Damage Evolution of Ballastless Track Concrete Exposed to Flexural Fatigue Loads:The Application of Ultrasonic Pulse Velocity,Impact-echo and Surface Electrical Resistance Method

In order to clarify the fatigue damage evolution of concrete exposed to flexural fatigue loads,ultrasonic pulse velocity(UPV),impact-echo technology and surface electrical resistance(SR) method were used.Damage variable based on the change of velocity of ultrasonic pulse(Du) and impact elastic wave(Di)were defined according to the classical damage theory.The influences of stress level,loading frequency and concrete strength on damage variable were measured.The experimental results show that Du and Di both present a three-stages trend for concrete exposed to fatigue loads.Since impact elastic wave is more sensitive to the microstructure damage in stage Ⅲ,the critical damage variable,i e,the damage variable before the final fracture of concrete of Di is slightly higher than that of Du.Meanwhile,the evolution of SR of concrete exposed to fatigue loads were analyzed and the relationship between SR and Du,SR and Di of concrete exposed to fatigue loads were established.It is found that the SR of concrete was decreased with the increasing fatigue cycles,indicating that surface electrical resistance method can also be applied to describe the damage of ballastless track concrete exposed to fatigue loads.

Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing

BACKGROUND Mesenchymal stem cells(MSCs)modulated by various exogenous signals have been applied extensively in regenerative medicine research.Notably,nanosecond pulsed electric fields(nsPEFs),characterized by short duration and high strength,significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways.Consequently,we used transcriptomics to study changes in messenger RNA(mRNA),long noncoding RNA(lncRNA),microRNA(miRNA),and circular RNA expression during nsPEFs application.AIM To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs.METHODS The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing.MSCs were pretreated with 5-pulse nsPEFs(100 ns at 10 kV/cm,1 Hz),followed by total RNA isolation.Each transcript was normalized by fragments per kilobase per million.Fold change and difference significance were applied to screen the differentially expressed genes(DEGs).Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions,complemented by quantitative polymerase chain reaction verification.RESULTS In total,263 DEGs were discovered,with 92 upregulated and 171 downregulated.DEGs were predominantly enriched in epithelial cell proliferation,osteoblast differentiation,mesenchymal cell differentiation,nuclear division,and wound healing.Regarding cellular components,DEGs are primarily involved in condensed chromosome,chromosomal region,actin cytoskeleton,and kinetochore.From aspect of molecular functions,DEGs are mainly involved in glycosaminoglycan binding,integrin binding,nuclear steroid receptor activity,cytoskeletal motor activity,and steroid binding.Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation.CONCLUSION Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs,2 miRNAs,and 65 lncRNAs.Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways,which are involved in vesicular transport,calcium ion transport,cytoskeleton,and cell differentiation.

Application of a Pulsed Electric Field to Modify the Free Energy of the Active Site of an Azoreductase

The Gibbs free energy is strongly related to the stability and catalytic function of an enzyme through the energetic changes that occur in the chemical reactions the enzyme catalyzes. In this in silico study, a pulsed electric field was applied to an azoreductase, and its effect on the Gibbs free energy of molecular docking with two dyes was measured. We propose that certain stimuli from a pulsed electric field favor the structural stability of the enzyme by promoting an arrangement in the active site, potentially leading to an enhancement of enzymatic activity overall.

Effect of low-frequency pulse percutaneous electric stimulation on peripheral nerve injuries at different sites

