PUNICALAGIN AND PUNICALIN SUPPRESS THE ADIPOCYTE DIFFERENTIATION THROUGH THE TRANSCRIPTION FACTORS.

BACKGROUND: Pomegranate is a rich source of many polyphenolic compounds including ellagitannins (punicalagin, punicalin and others). AIM: The effects of punicalagin and punicalin on adipogenesis were investigated in this study. MATERIALS AND METHODS: To examine the effect of punicalagin and punicalin on adipocyte differentiation, various concentrations of punicalagin and punicalin (2-10 µM) were applied to differentiated 3T3-L1 cells. Glyceraldehyde-3-phosphate dehydrogenase (GPDH) activity, Oil red O staining, intracellular triglyceride levels, and gene expressions of transcription factors (Peroxisome proliferator-activated receptor-γ (PPARγ), CCAAT-enhancer-binding proteins-α (C/EBPα), Sterol regulatory element-binding protein 1c (SREBP-1c)) and lipolysis-associated genes (hormone-sensitive lipase (HSL), Perilipin A, tumor necrosis factor-α (TNF-α)) were examined in order to investigate the effects of punicalagin and punicalin on adipocyte differentiation. RESULTS: Punicalagin and punicalin applications caused a continuous decrease in cell size and intracellular triglyceride accumulation. GPDH activity and transcription gene expressions decreased significantly in groups that were applicated punicalagin and punicalin at high concentrations. Punicalagin, but not punicalin, down-regulated the expression of HSL and perilipin A and up-regulated the expression of TNF-α in a dose-dependent manner. In conclusion, both punicalagin and punicalin were able to inhibit the adipocyte differentiation.

Opioid peptides as possible neuromodulators in the frog peripheral nerve system.

Sciatic nerves of the frog Rana ridibunda were examined for the effects of applied opioid peptide, methionine-enkephalin, synthetic enkephalin analogue, leucine-enkephalin-NH(2) and opiate antagonist, naloxone. The effect of both peptides in concentrations of 1x10(-6) and 1x10(-5)M or naloxone in 1x10(-6)M was investigated on the action potential parameters using electrophysiological techniques. The isolated nerves were stimulated by single square pulses each of which lasted for 0.5ms at supramaximal strength. Effect of each single dose of peptides at 0min was compared with the remaining time segments. Both peptides produced changes in action potential of nerve when compared with untreated nerves. Methionine-enkephalin in both concentrations reduced the amplitude between 7% and 41% and conduction velocity at about 26-61%. This peptide in the same concentrations prolonged the duration around 12-53% and increased the stimulating voltage at about 9-50%. In contrast, leucine-enkephalin-NH(2) in both concentrations caused a decrease in amplitude between 13% and 48% and in conduction velocity around 20-50%. The same concentrations of this peptide prolonged the duration at about 3-33% and increased the stimulating voltage at about 10-56%, but naloxone in 1x10(-6)M antagonized the responses of both peptides over 75%. The results indicate that both opioid peptides produce changes in action potential parameters in frog peripheral nerve system and these changes are partially reversed by naloxone.

Evidence for the involvement of an opioid system in sciatic nerve of Rana ridibunda.

The effect of opioid peptide, D-alanine2-leucine-enkephalin and opioid homolog peptide, des-tyrosine-methionine-enkephalin in concentrations of 1 x 10(-6) and 1 x 10(-5) M was investigated on the action potential parameters of frog sciatic nerve. Des-tyrosine-methionine-enkephalin was used as the control to prove the opioid action of the peptide. The effects of both peptides were examined by means of the extracellular electrophysiological technique. The isolated sciatic nerves were stimulated by single square pulses each of which lasted for 0.5 ms at supramaximal strength. Effect of each single dose of peptides at 0 min was compared with the remaining time segments. Both peptides produced changes on action potential of Rana ridibunda sciatic nerve when compared with untreated nerves. D-alanine2-leucine-enkephalin decreased significantly the amplitude at about 34-83%, the area at about 34-92%. The same concentrations of this peptide decreased significantly the conduction velocity around 35-78%. In contrast, des-tyrosine-methionine-enkephalin reduced the action potential amplitude between 8% and 80%. The same concentrations of this peptide decreased significantly the area at about 12-76% and the conduction velocity around 42-70%. The depression of both peptides in action potential parameters was partially blocked by 1 x 10(-6) M naloxone.

Evaluation of cerebral maturation by visual and quantitative analysis of resting electroencephalography in children with primary nocturnal enuresis.

