Implications of ram sperm rheotaxis analysed by microfluidics for fertility

Rheotaxis of sperm using a microfluidic device was explored in human, mice and bull. However, the rheotaxis of ram sperm and its role in fertility are unknown. Herein, we described the sperm rheotaxis in ram using microfluidic devices and focused on rheotaxis as potential markers of in vivo fertility. Computer-assisted sperm analysis (CASA) with controlled flow velocity was used to explore the kinematic parameters of sperm, total motility and positive rheotaxis (PR). The percentage of PR was defined as the number of PR sperm cells over the number of motile sperm cells. Then, according to the percentage of PR sperm, rams were classified into two groups; sperm with ≥40% PR and <40% PR, although the two ram groups showed similar total motility and kinematic values of sperm evaluated by CASA (p > .05). Two groups of rams mated one hundred thirty ewes naturally (10 ewes/ram). In the results, the pregnancy rate was higher in ≥40% PR (94.4%) than in <40% PR (42.5%, p < .05) after natural mating. Besides, the pregnancy loss was higher in <40% PR (33.3%) than in >40% PR group (8.1%, p < .05). In conclusion, the PR examination in semen can contribute to evaluate the reproductive performance of ram that will provide valuable insights into the semen evaluation.     

Matrix metalloprotease-9 activity in the cerebrospinal fluid and spinal injury severity in dogs with intervertebral disc herniation

We investigated whether matrix metalloproteinase (MMP)-9 expression in the cerebrospinal fluid (CSF) of dogs with intervertebral disc herniation (IVDH) is associated with the severity of neurological signs and prognosis. CSF from the cisterna magna (C-CSF) and the lumbar spine (L-CSF) of 34 dogs with IVDH was analyzed using zymography. Activity of MMP-9 in L-CSF was detected in 6 of 34 dogs with IVDH, often for more than 7 days after injury. MMP-9 activity was not detected from any of the C-CSF samples. Of the six cases that were MMP-9 positive, all four cases with grade V that had loss of deep pain were non-ambulatory 6 months after treatment. The remaining two cases with grade III and IV could recover mobility. In dogs with grade V thoracolumbar IVDH, MMP-9 expression in the CSF may indicate severe spinal cord injury with poor prognosis.     

Evaluation of the Affinity of Some Toxic Elements to Schwertmannite in Natural Streams Contaminated with Acid Mine Drainage

In order to evaluate fixation potential of schwertmannite for fluvial transport of various toxic elements, we examined bottom precipitates and stream waters collected from the rivers contaminated with acid mine drainage (AMD), which arose from the abandoned Nishinomaki mine (Shimonita, Gunma, Japan). Mineralogical and morphological observations revealed that schwertmannite was the main mineral of the precipitates. The affinity of various toxic ions to schwertmannite was evaluated on the basis of (1) apparent solid?Cliquid partition coefficients (K _d??s) between precipitates and stream waters, (2) coprecipitation behaviors during schwertmannite formation in a laboratory test, and (3) consideration on coprecipitation processes using partial charge model (PCM). As a result, oxyanions of V, As, Mo and Sb, K _d??s of which were relatively large (>10⁴ (ml g^?1)), were considered to be immobilized by schwertmannite precipitates. A laboratory test also demonstrated that these ions except Mo coprecipitated with schwertmannite. In addition, partial charges and average electronegativities predicted on the basis of PCM suggested that the oxyanions of V, As, Mo, and Sb could create stable inner sphere complexes with schwertmannite embryos, which results in their high affinity to schwertmannite. On the other hand, cationic ions of Mn, Cu, Zn, Sr, Cs, and U, K _d??s of which were relatively small (<10⁴ (ml g^?1)), were thought to have a tendency to flow downstream without uptake by schwertmannite precipitates. All these results suggested that schwertmannite has high fixation potential for fluvial transport of various toxic oxyanions in AMD-contaminated rivers.     

Contents of heavy metal elements in copper mosses: Scopelophila ligulata,Scopelophila cataractae,and Mielichhoferia japonica and their substrates

Scopelophila ligulata (Spruce) Spruce, S. cataractae (Mitt.) Broth., and Mielichhoferia japonica Besch. (Mielichhoferia mielichhoferi (Hook.) Wijk et Marg. var. japonica (Besch.) Wijk et Marg.) are well known “copper mossess” which show peculiar ecological characteristics consisting of a preference for metal-rich soils, lithosols, rocks, etc. as their substrates. The total and water-soluble element contents in the substrates and heavy metal contents in the moss body were analyzed in this study. The results of these chemical analyses were as follows: The iron contents in the moss body and substrate of S. ligulata were higher than those of S. cataractae and M. japonica. On the contrary the copper contents of the former were lower than those of the latter. The pH values of substrate of S. ligulata were lower than those of S. cataractae and M. japonica. The concentrations of water-soluble iron and sulfate in the substrate of S. ligulata were higher than those of S. cataractae and M. japonica. On the contrary, the water-soluble copper concentrations in the substrate of S. ligulata were lower than those of the others. Although in the substrate of S. cataractae the iron content was several times higher than the copper content, in the moss body the copper content was higher than iron content.     