BACKGROUND： The postoperative recovery of nerve function in patients with peripheral nerve injury is always an important problem to solve after treatment. The electric stimulation induced electromagnetic field can nourish nerve, postpone muscular atrophy, and help the postoperative neuromuscular function. OBJECTIVE： To observe the effects of low-frequency pulse percutaneous electric stimulation on the functional recovery of postoperative patients with peripheral nerve injury, and quantitatively evaluate the results of electromyogram （EMG） examination before and after treatment. DESIGN ： A retrospective case analysis SETTING： The Sixth People＇s Hospital affiliated to Shanghai Jiaotong University PARTICIPANTS： Nineteen postoperative inpatients with peripheral nerve injury were selected from the De- partment of Orthopaedics, the Sixth People＇s Hospital affiliated to Shanghai Jiaotong University from June 2005 to January 2006, including 13 males and 6 females aged 24-62 years with an average of 36 years old. There were 3 cases of brachial plexus nerve injury, 3 of median nerve injury, 7 of radial nerve injury, 3 of ul- nar nerve injury and 3 of common peroneal nerve injury, and all the patients received probing nerve fiber restoration. Their main preoperative manifestations were dennervation, pain in limbs, motor and sensory disturbances. All the 19 patients were informed with the therapeutic program and items for evaluation. METHODS： ① Low-frequency pulse percutaneous electric stimulation apparatus： The patients were given electric stimulation with the TERESA cantata instrument （TERESA-0, Shanghai Teresa Health Technology, Co., Ltd.）. The patients were stimulated with symmetric square waves of 1-111 Hz, and the intensity was 1.2-5.0 mA, and it was gradually adjusted according to the recovered conditions of neural regeneration following the principle that the intensity was strong enough and the patients felt no obvious upset. They were treated for 4- 24 weeks, 10-30 minutes for each time, 1-3 times a day, and 6 weeks as a course. ② EMG examination was applied to evaluate the recoveries of recruitment, motor conduction velocity （MCV） and sensory conduction velocity （SCV） before and after treatment. The patients were examined with the EMG apparatus （DIS- A2000C, Danmark） before and after the treatment of percutaneous electric stimulation. ③Standards for evaluating the effects included cured （complete recovery of motor functions, muscle strength of grade 5, no abnormality in EMG examination）, obviously effective [general recovery of motor function, muscle strength of grade 4, no or a few denervation potentials, motor conduction velocity （MCV） and sensory conduction velocity （SCV）], improved （partial recovery of motor function, muscle strength of grade 3, denervation potentials and reinneration potentials, slowed MCV and SCV, invalid （no obvious changes of motor function）. MAIN OUTCOME MEASURES： ① Ameliorated degree of the nerve function of the postoperative patients with peripheral nerve injury treated with percutaneous electric stimulation; ② Changes of EMG examination before and after treatment. RESULTS： All the 19 postoperative patients with peripheral nerve injury were involved in the analysis of results. ① Comparison of nerve function before and after treatment in 19 patients with peripheral nerve injury of different sites： For the patients with radial nerve injury （n=7）, the nerve functions all completely recovered after 8-week treatment, and the cured and obvious rate was 100% （7/7）; For the patients with brachial plexus nerve injury （n=3）, 1 case had no obvious improvement, and the cured and obvious rate was 67% （2/3）; For the patients with common peroneal nerve injury （n=3）, the extension of foot dorsum generally recovered in 1 case of nerve contusion after 4-week treatment, and the cured and obvious rate was 67% （2/3）; For the patients with median nerve injury （n=3）, muscle strength was obviously recovered, and the cured and obvious rate was 100% （3/3）; For the patients with ulnar nerve injury （n=3）, 1 case only had recovery of partial senses, and the cured and obvious rate was 67% （2/3）. Totally 9 cases were cured, 7 were obviously effective, 1 was improved, and only 2 were invalid. After 4 courses, the cured rate of damaged nerve function after four courses was 47% （9/19）, and effective rate was 89% （17/19）.② Comparison of EMG examination before and after treatment： Before and after percutaneous electric stimulation, he effective rates of recruitment, MCV and SCV were 89% （17/19）, 58% （11/19）, 47% （9/19） respectively, and there were extremely obvious differences （P〈 0.01）. CONCLUSION： ①Low-frequency pulse percutaneous electric stimulation can improve the nerve function of postoperative patients with peripheral nerve injury of different sites, especially that the injuries of radial nerve and median nerve recover more obviously. ②Percutaneous electric stimulation can ameliorate the indexes of EMG examination, especially the recruitment, in postoperative patients with peripheral nerve injury.

