Tularemia progression accompanied with oxidative stress and antioxidant alteration in spleen and liver of BALB/c mice

Francisella tularensis is the causative agent of tularemia. It is an intracellular pathogen with the ability to survive within phagosomes and induce pyroptotic cell death. In this study, we attempted to prove whether oxidative imbalance plays a significant role in tularemia pathogenesis. In our experimental model, we subcutaneously infected female BALB/c mice (dose 10(5) CFU of F. tularensis LVS). Liver, spleen, and blood were collected from mice at regular intervals from days 1-15 after infection. The bacterial burden was assessed by a cultivation test. The burden was unchanging from the 2(nd) to 6(th) day after infection. The bacterial burden corresponded to the plasmatic level of IFN-γ, IL-6, and liver malondialdehyde. After the phase of acute bacteraemia and the innate immunity reaction, the levels of reduced glutathione and total low molecular weight antioxidants decreased significantly and the activity of caspase-3 increased in the liver. The level of reduced glutathione decreased to 25% of the original level, and the total level of low molecular weight antioxidants was less than 50% of the initial amount. The demonstrated effects of tularemia-induced pathology had a more extensive impact on the liver than on the spleen.     

Amniotic fluid cathelicidin in PPROM pregnancies: from proteomic discovery to assessing its potential in inflammatory complications diagnosis

Background

Preterm prelabor rupture of membranes (PPROM) complicated by microbial invasion of the amniotic cavity (MIAC) leading to histological chorioamnionitis (HCA) significantly impacts perinatal morbidity. Unfortunately, no well-established tool for identifying PPROM patients threatened by these disorders is available.

Methodology/Principal Findings

We performed an unbiased exploratory analysis of amniotic fluid proteome changes due to MIAC and HCA. From among the top five proteins that showed the most profound and significant change, we sought to confirm results concerning cathelicidin ({type:entrez-protein,attrs:{text:P49913,term_id:1706745,term_text:P49913}}P49913, CAMP_HUMAN), since an ELISA kit was readily available for this protein. In our exploratory proteomic study, cathelicidin showed a ∼6-fold higher concentration in PPROM patients with confirmed MIAC and HCA. We verified significantly higher levels of cathelicidin in exploratory samples (women without both MIAC and HCA: median 1.4 ng/ml; women with both conditions confirmed: median 3.6 ng/ml; p = 0.0003). A prospective replication cohort was used for independent validation and for assessment of cathelicidin potential to stratify women with MIAC leading to HCA from women in whom at least one of these conditions was ruled out. We confirmed the association of higher amniotic fluid cathelicidin levels with MIAC leading to HCA (the presence of both MIAC and HCA: median 3.1 ng/ml; other women: median 1.4 ng/ml; p<0.0001). A cathelicidin concentration of 4.0 ng/ml was found to be the best cut-off point for identifying PPROM women with both MIAC and HCA. When tested on the validation cohort, a sensitivity of 48%, a specificity of 90%, a likelihood ratio of 5.0, and an area under receiver-operating characteristic curve of 71% were achieved for identification of women with MIAC leading to HCA.

Conclusions

Our multi-stage study suggests cathelicidin as a candidate marker that should be considered for a panel of amniotic fluid proteins permitting identification of PPROM women with MIAC leading to HCA.     

Mechanisms of salt and water transport in proximal tubule as measured by split drop

A mathematical analysis of split drop experiments indicates that the isosmotic flow assumption is naturally embedded in the system equations. The analysis is applied to the experimental data reported by Maude (1970), who used a polyethylene glycol (PEG 1000) and sodium chloride perfusate in rat proximal tubule. In addition to a value of the permeability coefficient of the slowly permeating species (PEG 1000) which is in accord with Maude's findings, upper limits for the values of sodium and water permeability coefficients are calculated. In particular, it was found that the sodium permeability coefficient is, at most, three times larger than that of PEG 1000. A good fit to the data is provided by a passive transport model.     

