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General Physiology and Biophysics

Volume 34, 2015, No. 1


  Editorial Commentary: The meth brain: methamphetamines alter brain functions via NMDA receptors
Juliane Proft 1), Norbert Weiss

1)Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic.

Commentary to: Functional changes in pyramidal neurons in the chronic methamphetamine-treated rat. (Gen. Physiol. Biophys. 2015, pp.5-12).

General Physiology and Biophysics. Volume 34, 2015, No. 1: 1-3.

  Functional changes in piriform cortex pyramidal neurons in the chronic methamphetamine-treated rat
Nobuaki Hori 1), Tomoko Kadota, Norio Akaike

1)Research Division for Life Sciences, Kumamoto Health Science University, Izumi 325, Kita-ku, Kumamoto City, Kumamoto 861-5598, Japan.

Chronic treatment of rats with methamphetamine (MAP) causes a range of functional changes to the central nervous system (CNS), including a toxicity that is widespread throughout the brain (Frost and Cadet 2000; Fasihpour et al. 2013). In this report, we examined the effect of chronic MAP treatment on pyramidal neurons of the rat piriform cortex, an area involved in sensory processing, associative learning and a model system for studies on synaptic plasticity. MAP treatment significantly depolarized the membrane potential and decreased neuronal input resistance. Furthermore, the voltage-dependence of both AMPA and NMDA responses was disturbed by chronic MAP treatment, and the extent of long-term potentiation (LTP) was decreased. Morphological changes of MAP-treated rat pyramidal neurons were observed as blebbing of the dendrite trees. The changes we observed represent detrimental effects on the function of piriform cortical neurons further illustrating deficits in synaptic plasticity extend beyond the hippocampus. These changes may contribute to behavioural deficits in chronic MAP-treated animals.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 5-12.

  Enhancement of cytotoxic effect on human head and neck cancer cells by combination of photodynamic therapy and sulforaphane
Sang Lee 1), Hee-Jun Hwang, Jang-In Shin, Jin-Chul Ahn, Phil-Sang Chung

1)Department of Otolaryngology-Head and Neck Surgery, College of Medicine, Dankook University, Cheonan, Korea.

Photodynamic therapy (PDT) is a method to treat cancers using photosensitizer and light. PDT has been tried for several tumors. However, the clinical applications are limited by the toxicity of photosensitizer and narrow effect. Sulforaphane (SFN) is a material of isothiocyanate group and known to have anticancer effect. We evaluated the cytotoxic effect of PDT combined with SFN on human head and neck cancer cells. We measured the cell viability, extent of apoptosis and necrosis, reactive oxygen species (ROS) generation and caspase activation. Cell viability was decreased significantly by combination treatment. Cellular apoptosis and necrosis were increased in combination treatment compared to SFN or PDT. ROS generation was also higher in combination treatment than single treatment. In combination treatment group, apoptosis and necrosis were decreased by administration of sodium azide (SA) which is scavenger of ROS. Increased caspase activation in combination treatment was also inhibited by SA. Combination of PDT and SFN led to enhanced cytotoxic effect on head and neck cancer cells. Combination treatment promoted the ROS generation, which induced cell death through activation of caspase pathway.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 13-21.

  Does static magnetic field-exposure induced oxidative stress and apoptosis in rat kidney and muscle? Effect of vitamin E and selenium supplementations
Soumaya Ghodbane 1), Aida Lahbib, Mohamed Ammari, Mohsen Sakly, Hafedh Abdelmelek

1)Laboratoire de Physiologie Intégrée, Faculté des Sciences de Bizerte, 7021 Jarzouna, Tunisia.

