Effect of ovarian hormones on maturation of dendritic cells from peripheral blood monocytes in dogs

Previously, we reported that ovarian hormones affect the immune response against E. coli isolated from the dogs affected with pyometra. In order to investigate mechanisms underlying the immune modulation, we examined the effects of ovarian hormones on the generation of dendritic cells (DCs), the most potent antigen presenting cell. DCs were differentiated from peripheral blood monocytes (PBMOs) using a cytokine cocktail. Both estrogen receptor and progesterone receptors were expressed by the PBMOs and immature DCs. When various ovarian hormones were added to the culture for the DC differentiation, progesterone significantly decreased the expression of DC maturation markers, such as CD1a, CD80 and CD86, on mature DCs. Conversely, the addition of estrogen to the cultures increased the expression of CD86, but not other maturation makers. Furthermore, DCs differentiated in the presence of progesterone did not stimulate allogeneic mononuclear cells in PB. Taken together, these results indicate that progesterone diminishes the maturation of DCs, leading to decreased immune responses against invading pathogens.     

Mitigation of nitrous oxide (N2O) emission from swine wastewater treatment in an aerobic bioreactor packed with carbon fibers

Mitigation of nitrous oxide (N₂O) emission from swine wastewater treatment was demonstrated in an aerobic bioreactor packed with carbon fibers (CF reactor). The CF reactor had a demonstrated advantage in mitigating N₂O emission and avoiding NOx (NO₃ + NO₂) accumulation. The N₂O emission factor was 0.0003 g N₂O‐N/gTN‐load in the CF bioreactor compared to 0.03 gN₂O‐N/gTN‐load in an activated sludge reactor (AS reactor). N₂O and CH₄ emissions from the CF reactor were 42 g‐CO₂ eq/m³/day, while those from the AS reactor were 725 g‐CO₂ eq/m³/day. The dissolved inorganic nitrogen (DIN) in the CF reactor removed an average of 156 mg/L of the NH₄‐N, and accumulated an average of 14 mg/L of the NO₃‐N. In contrast, the DIN in the AS reactor removed an average 144 mg/L of the NH₄‐N and accumulated an average 183 mg/L of the NO₃‐N. NO₂‐N was almost undetectable in both reactors.     

Effect of lactoferrin on murine embryo development created from lipopolysaccharide-treated sperm

The effect of lactoferrin (LF) on embryo development was investigated by using lipopolysaccharide (LPS)-treated mouse sperm. For the development rate of the 2-cell stage embryo, the embryo derived from LPS- and LF-treated sperm showed similar survival rate to the control embryo. On day 12 after the embryo transfer into the recipient, the frequent abnormality was observed in the embryo derived from LPS-treated sperm, and the abnormality was tended to be inhibited in the embryo derived from LPS- and LF-treated sperm. These results imply that LF treatment on sperm contaminated with bacteria may facilitate the embryo development, which contribute to the improvement of infertility.     

Antioxidant constituents in distillation residue of Awamori spirits

Constituents in a distillation residue of Awamori (millet spirits) and their antioxidant activity are investigated in this study. The supernatant of the distillation residue obtained by centrifugation was partitioned with n-hexane, chloroform, ethyl acetate, and n-butanol against water to afford the corresponding solubles. Among them, n-hexane and chloroform solubles showed higher antioxidant potency than l-ascorbic acid by the bleomycin-Fe method. In chloroform solubles, seven cyclic dipeptides were identified along with ethyl 2-pyrrolidione-5-carboxylate, tyrosol, and ethyl p-hydoroxyphenyllactate. Antioxidant activity of ethyl p-hydoroxyphenyllactate was 4.2 times that of l-ascorbic acid, whereas cyclic dipeptides showed activity 0.89-1.29 times as strong as that of l-ascorbic acid. On the other hand, scavenging effect of cyclic dipeptides against O(2)(-.) and OH(.) by using electron spin resonance was also investigated. In the results, cyclo(l-Ile-l-Pro) showed significantly strong inhibitory effect against OH(.) (95.4% at 2.5 x 10-3 M) and cyclo(l-Phe-l-Pro), cyclo(l-Pro-l-Val), and cyclo(l-Leu-l-Pro) inhibited OH(.) 64.9, 54.1, and 51.0%, respectively, whereas alpha-tocopherol showed 37.7% inhibition, though only a few cyclic dipeptides weakly inhibited O(2)(-.).     

