Emergent role of the fractalkine axis in dissemination of peritoneal metastasis from epithelial ovarian carcinoma.

Epithelial ovarian carcinoma is the most common cause of death from gynecologic cancers largely due to advanced, relapsed and chemotherapy-resistant peritoneal metastasis, which is refractory to the currently used treatment approaches. Mechanisms supporting advanced and relapsed peritoneal metastasis are largely unknown, precluding development of more effective targeted therapies. In this study, we investigated the function of a potentially targetable fractalkine axis in the formation and the development of advanced and relapsed peritoneal metastasis and its impact on patients' outcomes. Our mouse model studies support a role for the fractalkine receptor (CX3CR1) in the initiation of peritoneal adhesion important for recolonization of relapsed peritoneal metastasis. We show that downregulation of CX3CR1 results in reduction of metastatic burden at several peritoneal sites commonly colonized by advanced and relapsed metastatic ovarian carcinoma. We show that the chemokine fractalkine (CX3CL1), an activating ligand of CX3CR1, regulates organ-specific peritoneal colonization. High expression of CX3CR1 correlates with significantly shorter survival, specifically in post-menopausal patients with advanced and terminal stages of the disease. Taken together, our studies support a key regulatory role for the fractalkine axis in advanced and relapsed peritoneal metastasis in epithelial ovarian carcinoma.

Focal adhesion kinase and beta1 integrin regulation of Na+, K+, 2Cl- cotransporter in osmosensing ion transporting cells of killifish, Fundulus heteroclitus.

Focal adhesion kinase (FAK), also known as PYK2, is a tyrosine kinase that functions in integrin-mediated signaling in mechanosensitive cells but its role in osmosensing cells is unknown. Antibodies directed against phosphorylated FAK, whose epitopes are conserved among vertebrates, were used to follow phosphorylation patterns in an osmosensing ion secreting epithelium, the killifish (Fundulus heteroclitus) opercular membrane. At the electron microscopic level, a unique combination of integrin beta1, the phosphorylated form of FAK at tyrosine 407 (pY407) and Na(+), K(+), 2Cl(-) cotransporter (NKCC1) were all colocalized only on the basolateral membrane in chloride cells. The three proteins were also coimmunoprecipitated with each other in isotonic conditions, suggesting an osmosensing complex involving the three proteins. Only FAK pY407 was sensitive to hypotonic shock and became dephosphorylated with hypotonic shock, while FAK pY576 in the apical membrane and pY861 in cell-cell adhesions were insensitive to hypotonicity. NKCC1 contributes to NaCl secretion in seawater and previous reports showed that hypotonic shock (-60 mOsm/kg) rapidly inhibits Cl(-) secretion. These results indicate that chloride cells respond to hypotonic shock using integrin beta1 as an osmosensor that is connected to dephosphorylation of FAK pY407 which leads to NKCC1 deactivation in the basolateral membrane and the inhibition of NaCl secretion by these epithelial cells.

Atmospheric volatile organic compound measurements during the 1996 Paso del Norte Ozone Study.

Ambient air VOC samples were collected at surface air quality monitoring sites, near sources of interest, and aloft on the US (El Paso) and Mexican (Ciudad Juárez) side of the border during a six-week period of the 1996 Paso del Norte Ozone Study. Samples were collected at five sites, three on the US side and two on the Mexican side, during nine intensive operation days when high ozone levels were forecast for the area. Six other sites were sampled to characterize up-wind, down-wind and other emission sources. Samples for determining source profiles were collected for rush hour traffic, propane-powered bus exhaust, automobile paint shop emissions, propane fuels, and industrial manufacturing in Cd. Juárez and a refinery in El Paso. Most samples were collected in electro-polished stainless steel canisters for determination of C2 to C(10+) hydrocarbons by GC-FID. Carbonyl samples were collected on DNPH impregnated cartridges at three surface sites during aircraft flights and analyzed by HPLC. This paper presents the spatial and temporal characteristics of VOC species concentrations and compositions to examine the differences and similarities of the various locations and time periods. Overall surface, total non-methane hydrocarbon values ranged from 0.1 to 3.4 ppmC with the highest concentrations being recorded in the morning and evening at five vehicle-dominated sites, three in Cd. Juárez and two in El Paso. Toluene in El Paso samples and propane, which is used as a cooking and transportation fuel in Cd. Juárez, were the most abundant hydrocarbons. The most abundant carbonyls were acetaldehyde, acetone and formaldehyde.

The 1996 Paso del Norte Ozone Study: analysis of meteorological and air quality data that influence local ozone concentrations.

