Basal levels and GnRH-induced responses of peripheral testosterone and estrogen in Holstein bulls with poor semen quality

The present study investigated the basal levels and GnRH-induced responses of peripheral testosterone and estrogen in Holstein bulls with poor semen quality. On the basis of semen parameters, bulls (n=5) having poor semen quality were selected as experimental bulls, and good semen quality bulls (n=4) were used as control bulls. Both groups were treated intramuscularly once with GnRH (250 μg of fertirelin acetate). Blood samples were collected at -1 day (d), -30 min and 0 h (treatment) followed by every 30 min for 5 h and 1, 3 and 5 d post-GnRH treatment (PGT), and LH, testosterone and estradiol-17β (E(2)) concentrations were measured. The pretreatment concentrations were used as basal levels. The percentage increments based on the 0-h levels were calculated per bull for each sampling time until 5 h PGT, and differences were compared between the experimental and control groups. The PGT concentrations of testosterone and basal and PGT concentrations of E(2) were significantly lower in the experimental group. The testosterone increment in the experimental group was delayed and significantly lower from 1 to 5 h PGT than those in the control group. It can be suggested that bulls with poor semen quality have delayed and lower GnRH-induced testosterone response and may also have lower estrogen levels.     

DNA methylation status of bovine blastocyst embryos obtained from various procedures

DNA methylation is an important factor for the regulation of gene expression in early embryos. It is well known that the satellite I sequence is more heavily methylated in bovine somatic cell nuclear transfer (NT-SC) embryos than in embryos derived from in vitro fertilization (IVF). However, the methylation status of bovine embryos obtained by other procedures is not well known. To clarify DNA methylation levels of bovine embryos obtained from various procedures, we examined satellite I sequences in bovine blastocyst (BC) embryos derived from NT-SC, NT using embryonic blastomeres (NT-EM), in vivo (Vivo), IVF and parthenogenetic treatment (PA). Furthermore, in order to evaluate the efficacy of DNA demethylation by the NT procedure, we determined the DNA methylation levels in bovine embryos in which NT was recapitulated (Re-NT). Although the DNA methylation levels in the NT-SC embryos were higher than those in the other embryos, the NT-EM embryos exhibited lower DNA methylation levels. The satellite I sequence in the NT-SC embryos was more demethylated than that in the donor cells. Although the DNA methylation level in the individual NT-SC embryos showed variation, the full-term developmental efficacy of these embryos were not different. These findings suggest that the methylation level of the satellite I sequence at the BC stage is not related to the abnormalities of bovine embryos produced by NT-SC. There was no difference in methylation levels between Re-NT and NT-SC embryos. Our results indicated that the DNA methylation status differed among embryos produced by various methods and that at least some of the demethylation of the donor cell genome occurred in the recipient cytoplast after NT-SC, but the demethylation ability of the NT procedure was noted in the first NT but not in the second NT.     

Differences of tensile strength distributions between mechanically high-grade and low-grade Japanese larch lumber III: effect of knot restriction on the strength of lumber

It is well known that the presence of knots in structural lumber is one of the most important strengthreducing factors. For practical purposes, visual grading including knot restriction is an effective method for nondestructive evaluation of strength. Edge knot restriction for not only visually graded lumbers but also mechanically graded lumbers is specified in the Japanese agricultural standards for glued laminated lumber. We conducted experimental studies on differences of tensile strength distributions between mechanically high-grade and low-grade Japanese larch (Larix kaempferi, carriere) lumbers daily used for manufacturing glued laminated timbers in Nagano, Japan. We then examined the additional visual grading of mechanically graded lumbers for nondestructive evaluation. We visually graded the prepared mechanically graded lumber by focusing on the knots' area ratio of grouped knots. We confirmed that the higher visual grade related to the stronger tensile strength, similar to our present knowledge; but the effects of knot restriction were reduced when the length of the lumber increased in view of nonparametric 5th percentiles of tensile strength. The differences in the strength/elasticity ratio between mechanically high-grade and low-grade lumber were negligible. It was clear that the length effect on the ratio in visually graded high-grade lumber was smaller than that of visually graded low-grade lumber. It was thus concluded that knot restriction should have little effect on the tensile strength of mechanically graded lumber.     

