Identification of anthocyanins in the sprouts of buckwheat

The anthocyanin profiles and varieties/breeding line differences of anthocyanin concentrations in common/tartary buckwheat sprouts have been studied. Four anthocyanins, cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-galactoside, and cyanidin 3-O-galactopyranosyl-rhamnoside, were isolated from the sprouts of common buckwheat, were separated using high-performance liquid chromatography (HPLC), and were identified using reversed-phase liquid chromatography (LC)/electrospray ionization-mass spectrometry (ESI-MS)/MS techniques. In tartary buckwheat sprouts, two anthocyanins (cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside) were identified. Among 19 common/tartary buckwheat varieties/breeding lines, Hokkai T10 contained the highest amounts of anthocyanins. Cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside concentrations in 6-10 days after seeding sprouts of Hokkai T10 ranged from 0.16 to 0.20 mg/g dry wt and from 5.55 to 6.57 mg/g dry wt, respectively. In addition, dark-grown sprouts of Hokkai T10 accumulated 0.091 and 2.77 mg/g dry wt of cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside whereas other varieties/breeding lines accumulated trace amounts of anthocyanins. Given its anthocyanin-rich red cotyledons, Hokkai T10 is a promising line for use as "Moyashi" type sprouts and is strongly recommended as a new functional food, rich in dietary anthocyanins.     

Spontaneous Adenosquamous Carcinoma with Rapid Growth and EMT-like Changes in the Mammary Gland of a Young Adult Female BALB/c Mouse

This study histopathologically and immunohistochemically investigated a spontaneously occurring single mass subcutaneously located in the left lower abdomen of a female BALB/cAJcl−nu/+ mouse at 10 weeks of age. The mass was about 20 × 15 × 10 mm in size after formalin fixation; nevertheless, it was not detected by clinical observations at 9 weeks of age. H&E staining revealed the tumor origin was epithelial and probably arose from the mammary gland, and the tumor cells demonstrated a squamous, acinar or polyhedral/basal pattern. A cell kinetics analysis revealed that many of the tumor cells of the squamous, acinar or polyhedral/basal component were positive for PCNA and cyclin D1, although there were a few of TUNEL-positive tumor cells in all of the components. An epithelial/mesenchymal analysis demonstrated that most of the tumor cells of the squamous and acinar components contained keratin and E-cadherin; however, most of the tumor cells of the polyhedral/basal component were less or very weakly positive for these markers. The tumor cells of the squamous component were negative for vimentin and SMA; however, many of the tumor cells of the polyhedral/basal component exhibited vimentin. In addition, expression of SMA was confirmed in some tumor cells of the acinar and basal components. Based on the microscopic and immunohistochemical characterizations, the tumor was diagnosed to be adenosquamous carcinoma that originated from the mammary gland with rapid growth, and the tumor cells demonstrated epithelial-mesenchymal transition-like changes.     

Birth of piglets through the non-surgical transfer of blastocysts produced in vitro

In this study, we attempted to produce piglets by non-surgically transferring blastocysts produced in vitro, using a flexible catheter as the transfer instrument. Cumulus-oocyte complexes (COCs) were aspirated from the follicles of ovaries obtained at a local slaughterhouse. They were then matured in modified North Carolina State University (NCSU)-37 medium for 44-46 h and fertilized in porcine gamete medium (PGM). Ten hours after in vitro fertilization (IVF), presumptive zygotes were removed from the cumulus cells and cultured in porcine zygote medium (PZM)-5. Blastocysts were cultured for five days after IVF and, using a catheter for deep intrauterine insemination without sedation, they were transcervically transferred into the uterine horn of six recipients (45-50 blastocysts/recipients) whose estrous cycles were synchronized, at 5 days after human chorionic gonadotropin (hCG) injection. Of the six recipients, one sow became pregnant and farrowed seven piglets (four live piglets) 119 days after hCG injection. The body weight at birth of the newborns ranged from 0.8 to 1.4 kg. These results indicate that it is possible to obtain piglets by transcervically transferring blastocysts produced by IVF and in vitro cultures in chemically defined media.     

A time-course study of flavonoids in the sprouts of tartary (Fagopyrum tataricum Gaertn.) buckwheats

The evolution, from 1 to 10 days after germination, of flavonoid content in sprouts of tartary buckwheat (Fagopyrum tataricum Gaertn.), grown in a greenhouse under low light conditions (16 μmol m⁻² s⁻¹), was investigated. Chlorogenic acid and flavonoids including C-glycosylflavones (orientin, isoorientin, vitexin, isovitexin), rutin and quercetin were separated from the sprouts by HPLC and quantified with their commercial standards. Rutin content in the edible portion of the sprouts (mean 20 and 37 mg g⁻¹ DW in ‘Hokkai T 8’ and ‘Hokkai T 10’, respectively) was 3- to 31-fold greater than that in the roots or pericarp. The free radical scavenging activity of seed sprouts was assessed through the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. From 6 to 10 days after sowing, the free radical scavenging activity in the edible portions rose significantly from 1.52 to 2.33 μmol Trolox equiv. g⁻¹ DW in ‘Hokkai T 8’ and from 1.46 to 2.09 μmol Trolox equiv. g⁻¹ DW in ‘Hokkai T 10’, but differences between ‘Hokkai T 8’ and ‘Hokkai T 10’ were not significant. As the results, the sprouts of tartary buckwheat, particularly those of ‘Hokkai T 10’ are strongly recommended as new high rutin food.     

Effects of leptin and leptin peptide amide on the release of luteinizing hormone, growth hormone and prolactin from cultured porcine anterior pituitary cells

The present study was carried out to determine whether leptin or leptin (116–130) peptide amide (lep (116–130)), an active fragment of the native protein in rats, is able to stimulate the release of luteinizing hormone (LH), growth hormone (GH) or prolactin (PRL) from cultured porcine anterior pituitary (AP) cells in vitro. The AP cells were obtained from 6 month‐old pigs and were incubated for 3 h with 10^?11?10^?7 mol/L leptin or lep (116–130) after being cultured in Dulbecco's modified Eagle's medium for 3–4 days. Leptin significantly increased the concentration of LH and GH in the culture medium at concentrations of 10^?8 and 10^?7 mol/L, respectively, compared with the controls (P < 0.05). Leptin did not increase the concentration of PRL in the culture medium. In contrast to these results, no effects of lep (116–130) on the release of LH, GH or PRL were seen in the cultured cells. These results suggest that leptin stimulates the release of LH and GH by acting directly on porcine AP cells, and that a fragment of leptin protein comprising amino acids 116–130 is not associated with the secretion of hormones in pigs.     

Effects of intracerebroventricular injections of leptin on the release of luteinizing hormone and growth hormone in castrated calves

The purpose of the present study was to clarify the hypothalamic action of leptin on the secretion of luteinizing hormone (LH) and growth hormone (GH) in cattle. Intracerebroventricular (the third ventricle) injections of leptin were given to fully fed castrated Holstein calves. Blood samples were collected at 10‐min intervals for 60 min after injection and 20‐min intervals for 60 min before injection and for 60–180 min after injection through an indwelling catheter in the external jugular vein. Plasma LH and GH levels were examined by homologous radioimmunoassay. The administration of 10 µg of leptin stimulated a significant (P < 0.05) release of GH but not LH. Average GH levels began to rise after 30 min and were significantly increased at 40, 50 and 60 min after the injection, compared with the respective control values (P < 0.05). The present result suggests that leptin may act partly on the hypothalamus to stimulate the release of GH in castrated calves.