Local versus regional species richness in tropical insects: one lowland site compared with the island of New Guinea

1. The overlap in species composition of Cercopoidea (Aphrophoridae, Cercopidae, and Machaerotidae), Flatidae, and Ricaniidae between two data sets, an almost exhaustive census from 13 Ficus species and a sample from diverse vegetation in the same area, led to the estimate for local species richness of 111 (SE 11.5) species (45 species of Cercopoidea, 36 species of Flatidae, and 30 species of Ricaniidae) at a lowland rainforest site in New Guinea. 2. Samples restricted to 13 species of Ficus contained 66 species, i.e. 59% of the estimated local species richness. This high proportion probably results from the high proportion of polyphagous and tourist (transient) species in the Cercopoidea, Flatidae, and Ricaniidae. 3. The two largest museum collections of New Guinean insects contained 327 species of Cercopoidea from New Guinea, including 23 of the 34 species collected in the field samples. This overlap led to the estimate of 483 (SE 97.2) species of Cercopoidea present in New Guinea. 4. The species found in the field samples were also 2.6 times more likely to be found in the museum collection than other species. This sampling bias can be due to a positive correlation between species local abundance and geographic distribution and/or similar patterns of species abundance at different sites. 5. The estimate of species richness of Cercopoidea in New Guinea increased to 1222 species when corrected for this sampling bias. Thus, only 4% of the New Guinean species were present locally, in the study area. Such high beta diversity is probably a consequence of the exceptional habitat and vegetation diversity in New Guinea, as well as its complex geological history.     

Isolation and biochemical characterization of two soluble iron(III) reductases from Paracoccus denitrificans.

Two soluble enzymes (FerA and FerB) catalyzing the reduction of a number of iron(III) complexes by NADH, were purified to near homogeneity from the aerobically grown iron-limited culture of Paracoccus denitrificans using a combination of anion-exchange chromatography (Sepharose Q), chromatofocusing (Mono P), and gel permeation chromatography (Superose 12). FerA is a monomer with a molecular mass of 19 kDa, whereas FerB exhibited a molecular mass of about 55 kDa and consists of probably two identical subunits. FerA can be classified as an NADH:flavin oxidoreductase with a sequential reaction mechanism. It requires the addition of FMN or riboflavin for activity on Fe(III) substrates. In these reactions, the apparent substrate specificity of FerA seems to stem exclusively from different chemical reactivities of Fe(III) compounds with the free reduced flavin produced by the enzyme. Observations on reducibility of Fe(III) chelated by vicinal dihydroxy ligands support the view that FerA takes part in releasing iron from the catechol type siderophores synthesized by P. denitrificans. Contrary to FerA, the purified FerB contains a noncovalently bound redox-active FAD coenzyme, can utilize NADPH in place of NADH, does not reduce free FMN at an appreciable rate, and gives a ping-pong type kinetic pattern with NADH and Fe(III)-nitrilotriacetate as substrates. FerB is able to reduce chromate, in agreement with the fact that its N-terminus bears a homology to the previously described chromate reductase from Pseudomonas putida. Besides this, it also readily reduces quinones like ubiquinone-0 (Q0) or unsubstituted p-benzoquinone.     

Effects of marijuana on testosterone in male subjects

Clinical studies have given contradictory reports on the effect of smoking marijuana on the plasma levels of testosterone in males. A reanalysis of existing data established that testosterone levels are depressed both after smoking one marijuana cigarette and after intravenous infusion of delta-9-tetrahydrocannabinol, a pharmacologically active component of marijuana. Simulation of the marijuana interaction, under the assumption that delta-9-tetrahydrocannabinol inhibits testosterone production or secretion, suggests a minimum of 24 hours are required for testosterone to return to pre-smoking levels. A series of clinical studies are specified to clarify the nature of the interaction.     

Transport across epithelia: A kinetic evaluation

Compartmental analysis of three models for solute transport across epithelial tissue is presented. The simplest model describes only one tissue compartment, the second incorporates the notion of a pore as a parallel pathway and the third model introduces a serial compartment corresponding to non-epithelial portions of the tissue. Experimental data were obtained, using a modified Ussing and Zerahn technique ((1951) Acta Physiol. Scand. 23, 110–127), for salicylate transport across rat jejunum in vitro and analyzed in terms of these three models. The conclusions based solely on the mathematical analysis of this rather simple experiment are: the tissue is not a homogeneous penetration barrier as often considered. Transport is not limited by unstirred layers either at the tissue surfaces or within the tissue itself. Salicylate is not passively transported. Parallel transport pathways do exist.     

Bimolecular systems: III. Catalytic systems

Amplification effect in the catalytic bimolecular systems is a consequence of the kinetic characteristic of the catalyst. Two types of the coefficient of amplification are defined. The applicability of these definitions is given by the type of the bimolecular system. In a simple example it is shown that the concept of amplification is meaningful in these systems. Furthermore, two rules, analogous to those for a coupling of amplifiers, are derived for the two basic modes of coupling of catalytic systems. Thus, in biological systems the catalytic reactions may be regarded as biologically effective amplifiers.