Effects of growth regulators and incubation period on in vitro regeneration of adventitious shoots from gerbera petioles

An effective system for in vitro regeneration of adventitious shoots from callus for the transformation or mutation of gerbera was developed. Callus was produced from petioles of the youngest 3–4 leaves detached from auxillary shoots produced in vitro. Induction medium, on which leaves were incubated over 3 or 6 days, contained 2.3 μM thidiazuron and 0.53 μM α-naphthaleneacetic acid. Explants were than transferred to one of three regeneration media with lower levels of growth regulators. Regeneration was quantified over four (4-weeks each) passages at the time of explant transfer to fresh medium. Direct shoot regeneration occurred during the first 4 weeks, and after these shoots were discarded a semi-compact organogenic callus was produced. Effectiveness of shoot regeneration depended on four criteria: the cultivar (three cultivars were tested), the sequence of passage on regeneration medium, the growth regulators in regeneration medium and the duration of the induction period. Regeneration potential from calli of all cultivars increased from the first to the fourth passage. Duration of the incubation period on induction medium (3 or 6 days) influenced regeneration to varying degrees, depending on the cultivar used and the regeneration medium contents. There were no differences between two of the regeneration media – B, containing 2.2 μM 6-benzyladenine and 0.3 μM indole-3-acetic acid and C, containing 4.4 μM 6-benzyladenine, 4.6 μM zeatin and 0.6 μM indole-3-acetic acid. Cultivar Mariola was the most productive and regenerated more than seven shoots per callus in the fourth passage. Regeneration on medium B was further evaluated on four additional gerbera cultivars. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of magnetic fields on tumor growth and viability

Breast cancer is the most common nonskin cancer and is the second leading cause of cancer-related deaths in women. Most methods of intervention involve combinations of surgery, chemotherapy, and ionizing radiation. Both chemotherapy and ionizing radiation can be effective against many types of cancer, but they also harm normal tissues. The use of nonionizing, magnetic fields has shown early promise in a number of in vitro and animal studies. Our study tested the effect of varying durations of magnetic exposure on tumor growth and viability in mice injected with breast cancer cells. Cancer cells were labeled through stable expression of firefly luciferase for monitoring of tumor growth and progression by using an in vivo imaging system. We hypothesized that magnetic field exposure would influence tumor growth and progression. Our results showed that exposure of the mice to magnetic fields for 360 min daily for as long as 4 wk suppressed tumor growth. Our study is unique in that it uses an in vivo imaging system to monitor the growth and progression of tumors in real time in individual mice. Our findings support further exploration of the potential of magnetic fields in cancer therapeutics, either as adjunct or primary therapy.     

Effect of static magnetic fields on osteoblasts and fibroblasts in vitro

In vitro assays were made of the effect of a static magnetic field of a neodymium magnet on cellular behavior. The cell turnover rate was examined by the incorporation of radioactive thymidine, and anabolic processes were measured by the incorporation of radioactive proline. Cell cultures of fibroblast- and osteoblast-like cells of the neonatal rat calvarium were assayed to determine uptakes of radioactive thymidine and proline; these assays were performed in conjunction with examination of an explant of the rat calvarium. The cells were assayed after exposure to a field for 1-, 3-, 5-, 7-, and 10-day periods. Cells were exposed to north and south poles with a pole-face flux density of 0.61 T; control cultures were exposed to an unmagnetised piece of neodymium. After sham exposure or exposure to the magnetic field, 50 μCuries/ml of culture media of isotope were added to the culture medium. The cultures were returned to an incubator for 6 h. Then, following centrifugation, the supernatant was assayed for radioactivity in a scintillation counter after addition of 3 ml of scintillation fluid. A statistically significant magnetic stimulation of turnover rate and synthesis of fibroblasts was found, but stimulation of osteoblasts did not occur. Conversely, the explants, which represent the osteoblasts and fibroblasts in an organised system, showed a statistically significant inhibition in uptake of the radioactive label. The data indicate both variability and diversity of cellular behaviour, and they accentuate the need for caution in the interpretation of effects of static magnetic fields. © 1993 Wiley-Liss, Inc.     

Effects of 100 mT time varying magnetic fields on the growth of tumors in mice

The effects of 100-mT, 0.8-Hz square-wave magnetic fields on the growth of chemically induced tumors in mice were investigated. Tumors were initiated using one injection of benzo(a)pyrene (either 0.2 mg or 2.0 mg/animal). Male and female mice (Balb/c, C3H and C57/bl/6 strains) were exposed for 8 h/day from the onset of tumor until death or until the tumor volume reached a predetermined volume. Statistically significant decrease in the rate of tumor growth and increase in survival were observed in all cases. Results are discussed in terms of previous published work and of possible mechanisms.     

