Evaluation of the PATHFAST Chemiluminescent Enzyme Immunoassay for Measuring Progesterone in Whole Blood and Serum of Mares

Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone in whole blood and serum samples were measured by PATHFAST. In addition, concentrations of progesterone in serum samples measured by PATHFAST were compared with those measured by radioimmunoassay (RIA) and enzyme immunoassay (EIA). Using PATHFAST, the serum concentrations of progesterone in mares correlated highly with those of whole blood samples (r=0.9672, n=88). The serum concentrations of progesterone as measured by PATHFAST correlated well with RIA (r=0.9654, n=88) and EIA (r=0.9323, n=112). An abrupt decline in circulating progesterone in whole blood samples was observed within 2 hr (50%), followed by a gradual decline until 48 hr later. The results for progesterone in whole blood samples correlated highly with those in serum samples, and the declining pattern paralleled that of the serum samples. These results demonstrated that PATHFAST is useful in the equine clinic as an accurate diagnostic tool for rapid assay of progesterone within 26 min, using unextracted whole blood.     

Quantification of phosphatidic acid in foodstuffs using a thin-layer-chromatography-imaging technique

Apical application of lysophosphatidic acid (LPA), a growth-factor-like phospholipid, was shown to prevent or restore gastrointestinal (GI) disorders, such as diarrhea and stomach ulcer, in experimental animals. Because LPA is formed from phosphatidic acid (PA) by the activity of digestive phospholipase A(2), PA is a potential component for dietary treatment of such GI disorders. Here, we quantified PA contained in 38 foodstuffs and 3 herbs by a thin-layer-chromatography-imaging technique. Vegetables belonging to Brassicaceae, such as cabbage leaves (700 nmol/g of wet weight) and Japanese radish leaves (570 nmol/g), contained higher amounts of PA than other foodstuffs. Amounts of PA in fruits, cereals, and starchy root vegetables were below 300 nmol/g. Animal foodstuffs contained low amounts of PA (<60 nmol/g). Interestingly, leaves of Mallotus japonicas, a Japanese edible herb used for treatment of stomach ulcer, had the highest PA (1410 nmol/g) among those examined. The data shown here will be useful for the development of dietary treatment for a damaged GI tract.     

What is the optimal density of larval seeding in Acropora corals?

In situ larval seeding is a low-cost technique that is currently under development for the large-scale restoration of coral populations. One problem that still needs to be solved is the preparation of coral larvae for seeding, i.e., how many larvae are required to restore a certain area? In this study, we focused on the relationship between the numbers of larvae, settlers, and survivors for three?months post-settlement to determine the optimal larval seeding density. A comparison of three different larval densities (low, middle, and high) indicated that the number of settlers was proportional to the larval density, suggesting that settler density is determined by the number of larvae supplied. However, the survival rate of settlers on high-density plates was much lower than the corresponding rates on low- or middle-density plates during the first month after settlement. Moreover, most of the seeded corals had not survived on the low-density plates at three?months after settlement. Therefore, the middle larval density (i.e., 5000?larvae?m^?2) appears to be optimal for seeding on grid plates.     

Determinants of the daily rhythm of blood fluidity

Background  

Numerous processes in the living body exhibit daily rhythmicity. In this study, we characterized a daily rhythm of blood fluidity and identified its determinants.     

Current status of commercial Calibrachoa cultivars as assessed by morphology and other traits

To study the morphological diversity of the popular ornamental crop Calibrachoa, traded since the 1990s, floral and vegetative characters of 91 commercial cultivars were compared to those of natural species. Floral colours were classified according to the ISCC-NBS system and the CIE LAB system. The numerical data obtained by measuring 19 floral and vegetative characters was tested with principal component (PC) analysis. Floral colour classes of commercial cultivars were divided into 37 groups according the ISCC-NBS system, whereas those of natural species were divided into 9 groups. These tendencies were readily observable in the scatter diagrams of the a* vs. b* values and the C* vs. L* values. As the result of PC analysis, six PCs were obtained. For PC4, which negatively correlated with the degree of nocturnal corolla limb closure, the majority of natural species showed negative scores whereas the majority of cultivars had positive PC4 scores. In terms of PC2 (flower size), PC3 (growth form), PC5 (floral shape) and PC6 (leaf shape), natural species were dispersed in a range wider than the cultivars. These results suggest that Calibrachoa cultivars possess more highly variable floral colour than natural species, and breeding programmes to diversify the floral colour seem to have progressed rapidly. The tendency of natural species to close the corolla limb toward evening has been removed almost completely from the cultivars. The floral size, growth form, floral shape and leaf shape of the cultivars were rather uniform compared to those of natural species.     

