视觉诱发电位波形的自动判别分析及其临床应用

计算诱发电位波形第一至第六次谐波的幅度和相位,把这12个参数作为判别分析的自变量.用32只正常眼、31只弱视眼和30只球后视神经炎患眼的诱发电位波形数据建立判别分析系统,再用另外32只正常眼、35只弱视眼和30只球后视神经炎患眼的诱发电位波形数据检验该判别分析系统的判别效果.     

High‐throughput screening techniques for rapid PEG‐based precipitation of IgG4 mAb from clarified cell culture supernatant

Locating optimal protein precipitation conditions for complex biological feed materials is problematic. This article describes the application of a series of high‐throughput platforms for the rapid identification and selection of conditions for the precipitation of an IgG₄ monoclonal antibody (mAb) from a complex feedstock using only microliter quantities of material. The approach uses 96‐microwell filter plates combined with high‐throughput analytical methods and a method for well volume determination for product quantification. The low material, time and resource requirements facilitated the use of a full factorial Design of Experiments (DoE) for the rapid investigation into how critical parameters impact the IgG₄ precipitation. To aid the DoE, a set of preliminary range‐finding studies were conducted first. Data collected through this approach describing Polyethylene Glycol (PEG) precipitation of the IgG₄ as a function of mAb concentration, precipitant concentration, and pH are presented. Response surface diagrams were used to explore interactions between parameters and to inform selection of the most favorable conditions for maximum yield and purification. PEG concentrations required for maximum yield and purity were dependant on the IgG₄ concentration; however, concentrations of 14 to 20% w/v, pH 6.5, gave optimal levels of yield and purity. Application of the high‐throughput approach enabled 1,155 conditions to be examined with less than 1 g of material. The level of insights gained over such a short time frame is indicative of the power of microwell experimentation in allowing the rapid identification of appropriate processing conditions for key bioprocess operations. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Computational Intelligence and Wavelet Transform Based Metamodel for Efficient Generation of Not-Yet Simulated Waveforms

The design and verification of complex electronic systems, especially the analog and mixed-signal ones, prove to be extremely time consuming tasks, if only circuit-level simulations are involved. A significant amount of time can be saved if a cost effective solution is used for the extensive analysis of the system, under all conceivable conditions. This paper proposes a data-driven method to build fast to evaluate, but also accurate metamodels capable of generating not-yet simulated waveforms as a function of different combinations of the parameters of the system. The necessary data are obtained by early-stage simulation of an electronic control system from the automotive industry. The metamodel development is based on three key elements: a wavelet transform for waveform characterization, a genetic algorithm optimization to detect the optimal wavelet transform and to identify the most relevant decomposition coefficients, and an artificial neuronal network to derive the relevant coefficients of the wavelet transform for any new parameters combination. The resulted metamodels for three different waveform families are fully reliable. They satisfy the required key points: high accuracy (a maximum mean squared error of 7.1x10^-5 for the unity-based normalized waveforms), efficiency (fully affordable computational effort for metamodel build-up: maximum 18 minutes on a general purpose computer), and simplicity (less than 1 second for running the metamodel, the user only provides the parameters combination). The metamodels can be used for very efficient generation of new waveforms, for any possible combination of dependent parameters, offering the possibility to explore the entire design space. A wide range of possibilities becomes achievable for the user, such as: all design corners can be analyzed, possible worst-case situations can be investigated, extreme values of waveforms can be discovered, sensitivity analyses can be performed (the influence of each parameter on the output waveform).     