Static magnetic fields (SMFs) effect observed with radical pair recombination is one of the well-known mechanisms by which SMFs interact with biological systems. Our aim was to study whether SMF induces oxidative stress and apoptosis in rat tissues and to evaluate the possible protector effect of selenium (Se) and vitamin E (vit E) supplementations. Rats were randomly divided into control, SMF-exposed, Se-treated, vit E-treated, SMF exposed rats and co-treated with Se, and SMF exposed rats and co-treated with vit E. After animal sacrifice, catalase (CAT) activity and malondialdehyde (MDA) concentration were measured and apoptosis inducing factor (AIF) immunohistochemical labeling was performed in kidney and muscle. Exposure of rats to SMF (128 mT, 1 h/day for 5 days) increased the MDA concentrations (+25%) and CAT activities (+34%) in kidney but not in muscle. By contrast, the same treatment failed to induce a caspase-independent pathway apoptosis in both tissues. Interestingly, Se pre-treatment inhibited the increase of MDA concentrations and CAT activities in kidney in SMF-exposed rats. However, vit E administration corrected only MDA levels in rat kidney. In conclusion, exposure to SMF induced oxidative stress in kidney that can be prevented by treatment with Se or vit E.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 23-32.

  Differential impact of bortezomib on HL-60 and K562 cells
Katarína Kliková 1), Andrea Štefaniková, Ivana Pilchová, Jozef Hatok, Peter Chudý, Juraj Chudej, Dušan Dobrota, Peter Racay

1)Department of Medical Biochemistry, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Martin, Slovak Republic.

Bortezomib (PS-341, or Velcade), reversible inhibitor of 20S proteasome approved for the treatment of multiple myeloma and mantle cell lymphoma, exhibited a cytotoxic effect toward other malignancies including leukaemia. In this study, we have documented that incubation of both HL-60 and K562 leukaemia cells with nanomolar concentrations of bortezomib is associated with the death of HL-60 cells observed within 24 hours of incubation with bortezomib and the death of K562 cells that were observed after 72 hours of incubation with bortezomib. The relative resistance of K562 cells to bortezomib correlated well with significantly higher expression of HSP27, HSP70, HSP90α, HSP90β and GRP75 in these cells. Incubation of both HL-60 and K562 cells with bortezomib induced a cleavage of HSP90β as well as expression of HSP70 and HSP90β but bortezomib did not affect levels of HSP27, HSP90α, GRP75 and GRP78. The death of both types of cells was accompanied with proteolytic activation of caspase 3 that was observed in HL-60 cells and proteolytic degradation of procaspase 3 in K562 cells. Our study has also pointed to essential role of caspase 8 in bortezomib-induced cleavage of HSP90β in both HL-60 and K562 cells. Finally, we have shown that bortezomib induced activation of caspase 9/caspase 3 axis in HL-60 cells, while the mechanism of death of K562 cells remains unknown.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 33-42.

  Role of resveratrol on the cytotoxic effects and DNA damages of iododeoxyuridine and megavoltage radiation in spheroid culture of U87MG glioblastoma cell line
Fariba Firouzi 1), Samideh Khoei, Hamid Mirzaei

1)Radiology Technology Department, School of Paramedicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

The purpose of this study was to evaluate the effect of resveratrol on cytogenetic damages of iododeoxyuridine (IUdR) and x-ray megavoltage radiation (6 MV) in spheroid model of U87MG glioblastoma cancer cell line using clonogenic and alkaline comet assay. Cells were cultured as spheroids (350 µm) that were treated with 20 μM resveratrol, 1 μM IUdR and 2 Gy of 6 MV x-ray. After treatment, viability of the cells, colony forming ability and the induced DNA damages were examined using trypan blue dye exclusion, colonogenic and alkaline comet assay, respectively. Our results showed that resveratrol could significantly reduce the colony number and induce the DNA damages of the cells treated with IUdR in combination with 6 MV x-ray radiation. That results indicated that resveratrol as an inhibitor of hypoxia inducible factor 1 alpha (HIF-1α) protein in combination with IUdR as a radiosensitizer enhanced the radiosensitization of glioblastoma spheroid cells.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 43-50.

  Protection or cytotoxicity mediated by a novel quinonoid-polyphenol compound?
Ivana Milackova 1), Lucia Rackova, Magdalena Majekova, Natasa Mrvova, Milan Stefek

1)Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences, Dubravska cesta 9, 841 04 Bratislava, Slovak Republic.

