Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Extrapulmonary granulomatous lesions in canine paragonimiasis

Extrapulmonary granulomatous lesions associated with a naturally acquired Paragonimus kellicotti infection were observed in a dog with severe pulmonary paragonimiasis. Lung lesions were similar to those previously described. Extrapulmonary granulomatous lesions were found in the liver, mediastinal lymph nodes, spermatic cord and the tunica vaginalis. Extravascular deposition of eggs in the spermatic cord and tunica vaginalis caused a perilymphatic granulomatous inflammation and partially occlusive lymphangitis with dilatation of the external lymphatic vessel of the right spermatic cord.     

Impact of Buckwheat Flavonoids on In Vitro Starch Digestibility and Noodle‐Making Properties

The effects of various buckwheat materials (buckwheat flour [BF], dietary fiber extract [DE], flavonoids extract [FE], and rutin‐enhanced flavonoids extract [REFE]) on starch digestibility and noodle‐making properties were evaluated. When FE and REFE were incorporated into noodles, the amount of rapidly digestible starch and the predicted glycemic index (pGI) were reduced. However, BF and DE did not significantly decrease the pGI value of noodles. When assessing noodle properties, hardness was increased with increasing content of buckwheat materials, whereas other texture parameters were not significantly affected by buckwheat addition. All noodles were similar in regard to water absorption and swelling index, but cooking loss was slightly increased in FE and REFE noodles. FE and REFE demonstrated higher flavonoid stability during noodle making and, additionally, were more effective at reducing starch digestibility than BF and DE. REFE, specifically, does not generate quercetin (the cause of a bitter taste), and, therefore, REFE was effective in suppressing the hydrolysis of starch in the noodles, lowering the pGI.     

Serum adipokine concentrations in dogs with diabetes mellitus: a pilot study

This study was conducted to determine whether serum adipokine concentrations differed between healthy dogs and dogs with diabetes mellitus (DM). To accomplish this, 19 dogs with newly diagnosed DM were compared to 20 otherwise healthy dogs. The serum concentrations of visfatin, leptin, IL-1β, IL-6, IL-18, and TNF-α were significantly higher in diabetic dogs than in healthy dogs, whereas the serum adiponectin concentrations were lower in diabetic dogs. However, there were no significant differences in the IL-10 and resistin levels between groups. The serum leptin concentrations in diabetic dogs with and without concurrent disorders differed significantly. Treatment with insulin induced a significant decrease in IL-6 in diabetic dogs without concurrent disorders. These results show that the clinical diabetic state of dogs could modulate the circulating visfatin and adiponectin concentrations directly, while upregulation of leptin was probably a result of concurrent disorders rather than an effect of persistent hyperglycemia as a result of DM.     

Enhancing resistance in Pinus radiata seedlings to terminal crook (Colletotrichum acutatum) using methyl jasmonate and ultraviolet‐C radiation

Colletotrichum acutatum is a fungal pathogen that causes terminal crook disease in radiata pine (Pinus radiata) seedlings in New Zealand forest nurseries. Symptoms of infection include malformation or death of the growing tip and a stiffening and thickening of the stem. Although the disease can be managed effectively using fungicides, the New Zealand forest industry is interested in alternative control options such as induced resistance. The aim of this study was to evaluate spray application of chitosan (1.4 g/l) or 2.25 mm methyl jasmonate (MeJA) and irradiation with UV‐C (2.16 kJ/m²) for their potential to induce resistance to terminal crook. The treatments were applied to 4‐month‐old seedlings at 1 week before pathogen inoculation. By the end of the experimental period (42 days after inoculation), there was 80% disease incidence in the controls, with 48% of seedlings exhibiting severe terminal crook symptoms. The most effective treatment (p < 0.05) was MeJA with 16% disease incidence and none with severe symptoms. UV‐C also significantly (p < 0.05) reduced infection with 52% incidence and 20% of seedlings exhibiting severe symptoms. Chitosan did not reduce disease incidence (72%) compared with the control (80%) but did significantly reduce (p < 0.05) disease severity with 28% exhibiting severe symptoms. MeJA was the only treatment that significantly (p < 0.05) reduced the detrimental effects of infection on seedling apical growth and stem diameter. To our knowledge, this is the first report demonstrating the potential for MeJA and UV‐C to control terminal crook in radiata pine.     

Levan fructotransferase from Arthrobacter oxydans J17-21 catalyzes the formation of the di-D-fructose dianhydride IV from levan

A new levan fructotransferase (LFTase) isolated from Arthrobacter oxydans J17-21 was characterized for the production of difructose dianhydride IV (DFA IV). LFTase was purified to apparent homogeneity by Q-Sepharose ion exchange chromatography, Mono-Q HR 5/5 column chromatography, and gel permeation chromatography. The enzyme had an apparent molecular mass of 54000 Da. The optimum pH for the enzyme-catalyzed reaction was pH 6.5, and the optimum temperature was observed at 45 degrees C. The LFTase was activated by the presence of CaCl(2) and EDTA-2Na but inhibited strongly by MnCl(2) and CuSO(4) at 1 mM and completely by FeSO(4) and Ag(2)SO(4) at 1 mM. A bacterial levan from Zymomonas mobilis was incubated with an LFTase; final conversion yield from the levan to DFA IV was 35%. Neither inulin, levanbiose, sucrose, dextran, nor starch was hydrolyzed by LFTase. DFA IV was very stable at acidic pH and high temperature, thus indicating that DFA IV may be suitable for the food industry and related areas.     