EPG waveform characteristics of solenopsis mealybug stylet penetration on cotton

The solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), is a polyphagous insect known to cause severe damage to cotton (especially transgenic varieties) in South Asia, and currently poses a serious threat in Asia and potentially elsewhere. Stylet penetration behavior of P. solenopsis on cotton was monitored using the electrical penetration graph (EPG) technique (DC system) and the EPG characteristics were compared with those previously published from Phenacoccus manihoti Matile‐Ferrero and Planococcus citri (Risso). We identified and further characterized typical waveforms of A, B, C, and pd (together pathway), E1 and E2 (phloem), F (derailed stylet mechanics), and G (xylem). Five novel EPG aspects were distinguished in the EPG waveforms from P. solenopsis: (1) obvious B waveforms occurred following waveform A, (2) during waveform C, some aphid‐like E1e waveforms were observed, (3) prolonged potential drops (pd) up to >1 h occurred with two continuously alternating sub‐phases pd1 and pd2, (4) the pd1 waveform always occurred as the first waveform related to phloem sieve elements, preceding the other phloem waveforms (E), the labeling of which we changed to achieve a better comparison to the aphid E waveforms, and (5) waveform F, related to derailed stylet mechanics occurred but was not reported from other mealybugs so far. This is mainly a waveform morphology study to extend existing knowledge on mealybug EPGs to investigate mealybug‐host plant interactions. Further experimental verification of waveform correlations with plant tissue positions of stylet tips and insect activities is still needed.     

The effect of inlet waveforms on computational hemodynamics of patient-specific intracranial aneurysms

Due to the lack of patient-specific inlet flow waveform measurements, most computational fluid dynamics (CFD) simulations of intracranial aneurysms usually employ waveforms that are not patient-specific as inlet boundary conditions for the computational model. The current study examined how this assumption affects the predicted hemodynamics in patient-specific aneurysm geometries. We examined wall shear stress (WSS) and oscillatory shear index (OSI), the two most widely studied hemodynamic quantities that have been shown to predict aneurysm rupture, as well as maximal WSS (MWSS), energy loss (EL) and pressure loss coefficient (PLc). Sixteen pulsatile CFD simulations were carried out on four typical saccular aneurysms using 4 different waveforms and an identical inflow rate as inlet boundary conditions. Our results demonstrated that under the same mean inflow rate, different waveforms produced almost identical WSS distributions and WSS magnitudes, similar OSI distributions but drastically different OSI magnitudes. The OSI magnitude is correlated with the pulsatility index of the waveform. Furthermore, there is a linear relationship between aneurysm-averaged OSI values calculated from one waveform and those calculated from another waveform. In addition, different waveforms produced similar MWSS, EL and PLc in each aneurysm. In conclusion, inlet waveform has minimal effects on WSS, OSI distribution, MWSS, EL and PLc and a strong effect on OSI magnitude, but aneurysm-averaged OSI from different waveforms has a strong linear correlation with each other across different aneurysms, indicating that for the same aneurysm cohort, different waveforms can consistently stratify (rank) OSI of aneurysms.     

Electric organ discharge variability of Mormyridae (Teleostei: Osteoglossomorpha) in the Upper Volta system

The electric organ discharges (EODs) of five mormyrid species ( Marcusenius senegalensis , Brevimyrus niger , Petrocephalus bovei , Pollimyrus isidori , Hippopotamyrus pictus ) from different sampling sites from the Upper Volta system in West Africa were investigated. EOD waveforms were recorded at high sampling rates in order to compare signal waveform parameters of the different species from different locations. Except for H. pictus , EODs within a species differed significantly from one another in some parameters and waveform variability at least between some sampling sites. In addition, each species showed a continuous spectrum of waveform variations, all or only parts of which were found at certain localities. Although there was variability and sometimes similarities between species, the EOD waveforms were species specific. Knowing their variation spectrum, they can be used for species determination and are probably used for species recognition by the mormyrids. Similarities or differences in EOD waveform expression within a species were not related to geographical distance. By contrast, we suggest that biotic environmental factors at a given location influence the expression of EOD waveforms. These factors affect absolute measurements such as EOD duration and fast Fourier transformation peak frequency as well as the amount of variation for certain waveform parameters across species in a similar manner for a given site. Although EOD waveform might be important for the establishment of reproductive barriers between species, our results suggest that differences in waveforms may not necessarily reflect different species or speciation processes in progress.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 61–80.     

Parameter estimation in cardiac ionic models

We examine the problem of parameter estimation in mathematical models of excitable cell cardiac electrical activity using the well-known Beeler–Reuter (1977) ionic equations for the ventricular action potential. The estimation problem can be regarded as equivalent to the accurate reconstruction of ionic current kinetics and amplitudes in an excitable cell model, given only action potential experimental data. We show that in the Beeler–Reuter case, all ionic currents may be reasonably reconstructed using an experimental design consisting of action potential recordings perturbed by pseudo-random injection currents.

