IDENTIFICATION OF ENDOPHYTIC BACTERIA ASSOCIATED WITH N2 FIXATION AND INDOLE ACETIC ACID SYNTHESIS AS GROWTH PROMOTERS IN CURCUMA ALISMATIFOLIA GAGNEP.

Endophytic bacteria carrying out dinitrogen (N₂) fixation and indole acetic acid (IAA) synthesis were firstly identified in C. alismatifolia, a globally important flower crop. Their potential as growth promoters to stimulate the rapid growth of host plant was also examined. It will be beneficial to reduce the propagation period of tissue culture plantlets, and also utilize as a biofertilizer for rhizome production in the field. Seven endophytic bacteria were isolated from the leaf, four isolates from the leaf base, and two from the rhizome. ECS203, a gram-negative bacterium with a round shape, showed the highest N₂ fixation at 4.2 nmol C₂H₄/10⁶ cells/hr, and ECS202 showed the highest IAA synthesis at 296 μL μg ^{? 1} protein. Three selected isolates of N₂-fixing and IAA synthesizing endophytic bacteria, i.e., ECS202, ECS203, and ECS204, isolated from the leaf base, were used to reinoculate Curcuma plantlets derived from tissue culture. Then, plants were grown in sterilized sand for 2 months and weekly supplied with N-free nutrient solution. Plant growth, colonization, nitrogen fixation, and IAA synthesis were measured at two months after planting. The inoculated plants clearly showed a better performance of plant growth and yield in terms of the plant height, plant weight, leaf area, and diameter of new rhizomes compared with uninoculated plants. The chlorophyll content and N concentration of leaves and roots also increased in inoculated plants. Endophytic bacteria from inoculated plants colonized the roots, rhizome, and leaf base. Partial sequence analysis using 16S rDNA indicated that the isolate ECS202 corresponded to Sphingomonas pseudosanguinis (99.2% similarity over 1,371 bp), ECS203 to Bacillus drentensis (99.4% similarity over 1,450 bp) and ECS204 to Bacillus methylotrophicus (99.9% similarity over 13,06 bp).     

Sublingual administration of detomidine to calves prior to disbudding: a comparison with the intravenous route

ObjectiveTo study the effects of oromucosal detomidine gel administered sublingually to calves prior to disbudding, and to compare its efficacy with intravenously (IV) administered detomidine.Study designRandomised, prospective clinical study.AnimalsTwenty dairy calves aged 12.4 ± 4.4days (mean ± SD), weight 50.5 ± 9.0 kg.MethodsDetomidine at 80 μg kg^?1 was administered to ten calves sublingually (GEL) and at 30 μg kg^?1 to ten control calves IV (V. jugularis). Meloxicam (0.5 mg kg^?1) and local anaesthetic (lidocaine 3 mg kg^?1) were administered before heat cauterization of horn buds. Heart rate (HR), body temperature and clinical sedation were monitored over 240 minutes. Blood was collected from the V. cephalica during the same period for drug concentration analysis. Pharmacokinetic variables were calculated from the plasma detomidine concentration-time data using non-compartmental methods. Statistical analyses compared routes of administration by Student’s t-test and linear mixed models as relevant.ResultsThe maximum plasma detomidine concentration after GEL was 2.1 ± 1.2 ng mL^?1 (mean ±SD) and the time of maximum concentration was 66.0 ± 36.9 minutes. The bioavailability of detomidine was approximately 34% with GEL. Similar sedation scores were reached in both groups after administration of detomidine, but maximal sedation was reached earlier in the IV group (10 minutes) than in the GEL group (40 minutes). HR was lower after IV than GEL from 5 to 10 minutes after administration. All animals were adequately sedated, and we were able to administer local anaesthetic without resistance to all of the calves before disbudding.Conclusions and clinical relevanceOromucosally administered detomidine is an effective sedative agent for calves prior to disbudding.     

Plasma drug concentrations and clinical effects of a peripheral alpha‐2‐adrenoceptor antagonist,MK‐467, in horses sedated with detomidine

