Preliminary system of rapid analysis of blood retinol level in cattle

Control of blood retinol levels in cattle during fattening is important in the production of marbled beef. However, it is difficult to easily measure the blood retinol concentration in the field. In this study, we attempted to develop an analysis method that does not require blood cell separation and uses a compact fluorescence analyzer that can be carried around as a preliminary system for measuring blood retinol concentration in the field. This system was used to monitor blood retinol levels in 12 fattening cattle (14 to 27 months old) and demonstrated a strong correlation (r = 0.78) with the results obtained by the standard high-performance liquid chromatography (HPLC) method. Stronger correlations (r = 0.87) were obtained until the cattle were 24 months of age. These results suggest that higher correlations can be expected to be obtained by improving the robustness of the extraction system. Refinements for practical use need to be considered, but whole blood extraction and the vitamin A analyzer that was developed show potential to be used for on-farm monitoring of retinol levels.     

Evaluation of the Autonomic Nervous System in a Canine Model of Chronic Embolic Pulmonary Hypertension

Veterinary Research Communications - Sildenafil improves autonomic dysfunction caused by pulmonary hypertension (PH) in humans, but its effect is unknown in dogs with PH. This prospective study...     

Molecular biological and immunohistological characterization of canine dermal papilla cells and the evaluation of culture conditions

The dermal papilla (DP) plays pivotal roles in hair follicle morphogenesis and cycling. However, our understanding of the biology of the canine DP is extremely limited. The aim of this study was to elucidate molecular biological and immunohistochemical characteristics of canine DP cells and determine appropriate conditions for in vitro expansion. Histological investigation revealed that the canine DP expressed biomarkers of human and rodent DP, including alkaline phosphatase (ALP) and versican. When microdissected, canine DP, but not fibroblasts, strongly expressed the DP-related genes for alkaline phosphatase, Wnt inhibitory factor 1 and lymphoid enhancer-binding factor 1, confirming successful isolation. The growth rate of isolated canine DP cells was moderate in conventional culture conditions for rodent and human DP; however, AmnioMAX-C100 complete medium allowed more efficient cultivation. Dermal papilla marker gene expression was maintained in early passage cultured DP cells, but gradually lost after the third passage. Approaches to mimic the in vivo DP environment in culture, such as supplementation of keratinocyte-conditioned medium or use of extracellular matrix-coated dishes, moderately ameliorated loss of DP gene expression in canine DP cells. It is possible that constituent factors in AmnioMAX may influence culture. These findings suggested that further refinements of culture conditions may enable DP cell expansion without impairing intrinsic properties and, importantly, demonstrated that AmnioMAX-cultured early passage canine DP cells partly maintained the biological characteristics of in vivo canine DP cells. This study provides crucial information necessary for further optimization of culture conditions of canine DP.     

Earthquake source fault beneath Tokyo

Devastating earthquakes occur on a megathrust fault that underlies the Tokyo metropolitan region. We identify this fault with use of deep seismic reflection profiling to be the upper surface of the Philippine Sea plate. The depth to the top of this plate, 4 to 26 kilometers, is much shallower than previous estimates based on the distribution of seismicity. This shallower plate geometry changes the location of maximum finite slip of the 1923 Kanto earthquake and will affect estimations of strong ground motion for seismic hazards analysis within the Tokyo region.     

Intramolecular carbon isotope distribution of acetic acid in vinegar

Compound-specific carbon isotope analysis of acetic acid is useful for origin discrimination and quality control of vinegar. Intramolecular carbon isotope distributions, which are each carbon isotope ratios of the methyl and carboxyl carbons in the acetic acid molecule, may be required to obtain more detailed information to discriminate such origin. In this study, improved gas chromatography-pyrolysis-gas chromatography-combustion-isotope ratio mass spectrometry (GC-Py-GC-C-IRMS) combined with headspace solid-phase microextraction (HS-SPME) was used to measure the intramolecular carbon isotope distributions of acetic acid in 14 Japanese vinegars. The results demonstrated that the methyl carbons of acetic acid molecules in vinegars produced from plants were mostly isotopically depleted in (13)C relative to the carboxyl carbon. Moreover, isotopic differences (δ(13)C(carboxyl) - δ(13)C(methyl)) had a wide range from -0.3 to 18.2‰, and these values differed among botanical origins, C3, C4, and CAM plants.     

High-speed atomic force microscopy reveals rotary catalysis of rotorless F₁-ATPase

F(1) is an adenosine triphosphate (ATP)-driven motor in which three torque-generating β subunits in the α(3)β(3) stator ring sequentially undergo conformational changes upon ATP hydrolysis to rotate the central shaft γ unidirectionally. Although extensive experimental and theoretical work has been done, the structural basis of cooperative torque generation to realize the unidirectional rotation remains elusive. We used high-speed atomic force microscopy to show that the rotorless F(1) still "rotates"; in the isolated α(3)β(3) stator ring, the three β subunits cyclically propagate conformational states in the counterclockwise direction, similar to the rotary shaft rotation in F(1). The structural basis of unidirectionality is programmed in the stator ring. These findings have implications for cooperative interplay between subunits in other hexameric ATPases.     