Treatment time influences the effects of a low-frequency pulsed electric field on synthesis of tyrosine hydroxylase and dopamine in PC12 cells

BACKGROUND： Electromagnetic radiation can influence dopamine （DA） synthesis in brain tissues or ceils, but electromagnetic frequencies, intensities, and radiation time can produce different effects. In addition, the signal pathway by which electromagnetic radiation influences DA synthesis remains controversial. OBJECTIVE： To determine tyrosine hydroxylase （TH） expression in PC12 cells and DA levels in cell culture media after different periods of low-frequency pulsed electric field （LF-PEF） stimulation, and to determine how LF-PEF signaling stimulates TH synthesis using inhibitors. DESIGN, TIME AND SETTING： A parallel, controlled, cell experiment was performed at the Laboratory of Cell Biology, School of Life Science, East China Normal University, between January and October 2006. MATERIALS： PC12 cells were purchased from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, China. Nerve growth factor was purchased from PeproTech, USA. The protein kinase A inhibitor, H-89, and mitogen-activated protein kinase kinase inhibitor, U0126, were purchased from Sigma, USA. METHODS： （1） Following routine culture in Dulbecco＇s modified eagle medium, primary PC12 cells were stimulated under LF-PEF （pulse frequency 50.Hz, pulse width 20 μs, peak field strength 1 V/m） for 5, 10, 15, 20, and 30 minutes. （2） Inhibitors （H-89 or U0126, 1 μmol/L） were added 30 minutes before LF-PEF stimulation for 10 minutes. MAIN OUTCOME MEASURES： （1） TH expression was determined by Western blot in PC12 cells at 0.5, 1,2, 3, and 4 days after LF-PEF stimulation. Similarly, DA was measured by high-performance liquid chromatography in media at 2, 3, 4, or 5 days after LF-PEE （2） TH expression was detected 1 day after H-89 or U0126 treatment and LF-PEE RESULTS： （1） Short-term LF-PEF stimulation （5 and 10 minutes） increased TH expression and media DA levels after short-term culture （2 days） （P 〈 0.01）, but both parameters decreased with longer culture （3 4 days） （P 〈 0.01）. Long-term LF-PEF stimulation （15, 20, or 30 minutes） decreased TH and DA synthesis, followed by a rapid increase （P 〈 0.01）. （2） H89 could completely inhibit TH expression in PC12 cells stimulated by LF-PEF for 10 minutes, while the inhibition rate of U0126 was 53.2%. CONCLUSION： Short-term LF-PEF first promotes then inhibits, while long-term LF-PEF first inhibits then promotes, TH and DA synthesis. LF-PEF stimulation regulates TH expression primarily by activating protein kinase A to regulate DA synthesis.

The influence of pore characteristics on rock fragmentation mechanism by high-voltage electric pulse

High-voltage electric pulse(HVEP)is an innovative low-energy and high-efficiency technique.However,the underlying physics of the electrical breakdown within the rock,and the coupling mechanism between the various physical fields involved in HVEP still need to be further understood.In this study,we establish a 2D numerical model of multi-physical field coupling of the electrical breakdown of porous rock with randomly distributed pores to investigate the effect of pore characteristics(porosity,pore media composition)on the partial electrical breakdown of rock(i.e.the generation of a plasma channel inside the rock).Our findings indicate that the generation of a plasma channel is directionally selective and extends in the direction of a weak electrical breakdown intensity.As the porosity of the rock increases,so does the intensity of the electric field in the‘electrical damage’region—the greater the porosity,the greater the effectiveness of rock-breaking.As the fraction of pore fluid(S_(water)/S_(air))gradually declines,the generation time of the plasma channel decreases,and the efficacy of rock-breaking by HVEP increases.In addition,in this study,we conducted an indoor experiment utilizing an electric pulse drill to break down the rock in order to recreate the growth mode of the plasma channel in the rock.Moreover,the experimental results are consistent with the simulation results.In addition,the development of this type of partial electrical breakdown is confirmed to be related to electrode polarity and pore characteristics via the experiment of the symmetrical needle-needle electrode arrangement,which further demonstrates the mechanism of partial electrical breakdown.This research is significant for comprehending the process of electric impulse rock-breaking and gives theoretical guidance and technological support for advancing electric impulse drilling technology.

