Correction to: In vitro cellular and proteome assays identify Wnt pathway and CDKN2A-regulated senescence affected in mesenchymal stem cells from mice after a chronic LD gamma irradiation in utero

Radiation and Environmental Biophysics -     

Reconstitution and fusogenic properties of Sendai virus envelopes

Sendai virus membranes were reconstituted by detergent dialysis, using the non-ionic detergents Triton X-100 and octyl glucoside. Membrane reassembly was determined by measuring the surface-density-dependent efficiency of resonance energy transfer between two fluorescent phospholipid analogues, which were co-reconstituted with the viral envelopes. The functional incorporation of the viral proteins was established by monitoring the ability of the reconstitution products to fuse with erythrocyte membranes, utilizing assays based on either resonance energy transfer or on relief of fluorescence selfquenching. The persistent adherence of residual Triton X-100 with the reconstituted membrane was revealed by an artificial detergent-effect on the resonance energy transfer efficiency and the occurrence of hemolysis of human erythrocytes under conditions where fusion does not occur. Properly reconstituted Sendai virus envelopes were obtained with octyl glucoside. The fusion activity of the viral envelopes was dependent on the initial concentration of octyl glucoside used to disrupt the virus and the rate of detergent removal. Rapid removal of detergent by dialysis against large volumes of dialysis buffer (ratio 1:850) or by gel filtration produced reconstituted membranes capable of inducing hemagglutination but significant fusion activity was not detected. By decreasing the volume ratio of dialysate versus dialysis buffer to 1:250 or 1:25, fusogenic viral envelopes were obtained. The initial fusion kinetics of the reconstituted viral membrane and the parent virus were different in that both the onset and the initial rate of fusion of the reconstituted membranes were faster, whereas the extents to which both particles eventually fused with the target membrane were similar. The differences in the initial fusion kinetics lead us to suggest that the details of the fusion mechanism between Sendai virus and the target membrane involve factors other than the mere presence of glycoproteins F and HN in the viral bilayer. Finally, the results also indicate that determination of the viral fusion activity in a direct manner, rather than by an indirect assay, such as hemolysis, is imperative for a proper evaluation of the functional properties retained upon viral reconstitution.     

Nuclear corrosion monitoring—

Nuclear corrosion technique has been developed for the assay of various heavy metals released through corrosion and abrasion into electrolytes from various biomaterials like amalgams, chromium— cobalt and gold alloys, steel, and titanium. Application of the technique in measurement of selective release rates under static or dynamic conditions, i.e., during cyclic loading, is discussed. The elements chromium, cobalt, copper, gold, iron, mercury, molybdenum, silver, titanium, and zinc have been quantitatively assessed. In vivo corrosion measurements are further included. By combining the present nuclear tracer technique with ESCA technique, knowledge about reaction mechanisms occurring at the interface solid/liquid is obtained. Exposure of humans to various heavy metals from biomaterials, e.g., dental materials, can be estimated using the NCM technique. The technique also has a potential for selective release measurements of several nuclides possessing suitable radioanalytical properties from other types of alloys immersed in various liquid environments.     

Transducer-binding and transducer-mutations modulate photoactive-site-deprotonation in sensory rhodopsin I.

