Losartan pretreatment reduces neurodegeneration and behavioural symptoms in 6-hydroxydopamine induced unilateral rat model of Parkinson's disease

Losartan is an angiotensin II receptor antagonist which is mainly used to treat hypertension. It has been shown that angiotensin II involves in NADPH-dependent oxidase activation. In this study, the effect of losartan in 6-hydroxydopamine (6-OHDA)-induced rat model of Parkinson's disease was investigated. The rats were daily pre-treated i.p. with losartan (90 mg/kg), for the duration of six days before the 6-OHDA injection in the left substantia nigra pars compacta (SNC), until one day afterwards. Losartan administration caused a significant decrease in the rotational and rigidity score in the lesioned rats after 2 weeks. Furthermore, the pretreatment with losartan significantly lowered the value of the markers of oxidative stress after 24 h. Moreover, losartan protected SNC dopaminergic neurons against toxicity of 6-OHDA. The results therefore suggested that losartan pretreatment attenuated the symptoms of Parkinson's disease probably by preventing 6-OHDA induced oxidative stress and neurodegeneration.     

Different susceptibility to 1-methyl-4-phenylpyridium (MPP+)-induced nigro-striatal dopaminergic cell loss between C57BL/6 and BALB/c mice is not related to the difference of monoamine oxidase-B (MAO-B)

Subcutaneous and intraperitoneal administrations of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induce selective dopaminergic (DA-ergic) neuronal death in many animal species. After passing through the blood–brain barrier (BBB), MPTP is converted to 1-methy-4-phenylpiridinium (MPP⁺) by astrocytic monoamine oxidase-B (MAO-B). MPP⁺ then induces the dopaminergic neuronal death. In mice, marked strain differences in the susceptibility to MPTP-injection have been reported. To clarify which factor(s) cause the strain differences, MPTP or MPP⁺ was intracerebroventricularly (icv) injected into adult C57BL/6 (highly susceptible to MPTP) and BALB/c (resistant to MPTP) mice. The brain tissues including the striatum and substantia nigra pars compacta (SNpc) were examined immunohistochemically using an antibody to tyrosine hydrocyrase (TH). MPP⁺-injected C57BL/6 mice showed a significant decrease in TH-immunopositive areas in the striatum at Day 3 post injection (p < 0.01), and TH-positive cells in the SNpc at Days 1 and 3 (p < 0.01), respectively, compared to saline-injected control mice. In addition, MPP⁺-injected BALB/c mice showed a significant decrease in TH-positive areas in the striatum at Days 1 and 3, and SNpc TH-positive cells in the SNpc at Day 3, respectively (p < 0.05). However, the decrease rates in the BALB/c mice were lower than that in C57BL/6 mice. MPTP-injected C57BL/6 mice, however, showed no lesions in the striatum and SNpc at Days 1 and 7 after icv injection. All the present findings indicate that factors other than MAO-B can influence the strain susceptibility between C57BL/6 and BALB/c mice after the conversion from MPTP to MPP⁺.     

Fragmin/protamine microparticles to adsorb and protect HGF and to function as local HGF carriers in vivo

The clinical efficacy of hepatocyte growth factor (HGF) in tissue repair can be greatly enhanced by high affinity, biocompatible drug carriers that maintain the bioactivity and regulate release at the target site. We produced 0.5–3.0 μm fragmin (low molecular weight heparin)/protamine microparticles (F/P MPs) as carriers for the controlled release of HGF. F/P MPs immobilized more than 3 μg of HGF per mg of MPs and gradually released the absorbed HGF into the medium with a half-release time of approximately 5 days. Compared with HGF alone, HGF-containing F/P MPs substantially enhanced the mitogenic effect of HGF on cultured human microvascular endothelial cells, by prolonging the biological half-life, and its conjugation to F/P MPs protected HGF from heat and proteolytic inactivation. F/P MPs disappeared 8 days after subcutaneous injection in mice, suggesting that they are rapidly biodegraded. Furthermore, the number of large (diameter ?200 μm or containing ?100 erythrocytes) and medium (diameter 20–200 μm or containing 10–100 erythrocytes) lumen capillaries 8 days after injection of HGF-containing F/P MPs was significantly higher than that after injection of HGF or F/P MPs alone. Furthermore, the number of small (diameter ?20 μm or containing 1–10 erythrocytes) lumen capillaries was significantly higher 4 days after injection of HGF-containing F/P MPs. This increased angiogenic activity of HGF in vivo is probably due to both sustained local release and protection against biodegradation by the F/P MPs. Thus, F/P MPs may be useful and safe HGF carriers that facilitate cell proliferation and vascularization at sites of tissue damage.     