Experimental and numerical investigation on the uniformity of nanosecond pulsed dielectric barrier discharge influenced by pulse parameters

Nanosecond(ns)pulsed dielectric barrier discharge(DBD)is considered as a promising method to produce controllable large-volume and high activity low-temperature plasma at atmospheric pressure,which makes it suitable for wide applications.In this work,the ns pulse power supply is used to excite Ar DBD and the influences of the pulse parameters(voltage amplitude,pulse width,pulse rise and fall times)on the DBD uniformity are investigated.The gas gap voltage(Ug)and conduct current(Ig)are separated from the measured voltage and current waveforms to analyze the influence of electrical parameters.The spectral line intensity ratio of two Ar excited species is used as an indicator of the electron temperature(Te).The time resolved discharge processes are recorded by an intensified charge-coupled device camera and a one-dimensional fluid model is employed to simulate the spatial and temporal distributions of electrons,ions,metastable argon atoms and Te.Combining the experimental and numerical results,the mechanism of the pulse parameters influencing on the discharge uniformity is discussed.It is shown that the space electric field intensity and the space particles'densities are mainly responsible for the variation of discharge uniformity.With the increase of voltage and pulse width,the electric field intensity and the density of space particles increased,which results in the discharge mode transition from non-uniform to uniform,and then non-uniform.Furthermore,the extension of pulse rise and fall times leads to the discharge transition from uniform to nonuniform.The results are helpful to reveal the mechanism of ns pulsed DBD mode transition and to realize controllable and uniform plasma sources at atmospheric pressure.

Demulsification Behavior, Characteristics, and Performance of Surfactant Stabilized Oil-in-Water Emulsion under Bidirectional Pulsed Electric Field

As a novel electric demulsification method,bidirectional pulsed electric field(BPEF)was employed to demulsify the surfactant stabilized oil-in-water(SSO/W)emulsion for oil/water separation in this work.The demulsification behavior,characteristics,and stages under BPEF were explored.It was discovered that BPEF drove SSO/W emulsion to move and form vortexes,during which the oil droplets aggregated and accumulated to generate an oil droplet layer(ODL).ODL subsequently transformed into a continuous oil layer(COL)leading to the demulsification and separation of SSO/W emulsion.The conversion rate of ODL to COL was defined and used to evaluate the demulsification process and reflect the coalescence ability and transformation efficiency of dispersed oil droplets into COL.Furthermore,the effects of BPEF voltage,frequency,duty cycle,ratio of pulse output time,and surfactant type and content on the demulsification performance were examined.The optimal values of BPEF parameters for demulsification operation were 400 V,25 Hz,50%,and 4:1.O/W emulsion containing anionic surfactant was apt to be demulsified by BPEF,nonionic surfactant took the second place and cationic surfactant was the most difficult.A high surfactant content was not conducive to the BPEF demulsification.This work is anticipated to provide useful guidance for oil/water separation and oil recovery from actual emulsified oily wastewater by BPEF.

Low-frequency pulsed electromagnetic field pretreated bone marrow-derived mesenchymal stem cells promote the regeneration of crush-injured rat mental nerve

Bone marrow-derived mesenchymal stem cells （BMSCs） have been shown to promote the regeneration of injured peripheral nerves. Pulsed electromagnetic field （PEMF） reportedly promotes the proliferation and neuronal differentiation of BMSCs. Low-frequency PEMF can induce the neuronal differentiation of BMSCs in the absence of nerve growth factors. This study was designed to investigate the effects of low-frequency PEMF pretreatment on the proliferation and function of BMSCs and the effects of low-frequency PEMF pre-treated BMSCs on the regeneration of injured peripheral nerve using in vitro and in vivo experiments.In in vitro experiments, quantitative DNA analysis was performed to determine the proliferation of BMSCs, and reverse transcription-polymerase chain reaction was performed to detect S100 （Schwann cell marker）, glial fibrillary acidic protein （astrocyte marker）, and brain-derived neurotrophic factor and nerve growth factor （neurotrophic factors） mRNA expression. In the in vivo experiments, rat models of crush-injured mental nerve established using clamp method were randomly injected with low-frequency PEMF pretreated BMSCs, unpretreated BMSCs or PBS at the injury site （1 × 106 cells）. DiI-labeled BMSCs injected at the injury site were counted under the fluorescence microscope to determine cell survival. One or two weeks after cell injection, functional recovery of the injured nerve was assessed using the sensory test with von Frey filaments. Two weeks after cell injection, axonal regeneration was evaluated using histomorphometric analysis and retrograde labeling of trigeminal ganglion neurons. In vitro experiment results revealed that low-frequency PEMF pretreated BMSCs proliferated faster and had greater mRNA expression of growth factors than unpretreated BMSCs. In vivo experiment results revealed that compared with injection of unpretreated BMSCs, injection of low-frequency PEMF pretreated BMSCs led to higher myelinated axon count and axon density and more DiI-labeled neurons in the trigeminal ganglia, contributing to rapider functional recovery of injured mental nerve. These findings suggest that low-frequency PEMF pretreatment is a promising approach to enhance the efficacy of cell therapy for peripheral nerve injury repair.

