Involvement of Ca2+-ATPase in suppressing the appearance of bovine helically motile spermatozoa with intense force prior to cryopreservation

In cattle, cryopreserved spermatozoa are generally used for artificial insemination (AI). Many of these specimens exhibit helical movement, although the molecular mechanisms underlying this phenomenon remain unclear. This study aimed to characterize helically motile spermatozoa, investigate the involvement of Ca²⁺-ATPase in suppressing the appearance of these spermatozoa prior to cryopreservation, and examine the potential of helical movement as an index of sperm quality. In the cryopreserved semen, approximately 50% of spermatozoa were helically motile, whereas approximately 25% were planarly motile. The helically motile samples swam significantly faster than those with planar movement, in both non-viscous medium and viscous medium containing polyvinylpyrrolidone. In contrast, in non-cryopreserved semen, planarly motile spermatozoa outnumbered those that were helically motile. Fluorescence microscopy with Fluo-3/AM and propidium iodide showed that flagellar [Ca²⁺]_i was significantly higher in cryopreserved live spermatozoa than in non-cryopreserved live ones. The percentage of non-cryopreserved helically motile spermatozoa was approximately 25% after washing, and this increased significantly to approximately 50% after treatment with an inhibitor of sarcoplasmic reticulum Ca²⁺-ATPases (SERCAs), “thapsigargin.” Immunostaining showed the presence of SERCAs in sperm necks. Additionally, the percentages of cryopreserved helically motile spermatozoa showed large inter-bull differences and a significantly positive correlation with post-AI conception rates, indicating that helical movement has the potential to serve as a predictor of the fertilizing ability of these spermatozoa. These results suggest that SERCAs in the neck suppress the cytoplasmic Ca²⁺-dependent appearance of helically motile spermatozoa with intense force in semen prior to cryopreservation.     

A case of canine multiple inflammatory colorectal polyps treated by endoscopic polypectomy and argon plasma coagulation

Endoscopic polypectomy and argon plasma coagulation (APC) were performed in a refractory case of inflammatory colorectal polyps in a 7-year-old male Miniature Dachshund. Colonoscopic examination revealed a large sessile polyp and multiple diffuse small polyps, localized to the descending colon and rectum. The case showed a poor therapeutic response to prednisolone and cyclosporine. Under anesthesia, piecemeal resections were performed by polypectomy. APC was carried out to cauterize the polyp remnants. After treatment, reduction of the lesions and the improvement in clinical signs were observed, without recurrence of lesions for at least 10 months. Endoscopic treatment by polypectomy and APC is suggested to be a therapeutic option for refractory cases of inflammatory colorectal polyps in dogs.     

A direct enzyme immunoassay for the measurement of furosemide in horse plasma

A new enzyme immunoassay (EIA) for the measurement of furosemide in horse plasma is described. The lower limit of detection of this EIA method was 7.8 ng/ml. The intra-and inter-assay coefficients of variation ranged from 2.5% to 4.9% and 7.5% to 9.8%, respectively. Cross-reactivity with other compounds was not observed. There was a high correlation (r2=0.987) between the high-performance liquid chromatography and EIA results obtained for furosemide concentrations in horse plasma. These results indicate that the newly developed EIA method is useful for the quantitative analysis of furosemide in horse plasma.     

Fipronil modulation of GABAA receptor single-channel currents

Fipronil is the first phenylpyrazole insecticide introduced for pest control. Although fipronil is known to inhibit GABA receptors, the detailed mechanism of action remains to be seen. In order to elucidate the mechanism of fipronil interaction with the mammalian GABAA system, single-channel patch clamp experiments were performed using rat dorsal root ganglion neurons. The amplitude of main conductance state (27pS) current was not significantly altered by co-application of 10 microM fipronil and 10 microM GABA. The histograms of open time distribution were fitted to a sum of three exponential functions. After application of 10 microM fipronil, the proportion of the fastest component increased slightly and that of the slowest component decreased slightly. Thus, the mean open time was decreased from 11.4 ms to 7.8 ms by fipronil. The histograms of closed time distribution were fitted to a sum of four exponential functions. Fipronil 10 microM prolonged the slowest time constant resulting in a prolongation of the mean closed time from 29.7 ms to 52.8 ms. Thus, the frequency of channel openings was reduced. Thus, the fipronil suppression of GABA-induced whole-cell currents is caused in part by decreases in the channel open time and the frequency of channel openings.     