The Beeler–Reuter model was parameterised into 63 parameters completely defining all membrane current amplitudes and kinetics. Total membrane current was fitted to model-generated experimental data using a ‘data-clamp’ protocol. The experimental data consisted of a default action-potential waveform and an optional series of perturbed waveforms generated by current injections. Local parameter identifiability was ascertained from the reciprocal condition value (1/λ) of the Hessian at the known solution. When fitting to a single action potential waveform, the model was found to be over-determined, having a 1/λ value of 3.6e−14. This value improved slightly to 1.4e−10 when an additional 2 perturbed waveforms were included in the fitting process, suggesting that the additional data did not overly improve the identifiability problem. The additional data, however, did allow the accurate reconstruction of all ionic currents. This indicates that by appropriate experimental design, it may be possible to infer the properties of underlying membrane currents from observation of transmembrane potential waveforms perturbed by pseudo-random currents.     

Effect of waveforms of inspired gas tension on the respiratory oscillations of carotid body discharge

The responses of carotid body chemoreceptor discharge to repeated ramps (20- to 60-s forcing cycle durations) of inspired gas tensions were studied in spontaneously breathing and in artificially ventilated pentobarbitone-anesthetized cats. In all animals the mean intensity of chemoreceptor discharge followed the frequency of the forcing cycle, and superimposed on this were oscillations at the frequency of ventilation (breath-by-breath oscillations). The amplitude of the breath-by-breath oscillations in discharge was often large, and it waxed and waned with the forcing cycle. It was greatest when the mean level of discharge was falling and smallest near the peak of mean discharge. No qualitative differences were observed between PO2-alone forcing in constant normocapnia and PCO2-alone forcing in constant hypoxia. The variation in the amplitudes of breath-by-breath oscillations was shown to be due primarily to variations in the amplitudes of the downslope component of the discharge oscillation. Variations in the upslope component of individual oscillations were small. The factors responsible for the breath-by-breath oscillations are discussed, and it is concluded that the shape of the waveform of arterial gas tensions that stimulate the peripheral chemoreceptors departs markedly from that of a line joining end-tidal gas tensions. This causes breath-by-breath oscillations of discharge to be very large after an "off" stimulus. Reflex studies involving the forcing of respiratory gases should therefore include consideration of these effects.     

Determination of the average shape of flagellar bends: a gradient curvature model

Data obtained by manual digitization of photographs of flagellar bending waves have been analyzed by determining size parameters for the bends by least-squares fitting of a model waveform. These parameters were then used to normalize the data so that the average shape of the bends could be determined. Best fits were obtained with a model waveform derived from the constant curvature waveforms used previously but with provision for a linear change in curvature across the central region of the bend-the gradient curvature model (GCM). The central regions of the GCM bending waves are separated by transition regions with length determined by a parameter called the truncation factor (FT). Fitting the GCM to sine-generated bending waves give optimal fit when FT = 0.34. Fitting the GCM to four different samples of flagellar bending waves gave best fits with values of FT ranging from 0.17 for ATP-reactivated Lytechinus spermatozoa beating at approximately 10 Hz to 0.32 for live spermatozoa of Arbacia. The difference between the Arbacia waveforms and a sine-generated waveform is therefore very small, but a sine-generated waveform lacks the degree of freedom represented by FT that is required to fit other waveforms optimally. The residual differences between the waveform data and optimal GCM waveforms were averaged and found to be small. In most cases, the curvature in the central region of the optimal GCM decreased in magnitude towards the tip of the flagellum; however, this slope was highly variable and sometimes positive. Significant variations in both this slope and FT were found in individual bends as they propagated along a flagellum.     

