Extremely low frequency electromagnetic fields modulate expression of inducible nitric oxide synthase, endothelial nitric oxide synthase and cyclooxygenase-2 in the human keratinocyte cell line HaCat: potential therapeutic effects in wound healing.

BACKGROUND: Extremely low frequency (ELF) electromagnetic fields (EMF) are known to produce a variety of biological effects. Clinical studies are ongoing using EMF in healing of bone fractures and skin wounds. However, little is known about the mechanisms of action of ELF-EMF. Several studies have demonstrated that expression and regulation of nitric oxide synthase (NOS) and cyclooxygenase-2 (COX-2) are vital for wound healing; however, no reports have demonstrated a direct action of ELF-EMF in the modulation of these inflammatory molecules in human keratinocytes. OBJECTIVES: The present study analysed the effect of ELF-EMF on the human keratinocyte cell line HaCaT in order to assess the mechanisms of action of ELF-EMF and to provide further support for their therapeutic use in wound healing. METHODS: Exposed HaCaT cells were compared with unexposed control cells. At different exposure times, expression of inducible NOS (iNOS), endothelial NOS (eNOS) and COX-2 was evaluated by Western blot analysis. Modulation of iNOS and eNOS was monitored by evaluation of NOS activities, production of nitric oxide (NO) and O(2)(-) and expression of activator protein 1 (AP-1). In addition, catalase activity and prostaglandin (PG) E(2) production were determined. Effects of ELF-EMF on cell growth and viability were monitored. RESULTS: The exposure of HaCaT cells to ELF-EMF increased iNOS and eNOS expression levels. These ELF-EMF-dependent increased expression levels were paralled by increased NOS activities, and increased NO production. In addition, higher levels of AP-1 expression as well as a higher cell proliferation rate were associated with ELF-EMF exposure. In contrast, ELF-EMF decreased COX-2 expression, PGE(2) production, catalase activity and O(2)(-) production. CONCLUSIONS: Mediators of inflammation, such as reactive nitrogen and PGE(2), and keratinocyte proliferation are critical for the tissue regenerative processes. The ability of ELF-EMF to upmodulate NOS activities, thus nitrogen intermediates, as well as cell proliferation, and to downregulate COX-2 expression and the downstream intermediate PGE(2), highlights the potential therapeutic role of ELF-EMF in wound healing processes.

Extremely low frequency electromagnetic field enhances human keratinocyte cell growth and decreases proinflammatory chemokine production.

BACKGROUND: Proliferation and differentiation of keratinocytes are central processes in tissue regeneration after injury. Chemokines, produced by a wide range of cell types including keratinocytes, play a regulatory role in inflammatory skin diseases. Several studies have shown that an electromagnetic field (EMF) can influence both inflammatory processes and repair mechanisms including wound healing on different tissue models. OBJECTIVES: To elucidate the effect of extremely low frequency EMF (ELF-EMF) on keratinocyte proliferation and production of chemokines [RANTES, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1 alpha and interleukin (IL)-8] in order to evaluate a potential therapeutic use of magnetic fields. METHODS: The human keratinocyte cell line HaCaT was exposed at 1 mT, 50 Hz for different lengths of time and compared with unexposed control cells. Cell growth and viability were evaluated at different exposure times by cell count and trypan blue exclusion. Chemokine production and expression were analysed by enzyme-linked immunosorbent assay (ELISA) and by real-time polymerase chain reaction. Total NF-kappaB p65 was quantified by ELISA. RESULTS: Significantly increased growth rates were observed after 48 h of EMF exposure as compared with control cells, while no difference in cell viabilities were detected. Gene expression and release of RANTES, MCP-1, MIP-1 alpha and IL-8 were significantly reduced after 72 h of exposure. NF-kappaB levels became almost undetectable after only 1 h of EMF exposure, and were inversely correlated with cell density. CONCLUSIONS: Our results show that ELF-EMF modulates chemokine production and keratinocyte growth through inhibition of the NF-kappaB signalling pathway and thus may inhibit inflammatory processes. ELF-EMF could represent an additional therapeutic approach in the treatment of skin injury.

Combined effects of electromagnetic fields on immune and nervous responses.

In technologically developed countries, there is concern about hazards from electromagnetic fields (EMFs). Several studies have reported that immune and neuroendocrine systems exert an integrated response to EMF exposure. The aim of this review is to summarize the results of studies on the effect of low and high frequency EMF on immune and neuroendocrine systems on which our research group has been working for several years.