Does CA1 dopaminergic system play a role in cholestasis induced hypothermia?

ObjectiveThere is some of evidence describing that cholestasis induces hypothermia. Meanwhile, there is paucity of comprehensive data on the mechanism(s) governing this phenomenon. The present study was undertaken to determine the effect of CA1 dopaminergica system on cholestasis induced hypothermia.MethodsMale NMRI mice weighing 25–30 g, were used. Bilateral cannulae were implanted in dorsal hippocampi (CA1) for drug microinjection. Animals were randomly divided into non-operated control, sham-operated and bile duct-ligated (BDL) groups. Cholestasis was induced by means of main bile duct ligation. Body temperatures were measured before, two and four days after BDL.ResultsData indicated that, two and four days post BDL, the body temperature decreases as compared to the sham-operated animals, which indicates hypothermia. Intra-CA1 injection of different doses of sulpiride (SUL; 0.25, 0.5 and 0.75 μg/mouse), SCH23390 (SCH; 0.125, 025 and 0.5 μg/mouse), SKF38393 (SKF; 0.25, 0.5 and 1 μg/mouse) and quinpirole (QUI; 0.25, 05 and 0.75 μg/mouse) exerted no effect on body temperature in non-operated and sham operated mice, by themselves. Moreover, intra-CA1 injection of SUL, QUI or SKF blocked, whereas, SCH tended to increase BDL induced hypothermia.ConclusionsThe present data revealed that the CA1 dopaminergic system is possibly involved in the BDL induced impairment of thermoregulation.     

Low Serum Levels of Total Rabbit-IgG Is Associated with Acute Graft-Versus-Host Disease after Unrelated Donor Hematopoietic Stem Cell Transplantation: Results from a Prospective Study

In a prospective study we determined rabbit-IgG (r-IgG) levels in serum samples before (day 0) and after (days 1 and 7) unrelated donor hematopoietic stem cell transplantation (HSCT). Most patients suffered from a hematologic malignancy. All patients received rabbit antithymocyte globulin (ATG) as part of the conditioning for 2-4 days (2 mg/kg/day). We found a good correlation (r = 0.34, r = 0.42 and r = 0.46) between the dose of ATG and serum r-IgG levels at all 3 time points. The cumulative incidence of acute graft-versus-host disease (aGVHD) grades II-IV in patients given the 4, 6, and 8 mg/kg ATG dose was 50%, 34%, and 15% (P = .04), respectively. In patients with r-IgG ≤70 μg/mL (n = 54) the cumulative incidence of grades II-IV aGVHD was 33% compared with only 6% in those with r-IgG >70 μg/mL (n = 18), P = .023. Low serum levels of r-IgG seem to be a strong predictor for aGVHD grades II-IV in patients treated with Thymoglobulin before unrelated donor HSCT.     

