Evaluation of a new arterial pressure-based cardiac output device requiring no external calibration

Background

Experience suggests that patients with alcohol and other drug use disorders (AOD) are commonly cared for in our intensive care units (ICU's) and require more sedation. We sought to determine the impact of AOD on sedation requirement and mechanical ventilation (MV) duration.

Methods

Retrospective review of randomly selected records of adult patients undergoing MV in the medical ICU. Diagnoses of AOD were identified using strict criteria in Diagnostic and Statistical Manual of Mental Disorders, and through review of medical records and toxicology results.

Results

Of the 70 MV patients reviewed, 27 had AOD (39%). Implicated substances were alcohol in 22 patients, cocaine in 5, heroin in 2, opioids in 2, marijuana in 2. There was no difference between AOD and non-AOD patients in age, race, or reason for MV, but patients with AOD were more likely to be male (21 versus 15, p < 0.0001) and had a lower mean Acute Physiology and Chronic Health Evaluation II (22 versus 26, p = 0.048). While AOD patients received more lorazepam equivalents (0.5 versus 0.2 mg/kg.day, p = 0.004), morphine equivalents (0.5 versus 0.1 mg/kg.day, p = 0.03) and longer duration of infusions (16 versus 10 hours/day. medication, p = 0.002), they had similar sedation levels (Richmond Agitation-Sedation Scale (RASS) -2 versus -2, p = 0.83), incidence of agitation (RASS ≥ 3: 3.0% versus 2.4% of observations, p = 0.33), and duration of MV (3.6 versus 3.9 days, p = 0.89) as those without AOD.

Conclusion

The prevalence of AOD among medical ICU patients undergoing MV is high. Patients with AOD receive higher doses of sedation than their non-AOD counterparts to achieve similar RASS scores but do not undergo longer duration of MV.     

Susceptibility of β1 Na+-K+ pump subunit to glutathionylation and oxidative inhibition depends on conformational state of pump

Glutathionylation of cysteine 46 of the β1 subunit of the Na(+)-K(+) pump causes pump inhibition. However, the crystal structure, known in a state analogous to an E2·2K(+)·P(i) configuration, indicates that the side chain of cysteine 46 is exposed to the lipid bulk phase of the membrane and not expected to be accessible to the cytosolic glutathione. We have examined whether glutathionylation depends on the conformational changes in the Na(+)-K(+) pump cycle as described by the Albers-Post scheme. We measured β1 subunit glutathionylation and function of Na(+)-K(+)-ATPase in membrane fragments and in ventricular myocytes. Signals for glutathionylation in Na(+)-K(+)-ATPase-enriched membrane fragments suspended in solutions that preferentially induce E1ATP and E1Na(3) conformations were much larger than signals in solutions that induce the E2 conformation. Ouabain further reduced glutathionylation in E2 and eliminated an increase seen with exposure to the oxidant peroxynitrite (ONOO(-)). Inhibition of Na(+)-K(+)-ATPase activity after exposure to ONOO(-) was greater when the enzyme had been in the E1Na(3) than the E2 conformation. We exposed myocytes to different extracellular K(+) concentrations to vary the membrane potential and hence voltage-dependent conformational poise. K(+) concentrations expected to shift the poise toward E2 species reduced glutathionylation, and ouabain eliminated a ONOO(-)-induced increase. Angiotensin II-induced NADPH oxidase-dependent Na(+)-K(+) pump inhibition was eliminated by conditions expected to shift the poise toward the E2 species. We conclude that susceptibility of the β1 subunit to glutathionylation depends on the conformational poise of the Na(+)-K(+) pump.     

