Electronic paramagnetic resonance investigation of the activity of Origanum vulgare L. essential oil on the Listeria monocytogenes membrane.

AIMS: To evaluate the effect of oregano essential oil on Listeria monocytogenes cytoplasmic membrane. METHODS AND RESULTS: Nitroxide free-radical Electron Paramagnetic Resonance was applied on L. monocytogenes after 30 min exposure to oregano essential oil concentrations ranging from 0 to 1.25%. The impact of essential oil on the number of viable cells was evaluated by plate count. Growth dynamics of survivors in BHI and TSB were evaluated by turbidometry. After exposure to essential oil concentrations up to 0.50%, the membrane fluidity was changed and its order increased. When L. monocytogenes was exposed to higher concentrations, membrane order parameters slightly returned to the values of untreated cells. However, when the cells were exposed to EO in the presence of sodium azide, which impairs energy metabolism, the membrane fluidity was progressively enhanced, even at the lowest EO concentration (0.25%). Microbiological analyses confirmed a progressive reduction of viable count, at increasing essential oil concentrations. Both in BHI and TSB, the Lag phase length increased in treated cells with respect to controls, suggesting a cell damage recovery. CONCLUSIONS: The combined approach including microbiological and EPR analyses provided relevant information on membrane modification and cell response to essential oils. SIGNIFICANCE AND IMPACT OF THE STUDY: EPR approach was demonstrated to be an effective and helpful tool to comprehend the modifications exerted by essential oil on the bacterial membrane.

Extremely low frequency electromagnetic field exposure affects fertilization outcome in swine animal model.

Modern society continuously exposes the population to electromagnetic radiation, the effects of which on human health, in particular reproduction, are still unknown. The aim of this research was to assess the effect of acute (1h) exposure of boar spermatozoa to a 50 Hz extremely low frequency electromagnetic field (ELF-EMF) on early fertility outcome. The effect of intensities ranging from 0 to 2 mT on morpho-functional integrity of capacitated spermatozoa was examined in vitro. The oviducts containing or without spermatozoa were then exposed to the minimum in vivo, TD(50,) and maximum intensities determined in vitro, 4h before ovulation. The effects of ELF-EMF on spermatozoa in terms of early embryo development were evaluated after 12h and 6 days. It was found that in vitro ELF-EMF > 0.5 mT induced a progressive acrosome damage, thus compromising the ability of spermatozoa to undergo acrosomal reaction after zona pellucida stimulation and reducing the in vitro fertilization outcome. These effects became evident at 0.75 mT and reached the plateau at 1 mT. Under in vivo conditions, the ELF-EMF intensity of 1 mT was able to compromise sperm function, significantly reducing the fertilization rate. In addition, the exposure of oviducts to fields > or = 0.75 mT in the absence of spermatozoa was able to negatively affect early embryo development. In fact, it was found to cause a slowdown in the embryo cleavage. In conclusion, it was demonstrated how and at which intensities ELF-EMF negatively affect early fertility outcome in a highly predictive animal model.

Effects of 50 Hz extremely low frequency magnetic field on the morphology and function of boar spermatozoa capacitated in vitro.

The aim of this study was to evaluate the effect of an acute exposure to a sinusoidal MF-ELF (50 Hz, 1mT) on the ability of boar mature spermatozoa to acquire the fertilizing competence in vitro. The spermatozoa exposed during the 4h of incubation to the MF-ELF were evaluated for morphological (surface morphology and acrosome integrity) and functional parameters (cell viability, motility, induction of acrosomal reaction, AR, and the ability to in vitro fertilize oocytes). In parallel, the intracellular Ca(2+) levels as well as the major mechanisms of Ca(2+) clearance were assessed: (45)Ca intakes and intracellular Ca(2+) sequestration by analyzing intracellular Ca(2+) elevation induced by thapsigargin or studying mitochondrial function with Mito-Tracker. The MF-ELF exposure did not affect sperm viability and morphology during the first h of incubation when sperm Ca(2+) homeostasis were already compromised. First of all, MF-ELF treated spermatozoa showed resting intracellular Ca(2+) levels significantly lower than those recorded in controls. This result was dependent on a lower extracellular Ca(2+) intake and from the inhibitory role exerted on both intracellular Ca(2+) storages. As a consequence, after 1h of incubation MF-ELF exposed cells displayed a reduced motility, a modest reactivity when coincubated with solubilized zonae pellucidae and a reduction in oocyte penetrating ability. After 2 or 4h of incubation, in addition, signs of morphological damage appeared on plasma membrane and at acrosomal level. In conclusion, MF-ELF influence negatively spermatozoa first by impairing cell Ca(2+) homeostasis then by dramatically affecting sperm morphology and function.

A preliminary study of oscillating electromagnetic field effects on human spermatozoon motility.

Some effects of extremely low frequency electromagnetic fields (ELF-EMFs) on human spermatozoa are reported. Significant increases in the values of the motility and of the other kinematic parameters have been observed when spermatozoa were exposed to an ELF-EMF with a square waveform of 5 mT amplitude and frequency of 50 Hz. By contrast, a 5 mT sine wave (50 Hz) and a 2.5 mT square wave (50 Hz) exposure did not produce any significant effect on sperm motility. The effects induced by ELF-EMF (50 Hz; 5 mT) during the first 3 h of exposure persisted for 21 h after the end of the treatment. These results indicate that ELF-EMF exposure can improve spermatozoa motility and that this effect depends on the field characteristics.