Stress responses in neonatal meat and layer Nagoya chicks

We reported that meat chicks have either a greater capability to acclimatize to novel environments, or a blunted hypothalamic‐pituitary‐adrenal response to novel environments compared with layer chicks in a commercial base. The present study compared the differences in behavior and plasma corticosterone concentrations under isolation‐induced stress between neonatal meat and layer Nagoya chicks which had been separated from the same population. Both types of neonatal chicks reared in groups were individually separated and their spontaneous activity and distress‐induced vocalizations were monitored for 10 min. The responses of the two types were remarkably different, with the meat chicks being less active than the layer chicks. Distress‐induced vocalizations were fewer in the meat than in the layer chicks. The meat chicks spent more time in a sleeping posture during isolation‐induced stress. Plasma corticosterone concentrations measured at the end of the test tended to be higher in the layer chicks than in meat ones, but not significantly. In conclusion, the selection of Nagoya chickens for meat and layer may have trends similar to those observed in commercial chickens in relation to stress susceptibility.     

Refugee impact on collective management of forest resources: a case study of Bhutanese refugees in Nepal's Eastern Terai region

This article analyzes the relationship between forest resources, refugees, and the host population. The findings of the research suggest that the host population are heavily dependent on the local forest for their daily needs such as fuelwood, timber, grazing area, fodder for domestic animals, foods, and medicine in addition to cultural and esthetic needs. The forest has also been relied upon for agricultural needs such as manufacture of agricultural tools, maintenance of irrigation water systems, erosion control, and fertilizer needs. The forest was under a sustained demand as any other Terai forest of Nepal. After the arrival of refugees in 1992, the demand for forest resources increased substantially. Initially, the construction of the refugee camps decreased the total forest area and also required some felling of trees. More significantly, the refugees themselves became active users of the forest resource, which generated extra pressure on the forest and created scarcity of forest resources. Before the arrival of the refugees, forest management and monitoring of illegal use of the forest resources were carried out by the government through its local forester office. The local residents were active users of the forest resources, but were passive in managing and maintaining the forest resource. However, competition from the refugees instilled a desire in the local population to safeguard and protect the dwindling resource against the external threat by creating the Humse Dumse Community Forest.     

Spread of HTLV-I between lymphocytes by virus-induced polarization of the cytoskeleton

Cell contact is required for efficient transmission of human T cell leukemia virus- type 1 (HTLV-I) between cells and between individuals, because naturally infected lymphocytes produce virtually no cell-free infectious HTLV-I particles. However, the mechanism of cell-to-cell spread of HTLV-I is not understood. We show here that cell contact rapidly induces polarization of the cytoskeleton of the infected cell to the cell-cell junction. HTLV-I core (Gag protein) complexes and the HTLV-I genome accumulate at the cell-cell junction and are then transferred to the uninfected cell. Other lymphotropic viruses, such as HIV-1, may similarly subvert normal T cell physiology to allow efficient propagation between cells.     

Effects of Zeolite on Soil Nutrients and Growth of Barley Following Irrigation with Saline Water

ABSTRACT

Soil salinity is a major abiotic factor limiting crop production but an amendment with synthetic zeolite may mitigate effects of salinity stress on plants. The objective of the study was to determine the effects of zeolite on soil properties and growth of barley irrigated with diluted seawater. Barley was raised on a sand dune soil treated with calcium type zeolite at the rate of 1 and 5% and irrigated every alternate day with seawater diluted to electrical conductivity (EC) levels of 3 and 16 dS m^?1. Irrigation with 16 dS m^?1 saline water significantly suppressed plant height by 25%, leaf area by 44% and dry weight by 60%. However, a substantial increase in plant biomass of salt stressed barley was observed in zeolite-amended treatments. The application of zeolite also enhanced water and salt holding capacity of soil. Post-harvest soil analysis showed high concentrations of calcium (Ca^{2 +}), magnesium (Mg^{2 +}), sodium (Na⁺), and potassium (K⁺) due to saline water especially in the upper soil layer but concentrations were lower in soils treated with zeolite. Zeolite application at 5% increased Ca^{2 +} concentration in salt stressed plants; concentrations of trace elements were also increased by 19% for iron (Fe^{2 +}) and 10% for manganese (Mn^{2 +}). The overall results indicated that soil amendment with zeolite could effectively ameliorate salinity stress and improve nutrient balance in a sandy soil.