Uncoating of influenza virus in endosomes

The intracellular uncoating site of influenza virus was studied by measuring the fluorescence intensity of probes conjugated to the virus or the isolated hemagglutinin and also by assaying virus replication under various incubation conditions. Acidification of the viral environment was monitored by the decrease in the fluorescence intensity of fluorescein isothiocyanate, and transport of the virus particles into secondary lysosomes was assayed by the increase in the fluorescence intensity of fluorescein isothiocyanate diphosphate. The intracellular pH was estimated by the ratio of fluorescence intensities excited at two different wavelengths. It was found that the viral environment became acidified to a pH value of 5.1 to 5.2 within 10 min at 37 degrees C or 1 h at 20 degrees C after endocytosis. Addition of ammonium chloride to the medium rapidly raised the pH to 6.7. Transport of the virus particles into the secondary lysosomes was slower and negligibly low during those incubation periods. Virus replication occurred when the cells were incubated for 10 min at 37 degrees C or for 1 h at 20 degrees C, followed by incubation in the presence of ammonium chloride for a total of 12 h. These results indicate the uncoating of influenza virus in endosomes before reaching the secondary lysosomes.     

Participation of H+ in the Ca2(+)-induced conformational transition of 4-nitro-2,1,3-benzoxadiazole-labeled sarcoplasmic reticulum ATPase

The binding of Ca2+ to 4-nitro-2,1,3-benzoxadiazole (NBD)-labeled sarcoplasmic reticulum Ca2(+)-ATPase was accelerated markedly when the pH was changed at 11 degrees C from 6.5 to 8.0 at the time of Ca2+ addition. We examined the effect of pH on the enzyme conformational transition by measuring the kinetics of NBD fluorescence rises induced by a pH jump under various ligand conditions. The fast fluorescence rise following a pH jump from 6.0 or 6.5 to various test pHs in the presence and absence of Ca2+ proceeded monoexponentially. The amplitude of this fluorescence rise in the presence of Ca2+ was independent of the test pH, whereas the observed rate constant (kobs) increased markedly as the test pH increased. In contrast, the amplitude of the fast fluorescence rise in the absence of Ca2+ increased with increasing test pH, whereas kobs decreased. MgATP or Mg2+ influenced the pH dependences of these parameters in a complex way except for the amplitudes measured in the presence of Ca2+. These data could be simulated by using a reaction model in which Ca2+ binding is preceded by a rate-limiting enzyme conformational transition from a low to a high NBD fluorescence state and 1 mol each of H+ is liberated before and after this conformational transition. MgATP or Mg2+ appeared to promote this conformational transition by enhancing deprotonation of the enzyme. These results suggest that deprotonation may be the primary event in the activation of the unphosphorylated enzyme by Ca2+.     

Use of Tunable, Pulsed Dye Laser for Quantitative Fluorescence in Syphilis Serology (FTA-ABS Test)

A pulsed dye laser was used as an excitation source in a fluorescent treponemal antibody absorption (FTA-ABS) test. A high precision in quantitative fluorescence was obtained with this high-power excitation source coupled to an electronic detection system and a storage oscilloscope by standardization of fluorescence evaluation and through elimination of human error. One 0.4-mus pulse exposure was sufficient to record fluorescence intensity data on the oscilloscope. Absence of fading of fluorescence after repeated excitation permitted multiple readings of the same microscope field. Almost 100% reproducible results were obtained for the FTA-ABS test with 40 samples. Electronic detection of fluorescence and the high sensitivity obtained with laser excitation raise doubts about the relative value of quantitative immunofluorescence in the FTA-ABS test.     

Standardization of the conditions for measuring the specific fluorescence intensity in continuous cell cultures infected with variants of the Japanese encephalitis virus

The use of modernized photometric accessories to a model microscope manufactured by the Leningrad Optico-Mechanical Amalgamation (USSR), as well as the practical application of innovations facilitating and standardizing research work, has made it possible to obtain objective data indicating the presence of significant, direct, linear correlation between infectious activity and the intensity of fluorescence emitted by fluorescent antibodies bound with the antigen of 11 studied variants of Japanese encephalitis virus in continuous cell lines. The study of the dynamics of fluorescence intensity permitted the objective evaluation of the previously revealed regularity in the increase of the intensity of the induced fluorescence of Japanese encephalitis antigen in continuous cell cultures.     