This study was undertaken to evaluate resting electroencephalographic (EEG) changes and their relations to cerebral maturation in children with primary nocturnal enuresis. Cerebral maturation is known to be important in the pathogenesis of this disorder. Twenty-five right-handed patients with primary nocturnal enuresis, aged 6 to 14 years, and 23 age- and sex-matched healthy children were included in this cross-sectional case-control study. The abnormalities detected using such techniques as hemispheral asymmetry, regional differences, and hyperventilation response in addition to visual and quantitative EEG analysis were examined statistically by multivariate analysis. A decrease in alpha activity in the left (dominant hemisphere) temporal lobe and in the frontal lobes bilaterally and an increase in delta activity in the right temporal region were observed. We concluded that insufficient cerebral maturation is an important factor in the pathogenesis of primary nocturnal enuresis, and EEG, as a noninvasive and inexpensive method, could be used in evaluating cerebral maturation.

The effects of corticosteroids and vitamin A on the healing of tracheal anastomoses.

OBJECTIVE: This study investigates the deleterious effects of corticosteroids on tracheal anastomotic healing and the ability of vitamin A to reverse these effects in a rat model. METHODS: Forty-two adult Wistar rats were randomly divided into five groups. The animals underwent tracheal transection and primary anastomoses. The groups were assigned as follows: Group I, sham (N=6); Group II, control (N=6); Group III, dexamethasone, 0.1 mg/kg/day intramuscularly (N=10); Group IV, dexamethasone 0.1 mg/kg/day intramuscularly+vitamin A 10000 IU/kg/day by gavages (N=10); and Group V, vitamin A 10000 IU/kg/day by gavages for a week (N=10). After 7 days, anastomotic healing was assessed by measurement of bursting pressure, hydroxyproline content and subsequent histological grading using the modified Ehrlich/Hunt scale. RESULTS: Bursting pressures and hydroxyproline contents were as follows: Group I: 977+/-8 mmHg and 11.80+/-0.3 microg/mg (mean+/-standard error of the mean); Group II: 890+/-55 mmHg and 9.93+/-0.6 microg/mg; Group III: 555+/-26 mmHg and 11.90+/-1.3 microg/mg; Group IV: 873+/-73 mmHg and 10.24+/-2.2 microg/mg; Group V: 905+/-45 mmHg and 7.51+/-0.8 microg/mg, respectively. Bursting pressure of Group III was found to be significantly lower when compared to other groups (P<0.0001). However, statistical significance was not found among the study groups for the hydroxyproline content. Except for inflammatory cell infiltration, histological parameters including epithelial regeneration, fibroblast proliferation, collagen content, and angiogenesis demonstrated significant differences among the groups. CONCLUSION: The present study demonstrates that dexamethasone significantly impairs the healing of tracheal anastomoses in rats and postoperative administration of vitamin A appreciably reverses this inhibitory effect. Patients receiving corticosteroids may benefit from vitamin A when undergoing prolonged intubation and laryngotracheal reconstruction.

Effects of organophosphate insecticides on mechanical properties of rat aorta.

The present study was carried out to search whether organophosphate pesticides affect the mechanical properties of the thoracic aorta. Wistar female rats (aged 6-8 weeks) were assigned randomly to a control group and groups treated with either dichlorvos or chlorpyriphos for 90 days at a dose of 5 mg/kg/day. After that period, animals were killed and thoracic aorta strips in longitudinal direction were isolated. The stress, strain and elastic modulus were obtained from the strips. Our results showed that chronic administration of chlorpyriphos and dichlorvos caused downward shift of the stress-strain relations compared to the control curve. The elastic modulus-stress curve revealed distinct characteristics in the low and high stress regions. A power function was used to simulate the low stress region while a line was fit to the high stress region. Curve fitting procedure illustrated that both pesticides influenced mainly the high stress region, but they had diverse effects at the low stress region. The results also imply that chlorpyriphos and dichlorvos decrease the strength of the aorta and therefore might influence the response of the aorta to mechanical loading induced by blood pressure.

Sepsis induces early phrenic nerve neuropathy in rats.

