Gap junction structures: Analysis of the x-ray diffraction data

Models for the spatial distribution of protein, lipid and water in gap junction structures have been constructed from the results of the analysis of X-ray diffraction data described here and the electron microscope and chemical data presented in the preceding paper (Caspar, D. L. D., D. A. Goodenough, L. Makowski, and W.C. Phillips. 1977. 74:605-628). The continuous intensity distribution on the meridian of the X-ray diffraction pattern was measured, and corrected for the effects of the partially ordered stacking and partial orientation of the junctions in the X-ray specimens. The electron density distribution in the direction perpendicular to the plane of the junction was calculated from the meridional intensity data. Determination of the interference function for the stacking of the junctions improved the accuracy of the electron density profile. The pair-correlation function, which provides information about the packing of junctions in the specimen, was calculated from the interference function. The intensities of the hexagonal lattice reflections on the equator of the X-ray pattern were used in coordination with the electron microscope data to calculate to the two-dimensional electron density projection onto the plane of the membrane. Differences in the structure of the connexons as seen in the meridional profile and equatorial projections were shown to be correlated to changes in lattice constant. The parts of the junction structure which are variable have been distinguished from the invariant parts by comparison of the X-ray data from different specimens. The combination of these results with electron microscope and chemical data provides low resolution three- dimensional representations of the structures of gap junctions.     

Acute inflammation and infection maintain circulating phospholipid levels and enhance lipopolysaccharide binding to plasma lipoproteins

Circulating lipoproteins are thought to play an important role in the detoxification of lipopolysaccharide (LPS) by binding the bioactive lipid A portion of LPS to the lipoprotein surface. It has been assumed that hypocholesterolemia contributes to inflammation during critical illness by impairing LPS neutralization. We tested whether critical illness impaired LPS binding to lipoproteins and found, to the contrary, that LPS binding was enhanced and that LPS binding to the lipoprotein classes correlated with their phospholipid content. Whereas low serum cholesterol was almost entirely due to the loss of esterified cholesterol (a lipoprotein core component), phospholipids (the major lipoprotein surface lipid) were maintained at near normal levels and were increased in a hypertriglyceridemic subset of septic patients. The levels of phospholipids found in the LDL and VLDL fractions varied inversely with those in the HDL fraction, and LPS bound predominantly to lipoproteins in the LDL and VLDL fractions when HDL levels were low. Lipoproteins isolated from the serum of septic patients neutralized the bioactivity of the LPS that had bound to them. Our results show that the host response to acute inflammation and infection tends to maintain lipoprotein phospholipid levels and that, despite hypocholesterolemia and reduced HDL levels, circulating lipoproteins maintain their ability to bind and neutralize an important bacterial agonist, LPS.     

Role of thromboxane in producing portal hypertension following trauma-hemorrhage

Thromboxane A2 (TXA2) and endothelin-1 (ET-1) have been proposed as the important vasoconstrictors that increase portal venous resistance in paracrine or autocrine fashion. We hypothesized that the hepatic damage following trauma-hemorrhage (T-H) is induced by the impaired hepatic circulation due to the increased production of vasoconstrictors such as ET-1 and TXA2 by the liver. To test this, male Sprague-Dawley rats (n = 6/group) were subjected to trauma (i.e., midline laparotomy) and hemorrhage (35-40 mmHg for 90 min followed by fluid resuscitation) or sham operation. At 2 or 5 h after the end of resuscitation, the liver was isolated and perfused and portal inflow pressure, bile flow, and release of ET-1 and thromboxane B2 (TXB2; a stable metabolite of TXA2) into the perfusate were measured. The level of portal pressure was higher at 5 h following T-H compared with 2 h after T-H and sham. The portal pressure was inversely correlated to the amount of bile production. Furthermore, the bile flow was significantly correlated to the hepatic damage as evidenced by release of lactate dehydrogenase into the perfusate. The level of ET-1 at 5 h following T-H in the perfusate after 30 min of recirculation did not show any difference from sham. However, the levels of TXB2 in the T-H group were significantly higher than those in sham at that interval. These results indicate that the increased release of TXA2 but not ET-1 following T-H might be responsible for producing the increased portal resistance, decreased bile production, and hepatic damage.     

