Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes

Aims

The diverse physiological functions of histamine are mediated through distinct histamine receptors. In this study we investigated the role of H₂R and H₄R in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood.

Main methods

Changes in reactive oxygen species (ROS) production by whole blood phagocytes after treatment with histamine, H₄R agonists (4-methylhistamine, VUF8430), H₂R agonist (dimaprit) and their combinations with H₄R antagonist (JNJ10191584) and H₂R antagonist (ranitidine) were determined using the chemiluminescence (CL) assay. To exclude the direct scavenging effects of the studied compounds on the CL response, the antioxidant properties of all compounds were measured using several methods (TRAP, ORAC, and luminol–HRP–H₂O₂ based CL).

Key findings

Histamine, 4-methylhistamine, VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner. On the other hand, only VUF8430 was able to inhibit PMA-activated whole blood CL. Ranitidine, but not JNJ10191584, completely reduced the effects of histamine, 4-methylhistamine and dimaprit. The direct scavenging ability of tested compounds was negligible.

Significance

Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by H₂R. Our results also suggest that H₄R agonists in concentrations higher than 10^− 6 M may also influence ROS production via binding to H₂R.     

The effect of electron acceptors on the phytochrome-controlled germination of Paulownia tomentosa seeds

The effect of substances with different redox potentials in the phytochrome-controlled germination of Paulmtnia tomentosa seeds was examined. Up to 25% of water-imbibed seeds germinated upon irradiation with 5 min red light The seeds irradiated with 5 min red light and incubated in substances with redox potentials equal or higher than E'₀= 360 mV [potassium nitrate, potassium hexacyanoferrate (III) or potassium hexachloroiridate (IV)] and sodium nitroprusside germinated up to 80%. The optimal concentration was between 1 and 10 mM. Other electron acceptors such as 2.6-dichlorophenol-indoplhenol, phenazine methosulfate and methylene blue failed to produce and effect. The germination-promoting effect was obtained when the substances were supplied to the incubation medium from the onset of imbibition or at the onset of irradiation. Delay of application of these chemicals until after the red light treatment diminished their effectiveness and the effect was lost after 48 h. The escape from far-red reversibility was evident in the presence of substances which stimulated germination. The results presented support the view that phytochrome-broken dormancy of Paulownia tomentosa seeds is closely connected with redox changes.     

Marburg variant multiple sclerosis - a case report

In this case report we describe the case of a 24 year-old female with a fulminant demyelinating disease of white matter. Disease progression was most probably consistent with the Marburg variant (malignant form) of multiple sclerosis with rapid deterioration of the patient's clinical condition, including bulbar symptoms and epileptic paroxysms and ending with persistent coma and tetraparesis, over the course of 6 months from first symptoms. Repeated Magnetic Resonance Imaging (MRI) examination showed progression of multiple demyelinating lesions culminating in a contiguous focal disorder of the white matter extending both supratentorially and infratentorially. The serial MRI changes closely mapped the deterioration in the patients clinical status. Our patient showed no response to repeated pulse corticotherapy, administration of intravenous immunoglobulins, serial plasmapheresis, and combined high-dose pulse immunosuppression (specify what was used here) and mitoxantrone.     

Activation-dependent descending reflex evacuation motority of anal canal in rat model

The evacuative motor responses of the anal canal and recto-anal reflexes during defecation were studied in an isolated rat recto-anal model preparation using (i) partitioned organ bath, (ii) electrical stimulation, (iii) balloon distension and (iv) morphological techniques. Electrical field stimulation applied to the anal canal or to the distal part of the rectum elicited tetrodotoxin (10(-7) M)-sensitive frequency-dependent local or descending contractions of the anal canal and the local responses were bigger in amplitude (14.9 ± 1.35 mN) than the descending contractions (5.3 ± 0.7 mN at frequency of 5 Hz, p < 0.05). The balloon-induced distension of the distal rectum evoked descending responses of the anal canal consisting of a short contraction (1.50 ± 0.18 mN) followed by deep relaxation (3.12 ± 0.34 mN). In the presence of atropine (3 x 10(-7) M) the electrically-elicited (5 Hz) local or descending contractions of the anal canal were suppressed and a relaxation revealed. The initial contraction component of the distension-induced response was decreased while the relaxation was not changed. During atropine treatment, spantide (10(-7) M) lowered even more the contractile component of the anal canal response. NG-nitro-L-arginine (5 x 10(-4) M) enhanced the contraction, prevented the atropine-dependent relaxation of the electrically-elicited response and inhibited the distension-induced relaxation. L-Arginine (5 x 10(-4) M) suppressed the contraction and extended the relaxation. ChAT-, substance P- and NADPH-diaphorase-positive perikarya and nerve fibers were observed in myenteric ganglia of the anal canal. The results suggest activation-dependent descending reflex motority of the anal canal involving electrical stimulation-displayed cholinergic and tachykininergic and distension manifested nitrergic neuro-muscular communications.     