Waveform characterization of the soybean stem feeder Edessa meditabunda: overcoming the challenge of wiring pentatomids for EPG

Stink bugs (Hemiptera: Heteroptera: Pentatomidae) are in general robust and restless insects, which makes them difficult to wire for electropenetrograph (EPG) studies. In addition, cuticular lipids may reduce wire effectiveness, and their removal could improve success of wiring. We compared wiring effectiveness for three species of stink bugs, differing in walking behaviour and degree of cuticular waxiness, that is, Piezodorus guildinii (Westwood), Nezara viridula (L.), and Loxa deducta (Walker). Results indicated that removal of cuticular lipids by mechanical abrasion (via sanding) greatly improved attachment success with gold wire. Our hypothesis that heavier and bigger bugs would lose the wire attachment more quickly than lighter and smaller bugs was not confirmed, regardless of the sanding. In contrast, our hypothesis that greater movement of a bug would cause the wire to break more often was supported by extensive testing. Behaviour appears to be more relevant for successful wiring than body weight. We used the sanding and wiring technique to characterize and correlate direct current EPG waveforms for the large and restless stem‐feeding stink bug Edessa meditabunda (Fabricius) on soybean plants. This marks the first published example of pentatomid EPG waveforms. Edessa meditabunda recordings on soybean stems generated eight types of waveforms in three phases and two families, named as follows: non‐probing = Np and Z; pathway phase = Em1; X wave phase = X; ingestion phase, family I = Em2 and Em3; ingestion phase, family N = Em4 and Em5. These eight were described based on their frequencies, relative amplitudes, and level voltages. Histological studies of stylets within salivary sheaths correlated the Em1, Em2, and Em3 waveforms with specific penetration sites. The waveform with the longest duration when feeding was Em2, representing xylem sap ingestion; in addition, waveform Em3 (always preceded by an X wave) was correlated with phloem sap ingestion.     

Strain waveform dependence of stress fiber reorientation in cyclically stretched osteoblastic cells: effects of viscoelastic compression of stress fibers

Actin stress fibers (SFs) of cells cultured on cyclically stretched substrate tend to reorient in the direction in which a normal strain of substrate becomes zero. However, little is known about the mechanism of this reorientation. Here we investigated the effects of cyclic stretch waveform on SF reorientation in osteoblastic cells. Cells adhering to silicone membranes were subjected to cyclic uniaxial stretch, having one of the following waveforms with an amplitude of 8% for 24 h: triangular, trapezoid, bottom hold, or peak hold. SF reorientation of these cells was then analyzed. No preferential orientation was observed for the triangular and the peak-hold waveforms, whereas SFs aligned mostly in the direction with zero normal strain (~55°) with other waveforms, especially the trapezoid waveform, which had a hold time both at loaded and unloaded states. Viscoelastic properties of SFs were estimated in a quasi-in situ stress relaxation test using intact and SF-disrupted cells that maintained their shape on the substrate. The dynamics of tension F(SFs) acting on SFs during cyclic stretching were simulated using these properties. The simulation demonstrated that F(SFs) decreased gradually during cyclic stretching and exhibited a compressive value (F(SFs) < 0). The magnitude and duration time of the compressive forces were relatively larger in the group with a trapezoid waveform. The frequency of SF orientation had a significant negative correlation with the applied compressive forces integrated with time in a strain cycle, and the integrated value was largest with the trapezoid waveform. These results may indicate that the applied compressive forces on SFs have a significant effect on the stretch-induced reorientation of SFs, and that SFs realigned to avoid their compression. Stress relaxation of SFs might be facilitated during the holding period in the trapezoid waveform, and depolymerization and reorientation of SFs were significantly accelerated by their viscoelastic compression.     

On-line analysis of neuromuscular bioelectric potentials

A computer system is presented which provides for on-line data capture and analysis of evoked end-plate potentials and action potentials, and on-line data capture with off-line analysis of spontaneously occurring miniature end-plate potentials at the end-plate region of the neuromuscular junction. Sampling of evoked waveforms begins after an adjustable delay following the stimulus. Spontaneously occurring waveforms are captured by 'freezing' the contents of a circular buffer. The software provides MENU selectable support functions including storage and retrieval of data and calculated parameters, analog and digital display of waveforms, data calibration and gain modification, data editing, file management, and hardcopy output. Calculated parameters of the waveform are optionally placed in a data base file by the analysis programs. The data base may be used for editing, arithmetic operations, and subsetting of variables as well as statistical analysis and plotting of any selected variables.     

