A potential live vector, foamy virus, directed intra-cellular expression of ovine interferon-tau exhibited the resistance to HIV infection

Interferon-tau (IFN-tau), produced by the embryonic trophectoderm, is a member of type I IFNs required for the establishment of pregnancy in the ruminant ungulates. Although this IFN possesses antiviral activity similar to other type I IFNs, the effectiveness of IFN-tau as an antiviral agent has not been well characterized. To investigate possible antiviral effects of ovine IFN-tau (oIFN-tau), oIFN-tau-GST fusion protein was expressed in E. coli BL21, from which the purified protein isolated possessed anti-viral activity. An apathogenic human foamy virus (hFV) was then used to establish a potential recombinant live vector consisting of oIFN-tau cDNA sense (+) or antisense (-) sequence, oIFN-tau(+)/hFV or oIFN-tau(-)/hFV, respectively. Human hematopoietic and other mammalian cell lines that had been transduced with hFV vector consisting of no oIFN-tau, oIFN-tau(+)/hFV or oIFN-tau(-)/hFV construct were cultured initially for 12 days, and three of cell lines were then maintained for up to 90 days. These cells with oIFN-tau expression directed by hFV exhibited the in vitro cytopathic effect minimally. Transduced cell lines that had been cultured for 90 days were subjected to studies on human immunodeficiency virus type-1 (HIV-1) infection, which was measured with infectivity of viral particles resulted from the GFP inserted T-cell tropic HIV SF2 or macrophage tropic HIV SF162: the number of HIV-1 positive cells was reduced by the hFV driven-intra-cellular oIFN-tau expression. Since oIFN-tau/hFV transduced cells exhibited the resistance to HIV-1 infection and/or replication, oIFN-tau could be considered as one of effective antiviral agents against HIV-1. These results suggest that the hFV genome could be an effective recombinant live vector for the expression of a targeted gene in various cell types.     

T-cell lymphoma in a wild Okinawa rail (Gallirallus okinawae)

The Okinawa rail (Gallirallus okinawae) is an endangered species that inhabits the northern part of Okinawa Main Island in southern Japan. A wild Okinawa rail was rescued from a road in Kunigami Village in Okinawa in October 2009. The bird subsequently died and underwent necropsy. Tumors were found in the liver, spleen and part of the small intestine. Microscopically, lymphoid neoplasm was confirmed in these tissues. The tumor cells were mainly positive for CD3 and CD8α by immunohistochemistry. No Marek's disease virus genes were detected by PCR of a liver tumor. This is the first report of T-cell lymphoma in the Okinawa rail.     

Growth and metabolic characterization of Macrorhabdus ornithogaster.

Macrorhabdus ornithogaster (M. ornithogaster) is an anamorphic ascomycetous yeast found only in the stomach of birds. Infection is often benign but has also been associated with disease in some species of birds under some circumstances. In vitro efforts to grow M. ornithogaster have been largely unsuccessful. In this report, multiple liquid and solid media of varying pH, sugar concentration, and fetal bovine serum (FBS) concentrations, incubated at various temperatures in room air or microaerophilic conditions, were examined for their ability to support the growth of M. ornithogaster, obtained from a budgerigar (Melopsittacus undulatus). Optimum growth conditions were found to be Basal Medium Eagle's, pH 3 to 4, containing 20% FBS, and 5% glucose or sucrose under microaerophilic conditions at 42 degrees C. Using these conditions, M. ornithogaster was repeatedly passaged without loss of viability. Polyclonal isolates of M. ornithogaster consistently assimilated glucose, sucrose, and trehalose. M. ornithogaster did not grow with prolonged exposure to atmospheric oxygen, but growth in microaerophilic conditions was moderately enhanced by preincubation with atmospheric oxygen for 24 hours. An isolate of M. ornithogaster was found to be infective to day-old chickens, reduce their rate of weight gain, and induce a mild to moderate heterophilic inflammation of the isthmus. M. ornithogaster was reisolated from the chicks 7 days after infection, fulfilling Koch's postulates. A 761-bp sequence of 18S rDNA from this isolate was compared to the originally reported M. ornithogaster sequence and was found to be 97% identical.     

Identification and morphological characteristics of dental neonatal line in sika deer (Cervus nippon)

The dental neonatal line of the sika deer (Cervus nippon) was identified experimentally using chronological labeling methods. In the enamel, prominent dark lines were observed under transmitted light, and the number of increments between the dark line and labeling line was almost consistent with the day-age at the time of labeling injection. Therefore, we identified the dark line as the enamel neonatal line. In the dentin, the bright line was observed under polarized light. Since the bright line corresponded to the enamel neonatal line, we recognized the bright line as the dentin neonatal line. Neonatal lines intersected with the enamel-dentin junction at approximately one-third cervical in the first molar. Using these features, it would make possible to distinguish the neonatal line in wild sika deer.     