Dose-dependent effect of histamine on liver function markers in immunized rabbits

ObjectiveThe present study was designed to delineate the hepatotoxicological roles of histamine dose-dependently in immunized rabbits.MethodsThe cohort comprised of three groups (II, III and IV), containing 18 rabbits each, and received subcutaneous histamine 50 μg/kg, 100 μg/kg and 200 μg/kg, respectively for 10 days (b.i.d., starting from 3 days prior to immunization until 7 days after immunization). Group I (control, n = 18) received subcutaneous sterile distilled water for 10 days. They were subsequently immunized at day 3 with intravenous injection of SRBC (1 × 10⁹ cells/ml). Blood samples were collected on pre-immunization (pre-I) day 0, as well as on days 7-, 14-, 21-, 28- and 58-post-immunization (post-I). Biochemical parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and bilirubin [total bilirubin (TB), direct bilirubin (DB) and indirect bilirubin (IB)] were determined.ResultsGroups II and IV revealed a significant decrease (on day 0-pre-I) and a significant increase (on days 7-, 14-, 21-, 28- and 58-post-I) in ALT and AST levels, when compared with the corresponding values of groups I and III while group II showed a significant increase in ALT and AST levels as compared to group IV. ALP levels in groups II, III and IV showed a significant enhancement when compared with group I. Moreover, results of TB, DB and IB demonstrated increased levels in group III when compared with groups I, II and IV. The results were found statistically significant (p < 0.05).ConclusionShort-term treatment of histamine produces dose-dependent differential patterns of hepatic dysfunctions suggestive mild liver degeneration warranting further long-term studies.     

A defect of CD16-positive monocytes can occur without disease

The CD16-positive monocytes have been first described in 1988 but to date no selective defect in the number of these cells in blood has been reported. We now describe a family in which three of four siblings lack both CD16-positive monocyte subsets, i.e. the nonclassical and the intermediate monocytes. All three had CD16-positive monocytes of 2 cells/μl or less as compared to 52 ± 18 cells/μl in healthy controls.The index case was affected by recurrent pleural effusion and infections and had evidence of an auto-inflammatory condition but no mutation of any of the relevant candidate genes. The other two siblings without CD16-positive monocytes were apparently healthy. There was no defect in serum M-CSF levels and no mutation in the M-CSF and M-CSFR genes.The data indicate that the absence of CD16-positive monocytes in blood does not lead to disease.     

Neuronal expression of c-Fos after epicortical and intracortical electric stimulation of the primary visual cortex

Electrical stimulation of the primary visual cortex (V1) is an experimental approach for visual prostheses. We here compared the response to intracortical and epicortical stimulation of the primary visual cortex by using c-Fos immunoreactivity as a marker for neuronal activation.The primary visual cortex of male Sprague Dawley rats was unilaterally stimulated for four hours using bipolar electrodes placed either intracortically in layer IV (n = 26) or epicortically (n = 20). Four different current intensities with a constant pulse width of 200 μs and a constant frequency of 10 Hz were used, for intracortical stimulation with an intensity of 0 μA (sham-stimulation), 10 μA, 20 μA and 40 μA, and for epicortical stimulation 0 μA, 400 μA, 600 μA and 800 μA. Subsequently all animals underwent c-Fos immunostaining and c-Fos expression was assessed in layer I–VI of the primary visual cortex within 200 μm and 400 μm distance to the stimulation site. C-Fos expression was higher after intracortical stimulation compared to epicortical stimulation, even though ten times lower current intensities were applied. Furthermore intracortical stimulation resulted in more focal neuronal activation than epicortical stimulation. C-Fos expression was highest after intracortical stimulation with 20 μA compared to all other intensities. Epicortical stimulation showed a linear increase of c-Fos expression with the highest expression at 800 μA. Sham stimulation showed similar expression of c-Fos in both hemispheres. The contralateral hemisphere was not affected by intracortical or epicortical stimulation of either intensities. In summary, intracortical stimulation resulted in more focal neuronal activation with less current than epicortical stimulation. This model may be used as a simple but reliable model to evaluate electrodes for microstimulation of the primary visual cortex before testing in more complex settings.     

