Svetlozar Velizarov

The effects of radiofrequency fields on cell proliferation are non-thermal.

The number of reports on the effects induced by radiofrequency (RF) electromagnetic fields and microwave (MW) radiation in various cellular systems is still increasing. Until now no satisfactory mechanism has been proposed to explain the biological effects of these fields. One of the current theories is that heat generation by RF/MW is the cause, in spite of the fact that a great number of studies under isothermal conditions have reported significant cellular changes after exposure to RF/MW. Therefore, this study was undertaken to investigate which effect MW radiation from these fields in combination with a significant change of temperature could have on cell proliferation. The experiments were performed on the same cell line, and with the same exposure system as in a previous work [S. Kwee, P. Raskmark, Changes in cell proliferation due to environmental non-ionizing radiation: 2. Microwave radiation, Bioelectrochem. Bioenerg., 44 (1998), pp. 251-255]. The field was generated by signal simulation of the Global System for Mobile communications (GSM) of 960 MHz. Cell cultures, growing in microtiter plates, were exposed in a specially constructed chamber, a Transverse Electromagnetic (TEM) cell. The Specific Absorption Rate (SAR) value for each cell well was calculated for this exposure system. However, in this study the cells were exposed to the field at a higher or lower temperature than the temperature in the field-free incubator i.e., the temperature in the TEM cell was either 39 or 35 +/- 0.1 degrees C. The corresponding sham experiments were performed under exactly the same experimental conditions. The results showed that there was a significant change in cell proliferation in the exposed cells in comparison to the non-exposed (control) cells at both temperatures. On the other hand, no significant change in proliferation rate was found in the sham-exposed cells at both temperatures. This shows that biological effects due to RF/MW cannot be attributed only to a change of temperature. Since the RF/MW induced changes were of the same order of magnitude at both temperatures and also comparable to our previous results under isothermal conditions at 37 degrees C, cellular stress caused by electromagnetic fields could initiate the changes in cell cycle reaction rates. It is widely accepted that certain classes of heat-shock proteins are involved in these stress reactions.

CHANGES IN CELLULAR PROTEINS DUE TO ENVIRONMENTAL NON-IONIZING RADIATION. I. HEAT-SHOCK PROTEINS

This paper describes the effect of weak microwave fields on the amounts of heat-shock proteins in cell cultures at various temperatures. The field was generated by signal simulation of the Global System for Mobile communications (GSM) of 960 Mhz, used in portable phones. Transformed human epithelial amnion (AMA) cells, growing on glass coverslips, were exposed in a transverse electromagnetic (TEM) cell to a microwave field, generating a specific absorption rate (SAR) of 2.1 mW.kg−1 in the cells. Exposure temperatures were 35, 37, and 40 ± 0.1°C, respectively, and the exposure time was 20 min. The heat-shock proteins Hsp-70 and Hsp-27 were detected by immuno-fluorescence. Higher amounts of Hsp-70 were present in the cells exposed at 35 and 37°C than in the sham-exposed cells. These effects can be considered to be athermal, since the field strength was much lower than the safety standard for absence of heat generation by microwave fields. There was no significant response in the case of Hsp-27.

Biotransformation of glucose to free gluconic acid by Gluconobacter oxydans: substrate and product inhibition situations

Abstract Microbial oxidation of glucose to free gluconic acid by Gluconobacter oxydans batch cultures was studied with a special reference to substrate and product inhibition. Process kinetics were evaluated by comparing different inhibition models. Substrate inhibition can be best described by the equation of Tseng and Wayman predicting linear inhibition of growth above a characteristic threshold glucose concentration. Non-linear inhibition kinetics was observed, however, for product inhibition and the model of Levenspiel was found to fit the experimental data. The biokinetic model parameters, productivities and conversion degrees on media with different glucose and gluconic acid concentrations were calculated and compared regarding the principal limits of process efficiency. Probable mechanisms of inhibition and possible techniques for their minimization are also discussed.