Response changes with growth in the gustatory receptors of young yellowtails (Seriola quinqueradiata)

SUMMARY: Laboratory-hatched juvenile yellowtails (mean fork length, 9.3 cm) were reared for 2 months in two groups on different kinds of food, mysid and squid in one group and fish in the other, and the gustatory neural responses to seven chemicals (adenosine-5'-monophosphate, alanine, arginine, betaine, proline, tryptophan, and valine) were recorded in fish before and after the 2-month rearing period. Large variances in the responses to some chemicals were noticed in the juveniles before the rearing experiment. Mann–Whitney U -tests on the neural responses indicated significant changes in the magnitude of the responses to valine and some other chemicals relative to that for proline or tryptophan between the pre- and the two post-rearing groups. No significant differences in the response magnitude for the seven chemicals were observed for the two post-rearing groups suggesting that the response changes during the 2-month rearing might have been intrinsic and not due to specific food items in the diet. The dose–response relationship for some chemicals was also examined in the juveniles before and after the 2-month rearing. A slight lowering of threshold was noticed for alanine and valine after the rearing. Data on the responses of wild yellowtails were in support of the changing responsiveness of gustatory receptors during development of juvenile yellowtails.     

Leydig cell hyperplasia in an ENU-induced mutant mouse with germ cell depletion

Repro22 is an N-ethyl-N-nitrosourea (ENU)-induced mutation in mice showing depletion of both male and female germ cells. In the present study, we investigated the male phenotypes of the mutant mouse at the adult stage. The repro22/repro22 homozygous mice showed reduced body weights as well as markedly reduced testis weights. Histological examination of the testes at 4 and 10 months of age showed no germ cells in the seminiferous tubules of the affected testis while a number of Sertoli cells were observed in the tubules. In addition to the germ cell depletion, the testes of the affected mouse contained expanded intertubular spaces that were filled by Leydig cell-like interstitial cells. These interstitial cells were confirmed to be Leydig cells by immunohistochmical staining using anti-3beta-HSD antibody. The estimated number of Leydig cells in the affected testes at 10 months of age increased approximately 2 fold compared with those of normal testes. Furthermore, the plasma testosterone levels of the affected mice at 10 months of age were significantly higher than those of the normal mice. These findings indicated that the repro22/repro22 mouse developed hyperplasia of Leydig cells that was presumably caused by the absence of germ cells in the seminiferous tubules.     

Dynamics of lingual antimicrobial peptide,lactoferrin concentrations and lactoperoxidase activity in the milk of cows treated for clinical mastitis

The aim of the present study was to examine changes in innate immune factors in the milk of mastitic dairy cows treated with antibiotics. Cows in the antibiotics group (n = 13) were infused into the mammary gland with cefazolin on the sixth day after mastitis was diagnosed (the day of the mastitis diagnosis = day ?6). The control group (n = 12) was not treated. Milk samples were collected once every 2 days from days ?6 to 12 and somatic cell count (SCC), lingual antimicrobial peptide (LAP), and lactoferrin (LF) concentrations and lactoperoxidase (LPO) activity were measured. SCC and LF concentrations in the antibiotics group markedly decreased after the antibiotic treatment. When cows in the antibiotics group were divided according to SCC on day 0, LAP concentrations and LPO activity in cows with a lower SCC on day 0 (<5 × 10⁶ cell/mL) were significantly higher and lower than those in cows with a higher SCC, respectively. These results suggest that LF concentration decreased with decrease in SCC after treatment and that LAP concentration and LPO activity differed depending on the severity of mastitis. This is the first report to reveal the dynamics of innate immune factor in milk of cows treated for clinical mastitis.     

Patterns of shoot mortality in Betula platyphylla in northern Japan

To understand the development of crown structure in Betula platyphylla Sukatch., mortality patterns of long shoots were analyzed quantitatively. We selected 25 saplings growing under various light conditions and measured the relative photosynthetically active radiation (rPAR) at, and the three-dimensional position of, first-order branches. A long shoot was assigned "no buds" (NB) status if it lacked buds at the end of the growing season, including at the tips of short shoots. A long shoot was classified as dead if it was NB and all the offspring long shoots issuing from it were NB. The probability that a leafy long shoot (a current-year long shoot with leaves or an older long shoot with short shoots with leaves) would become NB by the end of the season was positively dependent on shoot age and branch age, and negatively dependent on shoot length, centripetal shoot order, branch height and rPAR at the branch. Randomization tests revealed that shoots became NB and dead in clusters of connected shoots. In particular, shoot clusters originating from 3-year-old shoots were more likely to die than expected if each shoot was assumed to become NB regardless of the connection. Stepwise logistic regression revealed that the maximum rPAR within the crown of an individual tree had a significant effect on the mortality rate of 3-year-old shoot clusters, together with the rPAR at the level of the branch and other structural entities. Correlative inhibition is an important mechanism for determining shoot mortality patterns.     

Effects of acidic fog and ozone on the growth and physiological functions of Fagus crenata saplings

The effects of simulated acid fog (SAF) and ozone (O₃) stress on the growth and physiology of beech (Fagus crenata) saplings were investigated. Three-year-old beech saplings were exposed to SAFs of pH 3 and pH 5 (control) during May 2007 to July 2008. In each SAF treatment group, half of the saplings were exposed to 60 ppb of O₃ during September 2007 to July 2008. In comparison to the control saplings, those from the pH 3 treatment had lower total plant biomasses, epicuticular wax amounts, Ca²⁺ concentrations in their leaves, and lower starch concentrations in their leaves and roots. The effect of O₃ was significant only for the starch concentration in the roots, but the O₃ exposure also negatively affected the growth and physiology of beech saplings. Results show that acid fog exerts various severe effects, and that both chronic acid fog and O₃ exposure suppressed the physiological functions of beech saplings.