Regional blood flow distribution from the proximal arterial cannula during veno-arterial extracorporeal membrane oxygenation in neonatal dog.

Extracorporeal membrane oxygenation (ECMO) is frequently used for treatment of patients with severe hypoxemia due to life-threatening respiratory failure. Due to this hypoxemia, the myocardium of these patients is insufficiently provided with oxygen, and consequently their cardiac function commonly deteriorates. But veno-arterial (V-A) ECMO provides oxygenated blood to the coronary arteries from ECMO circuit insufficiently. To increase the coronary blood flow distributed from ECMO, we placed the arterial cannula 1 cm above the aortic valve and evaluated the regional blood from the proximal arterial cannula in comparison with the distal cannula. Eight neonatal dogs weighting 1.8-2.5 kg were supported by V-A ECMO. The regional blood flow from the arterial cannula was measured by injection of colored microspheres into ECMO circuit. The site of the arterial cannula was changed under fluoroscopy. The bypass flow was maintained at either 50 or 100 ml/min/kg. We found that the coronary blood flow distributed from the proximal arterial cannula was significantly higher than that from the distal cannula. The proximal arterial cannula appears necessary to provide sufficient oxygenated blood to the coronary circulation during V-A ECMO. Therefore, it is expected that the increased cardiac function may improved, and that the survival rate of the patients with retarded cardiac function due to severe hypoxemia may increase by proximal placement of the arterial cannula during V-A ECMO.     

The secretory pattern and source of immunoreactive prolactin in pregnant African (Loxodonta africana) and Asian (Elephas maximus) elephants

The objective of the present study was to define the secretion of prolactin (PRL) in pregnant African and Asian elephants. Levels of immunoreactive (ir-) PRL in serum and placental homogenates were measured by a heterologous radioimmunoassay (RIA) based on an ovine and human RIA system, and the localization of ir-PRL in the placenta was detected by immunohistochemistry using anti-human PRL. Circulating ir-PRL clearly showed a biphasic pattern during pregnancy in African and Asian elephants. Serum levels of ir-PRL started to increase from the 4 - 6th month of gestation and reached the first peak level around the 11-14th month. A second peak of circulating ir-PRL levels was observed around the 18-20th month of gestation followed by an abrupt decline after parturition. In contrast, in a case of abortion of an African elephant, the second peak of ir-PRL was not observed, and the levels remained low for about four months until parturition. The weight of the fetus delivered at the 17th month of gestation was 23.5 kg, which was quite small compared with normal fetuses in previous reports. Ir-PRL was detected in placental homogenates, and immunolocalization was observed in trophoblasts in both the African and Asian elephants, indicating that the placenta is the source of ir-PRL during pregnancy in elephants. The present results clearly demonstrated that circulating ir-PRL shows a biphasic pattern during normal pregnancy and that the placenta appears to be an important source of circulating ir-PRL during pregnancy in both African and Asian elephants.     

Analysis of cartilage oligomeric matrix protein (COMP) degradation and synthesis in equine joint disease

REASONS FOR PERFORMING STUDY: Cartilage oligomeric matrix protein (COMP) is abundant within cartilage; its turnover and/or degradation have been investigated in various equine joint diseases and it has been suggested that COMP fragmentation might be useful for monitoring such conditions. OBJECTIVES: To determine whether COMP metabolism is compromised in equine osteoarthritis (OA) and whether COMP degradation is a useful joint marker representing cartilage destruction. HYPOTHESIS: A monoclonal antibody (mAb) with a higher affinity for degraded COMP allows discrimination of diseased joints by quantifying COMP levels and fragmentation. METHODS: A mAb (clone14G4) was generated against equine cartilage COMP. The NH2-terminal sequence of enzyme-cut COMP fragments recognised by 14G4 was determined, as was the efficiency of binding to COMP (using a generated COMP peptide). COMP concentration and fragmentation were analysed in synovial fluid (SF) from normal horses and those with OA. RESULTS: The mAb 14G4 had a higher affinity for the smaller fragments of equine COMP, compared with a mAb (clone 12C4) generated against human COMP. The 14G4 epitope was identified as between C134 and F147. The COMP values in OA (mean +/- s.d. 205.8 +/- 90.9 microg/ml) were significantly higher than in the normal (133.1 +/- 31.5 microg/ml) SF. On the immunoblots of OA sample, the proportions of intact COMP were significantly lower, while smaller fragments ranging from 75 to 290 kDa were higher compared with the normal SF. CONCLUSIONS AND POTENTIAL RELEVANCE: The mAb 14G4 reliably detects COMP degradation as well as synthesis, and fragmentation analysis combined with quantification in SF could be useful to study equine OA.     