Secondary alcohol dehydrogenase catalyzes the reduction of exogenous acetone to 2-propanol in Trichomonas vaginalis

Secondary alcohols such as 2-propanol are readily produced by various anaerobic bacteria that possess secondary alcohol dehydrogenase (S-ADH), although production of 2-propanol is rare in eukaryotes. Specific bacterial-type S-ADH has been identified in a few unicellular eukaryotes, but its function is not known and the production of secondary alcohols has not been studied. We purified and characterized S-ADH from the human pathogen Trichomonas?vaginalis. The kinetic properties and thermostability of T.?vaginalis S-ADH were comparable with bacterial orthologues. The substantial activity of S-ADH in the parasite's cytosol was surprising, because only low amounts of ethanol and trace amounts of secondary alcohols were detected as metabolic end products. However, S-ADH provided the parasite with a high capacity to scavenge and reduce external acetone to 2-propanol. To maintain redox balance, the demand for reducing power to metabolize external acetone was compensated for by decreased cytosolic reduction of pyruvate to lactate and by hydrogenosomal metabolism of pyruvate. We speculate that hydrogen might be utilized to maintain cytosolic reducing power. The high activity of Tv-S-ADH together with the ability of T.?vaginalis to modulate the metabolic fluxes indicate efficacious metabolic responsiveness that could be advantageous for rapid adaptation of the parasite to changes in the host environment.     

Role of Us9 Phosphorylation in Axonal Sorting and Anterograde Transport of Pseudorabies Virus

Alphaherpes viruses, such as pseudorabies virus (PRV), undergo anterograde transport in neuronal axons to facilitate anterograde spread within hosts. Axonal sorting and anterograde transport of virions is dependent on the viral membrane protein Us9, which interacts with the host motor protein Kif1A to direct transport. Us9-Kif1A interactions are necessary but not sufficient for these processes, indicating that additional cofactors or post-translational modifications are needed. In this study, we characterized two conserved serine phosphorylation sites (S51 and S53) in the PRV Us9 protein that are necessary for anterograde spread in vivo. We assessed the subcellular localization of phospho-Us9 subspecies during infection of neurons and found that the phospho-form is detectable on the majority, but not all, of axonal vesicles containing Us9 protein. In biochemical assays, phospho-Us9 was enriched in lipid raft membrane microdomains, though Us9 phosphorylation did not require prior lipid raft association. During infections of chambered neuronal cultures, we observed only a modest reduction in anterograde spread capacity for diserine mutant Us9, and no defect for monoserine mutants. Conversely, mutation of the kinase recognition sequence residues adjacent to the phosphorylation sites completely abrogated anterograde spread. In live-cell imaging analyses, anterograde transport of virions was reduced during infection with a recombinant PRV strain expressing GFP-tagged diserine mutant Us9. Phosphorylation was not required for Us9-Kif1A interaction, suggesting that Us9-Kif1A binding is a distinct step from the activation and/or stabilization of the transport complex. Taken together, our findings indicate that, while not essential, Us9 phosphorylation enhances Us9-Kif1A-based transport of virions in axons to modulate the overall efficiency of long-distance anterograde spread of infection.     

The Maghreb – one more important biodiversity hot spot for tiger beetle fauna (Coleoptera,Carabidae, Cicindelinae) in the Mediterranean region

The tiger beetle fauna of the Maghreb region is one of the richest in the Palaearctic, including 22 species and 5 subspecies and 19% of all Palaearctic species of Cicindelinae. Assembled to their chorotypes, the Maghreb tiger beetles fall into eight different groups that include Maghreb endemics (26% of fauna), Mediterranean (7%), West Mediterranean (40%), North African (4%), Mediterranean-Westturanian (4%), West Palaearctic (4%), Afrotropico-Indo-Mediterranean (4%), and Saharian (11%) species. The Mediterranean Sclerophyl and Atlas Steppe are the Maghreb biogeographical provinces with the highest species richness, while the Sahara Desert has the lowest Cicindelinae diversity. Twenty-five cicindelid species and subspecies (93% of Maghreb fauna) are restricted to only one or two habitat types in lowland areas. Only Calomera littoralis littoralis and Lophyra flexuosa flexuosa are recognized as eurytopic species and occur in three types of habitat. The highest tiger beetle diversity characterizes salt marshes and river banks (in both cases 11 species and subspecies or 41% of Maghreb fauna). Approximately 85% of all Maghreb tiger beetle species and subspecies are found in habitats potentially endangered by human activity.     

An InCytes from MBC Selection: Importin β Regulates the Seeding of Chromatin with Initiation Sites for Nuclear Pore Assembly

The nuclear envelope of higher eukaryotic cells reforms at the exit from mitosis, in concert with the assembly of nuclear pore complexes (NPCs). The first step in postmitotic NPC assembly involves the “seeding” of chromatin with ELYS and the Nup107-160 complex. Subsequent steps in the assembly process are poorly understood and different mechanistic models have been proposed to explain the formation of the full supramolecular structure. Here, we show that the initial step of chromatin seeding is negatively regulated by importin β. Direct imaging of the chromatin attachment sites reveals single sites situated predominantly on the highest substructures of chromatin surface and lacking any sign of annular structures or oligomerized pre-NPCs. Surprisingly, the inhibition by importin β is only partially reversed by RanGTP. Importin β forms a high-molecular-weight complex with both ELYS and the Nup107-160 complex in cytosol. We suggest that initiation sites for NPC assembly contain single copies of chromatin-bound ELYS/Nup107-160 and that the lateral oligomerization of these subunits depends on the recruitment of membrane components. We predict that additional regulators, besides importin β and Ran, may be involved in coordinating the initial seeding of chromatin with subsequent steps in the NPC assembly pathway.