Microdialysis measurement of intracerebral somatostatin in the goat

A microdialysis sampling technique for the intracerebral measurement of somatostatin (SS) in extracellular fluid was examined in the goat. The microdialysis probe (70-mm shaft, 0.5 mm outer diameter) contained at its tip a 4-mm length of copolymer dialysis membrane (20 kDa cut-off). Artificial cerebrospinal fluid (artificial CSF) was pumped through the probe tip at a rate of 4 μl/min with a battery-driven syringe pump, and effluent fractions of dialysate (120 μl) were collected every 30 min. An in vitro recovery test showed that changes in the SS concentration in dialysate were highly correlated (r = 0.95, P < 0.01) with those in the external medium, and the relative recovery averaged 2.0%. As a validation for in vivo microdialysis, trails were conducted with conscious behaving goats wherein the inflow dialysate was changed transiently from artificial CSF with low potassium (2.5 mM) to a solution of 300 mM KCl. Potassium-induced depolarization around the probe tip located in the preoptic area and in the hypothalamus induced an increase in SS concentrations in dialysate at each location. In the most remarkable response, the concentrations of SS were increased 6-fold and 11-fold in the first and second 30-min fractions, respectively, compared with prepotassium concentrations. These results suggest that intracerebral SS levels in extracellular fluid could be estimated from conscious behaving goats by the use of our intracerebral microdialysis system.     

Influence of coating properties on paper-to-paper friction of coated paper

Coated paper-to-coated paper friction properties were examined in relation to printing runnability difficulties like erroneous double feeding of paper sheets. Higher ratios of precipitated calcium carbonate (PCC) to clay in mixed pigment coatings resulted in higher static and kinetic coefficients of friction (COFs). Microroughness in the order of pigment particle size is considered to relate to COF, because cube-shaped particles of PCC resist sliding. Calendering decreased COF at larger amounts of PCC, but did not change COF of the sole clay formulation at all. Addition of ground calcium carbonate (GCC) decreased COF. The rate of decrease in kinetic COF with increasing number of sliding for the GCC-rich formulation was higher than that for the PCC-rich formulation, presumably because protruding parts, characteristic of the GCC-rich formulation, on the surface were selectively flattened. Addition of styrene-butadiene (SB)-latex up to 14pph decreased COF, but static COF had the highest value at 18pph. The antislip property (as a rubber) of SB-latex developed only in the static mode. Among lubricants formulated, the wax type decreased COF the most remarkably with more effect on kinetic COF than on static COF. Part of this report was presented at the 11th International Printing and Graphic Arts Conference, Bordeaux, France, October 2002     

Effective sampling method for estimating bending strength distribution of Japanese larch square-sawn timber

Information on the strength distribution of timbers and other wood products seems to have become more important for users and producers after revision of the Japan architectural standard in 1998, which emphasizes the performance requirements of structures. Because there is no way other than expensive destructive tests to collect strength data, many researchers have proposed many inspecting methods for predicting strength by nondestructive evaluation. The most popular method for structural timber is the mechanical grading method based on the relation between Young's modulus (E) and strength () with some linear regression models. On the other hand, it is well known that the proof loading test is superior for obtaining information on the lower tail of distribution. If the E distribution of the objective timbers is known approximately, selecting timbers nearest to the projected E values saves timbers for destructive tests. We examined the alternative sampling method using the reported e- data sets of Japanese larch square-sawn timber. The simulated results showed that the estimated lower tail of the bending strength distribution by the alternative method was a better approximation of the experimental distribution than that derived from the conventional linear regression model.Part of this paper was presented at the 47th Annual Meeting of the Japan Wood Research Society, Kochi, April 1997     

Light-induced off-flavor development in cloudy apple juice

Cloudy apple juice has been found to develop off-flavors during storage in daylight. The development of off-flavors and volatile compounds was monitored in reconstituted juice prepared from 'Golden Delicious' and 'Fuji' apple concentrates stored in glass bottles under fluorescent light (3000 lx, 8 degrees C). A strong metallic off-flavor was formed by photooxidation. A major contributor to the off-flavor was identified as 1-octen-3-one by gas chromatography-olfactometry. In addition, six volatile compounds, pentanal, 2-methyl-1-penten-3-one, hexanal, (E)-2-heptenal, 6-methyl-5-hepten-2-one, and (E)-2-octenal, increased significantly after light exposure and could contribute to the off-flavor. Except for pentanal and hexanal, these volatiles were found only after light exposure. Higher levels of volatiles were observed in juice from 'Golden Delicious' apples than in juice from 'Fuji' apples, and this difference was consistent with higher levels of suspended solids. When the suspended solids were removed by centrifugation, the development of volatiles on exposure to light was reduced significantly.     