ObjectiveTo investigate plasma drug concentrations and the effect of MK-467 (L-659′066) on sedation, heart rate and gut motility in horses sedated with intravenous (IV) detomidine.Study designExperimental randomized blinded crossover study.AnimalsSix healthy horses.MethodsDetomidine (10 μg kg^?1 IV) was administered alone (DET) and in combination with MK-467 (250 μg kg^?1 IV; DET + MK). The level of sedation and intestinal sounds were scored. Heart rate (HR) and central venous pressure (CVP) were measured. Blood was collected to determine plasma drug concentrations. Repeated measures anova was used for HR, CVP and intestinal sounds, and the Student's t-test for pairwise comparisons between treatments for the area under the time-sedation curve (AUC_sed) and pharmacokinetic parameters. Significance was set at p < 0.05.ResultsA significant reduction in HR was detected after DET, and HR was significantly higher after DET + MK than DET alone. No heart blocks were detected in any DET + MK treated horses. DET + MK attenuated the early increase in CVP detected after DET, but later the CVP decreased with both treatments. Detomidine-induced intestinal hypomotility was prevented by MK-467. AUC_sed was significantly higher with DET than DET + MK, but maximal sedations scores did not differ significantly between treatments. MK-467 lowered the AUC of the plasma concentration of detomidine, and increased its volume of distribution and clearance.Conclusions and clinical relevanceMK-467 prevented detomidine induced bradycardia and intestinal hypomotility. MK-467 did not affect the clinical quality of detomidine-induced sedation, but the duration of the effect was reduced, which may have been caused by the effects of MK-467 on the plasma concentration of detomidine. MK-467 may be useful clinically in the prevention of certain peripheral side effects of detomidine in horses.     

Improving productivity of four fruit species by growth regulators applied once in ultra-low doses to the collar

Summary

Two growth retardants: paclobutrazol (P, Cultar) or 2,3,5-triiodobenzoic acid (TIBA) were applied once alone, or in mixtures (in a range of 5 to 20 mg per tree) to the collar of maiden plum, sour and sweet cherries and apple trees, in early spring of the second year after planting. Plum and apple trees were also treated with benzyladenine (BA) at 10 mg per tree a.i. in a mixture with P or TIBA. Sweet and sour cherry trees were treated with natural phenolic substances: phloridzin (Phi) and quercetin (Que) alone, or in mixtures with P and TIBA. A mid-stem treatment with P and shoot bending were also applied to the plum and apple trees for comparison. Measurements of tree growth and fruiting were made within 4 or 5 years. The reaction of the four species to the treatments varied according to the growth regulator applied. Plum trees responded mostly to TIBA and its mixtures with P. A strong suppression of tree growth and increased fruit productivity, as well as improved fruit quality, were observed. The TIBA application and its mixtures with P were also effective in causing growth reduction of the sweet cherry trees. Treatments with P alone, or mixed with TIBA, were effective in growth limitation of sour cherry trees. Some increase in the reproductive processes was observed only after the TIBA treatment in both species. The apple trees responded to application of mixtures of TIBA with benzyladenine (BA), or with P, and to P alone, with effective growth reduction. But only the P + BA treatment increased significantly the fruiting of apple trees, while the other treatments resulted in crops proportional to the diminished tree sizes. The mid-stem treatment did not affect plum trees but increased growth of apple trees. Shoot bending had no effect on the plum trees but increased fruiting of apple trees. The addition of Phi to half the lower P dose or TIBA, magnified growth suppression in the sweet and sour cherry trees. When applied alone Que caused a small growth inhibition but Phi produced some increase in growth of sour cherry trees. Results obtained show the possibilities of practical applications of growth regulators to the collar. Their reduced doses mixed with natural phenolic substances are equally effective in growth suppression and make fruit production safer and more profitable, especially in plum and sour cherry trees.     

Application of the wood properties of large-diameter Sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) logs to sorting logs and sawing patterns

The purposes of this study were to accumulate fundamental data on wood properties within large Sugi logs and to take applicable variations in wood properties into consideration for sorting logs and sawing patterns. The characteristics of basic density, moisture content, growth ring width, and microfibril angle (MFA) were measured and the relationship with log and lumber quality was examined. It was considered reasonable to estimate the lumber moisture content based on the moisture content of heartwood rather than that of whole logs, especially when producing large-sized lumber. The MFA reached a constant value before the 15th ring, and within a distance of 10 cm or less from the pith. Since the E _fr of lumber correlated with that of the log affected by MFA, it would be possible to produce lumber with a higher E _fr from the outer position of the log, based on selecting a log above the E _fr . Since the MFA would also affect the lumber warp, a sawing pattern avoiding the area around the pith or enlarging the rough sawn size when a large warp was expected could be effective in improving the lumber quality. To improve the lumber quality, not only one but also multiple wood properties must be applied to the sawing pattern.     