Conformational changes at the active site of pantetheine hydrolase during denaturation by guanidine hydrochloride.

Conformational changes at the active site of pantetheine hydrolase (EC3.5.1.-) during guanidine hydrochloride (GndHCl) denaturation were investigated by UV and circular dichroism spectroscopy and by electron spin resonance spectroscopy, following the spectral behaviour of the nitroxide radicals (N-(1-oxyl-2,2,5,5,-tetramethyl-3-pyrrolidinyl) iodacetamide) covalently linked to the two active site cysteine residues. At low denaturant concentrations (0.2 M) no conformational changes may be observed, whereas the catalytic activity, is strongly affected. The results indicate that the active site of pantetheine hydrolase is labile and unfolds under conditions in which no global tertiary structure modifications can be observed.

Comparison of Gd-BOPTA with Gd-DTPA in MR imaging of rat liver.

A new lipophilic compound, gadolinium benzyloxypropionictetraacetate (BOPTA), with a high rate of biliary excretion was assessed as a magnetic resonance (MR) hepatospecific contrast-enhancing agent and compared with Gd-DTPA (diethylenetriaminepentaacetic acid) in MR imaging of normal rats. T1-weighted spin-echo images obtained before and after administration of each contrast agent at doses of 0.25, 0.5, and 1.0 mmol/kg showed greater enhancement of the liver with Gd-BOPTA than with Gd-DTPA, with the advantage more evident at lower doses. Images obtained with an inversion recovery sequence at the null value of rat liver parenchyma after injection of 0.1- and 0.5-mmol/kg doses of the contrast agent provided better evidence of the greater and longer-lasting hepatic enhancement due to Gd-BOPTA when compared with that of Gd-DTPA. Gd-BOPTA is a potentially good contrast agent for obtaining prolonged enhancement of the liver, permitting studies during the long time needed to acquire conventional T1-weighted images.

[An animal model for the study of liver regeneration by magnetic resonance imaging]. / Modello animale per lo studio della rigenerazione epatica in Risonanza Magnetica per Immagini.

Liver regeneration after partial hepatectomy was studied in rats by means of magnetic resonance (MR) imaging and T1 relaxation time. Fourteen hepatectomized rats were compared to sham operated ones and to controls which had not undergone any surgical treatment. The animals were imaged at 0.5 T, and inversion recovery (IR) technique was employed at the T-null of liver before surgery. T1 was determined in vitro with spectroscopy. Regenerating rat liver exhibited a significantly high increase in MR signal intensity and T1 values (p less than 0.05) 24 hours after surgery, returning to baseline values at 2 weeks. Sham operated animals and controls did not exhibit significant changes in signal intensity from baseline values (p greater than 0.05). These findings suggest that MR imaging is able to detect the pathophysiological changes occurring in liver parenchyma during the regenerating process and to monitor different stages of the hyperplastic process.

[A comparative study between Gd-BOPTA, a biliary excretion contrast medium, and Gd-DTPA in the magnetic resonance imaging of the rat liver]. / Studio comparativo tra Gd-BOPTA, mezzo di contrasto ad escrezione biliare, e Gd-DTPA nell'imaging a Risonanza Magnetica del fegato di ratto.

A new lipophilic compound, Gd-BOPTA, presenting a high rate (38.6%) of biliary excretion was tested as an hepato-specific MR contrast agent. Its adequacy was compared to that of Gd-DTPA in laboratory animals. T1-weighted spin-echo sequences (TR 220 ms, TE 20 ms) both before and after the administration of the 2 contrast agents (doses: 0.25, 0.5, and 1.0 mmol/kg) showed better liver enhancement with Gd-BOPTA than with Gd-DTPA. Gd-BOPTA superiority was more evident at lower doses, while at 1.0 mmol/kg a comparable enhancement was achieved. Inversion recovery sequence at the T-null of liver parenchyma before contrast (TR 800 ms, TE 30 ms, TI 100 ms) was performed after the injection of 0.1 and 0.5 mmol/kg of Gd-DTPA and Gd-BOPTA. This sequence allowed the good and long-lasting liver enhancement achieved with Gd-BOPTA to be even better demonstrated, while Gd-DTPA provided only a slight and early enhancement with 0.1 mmol/kg and returned to baseline values 60' after the injection of the highest dose (0.5 mmol/kg). Gd-BOPTA proved to be a good contrast agent to obtain prolonged liver enhancement, thus providing the radiologist with the long time needed to acquire conventional T1-weighted pulse sequences.

MR characterization of hepatic lesions by t-null inversion recovery sequence.

Tissue characterization of focal hepatic lesions was performed employing an inversion recovery sequence with short repetition time and short inversion time (TI) values. Different and specific TI values, correlated to the in vivo measured T1 relaxation times, were used to null the signal intensity of each type of lesion. In 40 patients studied, we observed the nulling effect of normal liver in 10 of 10 cases with a TI of 136 ms, of metastases in 7 of 8 cases with a TI of 175 ms, of hemangiomas in 15 of 16 cases with a TI of 200 ms, and of cysts in 6 of 7 cases with a TI of 235 ms. A quantitative analysis of the nulled signal was performed by measuring the signal/noise values. A further qualitative and quantitative characterization was carried out by evaluating the signal intensity of hepatic lesions at the null point of normal liver. The method provided the possibility of discriminating different focal lesions with specificity values of 83-94%, according to the type of lesion.