Ultrasonic Acoustic Emissions from the Sapwood of Thuja occidentalis Measured inside a Pressure Bomb

An improved method of counting acoustic emission (AE) events from water-stressed stems of cedar (Thuja occidentalis L.) is presented. Amplified AEs are analyzed on a real time basis by a microcomputer. The instrumentation counts AE events in a fashion nearly analogous to scintillation counting of radioactive materials.

The technique was applied to measuring ultrasonic AEs from the stems of cedar inside a pressure bomb. The shoots were originally fully hydrated. When the shoots are dehydrated in the bomb by application of an overpressure very few AEs were detected. When the bomb pressure is reduced after dehydration of the shoot, AE events could be detected. We conclude that ultrasonic AEs are caused by cavitation events (= structural breakdown of water columns in the tracheids of cedar) and not by the breaking of cellulose fibers in the wood.

     

Radial shrinkage and ultrasound acoustic emissions of fresh versus pre-dried Norway spruce sapwood

Acoustic emission (AE) and radial shrinkage were compared between fully saturated fresh and pre-dried Norway spruce sapwood during dehydration at ambient temperature. Hydraulic conductivity measurements, anatomical investigations on bordered pits and X-ray computed tomography (CT) scans were done to search for possible AE sources other than the breakage of the water columns inside the tracheids. Both fresh and pre-dried specimens showed radial shrinkage due to drying surface layers right from the beginning of dehydration, which induced almost no AE. Whereas no dimensional changes occurred in pre-dried wood thereafter, fresh wood showed a rapid shrinkage increase starting at 25% relative water loss. This dimensional change ceased when further moisture got lost and was even partially reversed. AE of fresh wood showed much higher activity and energy, which is a waveform feature that describes the strength of the acoustic signal. Extremely high single AE energy events were detected at this critical stage of dehydration. After partial recovery from shrinkage, neither dimensional changes nor AE activity showed differences between fresh and pre-dried wood after more than 80% relative moisture loss. Our results suggested that fresh sapwood is more prone to dehydration stresses than pre-dried sapwood. Differences in AE and shrinkage behavior might be due to the weakening or distortion of the pit membranes (cavitation fatigue), pit aspiration, structural changes of the cell walls and micro-checks, which occurred during the first dehydration cycle.     

Water relation parameters of embryogenic cultures and seedlings of larch

Changes in the water relations parameters of developing somatic embryogenic and xygotic European larch (Larix decidua) were studied. Water release curves were generated by suspending tissue samples over unsaturated NaCl solutions until they reached vapor equilibration with the surrounding air. Twenty solutions were used whose water potentials ranged from −0.05 to −10 MPa. Water release curves were obtained by plotting paired values of tissue relative water content (RWC) and solution potential. Curves were derived for embryonic larch at various stages of development and for hypocotyls and roots from germinated zygotic and somatic embryos. The ability to resist dehydration increased markedly with development. Stage 1 tissue, which consisted of clusters of loosely associated nonchlorophyllous cells, had extremely low bulk elastic modulus (ε) (1.91 MPa) and apoplastic water content (A) (0.023), relatively high osmotic potential (Ψ_π) (−0.53 MPa), and lost turgor at 0.56 RWC. In contrast, mature embryoids with primary roots, hypocotyl, and cotyledons (stage 3) had an almost 4-fold increase in A (0.089), significantly higher ε (3.49 MPa), and lower Ψ_π (−0.88 MPa) and lost turgor at 0.66 RWC. Hypocotyl tissue from germinated somatic embryos lost turgor at 0.74 RWC and had higher ε, A, and solute accumulation than pregerminated tissue. Hypocotyl tissue resisted dehydration more strongly than root tissue, and differences between root and hypocotyl water relation parameters were more pronounced in xygotic than in somatic seedlings. Highest dehydration resistance was in zygotic hypocotyls. The characterization of the water relations of tissue cultures should allow the development of more consistent and reliable desiccation protocols to induce maturation of embryos and produce synchronously germinating seed.     