Natal location influences movement and survival of a spatially structured population of snail kites

Despite the accepted importance of the need to better understand how natal location affects movement decisions and survival of animals, robust estimates of movement and survival in relation to the natal location are lacking. Our study focuses on movement and survival related to the natal location of snail kites in Florida and shows that kites, in addition to exhibiting a high level of site tenacity to breeding regions, also exhibit particular attraction to their natal region. More specifically, we found that estimates of movement from post-dispersal regions were greater toward natal regions than toward non-natal regions (differences were significant for three of four regions). We also found that estimates of natal philopatry were greater than estimates of philopatry to non-natal regions (differences were statistically significant for two of four regions). A previous study indicated an effect of natal region on juvenile survival; in this study, we show an effect of natal region on adult survival. Estimates of adult survival varied among kites that were hatched in different regions. Adults experienced mortality rates characteristic of the region occupied at the time when survival was measured, but because there is a greater probability that kites will return to their natal region than to any other regions, their survival was ultimately influenced by their natal region. In most years, kites hatched in southern regions had greater survival probabilities than did kites hatched in northern regions. However, during a multiregional drought, one of the northern regions served as a refuge from drought, and during this perturbation, survival was greater for birds hatched in the north. Our study shows that natal location may be important in influencing the ecological dynamics of kites but also highlights the importance of considering temporal variation in habitat conditions of spatially structured systems when attempting to evaluate the conservation value of habitats. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Vegetation change from chronic stress events: Detection of the effects of tide gate removal and long‐term drought on a tidal marsh

Question: Chronic stress events are defined as disturbance events that exceed the lifespan of the dominant plant species, fluctuate in intensity and lack abruptness or physical destruction of biomass. Can the effects of chronic stress events be measured on vegetation communities? Did two chronic stress events, the removal of a tide gate and a four year drought, cause a temporary or permanent shift in the vegetation communities of a tidal marsh? Location: Tidal marsh in southeastern United States. Methods: Change in species composition and dominance and community change on a landscape level salinity gradient were measured between time periods ranging from four months to seven years to construct a statistical baseline reference community at freshwater, oligohaline, and mesohaline sections of a tidal marsh. Statistical shifts in the plant community were defined as changes in the plant community that fell outside of the defined baseline reference community. Results: Plant community changes outside of the reference community occurred in 13 out of 378 community comparisons. Removal of the tide gate had a greater effect on interstitial salinity levels than the drought and was most intense in the oligohaline marsh, where between 20 to 45% of the freshwa‐ter/oligohaline community types permanently converted to oligohaline community types. However, community shifts in the freshwater and oligohaline marsh induced by the drought were temporary, lasting from 1 to 3+ years. Neither chronic stress event permanently altered the mesohaline plant communities. Conclusion: The effects of chronic stress events could be detected; an extended historical record of vegetation change (18 years) was necessary to identify community shifts outside of a reference condition of the community and to determine if those shifts were permanent or temporary.     

NK cells can trigger allograft vasculopathy: the role of hybrid resistance in solid organ allografts

Progressive arterial stenosis (cardiac allograft vasculopathy (CAV)) is a leading cause of long-term failure of organ transplants. CAV remains intractable, in part because its mechanisms are insufficiently understood. A central proposition is that MHC-driven alloimmune processes play a necessary role in CAV, as shown by the absolute requirement for histoincompatibility between donor and recipient for its production. Two immunological pathways have been implicated involving reactivity to donor MHC Ags by either T or B cells. In this study, we use a novel system of semiallogeneic cardiac transplants between parental donors and F1 hybrid recipients to provide evidence that NK cells, members of the innate immune system, also contribute to the generation of CAV in mice. This finding marks the first demonstration that the hybrid resistance phenomenon occurs in solid organ allografts. Extension of these experiments to recipients deficient in T cells demonstrates that this third pathway of CAV, the NK cell-triggered pathway, involves the recruitment of T cells not responsive to donor alloantigens. Finally, transplants performed with donors or recipients deficient in IFN-gamma revealed that recipient-derived IFN-gamma is necessary for CAV formation in parental to F1 transplants, suggesting a possible effector mechanism by which NK cells can promote CAV. Together, these results define a previously unknown pathway toward CAV and assign a novel role to NK cells in organ allograft rejection.     

The role of nitric oxide in cancer

Nitric oxide (NO) is a pleiotropic regulator, critical to numerous biological processes, including va-sodilatation, neurotransmission and macrophage-mediated immunity. The family of nitric oxide synthases (NOS) comprises inducible NOS (iNOS), endothelial NOS (eNOS), and neuronal NOS (nNOS). Interestingly, various studies have shown that all three isoforms can be involved in promoting or inhibiting the etiology of cancer. NOS activity has been detected in tumour cells of various histogenetic origins and has been associated with tumour grade, proliferation rate and expression of important signaling components associated with cancer development such as the oestrogen receptor. It appears that high levels of NOS expression (for example, generated by activated macrophages) may be cytostatic or cytotoxic for tumor cells, whereas low level activity can have the opposite effect and promote tumour growth. Paradoxically therefore, NO (and related reactive nitrogen species) may have both genotoxic and angiogenic pro