The g.841G>C SNP of FASN gene is associated with fatty acid composition in beef cattle

The objective of the current study is to evaluate the association between fatty acid composition and fatty acid synthase gene polymorphisms as responsible mutations. For this purpose, we selected seven previously reported single nucleotide polymorphisms (SNPs) in FASN gene, including one within promoter region (g.841G>C) and six non‐synonymous SNPs (g.8805C>T, g.13126C>T, g.15532A>C, g.16024A>G, g.16039C>T, g.17924A>G), and genotyped them in Japanese Black cattle. Genotyping results revealed that g.8805 C>T and g.17924 A>G were monomorphic loci. Genome‐wide association analysis including the other five SNPs revealed that only g.841G>C showed significant associations with the percentages of C14:0, C14:1, C16:1 and C18:1 at 5% genome‐wide significance level. In order to further evaluate the effect, we genotyped g.841G>C using additional three populations, including two Japanese Black populations and a Holstein cattle population. g.16024A>G was also genotyped and included in the analysis because it has been reported to be associated with fatty acid composition in Japanese Black cattle. In the result of analysis of variance, g.841G>C showed stronger effects on fatty acid percentage than those of g.16024A>G in all populations. These results suggested that g.841G>C would be a responsible mutation for fatty acid composition and contribute to production of high‐grade beef as a selection marker in beef cattle.     

The effects of growth and disease in serum keratan sulfate concentration in dogs

The purpose of this study is to investigate keratan sulfate (KS) concentration in the serum of puppies and the effects of age, body weight, breed and diseases. Serum samples from six neonatal dogs (4 Beagles, 2 Labrador Retrievers), and from 127 adult dogs with various diseases were collected at a Teaching Animal Hospital. Canine serum KS concentration was measured by inhibition enzyme-linked immunosorbent assay (ELISA). Samples from puppies were evaluated for growth-related changes, and samples from patients were evaluated for age, body weight, breed and disease-related changes. Serum KS concentration was high in puppies from birth to 4 months of age. KS values started to decrease from 4 months to 9 months of age, and then gradually reached to the plateau. Though in the small sample, mean KS concentration in a Labrador Retriever was higher than in Beagles during the first 10 months. The values of serum KS showed body weight-related increase within retrievers among teaching hospital population and there was significant increase in body weight-related change. Cartilage metabolism is high in canine immature joint and that activity continues for 5 months, and that higher in Labrador Retrievers rather than in Beagles. There was no effect from other factors, including age, body weight, breed and disease in all patients. Serum KS concentration of Retrievers is higher than Beagles, and that value increased with gain of body weight. We suggest that Retriever have higher cartilage metabolism with growth or ageing.     

Age-related Histological Findings in the Pineal Gland of Crl:CD(SD) Rats

To provide background data as the pathologic basis, the pineal glands of 190 male and 193 female Crl:CD(SD) rats at ages of 0–7, 51–58, 70–85 and 111 weeks were examined histologically in this study. Mineralization and fibrosis were common findings in the aged rats, whereas they were rarely found in the young ones; mineralization was present in 7, 44, 67 and 79% of males and in 0, 32, 67 and 79% in females, and fibrosis was present in 0, 29, 48 and 44% of males and 0, 18, 40 and 35% of females at ages of 0–7, 51–58, 70–85 and 111 weeks, respectively. Striated muscle fiber appeared regularly in the fibrosis region from 51–58 weeks of age when fibrosis increased, while the origin of this fiber remained unclear. Vacuolation of pineal cells also increased with age in both sexes, though the total incidence was low. There was a low incidence of lymphocytic infiltration in both sexes, but this was not related to age.     

