Cytokeratins of Tumorigenic and Highly Malignant Respiratory Tract Epithelial Cells

In malignant airway epithelial cells, structural abnormalities were evident from the cytokeratin organization. To determine whether the cytokeratins themselves were responsible, an in vitro model for bronchogenic carcinoma, consisting of three highly malignant lines and three less tumorigenic lines, was studied. Cytokeratins were evaluated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). When typical constraints on tumors were relieved by in vitro culture, lines showed profiles resembling normal, primary cells. The CK5/CK14 combination, characteristic of basal epithelial layers, was represented by CK6A/CK14. CK17 was invariably present, while CK5, CK7, CK8, CK19, and CK42 content varied. CK19 appeared to substitute for the rarely observed CK18. While lacking the common CK8/CK18 combination of hyperproliferative cells, an invasive, metastasizing line had CK6A/CK7 or CK8 with CK19 suggesting derivation similar to adenocarcinomas. Bands of CK19 and actin migrated to higher pI in tumorigenic and malignant lines than in normal cells. Ubiquitinated acidic cytokeratins with a low isoelectric point (pI) and high molecular weight (MW) showed no consistent differences in lines that differed in growth potential. Type II made up 49–52% of total cytokeratins in nonmalignant lines, whereas highly malignant lines showed lower levels. Posttranslational modifications were identified but could not explain the shortfall of basic cytokeratins

Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor

Protein kinase Cs (PKCs) are activated by lipids in the plasma membrane and bind to a scaffold assembled on the epidermal growth factor (EGF) receptor (EGFR). Understanding how this complex is routed is important, because this determines whether EGFR is degraded, terminating signaling. Here, cells were preincubated in EGF-tagged gold nanoparticles, then allowed to internalize them in the presence or absence of a phorbol ester PKC activator. PKC colocalized with EGF-tagged nanoparticles within 5 min and migrated with EGFR-bearing vesicles into the cell. Two conformations of PKC-epsilon were distinguished by different primary antibodies. One, thought to be enzymatically active, was on endosomes and displayed a binding site for antibody RR (R&D). The other, recognized by Genetex green (GG), was soluble, on actin-rich structures, and loosely bound to vesicles. During a 15-min chase, EGF-tagged nanoparticles entered large, perinuclear structures. In phorbol ester-treated cells, vesicles bearing EGF-tagged nanoparticles tended to enter this endocytic recycling compartment (ERC) without the GG form. The correlation coefficient between the GG (inactive) and RR conformations on vesicles was also lower. Thus, active PKC has a Charon-like function, ferrying vesicles to the ERC, and inactivation counteracts this function. The advantage conferred on cells by aggregating vesicles in the ERC is unclear

Unraveling the Determinants of Protrusion Formation

A computerized morphometric classification technique based on latent factors reveals major protrusion classes: factors 4, 5, and 7. Previous work showed that factor 4 represented filopodia, 5 the distribution of lamellar cytoplasm, and 7 a blunt protrusion. We explore the relationship of focal contact (FC) characteristics and their integrated actin cables to factors values. The results show that FC maturation/cytoskeletal integration affects factor 5, because FC elongation/integration was correlated with its values. On the contrary, 7 values decreased with maturation, so cable or FC size or their integration must be restricted to form these protrusions. Where integration did occur, the cables showed distinctive size and orientation, as indicated by correlation of 7 values with FC shape. Results obtained with myosin inhibitors support the interpretation that a central, isometric, contractile network puts constraints on both factor 5 and 7 protrusions. We conclude that cells establish functional domains by rearranging the cytoskeleton

Pattern Analysis of Microtubule-Polymerizing and -Depolymerizing Agent Combinations as Cancer Chemotherapies