Specific detection of potentially allergenic kiwifruit in foods using polymerase chain reaction

Kiwifruit (Actinidia deliciosa and Actinidia chinensis) is allergenic to sensitive patients, and, under Japanese regulations, it is one of the food items that are recommended to be declared on food labeling as much as possible. To develop PCR-based methods for the detection of trace amounts of kiwifruit in foods, two primer pairs targeting the ITS-1 region of the Actinidia spp. were designed using PCR simulation software. On the basis of the known distribution of a major kiwifruit allergen (actinidin) within the Actinidia spp., as well as of reports on clinical and immunological cross-reactivities, one of the primer pairs was designed to detect all Actinidia spp. and the other to detect commercially grown Actinidia spp. (i.e., kiwifruit, Actinidia arguta, and their interspecific hybrids) except for Actinidia polygama. The specificity of the developed methods using the designed primer pairs was verified by performing PCR experiments on 8 Actinidia spp. and 26 other plants including fruits. The methods were considered to be specific enough to yield target-size products only from the target Actinidia spp. and to detect no target-size products from nontarget species. The methods were sensitive enough to detect 5-50 fg of Actinidia spp. DNA spiked in 50 ng of salmon testis DNA used as a carrier (1-10 ppm of kiwifruit DNA) and 1700 ppm (w/w) of fresh kiwifruit puree spiked in a commercial plain yogurt (corresponding to ca. 10 ppm of kiwifruit protein). These methods would be expected to be useful in the detection of hidden kiwifruit and its related species in processed foods.     

Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes

We analysed the effect of three antioxidants that have different functional mechanisms on the in vitro maturation (IVM) of porcine oocytes. Single oocyte monoculture using the hanging drop (HD) system has some advantages such as improving analysis efficiency brought by the smaller number of samples than the number of oocytes cultured in one drop. Direct effects of ligands on single oocytes could also be detected without considering the effects of paracrine factors from other oocytes. After 22 h of pre‐culture, denuded oocytes were cultured for 22 h with 0.01 and 0.1 μg/ml of L‐carnitine (LC), lactoferrin (LF) or sulforaphane (SF) in the presence/non‐presence of oxidant stress induced by H₂O₂ supplementation to evaluate the reducing effects against oxidative stress on nuclear maturation. As a result, compared with LC and SF, LF showed effective reduction in oxidative stress at a lower concentration (0.01 μg/ml), suggesting that LF is a more effective antioxidant in porcine oocyte IVM. Additionally, LF also increased maturation rate even in culture without H₂O₂. Our results clearly suggest that the HD monoculture system is useful for screening the substances that affect porcine oocyte culture.     

Utilization of free amino acids, yolk proteins and lipids in developing eggs and yolk-sac larvae of walleye pollock Theragra chalcogramma

ABSTRACT:   To elucidate the utilization of the major yolk nutrient stocks in eggs and larvae of walleye pollock Theragra chalcogramma , the contents of free amino acids (FAA), the major yolk protein (180 kDa lipovitellin originated from vitellogenin B in ovulated eggs: oLv B), and lipids were measured. Most eggs hatched 18 days after fertilization at 5°C, and all larvae absorbed almost all their yolk mass by 28 days. The total FAA content showed no change during the first 6 days, and then decreased to 28% of the initial level by 18 days. The oLv B contents, measured by an enzyme-linked immunosorbent assay using a specific antiserum against oLv B, gradually decreased from 6 to 18 days, followed by a rapid decline. The content of phospholipids (PL) and triacylglycerols (TG) showed no marked change until hatching, and then decreased until disappearance of yolk sac. From these results, it is proposed that there are two main periods for nutrient utilization in embryos and larvae of walleye pollock. In the first period, FAA was mainly utilized until 18 days after fertilization. Active utilization of oLv B and lipids (PL and TG) instead of FAA occurred during the second period from 18 to 28 days.     

Protein Requirements of the Freshwater Prawn Macrobrachium rosenbergii Evaluated by the Factorial Method

The dietary protein requirement of the freshwater prawn Macrobrachium rosenbergii to obtain the maximum body protein retention was assessed by the estimated protein needed daily for maintenance (M) and body protein increment (G). The body protein increment was estimated by quantifying body nitrogen increments when the prawn was fed high‐protein diets composed primarily of casein. The protein required for maintenance was estimated by regressing weight gains in feeding experiments using graded dietary levels of casein. The true daily retention (R) or increase in body protein of the prawn corresponds to the sum of M and G. The values of M, G, and R determined in the present study were 3.86, 0.24, and 4.10 g/kg body weight/d, respectively. On the basis of the daily body protein increment and the net protein utilization of casein (58%), dietary protein requirement of the juvenile M. rosenbergii for maximum body protein retention was suggested to be about 7.1 g/kg body weight/d.