Many natural and synthetic quinones and naphthoquinones possess a variety of beneficial pharmacological properties. In plants, the cytotoxic properties of quinones serve in their defensive roles against invading bacteria, fungi and parasites. In this regard many quinones as well as polyphenols, exerting generally toxicity at high dosages, are able to induce favorable hormetic responses at a low dosage. The novel chloronaphthoquinone derivative of quercetin (CHNQ) showed a profound cytotoxicity followed by enhancement of intracellular generation of oxidants in human neonatal B-HNF-3 fibroblasts. Its synthetic precursors, quercetin and 2-chloro-3-hydroxy-[1,4]naphthoquinone, failed to induce these effects, and paradoxically, only CHNQ at a low concetration provided partial protection of the cells against oxidative challenge. Thus, the novel quinonoid-polyphenol CHNQ might have a merit in the search for new prospective agents in prevention and management of ageing and ageing-related pathologies.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 51-64.

  Thymoquinone supplementation reverses lead-induced oxidative stress in adult rat testes
Aymen Mabrouk 1), Hassen Ben Cheikh

1)Laboratory of Histology and Cytogenetic (Research unit of Genetic, Genotoxicity and Child Disease UR12ES10), Faculty of Medicine, 5019 Monastir, Tunisia.

The purpose of the present study was to investigate the potential protective effect of thymoquinone (TQ), the major active ingredient of volatile oil of Nigella sativa seeds, against Pb-induced testicular oxidative stress. Adult male rats were randomized into four groups: control group which received no treatment, Pb group was exposed to 2000 ppm Pb acetate in drinking water, Pb-TQ group was co-treated with Pb plus TQ (5 mg/kg b.w./day, p.o.) and TQ group receiving only TQ (5 mg/kg b.w./day, p.o.). All treatments were applied for 5 weeks. Pb treatment induced oxidative stress status in testes as evidenced by a significant decrease in the antioxidant enzymes activities such as superoxide dismutase, glutathione peroxidase and catalase, and in the reduced glutathione content and in a significant increase in the level of malondialdehyde. Interestingly, TQ supplementation completely reversed these biochemical changes caused by Pb to the control values. In conclusion, our results suggest, for the first time, that TQ is very efficient in preventing Pb-induced testicular oxidative stress. This study will open new perspectives for the clinical use of TQ in Pb intoxication.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 65-72.

  Influence of oak wood polyphenols on cysteine, homocysteine and glutathione total levels and PON1 activities in human adult volunteers - a pilot study
Zuzana Deáková 1), Zuzana Országhová, Lucia Andrezálová, Peter Slezák, Jozef Lehotay, Jana Muchová, Carolina Bürki, Zdeňka Ďuračková

1)Institute of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, Comenius University, Sasinkova 2, 811 08 Bratislava, Slovak Republic.

Oxidative stress reflects an imbalance between antioxidants and pro-oxidants. Many diseases like atherosclerosis or heart failure are involved in oxidative stress. Increased oxidative stress is one of the potential contributing factors to aging. The aim of this study was to monitor the total thiol levels as markers of oxidative stress in 20 healthy volunteers after polyphenols intake (extract from the French oak wood Quercus robur – Robuvit® (300 mg/day)). Polyphenols are known as biomodulators with antioxidant activities. Homocysteine, cysteine and glutathione total levels were determined by using HPLC with electrochemical detection. The activity of the antioxidant enzyme paraoxonase-1 toward two substrates was determined by spectrophotometry. The level of thiol compounds and paraoxonase-1 activities were controlled after run-in (week 0), intervention (week 4) and washout (week 6) period. After the intervention period the results showed that Robuvit® had no significant influence on glutathione level (p = 0.382) and paraoxonase activities towards both, arylester and lactone substrates. On the other hand, homocysteine and cysteine levels decreased significantly (p = 0.029; p < 0.001, respectively). The negative correlation between paraoxonase lactonase activity and homocysteine level was noticed. This confirms that paraoxonase might play an important role in homocysteine-thiolactone metabolism.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 73-80.

  The effect of chronic peripheral nesfatin-1 application on blood pressure in normal and chronic restraint stressed rats: related with circulating level of blood pressure regulators
Ceylan Ayada 1), Günfer Turgut, Sebahat Turgut, Zuhal Güçlü

1)Department of Physiology, Faculty of Medicine, University of Dumlupınar, Kütahya, Turkey.