Detection of xylem cavitation in field-grown pine trees using the acoustic emission technique

The relationship between the ultrasonic acoustic emission (AE) and the needle water potential in field-grown Japanese black pine,Pinus thunbergii, and excised-pine shoots was investigated in summer. The AE technique was employed as a reliable indicator of cavitation events in the water columns of xylem tracheids even under field conditions. In excised shoots. AE events detected when the needle water potential fell below about-0.9 MPa. In the case of field-grown pine trees, however, relatively few AE event were detected when needle water potential was lower than-0.9 MPa. This suggests that the occurrence of cavitation events in Japanese black pine growing under field conditions is comparatively rare, even in summer.     

Cellular level observation of water loss and the refilling of tracheids in the xylem of Cryptomeria japonica during heartwood formation

The mechanism of heartwood formation in Cryptomeria japonica D. Don has long been studied since heartwood formation is a fundamental physiological feature of trees. In this study, the water distribution in the xylem of C. japonica was investigated at the cellular level to reveal the role of water distribution in the xylem during heartwood formation. Samples were taken from different heights of each trunk, in which the phases of heartwood formation differed. These were designated as SIH, which consisted of sapwood, intermediate wood, and heartwood; SI, which consisted of sapwood and intermediate wood but no heartwood; and S-all, which consisted entirely of sapwood. Cryo-scanning electron microscopic observations of the heartwood-formed (SIH) and non-heartwood-formed (SI and S-all) xylem revealed different patterns of water distribution changes in tracheids between the latewood and earlywood. In the latewood, almost all tracheids were filled with water in all areas from the sapwood to the heartwood (98–100% of tracheids had water in their lumina). In the earlywood, however, the water distribution differed between the sapwood (95–99%), intermediate wood (7–12%), and heartwood (4–100%). Many of the tracheids in the xylem, where the sapwood changed to intermediate wood lost water. In the heartwood, some tracheids remained empty, while others were refilled with water. These results suggest that the water distribution changes in individual tracheids are closely related to heartwood formation. Water loss from tracheids may be an important factor inducing heartwood formation in the xylem of C. japonica.     

Morphology of the perennial ryegrass (Lolium perenne; Poaceae) coleorhiza and emerging radicle with continuous or discontinuous hydration

This study was conducted using scanning electron microscopy to characterize morphological changes in the maturing perennial ryegrass (Lolium perenne L.) coleorhiza and emerging radicle during continuous hydration or hydration interrupted by dehydration. With continuous hydration, coleorhizae emerged and cell expansion led to a progressive increase in tissue size. Coleorhiza cells developed extensions like epidermal root hairs. Although coleorhiza cells appeared undamaged by radicle emergence, they began deteriorating when the radicle had reached a length of approximately 2 mm. In response to dehydration, coleorhiza cells shrank but did not rupture. If dehydration was initiated during an early stage of coleorhiza development, greater tissue shrinkage occurred at -150 MPa than at -4 MPa; otherwise, coleorhizae showed no differential response due to dehydration water potential. Upon rehydration, coleorhizae dehydrated at -4 MPa regained cell turgor within 24 hr, while coleorhizae dehydrated at -150 MPa did not. Loss of the coleorhiza (due to desiccation) did not preclude radicle emergence, which occurred upon rehydration. Radicles up to 2 mm in length were more tolerant of dehydration than were coleorhizae. These results suggest that the coleorhiza may be an expendable tissue during germination, as its likely protective and absorptive roles are lost following a single harsh dehydration event.     

Cavitation Events in Thuja occidentalis L.? : Utrasonic Acoustic Emissions from the Sapwood Can Be Measured

Ultrasonic acoustic emissions (AE) in the frequency range of 0.1 to 1 megahertz appear to originate in the sapwood of Thuja occidentalis L. The AE are vibrations of an impulsive nature. The vibrations can be transduced to a voltage waveform and amplified. The vibrations of each AE event begin at a large amplitude which decays over 20 to 100 microseconds. Strong circumstantial evidence indicates that the ultrasonic AE result from cavitation events because: (a) they occur only when the xylem pressure potential Ψ_xp is more negative than a threshold level of about —1 megapascal; (b) the rate of AE events increases as Ψ_xp decreases and when the net rate of water loss increases; (c) the AE can be stopped by raising Ψ_xp above —1 megapascal. Ultrasonic AE have been measured in whole terminal shoots allowed to dry in the laboratory, in isolated pieces of sapwood as they dried in the laboratory, and in whole terminal shoots in a pressure bomb when Ψ_xp was decreased by lowering the gas pressure in the pressure bomb.     