The aim of the present study was to investigate the electrophysiology of the phrenic nerve and the diaphragm muscle during sepsis. In total, 26 rats underwent either sham laparotomy or caecal ligation and puncture (CLP). Electrophysiology was evaluated via a phrenic nerve conduction study and needle electromyography of the diaphragm, prior to CLP, 6 and 24 h post-CLP and on day 7. The histopathology of the diaphragm muscle and phrenic nerve was also examined on day 7. In the sepsis group, the phrenic nerve conduction study showed decreased amplitude of compound action potential (CMAP), and prolongation in the duration and the latency of CMAP. The diaphragmatic needle electromyography showed decreased amplitude and frequency of the motor unit action potential (MUP), and prolongation in the duration of MUP, at all time points, compared with the pre-CLP values. The electrophysiological abnormalities were consistent with axonal and demyelinating phrenic nerve neuropathy. Electrophysiological abnormalities were present at 6 h with worsening at 24 h and on day 7. Histopathological examination showed normal muscular fibres and focally slight myelin degenerations of the phrenic nerve fibres. In conclusion, sepsis induced phrenic nerve neuropathy as early as the 6th h in rats.

Acute electrophysiological effect of pulsed gallium-arsenide low-energy laser irradiation on isolated frog sciatic nerve.

We evaluated the acute electrophysiological effects of low-energy pulsed laser irradiation on isolated frog sciatic nerve measured by extracellular recording technique. A pulsed gallium-arsenide (GaAs) laser (wavelength: 904 nm, pulse duration 220 ns, peak power per pulse: 27W, spot size: 0.28 cm(2), total applied energy density: 0.005-2.5J/cm(2)) was used for the experiment. Sixty isolated nerves were divided into six groups (n=10), each of which received a different laser dose. In each group, action potentials were recorded before laser irradiation which served as the control data. The extracellular action potentials were recorded for each combination of 1, 3, 5, 7, 10, 13 and 15 minutes of irradiation time and 4, 8, 16, 32, 64 and 128 repetition frequency by using a BIOPAC MP 100 Acquisition System Version 3.5.7 (Santa Barbara, USA). Action potential amplitude, area, duration and conduction velocity were measured. Statistical evaluation was performed using repeated measures variance analysis by SPSS 9.0. There were no statistically significant differences for action potential amplitude, area and conduction velocity among the laser groups and control data (p>0.05). The study showed that low-energy GaAs irradiation at 4-128 Hz repetition frequencies administered for irradiation times of 1-15 min generates no effect on action potential amplitude, area, duration and conduction velocity in isolated frog sciatic nerve.

No effect of GA-AS (904 nm) laser irradiation on the intact skin of the injured rat sciatic nerve.

We evaluated the electrophysiological and histopathological effects of low-energy gallium arsenide (904 nm) laser irradiation on the intact skin injured rat sciatic nerve. Twenty-four male Wistar rats were divided into three groups ( n=8 each). At the level of proximal third of the femur the sciatic nerve was crushed bilaterally with an aneurysm clip (Aesculap FE 751, Tuttingen, Germany) for half a second. A gallium arsenide laser (wavelength 904 nm, pulse duration 220 ns, peak power per pulse 27 W, spot size 0.28 cm2, pulse repetition rate 16, 128 and 1000 Hz; total applied energy density 0.31, 2.48 and 19 J/cm2) was applied to the right sciatic nerve for 15 min daily at the same time on 7 consecutive days. The same procedure was performed on the left sciatic nerve of same animal, but without radiation emission, and this was accepted as control. Compound muscle action potentials were recorded from right and left sides in all three groups before surgery, just at the end of injury, at the 24th hour and on the 14th and 21st days of injury in all rats using a BIOPAC MP 100 Acquisition System Version 3.5.7 (Santa Barbara, USA). BIOPAC Acknowledge Analysis Software (ACK 100 W) was used to measure CMAP amplitude, area, proximal and distal latency, total duration and conduction velocity. Twenty-one days after injury, the rats were sacrificed. The sciatic nerves of the operated parts were harvested from the right and left sides. Histopathological evaluation was performed by light microscopy. Statistical evaluation was done using analysis of variance for two factors (right and left sides) repeated-measures (CMAP variables within groups) and the Tukey-Kramer Honestly Significant Difference test (CMAP variables between laser groups). The significance was set at p < 0.05. No statistically significant difference (p > 0.05) was found regarding the amplitude, area, duration and conduction velocity of CMAP for each applied dose (0.31, 2.48 and 19 J/cm2) on the irradiated (right) side and the control (left) side, or between irradiated groups. Twenty-one days after injury there were no qualitative differences in the morphological pattern of the regenerated nerve fibres in either irradiated (0.31, 2.48 and 19 J/cm2) or control nerves when evaluated by light microscopy. This study showed that low-energy GaAs irradiation did not have any effect on the injured rat sciatic nerve.