Kinetics of nitrate and perchlorate removal and biofilm stratification in an ion exchange membrane bioreactor

The biological degradation of nitrate and perchlorate was investigated in an ion exchange membrane bioreactor (IEMB) using a mixed anoxic microbial culture and ethanol as the carbon source. In this process, a membrane-supported biofilm reduces nitrate and perchlorate delivered through an anion exchange membrane from a polluted water stream, containing 60 mg/L of NO₃⁻ and 100 μg/L of ClO₄⁻. Under ammonia limiting conditions, the perchlorate reduction rate decreased by 10%, whereas the nitrate reduction rate was unaffected. Though nitrate and perchlorate accumulated in the bioreactor, their concentrations in the treated water (2.8 ± 0.5 mg/L of NO₃⁻ and 7.0 ± 0.8 μg/L of ClO₄⁻, respectively) were always below the drinking water regulatory levels, due to Donnan dialysis control of the ionic transport in the system. Kinetic parameters determined for the mixed microbial culture in suspension showed that the nitrate reduction rate was 35 times higher than the maximum perchlorate reduction rate. It was found that perchlorate reduction was inhibited by nitrate, since after nitrate depletion perchlorate reduction rate increased by 77%. The biofilm developed in the IEMB was cryosectioned and the microbial population was analyzed by fluorescence in situ hybridization (FISH). The results obtained seem to indicate that the kinetic advantage of nitrate reduction favored accumulation of denitrifiers near the membrane, whereas per(chlorate) reducing bacteria were mainly positioned at the biofilm outer surface, contacting the biomedium. As a consequence of the biofilm stratification, the reduction of perchlorate and nitrate occur sequentially in space allowing for the removal of both ions in the IEMB.

Mercury removal from water streams through the ion exchange membrane bioreactor concept

Mercury is a highly toxic heavy metal that causes human health problems and environmental contamination. In this study, an ion exchange membrane bioreactor (IEMB) process was developed to achieve Hg(II) removal from drinking water and industrial effluents. Hg(II) transport through a cation exchange membrane was coupled with its bioreduction to Hg(0) in order to achieve Hg removal from concentrated streams, with minimal production of contaminated by-products observed. This study involves (1) membrane selection, (2) demonstration of process effectiveness for removing Hg from drinking water to below the 1ppb recommended limit, and (3) process application for treatment of concentrated water streams, where >98% of the Hg was removed, and the throughput of contaminated water was optimised through membrane pre-treatment. The IEMB process represents a novel mercury treatment technology with minimal generation of contaminated waste, thereby reducing the overall environmental impact of the process.

Removal of trace mono-valent inorganic pollutants in an ion exchange membrane bioreactor: analysis of transport rate in a denitrification process

The ion exchange membrane bioreactor (IEMB) uses a mono-anion permselective membrane as a barrier between a water stream, containing a target polluting anion, and a biocompartment, containing a suitable driving counter-ion for coupled counter-diffusion of the target pollutant and a microbial culture capable of its bioreduction to harmless product(s). A resistances-in-series trace counter-ion approximation model, based on the Fick formalism and the Donnan equilibrium principle is proposed for analysis of the transport rate of dilute inorganic anionic pollutants with relevance to drinking water treatment. Transport of co-ions (cations) is negligible due to their electrostatic repulsation (Donnan exclusion) from the positively charged membrane. The model is tested for a system, containing nitrate as the target pollutant and chloride as the driving bulk counter-ion. The effect of the most important process variables on the target pollutant flux are assessed using the model, making possible the comparison between different IEMB process options, which can be quantified and used for process design. The impact of the bioconversion reaction on the mass transfer resistance to the target pollutant transport is also discussed.