Regional changes in the metabolite profile after long-term hypoxia-ischemia in brains of young and aged rats: a quantitative proton MRS study.

Quantitative proton magnetic resonance spectroscopy (MRS) was used to determine region-specific metabolic changes in young and aged animals subjected to a long-term hypoxic-ischemic injury. Focal ischemia, which was studied as an experimental stroke model, was induced in 3- and 24-month-old rats by unilateral common carotid artery occlusion associated with 24 h of hypoxia. Eight metabolites were quantified from extracts in three different brain regions (hippocampus, frontoparietal and occipital cortices) from both the ipsilateral and contralateral sides. Our findings showed significant differences in lactate and myo-inositol concentration values in the hippocampus of the aged rats as compared to the same area of the young adult group under normoxic conditions. After hypoxia-ischemia (HI), the most relevant changes in metabolite concentrations were found in the hippocampal region of both young and aged groups as compared to their age-matched controls. Of the three brain areas under investigation, the hippocampus proved to be particularly susceptible to the prolonged hypoxia-ischemia perturbation. The effects were more evident in the aged animals.

Optimal filter design for shielded and unshielded ambient noise reduction in fetal magnetocardiography.

The greatest impediment to extracting high-quality fetal signals from fetal magnetocardiography (fMCG) is environmental magnetic noise, which may have peak-to-peak intensity comparable to fetal QRS amplitude. Being an unstructured Gaussian signal with large disturbances at specific frequencies, ambient field noise can be reduced with hardware-based approaches and/or with software algorithms that digitally filter magnetocardiographic recordings. At present, no systematic evaluation of filters' performances on shielded and unshielded fMCG is available. We designed high-pass and low-pass Chebychev II-type filters with zero-phase and stable impulse response; the most commonly used band-pass filters were implemented combining high-pass and low-pass filters. The achieved ambient noise reduction in shielded and unshielded recordings was quantified, and the corresponding signal-to-noise ratio (SNR) and signal-to-distortion ratio (SDR) of the retrieved fetal signals was evaluated. The study regarded 66 fMCG datasets at different gestational ages (22-37 weeks). Since the spectral structures of shielded and unshielded magnetic noise were very similar, we concluded that the same filter setting might be applied to both conditions. Band-pass filters (1.0-100 Hz) and (2.0-100 Hz) provided the best combinations of fetal signal detection rates, SNR and SDR; however, the former should be preferred in the case of arrhythmic fetuses, which might present spectral components below 2 Hz.

Integrated software suite for magnetocardiographic data analysis--a proposal based on an interactive programming environment.

OBJECTIVES: This paper describes an integrated software suite (ISS) for the processing of magnetocardiographic (MCG) recordings obtained with super-conducting multi-channel systems having different characteristics. We aimed to develop a highly flexible suite including toolboxes for current MCG applications, organized consistently with an open architecture that allows function integrations and upgrades with minimal modifications; the suite was designed for the compliance not only of physicists and engineers but also of physicians, who have a different professional profile and are accustomed to retrieve information in different ways. METHODS: The MCG-ISS was designed to work with all common graphical user interface operative systems. MATLAB was chosen as the interactive programming environment (IPE), and the software was developed to achieve usability, interactivity, reliability, modularity, expansibility, interoperability, adaptability and graphics style tailoring. Three users, already experienced in MCG data analysis, have intensively tested MCG-ISS for six months. A great amount of MCG data on normal subjects and patients was used to assess software performances in terms of user compliance and confidence and total analysis time. RESULTS: The proposed suite is an all-in-one analysis tool that succeeded in speeding MCG data analysis up to about 55% with respect to standard reference routines; it consequently enhanced analysis performance and user compliance. CONCLUSIONS: Those results, together with the MCG-ISS advantage of being independent on the acquisition system, suggest that software suites like the proposed one could uphold a wider diffusion of MCG as a diagnostic tool in the clinical setting.

Automatic detection of cardiac waves on fetal magnetocardiographic signals.