Characterisation of the feeding behaviour of western flower thrips in terms of electrical penetration graph (EPG) waveforms

Western flower thrips, Frankliniella occidentalis (Pergande) causes damage to plants when they are feeding. Also, this thrips species transmits Tomato spotted wilt virus (TSWV) during stylet penetration. We investigated the penetration behaviour (probing) of thrips on pepper leaves and on liquid diet by electrical penetration graph (EPG, DC-system) recording. In addition, we used high-magnification video observations to correlate EPG waveforms with the insect's posture, head movements, and muscle contractions. Also, EPGs were correlated with probing on liquid diets containing radio-active tracers to distinguish and quantify ingestion waveforms. The previously described waveforms P, Q, and R were distinguished and additionally, a new waveform 'S' was distinguished. Waveform P could be linked with mandibular leaf penetration, waveform Q presumably with insertion of the maxillary stylets, and waveform R with ingestion of cell contents, whereas waveform S could not be correlated with any behavioural activity. Histology of the feeding damage in pepper leaves shows that thrips ingests the contents of multiple cells per probe.     

Trade-off between foliage and tuber resistance to Phthorimaea operculella in wild potatoes

Beet leafhopper, Circulifer tenellus (Baker) (Homoptera: Cicadellidae), is the only known North American vector of beet curly top virus (Geminiviridae), which causes major economic losses in a number of crops including sugar beet, tomato, beans, and peppers. Beet curly top virus is a phloem-limited, persistently transmitted, circulative geminivirus. The strain/species of curly top virus used in this study is the CFH strain, also referred to as beet severe curly top virus (BSCTV). The direct current (DC) electrical penetration graph technique was used to determine the specific stylet penetration behavior associated with inoculation of BSCTV. Viruliferous leafhoppers were allowed to feed on healthy 3–4-week-old sugar beet plants until specific electrical penetration graph waveforms were produced, at which point feeding was artificially terminated. A series of comparisons between leafhoppers that produced different combinations of waveforms clearly implicated waveform D1 as the only waveform correlated with inoculation of BSCTV. All successful inoculations contained waveform D1, and 56 out of 64 leafhoppers that produced waveform D1 successfully inoculated test plants. Eighty-five leafhoppers did not produce waveform D1 and none of these inoculated BSCTV. While the occurrence of waveform D1 appears to be necessary for BSCTV inoculation, there was no correlation between duration of waveform D1 and inoculation success rate. The correlation of waveform D1 and BSCTV inoculation found in this study implies that waveform D1 is associated with phloem salivation.     

园艺矿物油乳剂对亚洲柑橘木虱成虫刺吸行为的影响

目的】亚洲柑橘木虱Diaphorina citri是传播柑橘最重要病害黄龙病(huanglongbing, Candidatus Liberibacter asiaticus)的媒介昆虫,树冠喷施园艺矿物油(horticultural mineral oil, HMO)可以减少其在柑橘上的取食和产卵。本研究旨在探索植物组织内残留的园艺矿物油如何影响亚洲柑橘木虱成虫的取食行为。【方法】采用直流型刺吸电位仪(DC-EPG Giga-8)记录亚洲柑橘木虱成虫12 h内在喷施不同浓度(0.25%, 0.5%, 1%和2%, v/v)HMO(nC24)乳剂和清水(对照)时柑橘嫩叶上的刺探和取食行为并转换为波形信号,分析比较各处理的波形参数。【结果】不同浓度HMO乳剂处理显著提高了亚洲柑橘木虱成虫在柑橘叶片上的C波(路径波)持续时间百分比,显著减少了E2波(韧皮部吸食汁液)持续时间百分比,而对D(口针第一次接触韧皮部组织), E1(韧皮部出现唾液分泌) 和G(木质部吸食汁液) 波持续时间百分比没有显著影响。HMO处理组C波的每成虫波形累积时间(WDI)显著长于对照组,而各浓度处理之间没有显著差异,但D波WDI明显短于对照组。随着HMO浓度的增加, E1, E2和G波WDI显著下降,2% HMO处理组E1, E2和G波WDI明显低于其他浓度处理组。HMO乳剂处理明显减少了刺探次数;在HMO处理中,每成虫刺探次数和每成虫各波形次数均显著少于对照组,其中以1%和2% HMO乳剂处理最少。HMO乳剂处理组np波 (非刺探波)、D波、E2波的每成虫单次波形持续时间(WDE)均短于对照组,其中2% HMO乳剂处理后,D, E1和E2波WDE显著短于其他浓度处理和对照。同时,2% HMO乳剂处理推迟了从口器接触叶片到第1次开始刺探的时间和从口器接触叶片到第1次刺探到韧皮部的时间。【结论】结果表明,2% HMO乳剂显著减少亚洲柑橘木虱成虫在柑橘叶片上的取食次数和有效取食时间,同时还缩短了其在韧皮部分泌唾液和吸食汁液时间,因此可推荐矿物油用于柑橘黄龙病和亚洲柑橘木虱的综合防治体系。同时初步推断矿物油的作用机制可能是其进入植物气孔阻止植物挥发性物质的释放,抑制或掩盖柑橘叶片表面的挥发性物质从而减少了亚洲柑橘木虱在寄主上取食。     