Subcellular distribution of mass can be analyzed by a technique that involves culturing cells on interferometers and digitizing their interference contours. Contour sampling resulted in 102 variables per cell, which were predictors of oncogenic transformation. Cell phenotypes can be deconstructed by use of latent factors, which represent the covariance of the real variables. The reversal of the cancertype phenotype by a combination of microtubule- stabilizing and -depolymerizing agents was described previously. The implications of these results have been explored by clinicians who treated patients with the combination of docetaxel and vinorelbine (Navelbine®). The current study was performed to determine the effects of different combinations on phenotype and in phases of the cell cycle other than mitosis. Combinations of paclitaxel with either colchicine, podophyllotoxin, nocodazole, or vinblastine caused phenotype reversal. Paclitaxel analogue, 7-deoxytaxol, by itself caused reversal. Factors #4, (filopodia), #5 (displacement and/or deep invaginations in the periphery), #8, and #12 took on values typical of normal cells, whereas the values of #7 (p21-activated kinase), and #13 (rounding up) shifted toward the cancer-type. All combinations altered microtubule arrangement at the cell edge. Delivery schedules and drug ratios used in clinical studies were subjected to analysis. Clinical response rates were better when the combination was not interspersed with a single agent (P=0.004). The results support the idea that efficacy depends upon simultaneous exposure to both agents, and suggest a novel mechanism for combination therapies. These therapies appear to restore in transformed cells some of the features of a contact-inhibited cell, and to impede progress through the cell cycle even when provided at nanomolar concentrations

Expression of glycolytic enzymes in ovarian cancers and evaluation of the glycolytic pathway as a strategy for ovarian cancer treatment

Table S2. Spearman correlation of the expression of four glycolytic enzymes in a cohort of 380 ovarian cancers. Spearman rho correlation values (top value) along with the respective adjusted P value (bottom value) of statistically significant correlations thresholded at FDR P < 0.01 are summarised. (DOCX 21 kb

Insomnia symptoms and repressive coping in a sample of older Black and White women

BACKGROUND: This study examined whether ethnic differences in insomnia symptoms are mediated by differences in repressive coping styles. METHODS: A total of 1274 women (average age = 59.36 ± 6.53 years) participated in the study; 28% were White and 72% were Black. Older women in Brooklyn, NY were recruited using a stratified, cluster-sampling technique. Trained staff conducted face-to-face interviews lasting 1.5 hours acquiring sociodemographic data, health characteristics, and risk factors. A sleep questionnaire was administered and individual repressive coping styles were assessed. Fisher's exact test and Spearman and Pearson analyses were used to analyze the data. RESULTS: The rate of insomnia symptoms was greater among White women [74% vs. 46%; χ(2 )= 87.67, p < 0.0001]. Black women scored higher on the repressive coping scale than did White women [Black = 37.52 ± 6.99, White = 29.78 ± 7.38, F(1,1272 )= 304.75, p < 0.0001]. We observed stronger correlations between repressive coping and insomnia symptoms for Black [r(s )= -0.43, p < 0.0001] than for White women [r(s )= -0.18, p < 0.0001]. Controlling for variation in repressive coping, the magnitude of the correlation between ethnicity and insomnia symptoms was substantially reduced. Multivariate adjustment for differences in sociodemographics, health risk factors, physical health, and health beliefs and attitudes had little effect on the relationships. CONCLUSION: Relationships between ethnicity and insomnia symptoms are jointly dependent on the degree of repressive coping, suggesting that Black women may be reporting fewer insomnia symptoms because of a greater ability to route negative emotions from consciousness. It may be that Blacks cope with sleep problems within a positive self-regulatory framework, which allows them to deal more effectively with sleep-interfering psychological processes to stressful life events and to curtail dysfunctional sleep-interpreting processes

Ultraluminous Infrared Galaxies

Ever since their discovery in the 1970's, UltraLuminous InfraRed Galaxies (ULIRGs; classically Lir>10^12Lsun) have fascinated astronomers with their immense luminosities, and frustrated them due to their singularly opaque nature, almost in equal measure. Over the last decade, however, comprehensive observations from the X-ray through to the radio have produced a consensus picture of local ULIRGs, showing that they are mergers between gas rich galaxies, where the interaction triggers some combination of dust-enshrouded starburst and AGN activity, with the starburst usually dominating. Very recent results have thrown ULIRGs even further to the fore. Originally they were thought of as little more than a local oddity, but the latest IR surveys have shown that ULIRGs are vastly more numerous at high redshift, and tantalizing suggestions of physical differences between high and low redshift ULIRGs hint at differences in their formation modes and local environment. In this review we look at recent progress on understanding the physics and evolution of local ULIRGs, the contribution of high redshift ULIRGs to the cosmic infrared background and the global history of star formation, and the role of ULIRGs as diagnostics of the formation of massive galaxies and large-scale structures.Comment: Review article, published in "Astrophysics Update 2 - topical and timely reviews on astronomy and astrophysics". Ed. John W. Mason. Springer/Praxis books. ISBN: 3-540-30312-X. 53 pages, 5 figures. Higher quality figures available on reques