Unggul P. Juswono

Oscillations in proton transport revealed from simultaneous measurements of net current and net proton fluxes from isolated root protoplasts: MIFE meets patch-clamp

Proton fluxes were measured non-invasively on patch-clamped protoplasts isolated from wheat roots using an external H + electrode to measure the electrochemical gradient in the external solution. Under voltage clamp in the whole-cell configuration, the H + fluxes across the plasma membrane could be measured as a function of voltage and time and correlated with the simultaneous measurements of membrane current. Protoplasts could exist in three states based on the current–voltage (I–V) curves and the flux–V curves. In the pump-state where the membrane voltage (Vm) was more negative than the electrochemical equilibrium potential for potassium (E K ), a net efflux of H + occurred that was voltage-dependent such that the efflux increased as Vm was clamped more positive. In the K-state, where Vm was close to E K , similar flux–V curves were observed. In the depolarised state where Vm was greater than E K the proton flux was characterised by a net influx of H + (H + -influx state) that reversed direction at more positive values of Vm. The inhibitory effect of DCCD and stimulatory effect of fusicoccin were used to correlate current and H + flux through the H + -ATPase for which there was reasonably good agreement within the limits of the flux measurements. Some protoplasts were kept in the whole-cell configuration for up to 3 h revealing slow sustained oscillations (period about 40 min) in H + flux that were in phase with oscillations in free-running Vm. These oscillations were also observed under voltage clamp, with membrane current in phase with H + flux, but which became damped out after a few cycles. The oscillations encompassed the pump-state, K + -state and H + -influx-state. The H +- flux–V curves and I–V curves were used to model the electrical characteristics of the plasma membrane with H + -ATPase, inward and outward K + rectifiers, a linear conductance, and a passive H + influx possibly through gated proton channels.

Protoplast ion fluxes: their measurement and variation with time, position and osmoticum

Abstract. The ability to measure directly individual protoplast ion fluxes is a valuable addition to patch clamp and other techniques when using protoplasts to study membrane transporters. Before interpreting observations on protoplasts in terms of behaviour of intact cells and tissues, some methodological questions should be addressed. These include effects of space and time variations of transporter activities over the membrane, the osmotic dependence of specific ion transporters and the effect of the regenerating cell wall. In this study net H+ and Ca2+ fluxes were measured from individual corn (Zea mays L.) coleoptile protoplasts using a non-invasive microelectrode technique for ion flux measurements. For Ca2+, the flux distribution was almost symmetrical, ranging ±30 nmol · m−2 · s−1 around zero. For H+ it was skewed towards efflux ranging from −100 to +10 nmol · m−2 · s−1. The distribution of H+ fluxes through the protoplast surface was a complex mosaic which changed with time, sometimes showing oscillations. These flux variations with time and position around the surface, apparently driven by endogenous mechanisms, may be relevant to protoplast pH homeostasis. When the new cell wall was partially regenerated on the next day, the correlation between H+ and Ca2+ fluxes increased, which is consistent with the weak-acid Donnan-Manning model of cell wall ion exchange.

A study of the correlation between ultrafine particle emissions in motorcycle smoke and mice erythrocyte damages

Sharply increasing of motor vehicles every year contributes to amounts of ultrafine particles (UFPs) in the air. Besides, the existence of UFPs in the blood may cause erythrocyte damages that subject to shape deformation. This study was aimed to investigate the influence of UFPs in the motorcycle smoke exposed to mice in different concentrations to the erythrocyte damages. The experiments were conducted by injecting the motorcycle smoke with the varied amounts in an experimental chamber (dimension of 30×20×20cm3) where the mice were put in advance for exposuring twice a day (100s). Total numbers of UFPs in the smoke were calculated by measuring the total concentrations multiplied by the smoke debit. They were measured using a TSI 8525 P-Trak UPC. The effects of the smoke exposures in the mices erythrocytes related to the UFPs in the smoke were observed by a binocular CX-31 Computer Microscope after the 2nd, 4th, 6th, 8th, and 10th exposure days. The erythrocyte damages were calculated from the total abnormal erythrocytes divided by the total erythrocytes. Our results showed that more UFPs exposed to mice resulted in more the erythrocytes damages. Longer exposures caused more damages of the mice erythrocytes. This study found significant correlations between the numbers of UFPs exposed to mice and the erythrocyte damages. Our finding gives important evidence that motorcycle emissions especially UFPs affect on health.

An observation of histological evidence on internal organ damages in mice caused by repeated exposures to motorcycle emissions

Motor vehicle emissions have been identified as a source of ultrafine particles, which have significant impacts on human health. Repeated and prolonged exposure to ultrafine particles may have a significant association with organ damage. Here, we evaluated the correlation between repeated exposure to ultrafine particles and organ damage in mice. Motorcycle emissions were injected into an exposure chamber with mice for a period of 20 seconds. This treatment was conducted over 10 days. The mice were sacrificed on the 2nd, 4th, 6th, 8th, and 10th days for organ preparations. Based on the results, motorcycle emission exposure caused organ damage in mice, with different severities depending on the organ. The highest damage was found for the lung, followed by the kidney, erythrocytes, and liver.