Latency to first naloxone-induced jump as a measure of precipitated abstinence intensity in morphine-dependent mice and the interaction of set on this test

A method developed in this laboratory uses latency to time of first jump after injection of naloxone rather than the number of jumps in a specified period or the number of animals jumping as a measure of the degree of morphine physical dependence. For the test, mice are placed in a glass cylinder used as a test chamber, after being injected with the antagonist. During the development of this method it was observed that repeated exposures of dependent mice to both naloxone and the chamber yielded shorter latencies to first jump than did repeated exposures to naloxone alone in animals with the same degree of physical dependence. It appears that learning develops when naloxone injections are given repeatedly and followed by exposure to the test chamber and that this learned behavior is manifested by a reduced latency to first jump which may be confused with increased intensity of the opiate-withdrawal syndrome.     

Kinetic resolution of a conformational transition and the ATP hydrolysis step using relaxation methods with a Dictyostelium myosin II mutant containing a single tryptophan residue

The fluorescence emission intensity from a conserved tryptophan residue (W501) located in the relay loop (F466 to L516) of the Dicytostelium discoideum myosin II motor domain is sensitive to ATP binding and hydrolysis. The initial binding process is accompanied by a small quench in fluorescence, and this is followed by a large enhancement that appears coincident with the hydrolysis step. Using temperature and pressure jump methods, we show that the enhancement process is kinetically distinct from but coupled to the hydrolysis step. The fluorescence enhancement corresponds to the open-closed transition (k(obs) approximately 1000 s(-1) at 20 degrees C). From the overall steady-state fluorescence signal and the presence or absence of a relaxation transient, we conclude that the ADP state is largely in the open state, while the ADP.AlF(4) state is largely closed. At 20 degrees C the open-closed equilibria for the AMP.PNP and ADP.BeF(x) complexes are close to unity and are readily perturbed by temperature and pressure. In the case of ATP, the equilibrium of this step slightly favors the open state, but coupling to the subsequent hydrolysis step gives rise to a predominantly closed state in the steady state. Pressure jump during steady-state ATP turnover reveals the distinct transients for the rapid open-closed transition and the slower hydrolysis step.     

The best warm-up for the vertical jump in college-age athletic men

The purpose of this study was to determine the effectiveness of specific and nonspecific warm-ups on the vertical jump test performed by athletic men. Twenty-nine men (18-23 years) in athletics (speed positions in football) performed vertical jump tests on 4 separate days after completing 4 different warm-up protocols. The 4 warm-up protocols were (a) submaximal jump warm-up, (b) weighted jump warm-up, (c) stretching warm-up, and (d) no warm-up. The weighted jump warm-up protocol required 5 countermovement jumps onto a box, with the athletes holding dumbbells equaling 10% of their body weight. The submaximal jump warm-up protocol required the athletes to perform 5 countermovement jumps at 75% intensity of their past maximum vertical jump score. The stretching warm-up protocol required the athletes to perform 14 different stretches, each held for 20 seconds. The no warm-up protocol required the athletes to perform no activity prior to being tested. Three vertical jumps were measured following each warm-up; the score for analysis was the best jump. The data were analyzed with a repeated measures analysis of variance and Bonferroni post hoc tests. The Bonferroni post hoc tests showed a significant difference (p < 0.001) between the weighted jump warm-up and all other warm-ups. The effect size was 0.380 and the power was 1.00 for the statistical analyses. We concluded that utilizing a weighted resistance warm-up would produce the greatest benefit when performing the vertical jump test.     

Enhanced fusion of a nucleopolyhedrovirus with cultured cells by a virus enhancing factor from an entomopoxvirus

Fusion of Pseudaletia unipuncta nucleopolyhedrovirus with an armyworm cell line (SIE-MSH-805-F) was studied by means of three fluorescence assays that are based on the relief of fluorescence self-quenching of octadecylrhodamine B chloride (R18). A gradual increase in fluorescence intensity indicative of virus-cell fusion was observed by spectrofluorometry when R18-labeled polyhedron-derived virus was incubated with cultured cells. The fusion was enhanced by the virus enhancing factor (EF) from Pseudaletia separata entomopoxvirus. Lysosomotropic agents had little effect on the virus-cell fusion. The percentage of positively fluorescent cells, as determined by flow cytometry, gradually increased after the addition of labeled virus and was higher in the presence of the EF than in its absence. Confocal microscopy of cultured cells that had been combined with labeled virus showed that the fluorescence appeared first on their surface. The plasma membrane of cultured cells had specific affinity to the EF, as revealed by indirect immunofluorescence microscopy.     

Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles

We present a single virion method to determine absolute distributions of copy number in the protein composition of viruses and apply it to herpes simplex virus type 1. Using two-color coincidence fluorescence spectroscopy, we determine the virion-to-virion variability in copy numbers of fluorescently labeled tegument and envelope proteins relative to a capsid protein by analyzing fluorescence intensity ratios for ensembles of individual dual-labeled virions and fitting the resulting histogram of ratios. Using EYFP-tagged capsid protein VP26 as a reference for fluorescence intensity, we are able to calculate the mean and also, for the first time to our knowledge, the variation in numbers of gD, VP16, and VP22 tegument. The measurement of the number of glycoprotein D molecules was in good agreement with independent measurements of average numbers of these glycoproteins in bulk virus preparations, validating the method. The accuracy, straightforward data processing, and high throughput of this technique make it widely applicable to the analysis of the molecular composition of large complexes in general, and it is particularly suited to providing insights into virus structure, assembly, and infectivity.     

The effect of the lipid bilayer state on fluorescence intensity of fluorescein-PE in a saturated lipid bilayer

Fluorescein-PE is a fluorescence probe that is used as a membrane label or a sensor of surface associated processes. Fluorescein-PE fluorescence intensity depends not only on bulk pH, but also on the local electrostatic potential, which affects the local membrane interface proton concentration. The pH sensitivity and hydrophilic character of the fluorescein moiety was used to detect conformational changes at the lipid bilayer surface. When located in the dipalmitoylphosphatidylcholine (DPPC) bilayer, probe fluorescence depends on conformational changes that occur during phase transitions. Relative fluorescence intensity changes more at pretransition than at the main phase transition temperature, indicating that interface conformation affects the condition in the vicinity of the membrane. Local electrostatic potential depends on surface charge density, the local dielectric constant, salt concentration and water organisation. Initial increase in fluorescence intensity at temperatures preceding that of pretransition can be explained by the decreased value of the dielectric constant in the lipid polar headgroups region related in turn to decreased water organisation within the membrane interface. The abrupt decrease in fluorescence intensity at temperatures between 25 degrees C and 35 degrees C (DPPC pretransition) is likely to be caused by an increased value of the electrostatic potential, induced by an elevated value of the dielectric constant within the phosphate group region. Further increase in the fluorescence intensity at temperatures above that of the gel-liquid phase transition correlates with the calculated decreased surface electrostatic potential. Above the main phase transition temperature, fluorescence intensity increase at a salt concentration of 140 mM is larger than with 14 mM. This results from a sharp decline of the electrostatic potential induced by the phosphocholine dipole as a function of distance from the membrane surface.     

Time-resolved imaging of HIV-1 Env-mediated lipid and content mixing between a single virion and cell membrane

A method has been developed to follow fusion of individual pseudotyped virus expressing HIV-1 Env to cells by time-resolved fluorescence microscopy. Viral envelopes were labeled with a fluorescent lipid dye (DiD) and virus content was rendered visible by incorporating a Gag-GFP chimera. The Gag-GFP is naturally cleaved to the much smaller NC-GFP fragment in the mature virions. NC-GFP was readily released upon permeabilization of the viral envelope, whereas the capsid was retained. The NC-GFP thus provides a relatively small and mobile aqueous marker to follow viral content transfer. In fusion experiments, virions were bound to cells at low temperature, and fusion was synchronously triggered by a temperature jump. DiD transferred from virions to cells without a significant lag after the temperature jump. Some virions released DiD but retained NC-GFP. Surprisingly, the fraction of lipid mixing events yielding NC-GFP transfer was dependent on the type of target cell: of three infectable cell lines, only one permitted NC-GFP transfer within minutes of raising temperature. NC-GFP release did not correlate with the level of CD4 or coreceptor expression in the target cells. The data indicate that fusion pores formed by HIV-1 Env can remain small for a relatively long time before they enlarge.     

