Complete Reconstitution of the Apramycin Biosynthetic Pathway Demonstrates the Unusual Incorporation of a β-d-Sugar Nucleotide in the Final Glycosylation Step.

Apramycin is a widely used aminoglycoside antibiotic with applications in veterinary medicine. It is composed of a 4-amino-4-deoxy-d-glucose moiety and the pseudodisaccharide aprosamine, which is an adduct of 2-deoxystreptamine and an unusual eight-carbon bicyclic dialdose. Despite its extensive study and relevance to medical practice, the biosynthetic pathway of this complex aminoglycoside nevertheless remains incomplete.

Biosynthetic Origin of the Octose Core and Its Mechanism of Assembly during Apramycin Biosynthesis.

Apramycin is an aminoglycoside antibiotic isolated from and that has found clinical use in veterinary medicine. The apramycin structure is notable for its atypical eight-carbon bicyclic dialdose (octose) moiety. While the apramycin biosynthetic gene cluster () has been identified and several of the encoded genes functionally characterized, how the octose core itself is assembled has remained elusive.

Mechanism of -Adenosyl-l-methionine -Methylation by Cobalamin-dependent Radical -Adenosyl-l-methionine Methylase in 1-Amino-2-methylcyclopropanecarboxylic Acid Biosynthesis.

The radical -adenosyl-l-methionine (SAM) methylase Orf29 catalyzes the -methylation of SAM in the biosynthesis of 1-amino-2-methylcyclopropanecarboxylic acid. Here, we determined that the methylation product is (4″)-4″-methyl-SAM. Furthermore, we found that the 5'-deoxyadenosyl radical generated by Orf29 abstracts the hydrogen atom from the C-4″ position of SAM to generate the radical intermediate, which reacts with methylcobalamin to give (4″)-4″-methyl-SAM.

Identification of the Early Steps in Herbicidin Biosynthesis Reveals an Atypical Mechanism of -Glycosylation.

Herbicidins are adenosine-derived nucleoside antibiotics with an unusual tricyclic core structure. Deletion of the genes responsible for formation of the tricyclic skeleton in sp. L-9-10 reveals the o importance of Her4, Her5, and Her6 in the early stages of herbicidin biosynthesis.

Experiences of Game-Based Learning and Reviewing History of the Experience Using Player's Emotions.

In this paper, we discuss whether the history of a learning experience, containing action and emotion information, is useful for review of the experience in game-based learning using virtual space. We developed a game-based story generation system that automatically generates scripts in real time by using a player's emotions and actions. The system has two functions: a game-based experiential learning environment and automatic story generation.

Characterization of the cobalamin-dependent radical S-adenosyl-l-methionine enzyme C-methyltransferase Fom3 in fosfomycin biosynthesis.

Fosfomycin is a clinically used broad-spectrum antibiotic that has the structure of an oxirane ring with a phosphonic acid substituent and a methyl substituent. In nature, fosfomycin is produced by Streptomyces spp. and Pseudomonas sp.

Biochemical and Mutational Analysis of Radical -Adenosyl-L-Methionine Adenosylhopane Synthase HpnH from Reveals that the Conserved Residue Cysteine-106 Reduces a Radical Intermediate and Determines the Stereochemistry.

Adenosylhopane is a crucial precursor of C hopanoids, which are believed to modulate the fluidity and permeability of bacterial cell membranes. Adenosylhopane is formed by a crosslinking reaction between diploptene and a 5'-deoxyadenosyl radical that is generated by the radical -adenosyl-L-methionine (SAM) enzyme HpnH. We previously showed that HpnH from A3(2) (ScHpnH) converts diploptene to (22)-adenosylhopane.

A Screening Method Using Anomaly Detection on a Smartphone for Patients With Carpal Tunnel Syndrome: Diagnostic Case-Control Study.

Background: Carpal tunnel syndrome (CTS) is a medical condition caused by compression of the median nerve in the carpal tunnel due to aging or overuse of the hand. The symptoms include numbness of the fingers and atrophy of the thenar muscle. Thenar atrophy recovers slowly postoperatively; therefore, early diagnosis and surgery are important.

-Methylation of -adenosyl-L-Methionine Occurs Prior to Cyclopropanation in the Biosynthesis of 1-Amino-2-Methylcyclopropanecarboxylic Acid (Norcoronamic Acid) in a Bacterium.

Many pharmacologically important peptides are bacterial or fungal in origin and contain nonproteinogenic amino acid (NPA) building blocks. Recently, it was reported that, in bacteria, a cyclopropane-containing NPA 1-aminocyclopropanecarboxylic acid (ACC) is produced from the L-methionine moiety of -adenosyl-L-methionine (SAM) by non-canonical ACC-forming enzymes. On the other hand, it has been suggested that a monomethylated ACC analogue, 2-methyl-ACC (MeACC), is derived from L-valine.

Effects of the Community Cats Program on Population Control, Migration and Welfare Status of Free-Roaming Cats in Tokyo, Japan.

