Ocular absorption and toxicity of methotrexate in the dog

The ocular absorption, elimination and toxicity of 12.5 mg of methotrexate in 0.5 cc administered subconjunctivally was studied in the dog. The purpose of this study was to establish ocular pharmacokinetics for treatment of intraocular tumours in dogs. Drug levels were measured in ocular compartments, and serum at 1,2,4,6,8 and 12 hours respectively. Tumoricidal concentrations of the drug were attained in the anterior chambers of the injected eyes up to and including six hours postinjection. Consistent signs of toxicity were anorexia, vomiting, weight loss and leukopenia. Local ocular toxicity following subconjunctival injection was minimal conjunctival hyperemia lasting up to 48 hours.     

Prevalence of canine coronavirus (CCoV) in dog in Japan: detection of CCoV RNA and retrospective serological analysis

We collected rectal swabs from dogs in Japan during 2011 to 2014, and canine coronavirus (CCoV) nucleocapsid gene was detected by RT-PCR. The relationship between CCoV infection and the manifestation of diarrhea symptoms was investigated, and a correlation was noted (df=1, χ²=8.90, P<0.005). The types of CCoV detected in samples from CCoV-infected dogs were CCoV-I in 88.9% and CCoV-II in 7.4%, respectively. We retrospectively investigated the seroprevalence of CCoV-I in dogs in Japan during 1998 to 2006. The sera were tested with a neutralizing antibody test. In the absence of CCoV-I laboratory strain, we used feline coronavirus (FCoV)-I that shares high sequence homology in the S protein with CCoV-I. 77.7% of the sera were positive for neutralizing anti-FCoV-I antibodies.     

The susceptibility of various spruce species to Rhizosphaera kalkhoffii and some cultural characteristics of the fungus in vitro

Temperature and pH requirements for mycelial growth and conidial germination were investigated for various Rhizosphaera kalkhoffii isolates. Conidial germination was also tested on spruce needles. Some factors of host susceptibility were determined using needle extract agar and buffer-capacity of needle sap.     

Measurement of erythrocyte carbonic anhydrase isozymes (CA-I and CA-II) in racehorses and riding horses

Equine carbonic anhydrase isozymes (CA-I and CA-II) were purified from erythrocytes by several column chromatography. Polyclonal anti-CA-I and anti-CA-II sera were produced in rabbits. Sensitive competitive enzyme-linked immunosorbent assays (ELISA) were established to determine the developmental changes in CA-I and CA-II levels in equine erythrocytes. Concentrations of CA-I and CA-II in erythrocytes from 150 clinically normal thoroughbreds (123 racehorses and 27 riding horses) were determined by ELISA. Mean (+/- SD) concentrations of CA-I and CA-II in racehorses were 1.70 +/- 0.48 and 0.94 +/- 0.13 mg/g hemoglobin (Hb), respectively. Mean concentrations of CA-I and CA-II in riding horses were 2.34 +/- 0.52 and 0.76 +/- 0.08 mg/g Hb, respectively. When the CA levels in racehorses and riding horses were compared, the CA-I level in riding horses was higher than that in racehorses (p=0.01). The CA-II level in racehorses was higher than that in riding horses (p=0.02). These data suggest that the levels of CA isozymes in erythrocytes of racehorses were influenced by chronic physical stress. The CA-I concentration in erythrocytes of 2-month-old horses was approximately 0.25 mg/g Hb. The CA-I level noticeably increased during the first year of life and approached normal adult levels by 2 years. The CA-II level decreased slightly with age, indicating different regulation of CA-I and CA-II expression during development.     