Fetal magnetocardiography (fMCG) provides fetal cardiac traces useful for the prenatal monitoring of fetal heart function. In this paper, we describe an analytical model (ACWD) for the automatic detection of cardiac waves boundaries that works on fetal signals reconstructed from fMCG by means of independent component analysis. ACWD was validated for 45 healthy and 4 arrhythmic fetuses ranging from 22 to 37 weeks; ACWD outcomes were compared with the estimates of three independent investigators. Descriptive statistics were used to assess correspondence between the outcomes of the automatic and manual approaches. The parametric two-tailed Pearson correlation test (alpha=0.01) was employed to quantify, by means of the coefficients of determination, the amount of common variation between the sequences of intervals quantified automatically and manually. ACWD performances on short and long rhythm strips were investigated. ACWD demonstrated to be a robust tool providing dependable estimates of cardiac intervals and their variability during the third gestational trimester also in case of fetal arrhythmias. SNR and stability of fetal traces were the factors limiting ACWD performances. ACWD computation time, which was approximately 1:600 with respect to the manual procedure, was comparable with the time required for fCTI estimation on averaged beats.

Effect of electromagnetic fields on several CD markers and transcription and expression of CD4.

We carried out flow cytometric analysis for multiparametric evaluation of cell surface markers related to cellular functions. Specifically, we studied the expression of CD4, CD8, CD3, CD16, CD19, HLA-DR, and CD14 macrophage receptors expression and cell cycle progression on cells exposed to ELF-EMF. In addition, we tested the effects of ELF-EMF on CD4 mRNA protein transcription and translation and the cell-cycle progression using an immunofluorescence method. Our data show that same CD surface marker expression are weakly influenced by electromagnetic fields, with no differences between cells exposed or not exposed to ELF-EMFs. However, when the CD4 protein generation was studied, an indication of protein production was found in lymphocytes exposed to ELF-EMF, as evidenced by immunofluorescence, Western blotting and RT-PCR analysis. CD16 and CD14 expression were affected by EMF exposure at all times studied (24, 48, 72 h). The results obtained with cell cycle analysis show that after 48 h of exposure to ELF-EMF, PHA-activated and not activated cells in S phase increase with respect to non-exposed cells. The findings from this study demonstrate that under our defined experimental conditions there is evidence that ELF-EMF has a slight effect on CD4, CD14 and CD16 receptor expression, while the other CD receptors are not affected.

The validation of volumetric real-time 3-dimensional echocardiography for the determination of left ventricular function.

The objective of this study was to validate a real-time 3-dimensional echocardiography (3DE) technique for the determination of left ventricular (LV) volume and ejection fraction (EF). In 10 mongrel dogs, an electromagnetic flow (EMF) probe was placed on the aorta, and the thorax was closed. Transthoracic imaging was performed during multiple hemodynamic conditions (n = 58) with simultaneous measurement of stroke volume (SV) with the use of EMF. From the volumetric data set, LV volumes were manually traced off-line by 2 independent observers with an apical rotation method (6 planes) and a conventional method (biplane) in a subset of conditions. This tracing technique was also evaluated in 18 human subjects in whom the calculated EF values were compared with values derived by multigated radionuclide angiography (MUGA). Excellent correlation and close limits of agreement were noted between SV measured by 3DE and EMF (r = 0.93) in dogs. In comparison with EMF-derived SV, 3DE provided better correlation than the biplane method (r = 0.93 versus r = 0.61). Interobserver and intraobserver variabilities were comparable (r = 0.94 and r = 0.94, respectively). In a comparison of MUGA-derived EF values and those obtained by 3DE in human subjects, 3DE provided better correlation than the biplane method (r = 0.94 versus r = 0.85). Real-time 3DE accurately measures left ventricular volumes transthoracically over a wide range of hemodynamic conditions in dogs and accurately determines EF in humans.

A SQUID based AC susceptometer for the investigation of large samples.

We developed a SQUID based susceptometer with a large available magnetized volume for the investigation of large objects. The magnetizing field is generated by a pair of Helmoltz coils. To achieve a high signal-to-noise ratio, the susceptometer is operated in a lock-in mode with an AC magnetizing field. A negative feedback control allows the rejection of the applied field with a relative residual of 1 x 10(-7). The apparatus was tested with substances of known magnetic susceptibility. The overall sensitivity, stated in terms of the magnetic moment, is better than 7 x 10(-11) A m2 for small samples.

The use of an inhomogeneous applied field improves the spatial sensitivity profile of an in vivo SQUID susceptometer.

We present a SQUID susceptometer with a non-homogeneous magnetizing field which is null at the sensing coil and increases towards the patient position with a constant gradient plus a cubic term at large distances. Compared with the magnetizing fields of similar instruments described in the literature, our gradient field enhances the signal due to internal organs with respect to the signal due to superficial tissue. Preliminary measurements have been performed on phantoms of known magnetic susceptibility. The advantage of using a non-homogeneous field compared with a uniform field has been investigated in the case of a double-layer phantom.

