Immobilization of glucosyltransferase fromAureobasidium

Summary Glucosyltransferase fromAureobasidium, which produces panose and isomaltose from maltose, was immobilized by alginate gel or DEAE-cellulose at high efficiency (71 and 41% respectively). Alkylamine porous silica was less efficient as a support. The enzymatic profiles of immobilized enzymes were almost identical to the native one except that their stabilities to extreme pH, metal ions and inhibitors were improved. Both immobilization procedures successfully produced high amounts of panose, 125 mg ml^–1 (alginate gel) or 141 mg ml^–1 (DEAE-cellulose), from 300 mg ml^–1 of maltose.     

Long-term continuous reaction of immobilized β-fructofuranosidase

Summary -Fructofuranosidase was immobilized by alginate gel at high efficiency (92 %). The extreme long-term continuous reaction (half-life, 275 days) was achieved by the immobilized enzyme using sucrose at high concentration (500 mg ml^–1) to produce fructo-olicosaccharides, such as 1-kestose (Fru21Fru21aGlc) and nystose (Fru21Fru21Fru21aGlc).     

Observation of the chemical structure of fructooligosaccharide produced by an enzyme fromAureobasidium sp. ATCC 20524

The chemical structure of the oligosaccharide produced from sucrose by an enzyme extracted fromAureobasidium sp. ATCC 20524 was observed. The GC-MS analysis by methylation indicated that this oligosaccharide is composed of 2-linked, 1,2-linked fructose and 1-linked glucose. The [¹³C]-NMR spectrum indicated that the 1,2-linked glycosidic linkages of fructose of this oligosaccharide are, and the 1-linked glycosidic linkages of fructose are . This investigation suggested that this oligosaccharide is isokestose.     

Influence of Pneumococcal Conjugate Vaccine on Acute Otitis Media with Severe Middle Ear Inflammation: A Retrospective Multicenter Study

The Japanese guidelines for acute otitis media in children recommend classifying acute otitis media by age, manifestations and local findings, and also recommend myringotomy for moderate-grade cases with severe local findings, severe-grade cases, and treatment-resistant cases. The heptavalent pneumococcal conjugate vaccine was released in Japan in February 2010. In Hiroshima City, public funding allowing free inoculation with this vaccine was initiated from January 2011, and the number of vaccinated individuals has since increased dramatically. This study investigated changes in the number of myringotomies performed to treat acute otitis media during the 5-year period from January 2008 to December 2012 at two hospitals and five clinics in the Asa Area of Hiroshima City, Japan. A total of 3,165 myringotomies for acute otitis media were performed. The rate of procedures per child-year performed in <5-year-old children decreased by 29.1% in 2011 and by 25.2% in 2012 compared to the mean rate performed in the 3 years prior to the introduction of public funding. A total of 895 myringotomies were performed for 1-year-old infants. The rate of myringotomies per child-year performed for acute otitis media in 1-year-old infants decreased significantly in the 2 years after the introduction of public funding for heptavalent pneumococcal conjugate vaccine compared to all years before introduction (p<0.000001). Our results suggest a benefit of heptavalent pneumococcal conjugate vaccine for acute otitis media in reducing the financial burden of myringotomy. In addition, this vaccine may help prevent acute otitis media with severe middle ear inflammation in 1-year-old infants.     

Spatial pattern changes of grazing cattle population I

A cattle population presents three spatial patterns according to their behaviour. These patterns are as follows: the random spatial pattern which occurs when the cattle move during grazing (feeding), the aggregated spatial pattern which occurs when they rest, and the regular or quasi-regular spatial pattern which occurs when they separate into two or more subgroups. In this study, these three states of behaviour were represented by a mathematical model, and a method to test the pattern statistically was derived.     

Plasma phosphatidylcholine hydroperoxide as a new marker of oxidative stress in alcoholic patients

Quantitative analysis of plasma phosphatidylcholine hydroperoxide (PCOOH) is an important step in evaluating the biochemical processes leading to oxidative injury. However, secondary products of lipid peroxidation are now used as indices. One hundred nine alcoholic patients, aged 22-81 years (mean +/- SEM, 52.0 +/- 1.3 years), and 21 healthy volunteers, aged 41-79 years (51.2 +/- 2.2 years), participated in this study. Plasma PCOOH was measured by HPLC with chemiluminescence detection. Plasma PCOOH concentration was significantly higher in alcoholic patients (46.1 +/- 4.1 pmol/ml) than in controls (15.6 +/- 1.8 pmol/ml). It was significantly higher in patients with blood alcohol (88.0 +/- 10.5 pmol/ml) than in those without alcohol (32.6 +/- 3.1 pmol/ml). The patients with high levels of aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transpeptidase (gamma-GTP), and triglyceride (TG) showed significantly higher PCOOH concentrations than did patients with normal levels. The PCOOH level was positively correlated with levels of gamma-GTP, HDL, blood alcohol concentration, and TG. Plasma PCOOH levels in 29 alcoholic patients after a 6 week abstinence were decreased significantly (22.8 +/- 11.1 pmol/ml), which was associated with improvement on liver function tests. This is the first measurement of plasma PCOOH in alcoholic patients. These results suggest the involvement of lipid peroxidation in alcohol-induced liver damage and confirm that the PCOOH plasma concentration is a new marker of alcohol consumption as well as oxidative stress in alcoholic patients.     