Oxygen tolerance and occurrence of superoxide dismutase as an antioxidant enzyme in Metopus es

The free-living anaerobic ciliate Metopus es was found to possess moderate tolerance to oxygen. Direct oxygen exposure led to the death of >80% of the population within 24 h, but the remaining cells exhibited some oxygen tolerance and survived up to 4 days without any growth. Survival of the ciliate was observed only in an oxygen tension up to 7.0 μM, and higher O₂ concentrations (>7.0 μM) were found to be detrimental with a K_m value of 3.5 μM. The percentage of survival (50%) was higher when the culture was exposed to a low oxygen level (1.3 μM) and it decreased with increasing oxygen tension. No catalase activity was detected in the extract of surviving ciliates. Maximum superoxide dismutase (SOD) activity of 1.52 ± 0.4 U/mg protein was observed at 1.3 μM oxygen. SOD activity was not affected by cyanide or hydrogen peroxide, indicating that it belongs to the Mn type of SOD. Methanogenic endosymbionts in M. es lost their autofluorescence on oxygen exposure of >5.0 μM, but their viability was not permanently affected, as indicated by the maintenance of a similar number of methanogens/cell upon restoring the anaerobic condition.     

Enhanced bone regeneration in rat calvarial defects implanted with surface-modified and BMP-loaded bioactive glass (13-93) scaffolds

The repair of large bone defects, such as segmental defects in the long bones of the limbs, is a challenging clinical problem. Our recent work has shown the ability to create porous scaffolds of silicate 13-93 bioactive glass by robocasting which have compressive strengths comparable to human cortical bone. The objective of this study was to evaluate the capacity of those strong porous scaffolds with a grid-like microstructure (porosity = 50%; filament width = 330 μm; pore width = 300 μm) to regenerate bone in a rat calvarial defect model. Six weeks post-implantation, the amount of new bone formed within the implants was evaluated using histomorphometric analysis. The amount of new bone formed in implants composed of the as-fabricated scaffolds was 32% of the available pore space (area). Pretreating the as-fabricated scaffolds in an aqueous phosphate solution for 1, 3 and 6 days to convert a surface layer to hydroxyapatite prior to implantation enhanced new bone formation to 46%, 57% and 45%, respectively. New bone formation in scaffolds pretreated for 1, 3 and 6 days and loaded with bone morphogenetic protein-2 (BMP-2) (1 μg per defect) was 65%, 61% and 64%, respectively. The results show that converting a surface layer of the glass to hydroxyapatite or loading the surface-treated scaffolds with BMP-2 can significantly improve the capacity of 13-93 bioactive glass scaffolds to regenerate bone in an osseous defect. Based on their mechanical properties evaluated previously and their capacity to regenerate bone found in this study, these 13-93 bioactive glass scaffolds, pretreated or loaded with BMP-2, are promising in structural bone repair.     

Three-dimensional structural niches engineered via two-photon laser polymerization promote stem cell homing

A strategy to modulate the behavior of stem cells in culture is to mimic structural aspects of the native cell/extracellular matrix interaction. We applied femtosecond laser two-photon polymerization (2PP) to fabricate three-dimensional (3-D) microscaffolds, or “niches”, using a hybrid organic–inorganic photoresist called SZ2080. The niches, of sizes fitting in a volume of 100 × 100 × 100 μm³, were made by an external containment grid of horizontal parallel elements and by an internal 3-D lattice. We developed two niche heights, 20 and 80-100 μm, and four lattice pore dimensions (10, 20, 30 μm and graded). We used primary rat mesenchymal stem cells (MSCs) to study cell viability, migration and proliferation in the niches, up to 6 culture days. MSCs preferentially stayed on/in the structures once they ran into them through random migration from the surrounding flat surface, invaded those with a lattice pore dimension greater than 10 μm, and adhered to the internal lattice while the cell nuclei acquired a roundish morphology. In the niches, the highest MSC density was found in those areas where proliferation was observed, corresponding to the regions where the scaffold surface density available for cell adhesion was highest. The microgeometry inducing the highest cell density was 20 μm high with graded pores, in which cell invasion was favored in the central region of large porosity and cell adhesion was favored in the lateral regions of high scaffold surface density. Cell density in the niches, 17 ± 6 cells/(100 × 100 μm²), did not significantly differ from that of the flat surface colonies. This implies that MSCs spontaneously homed and established colonies within the 3-D niches. This study brings to light the crucial role played by the niche 3-D geometry on MSC colonization in culture, with potential implications for the design of biomaterial scaffolds for synthetic niche engineering.     

