Biological time-dependent difference in effect of peroxynitrite demonstrated by the mouse hot plate pain model

We previously demonstrated the rhythmic pattern of L-arginine/nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) cascade in nociceptive processes. The coupled production of excess NO and superoxide leads to the formation of an unstable intermediate peroxynitrite, which is primarily responsible for NO-mediated toxicity. In the present study, we evaluated the biological time-dependent effects of exogenously administered peroxynitrite on nociceptive processes and peroxynitrite-induced changes in the analgesic effect of morphine using the mouse hot-plate pain model. Experiments were performed at four different times of day (1, 7, 13, and 19 hours after lights on, i.e., HALO) in mice of both sexes synchronized to a 12 h:12 h light-dark cycle. Animals were injected intraperitoneally (i.p.) with saline or 10 mg/kg morphine 30 min before and 0.001 mg/kg peroxynitrite 30 sec before hot-plate testing, respectively. The analgesic effect of morphine exhibited significant biological time-dependent differences in the thermally-induced algesia; whereas, administration of peroxynitrite alone exhibited either significant algesic or analgesic effect, depending on the circadian time of its injection. Concomitant administration of peroxynitrite and morphine reduced morphine-induced analgesia at three of the four different study time points. In conclusion, peroxynitrite displayed nociceptive and antinociceptive when administered alone according to the circadian time of treatment, while it diminished analgesic activity when administered in combination with morphine at certain biological times.     

The dynamics of homologous chromosome pairing during male Drosophila meiosis

BACKGROUND: Meiotic pairing is essential for the proper orientation of chromosomes at the metaphase plate and their subsequent disjunction during anaphase I. In male Drosophila melanogaster, meiosis occurs in the absence of recombination or a recognizable synaptonemal complex (SC). Due to limitations in available cytological techniques, the early stages of homologous chromosome pairing in male Drosophila have not been observed, and the mechanisms involved are poorly understood.RESULTS: Chromosome tagging with GFP-Lac repressor protein allowed us to track, for the first time, the behavior of meiotic chromosomes at high resolution, live, at all stages of male Drosophila meiosis. Homologous chromosomes pair throughout the euchromatic regions in spermatogonia and during the early phases of spermatocyte development. Extensive separation of homologs and sister chromatids along the chromosome arms occurs in mid-G2, several hours before the first meiotic division, and before the G2/M transition. Centromeres, on the other hand, show complex association patterns, with specific homolog pairing taking place in mid-G2. These changes in chromosome pairing parallel changes in large-scale chromosome organization.CONCLUSIONS: Our results suggest that widespread interactions along the euchromatin are required for the initiation, but not the maintenance, of meiotic pairing of autosomes in male Drosophila. We propose that heterochromatic associations, or chromatid entanglement, may be responsible for the maintenance of homolog association during late G2. Our data also suggest that the formation of chromosome territories in the spermatocyte nucleus may play an active role in ensuring the specificity of meiotic pairing in late prophase by disrupting interactions between nonhomologous chromosomes.     

Comparison of farm-level greenhouse gas emissions in transhumance and semi-intensive sheep production systems in continental rangelands