Insulin and glucagon secretory patterns during propionate and arginine tolerance tests in Japanese black cattle with growth retardation

To evaluate the energy condition of cattle with growth retardation, propionate (PTT) and arginine tolerance tests (ATT) were carried out. The insulin/glucagon concentration ratio immediately before PTT or ATT in the cattle with growth retardation was lower than in the control. In the growth-retarded cattle, insulin-AUC(0-120 min) during PTT was lower than in the control, while glucagon-AUC(0-120 min) was the same as in the control. Insulin-AUC(0-120 min) during ATT in the cattle with growth retardation tended to be lower than in the control, whereas glucagon-AUC(0-120 min) was the same. Therefore, insulin-AUC(0-120 min)/glucagon-AUC(0-120 min) in the cattle with growth retardation was lower than in the control during both tolerance tests. The growth-retarded cattle showed lower insulin/glucagon ratio similar to that found in starved and lactating cattle, suggesting a lack of energy.     

Development of a CO2 gas analyzer for monitoring soil CO2 concentrations

Measurement of soil CO₂ concentrations is important for investigating the dynamics and diffusion of CO₂ in soil. In this study, we developed a small CO₂ analyzer for measuring in situ-soil CO₂ concentrations. The CO₂ analyzer consists of a module containing an infrared CO₂ gas sensor, a temperature sensor, and a relative humidity sensor. These sensors are installed in a protective box with an air vent, which is suitable for burying in the soil. The output response time of the CO₂ analyzer was 349 s, as evaluated from the phase lag after input of known CO₂ concentrations. This response time is short enough to measure soil CO₂ concentrations, because variations in concentration are slower than the response time of the analyzer. In a field test, we used the CO₂ analyzer to measure soil CO₂ concentrations at five depths (0–50 cm) over 2.5 months. While the CO₂ concentration generally increased with depth, the amplitude of the variation in CO₂ concentration decreased with depth. The phase lag of the variations in soil CO₂ concentration also increased with depth, as did soil temperature. The tests confirm that the CO₂ analyzer is applicable to continuous monitoring of soil CO₂ concentrations.     

Carbon balance in a cool–temperate deciduous forest in northern Japan: seasonal and interannual variations, and environmental controls of its annual balance

We monitored variation in seasonal and annual net ecosystem production (NEP), gross primary production (GPP), and ecosystem respiration (R _E) based on 7-year eddy covariance measurements above a cool?Ctemperate deciduous broad-leaved forest (Japanese beech forest). The 7-year means (±SD) of annual NEP, GPP, and R _E were 312?±?64, 1250?±?62, and 938?±?36?g?C?m^?2?year^?1, respectively. Variation in NEP was much larger than variation in GPP and R _E. During the growing season, the main factor controlling carbon balance was air temperature; variation in seasonal integrated NEP was regulated by accumulated air temperature (degree-day) with a significant negative correlation, whereas the seasonal ratio of R _E to GPP was correlated positively with accumulated air temperature. Because the deviation of seasonal NEP was also significantly correlated with seasonal R _E/GPP, NEP was controlled by R _E/GPP, depending on air temperature during the growing season. Seasonal R _E in the defoliation and snow seasons was also important for evaluating the annual carbon balance, because the total number of days in the two seasons was quite large owing to a long snowy winter. In the defoliation and snow seasons, we found defoliation season length was a major factor determining seasonal integrated R _E, illustrating the positive correlation between R _E and defoliation season length. The major factors controlling interannual variations in forest carbon balance are discussed.     

Composition of culture medium influences zoosporogenesis and differentiation of <Emphasis Type="Italic">Aphanomyces cochlioides</Emphasis>

A modified medium was used to culture mycelium and produce a large quantity of zoospores of Aphanomyces cochlioides, a principal pathogen of damping-off disease of sugar beet and root rot disease of spinach. The semisolid medium consisted of 17 g corn meal agar (CMA) added with 4 g of yeast extract (YE) per liter of 50 mM phosphate buffer (pH 6.8–7.0). This medium supported the production of ca. 10⁶ zoospores ml⁻¹ in 6-day-old cultures, approximately 11-fold higher than the commonly used CMA (17 g CMA per liter of water, pH 6.0 ± 0.2). Although morphological characters of the zoospores produced from the hyphae grown on CMA and CMA + YE were almost similar, they contrasted their developmental strategy after encystment induced by mechanical agitation. Cystospores originating from the zoospores on CMA regenerated zoospores (>80%), while those from CMA + YE germinated (ca. 80%) and produced hyphae. Furthermore, 4–10% of the germinated cystospores on CMA + YE had double germ tubes. The soluble protein profiles of zoospores produced on CMA and on CMA + YE demonstrated that several proteins were either different or expressed differently. Our results suggest that the culture medium directly influences zoosporogenesis in A. cochlioides hyphae and the developmental strategy of the produced zoospores.