and CrbpIII were purified from transformed <i> Escherichia coli</i> for functional comparative studies. Tissue were formalin fixed for histological examination. Relative gene expression was analyzed using quantitative PCR. </p><p> <b>Results:</b> Reduced atRA was consistently quantified in extrahepatic tissues with elevated ROL/RE. Relative gene expression showed altered expression in retinoid pathway proteins and atRA loss preceded expression changes in some cases. Tissue microenvironments also consistently showed a loss of structure and organization along with accumulation of extracellular matrix and hyperplasia without apparent disease. Functional studies showed that CrbpIII binds retinol with less affinity than CrbpI and does not function equivalently to CrbpI in regulation of atRA biosynthesis. Also, metabolizing enzymes had altered activities in the <i>Rbp1-/-</i> tissues with reduced atRA biosynthesis. </p><p> <b>Conclusions:</b> Loss of CrbpI results in altered regulation of enzyme activity and atRA homeostasis cannot be maintained by other Crbp homologs in extrahepatic tissues. Dysfunctional atRA biosynthesis due to loss of CrbpI results in altered tissue microenvironment characteristic of dietary vitamin A deficiency and precancerous dysfunction associated with cancers that are observed to have silenced CrbpI.</p>