Seasonal and perennial changes in the distribution of water in the sapwood of conifers in a sub-frigid zone

An analysis was made of progressive changes in patterns of cavitation in the sapwood of three species of conifer (Larix kaempferi, Abies sachalinensis, and Picea jezoensis) that were growing in a sub-frigid zone. In all three conifers, all tracheids of the newly forming outermost annual ring were filled with water or cytoplasm during the period from May to August. However, many tracheids in the transition zone from earlywood to latewood lost water in September, presumably through drought-induced cavitation. Cavitated tracheids tended to be continuously distributed in a tangential direction. Subsequently, some earlywood tracheids of the outermost annual ring lost water during the period from January to March. This was associated with freeze-thaw cycles. In the second and third annual rings from the cambium of all three conifers, the lumina of most tracheids in the transition zone from earlywood to latewood contained no water. In contrast, some latewood tracheids near the annual ring boundary and many earlywood tracheids retained water in their lumina. The third annual ring had more cavitated tracheids than the second annual ring. Our observations indicated that cavitation progressed gradually in the tracheids of the conifers and that they were never refilled once cavitation had occurred. The region involved in water transport in conifers did not include the entire sapwood and differed among annual rings.     

Collapse of Water-Stress Emboli in the Tracheids of Thuja occidentalis L

We report the kinetics of embolus formation and collapse in the tracheids of Thuja occidentalis L. stem segments. Radial wood sections were trimmed to 4 mm long paralleling the tracheids by 1 mm wide and 0.1 mm thick. They were observed under a dissecting microscope at 128x while sections were dehydrated and rehydrated. During dehydration, cavitations resulted in the formation of emboli in tracheids, but we concluded that the cavitated tracheids did not immediately fill with air at atmospheric pressure. This conclusion was based on the time required for the emboli to collapse after the rewetting of the dehydrated segment. By hypothesis, the time for the emboli to collapse should be proportional to the amount of air in the emboli. The time for all the emboli to collapse was a linear function of the dehydration time for times up to 15 min. For dehydrations greater than 80 min, the time for collapse after rewetting was constant, and we concluded that the tracheids have saturated with air by 80 min of dehydration. The kinetics of embolus formation is discussed in terms of the air-seeding hypothesis for cavitation, and collapse is discussed in terms of the physics of gas dissolution and diffusion. Embolus formation and dissolution in intact herbaceous and woody plants should follow the same physical laws.     

影响蝴蝶兰类原球茎耐脱水性的主要因素

为探讨蝴蝶兰(Phalaenopsis spp.)类原球茎(protocorm-like body,PLB)耐脱水性的主要影响因素,对PLB的平均粒重、含水率、脱水相对湿度、时间、温度、光周期与耐脱水性的关系进行了研究.结果表明,PLB的平均粒重与脱水后失水率、含水率、相对电导率、成活率呈显著或极显著相关.在较高湿度下...     

The Measurement and Interpretation of Ultrasound from Woody Stems

A system for detecting and counting ultrasonic acoustic emissions(AEs) from woody stems in the waveband of 0.1–1.0 MHzis described. AEs from small pieces of stems of Mains sylvestrisand Chamaecyparis lawsoniana occurred up to a rate of 6000 min^–1,and could be related to the water loss as the stems dried. AEsdid not occur randomly, but were aggregated. Each event is interpretedas a single cavitation occurring in the water within a tracheidor fibre. AEs could be temporarily stopped by rewetting thetissue or cutting it to release tension. From knowledge of thenumber of cells in the stems, the fraction of total counts pernumber of tracheids or fibres present in the stem was calculated.This was found to be less than 0.16. One reason why this fractionis so low is that AEs may occur simultaneously; it is also probablethat some AEs emit energy outside the waveband studied. Eventhough the fraction of AEs recorded is low, this instrumentcould prove to be a very powerful tool for investigating theoccurrence of cavitation in woody stems. Key words: Ultrasound, Cavitation, Wood     