Ecophysiology and ultrastructure of Ancylonema nordenski?ldii (Zygnematales, Streptophyta), causing brown ice on glaciers in Svalbard (high arctic)

At the arctic archipelago of Svalbard, bare glacier surfaces are populated by microalgae like Ancylonema nordenski?ldii (Zygnematales, Streptophyta). The resulting blooms cause, due to a vacuolar pigmentation, brownish colourations of the glacier surface. This freshwater ice alga has been described from several polar and alpine glaciers; however, these reports lacked data about the ecophysiology or ultrastructure. Considering the harsh environmental conditions of the exceptional habitat, such as permanently low temperatures, exposure to high irradiation or a short vegetation period, the aim of this study was to elucidate cellular adaptations of A. nordenski?ldii. Thus, samples were collected at two glaciers in Spitsbergen. The cytoarchitecture of the cylindrical cells, which are arranged in unbranched filaments, demonstrates active cells with Golgi bodies, mitochondria and rough endoplasmic reticulum close to the nucleus when investigated by transmission electron microscopy (TEM). The cell walls are pore less and only 90 nm thin. A. nordenski?ldii only sporadically produces oblong zygotes when two filaments conjugate. The most remarkable cytological feature is peripheral brownish vacuoles, appearing osmiophil and electron dense by TEM. Aqueous extracts of this pigmentation show a broad absorption in the visible light and in the UV. Consequently, a protection against excessive irradiation is provided. Photosynthesis measurements performed at different temperatures and light levels indicate that the metabolism is adapted to temperatures close to the freezing point as well as to high light conditions. Therefore, A. nordenski?ldii can be regarded as metabolically and cytological well adapted to live on glaciers.     

Regulation of the activities of the mammalian transglutaminase family of enzymes

Mammalian transglutaminases catalyze post‐translational modifications of glutamine residues on proteins and peptides through transamidation or deamidation reactions. Their catalytic mechanism resembles that of cysteine proteases. In virtually every case, their enzymatic activity is modulated by elaborate strategies including controlled gene expression, allostery, covalent modification, and proteolysis. In this review, we focus on our current knowledge of post‐translational regulation of transglutaminase activity by physiological as well as synthetic allosteric agents. Our discussion will primarily focus on transglutaminase 2, but will also compare and contrast its regulation with Factor XIIIa as well as transglutaminases 1 and 3. Potential structure–function relationships of known mutations in human transglutaminases are analyzed.     

Effects of spinosad, spinosad bait, and chloronicotinyl insecticides on mortality and control of adult and larval western cherry fruit fly (Diptera: Tephritidae)

Effects of spinosad, spinosad bait, and the chloronicotinyl insecticides imidacloprid and thiacloprid on mortality of the adults and larvae of western cherry fruit fly, Rhagoletis indifferens Curran (Diptera: Tephritidae), were determined in the laboratory and the field. Spinosad and spinosad bait caused higher adult mortality than imidacloprid, which caused higher mortality than thiacloprid. Only spinosad bait prevented oviposition. All materials were more toxic to adults when ingested than when topically applied. Spinosad bait had the greatest residual toxicity on leaves, killing 100% of adults when aged for 14 d in the field. When materials were sprayed on infested cherries, numbers of live larvae in fruit after 8 d were lower in imidacloprid and thiacloprid than in spinosad and spinosad bait treatments, which did not differ from the control, but all materials reduced larval emergence over 30 d. In the field, spinosad and spinosad bait were as effective in suppressing larval infestations as azinphos-methyl and carbaryl, whereas imidacloprid was effective in most cases and thiacloprid was generally less effective than azinphos-methyl and carbaryl. Overall, results in the laboratory and field show that spinosad and chloronicotinyl insecticides differed significantly in their effectiveness against adults and larvae of R. indifferens but that spinosad, spinosad bait, and imidacloprid seem to be acceptable substitutes for organophosphate and carbamate insecticides for controlling this fruit fly.     