Kinetics of conformational changes in tRNAPhe (yeast) as studied by the fluorescence of the y-base and of formycin substituted for the 3'-terminal adenine

The kinetics of the melting transitions of tRNA^phe (yeast) were followed by the fluorescence of the Y-base and of formycin substituted for the 3'-terminal adenine. As judged from differential UV absorbance melting cutves the formycin label had virtually no influence on the conformation of the tRNA. A temperature jump apparatus was modified to allow the simultaneous observation of transmission and fluorescence intensities by two independent optical channels. The design of a temperature jump cell with an all quartz center piece is given. The cell is resistant to temperatures up to 90°C; it provides high optical sensitivity, low stray light intensity and the possibility of measuring fluorescence polarization. The T-jump experiments allowed to discriminate between fast unspecific fluorescence quenching (τ <5 μsec) and slow co-operative conformational changes. In the central part of the temperature range of UV-melung (midpoint temperature 30°C in 0.01 M Na⁺ and 39°C in 0.03 M Na⁺, pH 6.8) two resolvable relaxation processes were observed. The coirssponding relaxation times were 20 msec and 800 msec at 30°C in 0.01 M Na⁺, and 4 msec and 120 msec at 39°C in 0.03 M Na⁺. The Y-base fluorescence shows both of the relaxation effects, which almost cancel in equilibrium fluorescence melting, because their amplitudes have opposite signs. From this finding the existence of some residual tertiary structure is inferred which persists after the unfolding of the main part of tertiary structure durirg early melting (midpoint temperature 24°C in 0.03 M Na⁺). In the fluorescence sigXXX of the formycin also the two relaxation effects appear. Both of them are connected with a decrease of the fluorescence intensity. From the results a coupled opening of the anticodon and acceptor branches is concluded.Enzymes: phenylalanyl-tRNA synthetase, PRS (EC 6.1.1.-20); ATP (CTP) tRNA nucleotidyl transferase, NT (EC 2.7.7.-20); alkaline phosphatase (EC 3-1-3.1).     

Conformational change of the foot protein of sarcoplasmic reticulum as an initial event of calcium release

Heavy sarcoplasmic reticulum vesicles were labeled with the thiol-reacting fluorescent probe N-(7-dimethylamino-4-methyl-4-coumarinyl)maleimide (DACM), and the DACM-labeled foot protein moiety was purified. The fluorescence intensity of the DACM attached to the foot protein decreased by the addition of low (activating) concentrations of ryanodine, while it increased at higher (inhibitory) concentrations, suggesting that the lower fluorescence represents the active state of the foot protein, while the higher fluorescence, its inactive state. Under conditions that induce Ca2+ release from SR (Ca2+ jump, addition of Ca2+ release inducing reagents such as caffeine and polylysine), the fluorescence intensity of the protein-attached DACM decreased rapidly (e.g. k congruent to 70 s-1 under optimum conditions). The initial rate of Ca2+ release from the DACM-labeled SR showed a close correlation with the amplitude of the fluorescence change of the foot protein-attached DACM under variety of conditions; e.g. in the presence of Ca2+, polylysine, ATP, and ruthenium red, etc. The fluorescence change of the foot protein was much faster than Ca2+ release from SR under a variety of conditions of Ca2+ release. We propose that the binding of release triggering reagents to the foot protein induces a rapid conformational change, which in turn regulates Ca2+ release.     

Evaluation of antiviral activity of different origin compounds by flow cytometry

Against many viral diseases caused for example by HSV, EBV, CMV, HIV, RSV, HCV for which vaccines are not available, chemiotherapeutics seem to have the principal significance. High progress in development of new antiviral compounds is observed. In addition to synthetic compounds a large number of naturally occurring substances have been shown to posses antiviral activity. One of such substance is tannic acid. In this study comparison of antiviral activity of tannic acid, acyclovir (ACV) and ganciclovir (GCV) against cytomegalovirus (CMV) is presented. The MRC5 cells infected with CMV and treated with different compounds were analyzed by flow cytometry and cythopatic effect inhibition test for inhibition of virus replication and by MTT assay for cytotoxity. It has been shown that tannic acid has antiviral activity against cytomegalovirus and that expression of virus antigens measured as median fluorescence intensity (MFI) by flow cytometry can be used for evaluation of virus replication.     