The 1996 Paso del Norte Ozone Study and subsequent data analyses were implemented to develop an understanding of the chemical and physical processes which lead to high concentrations of ozone in the Paso del Norte study area which includes El Paso County, Texas, Sunland Park, New Mexico, and Ciudad Juárez, Mexico. Both the data and data analysis results are being used to support photochemical grid modeling. El Paso County and Sunland Park fail to meet the National Ambient Air Quality Standard (NAAQS) for ozone, and neighboring Ciudad Juárez fails to meet the Mexican ambient standard for ozone. This paper summarizes the measurement campaigns of the 1996 Paso del Norte Ozone Study and the findings and conclusions that arose from subsequent data analyses. Data analyses showed that high ozone concentrations resulted from a combination of conditions, including high surface temperatures, strong sunlight with few clouds, light surface winds and high concentrations of ozone precursors at ground level in the morning, and slow convective boundary layer (CBL) growth. Synoptic-scale meteorological conditions observed during high ozone episodes included an aloft high-pressure system and aloft warming. Aloft carryover of ozone and ozone precursors did not significantly contribute to high concentrations of ozone at the surface.

Differentiation of Y79 cells induced by prolonged exposure to insulin.

Y79 human retinoblastoma cells are known to contain receptors for both insulin and insulin-like growth factors (IGFs), to produce these cytokines and release them in the culture medium. Previously we have demonstrated that IGFs and insulin stimulate Y79 cell proliferation through the involvement of type I IGF receptor and Insulin Receptor Substrate 1 (IRS-1). This paper studies the effect of prolonged exposure to insulin on Y79 cells. Cells grown for 10 days in the presence of insulin were reseeded and incubated once more with insulin. In the reseeded cells proliferation lowered and morphological changes appeared. After 10 days of reseeding, cells stopped proliferating and showed long ramifying neurite processes and varicosities consistent with neuronal differentiation. Morphological differentiation was accompanied by a marked increase in the content of total protein and in that of tubulin, the major protein constituent of microtubules, a marked increase in the content of specialized protein markers of dopaminergic and cholinergic differentiation (dopamine beta-hydroxylase and choline acetyltransferase activities, respectively); a contemporaneous decrease in the content of glial fibrillary acidic protein (GFAP), a specific marker of glial cells, was also observed. Our results demonstrate that prolonged exposure to insulin induces Y79 cells to differentiate into a neuronal-like phenotype. At this moment it is not possible to establish the mechanism by which insulin induces this differentiative effect.

Insulin and IGFs induce apoptosis in chick embryo retinas deprived of L-glutamine.

In chick embryo retinas, cultured in serum-free medium lacking L-glutamine, IGF-I, IGF-II and insulin induced apoptotic DNA fragmentation and cell death, IGF-I being the most efficacious compound. The apoptotic effect, which was particularly evident in retinas removed from 7-day-old chick embryos, declined with the age of the embryos and disappeared after day 11. Apoptosis appeared after a time lag of 8 h and then increased with time up to 16 h. Cycloheximide, an inhibitor of protein synthesis, was capable of entirely abolishing apoptotic cell death. The effect induced by IGFs or insulin was suppressed by the addition of glutamine. Cytokine-mediated apoptosis was also observed after withdrawal of phosphate. We suggest that IGFs or insulin may produce, in retinas cultured in medium lacking L-glutamine or phosphate, a conflict of signals that could be lethal for retinal cells.

Changes in protein composition during chick embryo retina development.

Two dimensional gel electrophoresis was employed to analyse the protein composition of chick embryo retina during "in ovo" development in order to individuate some components which are typical of different stages of growth. The study of protein composition of 7-day-old chick embryo retinas revealed the presence of about eighty different soluble components. In four of these proteins the staining intensity decreased during retina development. One of these components, of 35 KD with an isoelectric point of about 5.15, was partially purified by means of preparative isoelectric focusing. Other components, which were absent at day 7 appeared during the second week of development and were clearly visualised at day 16. These results were also compared with those obtained by means of 2DGE performed on liver and brain of chick embryo at day 7 and 16 of development.

Cumulative trauma disorders among apparel manufacturing employees in the southeastern United States.

Factors associated with cumulative trauma disorders (CTDs) of the upper extremities were studied retrospectively, using data from three apparel manufacturing plants in the southeastern United States. Reported CTDs among the employees at risk during fiscal years 1991 and 1992 were analyzed focusing on individual and plant variables. Results showed that CTD rates increased from 1991 to 1992 and fluctuated in a cyclical pattern each year, peaking during January, March, and August. Employee age and duration of employment were associated with CTD rates. Employees aged 45 to 49 had higher CTD rates than those in any other age groups. Employees with 1 to 3 years' experience had higher CTD rates than employees with more years of service.