The effects of early and intense pruning on light penetration,tree growth,and epicormic shoot dynamics in a young hybrid larch stand

We examined the effect of early and intense pruning on light intensity under the canopy, individual growth, diameter–height relationships, and epicormic shoot dynamics in young hybrid larch (Larix gmelinii var. japonica × L. kaempferi) to establish a new effective management method for hybrid larch plantations. The objective is to produce high-quality wood while reducing silviculture costs using a combination of low-density planting and early and intense pruning. In a young hybrid larch plantation, we pruned branches to two different heights (2 and 4 m above ground level) using a no-pruning treatment as a control. Although the growth rates were lower in the heavy pruning treatment (4 m above the ground level) than in other treatments in the year following pruning, when measured 4 years later, growth did not differ between treatments. The number of epicormic shoots increased in the year following pruning, as did the relative photosynthetic photon flux density (rPPFD). The number of epicormic shoots was also dependent on the size of individual trees. However, survival of epicormic shoots was not sufficiently high to be problematic for high-quality timber production. If branches are pruned carefully such that the rPPFD does not rise above 20%, the emergence of epicormic shoots can also be controlled. Our results indicate that early and intense pruning is an effective component of a new management system for hybrid larch plantations.     

Myosin substitution rate is affected by the amount of cytosolic myosin in cultured muscle cells

In striated muscles, approximately 300 myosin molecules form a single thick filament in myofibrils. Each myosin is continuously displaced by another myosin to maintain the thick filament structure. Our previous study using a fluorescence recovery after photobleaching (FRAP) technique showed that the myosin replacement rate is decreased by inhibition of protein synthesis, but myosin is still exchangeable. This result prompted us to examine whether myosin in the cytoplasm is involved in myosin replacement in myofibrils. To address this, FRAP was measured in green fluorescent protein (GFP)‐tagged myosin heavy chain 3 (Myh3) expressing myotubes that were treated with streptolysin‐O (SLO), which forms pores specifically in the plasma membrane to induce leakage of cytoplasmic proteins. Our biochemical data demonstrated that the cytoplasmic myosin content was reduced in SLO‐permeabilized semi‐intact myotubes. Furthermore, FRAP experiments showed a sluggish substitution rate of GFP‐Myh3 in SLO‐permeabilized myotubes. Taken together, these results demonstrate that the myosin substitution rate is significantly reduced by a decreased amount of myosin in the cytoplasm and that cytoplasmic myosin contributes to myosin replacement in myofibrils.     

Evaluation of the polymorphonuclear cell functions of bottlenose dolphins

The functions of polymorphonuclear cells (PMN) are the important non-specific defense mechanisms in the immune system. Especially marine mammals are protected by these mechanisms from the aquatic environment with a large variety of microorganisms. Therefore, we examined the PMN functions of bottlenose dolphins in order to obtain the normal ranges and to standardize the techniques. PMNs were isolated by using lymphocyte isolate solution whose density was 1.077; superoxide production was assessed by nitroblue tetrazolium reduction test (NBT) and phagocytosis was tested by using polystyrene latex beads. We showed that the optimal incubation time was 30 min in NBT assay and 12 hr in phagocytosis assay for dolphin PMNs.     

A novel furostanol saponin from Tribulus terrestris of Bulgarian origin

The phytochemical investigation of the aerial parts of Tribulus terrestris of Bulgarian origin has resulted in the isolation of the novel furostanol saponin 1, named tribol, together with the known spirostanol saponins 2 and 3 and sitosterol glucoside. The structure of tribol was determined as (25R)-furost-5(6)-ene-3beta,16,26-triol-3-O-alpha-rhamnopyranosyl-(1-->2)-[alpha-rhamnopyranosyl-(1-->4)]-beta-glucopyranoside (1) by spectral analysis, including extensive 1D and 2D-NMR experiments.     

Comparison of changes in urinary and blood levels of biomarkers associated with proximal tubular injury in rat models

To investigate useful biomarkers associated with proximal tubular injury, we assessed changes in levels of a focused set of biomarkers in urine and blood. Male rats administered a single dose or four doses of gentamicin (GM, 240 mg/kg/day) or a single dose of cisplatin (CDDP, 5 mg/kg) were euthanized on days 2 (the day after initial dosing) 5, or 12. At each time point, histopathological examination of the kidney and immunohistochemistry for biomarkers, kidney injury molecule-1 (Kim-1), lipocalin (NGAL), clusterin (CLU), cystatin C (CysC) and β2-microglobulin (β2M) were performed. Biomarker levels were measured in urine and blood. In both treatment groups, degenerated/necrotic proximal tubules and regenerated tubules were mainly observed on days 5 and 12, respectively. At the same time as these tubular injuries, urinary Kim-1, CysC and β2M levels were increased. Moreover, urinary levels of CysC and β2M in GM-treated animals and Kim-1 in CDDP-treated animals increased (on day 2) prior to tubular injury on day 5. This was considered to reflect the characteristics of drug toxicity. Although almost all of the biomarkers in blood were not sufficiently sensitive to detect proximal tubular injury, urinary and plasma β2M levels simultaneously increased. Therefore, in addition to urinary Kim-1, CysC and β2M levels, plasma β2M levels were also considered useful for detecting proximal tubular injury.