Heredity of Aluminum Melt Caused by Electric Pulse Modification (Ⅰ)

The heredity of aluminum melt under the action of pulse electric field was investigated by means of the remelt experiment. A new hereditary criterion under this condition was proposed; in the meantime, the differential transferability of genetic carrier in activated melt among filial generations was validated with the aid of DSC.

Removal of phenol by activated alumina bed in pulsed high-voltage electric field

A new process for removing the pollutants in aqueous solution-activated alumina bed in pulsed high-voltage electric field was investigated for the removal of phenol under different conditions. The experimental results indicated the increase in removal rate with increasing applied voltage, increasing pH value of the solution, aeration, and adding Fe^2＋. The removal rate of phenol could reach 72.1% when air aeration flow rate was 1200 ml/min, and 88.2% when 0.05 mmol/L Fe^2＋ was added into the solution under the conditions of applied voltage 25 kV, initial phenol concentration of 5 mg/L, and initial pH value 5.5. The addition of sodium carbonate reduced the phenol removal rate. In the pulsed high-voltage electric field, local discharge occurred at the surface of activated alumina, which promoted phenol degradation in the thin water film. At the same time, the space-time distribution of gas-liquid phases was more uniform and the contact areas of the activated species generated from the discharge and the pollutant molecules were much wider due to the effect of the activated alumina bed. The synthetical effects of the pulsed high-voltage electric field and the activated alumina particles accelerated phenol degradation.

Effect of electric pulse treatment on solidification structure of an Al-15%Si alloy

The effect of electric pulse modifying on the solidification structure of an Al-15%Si alloy was investigated. The result shows that the primary silicon disappears sometimes and the eutectic phase is refined after the treatrnent of EP （electric pulse） though there are different modalities in different treating durations. DSC （differential scanning calorimetry） analysis indicates that the super-cooling texture decreases and the freezing range narrows evidently after the electric pulse treatment.

Heredity of Aluminum Melt by Electric Pulse Modification (Ⅱ)

Heredity of high pure aluminum melts under different pulse electric field was investigated by means of repetitious remelt experiment. The results indicate that the genetic coefficient by measurement of grain size of cast structure has a close relation with pulse voltage. Moreover, the hereditary law accords with the function of In = 1＋ e^-an＋β. The stability of genetic carrier （cluster） comprises in the competition between repetitious cooling and heating impulse and the effect of electric pulse modification.

Recent progress in pulsed electric field ablation for liver cancer

The number of liver cancer patients is likely to continue to increase in the coming decades due to the aging of the population and changing risk factors.Traditional treatments cannot meet the needs of all patients.New treatment methods evolved from pulsed electric field ablation are expected to lead to breakthroughs in the treatment of liver cancer.This paper reviews the safety and efficacy of irreversible electroporation in clinical studies,the methods to detect and evaluate its ablation effect,the improvements in equipment and its antitumor effect,and animal and clinical trials on electrochemotherapy.We also summarize studies on the most novel nanosecond pulsed electric field ablation techniques in vitro and in vivo.These research results are certain to promote the progress of pulsed electric field in the treatment of liver cancer.

