Rapid determination of the main organic acid composition of raw Japanese apricot fruit juices using near-infrared spectroscopy

The potential of near-infrared (NIR) spectroscopy to rapidly determine citric and malic acid contents of raw Japanese apricot (Japanese "ume", also known as the Japanese plum) fruit juice was investigated. In total, 314 raw juice samples with different organic acid compositions were collected over a long growth period, and spectra (1100-1850 nm) of these samples were acquired using an NIR spectrophotometer with a 1-mm path length. Calibrations were performed using a partial least-squares regression method based on a calibration sample set (211 samples), while validations were performed based on a validation sample set (103 samples). The results revealed good agreement between NIR spectroscopy and capillary electrophoresis, including the correlation coefficient (r2), standard error of prediction (SEP), and bias; no statistically (p = 0.05) significant differences were found for these parameters. Moreover, standard deviation ratios of reference data in the validation sample set to the SEP were higher than 3, indicating that NIR spectroscopy may represent an acceptable method for quantitative evaluation of citric and malic acids in raw Japanese apricot fruit juice.     

Possible association of glaucoma with pectinate ligament dysplasia and narrowing of the iridocorneal angle in Shiba Inu dogs in Japan

The purpose of the study was to describe the shape of the pectinate ligament (PL) and to estimate the width of the iridocorneal angle (ICA) in glaucomatous and non-glaucomatous eye of Shiba Inu dogs in Japan. One hundred fourteen Shiba Inu dogs were presented to the Veterinary Medical Center at the University of Tokyo between June 1998 and June 2003. Among these, 46 dogs had glaucoma; the remaining 68 dogs were presented for routine vaccinations or heartworm testing and were used as a control population. Complete ophthalmic examination and gonioscopy were performed in all the dogs. PL and ICA were evaluated by gonioscopy and goniophotographs. Of the 46 dogs with glaucoma, 17 (37%) were affected bilaterally, and 29 (63%) were affected unilaterally. Of the 29 dogs with unilateral glaucoma, the ICA of the normotensive eye was slightly narrow in 2 eyes (7%), narrow in 12 eyes (41%), and closed in 15 eyes (52%). Among the normotensive control group, the ICA was open in 13 dogs (19%), slightly narrow in 29 (43%), narrow in 22 (32%), and closed in 4 (6%). In dogs with narrow and slightly narrow ICAs, the PLs were thickened, and in some cases, formed a solid sheet. The majority of dogs with glaucoma had changes in both ICA and PL. A narrow or slightly narrow ICA was also detected in the majority of normotensive control dogs. This study suggests that ICA narrowing and PL thickening is a common abnormality in Shiba Inu dogs in Japan and may predispose those dogs to glaucoma.     

Functional insulin‐like factor 3 (INSL3) hormone‐receptor system in the testes and spermatozoa of domestic ruminants and its potential as a predictor of sire fertility

Insulin‐like factor 3 (INSL3) is essential for fetal testis descent, and has been implicated in the testicular and sperm functions in adult males; however, similar functions in domestic ruminants remain largely unknown. This study investigated the functional INSL3 hormone‐receptor system in adult ruminant testes and spermatozoa, and explored its potential to diagnose the fertility of sires. Testes and spermatozoa were obtained from fertile bulls, rams and he‐goats, whereas subfertile testes and spermatozoa were obtained only from bulls. As expected, INSL3 was visualized in Leydig cells, while we clearly demonstrated that the functional receptor, relaxin family peptide receptor 2 (RXFP2), enabling INSL3 to bind was identified in testicular germ cells and in the sperm equatorial segment of bulls, rams and he‐goats. In comparison to fertile bulls, the percentage of INSL3‐ and RXFP2‐expressing cells and their expression levels per cell were significantly reduced in the testes of subfertile bulls. In addition, the population of INSL3‐binding spermatozoa was also significantly reduced in the semen of subfertile bulls. These results provide evidence for a functional INSL3 hormone‐receptor system operating in ruminant testes and spermatozoa, and its potential to predict subfertility in sires.     

Combined administration of ghrelin and GHRH synergistically stimulates GH release in Holstein preweaning calves

Ghrelin is a gut peptide which participates in growth regulation through its somatotropic, lipogenic and orexigenic effects. Synergism of ghrelin and growth hormone-releasing hormone (GHRH) on growth hormone (GH) secretion has been reported in humans and rats, but not in domestic animals in vivo. In this study, effects of a combination of ghrelin and GHRH on plasma GH and other metabolic parameters, and changes in plasma active and total ghrelin levels were studied in Holstein bull calves before and after weaning. Six calves were intravenously injected with vehicle (0.1% BSA-saline), ghrelin (1 microg/kg BW), GHRH (0.25 microg/kg BW) or a combination of ghrelin plus GHRH at the age of 5 weeks and 10 weeks (weaning at 6 weeks of age). Ghrelin stimulated GH release with similar potency as GHRH and their combined administration synergistically stimulated GH release in preweaning calves. After weaning, GH responses to ghrelin and GHRH became greater compared with the values of preweaning calves, but a synergistic effect of ghrelin and GHRH was not observed. The GH areas under the concentration curves for 2h post-injection were greater in weaned than in preweaning calves (P<0.05) if ghrelin or GHRH were injected alone, but were similar if ghrelin and GHRH were injected together. Basal plasma active and total ghrelin levels did not change around weaning, but transiently increased after ghrelin injection. Basal plasma insulin, glucose and non-esterified fatty acid levels were reduced after weaning, but no changes by treatments were observed. In conclusion, ghrelin and GHRH synergistically stimulated GH release in preweaning calves, but this effect was lost after weaning.     

