In situ assays demonstrate that interferon-gamma suppresses infection-stimulated hepatic fibrin deposition by promoting fibrinolysis.

BACKGROUND: Inflammatory cytokines potently impact hemostatic pathways during infection, but the tissue-specific regulation of coagulation and fibrinolysis complicates studies of the underlying mechanisms. METHODS AND RESULTS: Here, we describe assays that quantitatively measuring prothrombinase (PTase), protein C-ase (PCase) and plasminogen activator (PA) activities in situ, thereby facilitating studies of tissue-specific hemostasis. Using these assays, we investigate the mechanisms regulating hepatic fibrin deposition during murine toxoplasmosis and the means by which interferon-gamma (IFN-gamma) suppresses infection-stimulated fibrin deposition. We demonstrate that Toxoplasma infection upregulates hepatic PTase, PCase, and PA activity. Wild type and gene-targeted IFN-gamma-deficient mice exhibit similar levels of infection-stimulated PTase activity. By contrast, IFN-gamma-deficiency is associated with increased PCase activity and reduced PA activity during infection. Parallel analyses of hepatic gene expression reveal that IFN-gamma-deficiency is associated with increased expression of thrombomodulin (TM), a key component of the PCase, increased expression of thrombin-activatable fibrinolysis inhibitor (TAFI), a PC substrate, and reduced expression of urokinase PA (u-PA). CONCLUSIONS: These findings suggest that IFN-gamma suppresses infection-stimulated hepatic fibrin deposition by suppressing TM-mediated activation of TAFI, thereby destabilizing fibrin deposits, and concomitantly increasing hepatic u-PA activity, thereby promoting fibrinolysis. We anticipate that further application of these in situ assays will improve our understanding of tissue-specific hemostasis, its regulation by cytokines, and its dysregulation during coagulopathy.

Interferon-gamma is crucial for surviving a Brucella abortus infection in both resistant C57BL/6 and susceptible BALB/c mice.

Brucella abortus is an intracellular bacterial pathogen that causes chronic infections in humans and a number of agriculturally important species of animals. It has been shown that BALB/c mice are more susceptible to infections with virulent strains of Brucella abortus than C57BL/6 or C57BL/10 strains. In experiments described here, gene knock-out mice were utilized to elucidate some of the salient components of resistance. Resistant C57BL/6 mice with gene deletions or disruptions in the interferon-gamma (IFN-gamma), perforin or beta(2)-microglobulin genes had decreased abilities to control intracellular infections with B. abortus strain 2308 during the first week after infection. However, only the IFN-gamma knock-out mice had a sustained inability to control infections and this resulted in death of the mice at approximately 6 weeks post-infection. These mice had a continual increase in the number of bacterial colony-forming units (CFU) in their spleens until death. When BALB/c mice with the disrupted IFN-gamma gene were infected they had more splenic CFU at one week post-infection than control mice but the increase was not statistically significant and by 3 weeks they did not have more CFU than control mice. Moreover, the number of splenic bacteria did not increase in the BALB/c IFN-gamma knock-out mice between 6 and 10.5 weeks, although they died at 10.5 weeks, the time by which normal BALB/c mice were clearing the infection. Death in both strains of IFN-gamma gene disrupted mice coincided with symptoms of cachexia and macrophages comprised > or= 75% of the splenic leucocytes.

dnaC mutations suppress defects in DNA replication- and recombination-associated functions in priB and priC double mutants in Escherichia coli K-12.

PriA, PriB and PriC were originally discovered as proteins essential for the PhiX174 in vitro DNA replication system. Recent studies have shown that PriA mutants are poorly viable, have high basal levels of SOS expression (SOSH), are recombination deficient (Rec-), sensitive to UV irradiation (UVS) and sensitive to rich media. These data suggest that priA's role may be more complex than previously thought and may involve both DNA replication and homologous recombination. Based on the PhiX174 system, mutations in priB and priC should cause phenotypes like those seen in priA2:kan mutants. To test this, mutations in priB and priC were constructed. We found that, contrary to the PhiX174 model, del(priB)302 and priC303:kan mutants have almost wild-type phenotypes. Most unexpectedly, we then found that the priBC double mutant had very poor viability and/or a slow growth rate (even less than a priA2:kan mutant). This suggests that priB and priC have a redundant and important role in Escherichia coli. The priA2:kan suppressor, dnaC809, partially suppressed the poor viability/slow growth phenotype of the priBC double mutant. The resulting triple mutant (priBC dnaC809 ) had small colony size, recombination deficiency and levels of SOS expression similar to a priA2:kan mutant. The priBC dnaC809 mutant, however, was moderately UVR and had good viability, unlike a priA2:kan mutant. Additional mutations in the triple mutant were selected to suppress the slow growth phenotype. One suppressor restored all phenotypes tested to nearly wild-type levels. This mutation was identified as dnaC820 (K178N) [mapping just downstream of dnaC809 (E176G)]. Experiments suggest that dnaC820 makes dnaC809 suppression of priA and or priBC mutants priB and or priC independent. A model is proposed for the roles of these proteins in terms of restarting collapsed replication forks from recombinational intermediates.

Analytic solution for the peak reflectivity of an asymmetric mirror.

Asymmetric Bragg reflectors have been shown to optimize mirror performance in strained-layer material systems. Although the theory behind the reflectivity of symmetric mirrors has been well studied, little is known about asymmetric mirror designs. We present a closed-form solution for the peak reflectivity of an asymmetric mirror that results from applying a tanh substitution. This elegant technique has been shown to yield a markedly simplified calculation of the peak reflectivity of a symmetric mirror. These analytic expressions will be useful in mirror design by providing a straightforward way to compare the trade-offs between asymmetric and symmetric mirror designs.

Theory of reflectivity of an asymmetric mirror.

By using the concept of transfer matrices and Bloch waves, we have derived a set of equations that provide insight into the operation of asymmetric Bragg reflectors that have been demonstrated to be useful in achieving high reflectivities in strained-material systems. These equations will be useful in the design of asymmetric mirrors and can be used to compare the trade-offs between the conventional, symmetric (quarter-wavelength), and asymmetric mirrors.

Excessive carbohydrate intake in pregnancy and neonatal obesity: study in Cap Bon, Tunisia.

A high incidence (over 20%) of obesity was found in 250 neonates living in a rural area of Tunisia, by using weight and ponderal index per gestational age as the nutritional index. Maternal diabetes was probably excluded. Two surveys on nutritional habits--one on the general population and the other on pregnant women--showed a tendency to consume a high carbohydrate and low protein diet. The effect of a badly balanced maternal diet on the fetus is discussed.

Use of an "information collecting stamp" in maternal and child health centres.

PIP: Advantages of the use of a rubber stamp to aid in collecting client information at Maternal and Child Health care centers in Tunisia are discussed. The stamp's use insures standardised data collection on pertinent points related to the health of mothers and infants who are regular clinic users. The child's card is stamped at each visit and information is then filled at each designated area. These are: breast feeding, artificial feeding, whether the child is fed with his family, age of weaning, whether the mother is pregnant and how many months, and whether she has had a tetanus shot. The Nurse can make recommendations on feeding with the use of a specific chart for adequate nourishment for a Tunisian infant up to 11 months of age. Whether the mother has followed previous recommendations is determined by comparing current answers with those noted down previously. Staff errors are also easily pinpointed by the discrepancies they make in noting down various information. Staff education can then be geared accordingly. The resulting well standardised collection of data will be useful in specific studies on the clinics and the area.^ieng