Embolism formation during freezing in the wood of Picea abies

Freeze-thaw events can cause embolism in plant xylem. According to classical theory, gas bubbles are formed during freezing and expand during thawing. Conifers have proved to be very resistant to freeze-thaw induced embolism, because bubbles in tracheids are small and redissolve during thawing. In contrast, increasing embolism rates upon consecutive freeze-thaw events were observed that cannot be explained by the classical mechanism. In this study, embolism formation during freeze-thaw events was analyzed via ultrasonic and Cryo-scanning electron microscope techniques. Twigs of Picea abies L. Karst. were subjected to up to 120 freeze-thaw cycles during which ultrasonic acoustic emissions, xylem temperature, and diameter variations were registered. In addition, the extent and cross-sectional pattern of embolism were analyzed with staining experiments and Cryo-scanning electron microscope observations. Embolism increased with the number of freeze-thaw events in twigs previously dehydrated to a water potential of -2.8 MPa. In these twigs, acoustic emissions were registered, while saturated twigs showed low, and totally dehydrated twigs showed no, acoustic activity. Acoustic emissions were detected only during the freezing process. This means that embolism was formed during freezing, which is in contradiction to the classical theory of freeze-thaw induced embolism. The clustered pattern of embolized tracheids in cross sections indicates that air spread from a dysfunctional tracheid to adjacent functional ones. We hypothesize that the low water potential of the growing ice front led to a decrease of the potential in nearby tracheids. This may result in freezing-induced air seeding.     

Corticular photosynthesis drives bark water uptake to refill embolized vessels in dehydrated branches of Salix matsudana

It is well known that xylem embolism can be repaired by bark water uptake and that the sugar required for embolism refilling can be provided by corticular photosynthesis. However, the relationship between corticular photosynthesis and embolism repair by bark water uptake is still poorly understood. In this study, the role of corticular photosynthesis in embolism repair was assessed using Salix matsudana branch segments dehydrated to ?1.9 MPa (P₅₀, water potential at 50% loss of conductivity). The results indicated that corticular photosynthesis significantly promoted water uptake and nonstructural carbohydrate (NSC) accumulation in the bark and xylem during soaking, thereby effectively enhancing the refilling of the embolized vessels and the recovery of hydraulic conductivity. Furthermore, the influence of the extent of dehydration on the embolism refilling enhanced by corticular photosynthesis was investigated. The enhanced refilling effects were much higher in the mildly dehydrated (?1.5 MPa) and moderately dehydrated (?1.9 MPa) branch segments than in the severely dehydrated (?2.2 MPa) branch segments. This study provides evidence that corticular photosynthesis plays a crucial role in xylem embolism repair by bark water uptake for mildly and moderately dehydrated branches.     

Characterization and propagation of acoustic emission signals in woody plants: towards an improved acoustic emission counter

Abstract. The physics of ultrasonic acoustic emissions (AEs) was investigated for AE transmission through wood and transducers. The physical properties measured were velocity, attenuation and frequency composition of AEs produced by two sources: cavitation events in xylem and pencil lead breaks. The authors also measured the relative sensitivity of various combinations of ultrasound transducers and amplifiers to aid in the selection of a measuring system optimized for cavitation detection in woody plants. Some of the authors' conclusions are: (1) Softwoods ( Thuja, Pinus ) attenuate AEs more rapidly than hardwoods (maple, birch). (2) The velocity of AEs in wood exceeds that measured by others in water so the main medium of AE transmission must be the cellulose. (3) The strongest frequencies of AEs are in the range of 100–300 kHz. (4) Cavitation-induced AEs tend to shift to higher frequency as wood dehydration progresses. (5) One cannot determine the locus of origin of AEs from its frequency composition. (6) The frequency composition of the acoustic emissions probably cannot be determined at all with the sensors used because of their tendency to 'ring'. The data collected in this paper were used to aid in the design of an improved AE counter having a seven-fold increase in signal to noise ratio compared to counters previously used in our laboratory. The improved counter, model 4615 Drought Stress Monitor, is now commercially available from Physical Acoustics Corp., Princeton, NJ, U.S.A.     