Despite their predominance worldwide, few studies have been conducted to look at the impact of sheep production systems relying on transhumance practices in arid and continental conditions, on farm-level greenhouse gas (GHG) emissions. Using Turkey as an example, this paper examines on farm-level GHG emissions calculated for two contrasting sheep production systems under arid and continental climate conditions. Production and management data were obtained through face-to-face interviews carried out on 10 transhumance and 15 semi-intensive meat sheep farms in Turkey. A total of seven GHG emission estimates were then calculated for each farm with the Agricultural Resource Efficiency Calculator (AgRECalc©) tool; i) total Carbon Dioxide (CO2) from energy use (kg CO2e), ii) total Carbon Dioxide equivalent (CO2e) from methane (kg CO2e), iii) total CO₂e from nitrous oxide (kg CO₂e), iv) whole farm and enterprise CO₂e emissions (kg CO₂e), v) net emission from land use (kg CO₂e), vi) whole farm CO₂e emissions per kg of farm output (kg CO₂e/kg output), vii) product CO₂e emissions (meat): kg CO₂e / kg live weight, and viii) farm output (kg of sheep). Multivariate analyses (using R software) were carried out to compare both farm types and their respective carbon emissions. The total farm output per ewe was lower in the transhumance farms (7.4 kg/ewe) than in the semi-intensive farms (7.7 kg/ewe). The kg CO₂e per kg of output was also lower for the transhumance farms (46.2 kg CO₂e) than for the semi-intensive ones (56.5 kg CO₂e). This trend was similar for the amount of CO₂e per kg of live weight produced (20.8 kg and 25.4 kg for the transhumance and the semi-intensive farms, respectively). Despite overall net emissions from land use being greater on average for the transhumance farms, once measured per hectare, they were found to be lower than those for the semi-intensive farms. This study provides a reference point for different sheep production systems’ GHG emission impact in continental rangelands in Turkey.     

Practical Image Restoration of Thick Biological Specimens Using Multiple Focus Levels in Transmission Electron Microscopy

Three-dimensional electron tomographic studies of thick specimens such as cellular organelles or supramolecular structures require accurate interpretations of transmission electron micrograph intensities. In addition to microscope lens aberrations, thick specimen imaging is complicated by additional distortions resulting from multiple elastic and inelastic scattering. Extensive analysis of the mechanism of image formation using electron energy-loss spectroscopy and imaging as well as exit wavefront reconstruction demonstrated that multiple scattering does not contribute to the coherent component of the exit wave (Hanet al.,1996, 1995). Although exit wavefront restored images showed enhanced contrast and resolution, that technique, which requires the collection of more than 30 images at different focus levels, is not practical for routine data collection in 3D electron tomography, where usually over 100 projection views are required for each reconstruction. Using a 0.7-μm-thick specimen imaged at 200 keV, the accuracy of reconstructions using small numbers of defocused images and a simple linear filter (Schiske, 1968) was assessed by comparison to the complete exit wave restoration. We demonstrate that only four optimal focus levels are required to effectively restore the coherent component (deviation 5.1%). By contrast, the optimal single image (zero defocus) shows a 25.5% deviation to the exit wave restoration. Two pairs of under- and over-defocus images should be taken: one pair at quite high defocus (>10 μm) to differentiate the coherent (single elastic scattering) from the incoherent (multiple elastic and inelastic scattering) components, and the second pair to optimize information content at the highest desired resolution (e.g., 5 μm for (2.5 nm)⁻¹resolution). We also propose a new interpretation of the restored amplitude and phase components where the specimen mass-density is proportional to the logarithm of the amplitude component and linearly related to the phase component. This approach should greatly facilitate the collection of high resolution tomographic data from thick samples.     

Performance evaluation and kinetic modeling of the start-up of a UASB reactor treating municipal wastewater at low temperature

A kinetic modeling-based study was carried out to evaluate the start-up performance of a 10-L up-flow anaerobic sludge blanket (UASB) reactor treating municipal wastewater under different organic and hydraulic loading conditions. The reactor was operated for 105 days (around 4 months) below 20 °C and with three different hydraulic retention times of 24, 12 and 5 h. Imposed volumetric organic loading rates (OLR) ranged from 0.57 (±0.05) to 11.78 (±0.85) kg TCOD/m³-day. Although relatively high incoming volumetric OLR values were employed to the system, the UASB reactor demonstrated a favorable performance on the anaerobic treatability of municipal wastewater, and no process failure was recorded in the start-up stage. On the basis of experimental results, the modified Stover–Kincannon model was successfully applied to define the start-up kinetics with a very high value of the correlation coefficient (R = 0.9729). Maximum substrate utilization rate constant and saturation constant of the modified Stover–Kincannon model were determined as U _max = 1.996 g/L-day and K _B = 1.536 g/L-day, respectively.     