Measurements of PM 2.5 motor emission concentrations and the lung damages from the exposure mice

Motor cycles grows very fast from year to year in the developing countries, especially in Indonesia. On the other sides, the emissions contribute to high pollutants in the air. In terms of PM2.5 emissions affect on human health. However the data of PM2.5 emitted by motorcycles are unavailable, and the relationship between PM2.5 containing in the motor smokes and the impacts on human health is also unknown. This study was aimed to investigate the impacts of PM2.5 in the motor cycle smokes on mice/s lung damages by conducting measurements of varied PM2.5 concentrations exposed to mice and observing the mice lung damages. The measurements of PM2.5 concentrations in the smokes were conducted by introducing motor cycle smokes in a chamber with the dimension of 30×20×20 cm3 with a pump with the varies of time. The PM2.5 concentrations were measured using a Kanomax 3443 Digital Dust Monitor. Investigation of the impacts PM2.5 on mice lung were carried out by placing mice in the chamber for smoke exposures with the varied concentrations of PM2.5 in the smokes for 100 s twice a day. The measurements of the mice lung damages were carried out by observing the normal and abnormal alveolus using a Binocular CX-31 Computer Microscope with 400x magnification after the mice experienced smoke exposures for two days. The results showed that longer time of introducing smokes in the chamber resulted in higher concentrations of PM2.5. The measured concentrations are found varied between 1.2 and 19.3 mg/m3 depending on the operating time and the motor sample. Higher concentration of PM2.5 in the smokes exposed to the mice caused more damages of the mice lung. The correlation is obtained in the second order polynomial function with the R2 > 0.9.

Varied dose exposures to ultrafine particles in the motorcycle smoke cause kidney cell damages in male mice

Graphical abstract

Analysis of Voltage Difference Characteristics of Albumin Membranes and Vitelin Membranes of Chicken Eggs through Diazinon Effects

Cells are the most important structural and functional part of any organism. Cells are protected by cell membranes that envelop cells, and separate cells with their environment. The effectiveness of the work of a cell membrane can be disrupted for various reasons, one of which is pesticide contamination. Pesticides in cells can damage cell metabolism or cell membrane permeability. Therefore, this study aims to determine how big the effect of diazinon insecticides on the characteristics of different potential membrane vitelin and membrane albumin chicken egg cells. Measurements were made using the microelectrode connected with the picoscope 5000 series, then the data appears displayed through the PC. The results of membrane potential measurements showed that the greater the concentration of insecticide given, the potential value of chicken egg membrane increased. Potential Differential Values Membrane of eggs with different concentrations ranged in -145 mV - 249 mV on vitelin membrane, and -142,5 mV - 242,5 mV on albumin membrane

Pengaruh Radiasi Gamma dan Ekstrak Temulawak (Curcuma xanthorrhiza) terhadap Kadar SGPT Hepar Mencit (Mus musculus)

Gamma radiation can causes cells damage, because the interaction between radiations with cells can produce free radical. The compound that can neutralize and catch free radical is called antioxidant. The objectives of this research were to analyze influence of gamma radiation and Curcuma xanthorrhiza extract to the SGPT level to the mice’s liver. This research uses eighty mices and they were divided into three group, negative control (K-), radiation non extract (R-) and extract plus radiation (R+). The doses of treatment of Curcuma xanthorrhiza extract are 1,4 gr/kg body weight; 2,0 gr/kg body weight; 2,6 gr/kg body weight; 3,2 gr/kg body weight and 3,8 gr/kg body weight, while the exposure of gamma radiation are given during 10 minutes, 20 minutes, 30 minutes, 40 minutes and 50 minutes. The measuring of SGPT level in mice’s serum is done by using blood analyzer. The results showed that free radical can damages liver cells and breaks cell wall, so SGPT level in blood to be high. In a row of increasing of Curcuma xanthorrhiza extract dose, then SGPT level to be low.

Estimation of Radiation Dose Calculation on Non Static Organs

In this study developed a program calculating the estimated radiation dose to the cancer by using a program that can run on a variety of computer operating systems platforms. The program used is a Java application programming interface (API) Greenfoot. The program determines the value of the accumulated dose received by each pixel in multiple beam irradiation with attenuation values obtained from the analysis of data Hounsfield value CT-scan . Hounsfield value of each pixel is generated based on the relationship value of grayscale linearity then. Hounsfield value of the CT-scan converted into the value of the gamma-ray attenuation. The dose received by the skin surface used as a reference to determine the length of time considered organ irradiation in static conditions on the program. Movement of the target irradiation in treatment may reduce the efficiency level of success. Moving organ radiation Treatment can be done. Radiation pulse period set for the radiation beam on the target in this phase organ reaches ± 60% of the maximum volume. A pulse rays will be effect the long exposure time can be calculated with the program.

Effects of Visible Spectrum on Potential Differential Response to Soybean Sprouts (Glycine max (L.) Merill) during photosynthesis

The absorption visible light spectrum by chlorophyll cause excited electron. The transition of electron in photosynthesis process result electrical potential difference response which can be utilized to spur metabolism process in the plants itself. This research used the seedling of Glycine max (L.) and the varietas is sindoro. The measurement of the potential difference response is done by adding of visible light (red, yellow, green, blue and violet) alternately to the bean sprouts of Glycine max (L.) Merill . The light distribution is being done for 1 second, 5 second and constantly. The result of the research showed the effect of the visible light distribution constantly in the photosynthesis process. In the red light occurs maximum potential difference response. In peak I resulted -147,3 mV and peak II resulted -137 mV. The minimum potential difference response is resulted by green light. in peak I resulted -68,8 mV and peak II resulted -56,75 mV. The effect of the visible light distribution during any time in the photosynthesis process. In the red light occurs maximum potential difference response. In peak I resulted -119,2 mV for 1 second and -146 Mv for 5 second. In peak II resulted - 112,3 mV for 1 second and -125,5 mV for 5 second. The minimum potential difference response is resulted by green light. In peak I resulted -64,75 mV for 1 second, -69,5 mV for 5 second, and in peak II resulted -55,9 mV for 1 second and -56 mV for 5 second.