Exposure of maize seeds to stationary magnetic fields: Effects on germination and early growth

The effect of the exposure of maize seeds to stationary magnetic fields on germination and early growth has been studied under laboratory conditions. Seeds were magnetically exposed to one of two magnetic field strengths, 125 or 250 mT for different periods of time. Mean germination time and the time required to obtain 10, 25, 50, 75 and 90% of seeds to germinate were calculated. The results showed a reduction of these parameters for most of magnetic treatments, therefore their rate of germination was increased.Growth data measured on the 7th and 10th day after seeding allowed us to corroborate the effect observed in germination tests. Treated plants grew higher and heavier than control; on the 10th day total length was greater than control plants exposed to stationary magnetic field, corresponding with increase of the total fresh weight. The greatest increases were obtained for plants continuously exposed to 125 or 250 mT.     

Effect of permanent magnetic fields on the regeneration of the tail in amphibian larvae

Vertically directed permanent magnetic fields stimulate tail regeneration in larvae of three anuran species. This effect depends on correlation between the beginning of exposure and the time of injury, magnetic field intensity, species, age and individual features. The permanent magnetic field (133-193 kA/m) enhances differentiation of collagen-containing structures of the tail regenerate.     

Effects of static magnetic fields on diffusion in solutions

Static magnetic fields affect the diffusion of biological particles in solutions through the Lorentz force and Maxwell stress. These effects were analyzed theoretically to estimate the threshold field strength for these effects. Our results show that the Lorentz force suppresses the diffusion of charged particles such as Na+, K+, Ca2+, Cl-, and plasma proteins. However, the threshold is so high, i.e., more than 10(4) T, that the Lorentz force does not affect the ion diffusion at typical field strengths (a few Tesla at most). Since the threshold of gradient fields for producing a change in ion diffusion through the Maxwell stress is more than 10(5) T2/m for paramagnetic molecules (FeCl3, O2) and plasma proteins, their diffusion would be unaffected by typical gradient fields (100 T2/m at most) and even by high gradient fields (less than 10(5) T2/m) used in magnetic separation techniques. In contrast, movement of deoxygenated erythrocytes and FeCl3 colloids (more than 10(3) molecules) is influenced by the usual gradient fields due to a volume effect.     

Effects of magnetic fields on fyke net performance

The effects of magnetic fields generated by ferritic magnets and mounted in entrances to fyke nets were studied in 1999–2002. With respect to all fish species trapped (perch, pike, roach, rudd, bleak, bream, ruffe) except eel, the catch potential of magnet‐rigged fyke nets was found to be significantly higher (almost 50% on average) when compared with controls. Differences in magnet placement (N or S poles inside the traps) proved to have no significant importance on the number of fish that chose to enter the fyke nets. Specific behavioural mechanisms in adult fish towards magnetic fields as well as application of fishing gear magnet rigging in fisheries practice in inland waters are discussed.     

Effects of extremely-low-frequency magnetic fields on biological magnetite

Adair [Bioelectromagnetics 14:1–4, 1993] writes that “the effects of 60 Hz magnetic fields of 5 μT (50 mG) or less on biological structures holding magnetite (Fe₃O₄) are shown to be much smaller than those from thermal agitation; hence such interactions cannot be expected to be biologically significant.” This conclusion is questioned, because it appears to be based on a model that probably has very limited validity for pertinent biological systems. Furthermore, biologically plausible parameters can be selected to show that even this particular model does not exclude biologically significant effects of 60 Hz magnetic fields below 5 μT. Reported experimental results indicate effects in mammals of 50 Hz fields at the 1 μT level. © 1994 Wiley-Liss, Inc.     

Effects of ELF magnetic fields on biological magnetite

The effects of 60 Hz magnetic fields of 5 μT (50 mG) or less on biological structures holding magnetite (Fe₃O₄) are shown to be much smaller than that from thermal agitation; hence such interactions cannot be expected to be biologically significant. © 1993 Wiley-Liss, Inc.     