Genetic diversity of Gambierdiscus spp. (Gonyaulacales, Dinophyceae) in Japanese coastal areas

The genetic diversity of the ciguatera fish poisoning-related dinoflagellate distributed in Japanese coastal areas was investigated. The entire sequence of the 5.8S rRNA gene and two internal transcribed (ITS) regions were determined, which included putative pseudogenes, from 19 strains of dinoflagellates assigned to the genus Gambierdiscus Adachi et Fukuyo collected from Japanese subtropical and temperate coastal areas. The sequences obtained from the 19 strains were divided into two types based on sequence similarity. Here we designate the two types as type 1 and type 2. For the relationship between the genotypes and origins of the strains used, the strains collected from subtropical areas possessed the type 1 sequence; whereas those from temperate areas possessed the type 2. This observation led us to question former reputations that Gambierdiscus cells observed in Japanese temperate areas are immigrants from Japanese subtropical areas. Subsequently, we sequenced a part of the 18S rRNA gene from two strains from subtropical areas and two from temperate areas. Unfortunately, phylogenetic analysis including the sequences obtained from various gonyaulacales dinoflagellates failed to determine the species phylogenetically closely related to and possible origin(s) of the Gambierdiscus sp. from the Japanese coastal areas.     

Enhanced stretch reflex excitability in the soleus muscle during passive standing posture in humans

The purpose of this study was to test whether the spinal reflex excitability of the soleus muscle is modulated as posture changes from a supine to a passive upright position. Eight healthy subjects (29.6 ± 5.4 yrs) participated in this study. Stretch and H-reflex responses were elicited while the subjects maintained passive standing (ST) and supine (SP) postures. The passive standing posture was accomplished by using a gait orthosis to which a custom-made device was mounted to elicit stretch reflex in the soleus muscle. This orthosis makes it possible to elicit stretch and H-reflexes without background muscle activity in the soleus muscle. The results revealed that the H-reflex amplitude in the ST was smaller than that in the SP condition, which is in good agreement with previous reports. On the other hand, the stretch reflex was significantly larger in the ST than in the SP condition. Since the experimental conditions of both the stretch and H-reflex measurements were exactly the same, the results were attributed to differences in the underlying neural mechanisms of the two reflex systems: different sensitivity of the presynaptic inhibition onto the spinal motoneuron pool and/or a change in the muscle spindle sensitivity.     

On freeze-thaw sequence of vital organ of assuming the cryoablation for malignant lung tumors by using cryoprobe as heat source

Regarding cryoablation for the malignant lung tumors, multiple trials of the freeze-thaw process have been made, and we considered it necessary to view and analyze the freeze-thaw process as a freeze-thaw sequence. We caused the sequence in a porcine lung in vivo by using an acicular, cylindrical stainless-steel probe as the heat source for the freeze-thaw sequence and cooling to -150 °C with super high-pressure argon gas by causing the Joule-Thomson effect phenomenon at the tip of the probe. In this experiment, we examined the sequence by measuring the temperature and using the isothermal curve and the freezing function. As a result, it was demonstrated that the freezing characteristics considerably differed in the first sequence and the second sequence from those of non-aerated organs such as liver and kidney. In our experiments on porcine lung, thermal properties were considered to change as the bleeding caused by the first thawing infiltrated in the lung parenchyma, and it was confirmed that the frozen area in the second cycle was dramatically enlarged as compared with the first cycle (when a similar sequence is continuously repeated, we say it as cycle). This paper provides these details.     

Synthesis and Antimicrobial Activity of Optically Active trans-Cycloheximide Isomers

Optically active tiraras-cycloheximide isomers such as cycloheximide [(2S,4S,6R,αR)-form (1)], naramycin B[(25,4S,6RαR)-form(4)], and new stereoisomers (2S,4S,6S,αS)-form (8) and (2S,4S,6R,αS)-from (9) were synthesized by an aldol condensation of trans-2,4-dimethyl-l-cyclohexanone (5b), with 4-(2-oxoethyl)-2,6-piperidinedione(6). The antimicrobial activity of trans- cycloheximide isomers (1, 4, 8, and 9) was examined against S. cerevisiae and P. oryzae. The stereoisomers 1 and 4 exhibited marked antimicrobial activity against both microorganisms as compared with their C- α-epimers 8 and 9.