Morphological analysis of viral communities in the floodwater of a Japanese paddy field

Capsid size distributions of viral communities in the floodwater of a Japanese paddy field under a long-term fertilizer trial were surveyed during the rice cultivation period by using transmission electron microscopy. The capsid size distributions were monophasic, and the median values fell within the range of 50-70 nm. The quartile intervals were narrow from either 40-50 nm or 50-60 nm to either 60-70 nm or 70-80 nm for most samples. There was no clear seasonal variation in the capsid size distribution of viral communities. The difference in size distributions among different fertilizer plots was also not found. Viral communities in the floodwater were predominated by isometric icosahedral forms. Viruses with octahedral capsids and elongated ones were rare and sporadic in the floodwater.     

Somatic cell and innate immune responses in mammary glands of lactating cows to intramammary infusion of Bifidobacterium breve at pre-drying off period

Intramammary infusion of Bifidobacterium breve (B. breve)-induced somatic cell (SC) counts, chemiluminescent response (CL), lactoferrin (LF) concentrations and mastitis-causing pathogens from quarters with subclinical mastitis were measured to evaluate innate immune response of mammary glands in dairy cows at 3 to 4 weeks before drying off. SC counts in 7 quarters of 7 control cows and 5 quarters of 6 cows with mastitis increased markedly on day 1 and SC values in control cows were significantly (P<0.05) increased and returned to pre-infusion levels on day 5 after B. breve-infusion. CL values in both groups increased markedly on day 1 and then decreased after B. breve-infusion; however, CL values in cows with mastitis did not return to normal levels on day 5 and at postpartum. The CL values were highly correlated with their SC counts in milk from both groups. LF concentrations increased toward day 3 after B. breve-infusion and were higher in cows with mastitis. B. breve-infusion eliminated 16.6% (1/6) of pathogens from 6 quarters with chronic subclinical mastitis. B. breve-induced SC responses in quarters from 3 cows with mastitis showed characteristic patterns of recovery, persistent and new infections. B. breve-induced SC counts in quarters from the cows in the pre-drying off were lower (25.7–70.6%) than those of the cows in mid-lactation. The intrinsic innate immune response in cows on pre-drying off may be decreased and appears to be insufficient to eliminate pathogens from mammary gland in the pre-drying off.     

Transport of subarctic large copepods from the Oyashio area to the mixed water region by the coastal Oyashio intrusion

The lateral transport of organic carbon in large grazing copepods ( Neocalanus cristatus, Neocalanus flemingeri, Neocalanus plumchrus and Eucalanus bungii ) from the Oyashio area to the mixed water region (MWR) by the coastal Oyashio intrusion was estimated using the data of VMPS (vertical multiple plankton sampler) and 1500 dbar-referred geostrophic transport from the CTD (conductivity temperature depth sensor) data of five cruises during June 2001 to April 2002 on a repeat observation section OICE (Oyashio Intensive observation line off Cape Erimo), which extends southeastward from Hokkaido Island, Japan. The transport to MWR by the coastal Oyashio intrusion was estimated to be 5.3 × 10¹¹ g C for the four species. Data from profiling floats also indicated that the copepods were advected from OICE to MWR by the coastal Oyashio intrusion within about 2 months. This transport is considered to be one of the significant sources of organic carbon in MWR as it is larger than the amount of large zooplankton consumed by Pacific saury ( Cololabis saira ) in MWR, one of the dominant copepod predators in this region.     

Aluminum toxicity of synthetic aluminum–humus complexes derived from non-allophanic and allophanic Andosols and its amelioration with allophanic materials

Natural non-allophanic Andosols often show aluminum (Al) toxicity to Al-sensitive plant roots. The significance of Al–humus complexes to Al toxicity has been emphasized. Allophanic Andosols also possess Al–humus complexes, but they rarely show any toxicity. In the present study, using model substances, we tested the toxicity of Al–humus complexes and its amelioration with allophanic materials. We extracted humic substances from the A horizons of a non-allophanic Andosol and an allophanic Andosol using a NaOH solution, and reacted the humic substances and partially neutralized AlCl₃ solution at pH 4. Allophanic material was purified from commercial Kanuma pumice. Plant growth tests were conducted using a medium containing the Al–humus complexes (50 g kg⁻¹), the allophanic material (0, 90, 180 and 360 g kg⁻¹) and perlite. The root growth of barley ( Hordeum vulgare L.) and burdock ( Arctium lappa ) was reduced in the media containing the Al–humus complexes derived from both the non-allophanic and allophanic Andosols when the allophanic material was not added. With the addition of the allophanic materials, particularly in the 360 g kg⁻¹ treatment, the growth of the barley roots was improved markedly. Although the root growth of the burdock tended to improve with allophanic materials, the effect was weaker than that for barley. Monomeric Al in a solution of the medium was not detected (< 0.05 mg L⁻¹) following the addition of 360 g kg⁻¹ of allophanic materials, whereas 0.8–1.7 mg L⁻¹ Al was recorded without the allophanic material.     

