Ovarian and Hormonal Responses to Follicular Phase Administration of Investigational Metastin/Kisspeptin Analog,TAK‐683, in Goats

This study evaluated the effects of follicular phase administration of TAK‐683, an investigational metastin/kisspeptin analog, on follicular growth, ovulation, luteal function and reproductive hormones in goats. After confirmation of ovulation by transrectal ultrasonography (Day 0), PGF_2α (2 mg/head of dinoprost) was administered intramuscularly on Day 10 to induce luteal regression. At 12 h after PGF_2α administration, intravenous administration of vehicle or 35 nmol (50 μg)/head of TAK‐683 was performed in control (n = 4) and treatment (n = 4) groups, respectively. Blood samples were collected at 6‐h intervals for 96 h and then daily until the detection of subsequent ovulation (second ovulation). After the second ovulation, ultrasound examinations and blood sampling were performed every other day or daily until the subsequent ovulation (third ovulation). Mean concentrations of LH and FSH in the treatment group were significantly higher 6 h after TAK‐683 treatment than those in the control group (12.0 ± 10.7 vs 1.0 ± 0.7 ng/ml for LH, 47.5 ± 28.2 vs 15.1 ± 3.4 ng/ml for FSH, p < 0.05), whereas mean concentrations of oestradiol in the treatment group decreased immediately after treatment (p < 0.05) as compared with the control group. Ovulation tended to be delayed (n = 2) or occurred early (n = 1) in the treatment group as compared with the control group. For the second ovulation, ovulatory follicles in the treatment group were significantly smaller in maximal diameter than in the control group (3.8 ± 0.5 vs 5.4 ± 0.2 mm, p < 0.05, n = 3). Administration of TAK‐683 in the follicular phase stimulates gonadotropin secretion and may have resulted in ovulation of premature follicles in goats.     

Extraction of lycopene from tomato skin with supercritical carbon dioxide: effect of operating conditions and solubility analysis

Supercritical carbon dioxide (SCCO2) extraction of lycopene from waste tomato skins was investigated. The experiments were carried out at pressures and temperatures ranging from 20 to 50 MPa and 313 to 373 K, respectively, without any modifiers. The flow rate of CO2 was maintained at 2.5 mL/min for 330 min extraction time. Solvent flow rate effect was examined for CO2 flow rates from 1.5 to 4.5 mL/min. The extracts were analyzed by high-performance liquid chromatography and UV-visible spectroscopy. The results showed that with optimized operating conditions, the maximum yield of lycopene (1.18 mg of lycopene/g of sample) was obtained at 40 MPa, 373 K, and 2.5 mL of CO2/min. Chromatographic analysis indicated that lycopene was extracted from tomato skin with negligible degradation at the optimum conditions and the amount extracted represented more than 94% of the total carotenoid content of the sample. The solubility of lycopene was modeled by use of the Chrastil equation.     

Distinct antigen recognition pattern during zoonotic visceral leishmaniasis in humans and dogs

Leishmania infantum is a causative agent of endemic zoonotic visceral leishmaniasis (VL) in regions of South America and the Mediterranean. Dogs are the major reservoirs for L. infantum in these regions, and control of disease in dogs could have a significant impact on human disease. Although dogs share many symptoms of VL with humans as a result of L. infantum infection, they also show some unique clinical manifestations, which are often a combination of visceral and cutaneous leishmaniasis, suggesting different mechanisms of disease development in dogs and humans. Here, we compare antibody responses of dogs and humans with VL to various defined leishmanial antigens. Parasite lysate and K39, the two most commonly used antigens for serodiagnosis of VL, detected the highest levels of antibodies in both humans and dogs with VL, whereas the recognition patterns of these antigens were distinct between the hosts. Among other defined antigens tested, LmSTI1 and CPB detected higher levels of antibodies in dogs and humans, respectively. These results indicate there is a difference between humans and dogs in antigen recognition patterns during VL. We infer that different strategies may need to be used in development of vaccines and diagnostics for humans and for dogs. In addition, we show a correlation between antibody titers to several antigens and severity of clinical symptoms during canine VL.     

Somaclonal variation as a source of resistance to eyespot disease of sugarcane

After 10 years of evaluation in different locations with high levels of incidence of disease, a group of sugarcane somaclones derived from callus tissues was selected for eyespot resistance. Resistance evaluations of four somaclones were performed under field and laboratory conditions. The results confirmed the superiority of two somaclones, one resistant and one tolerant to eyespot disease. Restriction analysis of mitochondrial DN A revealed that the two somaclones had a different DNA organization which distinguished them both from each other and from the donor plant; the restriction profile was similar however to that of the resistant control done. Restriction patterns of a third somaclone, also resistant, were similar to those of the donor plant. Differences among the somaclones were also evident when using a maize ribosomal DNA probe.     

