Using the indicator amino acid oxidation technique to study threonine requirements in horses receiving a predominantly forage diet

Threonine has been reported to be the second limiting amino acid in typical equine diets, but its actual requirement has not been determined in horses. To evaluate amino acid metabolism and requirements, the indicator amino acid oxidation (IAAO) method has been successfully used in other species. The objective of this research was to estimate threonine requirements in mature horses fed timothy hay and concentrate in 4:1 ratio using the IAAO method. Six Thoroughbred mares (579.9 ± 46.7 kg) received each of 6 levels of threonine intake, 41, 51, 61, 70, 80 and 89 mg/kg BW/day, in a randomly determined order. Each study period was 7‐day long, and on day 6, blood samples were collected before and 90 min after feeding to measure amino acid concentrations using HPLC. On day 7, horses underwent IAAO procedures, which included a 2‐hr primed, constant intravenous infusion of [¹³C]sodium bicarbonate to measure total CO₂ production and a 4‐hr primed, constant oral administration of [1‐¹³C]phenylalanine to estimate phenylalanine oxidation to CO₂. Blood and breath samples were collected to measure blood [¹³C]phenylalanine, using GC‐MS analysis and breath ¹³CO₂ enrichment, using an infrared isotope analyser. Increasing threonine intake levels did not affect plasma phenylalanine oxidation by the ANOVA test (p > 0.05) but resulted in a linear decrease in phenylalanine oxidation (p = 0.04) without a breakpoint by the orthogonal linear contrast. This study is the first attempt to evaluate threonine requirements in horses by the IAAO method; however, threonine requirements are still unknown in mature horses at this time.     

Contrast enhanced ultrasonography of kidney in conscious and anesthetized beagle dogs

Contrast enhanced ultrasound (CEUS) is useful to evaluate tissue perfusion in the kidney. In veterinary medicine, sedation or anesthesia may be required in uncooperative or panting patients. The aim of this study was to evaluate and compare the normal kidney perfusion patterns in conscious and anesthetized dogs using CEUS. Eight healthy beagles were used in this study. Scanning was performed in conscious dogs using manual restraint (conscious group), or under general anesthesia using tiletamine-zolazepam and medetomidine (TZM group) or medetomidine (M group). The contrast agent (Sonovue^®) was administered as an IV bolus. The peak intensity (PI), time to peak enhancement from injection (TTP₀) and the time to peak enhancement from the initial rise (TTP_up), upslope, downslope and area under the curve (AUC) were analyzed. Compared to the cortical values in the conscious group, TTP₀ was significantly delayed in the TZM group, and upslope, TTP₀ and TTP_up were significantly different in the M group. The AUCs in the TZM and M groups were not different from those in the conscious group. The upslope of renal medullary perfusion was significantly decreased in the TZM and M groups. TTP₀ and TTP_up were also significantly delayed in these groups. The AUC of the medulla was significantly decreased in the M group. Therefore, TZM is useful as an anesthetic protocol when performing CEUS, and the obtained data may serve as reference values in the evaluation of renal perfusion using CEUS in dogs under anesthesia.     

Evaluation of left ventricular function with cardiac magnetic resonance imaging and echocardiography after administration of dobutamine and esmolol in healthy beagle dogs

Unlike echocardiography, cardiac magnetic resonance imaging (cardiac MRI) results in a near-exact assessment of cardiac structures and function. However, most veterinary studies have focused on dogs with normal cardiac function. We hypothesized that there would be significant differences in cardiac measurements between cardiac MRI and echocardiography when left ventricular (LV) function was abnormal. This study was undertaken to compare measurements of LV function produced by cardiac MRI and echocardiography in dogs whose LV function was altered by pharmacological agents. This study was conducted with six healthy beagle dogs. We increased left ventricular contractility by administration of dobutamine; we decreased cardiac contractility with esmolol. Stroke volume measurements were made by using both cardiac MRI and echocardiography under seven different conditions with general anesthesia: control, three doses of esmolol (100, 200, and 500 µg/kg/min), and three doses of dobutamine (10, 20, and 50 µg/kg/min). Experiments involving each condition were conducted at least 1 week apart. When LV contractility was normal, ejection fraction (EF) and stroke volume (SV), as measured by echocardiography and cardiac MRI, were not significantly different. However, when contractility was changed by pharmacological agents, EF and SV were overestimated by echocardiography, compared to MRI. Evaluation of cardiac function in patients treated with pharmacological agents should be conducted carefully because EF and SV measured by echocardiography can be overestimated, compared with EF and SV obtained by cardiac MRI.     

Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.     