Effect of boron and methionine on growth and ion content in kiwifruit shoots cultured in vitro

The growth of kiwifruit explants was affected by boron (B) and methionine (Meth) in the culture medium. The longest shoots, the greatest number of shoots and the highest amount of fresh mass per explant were produced in Murashige and Skoog medium with 2 mM B and 2 μM Meth. Furthermore, by increasing B concentration in the culture medium from 0 to 2 mM, an increased rate of shoot proliferation was observed for the various Meth concentrations employed.     

Factors influencing the growth of micropropagated shoots and in vitro flowering of gentian

About 70% of the shoots developed from nodal explants ofGentiana triflora flowered in vitroondouble strength WPM medium containing 3% (w/v) sucrose, 0.5mg/l BA after 12 weeks of culture in a growth room at 22°Cwith continuous illumination (PPFD=60molm^–2 s^–1). The influences oninvitro shoot development and flowering of several factors includingthe position of the explant, requirements for sucrose, cytokinin orGA₃, variations of pH and photosynthetic photon flux density (PPFD)were investigated. In vitro flowering but not shootdevelopment of G. triflora decreased notably withincreaseddistance from the apex of the shoot, indicating the presence of a floralgradient in the micropropagated shoots. Conversely, as little as 0.01mg l^–1 GA₃ in the medium promotedshootdevelopment but even up to 0.2 mg l^–1GA₃ did not induce in vitro flowering.Even though BA could substitute GA₃ for a high level of shootdevelopment, it also promoted a high level of in vitroflowering at the PPFD of 60 molm^–2 s^–1. Sucrose was required for shootdevelopment and flowering in vitro and higher levels ofPPFD could not compensate effectively for the omission of the sugar from themedium. In general, the effects of different concentrations of BA in the mediumor variations of pH on shoot development and flowering invitro were found to be influenced by PPFD. A novel observation isthat precocious flowering of micropropagated gentian shoots did not occur ifthey were first cultured for 5 weeks in the dark before transfer to the lightcondition.     

Effects of low-frequency magnetic fields on fetal development in rats

We studied effects of alternating magnetic fields on the embryonic and fetal development of rats. Mated females of the Han:Wistar-strain were sham exposed or exposed continuously to a 50-Hz field or to a 20,000 pulse-per-second (pps) sawtooth magnetic field from day 0 to day 20 of pregnancy for 24 h/day until necropsied on day 20. The respective peak-to-peak intensities of the fields were 35.6 μT (sinewave) and 15.0 μT (sawtooth). Each treatment group contained 72 bred females. Control animals were kept under the same conditions without the magnetic field. No adverse effects were seen in the dams. The mean numbers of implantations and living fetuses per litter were statistically significantly increased in the 50-Hz group. There were, however, three total resorptions of litters in dams of the control group, which contributed to the difference in the number of living fetuses. The corrected body-mass gains (gains without uterine content) of dams were similar in all groups. Pregnancy rates, incidences of resorptions. late fetal deaths, and fetal body masses were similar in all groups. The incidence of fetuses with minor skeletal anomalies was statistically significantly increased in both exposed groups. Only one serious malformation (anophthalmia, sawtooth-exposed group) and a few minor visceral malformations were found. In conclusion, the magnetic fields used in this study did not increase the incidence of major malformations or resorptions in Wistar rats. The increased number of skeletal anomalies and implantations we observed indicates, however, that some developmental effects in rats may attend exposure to time-varying magnetic fields. © 1993 Wiley-Liss. Inc.     

Hydrolysis of various peptides and proteins in weak permanent and low frequency fluctuating magnetic fields

It was shown that weak combined magnetic fields (constant field 25-130 microT; variable field 0.01-0.2 microT; the range of effective frequencies of the alternating component 1-10 Hz) substantially increase the rate of hydrolysis of some proteins and peptides (eight various sequences). The concentration dependence of the dynamics of the process and the dependence of the magnitude of the effect on the parameters of the magnetic field. It was found that: (1) the effect is transmitted through a solvent preliminarily treated by magnetic fields and (2) the effect occurs in the presence of inhibitors of proteases and enzymes inactivating peroxides (catalase and horse radish peroxidase with substrate).     

Homogeneous magnetic fields influence pancreatic islet function in vitro

Pancreatic islets were isolated from newborn rats and exposed to homogeneous magnetic fields for 48 hours. Under these conditions insulin release, measured at low (5.6 mM) and high (16.7 mM) glucose concentrations, was significant and dose-dependent only at low glucose concentrations. High glucose and aminophylline (10 mM) inhibited insulin release. Thus, in the absence of stimulatory glucose concentrations, low-intensity magnetic fields (1 to 10 Gauss) significantly influence insulin discharge from rat islets in vitro.     