Passive wireless MEMS microphones for biomedical applications

This paper introduces passive wireless telemetry based operation for high frequency acoustic sensors. The focus is on the development, fabrication, and evaluation of wireless, battery-less SAW-IDT MEMS microphones for biomedical applications. Due to the absence of batteries, the developed sensors are small and as a result of the batch manufacturing strategy are inexpensive which enables their utilization as disposable sensors. A pulse modulated surface acoustic wave interdigital transducer (SAW-IDT) based sensing strategy has been formulated. The sensing strategy relies on detecting the ac component of the acoustic pressure signal only and does not require calibration. The proposed sensing strategy has been successfully implemented on an in-house fabricated SAW-IDT sensor and a variable capacitor which mimics the impedance change of a capacitive microphone. Wireless telemetry distances of up to 5 centimeters have been achieved. A silicon MEMS microphone which will be used with the SAW-IDT device is being microfabricated and tested. The complete passive wireless sensor package will include the MEMS microphone wire-bonded on the SAW substrate and interrogated through an on-board antenna. This work on acoustic sensors breaks new ground by introducing high frequency (i.e., audio frequencies) sensor measurement utilizing SAW-IDT sensors. The developed sensors can be used for wireless monitoring of body sounds in a number of different applications, including monitoring breathing sounds in apnea patients, monitoring chest sounds after cardiac surgery, and for feedback sensing in compression (HFCC) vests used for respiratory ventilation. Another promising application is monitoring chest sounds in neonatal care units where the miniature sensors will minimize discomfort for the newborns.     

Development of porcine embryos reconstituted with somatic cells and enucleated metaphase I and II oocytes matured in a protein-free medium

Background

Many cloned animals have been created by transfer of differentiated cells at G0/G1 or M phase of the cell cycle into enucleated M II oocytes having high maturation/meiosis/mitosis-promoting factor activity. Because maturation/meiosis/mitosis-promoting factor activity during oocyte maturation is maximal at both M I and M II, M I oocytes may reprogram differentiated cell nuclei as well. The present study was conducted to examine the developmental ability in vitro of porcine embryos reconstructed by transferring somatic cells (ear fibroblasts) into enucleated M I or M II oocytes.

Results

Analysis of the cell cycle stages revealed that 91.2 ± 0.2% of confluent cells were at the G0/G1 phase and 54.1 ± 4.4% of nocodazole-treated cells were at the G2/M phase, respectively. At 6 h after activation, nuclear swelling was observed in 50.0-88.9% and 34.4-39.5% of embryos reconstituted with confluent cells and nocodazole-treated cells regardless of the recipient oocytes, respectively. The incidence of both a swollen nucleus and polar body was low (6.3-10.5%) for all nocodazole-treated donor cell regardless of the recipient oocyte. When embryos reconstituted with confluent cells and M I oocytes were cultured, 2 (1.5%) blastocysts were obtained and this was significantly (P < 0.05) lower than that (7.6%) of embryos produced by transferring confluent cells into M II oocytes. No reconstructed embryos developed to the blastocyst stage when nocodazole-treated cells were used as donors.

Conclusions

Porcine M I oocytes have a potential to develop into blastocysts after nuclear transfer of somatic cells.     

Increased oxidative stress in children with attention deficit hyperactivity disorder

Objectives: The purpose of this study was to investigate oxidative stress in children with attention deficit hyperactivity disorder (ADHD).

Methods: Total oxidant status (TOS), total antioxidant status (TAS), paraxonase-1 (PON-1) and arylesterase (ARE) activity were measured in 76 children (44 boys, 32 girls) diagnosed with ADHD according to the DSM-IV and 78 healthy children (46 boys, 32 girls).

Results: Age and sex were similar between the groups (P?>?0.05). TOS and the oxidative stress index (OSI) were higher in the patient group than the control group (P?<?0.001). PON-1 (P?=?0.002), ARE (P?=?0.010) activity and TAS (P?<?0.001) were lower in the patient group than the control group.

Discussion: We found decreased PON-1, ARE activity and TAS, and increased TOS and OSI in children with ADHD. Our study showed that there is significantly increased oxidative stress in children with ADHD.