Sensory rhodopsin I (SRI) is a seven-transmembrane helix retinylidene protein that mediates color-sensitive phototaxis responses through its bound transducer HtrI in the archaeon Halobacterium salinarum. Deprotonation of the Schiff base attachment site of the chromophore accompanies formation of the SRI signaling state, S(373). We measured the rate of laser flash-induced S(373) formation in the presence and absence of HtrI, and the effects of mutations in SRI or HtrI on the kinetics of this process. In the absence of HtrI, deprotonation occurs rapidly (halftime 10 micros) if the proton acceptor Asp76 is ionized (pK(a) = approximately 7), and only very slowly (halftime > 10 ms) when Asp76 is protonated. Transducer-binding, although it increases the pK(a) of Asp76 so that it is protonated throughout the range of pH studied, results in a first order, pH-independent rate of S(373) formation of approximately 300 micros. Therefore, the complexation of HtrI facilitates the proton-transfer reaction, increasing the rate approximately 50-fold at pH6. Arrhenius analysis shows that HtrI-binding accelerates the reaction primarily by an entropic effect, suggesting HtrI constrains the SRI molecule in the complex. Function-perturbing mutations in SRI and HtrI also alter the rate of S(373) formation and the lambda(max) of the parent state as assessed by laser flash-induced kinetic difference spectroscopy, and shifts to longer wavelength are correlated with slower deprotonation. The data indicate that HtrI affects electrostatic interactions of the protonated Schiff base and not only receives the signal from SRI but also optimizes the photochemical reaction process for SRI signaling.     

Responses in pigmentation and anti-oxidant expression in Arctic Daphnia along gradients of DOC and UV exposure

Responses in carapace melanization and expression of the majoranti-oxidant catalase (CAT) and glutathione transferase (GST)in Arctic Daphnia were assessed in enclosures along a gradientof dissolved organic carbon (DOC). This gradient was createdby adding freeze-dried humic matter to 2 m³ UV-transparent enclosures,yielding final nominal concentrations of 1, 2.5, 5 and 10 mgC l^-1. The UV attenuation was strongly affected by additionsof DOC, and attenuation coefficients at 320 nm increased from3.0 in the control to approximately 3.5 and 11.0 m^-1 in the1 and 10 mg DOC treatments respectively. Most Daphnia showedpronounced carapace melanization, and the absorbance of short-waveradiation through the carapace was strongly related to the degreeof melanization. Nevertheless, the different UV climate in theenclosures did not cause any short-term adaptation in Daphniapigmentation over a 3 week period. The levels of CAT and GSTwere assessed over time in the control and at 10 mg DOC. Theseenzymes displayed opposite patterns, with somewhat lower activitiesof CAT at low DOC (control) relative to 10 mg DOC, while theopposite was found for GST. There was also a significant negativecorrelation between CAT and solar irradiation for GST in bothbags, while no effects were found for GST.     

Locomotor Activity Patterns in Ilybius fenestratus (Fabricius) (Coleoptera,Dytiscidae) in a Central Norwegian Lake,with Evidence for Breeding-Induced Arrhythmicity

The diel activity of Ilybius fenestratus was studied in Målsj?en, a lake in S?r-Tr?ndelag, Central Norway (63°14'N, 10°26'E), during 1971–1972. Adults and larvae were sampled in activity traps every week during March–October and every 2–3 weeks during November–February. The traps were emptied every 2 hrs during a 24- hr period. The diel locomotor activity of I. fenestratus was clearly nocturnal, except in the second half of July, for the males also in the first half of July, when surprisingly the population showed an arrhythmic activity pattern. The arrhythmicity is connected with the mating and egg-laying period of the species. The larvae were also nocturnal in the autumn, but showed arrhythmicity under the ice during the winter.     

Bio-catalyzed coloration of cellulose fibers

Cotton fabrics were dyed with dyes generated in situ by laccase-catalyzed oxidative coupling of the colorless 2,5-diaminobenzenesulfonic acid (2,5-DABSA) and 1-hydroxyphenol (catechol). The enzymatic oxidation of the dye intermediates led to cross-coupling reaction products when the reaction was conducted with an excess of catechol. At least fourfold excess of catechol was necessary to achieve satisfactory dye fixation on cotton. Formation of the same colored product using either an equimolar ratio of the reagents or tenfold excess of catechol was observed. Most probably, homo-molecular reactions predominate over the cross-coupling at equimolar ratio of the precursors, while with an excess of catechol, the cross-coupling occurs in higher yield. The reaction was followed using UV-Vis spectroscopy, HPLC, FTIR and MALDI-TOF MS. A reaction pathway for laccase-induced cross-coupling of catechol and 2,5-DABSA yielding a major colored product was proposed.