Molecular cloning, tissue expression, and subcellular localization of porcine peptidoglycan recognition proteins 3 and 4

Peptidoglycan recognition proteins (PGRPs) are innate immune molecules that are present in most invertebrates and vertebrates. Mammals have four PGRPs, PGLYRP1-4. In the present study, we cloned the cDNAs encoding porcine PGLYRP3 and 4 from the esophagus of adult swine. The length of the complete open reading frames of porcine PGLYRP3 and 4 are identical and contain 1125bp encoding 374 amino acid residues. The amino acid sequences of these two proteins were more similar to their human orthologs (78.9% [PGLYRP3] and 73.9% [PGLYRP4]) than to their mouse orthologs (71.3% [PGLYRP3] and 67.9% [PGLYRP4]). Expression analysis revealed that both PGLYRP3 and 4 were more strongly expressed in digestive tract, especially the esophagus, than in immune organs such as spleen or mesenteric lymph nodes in both newborn and adult swine. To analyze the subcellular distribution of porcine PGLYRP1-4, we constructed transfectant cell lines. Western blot and flow cytometric analyses revealed that porcine PGLYRP3 and 4 are not only secreted, but also expressed on the cell surface, unlike PGLYRP1 and 2. These results should help contribute to the understanding of PGLYRP3- and 4-mediated immune responses via their recognition of intestinal microorganisms in newborn and adult swine.     

Chemical characterization of milk oligosaccharides of the island flying fox (Pteropus hypomelanus) (Chiroptera: Pteropodidae)

Although a considerable amount of information has accumulated about oligosaccharides in the milk and colostrum of representatives of various mammalian orders, nothing is so far known concerning these sugars in the milk of any bat species (order Chiroptera). In this study, we determined that the following oligosaccharides occur in milk of the island flying fox, Pteropus hypomelanus (Chiroptera: Pteropidae): Gal(α1–3)Gal(β1–4)Glc (isoglobotriose), Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (lacto‐N‐neotetraose), Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (lacto‐N‐neohexaose) and Neu5Gc(α2–3)Gal(β1–4)Glc (3′‐NGc‐SL). However, lactose was found to be the dominant saccharide in this milk, as in most eutherian mammals. The biologic importance of oligosaccharides in Chiropteran milks warrants further study.     

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.     

Relative importance of protons and solution calcium concentration in phosphate rock dissolution by organic acids

A series of experiments was conducted to quantify the relative contribution of protons and other mechanisms to the dissolution of phosphate rocks (PRs) from six countries in solutions of low-molecular-weight-aliphatic organic acids. The amounts of P and Ca released after 3 d of incubation at 28°C were determined in all the experiments. In the first experiment the solubility of the PRs < 500 μm particle size) in 100 M (25 mL g^-1 PR) oxalic, tartaric, and citric acids was compared with that in three mineral acids and four chelating compounds. There were no differences in the amount of P released by the mineral acids, but the organic acids released more P than could be accounted for by protonation. The chelating compounds were the least effective. In the second experiment, 1 g each of Sri Lanka and Togo PRs was incubated with 2.5 mmol of the organic acids using three acid concentration (mM) : acid volume ratios (250:10, 100:25, and 25:100). The amount of P dissolved from the PRs and the amount of acid remaining in the filtrate were about the same for all three treatments. A mixture of the organic acids and the respective Na-salts was prepared at six different acid: salt ratios in the third experiment. Total acid concentration in all the solutions was 100 mM. Higher proportion of the Na-salt in the mixture decreased PR dissolution, while the amount of the acid remaining in the filtrate increased. The Ca concentration in the filtrate showed an opposite trend. The results indicated that PR dissolution by the organic acids was initially dependent on protonation, which accounted for only 13–38% of the amount of P dissolved. Most of the amount of P dissolved could therefore be attributed to other mechanisms. Removal of dissolved Ca from the solution appears to be the major factor controlling the subsequent dissolution of the PRs. The ability of tartaric and oxalic acids to dissolve effectively the PRs was attributed to the formation of an insoluble calcium compound which was precipitated from the solution. For the use of organic acids as a potential amendment for improving the P availability of PRs, the Ca binding power of the organic acids should also be considered.     