Effects of high-frequency electromagnetic fields on human lymphocytes in vitro

Human peripheral lymphocytes were incubated in the presence of high-frequency electromagnetic fields of 380, 900 and 1800 MHz. The measured endpoints were cell cycle progression and the frequencies of sister-chromatid exchanges. No differences between treated and control cultures could be found.     

地生兰(Cymbidium)个体发生途径研究

地生兰的类原球茎体(PLB)在不加激素的1/2 MS培养基上驯化培养,获得“无需外源激素仍能增殖的无性系(PLB-O)”,已继代5年,不分化芽和根。连续光照有利于PLB-O分生区的增殖,8h光照的次之,黑暗中增殖速度最低。PLB培养在含10％椰乳的1/2 BMS液体培养基中,随继代培养进程,形成分枝更多的丛生型PLB,约4个月后,在PLB顶端开始形成植株。兰花离体培养中,在同一个培养物中,同时存在着处于不同发育时期的分生区、原球茎、芽和小植株。PLB培养在含1/2BMS+6BA 2mg/L+NAA 0.2 mg/L的培养基中,同一个丛生型PLB上,因分生区和PLB的发育年龄的不同,形成不同发育年龄的幼芽。     

Effects of pulsed magnetic fields on the developing mouse embryo

The influence of a pulsed magnetic field (PMF; sawtooth with 45-μs linear rise time and 5-μs decay time, peak strength of 15 μT, and frequency 20 pps) on the embryogenesis of CBA/S mice was investigated in five experiments based on a total of 707 exposed and 543 unexposed primigravidas. Sham and PMF exposures began on day 1 of gestation (experiments 1 and 2), on day 2 (experiment 3), on day 5 (experiment 4). and on day 7 (experiment 5): all exposures continued until day 19 post conception (p.c.) when they were terminated, at which time the following variables were measured: number of implants; number of placental resorptions; number of living fetuses; number of dead fetuses; number of malformations in living and dead fetuses; and length and body mass of living fetuses. Control dams were sham-exposed concurrently with corresponding. PMF-exposed dams. With the exception of experiment 5, in which exposure to PMF started on day 7 p.c., all groups of exposed mice had significantly more placental resorptions when compared with concurrent controls. The increased resorption rate was not reflected in a reduction in litter size or in the number of litters. A significant increase in malformed fetuses was not seen in any of the exposed groups, or when groups were pooled. Only in experiment 1 was the number of dead fetuses affected by exposure to PMF. The effect of PMF on the implantation rate was not significant. Body mass and length of exposed fetuses were significantly reduced only when the PMF treatment began on day 7 p.c. That PMF-treated mice had significantly more placental resorptions when exposure began on day 5 p.c. or earlier (before implantation), but not when exposure began on day 7 (after implantation), may indicate a causative pre-implantation effect. Because a PMF-induced increase in the number of resorptions has not been observed in other strains of mice, the effect might be strain-related. © 1993 Wiley-Liss, Inc.     

Effects of very weak magnetic fields on radical pair reformation

We can expect that biological responses to very weak ELF electromagnetic fields will be masked by thermal noise. However, the spin of electrons bound to biologically important molecules is not strongly coupled to the thermal bath, and the effects of the precession of those spins by external magnetic fields is not bounded by thermal noise. Hence, the known role of spin orientation in the recombination of radical pairs (RP) may constitute a mechanism for the biological effects of very weak fields. That recombination will generally take place only if the valence electrons in the two radicals are in a singlet state and the effect of the magnetic field is manifest through differential spin precessions that affect the occupation of that state. Because the spin relaxation times are of the order of microseconds, any effects must be largely independent of frequency up to values of a few megahertz. Using exact calculations on an appropriately general model system, we show that one can expect small, but significant, modifications of the recombination rate by a 50 microT field only under a narrow range of circumstances: the cage time during which the two elements are together must be exceptionally long--of the order of 50 ns or longer; the hyperfine field of either radical must not be so great as to generate a precession period greater than the cage containment time; and the characteristic recombination time of the radical pair in the singlet state must be about equal to the containment time. Thus, even under such singularly favorable conditions, fields as small as 5 microT (50 milligauss) cannot change the recombination rate by as much as 1%. Hence, we conclude that environmental magnetic fields much weaker than the earth's field cannot be expected to affect biology significantly by modifying radical pair recombination probabilities.