METABOLITE PROFILING OF SHOOT EXTRACTS,ROOT EXTRACTS,AND ROOT EXUDATES OF RICE PLANT UNDER PHOSPHORUS DEFICIENCY

We applied a metabolite profiling technique to investigate root exudates under phosphorus (P) deficiency. Oryza sativa was grown in culture solution containing three P levels (0, 1, and 8 mg P L^?1). Shoot extracts, root extracts, and root exudates were obtained from 18 and 23-day-old plants and their metabolites were determined by capillary electrophoresis/time-of-flight mass spectrometry. Eighty, 90, and 65 metabolites were identified in shoot extracts, root extracts, and root exudates, respectively. Sixty-three to eighty-four percent of the metabolites were exuded to the rhizosphere. More than 33% of the metabolites in the root exudates showed higher concentration at low P than at high P. On the other hand, only 14% of the metabolites in the root extracts showed lower concentration at low P than at high P. These results suggest that rice roots actively release many metabolites in response to P deficiency.     

Improvement of simazine degradation by inoculation of corn and soybean plants with rhizobacteria

We isolated the strains of aerobic bacteria, that were able to use the simazine (2-chloro4,6-bis(ethylamino)-s-triazine) herbicide as a sole source of nitrogen, from the roots of corn plants and the nodules of soybean plants. Partial base sequences of 16S rRNA genes and physiological characteristics of the representative isolates revealed that they belonged to Agrobacterium radiobacter and Bradyrhizobium japonicum, respectively. Inoculation of corn plants with the isolates of A. radiobacter resulted in a faster decrease in the amount of simazine in vermiculite soil. Neither bacteria nor corn plants alone were able to promote the decrease in the amount of simazine. Soybean plants modulated by the isolates of B. japonicum were more sensitive to simazine in a hydroponic medium compared to non-nodulated soybeans, though a significant decrease in the amount of simazine in the media was observed. The results obtained in the present study indicate that the rhizosphere of corn and soybean plants containing root nodules is an appropriate habitat for simazine-degrading bacteria to degrade the herbicide.     

Epidermal structure created by canine hair follicle keratinocytes enriched with bulge cells in a three‐dimensional skin equivalent model in vitro: implications for regenerative therapy of canine epidermis

Background – Keratinocytes in the hair follicle bulge region have a high proliferative capacity, with characteristics of epithelial stem cells. This cell population might thus be an ideal source for generating the interfollicular epidermis in a canine skin equivalent. Hypothesis/Objectives – This study was designed to determine the ability of canine hair follicle bulge cell‐enriched keratinocytes to construct canine living skin equivalents with interfollicular epidermis in vitro. Animals – Four healthy beagle dogs from a research colony. Methods – Bulge cell‐enriched keratinocytes showing keratin 15 immunoreactivity were isolated from canine hair follicles and cultured on dermal equivalent containing canine fibroblasts. Skin equivalents were subjected to histological, immunohistochemical, western blot and RT‐PCR analyses after 10–14 days of culture at the air–liquid interface. Results – The keratinocyte sheets showed an interfollicular epidermal structure comprising four to five living cell layers covered with a horny layer. Immunoreactivities for keratin 14 and desmoglein 3 were detected in the basal and immediate suprabasilar layers of the epidermis, while keratin 10 and desmoglein 1 occurred in more superficial layers. Claudin 1 immunoreactivity was seen in the suprabasalar layer of the constructed epidermis, and filaggrin monomers and loricrin were detected in the uppermost layer. Basal keratinocytes in the skin equivalent demonstrated immunoreactivity to antibodies against basement membrane zone molecules. Conclusions and clinical importance – A bulge stem cell‐enriched population from canine hair follicles formed interfollicular epidermis within 2 weeks in vitro, and thus represents a promising model for regenerative therapy of canine skin.     

Usefulness of cefovecin disk‐diffusion test for predicting mecA gene‐containing strains of Staphylococcus pseudintermedius and clinical efficacy of cefovecin in dogs with superficial pyoderma

Background –  Cefovecin has been widely used to treat skin infections in dogs. The relationship of the cefovecin disk‐diffusion test results to the presence of the mecA gene and the clinical efficacy of cefovecin have not been fully evaluated. Hypothesis/Objectives –  To determine the usefulness of an in vitro cefovecin disk‐diffusion test in predicting the presence of the mecA gene in Staphylococcus pseudintermedius, as well as the in vivo efficacy of cefovecin therapy in dogs with superficial pyoderma. Methods –  Twenty‐six S. pseudintermedius strains isolated from 22 dogs with pyoderma were used. In vitro disk‐diffusion test results of cefovecin were compared with agar‐dilution test results, the presence of the mecA gene, and the improvement in clinical scores of dogs with superficial pyoderma at 14 days post treatment. Results –  There was a significant linear correlation (r = ?0.83) between the diameter of the obvious zone of inhibition by disk diffusion and the minimal inhibitory concentration for cefovecin (P < 0.0001). Receiver operating characteristic analysis revealed that zone diameters between 25 and 27 mm exhibited better sensitivity (92.9%) and specificity (100.0%) for detection of strains carrying the mecA gene. The mean improvement in clinical scores in dogs carrying cefovecin‐resistant strains was significantly lower than in dogs carrying cefovecin‐susceptible strains (P < 0.01). Conclusions and clinical importance –  The cefovecin disk‐diffusion test with a cut‐off value estimated in this study was valuable for predicting mecA gene carriage in S. pseudintermedius, as well as the in vivo efficacy of cefovecin therapy in dogs with superficial pyoderma caused by S. pseudintermedius.