Historical aspects of soil classification in Japan

Historical perspective of soil classification in Japan from Max Fesca's soil classification in 1882 to the “Unified Soil Classification System of Japan (2002)” was outlined, aiming at reviewing the progress in soil classification. The evolution can be divided into the following five aspects: 1) Max Fesca's soil texture survey and soil classification from the agro-geological point of view under the influence of the German school; 2) Introduction of the concept of pedology into the classification under the influence of the Russian school led by Dokchaev; 3) Brief history of the classification of Andosols which has exerted a considerable influence on soil classification worldwide; 4) Soil classifications developed through the implementation of national soil survey projects to independently evaluate land suitability for the cultivation of paddy rice, upland crops, and for forest establishment; 5) Attempts to develop a comprehensive soil classification system in order to unify soil classification systems for the above-mentioned land uses from 1963 to the present.     

The membrane lipid bilayer as a regulated barrier to cope with detrimental ionic conditions: making new tolerant plant lines with altered membrane lipid bilayer

Several detrimental ionic conditions can occur in crop fields: soil acidity, salinity, heavy metal toxicity, and/or nutrient deficiency. Crop plants tolerant to these detrimental ionic conditions have excellent strategies that are related to external and/or internal mechanisms. Recently, we proposed a new concept of aluminum (Al) tolerance in plants; specifically, a plasma membrane (PM) lipid bilayer mechanism. This mechanism is defined as the retardation of Al permeation through the PM lipid bilayer based on the specific composition of the lipid molecules in the PM. The molecular basis of a less negatively charged PM lipid bilayer is smaller proportions of phospholipids and greater proportions of galactolipids and sterols. This leads to reduced adsorbability of detrimental ions onto the PM lipid bilayer, resulting in less permeabilization. Phenolics and carotenoids have two moieties; a hydrophilic moiety and a hydrophobic moiety. The hydrophobic moieties of these compounds can occlude the permeabilized spaces in the PM lipid bilayer, thereby reinforcing it. Another strategy to retard the permeability of the PM to detrimental ions is to decrease the proportion of stigmasterol, which has been shown to have no ability to reduce water permeability. The beneficial or harmful effects of various organic materials (organic fertilizers, soil organic matter, agrochemicals, or organic pollutants) on the productivity or quality of crop plants in relation to changes in the PM lipid bilayer are discussed. Modulation of the PM lipid bilayer is a promising strategy to produce new crop lines tolerant to detrimental ionic conditions.     

Electrocardiographic evaluation of the degree of sedation and the isolated use of methadone in healthy dogs

ObjectiveThe present study aimed to investigate the influence of methadone on cardiorespiratory parameters, electrocardiogram and clinical sedation in dogs. Further possible side effects are reported.Study designProspective experimental cross-over study.DogsEight, 1–4-year-old, various breeds of dogs of both genders weighing 9–36 kg.MethodsEach dog was treated three times: methadone 0.3 mg kg^?1 (M0.3), 0.5 mg kg^?1 (M0.5) and 1.0 mg kg^?1 (M1.0) intramuscularly. Respiratory rate, heart rate and arterial blood pressure were recorded as well as electrocardiographic evaluation of lead II. Clinical sedation in each treatment received a score (0–3) after drug administration and at 30 minute intervals until scores and measurements returned to baseline values.ResultsA significant decrease in heart rate was seen with each dose of methadone and bradycardia (HR<60 bpm) was noted in a few dogs at each dose. A clinically significant arrhythmia occurred in one dog at 1 mg kg^?1 that required reversal with butorphanol. There was no significant difference in SAP, MAP and DAP between treatments. Some side effects such as salivation, defecation, vocalization and panting, after administration of methadone were observed. There were no differences in mean values of heart rate, P-wave and QRS complex duration and QT interval between treatments.Conclusion and clinical relevanceMethadone administration was associated with panting and a decrease in heart rate at all doses tested in this study. The cardiac rhythm should be monitored carefully in dogs when methadone is administered on its own, especially at higher doses.     