The community cats program (CCP), which includes trap-neuter-return activities, has been promoted in Japan to reduce the population of free-roaming cats without harmful effects on their welfare. To ascertain the effects of the CCP, a two-year route census of free-roaming cats was conducted in an area with CCP and the other area without CCP in urban Tokyo, Japan. The estimated number of cats was lower in the CCP area than the non-CCP area, but there was no difference in the rate of decline in cat populations between areas.

Characterization of Radical SAM Adenosylhopane Synthase, HpnH, which Catalyzes the 5'-Deoxyadenosyl Radical Addition to Diploptene in the Biosynthesis of C Bacteriohopanepolyols.

Adenosylhopane is a crucial intermediate in the biosynthesis of bacteriohopanepolyols, which are widespread prokaryotic membrane lipids. Herein, it is demonstrated that reconstituted HpnH, a putative radical S-adenosyl-l-methionine (SAM) enzyme, commonly encoded in the hopanoid biosynthetic gene cluster, converts diploptene into adenosylhopane in the presence of SAM, flavodoxin, flavodoxin reductase, and NADPH. NMR spectra of the enzymatic reaction product were identical to those of synthetic (22R)-adenosylhopane, indicating that HpnH catalyzes stereoselective C-C formation between C29 of diploptene and C5' of 5'-deoxyadenosine.

Functional Characterization of 3-Aminobenzoic Acid Adenylation Enzyme PctU and UDP-N-Acetyl-d-Glucosamine: 3-Aminobenzoyl-ACP Glycosyltransferase PctL in Pactamycin Biosynthesis.

Pactamycin is an antibiotic produced by Streptomyces pactum with antitumor and antimalarial properties. Pactamycin has a unique aminocyclitol core that is decorated with 3-aminoacetophenone, 6-methylsaliciate, and an N,N-dimethylcarbamoyl group. Herein, we show that the adenylation enzyme PctU activates 3-aminobenzoic acid (3ABA) with adenosine triphosphate and ligates it to the holo form of the discrete acyl carrier protein PctK to yield 3ABA-PctK.

Biochemical and Structural Analysis of FomD That Catalyzes the Hydrolysis of Cytidylyl ( S)-2-Hydroxypropylphosphonate in Fosfomycin Biosynthesis.

In fosfomycin biosynthesis, the hydrolysis of cytidylyl ( S)-2-hydroxypropylphosphonate [( S)-HPP-CMP] to afford ( S)-HPP is the only uncharacterized step. Because FomD is an uncharacterized protein with a DUF402 domain that is encoded in the fosfomycin biosynthetic gene cluster, FomD was hypothesized to be responsible for this reaction. In this study, FomD was found to hydrolyze ( S)-HPP-CMP to give ( S)-HPP and CMP efficiently in the presence of Mn or Co.

C-Methylation Catalyzed by Fom3, a Cobalamin-Dependent Radical S-adenosyl-l-methionine Enzyme in Fosfomycin Biosynthesis, Proceeds with Inversion of Configuration.

Fom3, a cobalamin-dependent radical S-adenosyl-l-methionine (SAM) methyltransferase, catalyzes C-methylation at the C2 position of cytidylylated 2-hydroxyethylphosphonate (HEP-CMP) to afford cytidylylated 2-hydroxypropylphosphonate (HPP-CMP) in fosfomycin biosynthesis. In this study, the Fom3 reaction product HPP-CMP was reanalyzed by chiral ligand exchange chromatography to confirm its stereochemistry. The Fom3 methylation product was found to be ( S)-HPP-CMP only, indicating that the stereochemistry of the C-methylation catalyzed by Fom3 is ( S)-selective.

Fosfomycin Biosynthesis via Transient Cytidylylation of 2-Hydroxyethylphosphonate by the Bifunctional Fom1 Enzyme.

Fosfomycin is a wide-spectrum phosphonate antibiotic that is used clinically to treat cystitis, tympanitis, etc. Its biosynthesis starts with the formation of a carbon-phosphorus bond catalyzed by the phosphoenolpyruvate phosphomutase Fom1. We identified an additional cytidylyltransferase (CyTase) domain at the Fom1 N-terminus in addition to the phosphoenolpyruvate phosphomutase domain at the Fom1 C-terminus.

Methylcobalamin-Dependent Radical SAM C-Methyltransferase Fom3 Recognizes Cytidylyl-2-hydroxyethylphosphonate and Catalyzes the Nonstereoselective C-Methylation in Fosfomycin Biosynthesis.