Central administration of neuropeptide B, but not prolactin-releasing peptide, stimulates cortisol secretion in sheep

Two neuropeptides, neuropeptide B (NPB) and prolactin-releasing peptide (PrRP), have been suggested to play important roles in control of the hypothalamic-pituitary-adrenal (HPA) axis in rodents. The aim of the present study was to clarify the central actions of NPB or PrRP in sheep. Ovariectomized ewes were surgically implanted with a cannula directed to the lateral ventricle. They received intracerebroventricular (icv) administration of 400 mul of artificial cerebrospinal fluid, NPB (0.05, 0.5 or 5 nmol), PrRP (0.5, 5 or 50 nmol) or corticotropin-releasing hormone (CRH, 0.5 or 5 nmol) through the cannula, and blood samples were taken 30 and 0 min prior to and 15, 30, 60 and 90 min after the injection. Cortisol concentrations in plasma were determined by enzyme immunoassay. Administration of 0.5 nmol NPB resulted in a significant increase in the cortisol concentration compared with the vehicle control, whereas the cortisol concentration after lower or higher doses of NPB did not differ from the control value. Thus, an icv injection of NPB produced a bell-shaped dose-response of cortisol concentration. Administration of PrRP had no significant effect on the cortisol concentrations at any dose examined. Icv injection of CRH dose-dependently increased plasma cortisol concentrations. These results demonstrate that central NPB stimulates cortisol secretion, suggesting that this neuropeptide plays some roles in control of the HPA axis in sheep. On the other hand, unlike its role in rodents, PrRP is unlikely to be involved in control of the HPA axis in this species.     

Effect of suckling on the reproductive performance and metabolic status of obese Japanese black cattle during the early postpartum period

The aim of our study was to investigate the effects of suckling on reproductive performance and metabolic status of obese (mean body condition score of more than 4.0 on a scale of 1-5) maternal Japanese Black cows during early postpartum period. We used 7 postpartum Japanese Black cattle. Four cows were suckled ad libitum (suckled) until completion of their first artificial insemination (AI), while 3 cows were not suckled at all because they were separated from their calves immediately after parturition (non-suckled). Body weight and plasma concentrations of metabolites and hormones were measured from wk 1 to 9 postpartum. Ovarian activity was detected using plasma progesterone concentration, and all cows received their first AI after application of the Ovsynch protocol at approximately 4 months postpartum. Although body weights of non-suckled cows increased during experimental period (P<0.05), those of suckled cows remained unchanged. Plasma concentrations of glucose of non-suckled cows were higher at wk 2 postpartum (P<0.05) and their levels of non-esterified fatty acid tended to be lower at wk 1 and 2 postpartum compared with suckled cows (P<0.1); however, these differences between groups were not observed with progression of postpartum period. In addition, plasma insulin concentrations of non-suckled cows were higher than those of suckled cows during experimental period (P<0.05). During sampling period (wk 0 to 9 postpartum), onset of normal ovarian cycle was observed in all non-suckled and 2 of 4 suckled cows, and it was delayed in other 2 suckled cows compared with non-suckled cows; however, 3 suckled cows conceived at the first AI after application of the Ovsynch protocol; none of non-suckled cows conceived at this time. Overall, we suggest that suckling seems to reduce increase of body weight after parturition, although it does not improve obesity, and influences conception despite delay in resumption of normal ovarian cyclicity in obese Japanese Black cows.     

Epsilon-polylysine microparticle adjuvant drives cytokine production to Th1 profile

Epsilon-polylysine micro particles (SGEPL) and polyethyleneimine micro particles (SGPEI) were developed by the addition of a hydrophobic group and the immunological characterization of these micro particles and aluminum hydroxide (ALUM) was investigated. BALB/c mice were injected intraperitoneally with ovalbumin (OVA) as an antigen and SGEPL, SGPEI or ALUM as an adjuvant. The results showed that the mice injected with SGEPL produced a significant portion of anti-OVA antibody subclass IgG2a in the sera and suppressed interleukin (IL)-4 and IL-5, but enhanced IL-12 and interferon-gamma (IFN-gamma) from the spleen cells. Similar results relating to cytokines were also obtained, even without OVA. Direct stimulation with SGEPL to na?ve BALB/c mouse spleen cells induced IL-12 and IFN-gamma. Both spleen and purified B cells produced IgG1 and IgE after stimulation with IL-4 and the anti-CD40 monoclonal antibody. With the addition of SGEPL, the IgE production from the cells was suppressed as a result of enhanced IFN-gamma production. Furthermore, IgE production was also suppressed in the purified B cells without the influence of IFN-gamma or IL-12. Thus, we suggest SGEPL drives cytokine production to Th1 profile. It will be a novel promising adjuvant based on this viewpoint.