Tumescent local anesthesia with ropivacaine in different concentrations in bitches undergoing mastectomy: plasma concentration and post‐operative analgesia

ObjectiveTo compare two concentrations of ropivacaine administered for tumescent local anesthesia (TLA) in dogs undergoing mastectomy.Study designProspective randomized clinical study.AnimalsSeventeen bitches of various breeds, aged 12 ± 2 years and weighing 10 ± 6.5 kg requiring total unilateral or bilateral mastectomy.MethodsDogs were premedicated with acepromazine (0.04 mg kg^?1) and morphine (0.4 mg kg^?1) intramuscularly. Anesthesia was induced with propofol (2.5 mg kg^?1) and midazolam (0.2 mg kg^?1) intravenously, followed by intubation and maintenance with isoflurane and TLA. Dogs were randomly allocated to receive TLA either with 0.1% ropivacaine (group G1) or with 0.05% ropivacaine (group G05). TLA was performed by insertion of a multihole needle under the skin and infusion of ropivacaine and lactated Ringer’s solution at a fixed volume of 15 mL kg^?1. Ropivacaine concentrations in arterial blood were measured by high-performance liquid chromatography. Post-operative pain was assessed using two scales (University of Melbourne pain scale and a modified composite measure pain scale) and von Frey filaments, 4 hours after TLA and at 1 hour intervals until sensitivity was regained. A score above 30% of the maximum possible score was considered a positive indicator of pain.ResultsPeak plasma concentrations of ropivacaine were measured 240 minutes after TLA in G1. Low concentrations were measured in G05 for 60 minutes, with subsequent increase. Analgesic rescue and return of sensitivity occurred at 7 ± 2.3 and 7 ± 1.9 hours (mean ± SD) after TLA for G1 and G05, respectively.Conclusions and clinical relevanceTumescent local anesthesia with ropivacaine provided satisfactory post-operative analgesia that lasted for several hours, with no difference in duration between the concentrations. No serious side effects were attributed to TLA. Results indicated that 0.05% ropivacaine provided adequate analgesia for mastectomy, however, more studies are required to support this conclusion.     

Neutral and acidic milk oligosaccharides of the striped skunk (Mephitidae: Mephitis mephitis)

The biological significance of the tremendous variation in proportions of oligosaccharides and lactose among mammalian milks is poorly understood. We investigated milk oligosaccharides of the striped skunk (Mephitis mephitis) and compared these results to other species of the clade Mustelida. Individual oligosaccharides were identified by proton nuclear magnetic resonance spectroscopy. In the striped skunk, six oligosaccharides were identified: isoglobotriose, 2′‐fucosyllactose, A‐tetrasaccharide, Galili pentasaccharide, 3′‐sialyllactose and monosialyl monogalactosyl lacto‐N‐neohexaose. Four of these have been found in related Mustelida and the other two in more distantly related carnivorans. The neutral and acidic oligosaccharides derive from three core structures: lactose (Gal(β1–4)Glc), lacto‐N‐neotetraose (Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc) and lacto‐N‐neohexaose (Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc).     

Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH)

Adhesion tests are complex, time‐consuming and expensive, while the most important criterion for a probiotic lactobacilli is the ability to adhere to the human intestine. Thirty lactobacilli isolates from human intestinal tissues were measured for cell surface glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) activity using a microtiter plate screening method. GAPDH activities were detected in 21 out of 30 samples from 12 h cultures and in all samples from 18 h cultures. This suggests GAPDH is universally expressed on the bacterial cell surfaces from many lactobacilli. A statistically significant positive correlation was shown between GAPDH activity and adhesion using the BIACORE adhesion assay (P < 0.01). The new screening method using GAPDH enzymatic activity without an adhesion test may be possible due to the significant positive correlation of GAPDH activity with adhesion of lactobacilli derived from the human intestine.