Casting structure of pure aluminum by electric pulse modification at different superheated temperatures

Electric pulse modification （EPM） is a novel technique that reduces grain size by altering the structure of a melt. It was investigated that the response of the casting structure of high pure aluminum to EPM in different superheated melts. The results indicate that the grain refining effect of a given pulse electric field holds an optimal temperature range, moreover, a lower or higher superheated temperature will both disadvantage the improvements of casting structure. It essentially lies in the cooperative action between the distorted absorption of clusters and the activated capability of atoms in the aluminum melt.

Effects of electric pulse on microstructure of Al-Si alloy in liquid and solid states

In order to investigate the change in liquid microstructure of Al-Si alloytreated by electric pulse (EP), X-ray diffraction tests with liquid Al-Si alloy and ZL109 alloytreated or not by EP were carried out. The results show that the number of Al-Si atomic clustersdecreases and that of Al-Al and Si-Si atomic clusters increases for the treated samples. The testswith ZL109 alloy indicate that a large amount of primary crystal Si appears in the solidifiedmicrostructure after treated by EP. It is found that EP can change the microstructure of liquidmetal by affecting the probability of electrons appearing in different atoms (Al and Si) in theliquid metal. The combining force of different atoms decreases relatively, and that of the sameatoms increases, which is the main reason of reducing the atomic cluster with different atoms(Al-Si) and increasing the atomic cluster with the same atoms (Al-Al, Si-Si). The increasing of theatomic cluster with the same atom cluster resulted in the increasing of Si activity and the higherpoint of eutectics in the phase diagram. It makes a lot of primary silicon appeared in ZL109 alloy.

Effects of High-voltage Pulse Electric Field Treatment on the Structure Stability of Konjac Glucomannan

Structures of KGM treated in two high-voltage pulse electric fields were characterized by infrared spectroscopy,Raman spectroscopy,X-ray diffraction and so on.The results showed that intermolecular hydrogen bonding interactions of KGM were reduced after being treated with high-voltage pulse electric field,but there was no significant effect on its fiber chain form and thermal characteristics.Results of the study can provide a useful reference for further study on the structure and property of KGM,and especially can provide theoretical basis for the effect of physical field on the foodstuff deep processing related to KGM.

Microstructure and Temperature Distribution in ZnAl_2O_4 Sintered Body by Pulse Electric Current

Microstructure of reaction sintering of ZnAl2O4 at 1500℃ by hot-pressing(HP) and pulse electric current was investigated. The results indicated that the existed cracks in sintered body were caused by structure mismatch. It is the evidence that periodical temperature field existed during pulse electric current sintering of nonconductive materials. The distance between high temperature areas was related to die diameter.

Discharge Characteristics of SF_6 in a Non-Uniform Electric Field Under Repetitive Nanosecond Pulses

The characteristics of high pressure sulphur hexafluoride（SF6） discharges in a highly non-uniform electric field under repetitive nanosecond pulses are investigated in this paper.The influencing factors on discharge process,such as gas pressure,pulse repetition frequency（PRF）,and number of applied pulses,are analyzed.Experimental results show that the corona intensity weakens with the increase of gas pressure and strengthens with the increase of PRF or number of applied pulses.Spark discharge images suggest that a shorter and thicker discharge plasma channel will lead to a larger discharge current.The number of applied pulses to breakdown descends with the increase of PRF and ascends with the rise of gas pressure.The reduced electric field（E/p） decreases with the increase of PRF in all circumstances.The experimental results provide significant supplements to the dielectric characteristics of strongly electronegative gases under repetitive nanosecond pulses.

Lorentz force electrical impedance tomography using pulse compression technique

Lorentz force electrical impedance tomography （LFEIT） combines ultrasound stimulation and electromagnetic field detection with the goal of creating a high contrast and high resolution hybrid imaging modality. In this study, pulse compression working together with a linearly frequency modulated ultrasound pulse was investigated in LFEIT. Experiments were done on agar phantoms having the same level of electrical conductivity as soft biological tissues. The results showed that：（i） LFEIT using pulse compression could detect the location of the electrical conductivity variations precisely; （ii） LFEIT using pulse compression could get the same performance of detecting electrical conductivity variations as the traditional LFEIT using high voltage narrow pulse but reduce the peak stimulating power to the transducer by 25.5 dB; （iii） axial resolution of 1 mm could be obtained using modulation frequency bandwidth 2 MHz.