Temperature and water content effects on the viscoelastic behavior of<Emphasis Type="Italic"> Tilia americana</Emphasis> (Tiliaceae) sapwood

The effects of temperature and water content on the viscoelasticity of living and dehydrated Tilia americana sapwood were examined using transient creep (time- and load-dependent deformation) tests under sustained bending loads. Creep tests were performed at 21.1°C and –20.5°C to determine the magnitudes and types of strains in living and dehydrated samples. Temperature had no effect on the creep rate of living sapwood. However, the creep rate of dehydrated samples at –20.5°C was significantly faster than that at 21.1°C. Regardless of temperature, sapwood had a faster creep rate than dehydrated samples. With small bending loads, the residual strains in sapwood were larger at 21.1°C compared to –20.5°C. Temperature did not significantly affect the residual strains in dehydrated samples. For small bending loads, frozen sapwood recovered all residual creep strains when thawed. With larger loads, residual and plastic (permanent) strains increased. We speculate that ice formation in cell lumens partially dehydrates (and thus stiffens and strengthens) cell wall materials and prevents cell wall buckling and elastic restoration after unloading. However, when thawed, sapwood can elastically restore its original configuration, provided it is not excessively bent (by ice or snow accumulations) when frozen.     

Water balance and osmoregulation in Onymacris plana, a tenebrionid beetle from the Namib desert

Onymacris plana, a tenebrionid beetle from the sand dunes of the Namib desert, lost weight very slowly during 12 days of dehydration at 26°C. Measurement of total lipid showed a gradual decline, the metabolic water produced being sufficient to maintain a constant water content. At the same time the haemolymph volume decreased by 66%. When given water the dehydrated beetles drank rapidly and their weight and haemolymph volume were restored to normal. Haemolymph osmolarity was closely regulated despite the changes in volume. Haemolymph potassium was also well regulated, but sodium was lost from the haemolymph during a cycle of dehydration and rehydration, even though sodium losses in the faeces were small. Water balance in Onymacris depends on efficient conservation of water in periods of drought and on water uptake by drinking during the coastal fogs of the Namib.     

The Limits to Xylem Embolism Recovery in Pinus sylvestris L.

In this study we test the hypothesis that, when water supplyis under tension, reversal of cavitation can occur as long aswater continuity is maintained in the vicinity of tracheids.The experiments were conducted on young branches, 7–8mm diameter, of Pinus sylvestris L., freshly collected and allowedto lose water on the bench after being debarked. During dehydration,the volumetric fractions of water (V_w) and gas (V_s) changedsteadily as relative water content () declined. Meanwhile, ultrasonicemissions (UAE) started after a threshold = 90% was reachedand were maximal at = 75%. Before and after dehydration, branchsegments were connected to water-filled tubing and placed from0.2 to 3.6 m above a water source and water inflow and outflowwere recorded. These distances provided a source of water ata potential of –2.0 to –36kPa. We considered thatthe segment water potential would be a function of the surfacetension across the water meniscii at the ends of the embolizedtracheids. Thus, water potentials calculated from tracheid dimensionswould be as low as –43 kPa. Water inflow to segments declinedwhen the distance from the source was increased or the segmentswere very dehydrated. Increasing the distance above the watersource would be expected to increase the water potential differencebut to reduce water uptake. The most dehydrated segments absorbedwater faster at the beginning of the refilling period (2h),but at the end of 16h, was lower and V_g larger than in lessembolized tissue. Recovery of water flow followed a similartrend, and was lowest when embolisms increased. For a narrowrange of , hydraulic conductance was reduced sharply, indicatingthat wide tracheids were still gas-filled. Thus, the numberof tracheids remaining embolized increased when the source waterpotential was low and there was severe embolism. We concludethat embolism can be reversed in P. sylvestris at a rate dependingon the water potential of the source, severity of embolism andhydraulic conductivity. Key words: Pinus sylvestris L., cavitation induction-recovery, embolized tracheids, water content and embolism, matrix potential, capillary, hydraulic conductance     