A methylcobalamin (MeCbl)-dependent radical S-adenosyl-l-methionine (SAM) methyltransferase Fom3 was found to catalyze the C-methylation of cytidylyl-2-hydroxyethylphosphonate (HEP-CMP) to give cytidylyl-2-hydroxypropylphosphonate (HPP-CMP), although it was originally proposed to catalyze the C-methylation of 2-hydroxyethylphosphonate to give 2-hydroxypropylphosphonate in the biosynthesis of a unique C-P bond containing antibiotic fosfomycin in Streptomyces. Unexpectedly, the Fom3 reaction product from HEP-CMP was almost a 1:1 diastereomeric mixture of HPP-CMP, indicating that the C-methylation is not stereoselective. Presumably, only the CMP moiety of HEP-CMP is critical for substrate recognition; on the other hand, the enzyme does not fix the 2-hydroxy group of the substrate and either of the prochiral hydrogen atoms at the C2 position can be abstracted by the 5'-deoxyadenosyl radical generated from SAM to form the substrate radical intermediates, which react with MeCbl to afford the corresponding products.

Role of oxytocin in improving the welfare of farm animals - A review.

Recently, increasing attention has been paid to the welfare of farm animals, which have been evaluated using behavioral and physiological measures. However, so far, the measures have almost always been used to estimate poor welfare. In this review, firstly we focus on how oxytocin (OT) relates to positive social behavior, pleasure, and stress tolerance, and second on which management factors stimulate OT release.

The Effects of Components of Grazing System on Welfare of Fattening Pigs.

The objective of this study was to clarify the most effective component of grazing for improving welfare of fattening pigs. This study compared welfare indicators of 20 fattening pigs aged 100 to 124 days (the prior period) and 138 to 164 days (the latter period) in an indoor housing system (IS), an outdoor pasturing system (OP), a concrete floor paddock system (CF), a concrete floor paddock system with fresh grass (FG), or a soil floor paddock system (SF). The last three treatments include important components of a grazing system: extra space, grass feed, and soil floor.

Effect of Suckling Systems on Serum Oxytocin and Cortisol Concentrations and Behavior to a Novel Object in Beef Calves.

We investigated differences between effects of natural- and bucket-suckling methods on basal serum oxytocin (OT) and cortisol concentrations, and the effect of OT concentration on affiliative and investigative behavior of calves to a novel object. Ten Japanese Black calves, balanced with birth order, were allocated evenly to natural-suckling (NS) and bucket suckling (BS) groups. Blood samples were collected at the ages of 1 and 2 months (1 week after weaning) calves, and serum OT and cortisol concentrations were measured using enzyme-linked immunosorbent assay and enzymeimmunoassay tests, respectively.

Distribution of radioactive cesium and stable cesium in cattle kept on a highly contaminated area of Fukushima nuclear accident.

Radioactivity inspection of slaughtered cattle is generally conducted using a portion of the neck muscle; however, there is limited information about the distribution of radioactive cesium in cattle. In this study, therefore, we measured not only radioactive cesium but also stable cesium in various tissues of 19 cattle that had been kept in the area highly contaminated by the Fukushima nuclear accident. Skeletal muscles showed approximately 1.

Bovine growth hormone gene polymorphism affects stress response in Japanese Black cattle.

We investigate the associations between growth hormone (GH) gene polymorphism and behavioral and physiological responses to stressors and learning ability in Japanese Black cattle. Flight distance test was conducted in the first experiment. Steers with haplotype C of GH gene polymorphism avoided human approaches at a significantly greater distance than ones without haplotype C (C: 1.

Individual difference in serum oxytocin concentrations of calves and the correlation with those in dams.

We examined individual differences in serum oxytocin concentrations (OT) of calves, and assessed whether these differences were correlated with their dams' milk and serum OT. Eight Holstein (H), nine Japanese Shorthorn (JS), and six Japanese Black (JB) calves were examined. Blood was collected three times during the first month in H calves, while their dams' blood was collected three times prior to parturition.

Distribution of radioactive cesium in edible parts of cattle.

After the disastrous incident of the Fukushima Daiichi Nuclear Power Station, various agricultural, livestock and fishery products have been inspected for radioactive contamination with cesium in Japan. In this study, radioactive cesium was measured in various edible parts of cattle to verify the current inspection method for cattle, in which the neck tissues are generally used as samples. Radioactive cesium concentration in the short plate, diaphragm, liver, lung, omasum, abomasum and small intestine were lower and sirloin, tenderloin, top round meat and tongue were higher than that in the neck.

Assessment of the effect of presenting a companion's face picture on social isolation stress using saliva sampling in cows.

The effect of presenting a companion's face picture on social isolation stress was assessed by saliva sampling in cows. Three Japanese Black test cows and another companion cow were used. At 09.

Effects of Na-butyrate supplementation in milk formula on plasma concentrations of GH and insulin, and on rumen papilla development in calves.

Although the growth-promoting action of sodium-butyrate (Na-butyrate) used as a feed additive has been observed in calves and pigs, the precise mechanisms involved remain to be clarified. In this study, pre-weaning calves were given milk formula (MF) supplemented with butyrate for 6 weeks to investigate its effects on postprandial changes in the plasma concentrations of metabolic hormones, and, simultaneously, on growth performance, the weight of the digestive organs and rumen papilla development. Ingestion of MF increased (P<0.