Mammalian SWI/SNF Enzymes and the Epigenetics of Tumor Cell Metabolic Reprogramming

Tumor cells reprogram their metabolism to survive and grow in a challenging microenvironment. Some of this reprogramming is performed by epigenetic mechanisms. Epigenetics is in turn affected by metabolism; chromatin modifying enzymes are dependent on substrates that are also key metabolic intermediates. We have shown that the chromatin remodeling enzyme Brahma-related gene 1 (BRG1), an epigenetic regulator, is necessary for rapid breast cancer cell proliferation. The mechanism for this requirement is the BRG1-dependent transcription of key lipogenic enzymes and regulators. Reduction in lipid synthesis lowers proliferation rates, which can be restored by palmitate supplementation. This work has established BRG1 as an attractive target for breast cancer therapy. Unlike genetic alterations, epigenetic mechanisms are reversible, promising gentler therapies without permanent off-target effects at distant sites

BRG1, a SWI/SNF chromatin remodeling enzyme ATPase, is required for maintenance of nuclear shape and integrity

We recently reported that reducing the levels of BRG1, the catalytic subunit of mammalian SWI/SNF chromatin remodeling enzymes, induces alterations in nuclear shape in a breast epithelial cell line. Immunostaining the BRG1 knockdown cells with nuclear lamina antibodies revealed a significantly increased frequency of grooves, or invaginations, in the nuclei. Disruption of each of the major cytoplasmic filament systems (actin, tubulin and cytokeratins) had no impact on the BRG1-dependent changes in nuclear shape, indicating that the observed changes in nuclear morphology are unlikely to be a result of alterations in the integrity of the nuclear-cytoplamic contacts in the cell. We propose that the BRG1-dependent nuclear shape changes reflect a role for the chromatin remodeling enzyme in maintaining the structural integrity of the nucleus via global regulation of chromatin structure and dynamics within the nucleus

Impact of maximum borehole depths on inverted temperature histories in borehole paleoclimatology

A quantitative assessment is presented for the impact of the maximum depth of a temperature-depth profile on the estimate of the climatic transient and the resultant ground surface temperature (GST) reconstruction used in borehole paleoclimatology. The depth of the profile is important because the downwelling climatic signal must be separated from the quasi-steady state thermal regime established by the energy in the Earth's interior. This component of the signal is estimated as a linear increase in temperature with depth from the lower section of a borehole temperature profile, which is assumed to be unperturbed by recent changes in climate at the surface. The validity of this assumption is dependent on both the subsurface thermophysical properties and the character of the downwelling climatic signal. Such uncertainties can significantly impact the determination of the quasi-steady state thermal regime, and consequently the magnitude of the temperature anomaly interpreted as a climatic signal. The quantitative effects and uncertainties that arise from the analysis of temperature-depth profiles of different depths are presented. Results demonstrate that widely different GST histories can be derived from a single temperature profile truncated at different depths. Borehole temperature measurements approaching 500-600 m depths are shown to provide the most robust GST reconstructions spanning 500 to 1000 yr BP. It is further shown that the bias introduced by a temperature profile of depths shallower than 500-600 m remains even if the time span of the reconstruction target is shortened

Increasingly transformed MCF-10A cells have a progressively tumor-like phenotype in three-dimensional basement membrane culture

<p>Abstract</p> <p>Background</p> <p>MCF-10A cells are near diploid and normal human mammary epithelial cells. In three-dimensional reconstituted basement membrane culture, they undergo a well-defined program of proliferation, differentiation, and growth arrest, forming acinar structures that recapitulate many aspects of mammary architecture <it>in vivo</it>. The pre-malignant MCF-10AT cells and malignant MCF-10CA1a lines were sequentially derived from the MCF-10A parental cell line first by expression of a constitutively active T24 H-Ras generating the MCF-10AT cell line. This was followed by repeated selection for increasingly aggressive tumor formation from cells recovered from xenograft tumors in immuno-compromised mice, generating the MCF-10CA1a cell line. When inoculated subcutaneously into the flanks of immuno-compromised mice, MCF-10AT cells occasionally form tumors, whereas MCF-10CA1a cells invariably form tumors with a shorter latency than MCF-10AT derived tumors.</p> <p>Results</p> <p>MCF-10AT cells grown in three-dimensional basement membrane culture form complex multi-acinar structures that produce a basement membrane but undergo delayed cell cycle arrest and have incomplete luminal development. MCF-10CA1a cells grown in three-dimensional basement membrane culture form large, hyper-proliferative masses, that retain few characteristics of MCF10A acini and more closely resemble tumors.</p> <p>Conclusion</p> <p>Here we report on the growth and differentiation properties of these three matched cell lines in three-dimensional basement membrane culture. Features of tissue morphogenesis were assessed, including proliferation, basement membrane formation, polarization of alpha-6 beta-4 integrin to the basement membrane, formation of cell:cell junctions, and apoptosis for luminal clearance. The matched series of normal MCF-10A, pre-malignant MCF-10AT, and malignant MCF-10CA1a cells offers a unique opportunity to study the mechanisms of malignant progression both in a three-dimensional microenvironment and in the same cell background.</p