Optimal temperature policy for immobilized enzyme packed bed reactor performing reversible Michaelis-Menten kinetics using the disjoint policy.

The optimal temperature policy that maximizes the time-averaged productivity of a continuous immobilized enzyme packed bed reactor is determined. This optimization study takes into consideration the enzyme thermal deactivation with substrate protection during the reactor operation. The general case of reversible Michaelis-Menten kinetics under constant reactor feed flow rate is assumed. The corresponding nonlinear optimization problem is solved using the calculus of variations by applying the disjoint policy. This policy reduces the optimization problem into a differential-algebraic system, DAE. This DAE system defines completely the optimal temperature-time profiles. These profiles depend on the kinetic parameters, feed substrate concentration, operating period, and the residence time and are characterized by increasing form with time. Also, general analytical expressions for the slopes of the temperature and residual enzyme activity profiles are derived. An efficient solution algorithm is developed to solve the DAE system, which results into a one-dimensional optimization problem with simple bounds on the initial feed temperature. The enzymatic isomerization of glucose into fructose is selected as a case study. The computed productivities are very close to that obtained by numerical nonlinear optimization with simpler problem to solve. Moreover, the computed conversion profiles are almost constant over 90% of the operating periods, thus producing a homogeneous product.     

Characterization of the electrical penetration graphs of the psyllid Bactericera trigonica on carrots

The psyllid Bactericera trigonica Hodkinson (Hemiptera: Triozidae) is a carrot and celery pest recently described as a vector of the plant pathogenic bacterium Candidatus Liberibacter solanacearum (Lso) on Apiaceae. Detailed information on vector stylet penetration activities is essential in the study of Lso transmission. In this study we used the electrical penetration graph (EPG) technique, characterized waveforms produced during the various stylet penetration activities in carrot leaves, and correlated them with stylet tracks and salivary sheath termini on plant tissues as well as with Lso inoculation. In addition, the effect of Lso in B. trigonica on the stylet penetration activities was tested. The EPG waveforms identified were: waveforms C1 and C2 detected in the mesophyll, waveforms D, E1, and E2 near or in the phloem sieve elements, and waveform G in the xylem vessels. A waveform pattern not previously reported for psyllids was the ‘pseudo‐potential drop’ (pseudo‐pd), characterized by sudden voltage dips similar to potential drops. However, the lowered voltage appeared to be inverted when the plant voltage is negative, indicating that it is caused by an increased resistance period and not due to a cell puncture. A direct correlation is shown between the waveform E1 and salivation into phloem sieve elements by B. trigonica as the inoculation of Lso occurred in a period as short as 30 s of E1; Lso transmission occurred in 17 of 35 plants (48%). Stylet activities during waveforms C or D had no consequences on the inoculation of Lso. In conclusion, Lso infection directly affects the probing behaviour of B. trigonica by increasing the total duration of C and D waveforms, but not variables related to phloem salivation (Lso inoculation) or ingestion (Lso acquisition). The reported information here is fundamental for identifying the psyllid vector traits of behaviour associated with transmission of Lso to Apiaceae.     