Effects of electroacupuncture on intraocular pressure and hemodynamic parameters in isoflurane anesthetized dogs

The effects of electroacupuncture (EA) on intraocular pressure (IOP) and hemodynamic parameters were evaluated in isoflurane anesthetized 10 (5 males, 5 females) normal mongrel dogs (8.1-9.8 kg, 6-8 years old). After determination of baseline IOP and hemodynamic parameters (cardiac index, systolic arterial pressure, diastolic arterial pressure, heart rate and systemic vascular resistance index), EA was applied at 3 acupoints (LI-4, LIV-3 and GB-37) for 20 min. After the EA treatment, IOP was significantly decreased in the both eyes (p<0.05). However, there were not significant differences in hemodynamic parameters between those of before and after EA treatment. From these results, the EA treatment at LI-4, LIV-3 and GB-37 would be considered one of the valuable methods for the IOP treatment in dogs.     

Growth Responses and Differences in Gene Expression depending on Cultivation Temperature between Alternative type Wheat Varieties

Journal of Crop Science and Biotechnology - Two domestic wheat varieties were grown in the growth chamber set at 17°C, 20°C, 23°C and 26°C on average after the emergence of...     

Hydroxycinnamoylmalic acids and their methyl esters from pear (Pyrus pyrifolia Nakai) fruit peel

Two novel caffeoylmalic acid methyl esters, 2-O-(trans-caffeoyl)malic acid 1-methyl ester (6) and 2-O-(trans-caffeoyl)malic acid 4-methyl ester (7), were isolated from pear (Pyrus pyrifolia Nakai cv. Chuhwangbae) fruit peels. In addition, 5 known hydroxycinnamoylmalic acids and their methyl esters were identified: 2-O-(trans-coumaroyl)malic acid (1), 2-O-(cis-coumaroyl)malic acid (2), 2-O-(cis-coumaroyl)malic acid 1-methyl ester (3), 2-O-(trans-coumaroyl)malic acid 1-methyl ester (4), and 2-O-(trans-caffeoyl)malic acid (phaselic acid, 5). The chemical structures of these compounds were determined by spectroscopic data from ESI MS and NMR. Of all the isolated compounds, five hydroxycinnamoylmalic acids and their methyl esters (2-4, 6, 7) were identified in the pear for the first time.     

Electron spin resonance and luminescence spectroscopic observation and kinetic study of chemical and physical singlet oxygen quenching by resveratrol in methanol

Electron spin resonance (ESR) spectroscopy and near-infrared (NIR) fluorescence spectroscopy were performed to observe singlet oxygen quenching by resveratrol. Resveratrol greatly decreased the 2,2,6,6-Tetramethyl-4-piperidone-N-oxyl radical signal as determined by ESR spectroscopy. Resveratrol also efficiently decreased luminescence emission at 1268 nm as studied with a NIR spectrofluorometer, showing positive evidence of singlet oxygen quenching by resveratrol. The total singlet oxygen quenching rate constant (kr+kq) of resveratrol in methanol was determined to be 2.55×10(7) M(-1) s(-1). The singlet oxygen chemical quenching rate constant (kr) of resveratrol was calculated by measuring its reaction rate with singlet oxygen relative to that of α-terpinene in the same solution under light illumination. The kr value of resveratrol was 1.15×10(6) M(-1) s(-1). The percent partition of chemical quenching over total singlet oxygen quenching (kr×100)/(kr+kq) for resveratrol was 5.11%. The results showed that resveratrol quenches singlet oxygen almost exclusively through the mechanism of physical quenching. Resveratrol showed a protective activity similar to that of BHA on the methylene blue sensitized photooxidation of α-terpinene. This unambiguously explains the mechanism of how resveratrol protects tissues and cells in biological systems or important nutrients in food systems against their photosensitized oxidations.     

The mTOR-regulated phosphoproteome reveals a mechanism of mTORC1-mediated inhibition of growth factor signaling

The mammalian target of rapamycin (mTOR) protein kinase is a master growth promoter that nucleates two complexes, mTORC1 and mTORC2. Despite the diverse processes controlled by mTOR, few substrates are known. We defined the mTOR-regulated phosphoproteome by quantitative mass spectrometry and characterized the primary sequence motif specificity of mTOR using positional scanning peptide libraries. We found that the phosphorylation response to insulin is largely mTOR dependent and that mTOR exhibits a unique preference for proline, hydrophobic, and aromatic residues at the +1 position. The adaptor protein Grb10 was identified as an mTORC1 substrate that mediates the inhibition of phosphoinositide 3-kinase typical of cells lacking tuberous sclerosis complex 2 (TSC2), a tumor suppressor and negative regulator of mTORC1. Our work clarifies how mTORC1 inhibits growth factor signaling and opens new areas of investigation in mTOR biology.     

Soil texture affects the conversion factor of electrical conductivity from 1:5 soil-water to saturated paste extracts

Electrical conductivity(EC) of soil-water extracts is commonly used to assess soil salinity. However, its conversion to the EC of saturated soil paste extracts(ECe), the standard measure of soil salinity, is currently required for practical applications. Although many regression models can be used to obtain ECe from the EC of soil-water extracts, the application of a site-specific model to different sites is not straightforward due to confounding soil factors such as soil texture. This study was...