The function of intercellular spaces along the ray parenchyma in sapwood, intermediate wood, and heartwood of Cryptomeria japonica (Cupressaceae)

? Premise of the study: Intercellular spaces along ray parenchyma (ISRP) are common in many conifer xylems, but their function is uncertain because the in-situ structural network among ISRP, ray parenchyma, and tracheids has not been evaluated. Analysis of water distribution in ISRP from sapwood to heartwood is needed to elucidate the function of ISRP in sapwood, intermediate wood, and heartwood. ? Methods: We used cryo-scanning electron microscopy, x-ray photography, and water content measurement in xylem to analyze the presence of liquids in ISRP, ray parenchyma, and tracheids from sapwood to heartwood in Cryptomeria japonica (Cupressaceae). ? Key results: In sapwood, almost all ISRP were empty. "Cingulate-cavitated regions", which lose water along the tangential direction within one annual ring, formed in the earlywood tracheids, and their frequency increased toward the inner annual rings, whereas ray parenchyma cells were alive and not involved in the partial cavitation. In intermediate wood, almost all ISRP and earlywood tracheids and many of the ray cells were empty, and only some latewood tracheids retained liquid in their lumina. The ISRP were connected with tracheids via gas-filled ray parenchyma cells. ? Conclusions: The ISRP work as a pathway of gas for aspiration of ray parenchyma cells in sapwood. On the other hand, the occurrence of a gas network between ISRP, ray parenchyma, and tracheids facilitates cavitation of tracheids, resulting in the generation of low-moisture, intermediate wood.     

Safety and tolerability of once-daily umeclidinium/vilanterol 125/25 mcg and umeclidinium 125 mcg in patients with chronic obstructive pulmonary disease: results from a 52-week,randomized, double-blind,placebo-controlled study

Background

The long-acting muscarinic antagonist (LAMA) umeclidinium (UMEC) and the combination of UMEC with the long-acting β₂-agonist (LABA) vilanterol (UMEC/VI) are approved maintenance treatments for chronic obstructive pulmonary disease (COPD) in the US and EU. They are not indicated for the treatment of asthma.

Methods

In this 52-week, double-blind, placebo-controlled, parallel-group safety study (GSK study DB2113359; {"type":"clinical-trial","attrs":{"text":"NCT01316887","term_id":"NCT01316887"}}NCT01316887), patients were randomized 2:2:1 to UMEC/VI 125/25 mcg, UMEC 125 mcg, or placebo. Study endpoints included adverse events (AEs), clinical chemistry and hematology parameters, vital signs, 12-lead, and 24-hour Holter electrocardiograms. COPD exacerbations and rescue medication use were assessed as safety parameters; lung function was also evaluated.

Results

The incidence of on-treatment AEs, serious AEs (SAEs), and drug-related AEs was similar between treatment groups (AEs: 52–58%; SAEs: 6–7%; drug-related AEs: 12–13%). Headache was the most common AE in each treatment group (8–11%). AEs associated with the LAMA and LABA pharmacologic classes occurred at a low incidence across treatment groups. No clinically meaningful effects on vital signs or laboratory assessments were reported for active treatments versus placebo. The incidences of atrial arrhythmias with UMEC/VI 125/25 mcg were similar to placebo; for UMEC 125 mcg, the incidences of ectopic supraventricular beats, sustained supraventricular tachycardia, and ectopic supraventricular rhythm were ≥2% greater than placebo. With active treatments, COPD exacerbations were fewer (13–15% of patients reporting ≥1 exacerbation) and on average less rescue medication was required (1.6–2.2 puffs/day) versus placebo (24% reporting ≥1 exacerbation, 2.6 puffs/day). Both active treatments improved lung function versus placebo.

Conclusion

UMEC/VI 125/25 mcg and UMEC 125 mcg were well tolerated over 12 months in patients with COPD.