Detection and counting of specific cell populations by means of magnetic markers linked to monoclonal antibodies.

We report on experiments aimed at the assessment of a new method for cell marking. This method combines superparamagnetic particles, commonly used for cell separation, linked to monoclonal antibodies, and biomagnetic instrumentation featuring an extremely high magnetic field sensitivity. The final goal of the method is to locate and estimate specific cell populations in the human body. In this experiment, quantitative features of the method are evaluated in vitro with lymphocytes and carcinoma cells. Comparison between estimation and direct counting of cells is quite satisfactory and motivates further development of the technique.

Fetal magnetocardiographic mapping using independent component analysis.

Fetal magnetocardiography (fMCG) is the only noninvasive technique allowing effective assessment of fetal cardiac electrical activity during the prenatal period. The reconstruction of reliable magnetic field mapping associated with fetal heart activity would allow three-dimensional source localization. The efficiency of independent component analysis (ICA) in restoring reliable fetal traces from multichannel fMCG has already been demonstrated. In this paper, we describe a method of reconstructing a complete set of fetal signals hidden in multichannel fMCG preserving their correct spatial distribution, waveform, polarity and amplitude. Fetal independent components, retrieved with an ICA algorithm (FastICA), were interpolated (fICI method) using information gathered during FastICA iterations. The restored fetal signals were used to reconstruct accurate magnetic mapping for every millisecond during the average beat. The procedure was validated on fMCG recorded from the 22nd gestational week onward with a multichannel MCG system working in a shielded room. The interpolated traces were compared with those obtained with a standard technique, and the consistency of fetal mapping was checked evaluating source localizations relative to fetal echocardiographic information. Good magnetic field distributions during the P-QRS-T waves were attained with fICI for all gestational periods; their reliability was confirmed by three-dimensional source localizations.

Time course reconstruction of fetal cardiac signals from fMCG: independent component analysis versus adaptive maternal beat subtraction.

M-mode and pulsed Doppler echocardiography, cardiotocography and transabdominal fetal ECG are available in clinical practice to monitor fetal cardiac activity during advancing gestation, but none of these methods allows the direct measurement of morphological and temporal parameters for fetal rhythm assessment. Fetal magnetocardiograms (fMCGs) are noninvasive recordings of magnetic field variations associated with electrical activity of the fetal heart obtained with superconducting sensors positioned over the maternal abdomen inside a shielded room. Because of maternal cardiac activity, fMCGs are contaminated by maternal components that need to be eliminated to reconstruct fetal cardiac traces. The aim of the present work was to use two methods working in the time domain, an independent component analysis algorithm (FastICA) and an adaptive maternal beat subtraction technique (AMBS), for the retrieval of fetal cardiac signals from fMCGs. Detection rates of both methods were calculated, and FastICA and AMBS performances were compared in the context of clinical applications by estimating several temporal and morphological characteristics of the retrieved fetal traces, such as the shape and duration P-QRS-T waves, arrhythmic beat detection and classification, and noise reduction. Quantitative and qualitative comparison produced figures that always suggested that FastICA was superior to AMBS from the perspective of clinical use of the recovered fetal signals.

The study of steady magnetic fields associated with primary and secondary ST shift in ischaemic rabbit hearts.

The study of injury potentials associated with DC currents that generate the primary or secondary ST shifts during cardiac ischaemia is possible only through the invasive technique of the DC electrogram. Clinical surface ECG recordings are AC coupled and cannot be used. This paper reports the use of non-invasive and unshielded magnetocardiographic measurements to evaluate the DC injury currents associated with ST shifts during coronary artery occlusions in the isolated rabbit heart. The effect on the magnetic ST shift is studied under different ischaemic conditions including regional ischaemia, global ischaemia, global ischaemia following long periods of regional ischaemia, regional ischaemia after repeated episodes of reversible global ischaemia, and bilateral regional ischaemia. Recording of DC magnetic fields allows the characterization of primary and secondary ST displacement for each induced ischaemic condition. Our measurements show that the ST shift starts earlier when inducing ischaemia in hearts previously subjected to ischaemic episodes than in hearts where the ischaemia was produced for the first time.

[Percutaneous aortic valvuloplasty in the adult. When and why is now useful?]. / La valvuloplastica aortica percutanea nell'adulto. Quando e per chi è ancora utile?.