Multiple types of calcium channels mediate transmitter release during functional recovery of botulinum toxin type A-poisoned mouse motor nerve terminals

The involvement of different types of voltage-dependent calcium channels in nerve-evoked release of neurotransmitter was studied during recovery from neuromuscular paralysis produced by botulinum toxin type A intoxication. For this purpose, a single subcutaneous injection of botulinum toxin (1 IU; dl₅₀) on to the surface of the mouse levator auris longus muscle was performed. The muscles were removed at several time-points after injection (i.e. at one, two, three, four, five, six and 12 weeks). Using electrophysiological techniques, we studied the effect of different types of calcium channel blockers (nitrendipine, ω-conotoxin-GVIA and ω-agatoxin-IVA) on the quantal content of synaptic transmission elicited by nerve stimulation. Morphological analysis using the conventional silver impregnation technique was also made. During the first four weeks after intoxication, sprouts were found at 80% of motor nerve terminals, while at 12 weeks their number was decreased and the nerve terminals were enlarged. The L-type channel blocker nitrendipine (1 μM) inhibited neurotransmitter release by 80% and 30% at two and five weeks, respectively, while no effects were found at later times. The N-type channel blocker ω-conotoxin-GVIA (1 μM) inhibited neurotransmitter release by 50–70% in muscles studied at two to six weeks, respectively, and had no effect 12 weeks after intoxication. The P-type channel blocker ω-agatoxin-IVA (100 nM) strongly reduced nerve-evoked transmitter release (>90%) at all the time-points studied. Identified motor nerve terminals were also sensitive to both nitrendipine and ω-conotoxin-GVIA.This study shows that multiple voltage-dependent calcium channels were coupled to release during the period of sprouting and consolidation, suggesting that they may be involved in the nerve ending functional recovery process.     

The restorative effect of intramuscular injection of tetanus toxin C-fragment in hemiparkinsonian rats

The C-terminal domain of the heavy chain of tetanus toxin (Hc-TeTx) is a peptide that has a neuroprotective action against dopaminergic damage by MPP⁺, both in vitro and in vivo. The trophic effects of Hc-TeTx have been related to its ability to activate the pathways of the tropomyosin receptor kinase, which are crucial for survival process. Our group had previously shown neuroprotective effect of intramuscular Hc-TeTx treatment on animals with a dopaminergic lesion; however, there is no evidence indicating its restorative effects on advanced dopaminergic neurodegeneration. The aim of our study was to examine the restorative effects of an intramuscular injection of the Hc-TeTx fragment on the nigrostriatal system of hemiparkinsonian rats. The animals were administered with a vehicle or Hc-TeTx (20 μg/kg) in the gastrocnemius muscle for three consecutive days post-dopaminergic lesion, which was made using 6-hydroxydopamine. Post-Hc-TeTx treatment, the hemiparkinsonian rats showed constant motor asymmetry. Moreover, the ipsilateral striatum of the post-Hc-TeTx group had a lower number of argyrophilic structures and a major immunorreactivity to Tyrosine Hydroxylase in the striatum and the substantia nigra pars compacta compared to the 6-OHDA group. Our results show the restorative effect of the Hc-TeTx fragment during the dopaminergic neurodegeneration caused by 6-OHDA.     