Deficits of learning and memory in Hemojuvelin knockout mice

Iron is involved in various physiological processes of the human body to maintain normalfunctions. Abnormal iron accumulation in brain has been reported as a pathogenesis ofseveral neurodegenerative disorders and cognitive impairments. Hemojuvelin (HVJ) is amembrane-bound and soluble protein in mammals that is responsible for the iron overloadcondition known as juvenile hemochromatosis. Although iron accumulation in brain has beenrelated to neurodegenerative diseases, it remains unknown the effect of mutation of HVJgene on cognitive performance. In our studies, HJV(−/−) mice showed deficits in novelobject recognition and Morris water maze tests. Furthermore, the expression ration ofapoptotic marker Bax and anti-apoptotic marker Bcl-2 in the hippocampus and prefrontalcortex showed higher levels in HJV(−/−) mice. Our results suggested that deletion of HJVgene could increase apoptosis in brain which might contribute to learning and memorydeficits in mutant mice. These results indicated that HJV(−/−) mice would be a usefulmodel to study cognitive impairment induced by iron overload in brain.     

Impaired female fertility in tubulointerstitial antigen-like 1-deficient mice

Tubulointerstitial nephritis antigen-like 1 (Tinagl1, also known as adrenocortical zonation factor 1 [AZ-1] or lipocalin 7) is a matricellular protein. Previously, we demonstrated that Tinagl1 expression was restricted to extraembryonic regions during the postimplantation period and detected marked expression in mouse Reichert’s membranes. In uteri, Tinagl1 is markedly expressed in the decidual endometrium during the postimplantation period, suggesting that it plays a physical and physiological role in embryo development and/or decidualization of the uterine endometrium during pregnancy. In the present study, in order to determine the role of Tinagl1 during embryonic development and pregnancy, we generated Tinagl1-deficient mice. Although Tinagl1^–/– embryos were not lethal during development to term, homologous matings of Tinagl1^–/– females and Tinagl1^–/– males showed impaired fertility during pregnancy, including failure to carry pregnancy to term and perinatal lethality. To examine ovarian function, ovulation was induced with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG); the number of ovulated oocytes did not differ between Tinagl1^–/– and Tinagl1^flox/flox. In vitro fertilization followed by embryo culture also demonstrated the normal developmental potential of Tinagl1-null embryos during the preimplantation period. Our results demonstrate that Tinagl1 deficiency affects female mice and results in subfertility phenotypes, and they suggest that although the potential of Tinagl1^–/– oocytes is normal, Tinagl1 is related to fertility in adult females but is not essential for either fertilization or preimplantation development in vitro.     

Glomerular lipidosis accompanied by renal tubular oxalosis in wild and laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)

Glomerular lipidosis is a disease characterized by lipid accumulation in mesangial cells but that has not been fully investigated in avian species. We examined four wild and two laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)--an endangered avian species--presenting vacuolar deposits in the glomeruli. All cases had vacuolar deposits in the glomeruli. In the wild cases, fewer than 30% of all glomeruli were affected, compared with more than 90% in the laboratory-reared cases. In the wild cases, most deposits were mild and restricted to the mesangial areas of glomeruli. In the laboratory-reared cases, nearly all of the deposits covered entire glomeruli. Electron microscopy of mild deposits revealed vacuoles in the cytoplasm of mesangial cells. These vacuoles were positive for Sudan III, Sudan black B, oil red O, Nile blue, periodic acid-Schiff, Schultz test, and digitonin stain and were negative for performaric acid-Schiff stains. Based on these results, we diagnosed the glomerular lesion as glomerular lipidosis caused by uptake of low-density lipoprotein in mesangial cells. Except for one wild case, all cases exhibited renal tubular oxalosis. The severity of tubular oxalosis tended to be related to the severity of glomerular lipidosis: In cases of mild glomerular lipidosis, tubular oxalosis was also mild or absent. We therefore diagnosed the primary lesion as glomerular lipidosis accompanied by tubular oxalosis. The four wild cases came from different zones and therefore had no opportunities to interbreed and no common relatives. We believe these data support the hypothesis that glomerular lipidosis is a disease of the general population ofJapanese rock ptarmigans. This is the first report of glomerular lipidosis accompanied by renal tubular oxalosis in an avian species.     

Nuclear transfer using a bovine mammary epithelial cell line (BMEC)

The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10⁵ cells/cm² (high‐density group) or 0.8 × 10⁴ cells/cm² (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly (P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.