Horizontal transmission and phylogenetic analysis of bovine leukemia virus in two districts of Miyazaki,Japan

Horizontal transmission is recognized as a major infection route for bovine leukemia virus (BLV), and cattle with high viral loads are considered to be a major infectious source in a herd. However, a correlation between viral loads and the risk of infection has been insufficient to use as a foundation for BLV control strategies. In this report, we examined the epidemiology of BLV infection and the infectious source in a local area. In 2013–2014, BLV infection was investigated in 1,823 cattle from 117 farms in two adjacent districts, Miyazaki, Japan. Seropositive samples for BLV were detected with 88 cattle and in 14 farms. Phylogenetic analysis revealed that 94% of the isolates clustered into genotype I and the remaining isolate into genotype III. Among genotype I, genetically distinct strains were spread at each farm, and cattle infected with less than 3 copies/100 cells did not transmit BLV to other cattle for more than thirty months. This is the first report of concrete data of viral load in relation to viral horizontal transmission under the field condition. The data facilitate farmers and veterinarians understanding the status of BLV infected cattle. This research contributes to BLV infection control and the development of effective BLV eradication programs.     

Comparison of Ammonium Deposition Flux at Roadside and at an Agricultural Area for Long-Term Monitoring: Emission of Ammonia from Vehicles

To understand the effect of the NH₃ emissions from vehicles, the NH₄ ⁺ bulk deposition and concentrations of gaseous NH₃ and particulate NH₄ ⁺ were measured at two sites in Saitama prefecture, Japan, one at the roadside of an arterial road (Kounosu), and the other in an agricultural area (Kisai). We observed that the NH₄ ⁺ bulk deposition and NH₃ concentration were significantly increased in Kounosu; the effect of NH₃ emissions from vehicles was confirmed. NOx was a primary pollutant mainly emitted by vehicle exhaust in the vicinity of arterial roads, and the relationship between NO _x and sum of gaseous NH₃ and particulate NH₄ ⁺ (T-NH₄ ⁺) was examined. The [T-NH₄ ⁺/NO _x ] ratios in Kounosu ranged from 0.08 to 0.23 (average 0.15), and in Kisai they ranged from 0.16 to 0.67 (average 0.39). In Kounosu, the effect of vehicle exhaust was great, but seasonal variation was small. The contribution ratio of NH₃ from vehicle emissions increased at low temperatures, while that from the non-vehicle emissions increased at high temperatures.     

Comparison of the glucosinolate-myrosinase systems among daikon (Raphanus sativus, Japanese white radish) varieties

Myrosinase is a cytosolic plant enzyme present in daikon ( Raphanus sativus, Japanese white radish) roots that hydrolyzes 4-methylthio-3-butenyl glucosinolate (MTBGLS) into the natural pungent agent 4-methylthio-3-butenyl isothiocyanate (MTBITC), which possesses antimicrobial, antimutagenic, and anticarcinogenic properties. The concentration of MTBGLS, myrosinase activity, and production of MTBITC in seven daikon varieties (one conventional and six heirlooms) were determined to rank the activity of the glucosinolate-myrosinase system and identify critical factors influencing the production of MTBITC. The six heirloom varieties produced 2.0-11.5 times higher levels of MTBITC as compared to the conventional variety, Aokubi, which is consumed by the present Japanese population. The myrosinase was located exclusively in the outer epidermal layer in Aokubi, and MTBGLS was widely distributed throughout the root tissue. Although the skin is a potentially rich source of myrosinase in Aokubi, the skin is usually peeled off in the current practice of preparing daikon for cooking. New practices are therefore proposed for the preparation of daikon tubers that eliminate the peeling of the skin to avoid removing the enzyme needed to convert MTBGLS to the health-beneficial MTBITC. It is also concluded that the consumption of heirloom daikon varieties in addition to changes in food preparation will optimize the health benefits of daikon.