Characteristics of prolactin-releasing response to salsolinol (SAL) and thyrotropin-releasing hormone (TRH) in ruminants

The secretion of prolactin (PRL) is stimulated by thyrotropin-releasing hormone (TRH), and inhibited by dopamine (DA). However, we have recently demonstrated that salsolinol (SAL), a DA-derived endogenous compound, is able to stimulate the release of PRL in ruminants. The aims of the present study were to compare the characteristics of the PRL-releasing response to SAL and TRH, and examine the relation between the effects that SAL and DA exert on the secretion of PRL in ruminants in vivo and in vitro. Three consecutive intravenous (i.v.) injections of SAL (5 mg/kg body weight (b.w.): 19.2 μmol/kg b.w.) or TRH (1 μg/kg b.w.: 2.8 nmol/kg b.w.) at 2-h intervals increased plasma PRL levels after each injection in goats (P < 0.05); however, the responses to SAL were different from those to TRH. There were no significant differences in each peak value between the groups. The rate of decrease in PRL levels following the peak was attenuated in SAL-treated compare to TRH-treated animals (P < 0.05). PRL-releasing responses to SAL were similar to those to sulpiride (a DA receptor antagonist, 0.1 mg/kg b.w.: 293.3 nmol/kg b.w.). In cultured bovine anterior pituitary (AP) cells, TRH (10⁻⁸ M) significantly increased the release of PRL following both 15- and 30-min incubation periods (P < 0.05), but SAL (10⁻⁶ M) did not increase the release during the same periods. DA (10⁻⁶ M) completely blocked the TRH-induced release of PRL for a 2-h incubation period in the AP cells (P < 0.05). Sulpiride (10⁻⁶ M) reversed this inhibitory effect but SAL (10⁻⁶ M) did not have any influence on the action of DA. These results show that the mechanism(s) by which SAL releases PRL is different from the mechanism of action of TRH. Furthermore, they also show that the secretion of PRL is under the inhibitory control of DA, and SAL does not antagonize the DA receptor's action.     

Superficial necrolytic dermatitis associated with extrapancreatic glucagonoma in a dog

An 11-year-old Shih Tzu presented with crusting and erythema, mainly on the abdomen and the root of the tail. Based on histopathological findings, blood examinations and necropsy findings, the condition was diagnosed as superficial necrolytic dermatitis associated with a glucagon-secreting extrapancreatic neuroendocrine tumour. Gross necropsy revealed tumour invasion into the spleen, liver, adrenal glands and mesenteric lymph nodes. Immunohistochemical analysis of the neoplastic cells revealed that the tumour was a glucagonoma, consistent with earlier findings of persistent glucagonaemia and hypoaminoacidaemia.     

Dysfunction of erythropoietin-producing interstitial cells in the kidneys of ICR-derived glomerulonephritis (ICGN) mice

Anemia is a major secondary symptom in chronic renal disorder (CRD), but the precise cause of insufficient production of erythropoietin (EPO) remains unclear owing to the controversial localization of EPO-producing cells in the kidneys. The ICR-derived glomerulonephritis (ICGN) mouse, a new hereditary nephrotic mouse, is an appropriate model of anemia associated with CRD. By using an amplified in situ hybridization technique, we detected and counted the renal EPO-producing cells under both normoxic and hypoxic conditions. The expression levels of renal EPO mRNA were quantified and oxygen gradients were also assessed immunohistochemically. Amplified in situ hybridization clarified that EPO-producing cells were peritubular interstitial cells in the middle region of renal cortex in both ICR and ICGN mice. Hypoxia (7% O2) induced low oxygen tension in proximal tubular epithelial cells of renal cortex, and increased the expression of EPO mRNA and the number of EPO-producing cells in both ICR and ICGN mice. However, hypoxia did not increase the serum EPO levels in ICGN mice. The ICGN mouse is a good model for anemia associated with CRD, and the suppression of EPO protein production in the renal EPO-producing cells is considered to be a potential cause of anemia associated with CRD.