Percutaneous aortic valvuloplasty was introduced into clinical practice in 1986 and widely applied in elderly patients with symptomatic aortic stenosis. Nevertheless its results have been unsatisfactory over the mid to long term due to a high incidence of restenosis after 6-12 months. At the same time, patients over 70 years are more frequently undergoing surgical aortic valve replacement with low immediate postoperative mortality and good long term results. Although randomized trials are not available, aortic valve replacement seems to be a definitive therapeutic treatment when compared to the palliative result of aortic percutaneous valvuloplasty. However, since the complication rate of valvuloplasty carried out in cardiological centers with experienced personnel is low, this procedure is still indicated in selected patients. The very old (> 80 years) patients with associated systemic disease, and candidates for major surgery are referred for this procedure. Another indication for aortic valvuloplasty is severe aortic stenosis with cardiogenic shock; in this case, valve dilatation improves clinical status and acts as a "bridge" to surgery, enabling surgical intervention to be carried out at a later date. Nowadays, aortic percutaneous valvuloplasty is a possible alternative to surgical treatment in patients with an absolute surgical contraindication and in those who are in such poor clinical condition that they cannot be immediately referred to surgery. It is also useful for patients requiring urgent non-cardiac surgery (e.g., subjects with gastrointestinal bleeding). We discuss our results with this procedure which concord with those presented in the literature.

Carcinoid Heart Disease.

Carcinoid syndrome originates from metastatic carcinoid tumors localized in the gastrointestinal system, pancreas, biliary vessels, bronchi, ovaries, and testes; it is characterized by flushing, telangiectasias, diarrhea, bronchoconstriction, and fibrous endocardial plaques in the heart. Cardiac involvement is detected by echocardiography in over 50% of patients with this syndrome. Right-sided valvular heart disease occurs frequently in patients with carcinoid syndrome, involving most commonly the tricuspid and pulmonary valves. Involvement of the left-sided valves rarely occurs. Medical therapy for carcinoid heart disease includes digitalis and diuretics for congestive heart failure symptoms; the effects of carcinoid syndrome can be treated with the use of somatostatin analogues. Conventional chemotherapy has been of little beneficial value, with response rates of only 10% to 30%. The use of octreotide, a long-acting and potent somatostatin analogue, is a major advance in the management of carcinoid tumors. In addition to providing effective symptom relief in malignant carcinoid syndrome, octreotide can also be used for diagnostic purposes. Despite its expense, octreotide is the current agent of choice for the treatment of this condition. Analogues with different receptor specificities and pharmacokinetics hold promise for the future. Valve surgery is the only definitive treatment for intractable right-sided heart failure. Although cardiac surgery carries high perioperative mortality, marked symptomatic improvement occurs in survivors. Surgical intervention therefore should be considered in the appropriate patients when cardiac symptoms become severe. Balloon valvulotomy in patients with severe pulmonary artery stenosis often results in palliation of symptoms; however, surgery still is required often in these patients.

Biological effects of electromagnetic fields.

Nowadays, concerns about hazards from electromagnetic fields represent an alarming source for human lives in technologically developed countries. We are surrounded by electromagnetic fields everywhere we spend our working hours, rest or recreational activities. The aim of this review is to summarize the biological effects due to these fields arising from power and transmission lines, electrical cable splices, electronic devices inside our homes and work-places, distribution networks and associated devices such as cellular telephones and wireless communication tower, etc. Special care has been reserved to study the biological effects of electromagnetic fields on cell lines of the mammalian immune system about which our research group has been working for several years.

[Spectral properties of perselenosulfide]. / Proprieta' spettrali del perselenosolfuro.

The riductive scission of a diselenide (selenocystamine) produced by disulfide gives the perselenosulfide, a new compound. Its formation kinetics, carried out a several pH values, were recorded spectrophotometrically at 375 nm. The lability of perselenosulfide bond by cianolysis has been demonstrated.

On the interpretation of the early part of the liver time-activity curve: double tracer experiment.

A double tracer experiment was performed in one normal subject and in one cirrhotic patient to better understand the tracer kinetics in the liver area during the first minute after injection. From a comparison of the time-activity curves obtained in each subject by subsequent iv administration of a nonextracted and an extracted (colloidal) tracer it was possible to distinguish the contribution due to the transport mechanism through intrahepatic and extrahepatic circulation and that due to reticuloendothelial extraction. Three main phases were identified on the curves and an attempt was made to interpret their significance. Results of the study may be of interest for the development of models which could possibly provide more reliable noninvasive quantitation of liver hemodynamics and reticuloendothelial function with the use of one or both types of tracer.