Blind identification of the aortic pressure waveform from multiple peripheral artery pressure waveforms

We have developed a new technique to estimate the clinically relevant aortic pressure waveform from multiple, less invasively measured peripheral artery pressure waveforms. The technique is based on multichannel blind system identification in which two or more measured outputs (peripheral artery pressure waveforms) of a single-input, multi-output system (arterial tree) are mathematically analyzed so as to reconstruct the common unobserved input (aortic pressure waveform) to within an arbitrary scale factor. The technique then invokes Poiseuille's law to calibrate the reconstructed waveform to absolute pressure. Consequently, in contrast to previous related efforts, the technique does not utilize a generalized transfer function or any training data and is therefore entirely patient and time specific. To demonstrate proof of concept, we have evaluated the technique with respect to four swine in which peripheral artery pressure waveforms from the femoral and radial arteries and a reference aortic pressure waveform from the descending thoracic aorta were simultaneously measured during diverse hemodynamic interventions. We report that the technique reliably estimated the entire aortic pressure waveform with an overall root mean squared error (RMSE) of 4.6 mmHg. For comparison, the average overall RMSE between the peripheral artery pressure and reference aortic pressure waveforms was 8.6 mmHg. Thus the technique reduced the RMSE by 47%. As a result, the technique also provided similar improvements in the estimation of systolic pressure, pulse pressure, and the ejection interval. With further successful testing, the technique may ultimately be employed for more precise monitoring and titration of therapy in, for example, critically ill and hypertension patients.     

Characterization of electrical penetration graphs of the Asian citrus psyllid, Diaphorina citri, in sweet orange seedlings

Detailed information on probing behavior of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is critical for understanding the transmission process of phloem‐limited bacteria (Candidatus Liberibacter spp.) associated with citrus ‘huanglongbing’ by this vector. In this study, we investigated stylet penetration activities of D. citri on seedlings of Citrus sinensis (L.) Osbeck cv. Pêra (Rutaceae) by using the electrical penetration graph (EPG‐DC system) technique. EPG waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration into plant tissues. The main waveforms were correlated with histological observations of salivary sheath termini in plant tissues, to determine the putative location of stylet tips. The behavioral activities were also inferred based on waveform similarities in relation to other Sternorrhyncha, particularly aphids and whiteflies. In addition, we correlated the occurrence of specific waveforms with the acquisition of the phloem‐limited bacterium Ca. Liberibacter asiaticus by D. citri. The occurrence of a G‐like xylem sap ingestion waveform in starved and unstarved psyllids was also compared. By analyzing 8‐h EPGs of adult females, five waveforms were described: (C) salivary sheath secretion and other stylet pathway activities; (D) first contact with phloem (distinct from other waveforms reported for Sternorrhyncha); (E1) putative salivation in phloem sieve tubes; (E2) phloem sap ingestion; and (G) probably xylem sap ingestion. Diaphorina citri initiates a probe with stylet pathway through epidermis and parenchyma (C). Interestingly, no potential drops were observed during the stylet pathway phase, as are usually recorded in aphids and other Sternorrhyncha. Once in C, D. citri shows a higher propensity to return to non‐probing than to start a phloem or xylem phase. Several probes are usually observed before the phloem phase; waveform D is observed upon phloem contact, always immediately followed by E1. After E1, D. citri either returns to pathway activity (C) or starts phloem sap ingestion, which was the longest activity observed.     

Preparative purification of a recombinant protein by hydrophobic interaction chromatography: modulation of selectivity by the use of chaotropic additives

Development and implementation of a chaotropic wash step following protein loading on a hydrophobic interaction chromatographic (HIC) column is described for the purification of a recombinant protein. Various agents that reduce protein affinity in hydrophobic interaction chromatographic systems were screened for their utility in a wash step following protein loading on a Phenyl Fast Flow Sepharose HIC column. A combination of sodium thiocyanate, glycerol, and urea was selected as a suitable additive for the wash buffer that selectively eluted most of the major impurities present in the feed stream. Eluate purity, as monitored by reversed-phase chromatography and SDS-PAGE, was significantly increased by incorporation of this wash step in the purification process. Incorporation of this wash step on HIC enabled a reduction in the overall number of chromatographic steps in the downstream purification process for this recombinant protein, resulting in improved process yields and significant economic advantages.The effect of varying concentrations of each of the three wash additives on yield was studied. While the step yield decreased with an increase in concentration for urea and sodium thiocyanate, an optimum was observed with respect to glycerol concentration. The preferential interaction theory is employed to explain this effect.