Molecular flexibility in protein-DNA interactions

In living cells protein-DNA interactions are fundamental processes. Here, we compare the 3D structures of several DNA-binding proteins frequently determined with and without attached DNA. We studied the global structure (backbone-traces) as well as the local structure (binding sites) by comparing pair-wise the related atoms. The DNA-interaction sites of uncomplexed proteins show conspicuously high local structural flexibility. Binding to DNA results in specific local conformations, which are clearly distinct from the unbound states. The adaptation of the protein's binding site to DNA can never be described by the lock and key model but in all cases by the induced fit model. Conformational changes in the seven protein backbone traces take place in different ways. Two of them dock onto DNA without a significant change, while the other five proteins are characterized by a backbone conformation change caused by DNA docking. In the case of three proteins of the latter group the DNA-complexed conformation also occurs in a few uncomplexed structures. This behavior can be described by a conformational ensemble, which is narrowed down by DNA docking until only one single DNA-complexed conformation occurs. Different docking models are discussed and each of the seven proteins is assigned to one of them.     

Candida albicans SET1 encodes a histone 3 lysine 4 methyltransferase that contributes to the pathogenesis of invasive candidiasis

Candida albicans causes diverse mucosal and systemic diseases. Although this versatility likely depends upon carefully co-ordinated gene expression, epigenetic regulation in C. albicans remains poorly characterized. Screening a genomic expression library, we identified C. albicans Set1p as an immunogenic protein with homology to a lysine histone methyltransferase of Saccharomyces cerevisiae. In this study, we demonstrated that total immunoglobulin, IgG and IgM titers against a unique Set1p N-terminal fragment were significantly higher among patients with disseminated candidiasis (DC) or oropharyngeal candidiasis than controls. Disruption of SET1 resulted in complete loss of methylation of histone 3 at lysine residue 4, hyperfilamentous growth under embedded conditions, less negative cell surface charges and diminished adherence to epithelial cells, effects that were reversed upon gene re-insertion at a disrupted locus. During murine DC, the null mutant was associated with prolonged survival and lower tissue burdens. Taken together, our findings suggest that SET1 regulates multiple processes important to the pathogenesis of candidiasis. The Set1p N-terminal fragment does not exhibit significant homology to eukaryotic or microbial proteins, and might represent a novel therapeutic, preventive or diagnostic target.     

Synthesis and SAR of pyrrolotriazine-4-one based Eg5 inhibitors

Synthesis and SAR of substituted pyrrolotriazine-4-one analogues as Eg5 inhibitors are described. Many of these analogues displayed potent inhibitory activities in the Eg5 ATPase and A2780 cell proliferation assays. In addition, pyrrolotriazine-4-one analogue 26 demonstrated in vivo efficacy in an iv P388 murine leukemia model. Both NMR and X-ray crystallographic studies revealed that these analogues bind to an allosteric site on the Eg5 protein.     

Response of arctic snow and permafrost algae to high light and nitrogen stress by changes in pigment composition and applied aspects for biotechnology

Ten algal strains from snow and permafrost substrates were tested for their ability to produce secondary carotenoids and α-tocopherol in response to high light and decreased nitrogen levels. The Culture Collection of Cryophilic Algae at Fraunhofer IBMT in Potsdam served as the bioresource for this study. Eight of the strains belong to the Chlorophyceae and two strains are affiliated to the Trebouxiophyceae . While under low light, all 10 strains produced the normal spectrum of primary pigments known to be present in Chlorophyta, only the eight chlorophyceaen strains were able to synthesize secondary carotenoids under stress conditions, namely canthaxanthin, echinenone and astaxanthin; seven of them were also able to synthesize minor amounts of adonixanthin and an unidentified hydroxyechinenone. The two trebouxiophyceaen species of Raphidonema exhibited an unusually high pool of primary xanthophyll cycle pigments, possibly serving as a buffering reservoir against excessive irradiation. They also proved to be good α-tocopherol producers, which might also support the deactivation of reactive oxygen species. This study showed that some strains might be interesting novel candidates for biotechnological applications. Cold-adapted, snow and permafrost algae might serve as valuable production strains still exhibiting acceptable growth rates during the cold season in temperate regions.