Molecular evolution of urea amidolyase and urea carboxylase in fungi

Background: Urea amidolyase breaks down urea into ammonia and carbon dioxide in a two-step process, while another enzyme, urease, does this in a one step-process. Urea amidolyase has been found only in some fungal species among eukaryotes. It contains two major domains: the amidase and urea carboxylase domains. A shorter form of urea amidolyase is known as urea carboxylase and has no amidase domain. Eukaryotic urea carboxylase has been found only in several fungal species and green algae. In order to elucidate the evolutionary origin of urea amidolyase and urea carboxylase, we studied the distribution of urea amidolyase, urea carboxylase, as well as other proteins including urease, across kingdoms. Results: Among the 64 fungal species we examined, only those in two Ascomycota classes (Sordariomycetes and Saccharomycetes) had the urea amidolyase sequences. Urea carboxylase was found in many but not all of the species in the phylum Basidiomycota and in the subphylum Pezizomycotina (phylum Ascomycota). It was completely absent from the class Saccharomycetes (phylum Ascomycota; subphylum Saccharomycotina). Four Sordariomycetes species we examined had both the urea carboxylase and the urea amidolyase sequences. Phylogenetic analysis showed that these two enzymes appeared to have gone through independent evolution since their bacterial origin. The amidase domain and the urea carboxylase domain sequences from fungal urea amidolyases clustered strongly together with the amidase and urea carboxylase sequences, respectively, from a small number of beta- and gammaproteobacteria. On the other hand, fungal urea carboxylase proteins clustered together with another copy of urea carboxylases distributed broadly among bacteria. The urease proteins were found in all the fungal species examined except for those of the subphylum Saccharomycotina. Conclusions: We conclude that the urea amidolyase genes currently found only in fungi are the results of a horizontal gene transfer event from beta-, gamma-, or related species of proteobacteria. The event took place before the divergence of the subphyla Pezizomycotina and Saccharomycotina but after the divergence of the subphylum Taphrinomycotina. Urea carboxylase genes currently found in fungi and other limited organisms were also likely derived from another ancestral gene in bacteria. Our study presented another important example showing plastic and opportunistic genome evolution in bacteria and fungi and their evolutionary interplay

1865-09-24 N.E. Nickerson requests Adjutant General Hodsdon inform the commanding officer at Fort Independence that the regiment has been mustered out

https://digitalmaine.com/cw_me_1st_heavy_corr/1439/thumbnail.jp

Evaluation of Fat-free Mass Characteristics at Different Adiposity Levels: Impact of Weight Status Stratification Method

The method used to stratify weight status when evaluating fat-free mass (FFM) characteristics has primarily been based upon body mass index (BMI) and not body fat percent (BF%). As a result, it is unknown whether deviations in FFM characteristics are similar when stratifying weight status based upon BMI or BF%. Purpose: The purpose of this study was to evaluate FFM characteristics at different adiposity levels when stratifying weight status via BMI and BF%. Methods: 150 adults (50% males) participated in this study. 3-compartment (3C) model body composition was based upon body density (Db) via air displacement plethysmography (ADP) and total body water via bioimpedance analysis. FFM density (DFFM), residual (RFFM), and hydration (HFFM)were evaluated in all subjects. The Db obtained from a stand-alone assessment of ADP (2C-Db) and 3C model (3C-Db)were also compared between groups. Subjects were stratified based upon weight status (BMI and BF%). First, subjects were divided into a normal weight (NW-BMI), overweight (OW-BMI) or obese (OB-BMI) group, which were based upon BMI values of 18.5-24.99 kg/m2 (n=50), 25.0-29.99 kg/m2 (n=50), and ³ 30.0 kg/m2 (n=50), respectively. Next, subjects were divided into groups based upon BF% where normal weight BF% values (NW-BF%) for males and females were \u3c 25 and 35% (n=81), respectively, and obese BF% values (OB-BF%) were ³25 and 35% (n=69), respectively. Results: The DFFM ranged from 1.096 – 1.097 g/cm3 for all groups in both weight status stratification methods (all p \u3e 0.05). HFFM and RFFM were similar for all comparisons and ranged from 73.99 – 74.33% and 25.67 – 26.01%, respectively, for BMI groups and 74.02 – 74.26% and 25.74 – 25.98%, respectively, for BF% groups (all p \u3e 0.05). In contrast, the 3C-Db (1.050, 1.036, and 1.013 g/cm3) and 2C-Db (1.051, 1.036, and 1.014 g/cm3) were statistically significant for all comparisons between NW-BMI, OW-BMI, and OB-BMI respectively (all p \u3c 0.05). Furthermore, 3C-Db (1.051 and 1.011 g/cm3) and 2C-Db (1.052 and 1.012 g/cm3) were significantly different when comparing NW-BF% and OB-BF% (both p \u3c 0.05). CONCLUSIONS: Previous research has reported the FFM characteristics when stratifying weight status via BMI classification. As a result, it was unknown whether deviations in FFM characteristics existed when stratifying by BF%. Uniquely, the current study findings revealed that FFM characteristics are similar between groups regardless of the weight status stratification method (BMI or BF%)