Nesfatin is a peptide secreted by peripheral tissues, central and peripheral nervous system. It is involved in the regulation of homeostasis. Although the effects of nesfatin-1 on nutrition have been studied widely in the literature, the mechanisms of nesfatin-1 action and also relations with other physiological parameters are still not clarified well. We aimed to investigate the effect of peripheral chronic nesfatin-1 application on blood pressure regulation in normal and in rats exposed to restraint immobilization stress. In our study, three month-old male Wistar rats were used. Rats were divided into 4 groups as Control, Stress, Control+Nesfatin-1, Nesfatin-1+Stress. Angiotensinogen, angiotensin converting enzyme 2, angiotensin II, endothelin-1, endothelial nitric oxide synthase, aldosterone, cortisol, nesfatin-1 levels were determined in plasma samples by ELISA. Our results have shown that chronic peripheral nesfatin-1 administration increases blood pressure in normal and in rats exposed to chronic restraint stress. Effect of nesfatin-1 on circulating level of angiotensinogen, angiotensin converting enzyme 2, angiotensin II, endothelin-1, endothelial nitric oxide synthase, aldosterone and cortisol has been identified. We can conclude that elevated high blood pressure after chronic peripheral nesfatin-1 administration in rats exposed to chronic restraint stress may be related to decreased plasma level of endothelial nitric oxide synthase concentration.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 81-88.

  Vibroacustic microvibrations enhance kidney blood supply, glomerular filtration and glutathione peroxidase activity in spontaneously hypertensive rats
Zoran Miloradović 1), Nevena Mihailović-Stanojević, Đurđica Jovović, Milan Ivanov, Una Vajić, Danijela Karanović, Jelica Grujić Milanović

1)Institute for Medical Research, University of Belgrade, Serbia.

Limited numbers of studies include research of microvibration therapy in experimental models. We examined effects of chronic vibroacustic-microvibration treatment on haemodynamics and anti-oxidative defense in experimental hypertension. Study was performed on chronically treated hypertensive and normotensive Wistar rats. Mean arterial pressure (MAP), cardiac output (CO), renal blood flow (RBF), glomerular filtration and activity of anti-oxidative enzymes were determined after three weeks treatment. Vibroacustic treatment had no influence on MAP and CO, but RBF was increased in both groups of treated rats. Additionally, vibroacustic treatment enhanced diuresis and increased glomerular filtration in hypertensive rats. Glutathione peroxidase (GSH-Px) activity was elevated in both treated rat strains, but activity of superoxide dismutase was unchanged. We conclude that microvibration treatment doesn’t ameliorate hypertension but improves renal blood supply (trough diminished renal vascular resistance), glomerular filtration, diuresis, and enhances glutathione dependent anti-oxidant defense with more important beneficials in hypertensive animals.

General Physiology and Biophysics. Volume 34, 2015, No. 1: 89-94.

  Intestinal ischemia-reperfusion injury mediates expression of inflammatory cytokines in rats
Kristína Gregová 1), Štefan Čikoš, Miroslava Bilecová-Rabajdová, Peter Urban, Ján Varga, Štefan Feterik, Jarmila Veselá

1)Department of Histology and Embryology, Faculty of Medicine, Pavol Jozef Šafárik University, Šrobárova 2, 041 80, Košice, Slovak Republic.

The small intestine is an organ with very well developed immunological activity, responsible for synthesis of specific inflammatory mediators that participate in causing the systemic inflammation that can occur after ischemia-reperfusion injury. The aim of our work was to study mRNA expression and protein concentration of inflammatory cytokines IL-10 and TNFα in the jejunal wall after intestinal ischemia-reperfusion injury (IRI). Cytokine concentration levels confirmed the direct effect of IRI on the inflammation process. The results refer to the changes in balance between pro-inflammatory and anti-inflammatory mediators and indicate that the predominant disturbance of homeostasis after intestinal IRI is present after 1 hour of reperfusion

General Physiology and Biophysics. Volume 34, 2015, No. 1: 95-99.