Magnetic protein microbead-aided indirect fluoroimmunoassay for the determination of canine virus specific antibodies

Rabies, canine distemper, and canine parvovirus are common contagious viral diseases of dogs and many other carnivores, and pose a severe threat to the population dynamics of wild carnivores, as well as endangering carnivore conservation. However, clinical diagnosis of these diseases, especially canine distemper and canine parvovirus, is difficult because of the broad spectrum of symptoms that may be confused with other respiratory and enteric diseases of dogs. The most frequently used and proven techniques for diagnosing viral diseases include the conventional enzyme-linked immunosorbent assay (ELISA), rapid fluorescent focus inhibition test (RFFIT), mouse neutralisation test (MNT), and fluorescent antibody virus neutralization (FAVN) test. However, these methods still have some inherent limitations. In this study, a magnetic protein microbead-aided indirect fluoroimmunoassay was developed to detect canine virus specific antibodies, human rabies immunoglobulin, CDV McAbs, and CPV McAbs. In this assay, an avidin-biotin system was employed to combine magnetic microbeads and virus antigens (rabies virus, canine distemper virus, and canine parvovirus). Quantification of the targeted virus antibodies was analyzed through indirect fluoroimmunoassay using the specific antigen-antibody reaction, as well as their corresponding FITC-labeled detection antibodies (mouse anti-human IgG/FITC conjugate or rabbit anti-dog IgG/FITC conjugate). The results indicated that the fluorescence intensity increased when a higher concentration of the targeted analyte was used, but the control had almost no fluorescence, much like the conventional ELISA. For human rabies immunoglobulin, CDV McAbs, and CPV McAbs, the minimum detectable concentrations were 0.2 IU/mL, 0.3 ng/mL, and 0.5 ng/mL, respectively. All of these results indicate that this assay can be employed to determine the presence of canine virus specific antibodies. In addition, the method devised here can be utilized as a general protocol in other bacterial and viral marker analysis.     

荧光定量RT-PCR检测甲型H1N1病毒核酸的临床意义

目的观察荧光定量RT-PCR技术在检测甲型H1N1病毒核酸的临床意义。方法对135例经确诊为甲型H1N1的感染患者的咽式子采用荧光定量PCR技术检测H1N1病毒核酸,同时对40例健康体检者的咽式子做为对照组一同进行甲型H1N1病毒核酸检测。结果 135例确诊为甲型H1N1的患者经荧光定量PCR检测阳性129例,符合率为96.99%。40例健康体检者结果全阴性。结论荧光定量PCR检测甲型H1N1病毒核酸具有快速、特异性高等特点,在采用此方法诊断甲型H1N1时,阳性即可确诊,阴性者要结合临床。     

Effect of aminoglycoside antibiotics on the conformation of the unpaired loop adenine of avian leucosis virus RNA as revealed using 2-aminopurine fluorescence

The fluorescent properties of 2-aminopurine (2-AP) incorporated in an RNA sequence are used to study the structural dynamics and local changes of the retroviral RNA structure. Using 2-AP, the conformational states of the unpaired loop adenine in avian leucosis virus RNA were studied upon its interaction with aminoglycoside antibiotics. The intensity of 2-AP fluorescence in the monomeric RNA hairpin was higher than in both RNA dimers. The intensity of fluorescence in the extended dimer was significantly lower than in the kissing loop dimer. The finding was be explained by the fact that stacking contacts in the extended dimer produce a more compact loop structure than in the kissing loop dimer. When the binding of aminogycoside antibiotics with the kissing loop dimer RNA was analyzed, only tobramycin increased the intensity of 2-AP fluorescence almost threefold. The results showed that 2-AP fluorescence is suitable for detecting local changes in complexes of retroviral RNA with ligands.     

The effect of short-term VertiMax vs. depth jump training on vertical jump performance

The ability to generate lower body explosive power is considered an important factor in many athletic activities. Thirty-one men and women, recreationally trained volunteers, were randomly assigned to 3 different groups (control, n = 10; VertiMax, n = 11; and depth jump, n = 10). A Vertec measuring device was used to test vertical jump height pre- and post-training. All subjects trained twice weekly for 6 weeks, performing approximately 140 jumps. The VertiMax group increased elastic resistance and decreased volume each week, while the depth jump group increased both box height and volume each week. The depth jump group significantly increased their vertical jump height (pre: 20.5 +/- 3.98; post: 22.65 +/- 4.09), while the VertiMax (pre: 22.18 +/- 4.31; post: 23.36 +/- 4.06) and control groups (pre: 15.65 +/- 4.51; post: 15.85 +/- 4.17) did not change. These findings suggest that, within the volume and intensity constraints of this study, depth jump training twice weekly for 6 weeks is more beneficial than VertiMax jump training for increasing vertical jump height. Strength professionals should focus on depth jump exercises in the short term over commercially available devices to improve vertical jump performance.     