Analyses of free amino acid and taste sensor traits in egg albumen and yolk revealed potential of value-added eggs in chickens

To create high-quality eggs by using different breed and feed materials, we investigated free amino acid contents and taste sensor traits using two chicken breeds (Rhode Island Red; RIR and Australorp; AUS) fed two feeds (mixed and fermented feeds). Two-way ANOVA revealed significant breed and feed main and interaction effects on albumen bitterness and a significant interaction effect on yolk bitterness. Albumen from RIR fed mixed feed and AUS fed fermented feed was higher bitterness, whereas yolk from those groups was lower bitterness. Significant breed effects were detected in four albumen amino acid traits (His, Met, Ile, and Lys) and a yolk His, whereas significant feed effects were found in 15 albumen amino acid traits (Asp, Glu, Ser, His, Gly, Thr, Ala, Tyr, Val, Met, Trp, Ile, Leu, Lys, and Pro) and a yolk cystine trait. Compared to albumen amino acids, yolk amino acids had limited effects by breed and feed. The present results suggest that genetic and nutritional factors can alter not only amino acid contents but also sensor values of bitterness, indicating that selecting the combination of breed and feed enable us to make amino acids enriched and taste added designer eggs in future.     

Growth prediction of young chicks: Do equal deficiencies of different essential amino acids produce equal growth responses?

1. Prediction of the growth of young chicks was attempted by means of a computerised mathematical model, by fitting a 4 parameter kinetic equation (Mercer, 1982) to data obtained experimentally and from the literature. Amino acid antagonisms between arginine and lysine, and between branched-chain amino acids were taken into account. 2. In order to pool the data, dietary concentrations of essential amino acids were standardised by expressing as percentages of requirements according to Japanese Feeding Standard (1984), and growth responses were expressed relative to growth at or above this requirement. 3. The kinetic model gave an excellent fit (R2 = 0.92-0.99) to independent growth data obtained experimentally and from the literature, validating the use of the equation. 4. It was implied from predicted growth that the responses to alterations in dietary concentrations of different essential amino acids differed substantially, in a characteristic pattern, even when percentages of requirements were the same. 5. The validity of the model was tested against data that were not used in its construction. It was found that the simulation model could predict the growth of young chicks satisfactorily from dietary amino acid patterns.     

Detection of equine immunoglobulin-secreting cells by a plaque assay.

A protein A-hemolytic plaque assay was applied to detect immunoglobulin (Ig)-producing cells in horse peripheral blood, using pokeweed mitogen as a B lymphocyte activator. A maximum number of Ig-secreting cells was obtained when horse peripheral blood lymphocytes were cultured in a medium containing horse serum. The number of Ig-secreting cells in young horses (2 years old) was lower than that in adult horses (6 to 23 years old). In addition, the plaque formation was unchanged from blood samples kept at 4 degrees C for 24 hours, while blood samples kept for 72 hours did not yield plaques. These results indicate that the plaque assay is a reliable and useful method for detecting Ig-secreting cells in the peripheral blood of the horse.     

Pathological studies on local tissue reactions in guinea pigs and rats caused by four different adjuvants.

We investigated pathological changes at the injection site in guinea pigs and rats for 16 weeks following a single intramuscular injection of one of the following oil adjuvant emulsions; oil adjuvant ISA-70, Freund's incomplete adjuvant, Freund's complete adjuvant, and aluminium phosphate gel. In the animals injected with ISA-70 emulsion prepared by manual shaking, grossly, there was partial thickening of subcutaneous tissue, discoloration of inter-muscular connective tissue, and swelling of the inguinal lymph nodes at 2 and 4 weeks post injection (PI). Histopathologically, ISA-70 injected sites revealed acute inflammatory changes at 72 hrs PI, and peak reactions consisting of macrophage accumulation around oil cysts and fibrosis were observed at 4 weeks PI. These changes were less severe and of shorter duration than those in the other three adjuvants. Guinea pigs and rats injected with materials containing inactivated Newcastle disease virus (NDV) antigen similarly showed an infiltration of plasma cells and lymphocytes in addition to the changes described above. ISA-70 containing NDV antigen induced similar hemagglutination-inhibition titer to that induced by Freund's incomplete adjuvant.     

Effects of cyclosporin A on susceptibility and interferon-generating capacity in mice infected with Toxoplasma gondii

Cyclosporin A (Cs-A), a potent immunosuppressant, was administered to mice to evaluate the role of T lymphocytes for the development of a protective immunity to an obligate intracellular protozoan parasite, Toxoplasma gondii. Although daily administration of various amounts of Cs-A for 7 days enhanced the host susceptibility at all doses employed, a dose-dependent manner of Cs-A treatment was not observed as far as the dosage regimens applied here are concerned; mice died at the same rate (40%) among the groups receiving various amounts of Cs-A. Cs-A treatment had a differential effect on the course of disease depending on how it was given in relation to infection. All mice receiving 50 mg of Cs-A per kg per day for 10 days from the beginning of infection eventually died of toxoplasmosis. Cs-A did not suppress the production of intereferon(IFN)-alpha/beta that was induced shortly after the infection, whereas it reduced greatly the ability of Toxoplasma-infected mice to produce IFN-gamma induced by stimulation with Toxoplasma lysate antigen (TLA). Moreover, the decrease in IFN-gamma production correlated with an increase in the parasitic growth in the peritoneal cavities of Cs-A-treated mice. These results suggest that the immunosuppressive effect of Cs-A on the primary Toxoplasma infection in mice is expressed by inhibiting the development of effector T cells responsible for the production of IFN-gamma.