Comparison of cytokine mRNA expression in peripheral CD4+, CD8+ and γδ T cells between healthy Holstein and Japanese Black calves

Japanese Black (JB) calves are more susceptible to infectious diseases compared to Holstein (Hol) calves. To clarify the immunological differences between JB and Hol calves, expression of cytokine messenger RNA (mRNA) was examined using peripheral CD4⁺, CD8⁺ and γδ T cells. Healthy calves, 24 from each species, were examined. Blood samples were obtained from calves at 1 week, 1 month and 3 months old, eight calves for each age of each species. Peripheral blood mononuclear cells were stimulated with phytohemagglutin (PHA), and T cell subsets were isolated by positive selection using magnetic cell sorting (MACS). Levels of interlekin (IL)‐2, IL‐4, IL‐10 and interferon (IFN)‐γ mRNA in three T cell subsets were analyzed. WC1‐N1⁺ γδ T cell percentages were significantly lower in JB calves at 1 week and 1 month of age compared to Hol calves. In addition JB calves had significantly lower IL‐2, IL‐10 and IFN‐γ mRNA in WC1‐N1⁺ γδ T cells at 1 and 3 months of age, whereas there were no significant differences in cytokine mRNA of CD4⁺ and CD8⁺ cells between the two groups. Decreased cytokine mRNA and cell number of peripheral γδ T cells may affect negatively on the immune system of JB calves.     

Abscisic acid related compounds and lignans in prunes (Prunus domestica L.) and their oxygen radical absorbance capacity (ORAC)

Four new abscisic acid related compounds (1-4), together with (+)-abscisic acid (5), (+)-beta-D-glucopyranosyl abscisate (6), (6S,9R)-roseoside (7), and two lignan glucosides ((+)-pinoresinol mono-beta-D-glucopyranoside (8) and 3-(beta-D-glucopyranosyloxymethyl)-2- (4-hydroxy-3-methoxyphenyl)-5-(3-hydroxypropyl)-7-methoxy-(2R,3S)-dihydrobenzofuran (9)) were isolated from the antioxidative ethanol extract of prunes (Prunus domestica L.). The structures of 1-4 were elucidated on the basis of NMR and MS spectrometric data to be rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (1), rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid 3'-O-beta-d-glucopyranoside (2), rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (3), and rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxabicyclo[3,2,1]- oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (4). The antioxidant activities of these isolated compounds were evaluated on the basis of oxygen radical absorbance capacity (ORAC). The ORAC values of abscisic acid related compounds (1-7) were very low. Two lignans (8 and 9) were more effective antioxidants whose ORAC values were 1.09 and 2.33 micromol of Trolox equiv/micromol, respectively.     

Nuclear genome constitution and other characteristics of somatic hybrids between dihaploid Solanum acaule and tetraploid S. tuberosum

Eleven somatic hybrids (2n = 68 to 74) obtained between S. tuberosum ssp. tuberosum cv. Dejima (2n = 48) and ATDH-1 (2n = 24), an anther-culture-derived dihaploid of S. acaule (Yamada et al., 1997), were characterized by nuclear RFLP markers using 49 single-copy DNA probes distributed throughout the potato genome (2 to 6 probes per chromosome). One of the somatic hybrids, DA8-2, had 72 chromosomes and all the Dejima- and ATDH-1-specific markers (124 and 103 bands, respectively), suggesting the presence of a whole set of both parental chromosomes. The other somatic hybrids lost varying numbers of markers up to seventeen. The pattern of the loss of markers indicated the elimination of five chromosomes among four somatic hybrids. A nucleolar organizer region of chromosome 2 was often eliminated in the somatic hybrids. The somatic hybrids studied here had higher frequencies of multivalent formation than the S. tuberosum parent. They had reasonably good seed set when pollinated with S. tuberosum pollen. Hence, homoeologous recombination between S. acaule and S. tuberosum chromosomes is possible and useful traits from S. acaule may be transferred to the S. tuberosum gene pool. This revised version was published online in July 2006 with corrections to the Cover Date.     

Determination of Anthocyanins and Antioxidants in ‘Titanbicus’ Edible Flowers In Vitro and In Vivo

Plant Foods for Human Nutrition - Titanbicus (TB), a hybrid of Hibiscus moscheutos × H. coccineus (Medic.) Walt., has potential to be used as an edible flower. In this study, proximate...