Nitrate removal in a closed marine system through the ion exchange membrane bioreactor

The accumulation of nitrate in closed marine systems presents a problem for both the marine life and the environment. The present study, proposes the application of the ion exchange membrane bioreactor (IEMB) concept for removing nitrate from marine systems, such as aquaculture tanks or marine aquariums. The results obtained demonstrate that the IEMB was able to remove naturally accumulated nitrate from water taken from a public marine aquarium (Oceanário de Lisboa) and bioconvert it, in an isolated compartment (biocompartment), to molecular nitrogen, thus preventing secondary contamination of the treated water by microbial cells, metabolic by-products and excess of carbon source (ethanol). This system allowed for the removal of nitrate at concentrations of 251 and 380 mg/l down to below 27 mg/l exchanging it for chloride. Under the studied operating conditions, the IEMB proves to be a selective nitrate removing technology preserving the initial water composition with respect to cations, due to the Donnan exclusion effect from the membrane, and minimizing the counter diffusion of anions other than nitrate and chloride, due to the use of water with the same ionic composition in the biocompartment. This is an advantage of the IEMB concept, since the quality of the water produced would allow for the reutilisation of the treated water in the aquarium, thereby reducing both the wastewater volume and the use of fresh water.

Mechanism of charged pollutants removal in an ion exchange membrane bioreactor : Drinking water denitrification

The mechanism of anionic pollutant removal in an ion exchange membrane bioreactor (IEMB) was studied for drinking water denitrification. This hybrid process combines continuous ion exchange transport (Donnan dialysis) of nitrate and its simultaneous bioreduction to gaseous nitrogen. A nonporous mono-anion permselective membrane precludes direct contact between the polluted water and the denitrifying culture and prevents secondary pollution of the treated water with dissolved nutrients and metabolic products. Complete denitrification may be achieved without accumulation of NO3(-) and NO2(-) ions in the biocompartment. Focus was given to the effect of the concentration of co-ions, counterions, and ethanol on the IEMB performance. The nitrate overall mass transfer coefficient in this hybrid process was found to be 2.8 times higher compared to that in a pure Donnan dialysis process without denitrification. Furthermore, by adjusting the ratio of co-ions between the biocompartment and the polluted water compartment, the magnitude and direction of each individual anion flux can be easily regulated, allowing for flexible process operation and control. Synthetic groundwater containing 135-350 mg NO3(-) L(-1) was treated in the IEMB system. A surface denitrification rate of 33 g NO3(-) per square meter of membrane per day was obtained at a nitrate loading rate of 360 g NO3(-) m(-3)d(-1), resulting in a nitrate removal efficiency of 85%.

The effect of carbon source on the biological reduction of ionic mercury

Mercury (Hg) is the most highly toxic heavy metal, and must be removed from waterways to very low levels. Biologically mediated mercury removal is an emerging technology that has the potential to be robust, efficient and cost-effective. In this study, the impact of carbon source on the behaviour and microbial community composition of mixed microbial cultures was evaluated, and their performance was compared with a pure culture of Pseudomonas putida spi3. Glucose and acetate, two carbon sources that are commonly present in wastewaters, were chosen for this study. Distinct microbial populations were enriched with each carbon source. Glucose led to a more suitable microbial culture for Hg(2+) bioreduction that was able to reduce Hg(2+) at faster rates when compared to acetate. Furthermore, acetate consistently led to poorer process performance, irrespective of the microbial culture, possibly due to the formation of mercuric acetate complexes. It is proposed that glucose can be a more beneficial carbon source than acetate for the successful operation of Hg bioremediation systems.

Ion exchange membrane bioreactor for selective removal of nitrate from drinking water: control of ion fluxes and process performance.

An ion exchange membrane bioreactor (IEMB), consisting of a monoanion permselective membrane dialyzer coupled to a stirred anoxic vessel with an enriched mixed denitrifying culture, has been studied for nitrate removal from drinking water. The influence of nitrate and chloride concentrations on the selectivity of nitrate transport in the IEMB process was investigated. With appropriate dosing of chloride ions to the IEMB biocompartment, it was possible to regulate the net bicarbonate flux in the system, thus maintaining the bicarbonate concentration in the treated water at the desired level. The latter was not possible to achieve in Donnan dialysis, operated as a single process in which, besides the lower nitrate removal efficiency found, bicarbonate was co‐extracted together with nitrate from the polluted water stream. Residual carbon source (ethanol) and nitrite were not detected in the treated water produced in the IEMB system. With a concentration of nitrate in the polluted water three times higher than the maximum contaminant level of 50 mg L−1 allowed, the IEMB process was successfully operated for a period of 1 month before exceeding this limit.