High-throughput Detection Method for Influenza Virus

Influenza virus is a respiratory pathogen that causes a high degree of morbidity and mortality every year in multiple parts of the world. Therefore, precise diagnosis of the infecting strain and rapid high-throughput screening of vast numbers of clinical samples is paramount to control the spread of pandemic infections. Current clinical diagnoses of influenza infections are based on serologic testing, polymerase chain reaction, direct specimen immunofluorescence and cell culture ^1,2.Here, we report the development of a novel diagnostic technique used to detect live influenza viruses. We used the mouse-adapted human A/PR/8/34 (PR8, H1N1) virus ³ to test the efficacy of this technique using MDCK cells ⁴. MDCK cells (10⁴ or 5 x 10³ per well) were cultured in 96- or 384-well plates, infected with PR8 and viral proteins were detected using anti-M2 followed by an IR dye-conjugated secondary antibody. M2 ⁵ and hemagglutinin ¹ are two major marker proteins used in many different diagnostic assays. Employing IR-dye-conjugated secondary antibodies minimized the autofluorescence associated with other fluorescent dyes. The use of anti-M2 antibody allowed us to use the antigen-specific fluorescence intensity as a direct metric of viral quantity. To enumerate the fluorescence intensity, we used the LI-COR Odyssey-based IR scanner. This system uses two channel laser-based IR detections to identify fluorophores and differentiate them from background noise. The first channel excites at 680 nm and emits at 700 nm to help quantify the background. The second channel detects fluorophores that excite at 780 nm and emit at 800 nm. Scanning of PR8-infected MDCK cells in the IR scanner indicated a viral titer-dependent bright fluorescence. A positive correlation of fluorescence intensity to virus titer starting from 10²-10⁵ PFU could be consistently observed. Minimal but detectable positivity consistently seen with 10²-10³ PFU PR8 viral titers demonstrated the high sensitivity of the near-IR dyes. The signal-to-noise ratio was determined by comparing the mock-infected or isotype antibody-treated MDCK cells.Using the fluorescence intensities from 96- or 384-well plate formats, we constructed standard titration curves. In these calculations, the first variable is the viral titer while the second variable is the fluorescence intensity. Therefore, we used the exponential distribution to generate a curve-fit to determine the polynomial relationship between the viral titers and fluorescence intensities. Collectively, we conclude that IR dye-based protein detection system can help diagnose infecting viral strains and precisely enumerate the titer of the infecting pathogens.     

The acute effects of different stretching exercises on jump performance

The purpose of this study was to demonstrate the short-term effects of different stretching exercises during the warm-up period on the lower limbs. A controlled, crossover clinical study involving 49 volunteers (14 women and 35 men; mean age: 20.4 years) enrolled in a "physical and sporting activities monitor" program. The explosive force was assessed using the Bosco test. The protocol was as follows: The test involved a (pre) jump test, general warm-up, intervention and (post) jump test. Each volunteer was subjected to each of the 5 interventions (no stretching [NS] and stretching: static passive stretching [P]; proprioceptive neuromuscular facilitation [PNF] techniques; static active stretching in passive tension [PT]; static active stretching in active tension [AT]) in a random order. The jump test was used to assess the squat jump, countermovement jump (CMJ), elasticity index (EI), and drop jump. An intragroup statistical analysis was performed before and after each intervention to compare the differences between the different stretching exercises. An intergroup analysis was also performed. Significant differences (p < 0.05) were found between all variables for the interventions "P," "PNF," and "TA" in the intragroup analysis, with each value being higher in the postjump test. Only the "P" intervention showed a significant difference (p = 0.046) for "EI," with the postvalue being lower. Likewise, significant differences (p < 0.05) were observed for the "CMJ" measurements during the intergroup analysis, especially between "NS" and the interventions "P," "PNF," "AT," and "PT," with each value, particularly that for "AT," being higher after stretching. The results of this study suggest that static active stretching in AT can be recommended during the warm-up for explosive force disciplines.