Trichoderma atroviride promotes growth and enhances systemic resistance to Diplodia pinea in radiata pine (Pinus radiata) seedlings

Root drench application of Trichoderma atroviride isolates R32, R33, R40 and R84 promoted the growth of potted radiata pine seedlings. After 6 weeks, seedlings treated with R33 and R84 had thicker stems and greater stem and root biomass (p < 0.05) than untreated controls. Treatment with R32 increased seedling root biomass whilst R40 increased stem diameter. None of the isolates affected seedling height. One isolate, R33, induced systemic resistance to stem inoculation with Diplodia pinea and reduced dieback incidence by 20% compared with untreated controls. To our knowledge, this is the first report of systemic induced resistance by Trichoderma in a pine species. Furthermore, seedlings that were treated with R33 (root drench) plus foliar application of methyl jasmonate (MeJA) expressed elevated peroxidase activity in their stems 2 weeks later, compared with seedlings treated only with MeJA. Because R33 itself did not affect peroxidase activity, this may be indicative of treatment synergy or defence potentiation by R33. Curiously, R33 + MeJA induced terpenoids but suppressed phenylalanine ammonia‐lyase activity suggesting possible trade‐offs between phenolic and terpenoid defence pathways in the treated seedlings.     

Mechanical properties of worsted fabrics for emotional garment to the rapier loom characteristics

This study examined the mechanical properties of worsted fabrics woven using various rapier weaving looms. For this purpose, the warp and weft yarn tensions during weaving were measured on the three types of rapier looms, and the fabric mechanical property changes due to the warp and weft tension differences were measured and analyzed according to the fabric position and particular rapier loom using the KES-FB system. The warp tension variation along the loom width direction in P-GTX loom showed the lowest value compared to FAST and THEMA looms. The warp tensions on the central part of the three types of looms were much higher than those on the left and right sides of the looms. The extensibility and bending rigidity of the fabric woven by P-GTX rapier loom showed lower values than those of FAST and THEMA looms, which appears to have originated from the low warp and weft weaving tensions of P-GTX rapier loom. On the other hand, the compressional property and shear modulus showed compromised results due to lateral deformation by compression and constraint deformation of the warp and weft by shear. The friction coefficient of the fabric surface woven by FAST loom showed the lowest value due to the flatter surface by the high warp tension. The mechanical properties of the fabric loaded by a high warp tension on the central part of the loom were also affected by the high weft yarn crimp due to the wider spacing between warp yarns by the higher warp tension during weaving, which makes the surface of the central part of the fabric flatter and smoother than the edge part of the fabric.     

Effectiveness of flavourzyme treatment on polyamide fabric

We characterized the effectiveness of Flavourzyme treatment in the hydrolysis of amide bonds in polyamide fabric by quantitating the ionic groups released into the treatment liquid and those formed on the fabric surface. On the basis of hydrolytic activity, we demonstrated that Flavourzyme effectively hydrolyzed amide bonds in polyamide (PA) fabric. The optimal treatment conditions were found to be pH 7.0, temperature 40 °C, treatment time 120 min, and Flavourzyme concentration 10 % based on weight of fiber. PA fabric treated with Flavourzyme exhibited increased numbers of amino and carboxyl groups, as evaluated by zeta potential and color strength. As the amounts of ionic groups formed by Flavourzyme hydrolysis increased, the water contact angle and water absorbency time decreased.     

Infection criteria,inoculum sources and splash dispersal pattern of <Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> causing bitter rot of apple in New Zealand

Infection of Malus x domestica cv. Royal Gala fruit by Colletotrichum acutatum causing bitter rot was studied in the temperate climate of New Zealand. Temperatures above 15 °C were required for lesions to develop on detached apple wound-inoculated or inoculated without wounding with C. acutatum spores, regardless of maturity. A wetness period of 72 h was required for infection of mature detached apple fruit without wounding. On wound-inoculated detached apple fruits, sporulation was related to temperature and followed a similar pattern. In the field, a mean temperature above 15 °C for 72 h after wound-inoculation was required for lesions to develop. Buds were a more important source of inoculum than twigs, and it was shown that C. acutatum could be isolated more frequently from outer bud scales than from inner scales. Asymptomatic infection of vegetative and reproductive buds was detected. C. acutatum was detected on asymptomic surface-sterilised petals and fruit, more commonly during summer than spring. Symptomless sterilised leaves generally yielded C. acutatum throughout the season, but isolations were more frequent in summer. Recovery of inoculum using a splash meter to detect vertical dispersal showed that in summer inoculum was primarily splashed up from the ground. In spring, inoculum was recovered in similar quantities from all heights up to a metre, suggesting that splash dispersal occurs from the canopy as well as from the ground. A disease cycle for C. acutatum infecting apples and causing bitter rot in New Zealand is suggested.