Biodegradable gelatin microspheres enhance the neuroprotective potency of osteopontin via quick and sustained release in the post-ischemic brain

Gelatin microspheres (GMSs) are widely used as drug carriers owing to their excellent biocompatibilities and toxicologically safe degradation products. The drug release profile is easily tailored by controlling the cross-linking density and surface-to-volume ratio, i.e. size, of the GMS. In this study, we employed GMSs which are 25 μm in diameter and cross-linked with 0.03125% glutaraldehyde, to enable rapid initial and a subsequent sustained release. Therapeutic potency of human recombinant osteopontin (rhOPN) with or without encapsulation into GMSs was investigated after administrating them to rat stroke model (Sprague–Dawley; middle cerebral artery occlusion, MCAO). The administration of rhOPN/GMS (100 ng/100 μg) at 1 h post-MCAO reduced the mean infarct volume by 81.8% of that of the untreated MCAO control and extended the therapeutic window at least to 12 h post-MCAO, demonstrating a markedly enhanced therapeutic potency for the use of OPN in the post-ischemic brain. Scanning electron microscopy micrographs revealed that GMSs maintained the three-dimensional shape for more than 5 days in normal brain but were degraded rapidly in the post-ischemic brain, presumably due to high levels of gelatinase induction. After encapsulation with GMS, the duration of OPN release was markedly extended; from the period of 2 days to 5 days in normal brain, and from 2 days to 4 days in the post-ischemic brain; these encompass the critical period for recovery processes, such as vascularization, and controlling inflammation. Together, these results indicate that GMS-mediated drug delivery has huge potential when it was used in the hyperacute period in the post-ischemic brain.     

Comparison of the protective effects of various antiulcer agents alone or in combination on indomethacin-induced gastric ulcers in rats

The aim of this study which was structured with the objective of determination of the optimum protective therapy against the long term NSAID therapy-induced ulcers was to compare the gastro-protective effects of various antiulcer drugs (ranitidine, omeprazole, bismuth and misoprostol) alone or in combination with each other in different doses on indomethacin-induced gastric ulcers in rats.In this experimental study the protective effect of misoprostol (100 μg/kg/day and 10 μg/kg/day i.g.), omeprazole (5 mg/kg/day and 1.5 mg/kg/day i.p.), ranitidine (40 mg/kg/day and 10 mg/kg/day i.p.), bismuth (70 mg/kg/day and 15 mg/kg/day i.g.), combinations of misoprostol (10 μg/kg/day i.g.) plus omeprazole (1.5 mg/kg/day i.p.) and misoprostol (10 μg/kg/day i.g.) plus ranitidine (10 mg/kg/day i.p.) are investigated on indomethacin (50 mg/kg/day s.c.) induced gastric ulcers. Half an hour before indomethacin administration, each group received the above treatment regimens for 5 days. After 5-day treatment, the rats were sacrificed and histopathological and hematological examinations were performed. The following regimens were found to be effective in the prevention of indomethacin-induced gastric lesions: 100 μg/kg misoprostol, 10 μg/kg misoprostol, 5 mg/kg omeprazole, combination of 10 μg/kg misoprostol plus 1.5 mg/kg omeprazole and 10 μg/kg misoprostol plus 10 mg/kg ranitidine. The prevention rates achieved by these treatments were 71.4%, 50%, 47.6%, 52.4% and 50%, respectively. As a result of this study, misoprostol and omeprazol were found to be effective in protection against NSAID-induced gastric problems; while, ranitidine and bismuth were not. Also, the combinations of these agents were not found to have additive or synergistic effects.     

Short-term increase of serum troponin I and serum heart-type fatty acid-binding protein (H-FABP) in dogs following administration of formoterol