Carbon and nitrogen contents and greenhouse gas fluxes of the Eurasian steppe soils with different land-use histories located in the Arkaim museum reserve of South Ural,Russia

The effects of different land-use histories on contents of soil carbon (C) and nitrogen (N) and fluxes of greenhouse gases [carbon dioxide (CO₂), methane (CH₄), and nitrous oxide (N₂O)] measured using the closed chamber method were investigated in the Arkaim museum reserve located in the South Ural of Russia. A natural forest site (NF) and two grassland sites that had different land-use histories (CL: cropland until 1991; PST: pasture until 1991; both sites have been fallow for 18 years) were selected for soil sampling and gas flux measurements. The vegetation in NF was mainly Betula pendula Roth. with steppe cherry and grassy cover. Perennial grasses (Stipa spp., Festuca spp. and others) have been planted in CL and PST since 1991 to establish reserve mode, and the projective cover of these plants were?>?90% in both sites in 2009. Soil samples were taken from the A horizon in the three sites, and additionally samples of the O horizon were taken from NF. The contents of soil C and N [total C, total N, soluble organic C, soluble N and microbial biomass C (MBC)] in the O horizon of NF were the largest among all investigated soils (p?p?2 fluxes (i.e., CO₂ efflux) in all three investigated sites were observed. The CO₂ efflux in NF was significantly larger than in CL and PST (129, 30 and 25?mg C m^?2 hour^?1, respectively, p?2 efflux between CL and PST. There were no significant differences in the fluxes of CH₄ and N₂O among NF, CL and PST (p?>?0.05). Our current research indicated that, in soils of the Eurasian steppe zone of Russia, total C, total N and MBC were affected not only by current land-use (i.e., fallow grassland vs. natural forest) but also by past (until 18 years ago) land-use.     

Effects of rearing water quality on expression of brain corticotropin-releasing hormone and gonadal development in goldfish (Carassius auratus)

The effects of changes in water quality in a recirculating system on brain corticotropin-releasing hormone (CRH) gene expression in goldfish were examined. In the experimental group, fish were reared without water change, while in the control group, 50% of water was changed everyday. In the experimental group, total ammonia-nitrogen, nitrate and phosphate were increased over the experimental period. Brain CRH mRNA levels in the control group seemed higher than those in the experimental group, but differences were confounded by variation among individuals. There were no differences in gonadal development between both groups.     

Early dispersal of ayu during marine stages as inferred from geographic variation in the number of vertebrae

ABSTRACT:   Marine environments often allow interbreeding of individuals over the species range, and analyses using neutral molecular markers may lose extant genetic boundaries laid between geographic majorities. Ayu Plecoglossus altivelis has a typical amphidromous life history, migrating between rivers and the sea. In order to clarify reproductive elements of the species, migrants from 64 rivers and streams sampled over a wide latitudinal range were examined for their vertebral number (VN) and dorsal pterygiophore number (DPN) as morphological markers to estimate the temperature history. The irregular variability in VN without a geographic cline suggests that the ascending schools of fish are composed of conspecifics sharing incubation temperature or hatching site. The intersample difference in DPN indicates that larvae and juveniles of ayu spend their marine life inside a water body with a distinct temperature. Site tenacity during the marine stage may be helpful to enhance the opportunity to return to the river where they hatched. Each assemblage of fish ascending to rivers and streams is deemed to mostly represent a reproductive element, and therefore, each acts as an evolutionarily significant unit within a metapopulational structure.     

Structural basis of biological N2O generation by bacterial nitric oxide reductase

Nitric oxide reductase (NOR) is an iron-containing enzyme that catalyzes the reduction of nitric oxide (NO) to generate a major greenhouse gas, nitrous oxide (N(2)O). Here, we report the crystal structure of NOR from Pseudomonas aeruginosa at 2.7 angstrom resolution. The structure reveals details of the catalytic binuclear center. The non-heme iron (Fe(B)) is coordinated by three His and one Glu ligands, but a His-Tyr covalent linkage common in cytochrome oxidases (COX) is absent. This structural characteristic is crucial for NOR reaction. Although the overall structure of NOR is closely related to COX, neither the D- nor K-proton pathway, which connect the COX active center to the intracellular space, was observed. Protons required for the NOR reaction are probably provided from the extracellular side.