Heterologous expression of gassericin A,a bacteriocin produced by Lactobacillus gasseri LA39

Gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39, has a cyclic structure linking N‐ and C‐terminal amino acids. Gassericin A was expressed in Escherichia coli JM109 as a biotinylated fusion protein on the basis of the DNA sequence of mature bacteriocin. A positive clone accumulated the bacteriocin, with no activity, as a soluble fusion protein in the cytoplasm. After release of an N‐terminal tag with factor Xa protease, gassericin A was converted into an active peptide having N‐ and C‐termini. The total amount of purified bacteriocins (expressed and native) was 480 µg/L and 370 µg/L, respectively. However, the specific activity of expressed gassericin A was 15 AU/mg lower than that of native bacteriocin (2600 AU/mg). Although the actual Mr (molecular weight) of the expressed bacteriocin should be 5666, the peptide showed the same mobility (Mr 3800) in sodium dodecylsulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) as native cyclic gassericin A, suggesting that the expressed peptide retains compact folding of the molecule similar to that of native gassericin A.     

Etiology of the resinous stem canker ofChamaecyparis obtusa: Cistella japonica as the causal agent

In order to confirm the pathogenicity ofCistella japonica, inoculation experiments were conducted onto several coniferous trees. Resinous lesions similar to those of the resinous stem canker developed by the inoculation withCi. japonica only onChamaecyparis obtusa. Chamaecyparis trees were heavily affected when inoculated with the fungus in November, but slightly in May and August. Resin flows occurred when inocula ofCi. japonica were put into the holes made on stem barks and on pruning scars. Isolates ofCi. japonica varied greatly in the pathogenicity. On the lesions caused by the fungus, resinous and necrotic areas expanded in fusiform or oblong shape, and resin cysts were formed in inner barks. Beneath some necrotic inner barks, sapwood was suppressed in growth for the damages in cambial cells and stained.Ci. japonica was reisolated from the inoculated lesion tissues, though isolation frequency of the fungus became lower in some lesions two or more years after the inoculation. The results of these inoculations proved thatCi. japonica was the causal agent of the resinous stem canker ofCh. obtusa.     

Manufacturing oil palm fronds cement-bonded board cured by gaseous or supercritical carbon dioxide

This study dealt with the effects of a curing method that uses gaseous and supercritical CO₂. Its effects on the properties of oil palm fronds cement-bonded board manufactured by the conventional cold-press setting method were recorded. The effect of MgCl₂ as an accelerator of cement setting was also investigated. The hydration of cement was examined using X-ray diffractometry, thermal gravimetry, and scanning electron microscopy. The results are as follows. (1) High-performance cement-bonded boards made from oil palm fronds were successfully manufactured using the CO₂ curing method. (2) The curing method using either gaseous or supercritical CO₂ resulted in accelerated curing of cement (within several minutes). Accelerated formation of the hydration products (e.g., calcium carbonate and calcium silicate) is the main reason for the high strength of CO₂-cured boards. (3) The CO₂ curing technology does not require setting accelerators, which cause a decrease in the dimensional stability of cement-bonded board.     

Diagnostic imaging of intra-abdominal cyst in heifer using the computed tomography

A 10-month-old Japanese black heifer was diagnosed as having an intra-abdominal cyst using computed tomography (CT). Through a posterior ventral midline incision, the cyst was removed, and the heifer completely recovered after the surgery. CT scans enabled detection of the intra-abdominal cyst and measurements of the diameter of the cyst before the surgery.     

Immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation

To elucidate the relationship between steroidogenic hormones and developing adrenal glands, we investigated the immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation. Fetal adrenal glands were obtained from three horses at 217, 225 and 235 days of gestation. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17) and human placental aromatase cytochrome P450 (P450arom). Histologically, cortex and medulla cells were clearly observed in the three fetal adrenal gland tissue samples. P450scc and P450c17 were identified in cortex cells close to medulla cells and in some medulla cells in the fetal adrenal glands. P450arom was present in both cortex and medulla cells in the fetal adrenal glands. However, 3betaHSD was not found in any of the equine fetal adrenal gland tissue samples. These results suggest that equine fetal adrenal glands have the ability to synthesize androgen and estrogen, which may play an important physiological role in the development of equine fetal adrenal glands.     

Assay of chitinase and N-acetylglucosaminidase activity in forest soils with 4-methylumbelliferyl derivatives

Chitinase and N-acetylglucosaminidase activities in forest soils were examined by a highly sensitive method using 4-methylumbelliferyl (4MU) derivatives of N-acetyl-D-glucosamine oligomers as substrates. The method involves the fluorometric estimation of the 4-MU released through the activity of the soil enzymes when a soil sample and the substrate are incubated in a buffer. The activities of both enzymes decreased markedly with depth in the 4 forest soil profiles studied (i.e. Andosol, Podzol, 2 Cambisols). The activities of these soil enzymes were highly correlated with the carbon and nitrogen content of the soils.     

Equine nonneoplastic abnormal ovary in a draft mare with high serum anti-Müllerian hormone: a case study

We performed a standing hand-assisted laparoscopic ovariectomy in a draft mare that presented with high serum anti-Müllerian hormone (AMH) level and had an enlarged single cystic ovary. Histopathological examination revealed no tumor cell proliferation in the ovary, but the presence of a large ovarian cyst was confirmed. In the diagnosis of abnormal ovaries in mares, a comprehensive assessment should be performed, including the monitoring of ovarian morphology and biomarkers over time, to determine the disease prognosis and treatment plan. The case of this mare with a nonneoplastic abnormal ovary and increased serum AMH level was rare. We suggest that standing hand-assisted laparoscopic ovariectomy is useful for the removal of large ovaries in draft mares.     

Symptom development of the resinous stem canker caused by inoculation withCistella japonica ontoChamaecyparis obtusa

Cistella japonica was inoculated onto the stems of youngChamaecyparis obtusa trees, and the development of external and internal symptoms was investigated for five years. Most lesions started exuding resin from May to July during the first growing season after inoculation, while a few lesions started exuding resin during the second growing season. Resin exudation lasted for two successive years on most lesions, and during successive three, four, or five years on some lesions. Resin exuded excessively in the secondary phloem of the lesions and resin cysts frequently developed there. The lesions where cambial tissue was necrosed occupied 18% of all the lesions and the resinous areas expanded to a larger size than those where resin simply exuded in the phloem.Ci. japonica was reisolated from some inoculated lesion tissues at a high frequency. The isolation frequency of the fungus from some lesion tissues two and more years later was zero or low and that ofCryptosporiopsis abietina was high. This work was supported by a grant from the Forest Agency of the Japanese Government.