In this paper, changes in serum levels of the cardiac biomarkers troponin I and the heart-type fatty acid-binding protein (H-FABP) following administration of a long-acting β₂-sympathicomimeticum (long-acting beta-agonist, LABA) to dogs were measured.We measured troponin I in dogs in a 4-week repeated-dose study with inhalative administration of formoterol (13 μg/kg d) and a glucocorticoid/formoterol combination (143/16 μg/kg d). The medians of troponin I increased within 3 days in both groups, far beyond the cut-off level (0.1 μg/L), but returned to baseline levels on study day 9. The increase was more pronounced in the formoterol-only group (3.29 μg/L) compared to the glucocorticoid/formoterol combination group (1.32 μg/L).In a second study, we measured serum troponin I as well as serum H-FABP levels in several samples over 7 days in dogs, receiving a single inhalative dose of a glucocorticoid/formoterol combination (120/12 μg/kg d). The median of the troponin I concentration increased above the cut-off level within 2 h and that of H-FABP within 4 h. The medians of both parameters were temporarily above the cut-off levels even on study day 7.Both studies were conducted according to national animal welfare guidelines.To our knowledge, this is the first report that shows a corresponding increase of troponin I and H-FABP in dogs treated with formoterol. Both parameters are more sensitive in detecting a drug-induced cardiac injury compared to total LDH, total CK as well as CK MB activity. However, it is recommended to take at least three blood samples per day to assess a temporary increase of troponin I.     

Effect of low frequency electrical stimulation on spike and wave discharges of perioral somatosensory cortex in WAG/Rij rats

Low frequency electrical stimulation has been revealed that as a potential cure in patient with drug resistant to epilepsy. This study tries to evaluate the effect of low frequency electrical stimulation (LFS) on absence seizure of perioral region primary somatosensory cortex (S1po). Eighteen male WAG/Rij rats were received LFS (3 Hz, square wave, monophasic, 200 μs, and 400 μA) for 25 min into S1po for a period of five days. There is 6 animals per group .The stimulating electrodes were implanted according to stereotaxic landmarks and EEG recording was obtained 30 min before and after LFS to analyse frequency, number and duration of spike–wave discharges (SWD). The results showed that in animals with unilateral stimulating electrodes (Exp1) in first and second days and also in animals with bilateral stimulating electrodes (Exp2) in days 3rd and 4th. LFS had significant decrease effects (p < 0.05) on mean number of SWD between pre-LFS. In comparison pre-LFS to post-LFS, mean of duration in Exp2 decreased significantly. In continuous application of LFS (5 days) only the data of first day was differently significant (p < 0.05) but data of other days had no difference. Comparison of data between Exp1, Exp2 and control groups showed that the mean number of Exp1 was significantly different (p < 0.05) and mean pick frequency in Exp2 was significantly decreased in comparison with Exp1 group (p < 0.05). The LFS of S1po produces significant antiepileptic effect on absence seizure but it was not persistent till the next day and shows a short time effect.     

Expression patterns of heat shock protein 25 in carbon tetrachloride-induced rat liver injury

Heat shock protein 25 (Hsp25) is a molecular chaperone playing roles in cytoprotection. We investigated the distribution and localization of Hsp25 expression in CCl₄-induced rat hepatic lesions; liver samples were obtained from 3 h to 10 days after a single oral administration of CCl₄. Immunohistochemically, Hsp25-positive hepatocytes started to appear in the perivenular area at 6 h after CCl₄ administration. Their number and strength increased till day 1. Expression of Hsp25 mRNA significantly increased after 3 h and proceeded to increase with time till day 1. Apoptotic hepatocytes were detected around the perivenular area after 6 h. The area where Hsp25-positive hepatocytes were observed till day 1 corresponded to the area where apoptotic hepatocytes were seen. On days 2 and 3, degenerative and/or necrotic hepatocytes in the perivenular area were replaced by macrophages reacting to ED1 (for CD68) and ED2 (for CD163); Hsp25 expression was seen in hepatocytes around the perivenular area and there was a close relationship of reactive macrophages with Hsp25-positive hepatocytes, suggesting a potential role for Hsp25 in suppressing injury by inflammation. The mRNA expression of tumor necrosis factor-α, monocyte chemoattractant protein-1 and osteopontin, which can be produced by infiltrating macrophages, corresponded to that of Hsp25 from day 1 to day 3; these factors might be related to the induction of Hsp25 expression. The shift of the Hsp25 expression pattern in the liver lesion might have depended on microenvironmental conditions evoked by interactions between necrobiotic hepatocytes and infiltrating macrophages. Thus, Hsp25 expression analyses should be beneficial for evaluations of hepatotoxicants.